Development of International Learning Outcomes for Shelter Medicine in Veterinary Education: A Delphi Approach.
Shelter medicine is a veterinary discipline of growing importance. Formally accepted as a clinical specialty in the US in 2014, the practice of shelter medicine worldwide is expanding. As a topic in veterinary pre-registration (undergraduate) education, it is frequently used as an opportunity to teach primary care skills, but increasingly recognized as a subject worthy of teaching in its own right. The aim of this study was to use a Delphi consensus methodology to identify learning outcomes relevant to shelter medicine education. Shelter medicine educators worldwide in a variety of settings, including universities, non-governmental organizations and shelters were invited to participate. Participants were initially invited to share shelter medicine teaching materials. These were synthesized and formatted into Learning Outcomes (LOs) based on Bloom's taxonomy and organized into five subject-specific domains. Participants were then asked to develop and evaluate the identified LOs in two rounds of online surveys. Consensus was determined at > 80% of panelists selecting "agree" or "strongly agree" in response to the statement "please indicate whether you would advise that it should be included in a shelter medicine education program" for each LO. In the second survey, where re-wording of accepted LOs was suggested, preference was determined at > 50% agreement. Through this method, 102 agreed LOs have been identified and refined. These LOs, as well as those which did not reach consensus, are presented here. These are intended for use by shelter medicine educators worldwide, to enable and encourage the further development of this important veterinary discipline.


Detection of infections with hepatitis B virus, hepatitis C virus, and human immunodeficiency virus by analyses of dried blood spots--performance characteristics of the ARCHITECT system and two commercial assays for nucleic acid amplification.
Nowadays, dried blood spots (DBS) are primarily used to obtain diagnostic access to risk collectives such as intravenous drug users, who are prone to infections with hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV). Before DBS analyses can be used in this diagnostic context, however, a comprehensive evaluation of its performance characteristics must be conducted. To the best of our knowledge, the current study presents for the first time such essential data for the Abbott ARCHITECT system, which is currently the worldwide leading platform in this field of infection diagnostics. The investigation comprised 1,762 paired serum/DBS samples and a total of 3,524 determinations with the Abbott ARCHITECT HBsAg, anti-HBc, anti-HBs, anti-HCV and HIV-1-p24-antigen/anti-HIV 1/2 assays as well as with the artus HBV LC PCR and VERSANT HCV RNA qualitative (TMA) tests. In the context of DBS testing, a specificity of 100% was recorded for the seven serological and molecular biological assays. The analytical sensitivity of HBsAg, anti-HBc, anti-HBs, anti-HCV, HIV-1-p24-antigen/anti-HIV 1/2, HBV DNA, and HCV RNA detections in DBS eluates was 98.6%, 97.1%, 97.5%, 97.8%, 100%, 93%, and 100%, respectively. The results obtained indicate that it is today possible to reliably detect HBsAg, anti-HBc, anti-HBs, anti-HCV and HIV-1-p24 antigen/anti-HIV 1/2 with state-of-the-art analytical systems such as the Abbott ARCHITECT in DBS eluates even when a comparatively high elution volume of 1,000 μl is used. They also provide evidence for the inherent analytical limits of DBS testing, which primarily concern the anti-HBc/anti-HBs system for individuals with HIV infections and nucleic acid tests with relatively low analytical sensitivity.


The neural correlates of planning and executing actual tool use.
Human tool use is complex, and underlying neural mechanisms seem to be widely distributed across several brain systems; however, neuroimaging studies of actual tool use are rare because of experimental challenges hindering detailed analysis within one acting subject. We developed a "Tool-Carousel" that enabled us to test actual manipulation of different objects during fMRI and investigate the planning and execution of goal-directed actions. Particularly, we focused on the effects of three factors on object manipulations: the type of object manipulated, the type of manipulation, and the hand to be used. The main focus lay on the question of how complex object use compared with unspecific actions are processed and especially how such representations interact with the knowledge about the object in the action-related dorsal stream. We found that object manipulations with both right and left hand recruit a common network strongly lateralized to the left hemisphere especially during planning but also action execution. Specifically, while activity in the ventral stream was involved in processing semantic information and object properties, a dorso-dorsal pathway (i.e., superior occipital gyrus, superior parietal lobule, and dorsal premotor area) was relevant for monitoring the online control of objects and also a ventro-dorsal pathway (i.e., middle occipital gyrus, inferior parietal lobule, and ventral premotor area) was specifically involved in processing known object manipulations, such as tool use. Data further indicate an interaction of ventral stream areas, such as middle temporal gyrus and lateral occipital complex, with both dorsal pathways. These results provide evidence for left-lateralized occipito-temporo-parieto-frontal network of everyday tool use, which may help to characterize specific deficits in patients suffering from apraxia.


Refining the chemotherapy approach for older patients with colon cancer.
Population studies support an increased incidence of most cancers among older adults. Colorectal cancer has high prevalence in the aging population, with a median age of 69 years at diagnosis and 74 years at death. The vast majority of patients with colon cancer (CC) will require chemotherapy treatments during their disease course, challenging oncologists with the task of tailoring therapy for older patients with CC in the face of limited evidence-based data to guide them. Factors such as comorbidity, performance status, cognitive function, and social support may affect decision making and complicate tolerance of any recommended therapy. In recent years, attention to the specific needs of the aging population with cancer has given rise to the field of geriatric oncology in general, and has generated an increasing fund of knowledge on which to base chemotherapy delivery for this specific population of patients with CC. This article will review the available data specifically for chemotherapy management of older patients with CC in the postoperative and metastatic settings.


A potassium chloride to glycine betaine osmoprotectant switch in the extreme halophile Halorhodospira halophila.
Halophiles utilize two distinct osmoprotection strategies. The accumulation of organic compatible solutes such as glycine betaine does not perturb the functioning of cytoplasmic components, but represents a large investment of energy and carbon. KCl is an energetically attractive alternative osmoprotectant, but requires genome-wide modifications to establish a highly acidic proteome. Most extreme halophiles are optimized for the use of one of these two strategies. Here we examine the extremely halophilic Proteobacterium Halorhodospira halophila and report that medium K+ concentration dramatically alters its osmoprotectant use. When grown in hypersaline media containing substantial K+ concentrations, H. halophila accumulates molar concentrations of KCl. However, at limiting K+ concentrations the organism switches to glycine betaine as its major osmoprotectant. In contrast, the closely related organism Halorhodospira halochloris is limited to using compatible solutes. H. halophila performs both de novo synthesis and uptake of glycine betaine, matching the biosynthesis and transport systems encoded in its genome. The medium K+ concentration (~10 mM) at which the KCl to glycine betaine osmoprotectant switch in H. halophila occurs is near the K+ content of the lake from which it was isolated, supporting an ecological relevance of this osmoprotectant strategy.


Computer Aided Drug Design and its Application to the Development of Potential Drugs for Neurodegenerative Disorders.
Neurodegenerative disorders (NDs) are diverse group of disorders characterized by escalating loss of neurons (structural and functional). The development of potential therapeutics for NDs presents an important challenge, as traditional treatments are inefficient and usually are unable to stop or retard the process of neurodegeneration. Computer-Aided Drug Design (CADD) has emerged as an efficient means of developing candidate drugs for the treatment of many disease types. Applications of CADD approach to drug discovery are progressing day by day. The recent tendency in drug design is to rationally design potent therapeutics with multi-targeting effects, higher efficacies, and fewer side effects, especially in terms of toxicity. A wide literature search was performed for writing this review. An updated view on different types of NDs, their effect on human population and a brief introduction to CADD, various approaches involved in this technique, ranging from structural-based to ligand-based drug design has been discussed. The successful application of CADD approaches for the treatment of neurodegenerative disorders is also included in this review. In this review, we have briefly described about CADD and its use in the development of the therapeutic drug candidates against NDs. The successful applications, limitations and future prospects of this approach have also been discussed. CADD can assist researchers studying interactions between drugs and receptors. We believe this review will be helpful for better understanding of CADD and its applications towards the discovery of new drug candidates against various fatal NDs.


[Heterochromatin and single-strand DNA breaks (a hypothesis)].
A hypothesis is put forward which explains the known heterochromatin properties: dense DNA packing, transcriptional inactivity, a tendency for aggregation (adhesiveness) and ectopic contacts. The basic assumption of the hypothesis is that the DNA molecules in heterochromatin are topologically open and contain single-strand breaks in the regions with the same or similar base sequence.


Absolute Position Coding Method for Angular Sensor-Single-Track Gray Codes.
Single-track Gray codes (STGCs) is a type of absolute position coding method for novel angular sensors, because it has single-track property over traditional Gray codes and mono-difference over linear feedback shift register codes. However, given that the coding theory of STGCs is incomplete, STGC construction is still a challenging task even though it has been defined for more than 20 years. Published coding theories and results on STGCs are about two types of STGC, namely, necklace and self-dual necklace ordering, which are collectively called as k-spaced head STGCs. To find a new code, three constraints on generating sequences are proposed to accelerate the searching algorithm, and the complete searching result of length-6 STGCs is initially obtained. Among the entire 132 length-6 STGCs, two novel types of STGCs with non-k-spaced heads are found, and the basic structures of these codes with the general length n are proposed and defined as twin-necklace and triplet-necklace ordering STGCs. Furthermore, d-plet-necklace ordering STGC, which unifies all the known STGCs by changing the value of d, is also defined. Finally, a single-track absolute encoder prototype is designed to prove that STGCs are as convenient as the traditional position coding methods.


Genetic variant rs17225178 in the ARNT2 gene is associated with Asperger Syndrome.
Autism Spectrum Conditions (ASC) are neurodevelopmental conditions characterized by difficulties in communication and social interaction, alongside unusually repetitive behaviours and narrow interests. Asperger Syndrome (AS) is one subgroup of ASC and differs from classic autism in that in AS there is no language or general cognitive delay. Genetic, epigenetic and environmental factors are implicated in ASC and genes involved in neural connectivity and neurodevelopment are good candidates for studying the susceptibility to ASC. The aryl-hydrocarbon receptor nuclear translocator 2 (ARNT2) gene encodes a transcription factor involved in neurodevelopmental processes, neuronal connectivity and cellular responses to hypoxia. A mutation in this gene has been identified in individuals with ASC and single nucleotide polymorphisms (SNPs) have been nominally associated with AS and autistic traits in previous studies. In this study, we tested 34 SNPs in ARNT2 for association with AS in 118 cases and 412 controls of Caucasian origin. P values were adjusted for multiple comparisons, and linkage disequilibrium (LD) among the SNPs analysed was calculated in our sample. Finally, SNP annotation allowed functional and structural analyses of the genetic variants in ARNT2. We tested the replicability of our result using the genome-wide association studies (GWAS) database of the Psychiatric Genomics Consortium (PGC). We report statistically significant association of rs17225178 with AS. This SNP modifies transcription factor binding sites and regions that regulate the chromatin state in neural cell lines. It is also included in a LD block in our sample, alongside other genetic variants that alter chromatin regulatory regions in neural cells. These findings demonstrate that rs17225178 in the ARNT2 gene is associated with AS and support previous studies that pointed out an involvement of this gene in the predisposition to ASC.


