Cigarette smoke condensate alters Saccharomyces cerevisiae efflux transporter mRNA and activity and increases caffeine toxicity.
In animals, cigarette smoke may alter pharmacokinetics by altering activity and expression of ABC drug transporters. We previously demonstrated that cigarette smoke condensate (CSC) impairs activity and expression of several hepatic ABC drug transporters which mediate toxicant efflux. However, CSC effects on efflux transporters are still unknown in Saccharomyces cerevisiae which resists diverse chemical stresses, by inducing pleiotropic drug resistance (PDR) genes among others. The yeast ABC transporters are functionally and structurally homologous to the mammalian ones. In this study, Saccharomyces cerevisiae exposure to CSC for 15 min caused a dose-dependent inhibition of rhodamine 123 efflux, whereas a longer exposure (3 h) induced mRNA expression of the ABC PDR efflux pumps Pdr5, Snq2, Pdr 10 and Pdr15, and of Tpo1, a member of the major facilitator superfamily (MFS). CSC also increased toxicity of caffeine, which is handled by two PDR transporters, Pdr5 and Snq2. Taken together, these data demonstrated that yeast efflux transporters are targets of cigarette smoke chemicals, and that Saccharomyces cerevisiae may cope with CSC-induced stress, including the initial efflux inhibition, by induction of the mRNA of several plasma membrane PDR and MFS efflux transporters. Saccharomyces cerevisiae is therefore a valid model to investigate pollutant effects on ABC and MFS transporters.


Role of anchorin CII, a 31,000-mol-wt membrane protein, in the interaction of chondrocytes with type II collagen.
We have previously reported the isolation of a hydrophobic, type-II collagen-binding glycoprotein of molecular weight 31,000 (31,000-mol-wt protein) from chick chondrocyte membranes (Mollenhauer, J., and K. von der Mark, EMBO Eur. Mol. Biol. Organ. J., 2:45-50). The function of this protein in anchoring pericellular type II collagen to the chondrocyte surface was inferred from its ability to bind native type-II collagen either when detergent solubilized or when inserted into liposomes. In the present study we have used specific antibodies to localize this protein, which we now call anchorin CII, to the surface of chondrocytes in both cartilage sections, and in cell culture. In immunofluorescence studies of isolated chondrocytes we observed a dense, punctate distribution of anchorin CII on the cell surface when chondrocytes were enclosed by a pericellular type II collagen matrix. Removal of the pericellular matrix with trypsin also removed anchorin CII. The membrane protein character of anchorin CII was indicated by the demonstration of antibody-induced patching and capping on the chondrocyte surface at 22 degrees C and 37 degrees C, respectively. In monolayer culture, the amount of anchorin CII appeared reduced on flattened chondrocytes lacking a pericellular type II collagen matrix but was prominent upon intercellular cell processes. Fab' fragments prepared from either anchorin CII antiserum or an antiserum directed against the entire chondrocyte membrane inhibited the attachment of chondrocytes to a type II collagen substrate. In each case, the inhibition of attachment was neutralized by preincubation of Fab' fragments with purified anchorin CII.


Male gonadal function in coeliac disease: 1. Sexual dysfunction, infertility, and semen quality.
The prevalence of hypogonadism, sexual dysfunction and abnormalities of semen quality was determined in 28 consecutive males with coeliac disease. These observations were related to jejunal morphology and nutritional status, and were compared with findings in 19 men with Crohn's disease of similar age and nutritional status. Two of the 28 coeliacs (7%) had clinical evidence of hypogonadism but impotence and decreased sexual activity occurred more commonly, the latter apparently improving after gluten withdrawal. Of the married coeliacs, 19% had infertile marriages, a value greater than expected in the general population. Hypogonadism and sexual dysfunction were not detected in our patients with Crohn's disease. Seminal analysis in coeliacs revealed marked abnormalities of sperm morphology and motility, but only the former appeared to improve after gluten withdrawal. Similar abnormalities, however, were also detected in patients with Crohn's disease, although, unlike the coeliacs, 46% also had reduced concentrations of spermatozoa. Semen quality in coeliac disease could not be clearly related to general or specific (serum vitamin B(12) and red cell folate) nutritional deficiencies or to fertility, although sperm motility was markedly reduced in two of the three coeliacs with infertile marriages. The presence of antisperm antibodies did not appear to be an important aetiological factor in male infertility in coeliac disease. The pathogenesis of infertility and sexual dysfunction in coeliac disease remains unclear, suggesting that factors such as endocrine dysfunction or other specific nutritional deficiency may be involved.


Expression and characterization of an epoxide hydrolase from Anopheles gambiae with high activity on epoxy fatty acids.
In insects, epoxide hydrolases (EHs) play critical roles in the metabolism of xenobiotic epoxides from the food resources and in the regulation of endogenous chemical mediators, such as juvenile hormones. Using the baculovirus expression system, we expressed and characterized an epoxide hydrolase from Anopheles gambiae (AgEH) that is distinct in evolutionary history from insect juvenile hormone epoxide hydrolases (JHEHs). We partially purified the enzyme by ion exchange chromatography and isoelectric focusing. The experimentally determined molecular weight and pI were estimated to be 35 kD and 6.3 respectively, different than the theoretical ones. The AgEH had the greatest activity on long chain epoxy fatty acids such as 14,15-epoxyeicosatrienoic acids (14,15-EET) and 9,10-epoxy-12Z-octadecenoic acids (9,10-EpOME or leukotoxin) among the substrates evaluated. Juvenile hormone III, a terpenoid insect growth regulator, was the next best substrate tested. The AgEH showed kinetics comparable to the mammalian soluble epoxide hydrolases, and the activity could be inhibited by AUDA [12-(3-adamantan-1-yl-ureido) dodecanoic acid], a urea-based inhibitor designed to inhibit the mammalian soluble epoxide hydrolases. The rabbit serum generated against the soluble epoxide hydrolase of Mus musculus can both cross-react with natural and denatured forms of the AgEH, suggesting immunologically they are similar. The study suggests there are mammalian sEH homologs in insects, and epoxy fatty acids may be important chemical mediators in insects.


Transseptal puncture in left atrial appendage closure guided by 3D printing and multiplanar CT reconstruction.
The presented study investigates the application of bi-arterial 3D printed models to guide transseptal puncture (TSP) in left atrial appendage closure (LAAC). The objectives are to (1) test the feasibility of 3D printing (3DP) for TSP guidance, (2) analyse the distribution of the optimal TSP locations, and (3) define a CT-derived 2D parameter suitable for predicting the optimal TSP locations. Preprocedural planning included multiplanar CT reconstruction, 3D segmentation, and 3DP. TSP was preprocedurally simulated in vitro at six defined sites. Based on the position of the sheath, TSP sites were classified as optimal, suboptimal, or nonoptimal. The aim was to target the TSP in the recommended position during the procedure. Procedure progress was assessed post hoc by the operator. Of 68 screened patients, 60 patients in five centers (mean age of 74.68 ± 7.64 years, 71.66% males) were prospectively analyzed (3DP failed in one case, and seven patients did not finally undergo the procedure). In 55 patients (91.66%), TSP was performed in the optimal location as recommended by the 3DP. The optimal locations for TSP were postero-inferior in 45.3%, mid-inferior in 45.3%, and antero-inferior in 37.7%, with a mean number of optimal segments of 1.34 ± 0.51 per patient. When the optimal TSP location was achieved, the procedure was considered difficult in only two (3.6%) patients (but in both due to complicated LAA anatomy). Comparing anterior versus posterior TSP in 2D CCT, two parameters differed significantly: (1) the angle supplementary to the LAA ostium and the interatrial septum angle (160.83° ± 9.42° vs. 146.49° ± 8.67°; p = 0.001), and (2) the angle between the LAA ostium and the mitral annulus (95.02° ± 3.73° vs. 107.38° ± 6.76°; p < 0.001), both in the sagittal plane. In vitro TSP simulation accurately determined the optimal TSP locations for LAAC and facilitated the procedure. More than one-third of the optimal TSP sites were anterior.


Emotional intelligence and stress in medical students performing surgical tasks.
Poor stress management skills can compromise performance in the operating room, particularly in inexperienced trainees. Little is known about individual differences in managing stress. This study aimed to explore the relationship between trait emotional intelligence (EI) and objective and subjective measures of stress in medical students faced with unfamiliar surgical tasks. Seventeen medical undergraduates completed an unfamiliar laparoscopic task on a simulator during January to April 2008. Subjective stress before, during (retrospectively), and after the task was measured using the self-report State-Trait Anxiety Inventory. Objective stress was measured using continuous heart rate (HR) monitoring. Participants also completed the Trait Emotional Intelligence Questionnaire short form (TEIQue-SF). The authors computed scores for global trait EI and the TEIQue-SF four factors and carried out descriptive and correlational analyses. The highest levels of subjective stress were reported during the task and correlated positively with trait EI as well as with the trait EI factors of well-being and emotionality. Objective stress (mean HR) during the task was positively related to the sociability factor of trait EI. Higher trait EI scores were also associated with better after-task recovery from stress experienced during the task. Students with higher trait EI are more likely to experience stress during unfamiliar surgical scenarios but are also more likely to recover better compared with their lower-trait-EI peers. Trait EI has implications for the design of effective stress management training tailored to individual needs and potential applications to surgical trainee selection and development.


A study to investigate the evidential value of blue gel pen inks.
The aim of this project was to investigate the evidential value of blue gel pen inks in Europe. For this purpose, 33 blue gel pen inks, of different brands and models, representative of those available on the European market at the time of the study, were analyzed using three techniques: filtered light examination (FLE), Raman Spectroscopy (RS), and scanning electron microscopy (SEM). First, after visual examination (naked eye and stereo microscope), it was possible to classify the 33 inks into three groups described as: milky, metallic, and normal. This paper describes in detail the results obtained for the normal gel pen group. The ability of the techniques to discriminate gel inks between and within brands varied. The results indicated that RS and SEM were more discriminating than FLE. The greatest degree of differentiation was achieved when using a combination of RS and SEM techniques (discriminating power = 0.91). This study also highlights some problems concerning the identification of the brand of a gel pen from a written text.


