Nanotechnology in reproduction: Vitamin E nanoemulsions for reducing oxidative stress in sperm cells.

Vitamin E is considered a powerful biological antioxidant; however, its characteristics such as high hydrophobicity and low stability limit its application. We propose to use nanotechnology as an innovative tool in spermatology, formulating nanoemulsions (NE) that accommodate vitamin E, protecting it from oxidation and promoting its release into the medium. The protective effect of the NE against oxidative stress was assessed in red deer epididymal sperm incubated at 37 °C. Cryopreserved sperm from eleven stags were thawed and extended to 400 × 106 sperm/ml in Bovine Gamete Medium (BGM). Once aliquoted, the samples were supplemented with the NE at different concentrations (0, 6 and 12 mM), with or without induced oxidative stress (100 µM Fe2+/ascorbate). The samples were evaluated after 0, 2 and 4 h of incubation at 37 °C. Motility (CASA), viability, mitochondrial membrane potential, acrosomal status, lipoperoxidation (C11 BODIPY 581/591), intracellular reactive oxygen species (ROS) production and DNA status (SCSA®) were assessed. After 2 and 4 h of incubation, the NE were able to prevent the deleterious effects of oxidative stress, thus improving total and progression motility (P ˂0.05). Moreover, the highest concentration tested (12 mM) improved almost every sperm kinematic variable (P ˂0.05) and preserved sperm viability in samples subjected to oxidative stress. In addition, 12 mM of NE protected the acrosomes integrity, maintained and protected mitochondrial activity, prevented sperm lipoperoxidation and reduced ROS production (P ˂0.05) in samples subjected to oxidative stress. This work indicates for the first time that vitamin E formulated in NE could be a new approach against sperm oxidative damage. This could be highly relevant for sperm physiology preservation in the context of assisted reproduction techniques.

Metals and low dose IR: Molecular effects of combined exposures using HepG2 cells as a biological model.

Uranium mining sites produce residues rich in metals and radionuclides, that may contaminate all environmental matrices, exposing human and non-human biota to low doses of ionizing radiation (LDIR) and to the chemical toxicity of several metals. To date, experimental and radio-epidemiological studies do not provide conclusive evidence of LDIR induced cancer. However, co-exposures (LDIR plus other contaminants), may increase the risks. To determine the potential for genotoxic effects in human cells induced by the exposure to LDIR plus metals, HEPG2 cells were exposed to different concentrations of a uranium mine effluent for 96 h. DNA damage was evaluated using the comet assay and changes in the expression of tumor suppressor and oncogenes were determined using qPCR. Results show that effluent concentrations higher than 5%, induce significant DNA damage. Also, a significant under-expression of ATM and TP53 genes and a significant overexpression of GADD45a gene was observed. Results show that the exposure to complex mixtures cannot be disregarded, as effects were detected at very low doses. This study highlights the need for further studies to clarify the risks of exposure to LDIR along with other stressors, to fully review the IR exposure risk limits established for human and non-human biota.

Bioprospection of Isochrysis galbana and its potential as a nutraceutical.

Microalgae have strong potential as novel sources of nutraceuticals, as they contain significant amounts of highly valuable bioactive compounds. This study focuses on the bioprospection of biomass of the microalga Isochrysis galbana and its extracts (aqueous and ethanolic), determining total polyphenols, laminarin, fucoxanthin, coenzyme Q10, and ß-carotene contents, and also assessing several biological activities (antioxidant, cytotoxicity, and hypocholesterolemic). I. galbana exhibited high phenolic content, both in aqueous and ethanolic extracts. The microalgal freeze-dried biomass presented a low laminarin content and higher content of fucoxanthin (6.10 mg per g dw), and relevant ß-carotene and Coenzyme Q10 contents were detected. I. galbana aqueous extracts presented a high antioxidant capacity (approximately 90% inhibition by the ABTS method). Furthermore, I. galbana biomass and ethanolic extract showed significant cytotoxicity against HeLa human cervical cancer cells, with IC50 values of 0.32 and 0.28 mg ml-1 respectively, demonstrating potential for further anticancer studies. The aqueous extract of I. galbana induced a significant decrease of cholesterol absorption through Caco-2 monolayers, modelling the human intestinal barrier, which suggests that it may contribute to decreasing the dietary cholesterol absorption.

Gastroprotective and anti-inflammatory activities integrated to chemical composition of Myracrodruon urundeuva Allemão - A conservationist proposal for the species.