Embedding value-for-money in practice: A case study of a health pooled fund programme implemented in conflict-affected South Sudan.
In recent times, there has been an increasing drive to demonstrate value for money (VfM) for investments made in public health globally. However, there is paucity of information on practical insights and best practices that have helped implementing organisations to successfully embed VfM in practice for programming and evaluation. In this article, we discuss strengths and weaknesses of approaches that been used and insights on best practices to manage for, demonstrate, and compare VfM, using a health pooled fund programme implemented in conflict-affected South Sudan as case study supported by evidence reported in the literature while critiquing adequacy of the available approaches in this setting. An expanded and iterative process framework to guide VfM embedding for health programming and evaluation is then proposed. In doing so, this article provides a very relevant one-stop source for critical insight into how to embed VfM in practice. Uptake and scale-up of the proposed framework can be essential in improving VfM and aid effectiveness which will ultimately contribute to progress towards achieving the Sustainable Development Goals by 2030.


Comparison of soluble protein kinases from normal rat prostates and prostatic Dunning tumors.
The photoaffinity labeling of the regulatory subunits of adenosine cyclic 3':5'-monophosphate-dependent soluble protein kinases with an adenosine cyclic 3':5' monophosphate analogue, 8-azidoadenosine cyclic 3':5'-monophosphate (8-N3-cAMP) was compared in normal prostates and prostatic (Dunning) tumors. Marked differences in the proportions of photoincorporation of 8-N3-[32P]cAMP into the regulatory subunit of type I (RI) and type II (RII) protein kinases and the proteolytic fragment of these regulatory subunits (Rpf) were observed in normal prostate (ventral prostate versus dorsolateral prostate) and Dunning prostatic tumors [hormone-dependent (R-3327-H) versus hormone-independent (R-3327-HI) or anaplastic (R-3327-AT) tumors]. The proportion of 8-N3-[32P]cAMP photoincorporation into RII versus RI was 2-fold higher in the dorsolateral prostate than that observed in the ventral prostate, whereas the ratio of photoincorporation into RI or Rpf was comparable between these two tissues. Comparison between tumor tissues revealed that the proportion of photoincorporation of 8-N3-[32P]cAMP into RII versus RI was 4.2-fold higher in hormone-dependent than in hormone-independent or anaplastic tumors, whereas the ratio of photoincorporation into RI versus Rpf in the former was only 6 to 7% that found in the latter. Since Dunning hormone-dependent and hormone-independent tumors have similar morphology, the present photoaffinity labeling of these regulatory subunits of soluble protein kinases provides a sensitive method (4 mg tissue per assay) to distinguish these prostatic tumors of different hormonal dependency.


In vitro studies of leukocyte labeling with technetium-99m.
A method for labeling leukocytes in vitro with 99mTc is described. Separated leukocytes are incubated with 99mTc, followed by reduction with stannous chloride and washing with acid citrate dextrose solution. Maximum labeling occurs after at least 5 min incubation with pertechnetate, followed by at least 10 min incubation with stannous chloride. Labeling is similar at room temperature and at 37 degrees C. The labeled leukocytes are viable, and reutilization of label does not occur in vitro. In acid conditions (pH 5.2), the elution of 99mTc from leukocytes is comparable with that of 32P-diisopropylfluorophosphate, but 99mTc elution is greater at pH 7.2--7.4. Neutrophils label more heavily with 99mTc than do monocytes, lymphocytes, erythrocytes, or platelets.


Unraveling Membrane Perturbations Caused by the Bacterial Riboregulator Hfq.
Hfq is a pleiotropic regulator that mediates several aspects of bacterial RNA metabolism. The protein notably regulates translation efficiency and RNA decay in Gram-negative bacteria, usually via its interaction with small regulatory RNAs. Previously, we showed that the Hfq C-terminal region forms an amyloid-like structure and that these fibrils interact with membranes. The immediate consequence of this interaction is a disruption of the membrane, but the effect on Hfq structure was unknown. To investigate details of the mechanism of interaction, the present work uses different in vitro biophysical approaches. We show that the Hfq C-terminal region influences membrane integrity and, conversely, that the membrane specifically affects the amyloid assembly. The reported effect of this bacterial master regulator on membrane integrity is discussed in light of the possible consequence on small regulatory RNA-based regulation.


Assessment of neglect reveals dissociable behavioral but not neuroanatomical subtypes.
In the current study, we investigated whether standard assessment techniques of visuospatial neglect are sensitive to detecting dissociable subtypes. We administered a battery of tasks commonly used to detect the presence of visuospatial neglect to 120 patients with unilateral right hemisphere infarcts and, in most cases, performed a systematic analysis of their lesions to quantify and localize brain damage. Using a factor analysis, we discovered seven relatively independent constructs, three of which were specifically related to the presence of left hemispatial neglect: Left Attentional Processing, Line Bisection, and Word Reading. Impairments in two of these factors, Left Attentional Processing and Line Bisection, occurred together in most cases but also occurred independently in 38 cases. There were no cases in whom Word Reading was present without concomitant deficits in one or the other two factors. These three factors could not be distinguished neuroanatomically; that is, lesions were equally likely in the temporal/parietal cortex, dorsolateral frontal cortex, or in deep frontal structures. These data confirm the notion that hemispatial neglect is a complex and multifaceted disorder composed of cognitively independent processes. These processes, however, cannot be dissociated neuroanatomically based on currently available assessment techniques.


fpocket: online tools for protein ensemble pocket detection and tracking.
Computational small-molecule binding site detection has several important applications in the biomedical field. Notable interests are the identification of cavities for structure-based drug discovery or functional annotation of structures. fpocket is a small-molecule pocket detection program, relying on the geometric alpha-sphere theory. The fpocket web server allows: (i) candidate pocket detection--fpocket; (ii) pocket tracking during molecular dynamics, in order to provide insights into pocket dynamics--mdpocket; and (iii) a transposition of mdpocket to the combined analysis of homologous structures--hpocket. These complementary online tools allow to tackle various questions related to the identification and annotation of functional and allosteric sites, transient pockets and pocket preservation within evolution of structural families. The server and documentation are freely available at http://bioserv.rpbs.univ-paris-diderot.fr/fpocket.


Suppressing lithium dendrite formation by slowing its desolvation kinetics.
Slowing the dendrite formation process is one way to alleviate the fast capacity fade and safety issues in lithium metal battery systems. We used tetraethylene glycol dimethyl ether (TEGDME) as a complementary solvent to increase the desolvation activation energy of Li+, reduce the speed of lithium electrodeposition kinetics, and suppress dendrite formation. Density functional theory calculations combined with Raman spectroscopy indicate that a stronger coordination interaction is obtained between Li+ and TEGDME than between Li+ and 1,2-dimethoxyethane (DME) or 1,3-dioxolane (DOL). Such a strong coordination leads to a slower electrochemical reaction rate. As a result, uniform lithium electrodeposition morphology and good cycling stability of a Li|Li symmetric cell for more than 500 hours were achieved. Our approach suggests a way in which dendrite formation can be controlled by the electrochemical reaction itself.


Does water hyacinth on East African lakes promote cholera outbreaks?
Cholera outbreaks continue to occur regularly in Africa. Cholera has been associated with proximity to lakes in East Africa, and Vibrio cholerae has been found experimentally to concentrate on the floating aquatic plant, water hyacinth, which is periodically widespread in East African lakes since the late 1980s. From 1994 to 2008, Nyanza Province, which is the Kenyan province bordering Lake Victoria, accounted for a larger proportion of cholera cases than expected by its population size (38.7% of cholera cases versus 15.3% of national population). Yearly water-hyacinth coverage on the Kenyan section of Lake Victoria was positively associated with the number of cholera cases reported in Nyanza Province (r = 0.83; P = 0.0010). Water hyacinth on freshwater lakes might play a role in initiating cholera outbreaks and causing sporadic disease in East Africa.


EST and EST-SSR marker resources for Iris.
Limited DNA sequence and DNA marker resources have been developed for Iris (Iridaceae), a monocot genus of 200-300 species in the Asparagales, several of which are horticulturally important. We mined an I. brevicaulis-I. fulva EST database for simple sequence repeats (SSRs) and developed ortholog-specific EST-SSR markers for genetic mapping and other genotyping applications in Iris. Here, we describe the abundance and other characteristics of SSRs identified in the transcript assembly (EST database) and the cross-species utility and polymorphisms of I. brevicaulis-I. fulva EST-SSR markers among wild collected ecotypes and horticulturally important cultivars. Collectively, 6,530 ESTs were produced from normalized leaf and root cDNA libraries of I. brevicaulis (IB72) and I. fulva (IF174), and assembled into 4,917 unigenes (1,066 contigs and 3,851 singletons). We identified 1,447 SSRs in 1,162 unigenes and developed 526 EST-SSR markers, each tracing a different unigene. Three-fourths of the EST-SSR markers (399/526) amplified alleles from IB72 and IF174 and 84% (335/399) were polymorphic between IB25 and IF174, the parents of I. brevicaulis x I. fulva mapping populations. Forty EST-SSR markers were screened for polymorphisms among 39 ecotypes or cultivars of seven species - 100% amplified alleles from wild collected ecotypes of Louisiana Iris (I.brevicaulis, I.fulva, I. nelsonii, and I. hexagona), whereas 42-52% amplified alleles from cultivars of three horticulturally important species (I. pseudacorus, I. germanica, and I. sibirica). Ecotypes and cultivars were genetically diverse - the number of alleles/locus ranged from two to 18 and mean heterozygosity was 0.76. Nearly 400 ortholog-specific EST-SSR markers were developed for comparative genetic mapping and other genotyping applications in Iris, were highly polymorphic among ecotypes and cultivars, and have broad utility for genotyping applications within the genus.