"It wasn't working. " Women's experiences with short-term breastfeeding.
To investigate the lived experiences of women who initiate breastfeeding but stop within the first 2 weeks after birth. Phenomenology. Interviews were recorded on audio tape, transcribed verbatim, and analyzed for themes by an interdisciplinary group of researchers. Women described a clash or incongruity between highly idealized expectations and early breastfeeding problems. This led to incremental disillusionment and cessation of breastfeeding attempts. Women spoke poignantly about a sense of failure, guilt, or shame and had lingering self-doubts about not continuing with breastfeeding. Women need more accurate information about the process of initiating breastfeeding. Nurses should avoid teaching breastfeeding practices that can confuse infants and sabotage mothers and should work toward providing sensitive care that respects personal boundaries of mothers. Additionally, provisions need to be made for continued support and consultation after the mother's discharge from the hospital or birthing center. Finally, nurses and other health professionals should understand that women who stop breastfeeding at an early date may feel guilty about doing so, and may need assistance in resolving feelings about having made that choice.


The CSF accumulator: its role in the central nervous system and implications for advancing hydrocephalus shunt technology.
The 'cerebrospinal fluid (CSF) accumulator' plays an important physiological role in the healthy central nervous system as both a shock absorber and regulator of drainage. In the presence of hydrocephalus, this function is compromised by resistance to the flow of CSF. It is further compromised by the placement of a standard shunt which allows only for one-way directional diversion of the fluid. Complications associated with shunt dependency may be a direct consequence of both the condition, and a failure of existing systems to restore the normal accumulator function. This article reviews the pathophysiology of the 'CSF accumulator' as it relates to hydrocephalus and its management, and proposes a biomimetic shunt designed to restore the 'CSF accumulator' toward normal. It is hoped that such a device will support normalization of intracranial pressure and restore an appropriate level of CSF volume in the shunted patient.


Binocular disparity encoding cells generated through an Infomax based learning algorithm.
A learning algorithm for a model binocular cell was derived according to an information maximization principle and by using a low signal-to-noise-ratio approximation. The algorithm updates cell's synaptic weights so that the information obtained from the cell's output is increased. According to the algorithm, model binocular cells were trained by using computer-generated stereo images as training data. As a result, cells tuned to various disparities were generated. Also, generated synaptic weight patterns of the cells were similar to Gabor-wavelets and receptive fields of simple cells in the visual cortex. Thus, they were orientation and spatial frequency selective as well as disparity selective. Gabor functions were used to fit the generated weight patterns. The fitting results indicated that the generated cells encode disparities in terms of phase disparity and/or position disparity. This result agrees with experimental findings by Anzai et al. [J Neurophys 82 (1999) 874] and is consistent with ICA-based theoretical results obtained [Network: Comput Neural Syst 11 (2000) 191].


Rad50 ATPase activity is regulated by DNA ends and requires coordination of both active sites.
The Mre11-Rad50-Nbs1(Xrs2) (MRN/X) complex is critical for the repair and signaling of DNA double strand breaks. The catalytic core of MRN/X comprised of the Mre11 nuclease and Rad50 adenosine triphosphatase (ATPase) active sites dimerizes through association between the Rad50 ATPase catalytic domains and undergoes extensive conformational changes upon ATP binding. This ATP-bound 'closed' state promotes binding to DNA, tethering DNA ends and ATM activation, but prevents nucleolytic processing of DNA ends, while ATP hydrolysis is essential for Mre11 endonuclease activity at blocked DNA ends. Here we investigate the regulation of ATP hydrolysis as well as the interdependence of the two functional active sites. We find that double-stranded DNA stimulates ATP hydrolysis by hMRN over ∼20-fold in an end-dependent manner. Using catalytic site mutants to create Rad50 dimers with only one functional ATPase site, we find that both ATPase sites are required for the stimulation by DNA. MRN-mediated endonucleolytic cleavage of DNA at sites of protein adducts requires ATP hydrolysis at both sites, as does the stimulation of ATM kinase activity. These observations suggest that symmetrical engagement of the Rad50 catalytic head domains with ATP bound at both sites is important for MRN functions in eukaryotic cells.


The Association Between Interlimb Asymmetry and Athletic Performance Tasks: A Season-Long Study in Elite Academy Soccer Players.
Bishop, C, Read, P, Bromley, T, Brazier, J, Jarvis, P, Chavda, S, and Turner, A. The association between interlimb asymmetry and athletic performance tasks: A season-long study in elite academy soccer players. J Strength Cond Res 36(3): 787-795, 2022-The aims of this study were to determine the association between asymmetry and measures of speed and change of direction speed (CODS) performance throughout a competitive soccer season and, determine whether any observed changes in asymmetry were associated with changes in speed and CODS performance. Eighteen elite male under-23 academy soccer players performed unilateral countermovement jumps, unilateral drop jumps (DJ), 10- and 30-m sprints, and 505 CODS tests at pre, mid, and end of season. No significant relationships were evident during preseason or midseason between asymmetry and speed or CODS performance. Significant correlations were shown at the end of season between DJ height asymmetry and 10-m sprint time (ρ = 0.62; p = 0.006) and 505 time on the right limb (ρ = 0.65; p = 0.003). No significant correlations between changes in asymmetry and changes in speed or CODS were evident at any time point. Although numerous studies have reported associations between asymmetry and reduced athletic performance, it seems that these associations with speed and CODS do not track consistently over time. Thus, suggestions for the reduction of asymmetry that may indirectly enhance athletic performance cannot be made.


Telemedicine Coverage of Intensive Care Units: A Narrative Review.
Telemedicine coverage of intensive care units is an organizational innovation that has been touted as a means to improve access to and quality of critical care. The purpose of this narrative review is to discuss the different organizational models of intensive care unit telemedicine and factors that have influenced its adoption and to review the existing literature to consider whether it has lived up to its promise. We conclude by suggesting future directions to fill in some of the existing gaps in the literature.


Trajectories of alcohol use and problems during the COVID-19 pandemic: The role of social stressors and drinking motives for men and women.
Increased alcohol use coinciding with onset of the COVID-19 pandemic, particularly among women, has been documented among U.S. adults. This study examines trajectories of alcohol use and alcohol problems over a 9-month period during the pandemic, the extent to which these trajectories are predicted by social stress and drinking motives, and whether results differ for women and men. Data come from three online surveys of a nationally representative sample of U.S. adults ages 30-80 conducted in May-July 2020, October-November 2020, and March 2021. The analytic sample consists of N = 1118 who initially reported any past year alcohol use. The early-COVID survey assessed demographics, social stressors, and drinking motives. All three surveys assessed average drinks per day in past month and drinking-related problems. Alcohol use declined for men, but remained stable for women. Alcohol problems increased for both sexes, especially for men. Level of alcohol use was associated with loneliness and social demands for men, and drinking motives for both sexes, with changes in use related to loneliness and social demands for men. Level of alcohol problems was associated with loneliness for women and drinking motives for both sexes, with changes in problems related to drinking motives for women. Interactions of social stress with drinking motives were not found. Sex differences in alcohol use and alcohol problems during the pandemic-as well as their associations with indicators of social stress and drinking motives-highlight the importance of tailoring prevention and treatment efforts for men and women.


Quantitative analysis of dynamic protein-protein interactions in planta by a floated-leaf luciferase complementation imaging (FLuCI) assay using binary Gateway vectors.
Dynamic protein-protein interactions are essential in all cellular and developmental processes. Protein-fragment complementation assays allow such protein-protein interactions to be investigated in vivo. In contrast to other protein-fragment complementation assays, the split-luciferase (split-LUC) complementation approach facilitates dynamic and quantitative in vivo analysis of protein interactions, as the restoration of luciferase activity upon protein-protein interaction of investigated proteins is reversible. Here, we describe the development of a floated-leaf luciferase complementation imaging (FLuCI) assay that enables rapid and quantitative in vivo analyses of protein interactions in leaf discs floating on a luciferin infiltration solution after transient expression of split-LUC-labelled interacting proteins in Nicotiana benthamiana. We generated a set of eight Gateway-compatible split-LUC destination vectors, enabling fast, and almost fail-safe cloning of candidate proteins to the LUC termini in all possible constellations. We demonstrate their functionality by visualizing the well-established homodimerization of the 14-3-3 regulator proteins. Quantitative interaction analyses of the molybdenum co-factor biosynthesis proteins CNX6 and CNX7 show that the luciferase-based protein-fragment complementation assay allows direct real-time monitoring of absolute values of protein complex assembly. Furthermore, the split-LUC assay is established as valuable tool to investigate the dynamics of protein interactions by monitoring the disassembly of actin filaments in planta. The new Gateway-compatible split-LUC destination vector system, in combination with the FLuCI assay, provides a useful means to facilitate quantitative analyses of interactions between large numbers of proteins constituting interaction networks in plant cells.


Early detection of ecosystem regime shifts: a multiple method evaluation for management application.
Critical transitions between alternative stable states have been shown to occur across an array of complex systems. While our ability to identify abrupt regime shifts in natural ecosystems has improved, detection of potential early-warning signals previous to such shifts is still very limited. Using real monitoring data of a key ecosystem component, we here apply multiple early-warning indicators in order to assess their ability to forewarn a major ecosystem regime shift in the Central Baltic Sea. We show that some indicators and methods can result in clear early-warning signals, while other methods may have limited utility in ecosystem-based management as they show no or weak potential for early-warning. We therefore propose a multiple method approach for early detection of ecosystem regime shifts in monitoring data that may be useful in informing timely management actions in the face of ecosystem change.


