Differential tissue distribution of pharmaceuticals in a wild subtropical marine fish.

To date, the presence of pharmaceuticals has been extensively documented across a wide range of aquatic systems and biota. Further, substantial progress has been made in transitioning from laboratory assessments of pharmaceutical fate and effects in fish to in situ assessments of exposure and effects; however, certain research areas remain understudied. Among these is investigation of differential accumulation across multiple internal tissues in wild marine fish beyond the species commonly sampled in laboratory and freshwater field settings. This study examined the presence of pharmaceuticals across four tissues (plasma, muscle, brain, and liver) in a wild marine fish, bonefish (Albula vulpes), throughout coastal South Florida, USA. Differential accumulation across tissues was assessed for the number and concentration, identity, and composition of accumulated pharmaceuticals by sampling 25 bonefish and analyzing them for 91 pharmaceuticals. The concentration of pharmaceuticals was highest in plasma > liver > brain > muscle, while the number of pharmaceuticals was highest in liver > brain > plasma > muscle. The identity of detected pharmaceuticals was tissue specific, and there was an inverse relationship between the number of detections for each pharmaceutical and its log Kow. The composition of pharmaceuticals was tissue specific for both pharmaceutical presence/absence and concentration. Across all tissues, the greatest similarity was between brain and liver, which were more similar to plasma than to muscle, and muscle was the most distinct tissue. For tissue compositional variability, muscle was the most diverse in accumulated pharmaceuticals, while plasma, brain, and liver were similarly variable. With the highest concentrations in plasma and highest number in liver, and documented variability in accumulated pharmaceuticals across tissues, our results highlight the importance of tissue selection when surveying exposure in wild fish, suggesting that multi-tissue analysis would allow for a more comprehensive assessment of exposure diversity and risk of adverse effects.

Identifying pathways of pharmaceutical exposure in a mesoconsumer marine fish.

Pharmaceutical uptake involves processes that vary across aquatic systems and biota. However, single studies examining multiple environmental compartments, microhabitats, biota, and exposure pathways in mesoconsumer fish are sparse. We investigated the pharmaceutical burden in bonefish (Albula vulpes), pathways of exposure, and estimated exposure to a human daily dose. To evaluate exposure pathways, the number and composition of pharmaceuticals across compartments and the bioconcentration in prey and bonefish were assessed. To evaluate bioaccumulation, we proposed the use of a field-derived bioaccumulation factor (fBAF), due to variability inherent to natural systems. Exposure to a human daily dose was based on bonefish daily energetic requirements and consumption rates using pharmaceutical concentrations in prey. Pharmaceutical number and concentration were highest in prey, followed by bonefish, water and sediment. Fifteen pharmaceuticals were detected in common among bonefish, prey, and water; all of which bioconcentrated in prey and bonefish, and four bioaccumulated in bonefish. The composition of detected pharmaceuticals was compartment specific, and prey were most similar to bonefish. Bonefish were exposed to a maximum of 1.2 % of a human daily dose via prey consumption. Results highlight the need for multicompartment assessments of exposure and consideration of prey along with water as a pathway of exposure.

Treatment With a Patented 3.6:1 Myo-Inositol to D-chiro-Inositol Ratio, Antioxidants, Vitamins and Minerals Food Supplement in Women With a History of Assisted Reproductive Technique (ART) Failures: A Series of Case Reports.

Infertility affects 15% of couples in reproductive age worldwide. In women in particular, infertility can be caused by various abnormalities, with polycystic ovary syndrome (PCOS) being the most common. Currently, there are many assisted reproductive techniques (ART) available to combat the burden of infertility. However, positive results are not guaranteed. The administration of inositol has been shown to increase positive reproductive outcomes in women undergoing ART. Here we present a series of clinical cases in which women with a history of infertility and previously failed ART, supplemented with a specific 3.6:1 MYO:DCI ratio, antioxidants, vitamins, and minerals for a period of 1 to 3 months before undergoing in vitro fertilization (IVF). In this series of case reports, we provide preliminary evidence that supplementation with a specific 3.6:1 MYO to DCI ratio, as well as antioxidants, vitamins, and minerals may contribute positively to female fertility in women undergoing IVF, with a history of primary or secondary infertility and previously failed ART.