Proteomic analysis of neonatal mouse brain: evidence for hypoxia- and ischemia-induced dephosphorylation of collapsin response mediator proteins.
Perinatal hypoxia and ischemia (HI) are a significant cause of mortality and morbidity. To understand the molecular mechanisms for HI-induced brain damage, here we used a proteomic approach to analyze the alteration and modification of proteins in neonatal mouse brain 24 h after HI treatment. Significant changes of collapsin response mediator proteins (CRMPs) were observed in HI brain. CRMPs are a family of cytosolic proteins involved in axonal guidance and neuronal outgrowth. We found that CRMP2, CRMP4 and CRMP5 proteins were altered post-translationally after HI treatment. Mass spectrometric and Western blot analyses detected hypophosphorylated CRMP proteins after HI. Further analysis of CRMP kinases indicated inactivation of cyclin dependent kinase 5 (CDK5), a priming kinase of CRMPs and a neuronal specific kinase that plays pivotal roles in neuronal development and survival. The reduction of CDK5 activity was associated with underexpression of its activator p35. Taken together, our findings reveal HI-induced dephosphorylation of CRMPs in neonatal brain and suggest a novel mechanism for this modification. Hypophosphorylated CRMPs might be implicated in the pathogenesis of HI-related neurological disorders.


Peroral endoscopic myotomy versus pneumatic dilation for achalasia in patients aged ≥ 65 years.
Both peroral endoscopic myotomy (POEM) and pneumatic dilation (PD) has proved to be effective for treating achalasia in patients aged ≥ 65 years. However little is known about the comparison between POEM and PD. The aim of the study was to compare the safety and efficacy of POEM and PD for the treatment of achalasia in these patients. We retrospectively reviewed the medical records of patients aged 65 years-old or more who received POEM or PD for the treatment of achalasia at our hospital from January 2010 to December 2015, they were divided into the POEM group and the PD group. Demographics and data about safety and efficacy were collected retrospectively and compared between the two groups. A total of 31 patients were enrolled, and 21 of them received POEM, while the other 10 received PD. The treatment success (Eckardt score ≤ 3) rate of POEM and PD at 3, 6, 12, 24 and 36 months after the treatment were comparable (p > 0.05). Treatment failure was noticed in 3 cases, 1 of them was in the POEM group and the other 2 in the PD group, there was no significant difference (p > 0.05). Multivariate analysis showed that sigmoid-type achalasia was a predictive factor of treatment failure. No severe complications were observed during operation and periodical follow-up. Short-term and intermediate efficacy of POEM and PD for treating achalasia in patients aged ≥ 65 years was comparable. A large scale, randomized study with long-term follow-up is necessary in order to make a definitive conclusion.


Testing for reproductive interference in the population dynamics of two congeneric species of herbivorous mites.
When phylogenetically close, two competing species may reproductively interfere, and thereby affect their population dynamics. We tested for reproductive interference (RI) between two congeneric haplo-diploid spider mites, Tetranychus evansi and Tetranychus urticae, by investigating their interspecific mating and their population dynamics when they competed on the same plants. They are both pests of tomato, but differ in the host plant defences that they suppress or induce. To reduce the effect of plant-mediated interaction, we used a mutant tomato plant lacking jasmonate-mediated anti-herbivore defences in the competition experiment. In addition, to manipulate the effect of RI, we introduced founder females already mated with conspecific males in mild RI treatments or founder, virgin females in strong RI treatments (in either case together with heterospecific and conspecific males). As females show first-male sperm precedence, RI should occur especially in the founder generation under strong RI treatments. We found that T. urticae outcompeted T. evansi in mild, but not in strong RI treatments. Thus, T. evansi interfered reproductively with T. urticae. This result was supported by crossing experiments showing frequent interspecific copulations, strong postmating reproductive isolation and a preference of T. evansi males to mate with T. urticae (instead of conspecific) females, whereas T. urticae males preferred conspecific females. We conclude that interspecific mating comes at a cost due to asymmetric mate preferences of males. Because RI by T. evansi can improve its competitiveness to T. urticae, we propose that RI partly explains why T. evansi became invasive in Europe where T. urticae is endemic.


The Generalized Higher Criticism for Testing SNP-Set Effects in Genetic Association Studies.
It is of substantial interest to study the effects of genes, genetic pathways, and networks on the risk of complex diseases. These genetic constructs each contain multiple SNPs, which are often correlated and function jointly, and might be large in number. However, only a sparse subset of SNPs in a genetic construct is generally associated with the disease of interest. In this article, we propose the generalized higher criticism (GHC) to test for the association between an SNP set and a disease outcome. The higher criticism is a test traditionally used in high-dimensional signal detection settings when marginal test statistics are independent and the number of parameters is very large. However, these assumptions do not always hold in genetic association studies, due to linkage disequilibrium among SNPs and the finite number of SNPs in an SNP set in each genetic construct. The proposed GHC overcomes the limitations of the higher criticism by allowing for arbitrary correlation structures among the SNPs in an SNP-set, while performing accurate analytic p-value calculations for any finite number of SNPs in the SNP-set. We obtain the detection boundary of the GHC test. We compared empirically using simulations the power of the GHC method with existing SNP-set tests over a range of genetic regions with varied correlation structures and signal sparsity. We apply the proposed methods to analyze the CGEM breast cancer genome-wide association study. Supplementary materials for this article are available online.


Post-contrast myocardial T(1) and ECV disagree in a longitudinal canine study.
Both post-contrast myocardial T1 and extracellular volume (ECV) measurements have been associated with diffuse interstitial fibrosis. The cardiovascular magnetic resonance (CMR) field is migrating towards ECV, because it is largely insensitive to confounders that affect post-contrast myocardial T1 . Despite the theoretical advantages of myocardial ECV over post-contrast myocardial T1 , systematic experimental studies comparing the two measurements are largely lacking. We sought to measure the temporal changes in post-contrast myocardial T1 and ECV in an established canine model with chronic atrial fibrillation. Seventeen mongrel dogs, implanted with a pacemaker to induce chronic atrial fibrillation via rapid atrial pacing, were scanned multiple times for a total of 46 CMR scans at 3T. These dogs with different disease durations (0-22 months) were part of a separate longitudinal study aimed at studying the relationship between AF and pathophysiology. In each animal, we measured native and post-contrast T1 values and hematocrit. Temporal changes in post-contrast myocardial T1 and ECV, as well as other CMR parameters, were modeled with linear mixed effect models to account for repeated measurements over disease duration. In 17 animals, post-contrast myocardial T1 decreased significantly from 872 to 698 ms (p < 0.001), which corresponds to a 24.9% relative reduction. In contrast, ECV increased from 21.0 to 22.0% (p = 0.38), which corresponds to only a 4.5% relative increase. To partially investigate this discrepancy, we quantified collagen volume fraction (CVF) in post-mortem heart tissues of six canines sacrificed at different disease durations (0-22 months). CVF quantified by histology increased from 0.9 to 1.9% (p = 0.56), which agrees better with ECV than with post-contrast myocardial T1 . This study shows that post-contrast myocardial T1 and ECV may disagree in a longitudinal canine study. A more comprehensive study, including histologic, cardiac, and renal functional analyses, is warranted to test rigorously which CMR parameter (ECV or post-contrast myocardial T1 ) agrees better with CVF.


Myosin phenotype and bioenergetic characteristics of rat respiratory muscles.
These experiments examined the myosin phenotype and bioenergetic enzyme activities in rat respiratory muscles. Muscle samples were removed from adult female Sprague-Dawley rats (N = 8) and analyzed to determine the myosin heavy chain (MHC) and light chain (MLC) isoform content as well as the activities of myofibrillar ATPase (mATPase), citrate synthase (CS; Krebs cycle enzyme), and lactate dehydrogenase (LDH; glycolytic enzyme). Analysis revealed that CS activity and the % type I MHC and %IId MHC isoforms were greater in the costal diaphragm (CO-D) compared with those in the crural diaphragm (CR-D). In contrast, the % type IIb MHC was higher in the CR-D compared with that in the CO-D. LDH and mATPase activity were lower in both the CO-D and CR-D compared with that in the parasternal intercostals (PI), external intercostals (EI), internal intercostals (II), rectus abdominis (RA), and sternomastoid (SM) muscles. CS activity, % type I MHC, %IIa MHC, and the ratio of slow to total alkali MLC (1s/1s + 1f + 3f) were greater in the CO-D and CR-D compared with those in all other respiratory muscles. The RA contained the highest (P < 0.05) % type IIb MHC and lowest CS activity compared with that in all other muscles. Finally, CS activity, mATPase activity, and MHC phenotype did not differ among the PI, EI, II, and SM muscles. These differences in biochemical properties provide the muscles of the respiratory pump with great versatility in functional properties.


Markers for human haematopoietic stem cells: The disconnect between an identification marker and its function.
The haematopoietic system is a classical stem cell hierarchy that maintains all the blood cells in the body. Haematopoietic stem cells (HSCs) are rare, highly potent cells that reside at the apex of this hierarchy and are historically some of the most well studied stem cells in humans and laboratory models, with haematopoiesis being the original system to define functional cell types by cell surface markers. Whilst it is possible to isolate HSCs to near purity, we know very little about the functional activity of markers to purify HSCs. This review will focus on the historical efforts to purify HSCs in humans based on cell surface markers, their putative functions and recent advances in finding functional markers on HSCs.


Validation of the English and Chinese versions of the Quick-FLIC quality of life questionnaire.
A useful measure of quality of life should be easy and quick to complete. Recently, we reported the development and validation of a shortened Chinese version of the Functional Living Index-Cancer (FLIC), which we called the Quick-FLIC. In the present study of 327 English-speaking and 221 Chinese-speaking cancer patients, we validated the English version of the Quick-FLIC and further assessed the Chinese version. The 11 Quick-FLIC items were administered alongside the 11 remaining items of the full FLIC, but there appeared to be little context effect. Validity of the English version of the Quick-FLIC was attested by its strong correlation with two other measures of quality of life, and its ability to detect differences between patients with different performance status and treatment status (each P<0.001). Its internal consistency (alpha=0.86) and test-retest reliability (intraclass correlation=0.76) were also satisfactory. The measure was responsive to changes in performance status (P<0.001). The Chinese version showed similar characteristics. The Quick-FLIC behaved in ways that are highly comparable with the FLIC, even though the Quick-FLIC comprised only 11 items whereas the FLIC comprised 22. Further research is required to see whether the use of shorter instruments can improve data quality and response rates, but the fact that shorter instruments place less burden on the patients is itself inherently important.