Kernel-imbedded Gaussian processes for disease classification using microarray gene expression data.
Designing appropriate machine learning methods for identifying genes that have a significant discriminating power for disease outcomes has become more and more important for our understanding of diseases at genomic level. Although many machine learning methods have been developed and applied to the area of microarray gene expression data analysis, the majority of them are based on linear models, which however are not necessarily appropriate for the underlying connection between the target disease and its associated explanatory genes. Linear model based methods usually also bring in false positive significant features more easily. Furthermore, linear model based algorithms often involve calculating the inverse of a matrix that is possibly singular when the number of potentially important genes is relatively large. This leads to problems of numerical instability. To overcome these limitations, a few non-linear methods have recently been introduced to the area. Many of the existing non-linear methods have a couple of critical problems, the model selection problem and the model parameter tuning problem, that remain unsolved or even untouched. In general, a unified framework that allows model parameters of both linear and non-linear models to be easily tuned is always preferred in real-world applications. Kernel-induced learning methods form a class of approaches that show promising potentials to achieve this goal. A hierarchical statistical model named kernel-imbedded Gaussian process (KIGP) is developed under a unified Bayesian framework for binary disease classification problems using microarray gene expression data. In particular, based on a probit regression setting, an adaptive algorithm with a cascading structure is designed to find the appropriate kernel, to discover the potentially significant genes, and to make the optimal class prediction accordingly. A Gibbs sampler is built as the core of the algorithm to make Bayesian inferences. Simulation studies showed that, even without any knowledge of the underlying generative model, the KIGP performed very close to the theoretical Bayesian bound not only in the case with a linear Bayesian classifier but also in the case with a very non-linear Bayesian classifier. This sheds light on its broader usability to microarray data analysis problems, especially to those that linear methods work awkwardly. The KIGP was also applied to four published microarray datasets, and the results showed that the KIGP performed better than or at least as well as any of the referred state-of-the-art methods did in all of these cases. Mathematically built on the kernel-induced feature space concept under a Bayesian framework, the KIGP method presented in this paper provides a unified machine learning approach to explore both the linear and the possibly non-linear underlying relationship between the target features of a given binary disease classification problem and the related explanatory gene expression data. More importantly, it incorporates the model parameter tuning into the framework. The model selection problem is addressed in the form of selecting a proper kernel type. The KIGP method also gives Bayesian probabilistic predictions for disease classification. These properties and features are beneficial to most real-world applications. The algorithm is naturally robust in numerical computation. The simulation studies and the published data studies demonstrated that the proposed KIGP performs satisfactorily and consistently.


Glycyrrhizic Acid Inhibits High-Mobility Group Box-1 and Homocysteine-Induced Vascular Dysfunction.
Hyperhomocysteinemia (HHcy) worsens cardiovascular outcomes by impairing vascular function and promoting chronic inflammation via release of danger-associated molecular patterns, such as high-mobility group box-1 (HMGB-1). Elevated levels of HMGB-1 have recently been reported in patients with HHcy. Therefore, targeting HMGB-1 may be a potential therapy to improve HHcy-induced cardiovascular pathologies. This study aimed to further elucidate HMGB-1's role during acute HHcy and HHcy-induced atherogenesis and to determine if inhibiting HMGB-1 with glycyrrhizic acid (Glyz) improved vascular function. Male New Zealand White rabbits (n = 25) were placed on either a standard control chow (CD; n = 15) or atherogenic diet (AD; n = 10) for 4 weeks. Rabbit serum and Krebs taken from organ bath studies were collected to quantify HMGB-1 levels. Isometric tension analysis was performed on abdominal aorta (AA) rings from CD and AD rabbits. Rings were incubated with homocysteine (Hcy) [3 mM] for 60 min to induce acute HHcy or rhHMGB-1 [100 nM]. Vascular function was assessed by relaxation to cumulative doses of acetylcholine. Markers of vascular dysfunction and inflammation were quantified in the endothelium, media, and adventitia of AA rings. HMGB-1 was significantly upregulated in serum (p < 0.0001) and Krebs (p < 0.0001) after Hcy exposure or an AD. Incubation with Hcy (p < 0.0001) or rhHMGB-1 (p < 0.0001) and an AD (p < 0.0001) significantly reduced relaxation to acetylcholine, which was markedly improved by Glyz. HMGB-1 expression was elevated (p < 0.0001) after Hcy exposure and AD (p < 0.0001) and was normalized after Glyz treatment. Moreover, markers of vascular function, cell stress and inflammation were also reduced after Glyz. These results demonstrate that HMGB-1 has a central role during HHcy-induced vascular dysfunction and inhibiting it with Glyz could be a potential treatment option for cardiovascular diseases.


Low cost tuberculosis vaccine antigens in capsules: expression in chloroplasts, bio-encapsulation, stability and functional evaluation in vitro.
Tuberculosis (TB) caused by Mycobacterium tuberculosis is one of the leading fatal infectious diseases. The development of TB vaccines has been recognized as a major public health priority by the World Health Organization. In this study, three candidate antigens, ESAT-6 (6 kDa early secretory antigenic target) and Mtb72F (a fusion polyprotein from two TB antigens, Mtb32 and Mtb39) fused with cholera toxin B-subunit (CTB) and LipY (a cell wall protein) were expressed in tobacco and/or lettuce chloroplasts to facilitate bioencapsulation/oral delivery. Site-specific transgene integration into the chloroplast genome was confirmed by Southern blot analysis. In transplastomic leaves, CTB fusion proteins existed in soluble monomeric or multimeric forms of expected sizes and their expression levels varied depending upon the developmental stage and time of leaf harvest, with the highest-level of accumulation in mature leaves harvested at 6PM. The CTB-ESAT6 and CTB-Mtb72F expression levels reached up to 7.5% and 1.2% of total soluble protein respectively in mature tobacco leaves. Transplastomic CTB-ESAT6 lettuce plants accumulated up to 0.75% of total leaf protein. Western blot analysis of lyophilized lettuce leaves stored at room temperature for up to six months showed that the CTB-ESAT6 fusion protein was stable and preserved proper folding, disulfide bonds and assembly into pentamers for prolonged periods. Also, antigen concentration per gram of leaf tissue was increased 22 fold after lyophilization. Hemolysis assay with purified CTB-ESAT6 protein showed partial hemolysis of red blood cells and confirmed functionality of the ESAT-6 antigen. GM1-binding assay demonstrated that the CTB-ESAT6 fusion protein formed pentamers to bind with the GM1-ganglioside receptor. The expression of functional Mycobacterium tuberculosis antigens in transplastomic plants should facilitate development of a cost-effective and orally deliverable TB booster vaccine with potential for long-term storage at room temperature. To our knowledge, this is the first report of expression of TB vaccine antigens in chloroplasts.


[Thin layer chromatographic separation and enzymatic detection of carbamate pesticides in plant extracts].
The authors developed a thin-layer chromatographic technique for the separation of carbamate pesticides from plant material and an enzymatic method for their determination. The cellulose plates were impregnated with polar substances (ethylene glycol, triethylene glycol and beta,beta'-oxydipropionitrile). n-Heptane/chloroform and iso-octane/dioxane served as solvents. The slight enzyme-inhibiting effect and the colouration of the plant extracts render the detection of the pesticides difficult. Nevertheless, the separation and the enzymatic detection of Methomyl, Carbaryl, Baygon and Aldicarb could be achieved. The Rf values of the carbamates are not influenced by the kind (apples, pears, onions) and the quantity (50--200 mg) of the plant material.


Gene expression profiling of malignant mesothelioma cell lines: cDNA array study.
To reveal genes relevant for malignant mesothelioma (MM), we carried out cDNA array experiments on 4 MM cell lines and 2 primary mesothelial cell cultures established from pleural fluid of non-cancer patients. Human cancer gene filters including 588 genes were used for the cDNA array experiments. Our study revealed 26 over-expressed genes that play a role in the regulation of cell fate, cell cycle, cell growth and DNA damage repair and 13 under-expressed genes encoding growth factors, receptors and proteins involved in cell adhesion, motility and invasion to be common to 3 or 4 MM cell lines. We confirmed the cDNA array results using RT-PCR for 5 of the over-expressed genes and for 3 of the under-expressed genes. Our study presents gene expression profiles in MM cell lines and shows the involvement of several genes, such as those encoding JAGGED1, ser/thr protein kinase NIK, Ku80 and cyclin D2, novel in MM.


Metal uptake by woodlice in urban soils.
This study evaluated the uptake of bioavailable metal (Cd, Cr, Cu, Fe, Pb and Zn) by woodlice (Isopoda) collected from public open spaces in urban areas of Renfrewshire, Central West Scotland, UK. The species Oniscus asellus and Porcellio scaber were collected at 13 different locations together with associated surface soil samples. Soils were subject to sequential extraction to evaluate metal availability and analyzed by ICP-AES and flame AAS for Cd, Cr, Cu, Fe, Pb and Zn concentrations. The soil samples had metal concentrations typical of general urban environments and the potentially toxic elements were well below published guideline values for contaminated sites. The metal concentration showed differing inputs of natural and anthropogenic sources. Metals were bioconcentrated in the order Cu>Cd>Pb>Cr>Zn>Fe for O. asellus and Cu>Zn>Cd>Cr>Fe for P. scaber. Principal Component Analysis of soil geochemical properties and Isopoda metal concentration identified metal to metal variation in uptake. Multiple linear regression analysis was applied in order to investigate the metal uptake in relation to soil properties (total metal concentration, pH and organic matter (OM)). The results showed that factors affecting metal concentration were both species and site specific. The most available forms of metals were generally poorly related to metal accumulation by woodlice, with the only exception being for Cu, which was related to the exchangeable soil fraction. Soil conditions e.g. pH and OM, influenced metal association within the soil and OM played a significant role in restricting uptake of Cr and Pb in particular. For most of the metals studied, despite differences in the environmental availability of the metals, accumulation from ambient soil concentration is controlled by ecological and physiological factors influencing metal assimilation, storage and excretion and that the two biological species vary considerably in their regulation of individual metals.


Ginsenoside Rg1 Inhibits Glucagon-Induced Hepatic Gluconeogenesis through Akt-FoxO1 Interaction.
Rationale: Glucagon is involved in hepatic gluconeogenesis, playing a key role in type 2 diabetes. Ginsenosides are reported to have antidiabetic activities. Ginsenoside Rg1 is a major propanaxatriol-type saponin in ginseng. This study aims to investigate the regulatory effects of Rg1 on glucagon-induced hepatic glucose production. Methods: The effects of Rg1 were investigated in high-fat-diet (HFD)-fed mice and glucagon-challenged C57BL/6J mice. Glucose metabolism was evaluated by oral glucose tolerance test and pyruvate tolerance test. The role of Rg1 on the regulation of Akt-FoxO1 interaction was performed using immunofluorescence, immunoprecipitation, siRNA silencing, pharmacological inhibitor and active-site mutant in primary hepatocytes or HepG2 cells. Results: Abnormally elevated fasting glucagon levels were observed in HFD-fed mice, contributing significantly to increased fasting plasma glucose levels. Inappropriate fasting glucagon secretion inactivated Akt and promoted hepatic glucose production via upregulation of FoxO1 activity. Rg1 preserved glucagon-impaired Akt activation partly by binding to Akt at Ser473 site. Rg1 also promoted Akt binding to FoxO1 and inactivated FoxO1 by phosphorylation. Consequently, Rg1 decreased the hepatic glucose production through a decrease in transcription of phosphoenolpyruvate carboxykinase (PEPCK) and glucose 6-phosphatase (G6Pase). Both siRNA silencing of Akt and Akt inhibitor triciribine attenuated the effects of Rg1 in response to fasting hormone glucagon. Conclusion: Akt phosphorylation at Ser473 by ginsenoside Rg1 is critical for its gluconeogenesis-lowering effect, suggesting a potential for pharmaceutical intervention in response to fasting hormone glucagon.