Understanding pharmaceutical exposure and the potential for effects in marine biota: A survey of bonefish (Albula vulpes) across the Caribbean Basin.

Most research on pharmaceutical presence in the environment to date has focused on smaller scale assessments of freshwater and riverine systems, relying mainly on assays of water samples, while studies in marine ecosystems and of exposed biota are sparse. This study investigated the pharmaceutical burden in bonefish (Albula vulpes), an important recreational and artisanal fishery, to quantify pharmaceutical exposure throughout the Caribbean Basin. We sampled 74 bonefish from five regions, and analyzed them for 102 pharmaceuticals. We assessed the influence of sampling region on the number of pharmaceuticals, pharmaceutical assemblage, and risk of pharmacological effects. To evaluate the risk of pharmacological effects at the scale of the individual, we proposed a metric based on the human therapeutic plasma concentration (HTPC), comparing measured concentrations to a threshold of 1/3 the HTPC for each pharmaceutical. Every bonefish had at least one pharmaceutical, with an average of 4.9 and a maximum of 16 pharmaceuticals in one individual. At least one pharmaceutical was detected in exceedance of the 1/3 HTPC threshold in 39% of bonefish, with an average of 0.6 and a maximum of 11 pharmaceuticals exceeding in a Key West individual. The number of pharmaceuticals (49 detected in total) differed across regions, but the risk of pharmacological effects did not (23 pharmaceuticals exceeded the 1/3 HTPC threshold). The most common pharmaceuticals were venlafaxine (43 bonefish), atenolol (36), naloxone (27), codeine (27), and trimethoprim (24). Findings suggest that pharmaceutical detections and concentration may be independent, emphasizing the need to monitor risk to biota regardless of exposure diversity, and to focus on risk quantified at the individual level. This study supports the widespread presence of pharmaceuticals in marine systems and shows the utility of applying the HTPC to assess the potential for pharmacological effects, and thus quantify impact of exposure at large spatial scales.

Muscle secreted factors enhance activation of the PI3K/Akt and ß-catenin pathways in murine osteocytes.

Skeletal muscle and bone interact at the level of mechanical loading through the application of force by muscles to the skeleton and more recently focus has been placed on molecular/biochemical coupling of these two tissues. We sought to determine if muscle and muscle-derived factors were essential to the osteocyte response to loading. Botox® induced muscle paralysis was used to investigate the role of muscle contraction during in vivo tibia compression loading. 5-6 month-old female TOPGAL mice had their right hindlimb muscles surrounding the tibia injected with either BOTOX® or saline. At four days post injections when muscle paralysis peaked, the right tibia was subjected to a single session of in vivo compression loading at ∼2600 µÎµ. At 24 h post-load we observed a 2.5-fold increase in ß-catenin signaling in osteocytes in the tibias of the saline injected mice, whereas loading of tibias from Botox® injected mice failed to active ß-catenin signaling in osteocytes. This suggests that active muscle contraction produces a factor(s) that is necessary for or conditions the osteocyte's ability to respond to load. To further investigate the role of muscle derived factors, MLO-Y4 osteocyte-like cells and a luciferase based ß-catenin reporter (TOPflash-MLO-Y4) cell line we developed were treated with conditioned media (CM) from C2C12 myoblasts (MB) and myotubes (MT) and ex vivo contracted Extensor Digitorum Longus (EDL) and Soleus (Sol) muscles under static or loading conditions using fluid flow shear stress (FFSS). 10 % C2C12 myotube CM, but not myoblast or NIH3T3 fibroblast cells CM, induced a rapid activation of the Akt signaling pathway, peaking at 15 min and returning to baseline by 1-2 h under static conditions. FFSS applied to MLO-Y4 cells for 2 h in the presence of 10 % MT-CM resulted in a 6-8 fold increase in pAkt compared to a 3-4 fold increase under control or when exposed to 10 % MB-CM. A similar response was observed in the presence of 10 % EDL-CM, but not in the presence of 10 % Sol-CM. TOPflash-MLO-Y4 cells were treated with 10 ng/ml Wnt3a in the presence or absence of MT-CM. While MT-CM resulted in a 2-fold activation and Wnt3a produced a 10-fold activation, the combination of MT-CM + Wnt3a resulted in a 25-fold activation of ß-catenin signaling, implying a synergistic effect of factors in MT-CM with Wnt3a. These data provide clear evidence that specific muscles and myotubes produce factors that alter important signaling pathways involved in the response of osteocytes to mechanical load. These data strongly suggest that beyond mechanical loading there is a molecular coupling of muscle and bone.