Reassessment of non-hodgkins's lymphoma with a "nodular" growth variant: a clinicopathologic study of follicular, mantle cell and marginal zone lymphomas prospectively diagnosed with multiparameter analyses.
Although three subtypes of non-Hodgkin's lymphoma (NHL), follicular lymphoma (FL), mantle cell lymphoma (MCL) and marginal zone lymphoma (MZL), are now well recognized as independent categories, their biological behavior has not been fully compared. One of the reasons for this may be that subclassification by histological examination alone is often difficult since they all have a common variant of a "nodular" growth pattern and occasionally show similar cytological morphology. Recently, we reviewed patients with FL, MCL and MZL, who were prospectively diagnosed, using multiparameter analyses with unfixed fresh biopsy materials. Of 407 NHL patients, 101 (24.8%) belonged to these three categories and 80 could be followed; FL (n=27), MCL (n=27) and MZL (n=26). Twenty eight cases with diffuse large B-cell (DL-B) lineage lymphoma were selected as control at random. The frequency of the MCL patients with performance status (PS) 2 to 4 (41%) was significantly higher than MZL patients (4%) [P< 0.001]. The 3 year survival rate with FL, MCL, MZL and DL-B was 71.5%, 57.4%, 93.3% and 53.1%, respectively. The survival rate for MZL was significantly better than both FL (p = 0.048) and MCL (p = 0.0085). Significant differences were also found in the overall survival rates among the four risk groups as defined by the International Index [I2](low, low-intermediate, high-intermediate and high; 97.4%, 79.6%, 39.4% and 18.2%, respectively). A multivariate analysis revealed that the International Index may be a significant predictor for short survival (p=0.0001) in the patients with FL, MCL or MZL. These results suggest that MZL shows an apparently better prognosis than FL and MCL and is found to be a prognostically independent category. In contrast, the clinical outcome in MCL is the worst among the three subtypes and was closer to that of DL-B. The International Index can be applied to a wide spectrum of NHL, including MCL, MZL and FL, to and can predict prognosis in these cases.


Particulate matter 2.5 induces autophagy via inhibition of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin kinase signaling pathway in human bronchial epithelial cells.
Particulate matter 2.5 (PM2.5) is a significant risk factor for asthma. A recent study revealed that autophagy was associated with asthma pathogenesis. However, the specific mechanisms underlying PM2.5-induced autophagy in asthma have remained elusive. In the present study, PM2.5-induced autophagy was evaluated in Beas-2B human bronchial epithelial cells and the potential molecular mechanisms were investigated. Using electron microscopy, immunofluorescence staining and immunoblot studies, it was confirmed that PM2.5 induced autophagy in Beas-2B cells as a result of PM2.5-mediated inhibition of the phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway in Beas-2B cells. LY294002, a PI3K inhibitor, reduced the accumulation of microtubule-associated protein 1 light chain 3 II and attenuated the effect of PM2.5. Phosphorylated (p-)p38, p-extracellular signal-regulated kinase and p-c-Jun N-terminal kinase were dephosphorylated following exposure to PM2.5. The roles of p53, reactive oxygen species scavenger tetramethylthiourea and autophagy inhibitor 3-methyladenine in PM2.5-induced autophagy in Beas-2B cells were also investigated. The results suggested that the PI3K/Akt/mTOR signaling pathway may be a key contributor to PM2.5-induced autophagy in Beas-2B cells. The results of the present study therefore provided an a insight into potential future clinical applications targeting these signaling pathways, for the prevention and/or treatment of PM2.5-induced lung diseases.


Bullous Erythema Nodosum Leprosum as the First Manifestation of Multibacillary Leprosy: A Rare Phenomenon.
Erythema nodosum leprosum (ENL) may uncommonly present before treatment in patients with multibacillary leprosy. Atypical manifestations are known in ENL and may be clinically misleading. Such wide variations in the clinical presentation of leprosy in reaction make histopathology an important tool for supporting clinical diagnosis. We report bullous ENL presenting as the first manifestation of leprosy in a 30-year-old Indian man diagnosed using histopathology.


Cloning, expression, and sequence analysis of the Bacillus methanolicus C1 methanol dehydrogenase gene.
The gene (mdh) coding for methanol dehydrogenase (MDH) of thermotolerant, methylotroph Bacillus methanolicus C1 has been cloned and sequenced. The deduced amino acid sequence of the mdh gene exhibited similarity to those of five other alcohol dehydrogenase (type III) enzymes, which are distinct from the long-chain zinc-containing (type I) or short-chain zinc-lacking (type II) enzymes. Highly efficient expression of the mdh gene in Escherichia coli was probably driven from its own promoter sequence. After purification of MDH from E. coli, the kinetic and biochemical properties of the enzyme were investigated. The physiological effect of MDH synthesis in E. coli and the role of conserved sequence patterns in type III alcohol dehydrogenases have been analyzed and are discussed.


The D-Health Trial: A randomized trial of vitamin D for prevention of mortality and cancer.
Vitamin D, specifically serum 25(OH)D has been associated with mortality, cancer and multiple other health endpoints in observational studies, but there is a paucity of clinical trial evidence sufficient to determine the safety and effectiveness of population-wide supplementation. We have therefore launched the D-Health Trial, a randomized trial of vitamin D supplementation for prevention of mortality and cancer. Here we report the methods and describe the trial cohort. The D-Health Trial is a randomized placebo-controlled trial, with planned intervention for 5years and a further 5years of passive follow-up through linkage with health and death registers. Participants aged 65-84years were recruited from the general population of Australia. The intervention is monthly oral doses of 60,000IU of cholecalciferol or matching placebo. The primary outcome is all-cause mortality. Secondary outcomes are total cancer incidence and colorectal cancer incidence. We recruited 21,315 participants to the trial between February 2014 and May 2015. The participants in the two arms of the trial were well-balanced at baseline. Comparison with Australian population statistics shows that the trial participants were less likely to report being in fair or poor health, to be current smokers or to have diabetes than the Australian population. However, the proportion overweight or with health conditions such as arthritis and angina was similar. Observational data cannot be considered sufficient to support interventions delivered at a population level. Large-scale randomized trials such as the D-Health Trial are needed to inform public health policy and practice.


Network Analysis of MERS Coronavirus within Households, Communities, and Hospitals to Identify Most Centralized and Super-Spreading in the Arabian Peninsula, 2012 to 2016.
Contact history is crucial during an infectious disease outbreak and vital when seeking to understand and predict the spread of infectious diseases in human populations. The transmission connectivity networks of people infected with highly contagious Middle East respiratory syndrome coronavirus (MERS-CoV) in Saudi Arabia were assessed to identify super-spreading events among the infected patients between 2012 and 2016. Of the 1379 MERS cases recorded during the study period, 321 (23.3%) cases were linked to hospital infection, out of which 203 (14.7%) cases occurred among healthcare workers. There were 1113 isolated cases while the number of recorded contacts per MERS patient is between 1 (n=210) and 17 (n=1), with a mean of 0.27 (SD = 0.76). Five super-important nodes were identified based on their high number of connected contacts worthy of prioritization (at least degree of 5). The number of secondary cases in each SSE varies (range, 5-17). The eigenvector centrality was significantly (p < 0.05) associated with place of exposure, with hospitals having on average significantly higher eigenvector centrality than other places of exposure. Results suggested that being a healthcare worker has a higher eigenvector centrality score on average than being nonhealthcare workers. Pathogenic droplets are easily transmitted within a confined area of hospitals; therefore, control measures should be put in place to curtail the number of hospital visitors and movements of nonessential staff within the healthcare facility with MERS cases.


Foliar Desiccators Glyphosate, Carfentrazone, and Paraquat Affect the Technological and Chemical Properties of Cowpea Grains.
The effects of the use of glyphosate (GLY), glyphosate plus carfentrazone (GLY/CAR), and paraquat (PAR) as plant desiccators on the technological and chemical properties of cowpea grains were investigated. All studied desiccants provided lower cooking time to freshly harvested cowpea. However, the coat color of PAR- and GLY/CAR-treated cowpea was reddish in comparison to the control treatment. Principal component analysis (PCA) from liquid chromatography-mass spectrometry (LC-MS) data sets showed a clear distinction among cowpea from the different treatments. Catechin-3-glucoside and epicatechin significantly contributed for discriminating GLY-treated cowpea, while citric acid was responsible for discriminating GLY/CAR-treated cowpea. Quercetin derivative and gluconic acid were responsible for discriminating control treatment. Residual glyphosate and paraquat content was higher than the maximum limits allowed by Codex Alimentarius and the European Union Commission. Improvements in the technological and chemical properties of cowpea may not be overlapped by the risks that those desiccants exhibit when exceeding the maximum limits of tolerance in food.


Differential protective efficacy of DNA vaccines expressing secreted proteins of Mycobacterium tuberculosis.
The development of more-effective antituberculosis vaccines would assist in the control of the global problem of infection with Mycobacterium tuberculosis. One recently devised vaccination strategy is immunization with DNA plasmids encoding individual microbial genes. Using the genes for the M. tuberculosis secreted proteins MPT64 (23 kDa), Ag85B (30 kDa), and ESAT-6 (6 kDa) as candidate antigens, DNA vaccines were prepared and tested for immunogenicity and protective efficacy in a murine model of aerosolized tuberculosis (TB). Intramuscular immunization with DNA-64 or DNA-85B resulted in the activation of CD4(+) T cells, which produce gamma interferon (IFN-gamma), and high titers of specific immunoglobulin G antibodies. Further, DNA-64 induced major histocompatibility complex class I-restricted CD8(+) cytotoxic T cells. The addition of a eukaryotic leader sequence to mpt64 did not significantly increase the T-cell or antibody response. Each of the three DNA vectors stimulated a significant reduction in the level of M. tuberculosis infection in the lungs of mice challenged 4 weeks after immunization, but not to the levels resulting after immunization with Mycobacterium bovis BCG. The vaccines showed a consistent hierarchy of protection, with the most effective being Ag85B, followed by ESAT-6 and then MPT64. Coimmunization with the three vectors resulted in a greater degree of protection than that induced by any single vector. This protective efficacy was associated with the emergence of IFN-gamma-secreting T cells earlier than in infected animals immunized with a control vector. The efficacy of these DNA vaccines suggests that multisubunit vaccination may contribute to future vaccine strategies against TB.