Association between cannabis use and methadone maintenance treatment outcomes: an investigation into sex differences.
Cannabis will soon become legalized in Canada, and it is currently unclear how this will impact public health. Methadone maintenance treatment (MMT) is the most common pharmacological treatment for opioid use disorder (OUD), and despite its documented effectiveness, a large number of patients respond poorly and experience relapse to illicit opioids. Some studies implicate cannabis use as a risk factor for poor MMT response. Although it is well established that substance-use behaviors differ by sex, few of these studies have considered sex as a potential moderator. The current study aims to investigate sex differences in the association between cannabis use and illicit opioid use in a cohort of MMT patients. This multicentre study recruited participants on MMT for OUD from Canadian Addiction Treatment Centre sites in Ontario, Canada. Sex differences in the association between any cannabis use and illicit opioid use were investigated using multivariable logistic regression. A secondary analysis was conducted to investigate the association with heaviness of cannabis use. The study included 414 men and 363 women with OUD receiving MMT. Cannabis use was significantly associated with illicit opioid use in women only (OR = 1.82, 95% CI 1.18, 2.82, p = 0.007). Heaviness of cannabis use was not associated with illicit opioid use in men or women. This is the largest study to date examining the association between cannabis use and illicit opioid use. Cannabis use may be a sex-specific predictor of poor response to MMT, such that women are more likely to use illicit opioids if they also use cannabis during treatment. Women may show improved treatment outcomes if cannabis use is addressed during MMT.


Cycles of malaria associated with El Niño in Venezuela.
Malaria has been increasing globally, and epidemics tend to occur when weather conditions favor this vector-borne disease. Long-term meteorologic forecasting using El Niño Southern Oscillation (ENSO) may assist in anticipating epidemics and targeting scarce resources. To determine whether malaria epidemics in Venezuela are related to ENSO and rainfall and to determine whether such a relationship could be used to predict outbreaks. Retrospective analysis of national malaria morbidity (1975-1995) and mortality (1910-1935) data in the coastal zone and interior of Venezuela in relation to El Niño events and rainfall. Correlation between malaria mortality and morbidity and sea surface temperatures (SSTs) in the Eastern Tropical Pacific, a parameter of ENSO. Malaria mortality and morbidity have increased by an average of 36.5% (95% confidence interval, 3.7%-69.3%; P=.004) in years following recognized El Niño events. A moderate correlation was found between Pacific tropical SST during a Niño event and malaria 1 year later (r=0.50, P<.001). Malaria mortality is more strongly related to drought in the year preceding outbreaks than to rainfall during epidemic years. Historic and recent data from Venezuela demonstrate that malaria increases by an average of about one third in the year following a Nino event; change in malaria risk can be predicted from Pacific SSTs in the previous year. Therefore, the occurrence of an El Niño event may help predict malaria epidemics in this part of South America.


Analysis of global host gene expression during the primary phase of the Arabidopsis thaliana-Plasmodiophora brassicae interaction.
Microarray analysis was used to investigate changes in host gene expression during the primary stages of the interaction between the susceptible plant Arabidopsis thaliana (L.) Heynh ecotype Col-0 and the biotrophic pathogen Plasmodiophora brassicae Woronin. Analyses were conducted at 4, 7 and 10 days after inoculation (DAI) and revealed significant induction or suppression of a relatively low number of genes in a range of functional categories. At 4 DAI, there was induced expression of several genes known to be critical for pathogen recognition and signal transduction in other resistant host-pathogen interactions. As the pathogen further colonised root tissue and progressed through the primary plasmodium stage to production of zoosporangia at 7 and 10 DAI, respectively, fewer genes showed changes in expression. The microarray results were validated by examining a subset of induced genes at 4 DAI by quantitative real-time reverse transcriptase PCR (RT-qPCR) analysis all of which correlated positively with the microarray data. The two A. thaliana mutants jar1 and coiI tested were found to be susceptible to P. brassicae. The involvement of defence-related hormones in the interaction was further investigated and the findings indicate that addition of salicylic acid can suppress clubroot disease in A. thaliana plants.


Biomarkers of Alcohol Consumption in Body Fluids - Possibilities and Limitations of Application in Toxicological Analysis.
Ethyl alcohol is the most popular legal drug, but its excessive consumption causes social problems. Despite many public campaigns against alcohol use, car accidents, instances of aggressive behaviour, sexual assaults and deterioration in labor productivity caused by inebriated people is still commonplace. Fast and easy diagnosis of alcohol consumption is required in order to introduce proper and effective therapy, and is crucial in forensic toxicology analysis. The easiest method to prove alcohol intake is determination of ethanol in body fluids or in breath. However, since ethanol is rapidly metabolized in the human organism, only recent consumption can be detected using this method. Because of that, the determination of alcohol biomarkers was introduced for monitoring alcohol consumption over a wider range of time. The objective of this study was to review published studies focusing on the sample preparation methods and chromatographic or biochemical techniques for the determination of alcohol biomarkers in whole blood, plasma, serum and urine. An electronic literature search was performed to discuss possibilities and limitations of application of alcohol biomarkers in toxicological analysis. Authors described the markers of alcohol consumption such as: ethanol, its nonoxidative metabolites (ethyl glucuronide, ethyl sulfate, phosphatidylethanol, ethyl phosphate, fatty acid ethyl esters) and oxidative metabolites (acetaldehyde and acetaldehyde adducts). We also discussed issues concerning the detection window of these biomarkers, and possibilities and limitations of their use in routine analytical toxicology for monitoring alcohol consumption or sobriety during alcohol therapy.


Comparison Between Minimally Invasive Surgery and Conventional Open Surgery for Patients With Spinal Metastasis: A Prospective Propensity Score-Matched Study.
Prospective propensity score-matched study. To compare the outcomes of minimal invasive surgery (MIS) and conventional open surgery for spinal metastasis patients. There is lack of knowledge on whether MIS is comparable to conventional open surgery in treating spinal metastasis. Patients with spinal metastasis requiring surgery from January 2008 to December 2010 in two spine centers were recruited. The demographic, preoperative, operative, perioperative and postoperative data were collected and analyzed. Thirty MIS patients were matched with 30 open surgery patients using propensity score matching technique with a match tolerance of 0.02 based on the covariate age, tumor type, Tokuhashi score, and Tomita score. Both groups had significant improvements in Eastern Cooperative Oncology Group (ECOG), Karnofsky scores, visual analogue scale (VAS) for pain and neurological status postoperatively. However, the difference comparing the MIS and open surgery group was not statistically significant. MIS group had significantly longer instrumented segments (5.5 ± 3.1) compared with open group (3.8 ± 1.7). Open group had significantly longer decompressed segment (1.8 ± 0.8) than MIS group (1.0 ± 1.0). Open group had significantly more blood loss (2062.1 ± 1148.0 mL) compared with MIS group (1156.0 ± 572.3 mL). More patients in the open group (76.7%) needed blood transfusions (with higher average units of blood transfused) compared with MIS group (40.0%). Fluoroscopy time was significantly longer in MIS group (116.1 ± 63.3 s) compared with open group (69.9 ± 42.6 s). Open group required longer hospitalization (21.1 ± 10.8 days) compared with MIS group (11.0 ± 5.0 days). This study demonstrated that MIS resulted in comparable outcome to open surgery for patients with spinal metastasis but has the advantage of less blood loss, blood transfusions, and shorter hospital stay. 3.


The heart-gut axis: new target for atherosclerosis and congestive heart failure therapy.
The human gut microbiota has been identified as a possible novel risk factor for cardiovascular disease. The intestinal microbiome plays a role in the pathogenesis of atherosclerosis and heart failure. Even though studies in rodents suggested that gut microbes may affect the risk of heart disease, this link has not been shown in humans. In the present study, we review several potential mechanisms by which the gut microbiome and bacterial translocation are associated with the development of cardiac disorders making them potential targets for novel therapeutic measures for these conditions. Modulation of the gut microbiota as a mechanism for altering the pathogenesis of disorders is an area of growing interest. Alteration in the gut microbiota is being explored as a method of reducing risk factors associated with cardiac diseases.


A deficiency of lunatic fringe is associated with cystic dilation of the rete testis.
Lunatic fringe belongs to a family of beta1-3 N-acetyltransferases that modulate the affinity of the Notch receptors for their ligands through the elongation of O-fucose moieties on their extracellular domain. A role for Notch signaling in vertebrate fertility has been predicted by the intricate expression of the Notch receptors and their ligands in the oocyte and granulosa cells of the ovary and the spermatozoa and Sertoli cells of the testis. It has been demonstrated that disruption of Notch signaling by inactivation of lunatic fringe led to infertility associated with pleiotropic defects in follicle development and meiotic maturation of oocytes. Lunatic fringe null males were found to be subfertile. Here, we report that gene expression data demonstrate that fringe and Notch signaling genes are expressed in the developing testis and the intratesticular ductal tract, predicting roles for this pathway during embryonic gonadogenesis and spermatogenesis. Spermatogenesis was not impaired in the majority of the lunatic fringe null males; however, spermatozoa were unilaterally absent in the epididymis of many mice. Histological and immunohistochemical analysis of these testes revealed the development of unilateral cystic dilation of the rete testis. Tracer dye experiments confirm a block in the connection between the rete testis and the efferent ducts. Further, the dye studies demonstrated that many lunatic fringe mutant males had partial blocks of the connection between the rete testis and the efferent ducts bilaterally.