Early transcranial direct current stimulation with modified constraint-induced movement therapy for motor and functional upper limb recovery in hospitalized patients with stroke: A randomized, multicentre, double-blind, clinical trial.

BACKGROUND: Constraint-induced movement therapy (CIMT) and transcranial direct current stimulation (tDCS) are used to reduce interhemispheric imbalance after stroke, which is why the combination of these therapies has been used for neurological recovery, but not in the acute phase. OBJECTIVES: To evaluate the effectiveness of combining active or sham bihemispheric tDCS with modified CIMT (mCIMT) for the recovery of the Upper Limb (UL) in hospitalized patients with acute and subacute stroke. METHODS: This randomized controlled, double-blind, placebo-controlled, parallel group clinical trial was executed between September 2018 to March 2021 recruited 70 patients. The patients were randomized to one of two groups to receive treatment for 7 consecutive days, which included 20 min of active or sham bihemispheric tDCS daily (anodal ipsilesional and cathodal contralesional), with an mCIMT protocol. The primary outcome was the difference in the evolution of motor and functional upper limb recovery with assessment on days 0, 5, 7, 10 and 90. The secondary outcomes were independence in activities of daily living (ADL) and quality of life. RESULTS: The active group presented a statistically significant gap compared to the simulated group throughout the trend in the scores of the FMA (motor function and joint pain) and WMFT (functional ability and weight to box) (p < 0.05) and showed a minimal clinically important difference (FMA: difference between groups of 4.9 points [CI: 0.007- 9.799]; WMFT: difference between groups of 6.54 points [CI: 1.10-14.15]). In the secondary outcomes, there was a significant difference between the groups in ADL independence (Functional Independence Measure: difference of 8.63 [CI: 1.37-18.64]) and perceived recovery of quality of life evaluated at 90 days (p = 0.0176). CONCLUSIONS: Combining mCIMT with bihemispheric tDCS in patients hospitalized with acute-subacute stroke allows us to maximize the motor and functional recovery of the paretic upper limb in the early stages and independence in ADL, maintaining the effects over time.

Water temperature affects the biotransformation and accumulation of a psychoactive pharmaceutical and its metabolite in aquatic organisms.

Pharmaceutically active compounds (PhACs) have been shown to accumulate in aquatic and riparian food-webs. Yet, our understanding of how temperature, a key environmental factor in nature, affects uptake, biotransformation, and the subsequent accumulation of PhACs in aquatic organisms is limited. In this study, we tested to what extent bioconcentration of an anxiolytic drugs (temazepam and oxazepam) is affected by two temperature regimes (10 and 20 °C) and how the temperature affects the temazepam biotransformation and subsequent accumulation of its metabolite (oxazepam) in aquatic organisms. We used European perch (Perca fluviatilis) and dragonfly larvae (Sympetrum sp.), which represent predator and prey species of high ecological relevance in food chains of boreal and temperate aquatic ecosystems. Experimental organisms were exposed to target pharmaceuticals at a range of concentrations (0.2-6 µg L-1) to study concentration dependent differences in bioconcentration and biotransformation. We found that the bioconcentration of temazepam in perch was significantly reduced at higher temperatures. Also, temperature had a strong effect on temazepam biotransformation in the fish, with the production and subsequent accumulation of its metabolite (oxazepam) being two-fold higher at 20 °C compared to 10 °C. In contrast, we found no temperature dependency for temazepam bioconcentration in dragonfly larvae and no detectable biotransformation of the parent compound that would result in measurable concentrations of oxazepam in this organism. Our results highlight that while organisms may share the same aquatic ecosystem, their exposure to PhACs may change differently across temperature gradients in the environment.