Development and Implementation of a Family-Focused Undergraduate Nursing Curriculum: Minnesota State University, Mankato.
Nurse educators have the responsibility to create learning experiences centered on the scientific and praxis foci of the nursing discipline to advance nursing practice with families. Although the nursing profession has ample knowledge about the importance of family nursing and the value of family-focused actions, there is a lack of curricular and teaching models that address nursing practice with families in numerous courses and learning experiences. This article describes the development of a family-focused undergraduate curriculum and teaching-learning practices at Minnesota State University, Mankato in the United States. A vision and mission centered on the nursing of families, a family care teaching model, a framework of family constructs, and taxonomy of significant learning strategies guided faculty in creating learner-centered experiences. Course objectives, competencies, and teaching-learning practices in this curriculum are described. This manuscript may guide the development of innovative teaching-learning practices that integrate family nursing constructs and family nursing actions from a variety of family nursing models and theories. Initial evaluation suggests that this curriculum can increase students' knowledge of family and instill a passion for family care in undergraduate programs.


ZnO@Ag-Functionalized Paper-Based Microarray Chip for SERS Detection of Bacteria and Antibacterial and Photocatalytic Inactivation.
Bacterial infections caused by pathogenic microorganisms have become a serious, widespread health concern. Thus, it is essential and required to develop a multifunctional platform that can rapidly and accurately determine bacteria and effectively inhibit or inactivate pathogens. Herein, a microarray SERS chip was successfully synthesized using novel metal/semiconductor composites (ZnO@Ag)-ZnO nanoflowers (ZnO NFs) decorated with Ag nanoparticles (Ag NPs) arrayed on a paper-based chip as a supporting substrate for in situ monitoring and photocatalytic inactivation of pathogenic bacteria. Typical Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli and Vibrio parahemolyticus were selected as models. Partial least-squares discriminant analysis (PLS-DA) was performed to minimize the dimensionality of SERS spectra data sets and to develop a cost-effective identification model. The classification accuracy was 100, 97.2, and 100% for S. aureus, E. coli, and V. parahemolyticus, respectively. The antimicrobial activity of ZnO@Ag was proved by the microbroth dilution method, and the minimum inhibitory concentrations (MICs) of S. aureus, E. coli, and V. parahemolyticus were 40, 50, and 55 μg/mL, respectively. Meanwhile, it demonstrated remarkable photocatalytic performance under natural sunlight for the inactivation of pathogenic bacteria, and the inactivation rates for S. aureus, E. coli, and V. parahemolyticus were 100, 97.03 and 97.56%, respectively. As a result, the microarray chip not only detected the bacteria with high sensitivity but also confirmed the antibacterial and photocatalytic sterilization properties. Consequently, it offers highly prospective strategies for foodborne diseases caused by pathogenic bacteria.


The role of hydrodynamics in collective motions of fish schools and bioinspired underwater robots.
Collective behaviour defines the lives of many animal species on the Earth. Underwater swarms span several orders of magnitude in size, from coral larvae and krill to tunas and dolphins. Agent-based algorithms have modelled collective movements of animal groups by use of social forces, which approximate the behaviour of individual animals. But details of how swarming individuals interact with the fluid environment are often under-examined. How do fluid forces shape aquatic swarms? How do fish use their flow-sensing capabilities to coordinate with their schooling mates? We propose viewing underwater collective behaviour from the framework of fluid stigmergy, which considers both physical interactions and information transfer in fluid environments. Understanding the role of hydrodynamics in aquatic collectives requires multi-disciplinary efforts across fluid mechanics, biology and biomimetic robotics. To facilitate future collaborations, we synthesize key studies in these fields.


Characterization of dry globular proteins and protein fibrils by synchrotron radiation vacuum UV circular dichroism.
Circular dichroism using synchrotron radiation (SRCD) can extend the spectral range down to approximately 130 nm for dry proteins, potentially providing new structural information. Using a selection of dried model proteins, including alpha-helical, beta-sheet, and mixed-structure proteins, we observe a low-wavelength band in the range 130-160 nm, whose intensity and peak position is sensitive to the secondary structure of the protein and may also reflect changes in super-secondary structure. This band has previously been observed for peptides but not for globular proteins, and is compatible with previously published theoretical calculations related to pi-orbital transitions. We also show that drying does not lead to large changes in the secondary structure and does not induce orientational artifacts. In combination with principal component analysis, our SRCD data allow us to distinguish between two different types of protein fibrils, highlighting that bona fide fibrils formed by lysozyme are structurally more similar to the nonclassical fibrillar aggregates formed by the SerADan peptide than with the amyloid formed by alpha-synuclein. Thus, despite the lack of direct structural conclusions, a comprehensive SRCD-based database of dried protein spectra may provide a useful method to differentiate between various types of supersecondary structure and aggregated protein species.


Pseudo-precocious puberty in a Kenyan African child: a case report.
A female Kenyan child with pseudoprecocious puberty due to juvenile granulosa cell tumour is presented. Clinical features in this patient included a mass in the lower abdomen, a growth spurt and appearance of pubic hairs as well as enlargements of the breasts. No similar case has been reported in a Kenyan child. The management and review of literature are discussed.


Chronic Obstructive Pulmonary Disease In Aboriginal Patients Of The Northern Territory Of Australia: A Landscape Perspective.
The Aboriginal population of Australia has a higher burden of chronic health conditions than non-Aboriginal Australians. However, there is a paucity of data on clinical and demographic characteristics of chronic obstructive pulmonary disease (COPD) in this population. In this retrospective study we evaluated the clinical, demographic and environmental influences in adult Aboriginal patients with COPD living in the regional and remote communities of the Northern Territory of Australia. There were 380 patients (49%) with a diagnosis of COPD of the 767 patients referred to specialist respiratory outreach clinics. The mean age was 57 years (56% were female) and mean±SD BMI was 24.30±7.01 kg/m2. Smoking history was noted in 93% of the study cohort. The most common respiratory symptom was shortness of breath in 62%, and inhaled medications (salbutamol, tiotropium, salmeterol/fluticasone) were used by 79%, 44% and 58% of patients, respectively. Spirometry showed obstructive impairment (FEV1/FVC <0.7) in 79% (0.56±0.17), with mean FEV1 42% of predicted, and a bronchodilator response was present in 28.6%. Comorbid bronchiectasis was diagnosed in 49.8% along with COPD. The relationship between COPD and community demography showed a higher proportion of smokers and those with underlying bronchiectasis to have lower FEV1/FVC ratios. Communities with a higher proportion of asthma were younger and had higher smoking rates. Mortality increased with increasing number of exacerbations and hospital admissions. The Aboriginal population with COPD has a higher prevalence of smoking, moderate to severe airflow obstruction on spirometry and frequently co-diagnosed bronchiectasis with increased severity of ventilatory impairment.


Autonomic cardiac control in animal models of cardiovascular diseases. I. Methods of variability analysis.
Analysis of heart rate variability (HRV) and blood pressure variability (BPV) and baroreceptor sensitivity (BRS) has become a proven tool in clinical cardiovascular diagnostics and risk stratification. In the present work, traditional and new methodological approaches for analysis of HRV, BPV, and BRS data are summarized. HRV, BPV, and BRS parameters were obtained from animal studies designed to study pathogenetic mechanisms of distinct cardiovascular diseases. Different non-linear approaches for HRV and BPV analysis are presented here, in particular measures of complexity based on symbolic dynamics. The dual sequence method (DSM) was employed for BRS analysis. In comparison to the classical measure of BRS using the average slope [ms/mm Hg], DSM offers additional information about the time-variant coupling between BPV and HRV. Since cardiovascular regulation shares common features among different species, data on HRV and BPV, as well as BRS, in animal models might be useful for understanding the pathogenetic mechanisms of cardiovascular diseases in humans and in the development of new diagnostic approaches.


Outcomes and complications following spinous process fixation: a single-center analysis of 192 cases.
Spinous process fixation (SPF) is presented as less invasive than pedicle screws. There has been little quantitative data to support this assertion, and "minimally invasive" has not been well defined in spine surgery. Length of stay (LOS) and blood loss (BL) were chosen as surrogate measures of "minimally invasive." A chart review was conducted on 192 lumbar fusion patients (374 levels). A backward-selection multiple-linear-regression was performed to determine what variables contribute to LOS and estimated blood loss (EBL). A logistic regression controlling for age and number of levels on complication rates was also performed. Number of levels with supplementary screw fixation (SSF) was significantly associated with LOS (p = 0.003). Controlling for number of surgical levels, LOS increased by 0.30 days (95% CI: 0.02-0.58) for each level with SSF. For each additional level including SSF, BL increases by 25.31 cc (95% CI: 3.50-47.12, p = 0.023). Interbody fusion increases blood loss by 68.16 cc (95% CI: 17.18-119.13, p = .009). For each additional level with SSF, odds of perioperative complications increase by OR = 2.34 (95% CI: 1.35-4.05). Long-term complications were not affected by instrumentation. LOS and BL are increased in patients with SSF vs. SPF only. Odds of perioperative complications are increased in patients with SSF relative to those treated with SPF alone.


Role of 24-hour Helpline Service in the Management of Diabetes During the Holy Month of Ramadan.
Objective To assess the role of the 24-hour helpline service in the management of diabetes during the holy month of Ramadan. Methodology This prospective study was conducted at the Baqai Institute of Diabetology & Endocrinology, Baqai Medical University, Karachi, Pakistan, between December 2017 and August 2018. Patients with type 1 and type 2 diabetes who were willing to observe the fast in Ramadan 2018 were included. Pre-Ramadan education on a one-to-one basis was given and the 24-hour helpline number was provided to each patient. Details of baseline characteristics, anthropometric measurements, and biochemical parameters (lipid profile, serum creatinine, urine D/R (protein), and glycated hemoglobin A1c pre- and post-Ramadan) were noted. Self-monitoring blood glucose (SMBG) recording books were provided to each patient to record blood sugar levels during Ramadan. Results A total of 102 patients, 41 (40.2%) with type 1 diabetes and 61 (59.8%) with type 2 diabetes were included. The mean age of patients with type 1 diabetes was 17.02±5.22 years and that of patients with type 2 diabetes was 49.48±9.68 years. Most of the patients were male in both groups. By using the 24-hour helpline service, 912 calls were made to patients, 502 calls for type 1, and 410 calls for type 2 diabetes. Mean HbA1c (%) level before and after Ramadan was 8.94±1.72 and 8.70±2.03 in patients with type 1 diabetes and 8.59±1.73 and 7.71±1.27 in patients with type 2 diabetes, respectively. In the last 10 days of Ramadan, the decreasing trends of mean self-monitoring of blood glucose (SMBG) in patients with type 1 diabetes was observed for pre-dawn meal (suhoor), pre-sunset meal (iftar), and before sleeping while this trend in type 2 diabetes was found in the pre-dawn meal (suhoor)and two hours after iftar. Conclusion Pre-Ramadan diabetes education and treatment adjustment by using a station-based, 24-hour helpline service during the holy month of Ramadan have a significant role in controlling blood sugar levels, thus minimizing the complications of diabetes, facilitating timely adjustments in treatment, and reducing unnecessary hospital visits and admissions.