Cell-mediated immune response in primary biliary cirrhosis to a protein fraction from human bile.
Cell-mediated immune responses to a protein fraction of human bile have been demonstrated, using the leucocyte migration test, in eight out of 10 patients with primary biliary cirrhosis but in only three out of nine with active chronic hepatitis. In the latter condition sensitization to a liver-specific hepatocellular antigen was found more frequently (five out of nine patients) than in primary biliary cirrhosis (two out of 10). These results, as well as the granuloma formation observed histologically, suggest that the initial bile duct lesion in primary biliary cirrhosis may be associated with a cell-mediated response to antigens-perhaps derived from bile duct epithelial cells-which may be normal constituents of hepatic bile.


Clinical Application of Apheresis in Very Small Dogs Weighing <8 kg to Pediatric Patients.
Apheresis in low body weight children and adolescents is challenging due to a variety of technical and clinical issues including vascular access, low total blood volume, and hypotension. Although dogs have been a valuable preclinical model for apheresis, the procedure can be challenging since many pure-bred dogs are extremely small. Therefore, apheresis in these very small breeds presents very similar challenges as seen when performing the procedure in very low body weight people. We describe apheresis of four very small dogs, weighing from 4.6 to 7.6 kg, using either a COBESpectra and Spectra Optia apheresis system (Terumo BCT, Lakewood, CO, USA). Two dogs underwent large volume leukapheresis to collect mononuclear cells in preparation for hematopoietic stem cell transplantation and two dogs underwent therapeutic plasma exchange to treat an immune-mediated disease. In all cases, a dual-lumen hemodialysis catheter placed in the jugular vein provided adequate machine inlet and return flow rates. Machine priming was necessary to maintain hemodynamic stability during the beginning of the procedure, and rinseback was avoided for the same reason. Anticoagulant citrate dextrose solution, solution A was used for the large volume leukapheresis procedures and a combination of anticoagulant citrate dextrose solution, solution A and heparin was used for the therapeutic plasma exchange procedures. As such, serum iCa levels were regularly monitored and 10% calcium gluconate constant rate infusions were used to prevent citrate toxicity. All dogs completed the aphereses with no life-threatening adverse events. We conclude that aphereses in very small dogs is feasible if close attention is paid to hemodynamic stability and citrate toxicity.


Do therapists address gender and power in infidelity? A feminist analysis of the treatment literature.
Sociocontextual factors such as gender and power play an important role in the etiology of affairs and in recovery from them, yet it is unclear how current treatment models address these issues. Drawing on feminist epistemology, this study utilized a grounded theory analysis of 29 scholarly articles and books on infidelity treatment published between 2000 and 2010 to identify the circumstances under which gender and power issues were or were not part of treatment. We found five conditions that limit attention to gender and power: (a) speaking (or assuming) as though partners are equal, (b) reframing infidelity as a relationship problem, (c) limiting discussion of societal context to background, (d) not considering how societal gender and power patterns impact relationship dynamics, and (e) limiting discussion of ethics on how to position around infidelity. Analysis explored how each occurred across three phases of couple therapy. The findings provide a useful foundation for a sociocontextual framework for infidelity treatment.


Different Expression of Extracellular Signal-Regulated Kinases (ERK) 1/2 and Phospho-Erk Proteins in MBA-MB-231 and MCF-7 Cells after Chemotherapy with Doxorubicin or Docetaxel.
Curative treatment of breast cancer patients using chemotherapy often fails as a result of intrinsic or acquired resistance of the tumor to the drug. ERK is one of the main components of the Ras/Raf/MEK/ERK cascade, which mediates signal from cell surface receptors to transcription factors to regulate different gene expression. In this study, cytotoxicity and the expression of Erk1/2 and phospho-ERK was compared in MDA-MB-231 (ER-) and MCF-7 (ER+) cell lines after treatment with doxorubicin (DOX) or docetaxel (DOCT). Cell cytotoxicity of DOX or DOCT was calculated using MTT assay. Immonofluorescent technique was used to show MDR-1 protein in MDA-MB-231 and MCF-7 cells after treatment with DOX or DOCT. The expression of ERK1/2 and phpspho-ERK was assayed with immunoblotting. Comparing IC50 values showed that MDA-MB-231 cells are more sensitive than MCF-7 cells to DOX or DOCT. Immonofluorescent results confirmed the expression of MDR-1 in these two cell lines after DOX or DOCT treatment. In MDA-MB-231 cells the expression of ERK1/2 and phospho-ERK was decreased after DOX treatment in a dose-dependent manner. In contrast in MCF-7 cells the expression of ERK1/2 and phospho-ERK was increased after DOX treatment. DOCT treatment demonstrated the same result with less significant differences than DOX. The heterogeneity seen in cell lines actually reflects the heterogeneity of breast cancers. That is why, patients categorized in one group respond differently to a single treatment. These results emphasize the importance of a more accurate classification and a more specific treatment of breast cancer subtypes.


Automated live cell imaging of green fluorescent protein degradation in individual fibroblasts.
To accurately interpret the data from fluorescent proteins as reporters of gene activation within living cells, it is important to understand the kinetics of the degradation of the reporter proteins. We examined the degradation kinetics over a large number (>1,000) of single, living cells from a clonal population of NIH3T3 fibroblasts that were stably transfected with a destabilized, enhanced green fluorescent protein (eGFP) reporter driven by the tenascin-C promoter. Data collection and quantification of the fluorescence protein within a statistically significant number of individual cells over long times (14 h) by automated microscopy was facilitated by culturing cells on micropatterned arrays that confined their migration and allowed them to be segmented using phase contrast images. To measure GFP degradation rates unambiguously, protein synthesis was inhibited with cycloheximide. Results from automated live cell microscopy and image analysis indicated a wide range of cell-to-cell variability in the GFP fluorescence within individual cells. Degradation for this reporter was analyzed as a first order rate process with a degradation half-life of 2.8 h. We found that GFP degradation rates were independent of the initial intensity of GFP fluorescence within cells. This result indicates that higher GFP abundance in some cells is likely due to higher rates of gene expression, because it is not due to systematically lower rates of protein degradation. The approach described in this study will assist the quantification and understanding of gene activity within live cells using fluorescent protein reporters.


First Episode Psychosis Following COVID-19 Vaccination: a Case Report.
Vaccines are crucial to ending the COVID-19 pandemic. An mRNA-based COVID-19 vaccine can cause mild to moderate side effects. A number of cases of cardiac, gastrointestinal, and psychiatric side effects have been reported as rare side effects associated with the COVID-19 vaccine. This article presents a patient, who after the second injection of the mRNA-based COVID-19 vaccine, immediately developed anxiety, nonspecific fear, and insomnia as the prodromal phase of psychosis. Starting from the second week, the patient manifested delusions of persecution, delusions of influence, thoughts insertion, and delusional behaviour, culminating in the suicide attempt. The duration of psychosis was eight weeks, and symptom reduction was observed only after the gradual administration of antipsychotics over four weeks. The investigations of the patient did not support any structural changes of the brain, any severe medical conditions, a neurological abnormality, a confusion or a state of unconsciousness or alterations in laboratory tests. Psychosis due to the use of alcohol or psychoactive substances was excluded. The psychological assessment of the patient demonstrated the endogenous type of thinking, and the patient had schizoid and paranoid personality traits strongly associated with schizophrenia. This case indicates a strong causal relationship between the mRNA-based COVID-19 vaccine injection and the onset of psychosis. We intend to follow up this case for possible development of schizophrenia and understand that the COVID-19 vaccine could possible play a trigger role in the development of primary psychosis. Longer-term supporting evidence is needed to estimate the prevalence of psychosis following vaccination with the mRNA-based COVID-19 vaccine.


Liver involvement in patients with Gaucher disease types I and III.
Gaucher disease (GD) is a multisystemic disease. Liver involvement in GD is not well characterised and ranges from hepatomegaly to cirrhosis and hepatocellular carcinoma. We aim to describe, and assess the effect of treatment, on the hepatic phenotype of a cohort of patients with GD types I and II. Retrospective study based on the review of the medical files of the Gaucher Reference Centre of the Hospital de Clínicas de Porto Alegre, Brazil. Data from all GD types I and III patients seen at the centre since 2003 were analysed. Variables were compared as pre- ("baseline") and post-treatment ("follow-up"). Forty-two patients (types I: 39, III: 3; female: 22; median age: 35 y; enzyme replacement therapy: 37; substrate reduction therapy: 2; non-treated: 3; median time on treatment-MTT: 124 months) were included. Liver enzyme abnormalities, hepatomegaly, and steatosis at baseline were seen in 19/28 (68%), 28/42 (67%), and 3/38 patients (8%), respectively; at follow-up, 21/38 (55%), 15/38 (39%) and 15/38 (39%). MRI iron quantification showed overload in 7/8 patients (treated: 7; MTT: 55 months), being severe in 2/7 (treated: 2/2; MTT: 44.5 months). Eight patients had liver biopsy (treated: 6; MTT: 58 months), with fibrosis in 3 (treated: 1; time on treatment: 108 months) and steatohepatitis in 2 (treated: 2; time on treatment: 69 and 185 months). One patient developed hepatocellular carcinoma. GD is a heterogeneous disease that causes different patterns of liver damage even during treatment. Although treatment improves the hepatocellular damage, it is associated with an increased rate of steatosis. This study highlights the importance of a follow-up of liver integrity in these patients.


Design, manufacture and initial tests of a drug-eluting coronary stent.
A drug-eluting coronary stent is being developed at the National Institute of Cardiology of Mexico for the treatment of ischemic heart disease. To establish the best animal model for the tests, to show the advances in the drug-eluting stent prototype, to assess two drugs' antiproliferative activity and histological results. Smooth muscle cell culture tests were performed in order to assess sirolimus and paclitaxel antiproliferative properties. The drugs were encapsulated inside the polymeric matrix of the stents. Rabbits and pigs were used as animal models. Sirolimus and paclitaxel showed an inhibitory effect, which was higher for the latter. Infrared spectroscopy and light and optical microscopy showed that the drug/polymer layer properly adhered to the stent. At a four-week follow-up, both animal models showed satisfactory clinical evolution and adequate histological response, although the porcine model was shown to be more suitable for future protocols. Preliminary tests of the drug-eluting stent provided bases for the development of a study protocol with an adequate number of pigs and with clinical angiographic and histopathological three-month follow-up.