Multi-link endoscopic manipulator robot actuated by shape memory alloys spring actuators controlled by a sliding mode.

The aim of this study was to design and evaluate a prototype of a snake-like endoscopic manipulator robot (SLEMR) and its corresponding automatic controller based on the first order sliding mode theory. The SLEMR was controlled with a set of actuators made of shape memory alloys (SMA). The SLEMR device was constructed with a sequential arrangement of links interconnected by a two degree-of-freedom joint. A parallel agonist-antagonist configuration of actuators was implemented to move each joint. The physical relation between temperature and elongation in SMA forced the execution of the movement in the joint. Elongation-temperature model of the SMA actuator served to get a feasible bound of velocity for each joint. Each pair of SMA actuators was controlled by a first order sliding mode controller. This control design solved the tracking trajectory problem for each joint in the SLEMR because of its robustness against uncertainties and external perturbations. The control action was projected into a feasible implementable set of pulse-width modulated signals which was used to regulate the temperature of the corresponding SMA actuator. The controller designed in this study was experimentally validated in a SLEMR made up by a tridimensional printing technique. The control strategy induced the successful trajectory tracking for all the joints in the SLEMR simultaneously. This characteristic of the control design also enforces the tracking of a reference position by the tip of the final link of the SLEMR. An image acquisition system was used to determine the position of the final actuator in the SLEMR. The effectiveness of the controller proposed in this study was confirmed by the evaluation of the tracking error of the final actuator which approached to a bounded region (less than 1.0 mm) near the origin in a finite-time (0.5 s).

Following the infection dynamics of the tropical trematode Oligogonotylus mayae in its intermediate and definitive hosts for 13 years.

We present a time series of 13 years (2003-2016) of continuous monthly data on the prevalence and mean abundance of the trematode Oligogonotylus mayae for all the hosts involved in its life cycle. We aimed to determine whether annual (or longer than annual) environmental fluctuations affect these infection parameters of O. mayae in its intermediate snail host Pyrgophorus coronatus, and its second and definitive fish host Mayaheros urophthalmus from the Celestun tropical coastal lagoon, Yucatan, Mexico. Fourier time series analysis was used to identify infection peaks over time, and cross-correlation among environmental forcings and infection parameters. Our results suggest that the transmission of O. mayae in all its hosts was influenced by the annual patterns of temperature, salinity and rainfall. However, there was a biannual accumulation of metacercarial stages of O. mayae in M. urophthalmus, apparently associated with the temporal range of the El Niño-Southern Oscillation (five years) and the recovery of the trematode population after a devasting hurricane. Taking O. mayae as an example of what could be happening to other trematodes, it is becoming clear that environmental forcings acting at long-term temporal scales affect the population dynamics of these parasites.

Physical inactivity and excessive sucrose consumption are associated with higher serum lipids in subjects with Taq1B CETP polymorphism.

BACKGROUND: Dyslipidaemias result from the interaction between genetic and environmental factors, including diet disequilibrium and physical inactivity. Among the genetic factors associated with serum lipids, the Taq1B CETP polymorphism has been investigated. The B1 allele has been considered as a risk factor for dyslipidaemia because of its association with greater CETP levels and higher serum triglycerides. The present study aimed to determine the role of the Taq1B polymorphism with lipid and anthropometric variables and its interaction with diet and physical activity. METHODS: In total, 215 subjects were enrolled in this cross-sectional study. Diet intake was evaluated using a 3-day food consumption record and physical activity was determined in accordance with World Health Organization recommendations. The Taq1B CETP polymorphism was determined by allelic discrimination. RESULTS: Subjects with the B1B2/B2B2 genotype, who had a sucrose consumption ≥5% of the total kcal day-1 , had higher levels of total cholesterol (TC) [165.55 (142.21-188.89) mg dL-1 versus 200.19 (184.79-215.60) mg dL-1 ; P for interaction = 0.034] and low-density lipoprotein [99.29 (75.52-123.05) mg dL-1 versus 128.64 (113.59-143.69) mg dL-1 ; P for interaction = 0.037] than subjects with the B1B1 genotype. Subjects who did not perform physical activity and had the B1B2/B2B2 genotype showed significantly higher levels of TC [177.48 (161.36-193.60) mg dL-1 versus 194.49 (185.43-203.56) mg mL-1 ; P for interaction = 0.033] than subjects with the B1B1 genotype. CONCLUSIONS: We provide evidence that subjects with inadequate environmental factors carriers of the polymorphic genotype had higher serum lipid levels than subjects with the B1B1 genotype.