Oestrogen-induced changes in the relative concentrations of ribonuclease and ribonuclease inhibitor in rat uterus.
The cytoplasm of the immature rat uterus contains an excess of ribonuclease inhibitor over endogenous cytoplasmic ribonuclease. The ratio of free inhibitor to bound inhibitor is approximately 3:1. After treatment with oestrogen, there is a marked increase in the level of cytoplasmic ribonuclease such that the enzyme is present in excess of the inhibitor. The increase in activity appears to result from at least 2 distinct ribonucleases with molecular masses of approximately 14000 and 18000. The changes observed after oestrogen treatment are also evident during normal development of the rat uterus. The implications of these results on the proposed role of the inhibitor/ribonuclease system are discussed.


Natural variation at the FRD3 MATE transporter locus reveals cross-talk between Fe homeostasis and Zn tolerance in Arabidopsis thaliana.
Zinc (Zn) is essential for the optimal growth of plants but is toxic if present in excess, so Zn homeostasis needs to be finely tuned. Understanding Zn homeostasis mechanisms in plants will help in the development of innovative approaches for the phytoremediation of Zn-contaminated sites. In this study, Zn tolerance quantitative trait loci (QTL) were identified by analyzing differences in the Bay-0 and Shahdara accessions of Arabidopsis thaliana. Fine-scale mapping showed that a variant of the Fe homeostasis-related FERRIC REDUCTASE DEFECTIVE3 (FRD3) gene, which encodes a multidrug and toxin efflux (MATE) transporter, is responsible for reduced Zn tolerance in A. thaliana. Allelic variation in FRD3 revealed which amino acids are necessary for FRD3 function. In addition, the results of allele-specific expression assays in F1 individuals provide evidence for the existence of at least one putative metal-responsive cis-regulatory element. Our results suggest that FRD3 works as a multimer and is involved in loading Zn into xylem. Cross-homeostasis between Fe and Zn therefore appears to be important for Zn tolerance in A. thaliana with FRD3 acting as an essential regulator.


Number of potential patients to be treated with proton therapy in Italy.
In radiotherapy, the use of proton beams is one of the most promising approaches in order to reduce the treatment volume and, consequently, increase the total dose avoiding severe complications to the normal tissues surrounding the target. Among the new hospital-based facilities that are planned for the next future, the italian project is in an advanced stage of study. Because of the complexity and high cost of the italian centre, one of the most important information in order to establish the cost/benefit balance, is represented by the number of potential patients to be treated for the various pathologies. For this reason it is useful to define a priority scale of clinical indications and, on this basis, to estimate the yearly patient afflux to the center, taking into account the incidence and the expected new cases to be treated with protons. Indications have been divided into two categories, according to decreasing priority. Category A includes all the tumors in which the use of proton therapy has clearly demonstrated to be advantageous, being the only way to give a curative dose to the target. In Italy, the estimated number of this category of patients is 825 each year. Category B comprises a great variety of tumors characterized mainly by a local evolution, with a limited likelihood of distant spread, and therefore potentially cured if the local control can be obtained. The total number of potential patients in this category is more than 10,000.


Differential effects of DNA synthesis inhibitors on DNA methylation in normal and transformed cells.
1-beta-D-Arabinofuranosylcytosine (ara-C) and aphidicolin are well known inhibitors of DNA synthesis, each one acting through a different mechanism. We have examined the effects of these compounds on DNA methylation in normal human embryonic lung fibroblasts (WI-38) as well as in their Simian Virus 40 (SVWI-38) and gamma radiation (CT-1) transformed counterparts. Analysis of the methylation status of the total genomic DNA in WI-38 cells revealed that hydroxyurea was the only drug which resulted in a significant increase in the 5-methylcytosine content under conditions where greater than 98% inhibition of DNA synthesis was achieved. Under the same conditions, all three drugs were capable of inducing hypermethylation in the SVWI-38 cells, whereas none of them showed any effect on the methylation status of the DNA in the CT-1 cells. In cells where a limited degree of replication was allowed to occur at drug concentrations resulting in 50% inhibition of DNA synthesis, a different pattern emerged. Under these conditions, DNA which was synthesized in the presence of either ara-C or aphidicolin was significantly hypermethylated in both the transformed cell lines, whereas hydroxyurea had no effect. In the normal WI-38 cells however, hydroxyurea was still the only drug which caused any significant hypermethylation. Different cells thus responded differently to these three agents, and the mere slowing down of DNA synthesis did not ipso facto lead to increased DNA methylation.


Progressive massive fibrosis and simple pneumoconiosis in ex-miners.
A group of 17 738 working miners, medically examined during 1953-8, were followed up from 1974 to 1980. Of the 7118 men re-examined, 2547 were still working miners and 4526 had left the industry (45 were of unrecorded status). The incidence of progressive massive fibrosis (PMF) over an average follow up period of 22 years among men who had remained in the industry was 27 per 1000, but 94 per 1000 among men who had left. This difference was only partly related to the difference in age between the groups; for men without simple pneumoconiosis at the start of the period, and for similar age groups (45-64), the attack rate in miners was 20 per 1000 and in the ex-miners 41 per 1000. In a group of 1902 leavers who did not have PMF at a medical examination conducted at most four years before leaving, 172 had developed PMF by the time of the follow up examination. Of these, 116 had had simple pneumoconiosis at the earlier examination. Cumulative exposure to respirable dust, category of simple pneumoconiosis, and age were each found to influence the probability of developing PMF in a subgroup of the 1902 men. Among the 1902 leavers, there was no overall progression or regression of simple pneumoconiosis.


The critical roles of STING in mitochondrial homeostasis.
The stimulator of interferon genes (STING) is a crucial signaling hub in the immune system's antiviral and antimicrobial defense by detecting exogenous and endogenous DNA. The multifaceted functions of STING have been uncovered gradually during past decades, including homeostasis maintenance and overfull immunity or inflammation induction. However, the subcellular regulation of STING and mitochondria is poorly understood. The main functions of STING are outlined in this review. Moreover, we discuss how mitochondria and STING interact through multiple mechanisms, including the release of mitochondrial DNA (mtDNA), modulation of mitochondria-associated membrane (MAM) and mitochondrial dynamics, alterations in mitochondrial metabolism, regulation of reactive oxygen species (ROS) production, and mitochondria-related cell death. Finally, we discuss how STING is crucial to disease development, providing a novel perspective on its role in cellular physiology and pathology.


Serum oxidizability potential of ischemic heart disease patients is associated with exercise test results and disease severity.
To find out whether serum oxidizability potential correlates with exercise test (EXT) parameters and predicts their results in chronic ischemic heart disease (IHD) patients. Oxidizability potential was determined in a group of chronic IHD patients who underwent a symptom limited EXT upon initiation of a cardiac rehabilitation program. The thermo-chemiluminescence (TCL) assay was used to assess serum oxidizability potential. This assay is based on heat-induced oxidation of serum, leading to the formation of electronically excited species in the form of unstable carbonyls, which further decompose into stable carbonyls and light energy (low chemiluminescence). Measured photons emission is represented by a kinetic curve which is described by its amplitude and slope (= ratio). We assessed the correlations of TCL ratio with exercise duration, metabolic equivalents (METS), maximal heart rate (mHR), maximal systolic BP, > 1 mm S-T depression, diabetes, hypertension, smoking, left ventricular ejection fraction (LVEF) > or < 40%, previous myocardial infarction, and aorto-coronary by-pass surgery and compared to the TCL ratio measured in a group of healthy controls. A high TCL ratio (%) correlated well with METS (r = 0.84), with mHR (r = 0.79) and with exercise induced S-T segment shift (r = 0.87, P < 0.05). A lower serum oxidizability potential, expressed as a low TCL ratio, thus suggestive of a previous high oxidative stress, was found in IHD patients compared to healthy controls, and, in particular, in patients with low LVEF%. The TCL ratio (%) in IHD patients was 193 ± 21, compared to 215 ± 13 in controls (P < 0.05), and was 188 ± 14.7 in patients with LVEF < 40% as compared to 200 ± 11.9 in those with LVEF > 40% (P < 0.01). A trend for lower TCL ratio (%) was found in diabetic, hypertensive, and post-coronary bypass surgery patients. A paradoxically low TCL ratio (low oxidizability potential) was observed in patients without S-T depression compared to patients with S-T depression (189 ± 22 vs 201 ± 15, P = NS), due to the fact these patients had a much lower LVEF% and a lower exercise capacity. Serum oxidizability potential is associated with EXT parameters, results, and IHD severity. TCL ratio is an "easy-to-measure marker" that might be incorporated into risk assessment and prediction in chronic IHD patients.


Virtual reality videos for training and protocol dissemination during a pandemic.
This article was migrated. The article was marked as recommended. Preparations for the COVID-19 pandemic required healthcare teams to practice known skills, such as intubation, with renewed consideration for safety, as well as develop new Standard Operating Procedures (SOPs) for health care delivery. In these conditions, translational simulation based-education (SBE) is a well-known tool that supports health care teams to improve the system using design thinking methods such as walkthroughs and team-based simulation. However, the pandemic has introduced two stressors on translational SBE simultaneously. Firstly, the need for rapid upskilling of front-line staff and rapid change to SOPs. Secondly, the need for social or physical distancing at work, such that it quickly became inappropriate for large groups of individuals to practice in-situ SBE and debrief together in close proximity. An educational approach that brings the best of translational SBE while minimizing contact and maximizing experiential learning is needed. Digital learning has been rapidly adopted by much of medical education during the pandemic. Focusing on a strong alignment between learning goals with intended clinical performance change outcomes we sought to leverage a digital education format that allowed for low barriers to adoption, yet supported the experiential, dynamic reality of translational SBE. In the absence of the ability to quickly train large numbers of people due to the need for social distancing, an immersive experience that can only be provided by virtual reality (VR) videos was the next best thing. VR, using 360-degree video, supported the creation of instructional videos from SBE events in the hospital which allow the learner to immerse and explore multiple points within the scenario. We describe how the very act of recording a video assisted in the rapid development of SOPs through translational simulation. We then describe the use of VR to stay true to the spirit of simulation for experiential learning and nearly hands-on training.