[Primary Leishmania infantum MON-80 endonasal leishmaniasis in Tunisia].
Mucocutaneous leishmaniasis is endemic in Central and South America. It causes massive mutilating and disfiguring lesions and can lead to destruction of facial structures. In Tunisia, leishmaniasis of the mucous membranes is rare, usually developing as a complication of cutaneous leishmaniasis via direct extension. We report the first case in Tunisia of isolated and primary nasal leishmaniasis. A 70-year-old man with a history of nephrectomy for renal lithiasis was seen with a painless nodule that had been present for one month. The latter was erythematous, polypoid and firm, with a diameter of 2 cm, and was situated in the right endonasal mucosa. The diagnosis of leishmaniasis was confirmed by histological and direct examinations revealing high numbers of amastigotes of Leishmania. Culture of the offending organism in NNN medium and isoenzymatic characterization resulted in identification of MON-80 Leishmania infantum leishmaniasis. The outcome was good with treatment, and the nodule was deflated after six months. There have been few reports of similar cases of primary and isolated mucosal leishmaniasis caused by Leishmania infantum. Our case is also unusual in that zymodeme MON-80 is only rarely a cause of Mediterranean leishmaniasis.


Diffusing capacity of lungs for carbon monoxide associated with subclinical myocardial impairment in systemic sclerosis: A cardiac MR study.
Systemic sclerosis (SSc) is characterised by microvascular and fibrotic lesions, which are located not only in skin but also in lungs and heart. This study aimed to investigate the association between lung function and myocardial T1 values using cardiac MR (CMR) imaging in patients with SSc without cardiovascular symptoms. The SSc patients and age- and sex-matched healthy subjects underwent CMR. The cardiac function and native T1 values of myocardium and lung function were measured. Spearman's rank correlations and linear regression analyses were performed to determine the association between lung function and myocardial T1. Forty-five SSc patients (aged 47.7±13.2 years, 40 females) and 23 (aged 46.0±14.4 years, 20 females) healthy subjects were enrolled. SSc patients exhibited considerably higher native T1 values compared with healthy subjects (1305.9±49.8 ms vs 1272.6±37.6 ms, p=0.006). Linear regression analysis revealed that decrease of diffusing capacity of lungs for carbon monoxide (DLCO) in SSc patients was notably associated with myocardial native T1 value before (β -1.017; 95% CI -1.883 to -0.151; p=0.022) and after adjusting for confounding factors (β -1.108; 95% CI -2.053 to -0.164; p=0.023). Moderate-to-severe decrease of DLCO was found to be significantly associated with myocardial native T1 value (β 48.006; 95% CI 17.822 to 78.190; p=0.003) after adjusting for confounding factors. DLCO inversely correlates with myocardial native T1 values in SSc patients, particularly moderate-to-severely decreased DLCO, suggesting that DLCO might be a potential indicator for subclinical myocardial impairment in SSc patients.


Parameters to enhance human hybridoma formation with hypoosmolar electrofusion.
Hypoosmolar conditions have permitted the development of electrofusion techniques capable of producing human hybridomas from as few as 10(5) B cells. A hybridoma formation efficiency of one hybrid for each 125 input B cells has been achieved with Epstein-Barr virus--activated B cells and mouse-human heteromyelomas. This is at least 100-fold higher in efficiency than with polyethylene glycol-induced cell fusion, as well as a 50- to 100-fold decrease in the required number of human B cells. The ability to fuse a small number of input B cells should lead to a greater success rate in immortalizing the rare antigen-specific B cells. The critical parameters include fusion voltages, the composition and number of wash steps used in cell preparation, the composition and duration of exposure to hypoosmolar fusion medium, fusion ratio, plating density, the use of growth medium without pH indicator, and the use of an irradiated human fibroblast feeder layer. By manipulating these parameters, a high hybrid yield can be achieved with different mouse-human heteromyelomas and Epstein-Barr virus-activated B cells.


Low taurine, gamma-aminobutyric acid and carnosine levels in plasma of diabetic pregnant rats: consequences for the offspring.
Gestational diabetes compromises fetal development and induces a diabetogenic effect in the offspring, including the development of gestational diabetes and the transmission of the effect to the next generation. Changes are not limited to glucose and insulin metabolism, and appear to be modulated by alterations at the hypothalamo-hypophyseal axis. In the present work, serum concentrations are given for the non-protein amino-acids taurine and gamma-aminobutyric acid (GABA), both neurotransmitters essential for normal brain development, and for the endogenous neuroprotector carnosine, a known anti-oxydans. Taurine levels are significantly below normal values in mildly diabetic mothers, in their fetal and adult offspring, virgin and pregnant, and in the fetuses of these pregnant offspring. GABA and carnosine levels are at the limit of detection in the diabetic mothers and their offspring at every stage. It is concluded that the low taurine, GABA and carnosine levels in diabetic mothers and their fetuses might compromise the normal structural and functional development of the fetal brain. When adult, these offspring present a deficiency of the circulating levels of these neurotransmitters involved in the hypothalamo-hypophyseal regulation of insulin secretion. This might contribute to the development of impaired glucose tolerance and gestational diabetes, thereby transmitting the effect to the next generation.


Oleuropein, chief constituent of olive leaf extract, prevents the development of morphine antinociceptive tolerance through inhibition of morphine-induced L-type calcium channel overexpression.
It has been shown that blockade of L-type calcium channels could abolish the development of opioid-induced antinociceptive tolerance. Here, the antitolerant effects of olive leaf extract (OLE) and its main component, oleuropein, which have a calcium channel blocker property were determined. Adult male Wistar rats were injected with morphine (20 mg/kg, i.p.) for 8 days to induce antinociceptive tolerance. Then OLE (50-200 mg/kg i.g.) and oleuropein (1-10 mg/kg i.p.) were injected concomitantly with morphine. The tail-flick test was used to assess the nociceptive threshold. The dorsal half of the lumbar spinal cord was assayed for the expression of L-type calcium channel using semiquantitative RT-PCR. The results showed that OLE (200 mg/kg) completely prevented morphine tolerance development. In addition, oleuropein in dose of 10 mg/kg, but not in 5 mg/kg, prevented the development of morphine antinociceptive tolerance. In addition, a significant increase in the mRNA levels of calcium channel (43.9%) was observed in the lumbar spinal cord of tolerant animals, which was reversed by effective of dose OLE. In conclusion, the results indicate that olive leaf extract has a potential antitolerant property against the chronic usage of morphine and that its main component, oleuropein, is responsible for such effect.


Changes in alcohol involvement, cognitions and drinking and driving behavior for youth after they obtain a driver's license.
This study tested whether obtaining a driver's license was associated with increases in alcohol and other drug involvement and changes in alcohol-related cognitions for youth, and whether drinking and driving behavior increased with driving experience. Confidential, anonymous surveys were conducted at two time points (fall, spring) with students at four high schools in San Diego county (N = 2,865, 51% female). Data were collected on alcohol, cigarette and marijuana use, license status, alcohol use by peers, attitudes towards drinking and driving and drinking and driving behaviors. Nondrivers (60%), new drivers (obtained a license between Time 1 and Time 2) and experienced drivers (26%) were compared on study variables at both time points and over time. Initially obtaining a driver's license was associated with increased frequency of substance use. Results were not significant for quantity of alcohol use, frequency of heavy drinking or perceived alcohol use norms. Attitudes towards drinking and driving reflected an increase in the perceived dangerousness of this behavior for new drivers. Drinking and driving behavior during the last 30 days increased with increased driving experience. The results indicate a number of changes in substance involvement after obtaining a driver's license. However, initially this transition may also indicate a period of protection against drinking and driving. These results may have implications for the target and content of drinking and driving interventions.


Research on teaching methods.
Research on teaching methods in nursing education was categorized into studies on media, CAI, and other nontraditional instructional strategies. While the research differed, some generalizations may be made from the findings. Multimedia, whether it is used for individual or group instruction, is at least as effective as traditional instruction (lecture and lecture-discussion) in promoting cognitive learning, retention of knowledge, and performance. Further study is needed to identify variables that may influence learning and retention. While learner attitudes toward mediated instruction tended to be positive, investigators failed to control for the effect of novelty. Control over intervening variables was lacking in the majority of studies as well. Research indicated that CAI is as effective as other teaching methods in terms of knowledge gain and retention. Attitudes toward CAI tended to be favorable, with similar problems in measurement as those evidenced in studies of media. Chang (1986) also recommends that future research examine the impact of computer-video interactive instruction on students, faculty, and settings. Research is needed on experimental teaching methods, strategies for teaching problem solving and clinical judgment, and ways of improving the traditional lecture and discussion. Limited research in these areas makes generalizations impossible. There is a particular need for research on how to teach students the diagnostic reasoning process and encourage critical thinking, both in terms of appropriate teaching methods and the way in which those strategies should be used. It is interesting that few researchers studied lecture and lecture-discussion except as comparable teaching methods for research on other strategies. Additional research questions may be generated on lecture and discussion in relation to promoting concept learning, an understanding of nursing and other theories, transfer of knowledge, and development of cognitive skills. Few studies attempted to identify variables that may influence learning, particularly characteristics of the learner. Only six investigators addressed learning styles and their interactions with the teaching method and outcomes (Gillies, 1984; Goldsmith, 1984; Kirchhoff & Holzemer, 1979; Kissinger & Munjas, 1981; Norris, 1986; Stein et al., 1972). Research in the future needs to focus on the relationship of different learner characteristics, attributes of the teaching method, and learning outcomes. In addition, initial learning, retention, transfer to practice, and instructional time should be studied. Characteristics of the teacher and setting and relationship to the methodologies used and outcomes of instruction need investigation. Research should attempt to identify optimal conditions for learning and ways in which methods should be used for particular students, subject matter, and points in the nursing curriculum...


A Comprehensive Quality Evaluation Method Based on C30-HPLC and an Analytic Hierarchy Process for the Chinese Herbal Formula, Erzhiwan.
The quantitative analysis of multiple indexes remains an important quality evaluation method of traditional Chinese medicine (TCM) herbal formulas. The Chinese Pharmacopoeia 2015 only stipulates the content of a single component, specnuezhenide, in Erzhiwan composed of the Fructus Ligustri Lucidi (FLL) powder and aqueous extracts of Herba Ecliptae (HE). To generalize the intrinsic quality of Erzhiwan, a novel C30-HPLC method with good precision, accuracy, and reproducibility was developed for the simultaneous determination of six compounds, including two isomers, and then an analytic hierarchy process was further applied to integrate and discriminate the quality of four samples prepared via different methods. The results of the analysis were in agreement with the antioxidant tests in vitro. This comprehensive strategy could provide a reference and suggestions for the improvement of the quality evaluation method of TCM herbal formulas.