Impact of cooking and nixtamalization on the bioaccessibility and antioxidant capacity of phenolic compounds from two sorghum varieties.

Sorghum (Sorghum bicolor L. Moench) has been sparsely used as human food due to certain anti-nutritional factors such as tannins that reduce its digestibility, although the grain is an important source of bioactive compounds such as phenolic compounds (PCs). This study aimed to assess the impact of cooking and alkaline cooking (nixtamalization) on the bioaccessibility and antioxidant capacity of PCs of two sorghum varieties (white/red). Nixtamalization was the most effective procedure for the reduction of tannins (74.3%). Gallic acid proved to be the most bioaccessible PC (6359 µg/g). The total phenolics and condensed tannins correlated with the antioxidant capacity (ABTS/DPPH; R2: 0.30-0.43, p < 0.05). These results confirm the potential of thermal procedures to significantly modify the bioaccessibility of sorghum compounds, enhancing their concentrations and reducing anti-nutritional factors (tannins) while improving their antioxidant capacity.

Robust control of uncertain feedback linearizable systems based on adaptive disturbance estimation.

In this paper, an adaptive disturbance estimation-based control of a class of uncertain feedback linearizable systems with the presence of, both, external perturbations as well as non-modeled dynamics is considered. The aim of the control design was to solve the tracking trajectory problem for a class of output-based linearizable uncertain systems. An adaptive scheme is proposed for developing a state estimator of the uncertain dynamics. The estimation of both, the states and the uncertain dynamics is attained despite the limited knowledge of the plant and the information contained in the output signal. The uncertain section in the linearized system was approximated by a class of time-dependent combination of the system states. The observer implemented a parametric identifier to obtain the time varying parameters associated to the estimation of the uncertain section. This method ensured the adequate estimation process of the uncertainties/perturbations, measured in terms of the mean square error. Simultaneously, an adaptive gain associated to the observer adjusts its trajectories to provide the ultimate boundedness of the estimation error. Once the states of the uncertain system are obtained, a feedback controller rejects actively the perturbations that affect the system by a compensation scheme. Two numerical examples were developed to show the observer-based control performance.

Synthesis of biochar from chili seeds and its application to remove ibuprofen from water. Equilibrium and 3D modeling.

In this work chili seeds (Capsicum annuum) were used as raw material in the synthesis of biochar at temperatures between 400 and 600 °C. The samples were chemically, texturally and morphologically characterized and their properties were correlated with the calcination temperature. The adsorption mechanism of IBP was elucidated by analyzing the effect of solution pH, ionic strength and temperature, whereas that, the intraparticle diffusion mechanism was clarified through the application of a 3D diffusional model. The results evidenced that raising the pyrolysis temperature promotes a greater content of disordered graphitic carbon (51.6-85.02%) with small surface area (0.52-0.18 m2/g) and low quantity of functional groups. The adsorption study demonstrated that the biochar synthesized at 600 °C (C600) enhances the adsorption capacity >50 folds compared with chili seeds. Moreover, at pH = 7 the adsorption mechanism is governed by π-acceptor and attractive electrostatic interactions, whereas at basic pH the main adsorption mechanism is π-acceptor. Additionally, hydrophobic interactions become important by increasing the presence of NaCl. The application of 3D diffusional model based on surface diffusion interpreted clearly the kinetic curves obtaining values of Ds ranging from 2.31 × 10-8-2.51 × 10-8 cm2 s-1. Besides, it was determined that intraparticle mass flux is larger along the shortest axis of the seed, and always directed toward the particle center. The maximum mass flux takes place in the center of particle, and it advances like a moving front as time was increased.