Effective Information Provision About the Side Effects of Treatment for Malignant Lymphoma: Protocol of a Randomized Controlled Trial Using Video Vignettes.
Informing patients with cancer about the possible implications of prospective treatment is a crucial yet challenging task. Unfortunately, patients' recall of medical information is generally poor and their information needs are not met. Effective information giving entails that oncologists help patients understand and recall the implications of their treatment, meanwhile fostering a trusting physician-patient relationship. Communication strategies that are often suggested to be effective are structuring and tailoring (cognition-oriented) but also are oncologists' expressions of caring or empathy (affect-oriented). The aim of this study is to provide evidence concerning the pathways linking physician communication to (improved) consultation outcomes for patients. More specifically, the aim is to determine the effects of information structuring and information tailoring, combined with physician caring, on information recall, satisfaction with information, and trust in the physician (primary objective) and on symptom distress (secondary objective). A randomized controlled trial, systematically testing the effects of information structuring and information tailoring, each combined with caring, in 2 video-vignette experiments (2×2 and 2×2×2 design). Using an online survey platform, participants will be randomly allocated (blinded) to 1 of 12 conditions in which they are asked to view a video vignette (intervention) in which an oncologist discusses a treatment plan for malignant lymphoma with a patient. The independent variables of interest are systematically varied across conditions. The outcome measures are assessed in a survey, using validated instruments. Study participants are (former) patients with cancer and their relatives recruited via online panels and patient organizations. This protocol discusses the trial design, including the video-vignette design, intervention pretesting, and a pilot study. Data collection has now been completed, and preliminary analyses will be available in Spring 2019. A total of 470 participants completed the first part of the survey and were randomized to receive the intervention. The results of the proposed trial will provide evidence concerning the pathways linking physician information, giving skills to (improved) consultation outcomes for patients. Netherlands Trial Register NTR6153; https://www.trialregister.nl/trial/6022 (Archived by Webcite at http://www.webcitation.org/76xVV9xC8). DERR1-10.2196/12453.


Evolution of the selfing syndrome: Anther orientation and herkogamy together determine reproductive assurance in a self-compatible plant.
Capacity for autonomous self-fertilization provides reproductive assurance, has evolved repeatedly in the plant kingdom, and typically involves several changes in flower morphology and development (the selfing syndrome). Yet, the relative importance of different traits and trait combinations for efficient selfing and reproductive success in pollinator-poor environments is poorly known. In a series of experiments, we tested the importance of anther-stigma distance and the less studied trait anther orientation for efficiency of selfing in the perennial herb Arabis alpina. Variation in flower morphology among eight self-compatible European populations was correlated with efficiency of self-pollination and with pollen limitation in a common-garden experiment. To examine whether anther-stigma distance and anther orientation are subject to directional and/or correlational selection, and whether this is because these traits affect pollination success, we planted a segregating F2 population at two native field sites. Selection strongly favored a combination of introrse anthers and reduced anther-stigma distance at a site where pollinator activity was low, and supplemental hand-pollination demonstrated that this was largely because of their effect on securing self-pollination. The results suggest that concurrent shifts in more than one trait can be crucial for the evolution of efficient self-pollination and reproductive assurance in pollinator-poor habitats.


Predictors of quality of life in patients with skin melanoma at the dermatology department of the Porto Alegre Teaching Hospital.
Some symptoms present in melanoma patients are directly related to psychological stress, which emphasizes the need to evaluate quality of life (QoL) in these patients at all the stages of their disease. The objective of this study was to evaluate quality of life in a sample of patients diagnosed with melanoma, using the Functional Assessment of Cancer Therapy-General (FACT-G) questionnaire. A descriptive, cross sectional study was conducted between July and December, 2006 with all patients with skin melanoma receiving follow-up care at the Department of Dermatology of the Porto Alegre Teaching Hospital, Federal University of Rio Grande do Sul. Sixty patients were included in the study. Mean age was 55.6 years. Poor education level (primary school or less) was associated with a poorer FACT-G score. Patients with a family history of the disease had higher QoL scores in 3 of the 4 categories evaluated: physical, emotional and functional wellbeing (p<0.01). QoL scores were higher in married patients (82.42) compared to single patients (70.28) (p<0.01). Patients with metastases had lower scores in the functional wellbeing category and this difference was statistically significant. Factors related to the tumor, as well as gender, age and employment status, were not found to be predictive of quality of life in this sample. Quality of life scores were lower in the functional wellbeing domain in patients with metastases. Married patients are able to count on greater comfort and emotional support to help them deal with the diagnosis of melanoma. Patients with a family history of melanoma had significantly higher quality of life scores, while those with poor education levels had lower scores.


First direct evidence of chalcolithic footwear from the near eastern highlands.
In 2008, a well preserved and complete shoe was recovered at the base of a Chalcolithic pit in the cave of Areni-1, Armenia. Here, we discuss the chronology of this find, its archaeological context and its relevance to the study of the evolution of footwear. Two leather samples and one grass sample from the shoe were dated at the Oxford Radiocarbon Accelerator Unit (ORAU). A third leather sample was dated at the University of California-Irvine Accelerator Mass Spectrometry Facility (UCIAMS). The R_Combine function for the three leather samples provides a date range of 3627-3377 Cal BC (95.4% confidence interval) and the calibrated range for the straw is contemporaneous (3627-3377 Cal BC). The shoe was stuffed with loose, unfastened grass (Poaceae) without clear orientation which was more than likely used to maintain the shape of the shoe and/or prepare it for storage. The shoe is 24.5 cm long (European size 37), 7.6 to 10 cm wide, and was made from a single piece of leather that wrapped around the foot. It was worn and shaped to the wearer's right foot, particularly around the heel and hallux where the highest pressure is exerted in normal gait. The Chalcolithic shoe provides solid evidence for the use of footwear among Old World populations at least since the Chalcolithic. Other 4th millennium discoveries of shoes (Italian and Swiss Alps), and sandals (Southern Israel) indicate that more than one type of footwear existed during the 4th millennium BC, and that we should expect to discover more regional variations in the manufacturing and style of shoes where preservation conditions permit.


Depth-Dependent Corneal Biomechanical Properties in Normal and Keratoconic Subjects by Optical Coherence Elastography.
Compare depth-resolved biomechanical properties in normal and keratoconic corneas in live human subjects using optical coherence elastography (OCE). In a prospective series of normal and keratoconus (KC) eyes, a corneal perturbation was applied by a custom swept-source OCE system using a transparent flat lens coupled to force transducers. Cross-correlation was applied to track frame-by-frame OCT speckle displacement. Regional displacements for the anterior and posterior stroma were plotted in force versus displacement (k) graphs. A spatial biomechanical property ratio (ka/kp ) was defined by dividing the maximum total displacement by the maximum force for the anterior (ka ) and posterior cornea (kp) and was compared between normal and KC groups with the Mann-Whitney U test. Area under the receiver operating characteristics curve (AUROC) for differentiating normal and KC eyes was calculated for ka/kp , kmax, and thinnest point of corneal thickness (TPCT). Thirty-six eyes were analyzed (21 eyes of 12 normal subjects and 15 KC eyes of 12 subjects). The ka/kp for the normal group was 1.135 ± 0.07 (mean ± standard deviation) and 1.02 ± 0.08 for the KC group (P < 0.001), indicating a relative deficit in anterior stromal stiffness in KC eyes. AUROC was 0.91 for ka /kp , 0.95 for kmax, and 1 for TPCT. Significant differences in depth-dependent corneal biomechanical properties were observed between normal and KC subjects. OCE was applied for the first time to human KC subjects and revealed alterations in the normal anterior-to-posterior stromal stiffness gradient, a novel and clinically accessible disease biomarker.


Bioactive glycosides from the African medicinal plant Boerhavia erecta L.
Phytochemical studies of the previously unexplored stem of Boerhavia erecta from Burkina Faso, resulted in the isolation of an unreported glycoside 4, 2,3-dihydroxypropylbenzoate-3-O-β-[4″-methoxy] glucuronide as well as seven known glycosides (1-3, 5-8). The major isolate 5 and 8 indicated a significant inhibition against HIV integrase (IC50 10 and 22 μg/mL, respectively). The extracts and isolates were also tested for anti-malarial activity, but insignificant activity was observed.


Effects of Experienced Discrimination in Pediatric Sickle Cell Disease: Caregiver and Provider Perspectives.
For Black children with sickle cell disease (SCD) and their families, high disease stigmatization and pervasive racism increase susceptibility to discrimination in healthcare settings. Childhood experiences of discrimination can result in medical nonadherence, mistrust of healthcare providers, and poorer health outcomes across the lifespan. Caregivers and medical providers are essential to childhood SCD management and are therefore well-positioned to provide insight into discrimination in the context of pediatric SCD. This mixed-methods study sought caregivers' and providers' perspectives on processes underlying discrimination and potential solutions to mitigate the negative effects of perceived discrimination among children with SCD. Caregivers (N = 27) of children with SCD (≤ 12 years old) and providers from their hematology clinics (N = 11) participated in individual semi-structured interviews exploring experiences of discrimination and daily SCD management and completed a quantitative measure of discrimination. Qualitative data were collected until themes reached saturation and subsequently transcribed verbatim, coded, and analyzed using applied thematic analysis. Quantitative and qualitative data converged to suggest the pervasiveness of discrimination in healthcare settings. Three qualitative themes emerged: (1) healthcare system factors underlie discrimination, (2) families' challenging interactions with providers lead to perceptions of discrimination, and (3) experiences of discrimination impact caregiver-provider interactions. Both caregivers and providers highlighted building trusting patient-provider relationships and encouraging patients' self-advocacy as means to reduce experiences and impacts of discrimination. These findings offer potential approaches to tangibly mitigate occurrences of discrimination in pediatric healthcare settings by trust building, accountability keeping, and fostering rapport to improve quality of care and pediatric SCD health outcomes.