IGF2BP family of RNA-binding proteins regulate innate and adaptive immune responses in cancer cells and tumor microenvironment.
Insulin-like growth factor 2 mRNA-binding proteins (IGF2BP1, IGF2BP2, and IGF2BP3) are a family of RNA-binding proteins that play an essential role in the development and disease by regulating mRNA stability and translation of critical regulators of cell division and metabolism. Genetic and chemical inhibition of these proteins slows down cancer cell proliferation, decreases invasiveness, and prolongs life span in a variety of animal models. The role of RNA-binding proteins in the induction of tissues' immunogenicity is increasingly recognized, but, the impact of the IGF2BPs family of proteins on the induction of innate and adaptive immune responses in cancer is not fully understood. Here we report that downregulation of IGF2BP1, 2, and 3 expression facilitates the expression of interferon beta-stimulated genes. IGF2BP1 has a greater effect on interferon beta and gamma signaling compared to IGF2BP2 and IGF2BP3 paralogs. We demonstrate that knockdown or knockout of IGF2BP1, 2, and 3 significantly potentiates inhibition of cell growth induced by IFNβ and IFNγ. Mouse melanoma cells with Igf2bp knockouts demonstrate increased expression of MHC I (H-2) and induce intracellular Ifn-γ expression in syngeneic T-lymphocytes in vitro. Increased immunogenicity, associated with Igf2bp1 inhibition, "inflames" mouse melanoma tumors microenvironment in SM1/C57BL/6 and SW1/C3H mouse models measured by a two-fold increase of NK cells and tumor-associated myeloid cells. Finally, we demonstrate that the efficiency of anti-PD1 immunotherapy in the mouse melanoma model is significantly more efficient in tumors that lack Igf2bp1 expression. Our retrospective data analysis of immunotherapies in human melanoma patients indicates that high levels of IGF2BP1 and IGF2BP3 are associated with resistance to immunotherapies and poor prognosis. In summary, our study provides evidence of the role of IGF2BP proteins in regulating tumor immunogenicity and establishes those RBPs as immunotherapeutic targets in cancer.


NbCZF1, a Novel C2H2-Type Zinc Finger Protein, as a New Regulator of SsCut-Induced Plant Immunity in Nicotiana benthamiana.
SsCut, which functions as an elicitor, can induce plant immunity. In this study, we utilized Nicotiana benthamiana and virus-induced gene silencing to decrease the expression of > 2,500 genes individually. Using this forward genetics approach, several genes were identified that, when silenced, compromised SsCut-triggered cell death based on a cell death assay. A C2H2-type zinc finger gene was isolated from N. benthamiana Sequence analysis indicated that the gene encodes a 27 kDa protein with 253 amino acids containing two typical C2H2-type zinc finger domains; this gene was named NbCZF1 We found that SsCut-induced cell death could be inhibited by virus-induced gene silencing of NbCZF1 in N. benthamiana In addition, SsCut induces stomatal closure, accompanied by reactive oxygen species (ROS) production by NADPH oxidases and nitric oxide (NO) production. NbCZF1-silenced plants showed impaired SsCut-induced stomatal closure, decreased SsCut-induced production of ROS and NO in guard cells and reduced SsCut-induced resistance against Phytophthora nicotianae Taken together, these results demonstrate that the NbCZF1-ROS-NO pathway mediates multiple SsCut-triggered responses, including stomatal closure, hypersensitive responses and defense-related gene expression. This is the first report describing the function of a C2H2-type zinc finger protein in N. benthamiana.


Food ingredient safety evaluation: Utility and relevance of toxicokinetic methods.
The use of toxicokinetic (TK) data is becoming more prevalent in the evaluation of food ingredient safety as more TK information is being incorporated in safety data packages. Data demonstrating "1) the extent of absorption, 2) tissue distribution, 3) pathways and rates of metabolism, and 4) rate(s) of elimination" of food ingredients and their metabolites of intermediate and high toxicological potential may be useful for planning and designing toxicity studies, selecting doses for toxicity studies, addressing species differences, and understanding the potential modes of action to evaluate their safety. TK data reported in the literature or generated from mechanistic TK studies can be analyzed using mathematical methods, including compartment and noncompartment TK methods, whose predictions can enhance interpretation of observed effects. Because of recent advancements, several approaches have been developed to improve sensitivity of analyses of available TK data and reduce uncertainty for evaluating safety of food ingredients. An example of advanced TK methods is physiologically-based TK (PBTK) modeling that incorporates physiological/biochemical parameters into a TK framework to predict internal exposure. In this review, we discuss the utility of some TK methods and explore their relevance and potential value for food ingredient safety evaluation. We also describe the strengths and limitations of these TK methods and discuss current challenges and opportunities for expanding their application for evaluating safety of food ingredients. This review represents a state of science report, and not a guidance document, on the utility and relevance of TK methods for the safety evaluation of food ingredients.


Deep Time Paleoproteomics: Looking Forward.
The goal of paleoproteomics is to characterize proteins from specimens that have been subjected to the degrading and obscuring effects of time, thus obtaining biological information about tissues or organisms both unobservable in the present and unobtainable through morphological study. Although the description of sequences from Tyrannosaurus rex and Brachylophosaurus canadensis suggested that proteins may persist over tens of millions of years, the majority of paleoproteomic analyses have focused on historical, archeological, or relatively young paleontological samples that rarely exceed 1 million years in age. However, recent advances in methodology and analyses of diverse tissues types (e.g., fossil eggshell, dental enamel) have begun closing the large window of time that remains unexplored in the fossil history of the Cenozoic. In this perspective, we discuss the history and current state of deep time paleoproteomics (DTPp), here defined as paleoproteomic study of samples ∼1 million years (1 Ma) or more in age. We then discuss the future of DTPp research, including what we see as critical ways the field can expand, advancements in technology that can be utilized, and the types of questions DTPp can address if such a future is realized.


Pharmacologic management of trigeminal nerve injury pain after dental implant surgery.
Injuries to the trigeminal nerve are a common postoperative complication of dental implant surgery. Usually, the altered sensation and neuropathic pain caused by the nerve injury is temporary, but a permanent neurosensory disorder can sometimes occur. Surgery is commonly used to treat this condition, but the treatment is associated with some complications and a relatively low success rate. This study analyzed the characteristics of pharmacologic management of trigeminal nerve injury pain after dental implant surgery. Eighty-five patients who visited a temporomandibular joint and orofacial pain clinic with a history of trigeminal nerve injury pain after dental implant surgery were enrolled in this study. The pharmacologic management for trigeminal nerve injury pain was evaluated by prescribing a variety of medications for 12 weeks according to the prescription protocol of the study. The patients' pain characteristics, average percentage of pain reduction, and pain relieving factors were investigated prospectively. Patients who took anticonvulsants and antidepressants for at least 12 weeks reported a mean reduction in pain of 24.8%. Interestingly, patients who experienced an altered sensation and neuropathic pain for more than 1 year also reported a reduction in pain and discomfort, with an average decrease of 17.1%. In addition, it was found that early treatment using medications had a significant effect on reducing the level of pain and discomfort. These results suggest that pharmacologic management can be used for treating trigeminal nerve injury pain after dental implant surgery.


Magnetic resonance imaging in the diagnosis and treatment of a canine spinal cord injury.
A nine-year-old female crossbred dog was presented with tetraplegia following a fall. No vertebral abnormalities were detectable on plain radiographs of the cervical spine. Magnetic resonance imaging scans revealed absence of extraparenchymal compression and an area of oedema within the cervical spinal cord, suggesting a favourable prognosis. Following nursing care and physiotherapy, the dog recovered the ability to walk, although mild neurological deficits persisted in the left limbs.


Recommendations for the use of cardiac troponin measurement in acute cardiac care.
The release of cardiomyocyte components, i.e. biomarkers, into the bloodstream in higher than usual quantities indicates an ongoing pathological process. Thus, detection of elevated concentrations of cardiac biomarkers in blood is a sign of cardiac injury which could be due to supply-demand imbalance, toxic effects, or haemodynamic stress. It is up to the clinician to determine the most probable aetiology, the proper therapeutic measures, and the subsequent risk implied by the process. For this reason, the measurement of biomarkers always must be applied in relation to the clinical context and never in isolation. There are a large number of cardiac biomarkers, but they can be subdivided into four broad categories, those related to necrosis, inflammation, haemodynamic stress, and/or thrombosis. Their usefulness is dependent on the accuracy and reproducibility of the measurements, the discriminatory limits separating pathology from physiology, and their sensitivity and specificity for specific organ damage and/or disease processes. In recent years, cardiac biomarkers have become important adjuncts to the delivery of acute cardiac care. Therefore, the Working Group on Acute Cardiac Care of the European Society of Cardiology established a committee to deal with ongoing and newly developing issues related to cardiac biomarkers. The intention of the group is to outline the principles for the application of various biomarkers by clinicians in the setting of acute cardiac care in a series of expert consensus documents. The first of these will focus on cardiac troponin, a pivotal marker of cardiac injury/necrosis.


TEM investigation of formation mechanism of monocrystal-thick b-oriented pure silica zeolite MFI film.
The first direct transmission electron microscopic (TEM) observation has been carried out on the continuous monocrystal-thick b-oriented pure silica zeolite MFI films produced by in situ crystallization. The self-supporting film samples for TEM study were fabricated by dissolving the steel substrate with acid. This TEM study is free of those artifacts that are typically associated with TEM sample preparations, and allows us to investigate the "true" structure and texture of a very large area of the film and at the same time to focus at will on each individual zeolite crystal in the film. Abundant TEM information including crystallographic orientation relationships among crystals in the film (both out-of-plane and in-plane), grain boundaries, and each crystal grain was obtained. This TEM investigation provides direct unambiguous new evidence to support the homogeneous nucleation mechanism, by which the films form through homogeneous nucleation and crystal growth in the bulk to form equal-sized disk-shape crystals, followed by self-assembly of these crystals onto the substrate to produce a two-dimensional close-packed structure. The last stage of the film formation involves simultaneous space-limited growth and rotation of the individual crystals to realize the in-plane crystallographic control within the film.