'An investigation into the occurrence and removal of pharmaceuticals in Colombian wastewater'.

In this work, the presence of 20 pharmaceuticals in wastewater from Colombia is investigated. Several widely consumed compounds have been detected in wastewater samples from different origins and geographical areas in Colombia. The studied pharmaceuticals included antibiotics, analgesics and anti-inflammatories, cholesterol lowering statin drugs, lipid regulators, and anti-depressants. The investigated samples were urban wastewater collected during one whole week before (influent) and after treatment (effluent) in the wastewater treatment plants (WWTPs) of Bogotá and Medellin. Raw wastewater from the Hospital of Tumaco and from the city of Florencia were also collected. Analyses performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed that most of the target analytes were present in all the wastewater samples. The highest concentrations (up to 50 µg/L) corresponded to acetaminophen, but several antibiotics, such as azithromycin, ciprofloxacin and norfloxacin, and antihypertensive drugs, such as losartan and valsartan, were commonly present in influent wastewater (IWW) at levels above 1 µg/L. Moreover, the treatment applied in WWTPs seemed to not efficiently remove the compounds under study, because most pharmaceuticals were also present in effluent wastewater (EWW) at concentrations close to those of the IWW. Special emphasis was made in this work on the quality of data reported, performing a detailed study of quality control (QC) samples. The analytical approach used -direct injection of 5-fold diluted samples without any additional treatment - is simpler and faster than the commonly applied solid phase extraction (SPE). The use of 12 isotope-labelled internal standards ensured the satisfactory correction of matrix effects for the corresponding analytes. For the remaining 8 compounds, no drastic matrix effects were observed, and only four compounds (cloxacillin, doxycycline, losartan, tetracycline) presented QC recoveries near or slightly below 60%, revealing ionization suppression, particularly in the IWW. Data on the occurrence of pharmaceuticals reported in this paper are the basis for current studies that aim to develop efficient systems for the degradation/removal of these compounds from the aquatic environment.

A fuzzy approach for feature extraction of brain tissues in Non-Contrast CT / Un enfoque difuso para la extracción de características de tejidos cerebrales en TC no contrastada

Abstract: In neuroimaging, brain tissue segmentation is a fundamental part of the techniques that seek to automate the detection of pathologies, the quantification of tissues or the evaluation of the progress of a treatment. Because of its wide availability, lower cost than other imaging techniques, fast execution and proven efficacy, Non-contrast Cerebral Computerized Tomography (NCCT) is the most used technique in emergency room for neuroradiology examination, however, most research on brain segmentation focuses on MRI due to the inherent difficulty of brain tissue segmentation in NCCT. In this work, three brain tissues were characterized: white matter, gray matter and cerebrospinal fluid in NCCT images. Feature extraction of these structures was made based on the radiological attenuation index denoted by the Hounsfield Units using fuzzy logic techniques. We evaluated the classification of each tissue in NCCT images and quantified the feature extraction technique in synthetic images from real tissues with a sensitivity of 92% and a specificity of 96% for images from cases with slice thickness of 1 mm, and 96% and 98% respectively for those of 1.5 mm, demonstrating the ability of the method as feature extractor of brain tissues.

Resumen: En neuroimagen, la segmentación de tejidos cerebrales es una parte fundamental de las técnicas que buscan automatizar la detección de patologías, la cuantificación de tejidos o la evaluación del progreso de un tratamiento. Debido a su amplia disponibilidad, menor costo que otras técnicas de imagen, rápida ejecución y eficacia probada, la tomografía computarizada cerebral sin contraste (TCNC) es la técnica mayormente utilizada en emergencias para el examen neurorradiológico, sin embargo, la dificultad inherente que representa la segmentación de los tejidos cerebrales, hace que la mayoría de las investigaciones sobre la segmentación del cerebro se centren en la resonancia magnética. En este trabajo se realizó la caracterización de tres tejidos cerebrales: sustancia blanca, sustancia gris y líquido cefalorraquídeo en imágenes TCNC. Dichas estructuras fueron caracterizadas con base en el índice de atenuación radiológica denotadas por las Unidades Hounsfield utilizando técnicas de lógica difusa. Se evaluó la caracterización de cada tejido en diversos cortes de TCNC y se cuantificó la técnica de extracción de características en imágenes sintéticas a partir de tejidos reales con una sensibilidad de 92% y una especificidad de 96% para tejidos en cortes de 1 mm de grosor y 96% y 98% para los de 1.5 mm demostrando la habilidad del método como extractor de características de los tejidos cerebrales.