The PIG-anchoring defect in NK lymphocytes of PNH patients.
Paroxysmal nocturnal hemoglobinuria (PNH) is clinically characterized by intravascular hemolysis, hemoglobinuria, iron deficiency anemia, and venous thrombosis. Pathophysiologically the disease has now been generally accepted as an acquired defect of phosphatidylinositol glycan (PIG)-anchored molecules on the cell surface of bone marrow-derived cells. This defect is functionally characterized by an abnormal susceptibility to complement-mediated lysis and has been described on erythrocytes, granulocytes, monocytes, and platelets. In contrast, contradictory data exist so far on the involvement of lymphocytes and natural killer (NK) cells. Using monoclonal antibodies (MoAbs) against newly defined PIG-linked surface structures such as CD48, CD55, and CD59, which are homogeneously expressed on lymphocytes of normal donors, we analyzed lymphocytes and their subpopulations in nine PNH patients by two color immunofluorescence. Our results showed that CD3+ T cells as well as CD16+ NK cells are at least partially involved in the deficient PIG-molecule surface expression. To more clearly define the defect in PNH, we generated NK clones from a PNH patient. Phenotypic analysis of these NK clones showed that they either were positive (n = 3) for PIG-linked surface structures such as CD48, CD55, and CD59 (eg, NKP1) or were completely negative (n = 7) for all of them (eg, NKP1). In functional tests the PIG-molecule negative clone NKP2 showed increased susceptibility to human complement compared with the PIG molecule positive clone NKP1. When analyzing the mRNA levels of the PIG-linked molecules CD55 and CD59 there was no difference at all between the two clones. We conclude from our data that NK cells as well as other lymphocyte subpopulations are involved in the PIG-linkage defect of PNH. These NK clones with differential expression of PIG-linked surface structures present for the first time ex vivo mutant cell lymphocyte lines that carry the defect leading to PIG deficiency in PNH.


Application of tryptamine as a derivatizing agent for the determination of airborne isocyanates. Part 7. Selection of impinger solvents and the evaluation against dimethyl sulfoxide used in US NIOSH Regulatory Method 5522.
The application of tryptamine to derivatize airborne isocyanates has been evaluated and adopted by the National Institute for Occupational Safety and Health (NIOSH) as the latest isocyanates regulatory method (Method 5522) in the USA. Method 5522 uses dimethyl sulfoxide (DMSO) as the impinger solvent in which tryptamine is dissolved to sample isocyanates for analysis. Since DMSO is both extremely hygroscopic and corrosive, it is not a satisfactory solvent for impinger air sampling of isocyanates. The high boiling and freezing points also present some distinct drawbacks. In a search for a suitable impinger solvent, the efficiencies of various solvents, which all were potentially more suitable for sampling isocyanates viz., N,N'-dimethylformamide, butyl acetate, isobutyl acetate, sec-butyl acetate, tert-butyl acetate and octane, were investigated. Simulated air sampling was conducted on two commonly used isocyanates in industry, hexamethylene diisocyanate (HDI) and toluene diisocyanate (TDI), which were vaporized and sampled into impingers containing dissolved tryptamine. Butyl acetate and octane were found to be most suitable for using as impinger solvents. Recoveries of isocyanates at two concentration levels of HDI and TDI were 93.4-108% in comparison with those of using DMSO.


[The pattern electroretinogram in diabetic retinopathy].
The pattern electroretinogram (PERG) was examined in 79 eyes (47 patients) of diabetic retinopathy (DR), with the results that the amplitude of b-wave was significantly reduced early in the subclinical phase of DR, and the latency was lengthened from stage I on. The value of PERG in the early functional diagnosis of DR was discussed.


Real-time PCR assays for quantification and differentiation of Vibrio vulnificus strains in oysters and water.
Vibrio vulnificus is an autochthonous estuarine bacterium and a pathogen that is frequently transmitted via raw shellfish. Septicemia can occur within 24 h; however, isolation and confirmation from water and oysters require days. Real-time PCR assays were developed to detect and differentiate two 16S rRNA variants, types A and B, which were previously associated with environmental sources and clinical fatalities, respectively. Both assays could detect 10(2) to 10(3) V. vulnificus total cells in seeded estuarine water and in oyster homogenates. PCR assays on 11 reference V. vulnificus strains and 22 nontarget species gave expected results (type A or B for V. vulnificus and negative for nontarget species). The relationship between cell number and cycle threshold for the assays was linear (R(2) = >0.93). The type A/B ratio of Florida clinical isolates was compared to that of isolates from oysters harvested in Florida waters. This ratio was 19:17 in clinical isolates and 5:8 (n = 26) in oysters harvested from restricted sites with poor water quality but was 10:1 (n = 22) in oysters from permitted sites with good water quality. A substantial percentage of isolates from oysters (19.4%) were type AB (both primer sets amplified), but no isolates from overlying waters were type AB. The real-time PCR assays were sensitive, specific, and quantitative in water samples and could also differentiate the strains in oysters without requiring isolation of V. vulnificus and may therefore be useful for rapid detection of the pathogen in shellfish and water, as well as further investigation of its population dynamics.


An exploratory prospective study of marijuana use and mortality following acute myocardial infarction.
The relationship of marijuana use with coronary heart disease, including prognosis among patients with coronary heart disease, is uncertain. We conducted an inception cohort study of 1913 adults hospitalized with myocardial infarction at 45 US hospitals between 1989 and 1994, with a median follow-up of 3.8 years. We ascertained total mortality according to self-reported marijuana use in the preceding year. A total of 52 patients reported marijuana use during the prior year, and 317 patients died during follow-up. Compared with nonuse, marijuana use less than weekly was associated with a hazard ratio of 2.5 (95% CI, 0.9-7.3). The corresponding hazard ratio for weekly use or more was 4.2 (95% CI, 1.2-14.3). The age- and sex-adjusted hazard ratios associated with any use were 1.9 (95% CI, 0.6-6.3) for cardiovascular mortality and 4.9 (95% CI, 1.6-14.7) for noncardiovascular mortality. In a comparison of 42 marijuana users and 42 other patients matched on propensity scores, there were 6 deaths among marijuana users and one among non-users (log-rank P = .06). These preliminary results suggest possible hazards of marijuana for patients who survive acute myocardial infarction. Although marijuana use has not been associated with mortality in other populations, it may pose particular risk for susceptible individuals with coronary heart disease.


Predictive biomarkers in colorectal cancer: usage, validation, and design in clinical trials.
As cancer treatment development has shifted its attention to targeted therapies, it is becoming increasingly important to provide tools for selecting the right treatment for an individual patient to achieve optimal clinical benefit. Biomarkers, identified and studied in the process of understanding the nature of the disease at the molecular pathogenesis level, have been increasingly recognized as a critical aspect in more accurate diagnosis, prognosis assessment, and therapeutic targeting. Predictive biomarkers, which can aid treatment decisions, require extensive data for validation. In this article, we discuss the definition, clinical usages, and more extensively the clinical trial designs for the validation of predictive biomarkers. Predictive biomarker validation methods can be broadly grouped into retrospective and prospective designs. Retrospective validation utilizes data from previously conducted prospective randomized controlled trials. Prospective designs include enrichment designs, treatment-by-marker interaction designs, marker-based strategy designs, and adaptive designs. We discuss each design with examples and provide comparisons of the advantages and disadvantages among the different designs. We conclude that the combination of scientific, clinical, statistical, ethical, and practical considerations provides guidance for the choice of the clinical trial design for validation of each proposed predictive biomarker.


Brain adaptations of insulin signaling kinases, GLUT 3, p-BADser155 and nitrotyrosine expression in various hypoglycemic models of mice.
Insulin-induced moderate or severe hypoglycemia (MH or SH) impairs cognition and SH causes neuronal death. On the contrary, alternate day fasting (ADF) protects the brain during excitotoxic stress and improves cognitive function. Unlike the scenario in the periphery, insulin and its relationship towards brain glucose uptake and metabolism are considered to be less significant. Yet, the hypoglycemia associated brain metabolism is not clearly understood. The authors broadly investigated the brain metabolism in various hypoglycemic models such as insulin-induced MH, SH, SH with glucose reperfusion, 24 h fasting and ADF in the cortex or hippocampus of C57BL6/J mice. The authors analyzed the protein expression of insulin signaling kinases (plays a key role in neuronal survival and memory), Bcl-2 associated death promoter (p-BADser155) (dephosphorylation inhibits glucokinase activity and reduces glucose or increases ketone body metabolism in the brain), neuronal-specific glucose transporter 3 (GLUT 3) and nitrotyrosine (marker of nitric oxide which is involved in neuronal glucose uptake via GLUT 3) using western blotting analysis. Insulin-induced MH or SH differentially regulated the brain insulin signaling kinases. The expression of p-BADser155 decreased in all hypoglycemic models except the insulin-induced MH in hippocampus. The trended higher GLUT 3 and increased nitrotyrosine expression of insulin-induced SH were restored after glucose reperfusion. The trended higher or increased GLUT 3 and nitrotyrosine expression of ADF were positively correlated with serum beta-hydroxybutyrate levels. During hypoglycemia, it can be suggested that the brain might decrease glucose metabolism via glycolysis or prefer ketone body metabolism (except the insulin-induced MH in hippocampus) by modifying the p-BADser155 expression. In addition to the ketone body metabolism, the brain might adapt to uptake glucose in insulin-induced SH or ADF by modifying the GLUT 3 or nitrotyrosine expression.


Cross-priming in health and disease.
Cross-priming is an important mechanism to activate cytotoxic T lymphocytes (CTLs) for immune defence against viruses and tumours. Although it was discovered more than 25 years ago, we have only recently gained insight into the underlying cellular and molecular mechanisms, and we are just beginning to understand its physiological importance in health and disease. Here we summarize current concepts on the cross-talk between the immune cells involved in CTL cross-priming and on its role in antimicrobial and antitumour defence, as well as in immune-mediated diseases.


[A quantitative assessment of enzyme activity by cytophotometry using a standard].
The usage of the standard has been offered in cytospectrophotometrical measurements of enzymatic activities that are revealed histochemically. A tetrazolium method was used to demonstrate lactate dehydrogenase activity in sections of the rat brain. As a standard, a mixture of the enzyme (a crystalline forme of LDG) and gelatine was taken. The method of standardization permits fulfilling a quantitative comparison of heterogenic biological materials.