Improvement of endocytoscopic findings after per oral endoscopic myotomy (POEM) in esophageal achalasia; does POEM reduce the risk of developing esophageal carcinoma? Per oral endoscopic myotomy, endocytoscopy and carcinogenesis.
Per oral endoscopic myotomy (POEM) has been reported to be a new therapeutic option for esophageal achalasia. The possibility that POEM could reduce the risk of developing esophageal squamous cell carcinoma was evaluated. This was a single-centre, retrospective study. Fifteen consecutive patients with esophageal achalasia who underwent POEM in our institution between August 2010 and January 2012 were enrolled. Ultra-high magnification with endocytoscopy was performed, and both histopathological and immunohistochemical evaluations for Ki-67 and p53 were assessed before and 3 months after POEM. POEM was successfully performed and effectively released the dysphagia symptom in all patients without severe complications. Subjective symptoms (mean Ekcardt score, before 7.4 vs. after 0.5, p<0.05) and manometric pressure studies (mean lower esophageal sphincter pressure), before 82.7 vs. after 22.9 mmHg, p<0.05) showed substantial improvement following POEM. The average numbers of esophageal epithelial nuclei before and after POEM on endocytoscopic images were 128.0 and 78.0, respectively (p<0.05). The mean Ki-67-positive ratio was 26.0 (median 25.4, range, 10.3-33.2) before and 20.7 (median 20.0, 13.1-29.9; p=0.07) after POEM, and the mean p53-positive ratio was 2.35 (median 2.61, 0.32-4.23) before and 0.97 (median 1.49, 0.32-1.56; p<0.05) after POEM. A significant positive correlation was seen between the number of nuclei and the Ki-67-positive ratio (p<0.05). POEM appears to be an effective and less invasive treatment of choice against achalasia and may reduce the risk of esophageal carcinogenesis. Endocytoscopy can be useful for the assessment of esophageal cellular proliferation.


Enrichment of CFU-S proliferation inhibitor-producing cells based on their identification by the monoclonal antibody F4/80.
Cells capable of synthesizing a factor that can specifically inhibit the proliferation of haemopoietic spleen colony-forming units, CFU-S, carry many of the characteristics of macrophages. The monoclonal antibody, F4/80, which is specific for murine macrophages, has therefore been used to isolate macrophages from bone marrow. These macrophages were then assayed for their ability to produce the inhibitor. Low-density bone marrow cells were first separated by a density-cut procedure and then labelled with F4/80. Fluorescence-activated cell sorting was then used to select F4/80 positive and negative cell fractions. It was found that the F4/80 positive fraction contains inhibitor cells, enriched by at least 200-fold compared to unfractionated marrow. Subsequent culture of this fraction over a period of several weeks produced a further 15- to 20-fold increase in inhibitor-producing capacity. The cultured cells producing inhibitor were virtually 100% F4/80 positive, phagocytic and exhibited histochemical properties characteristic of macrophages. The macrophage-like character of the producer cells was thus confirmed and the removal of the majority of unwanted cells means that many of the impurities in the normal crude extracts have been excluded.


Direct-acting antiviral treatment for chronic hepatitis C in people who use drugs in a real-world setting.
Direct-acting antivirals (DAAs) offer high cure rates in people who inject drugs (PWID) with hepatitis C virus (HCV) infection. There are concerns regarding lower response rates among PWID in real life. We evaluated the outcome of DAA therapy in PWID in a real-world setting and the factors that affect it. We performed a retrospective analysis of 174 PWID with chronic hepatitis C who started DAAs in a Greek liver clinic in collaboration with an addiction program. Patients who did not return for reassessment were considered as lost to follow up (LTFU). A logistic regression model was used to assess factors associated with a sustained virological response 12 weeks after treatment completion (SVR12) and LTFU. Patients' mean age was 48±9.2 years and 91/174 (52.3%) were attending opioid substitution treatment programs. Overall, 144/174 (82.8%) patients completed therapy and presented for SVR12 testing, 8/174 (4.6%) did not complete treatment and 22/174 (12.6%) were LTFU. Overall SVR12 was 79.9% (139/174). For those with an available SVR12 test the response rate reached 96.5% (139/144). Regression analysis did not indicate any significant association between patient characteristics and SVR12. Age <45 years and genotype 3 were independent predictors of LTFU. Parallel use was found to have a trend towards LTFU. HCV treatment by hepatologists and addiction specialists is feasible, effective and safe in a real-world setting. However, as 12% of patients appear to be LTFU, more emphasis should be placed on interventions guaranteeing follow up for SVR testing and general care.


Translational research on parenting of adolescents: Linking theory to valid observation measures for family centered prevention and treatment.
Parental monitoring and family problem solving are key parenting practices targeted in evidence-based interventions targeting adolescents and families, yet the constructs have yet to be validated across ethnic groups. The study's objective was to promote translational research by evaluating convergent, discriminant, and predictive validity of the two constructs at age 16-17 years through the use of multiple observation indicators and methods and as a function of ethnic status. Videotaped parent-adolescent family interactions were coded for monitoring and problem solving in a sample of 714 European American (EA; 59.2 %) and African American (AA; 40.8 %) males (53.8 %) and females (46.2 %). Structural equation models established convergent and discriminant validity of parental monitoring and problem solving among parent, youth, and observation measures for AA and EA families. Low levels of parent monitoring was highly predictive of antisocial behavior in EA and in AA youths (p < 0.001) and moderately predicted future drug use (p < 0.001) for both groups at age 18-19. Poorer family problem solving was also moderately predictive of antisocial behavior (p < 0.001 for EA; p < 0.05 for AA) and drug use (p < 0.01 for EA; p < 0.05 for AA) at age 18-19. These findings suggest that interventions targeting parental monitoring and family problem solving can be reliably evaluated through various measurement methods and that such interventions are of value in efforts to prevent and treat problem behavior in adolescence. These family processes are readily observable in videotaped family interaction tasks in both EA and AA families.


Studies of the occupational exposure of Malaysian plantation workers to paraquat.
Studies carried out on the occupational exposure to paraquat of plantation workers in Malaysia comprised quantitative estimates of dermal and respiratory exposure of knapsack spray operators, carriers, and rubber tappers operating under their normal working conditions. Spray operators have been shown to be dermally exposed to paraquat by walking through recently sprayed vegetation and into their own spray, regular adjustment and unblocking of spray nozzles and leakage, and overfilling of knapsack spray tanks. Carriers also received measurable dermal exposure from walking through recently sprayed vegetation and accidental spillage when carrying and loading. The infrequent and negligible dermal exposure of tappers resulted from walking through recently sprayed vegetation. Determinations of the total airborne paraquat concentrations in the breathing zone show that spray operators and carriers are exposed to an order of 1% or less of the current TLV for respirable paraquat. No paraquat was detected in the breathing zones of tappers working in simultaneously sprayed blocks. The calculated ranges of dermal and respiratory exposures, when compared with published data on both the exposure to, and the toxicity of, paraquat, indicate that there should be no toxicological risk to any of the three groups studied as a result of using paraquat.


Treatment of cardiac arrest in the hyperbaric environment: key steps on the sequence of care--case reports.
The U.S. territory of Guam attracts thousands of military and civilian divers annually and is home to the only recompression facility within a 5,000-km radius that accepts critically injured dive casualties. As recompression chambers are confined spaces and standard use of electrical cardioversion cannot be used, cardiac resuscitation at depth must deviate from advanced cardiovascular life support (ACLS) algorithms. Furthermore, many hyperbaric chambers that accept dive casualties are in remote locations, a situation that requires providers to approach cardiac resuscitation in a different way when compared to an in-hospital or ICU setting. This presents a challenge to trained medical and diving professionals. We present two contrasting vignettes of diving injuries initially responsive to appropriate treatment but then deteriorating during recompression therapy and ultimately requiring resuscitation at depth. Additionally, we explore the physiologic basis of resuscitation in a hyperbaric environment as it relates to the treatment of cardiac arrest at depth. This review critically examines the current guidelines in place for emergency cardiac resuscitation in a hyperbaric chamber followed by recommendations for the key steps in the sequence of care.


Dabigatran etexilate: a novel oral thrombin inhibitor for thromboembolic disease.
To evaluate the efficacy and safety of dabigatran etexilate, approved by the Food and Drug Administration (FDA) in October 2010 for the prevention of cardioembolic stroke in patients with atrial fibrillation; potential off-label use is treatment and prevention of venous thromboembolic disorders. Literature was accessed through MEDLINE (1977-April 2011) and International Pharmaceutical Abstracts (1977-April 2011) using the terms dabigatran, dabigatran etexilate, BIBR 1048, direct thrombin inhibitor, anticoagulant, and thromboembolism. In addition, US government Web sites, including clinicaltrials.gov and fda.gov, were reviewed for pertinent information. Lastly, reference citations from publications identified in the initial search were reviewed. All English-language articles identified from the data sources were evaluated. For the evaluation of clinical efficacy and safety, only Phase 2 and 3 studies are included in this review. In 6 published Phase 3 trials to date, dabigatran has exhibited a similar efficacy and safety profile to that of comparator drugs, including dose-adjusted warfarin and enoxaparin, at various dosages. In the largest of these trials, RE-LY (Randomized Evaluation of Long-Term Anticoagulation Therapy), dabigatran was at least as effective as dose-adjusted warfarin in reducing stroke or systemic embolism. Overall bleeding risks were similar; however, dabigatran may be associated with a lower incidence of intracranial bleeding and hemorrhagic stroke but a higher incidence of gastrointestinal bleeding. Although dabigatran is not approved for venous thromboembolism (VTE) prevention or treatment, results of the RE-MODEL and RE-NOVATE trials suggest similar efficacy to once-daily dosing of enoxaparin 40 mg but inferior efficacy to the FDA-approved twice-daily dosing of enoxaparin 30 mg in the RE-MOBILIZE trial. Dabigatran is an effective and safe alternative to oral vitamin K antagonists for stroke prevention in patients with nonvalvular atrial fibrillation, with fewer drug interactions and monitoring requirements. Additionally, dabigatran may be a viable alternative to enoxaparin in VTE prevention and warfarin in VTE treatment, although current trial data are limited.