Lack of adherence to SHEA-IDSA treatment guidelines for Clostridium difficile infection is associated with increased mortality.

OBJECTIVES: The objective of this study was to determine our institution's compliance with 2010 Society for Healthcare Epidemiology of America and IDSA Clostridium difficile infection (CDI) treatment guidelines and their respective outcomes. METHODS: We collected clinical parameters, laboratory values, antibiotic therapy and clinical outcomes from the electronic medical records for all patients hospitalized at our institution with a diagnosis of CDI from December 2012 to November 2013. We specifically evaluated whether SHEA-IDSA treatment guidelines were followed and evaluated the associations between guideline adherence and severe outcomes including mortality. RESULTS: We identified 230 patients with CDI meeting inclusion criteria during the study period. Of these, 124 (54%) were appropriately treated, 46 (20%) were under-treated and 60 (26%) were over-treated. All-cause 90 day mortality was 17.4% overall; 43.5% in the under-treated group versus 12.9% in those appropriately treated (P < 0.0001) and 10.9% in those appropriately treated plus over-treated (P < 0.0001). Similarly, 90 day mortality attributed to CDI was 21.7% in those under-treated versus 8.9% in those appropriately treated (P = 0.03) and 8.2% in those either appropriately treated or over-treated (P = 0.015). Severe-complicated CDI occurred in 46 patients. In this subgroup, there was a non-significant trend towards increased mortality in under-treated patients (56.7%) compared with appropriately treated patients (37.5%, P = 0.35). Under-treatment was also associated with a higher rate of CDI-related ICU transfer (17.4% versus 4.8% in those appropriately treated, P = 0.023). CONCLUSIONS: Adherence to CDI treatment guidelines is associated with improved outcomes especially in those with severe disease. Increased emphasis on provision of appropriate, guideline-based CDI treatment appears warranted.

Transcriptomic and genetic studies identify NFAT5 as a candidate gene for cocaine dependence.

Cocaine reward and reinforcing effects are mediated mainly by dopaminergic neurotransmission. In this study, we aimed at evaluating gene expression changes induced by acute cocaine exposure on SH-SY5Y-differentiated cells, which have been widely used as a dopaminergic neuronal model. Expression changes and a concomitant increase in neuronal activity were observed after a 5 µM cocaine exposure, whereas no changes in gene expression or in neuronal activity took place at 1 µM cocaine. Changes in gene expression were identified in a total of 756 genes, mainly related to regulation of transcription and gene expression, cell cycle, adhesion and cell projection, as well as mitogen-activeated protein kinase (MAPK), CREB, neurotrophin and neuregulin signaling pathways. Some genes displaying altered expression were subsequently targeted with predicted functional single-nucleotide polymorphisms (SNPs) in a case-control association study in a sample of 806 cocaine-dependent patients and 817 controls. This study highlighted associations between cocaine dependence and five SNPs predicted to alter microRNA binding at the 3'-untranslated region of the NFAT5 gene. The association of SNP rs1437134 with cocaine dependence survived the Bonferroni correction for multiple testing. A functional effect was confirmed for this variant by a luciferase reporter assay, with lower expression observed for the rs1437134G allele, which was more pronounced in the presence of hsa-miR-509. However, brain volumes in regions of relevance to addiction, as assessed with magnetic resonance imaging, did not correlate with NFAT5 variation. These results suggest that the NFAT5 gene, which is upregulated a few hours after cocaine exposure, may be involved in the genetic predisposition to cocaine dependence.

LRP receptor family member associated bone disease.

A dozen years ago the identification of causal mutations in the low-density lipoprotein receptor-related protein 5 (LRP5) gene involved in two rare bone disorders propelled research in the bone field in totally new directions. Since then, there have been an explosion in the number of reports that highlight the role of the Wnt/ß-catenin pathway in the regulation of bone homeostasis. In this review we discuss some of the most recent reports (in the past 2 years) highlighting the involvement of the members of the LRP family (LRP5, LRP6, LRP4, and more recently LRP8) in the maintenance of bone and their implications in bone diseases. These reports include records of new single nucleotides polymorphisms (SNPs) and haplotypes that suggest variants in these genes can contribute to subtle variation in bone traits to mutations that give rise to extreme bone phenotypes. All of these serve to further support and reinforce the importance of this tightly regulated pathway in bone. Furthermore, we discuss provocative reports suggesting novel approaches through inhibitors of this pathway to treat rarer diseases such as Osteoporosis-Pseudoglioma Syndrome (OPPG), Osteogenesis Imperfecta (OI), and Sclerosteosis/Van Buchem disease. It is hoped that by understanding the role of each component of the pathway and their involvement in bone diseases that this knowledge will allow us to develop new, more effective therapeutic approaches for more common diseases such as post-menopausal osteoporosis, osteoarthritis, and rheumatoid arthritis as well as these rarer bone diseases.

In vivo mechanical loading rapidly activates ß-catenin signaling in osteocytes through a prostaglandin mediated mechanism.

The response of the skeleton to loading appears to be mediated through the activation of the Wnt/ß-catenin signaling pathway and osteocytes have long been postulated to be the primary mechanosensory cells in bone. To examine the kinetics of the mechanoresponse of bone and cell types involved in vivo, we performed forearm loading of 17-week-old female TOPGAL mice. ß-catenin signaling was observed only in embedded osteocytes, not osteoblasts, at 1h post-loading, spreading to additional osteocytes and finally to cells on the bone surface by 24h. This early activation at 1h appeared to be independent of receptor (Lrp5/6) mediated activation as it occurred in the presence of the inhibitors sclerostin and/or Dkk1. The COX-2 inhibitor, Carprofen, blocked the activation of ß-catenin signaling and decline in sclerostin positive osteocytes post-loading implying an important role for prostaglandin. In vitro, PI3K/Akt activation was shown to be required for ß-catenin nuclear translocation downstream from prostaglandin in MLO-Y4 osteocyte-like cells supporting this mechanism. Downstream targets of ß-catenin signaling, sclerostin and Dkk1, were also examined and found to be significantly downregulated in osteocytes in vivo at 24h post-loading. The pattern of initially activated osteocytes appeared random and in order to understand this heterogeneous expression, a novel finite element model of the strain field in the ulna was developed, which predicts highly variable local magnitudes of strain experienced by osteocytes. In summary, both in vivo and in vitro models show the rapid activation of ß-catenin in response to load through the early release of prostaglandin and that strain fields in the bone are extremely heterogeneous resulting in heterogeneous activation of the ß-catenin pathway in osteocytes in vivo.

Impact of Epstein Barr virus-related complications after high-risk allo-SCT in the era of pre-emptive rituximab.

We monitored 133 high-risk allo-SCT recipients for 6 months after transplant for EBV reactivation by quantitative real-time PCR. Rituximab was given as pre-emptive therapy for viremia >1000 copies/mL. The 1-year cumulative incidence of EBV reactivation was 29.4% (95% confidence interval (CI): 18-40) in patients monitored due to initial high-risk characteristics (n=93) and 31.8% (95% CI: 19.7-44) in those followed because of the development of refractory GVHD (n=40). Overall response rate to Rituximab was 83%. Nine patients (9.6%) developed post-transplant lymphoproliferative disorder (PTLD) at a median of +62 days after SCT. Eight of them showed a concomitant CMV reactivation. Second SCT was the only risk factor associated with EBV infection and PTLD in multivariate analysis (hazard ratio (HR) 2.6 (95% CI: 1.1-6.4; P=0.04) and HR 6.4 (95%CI: 1.3-32; P=0.02)). The development of EBV reactivation was not associated with non-relapse mortality or OS (P=0.97 and P=0.84, respectively).