ETHNOPHARMACOLOGICAL RELEVANCE: Myracrodruon urundeuva Allemão (Aroeira-do-Sertão), Anacardiaceae, is one of the most used plants in folk medicine in Northeastern Brazil as an anti-inflammatory, healing and antiulcer. This species is threatened with extinction due to anthropogenic exploitation. The importance of this study is to demonstrate the feasibility of a conservationist model of replacement of the M. urundeuva adult tree (inner bark) for its under developing plants (shoots) in order to ensure the preservation of this species, but also to ensure sufficient raw material for pharmaceutical purposes. AIM OF THE STUDY: To characterize chemically and assess the gastroprotective and anti-inflammatory activities of the fluid extracts from M. urundeuva innebark (adult plant) as well as stem and leaves of shoots (young plant). MATERIALS AND METHODS: The fluid extracts were prepared by maceration-percolation with hydroalcoholic solution according to the methodology described in the Brazilian Pharmacopoeia. These extracts were cleaned-up through solid phase extraction (SPE) and chemically characterized by ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-ESI-QTOF MS/MS). Gastroprotective and anti-inflammatory activities of the extracts (700 or 1000 mg/kg) were assessed on ethanol-induced gastric lesions and Croton oil-induced ear edema in rats, respectively. The extracts were evaluated for cytotoxicity in vitro. RESULTS: The UPLC-ESI-QTOF-MS/MS analysis evidenced the presence of chalcones, flavonoids and tannins. Gastroprotective and anti-inflammatory activities achieved with fluid extracts from the stems and leaves was similar to inner bark. The fluid extracts were not toxic. CONCLUSION: It is possible to replace the inner bark of the adult tree for the stems and leaves from the shoots as raw material to be used in the preparation of its the phytotherapeutics. Therefore, this finding may help in the implementation of public policies that ensure the conservation of the species along with its sustainable use for pharmaceutical purposes.

Conocimiento local del género Arceuthobium en un área natural protegida del centro de México / Local knowledge of Arceuthobium genus in a natural protected area of the centre of Mexico

El género Arceuthobium (muérdago enano) es una de las principales plagas en los bosques templados de México, con antecedentes de uso medicinal y forrajero. El objetivo de este trabajo fue documentar el conocimiento local sobre los usos del muérdago enano en el Área Natural Protegida Nevado de Toluca, México, bajo las perspectivas emic y etic, a través de la etnobotánica. Mediante entrevistas se identificó que ambas especies (A. vaginatum y A. globosum) son conocidas como muérdago y forman parte de la flora del bosque; sin embargo, los entrevistados desconocen los efectos negativos dentro de los ecosistemas forestales de esta plaga. Destacan los usos lúdicos, como juguete y tinta, y los medicinales para el tratamiento de afecciones respiratorias y del sistema nervioso, siendo éste el primer registro para dicho género localmente. Se requieren estudios que validen el conocimiento local para su inclusión en programas locales de manejo de plagas forestales.

Arceuthobium (dwarf mistletoe) genus is one of the main pests in temperate forests of Mexico, with records of medicinal and forage uses. The objective of this work was to document local knowledge regarding uses of dwarf mistletoe in the Natural Protected Area Nevado de Toluca, Mexico, under emic and etic perspectives, through ethnobotany. Throughout interviews it was identified that both species (A. vaginatum and A. globosum) are known as mistletoe and they are part of flora in forest, however, interviewees are not aware of negative effects within forest ecosystems of this pest. It stands out ludic uses as a toy and ink, and medicinal ones for treatment of respiratory diseases and nervous system, being this the first record for such genus locally. Studies are required to validate the local knowledge for its inclusion in local programmes for management of forest pests.

Lippia graveolens photochemopreventive effect against UVB radiation-induced skin carcinogenesis.

Lippia graveolens HBK (Mexican oregano) is a species that is regularly used as a condiment in Mexican cuisine. In traditional medicine, it is used for the treatment of respiratory and digestive illnesses, headaches, rheumatism and inflammation-related disorders. The main chemical components reported in this species include the following: terpenoids, iridoids and flavonoids. The aim of this study was to determine the potential photochemopreventive effect of the methanolic extract of Lippia graveolens (MELG) against ultraviolet B (UVB)-induced skin cancer in SKH-1 mice. The phenolic content, radical scavenger activity, penetration and genotoxicity of the MELG were also evaluated. The MELG exhibited scavenging activity against 1,1-diphenyl-2-picrylhydrazyl, superoxide and hydroxyl radicals, and it did not exhibit genotoxic activity in the micronucleus test. In addition, the MELG absorbed UVB (280nm) electromagnetic radiation. The main components detected in the plant extract were naringenin and galangin, and pinocembrin was also isolated and identified through spectroscopic analysis. The MELG demonstrated a photoprotective effect against UVB-induced cell death in Escherichia coli. In chronic challenge experiments, the MELG protected against UVB-induced skin cancer in SKH-1 mice. The MELG penetrated the skin of mice. Topical administration of the MELG protected against chronic UVB-induced damage in mouse SKH-1 skin. Our results suggest that the MELG has photochemopreventive activity and may potentially prevent photo-tumorigenesis.

Mineral requirements of hair sheep in tropical climates.

In this study, we evaluated the requirements of calcium (Ca), phosphorus (P), magnesium (Mg), sodium (Na) and potassium (K) for sheep hair growth. Experimental diets contained different levels of metabolizable energy [ME; 0.96, 1.28, 1.72, 2.18 and 2.62 Mcal/kg of dry matter, (DM), corresponding to 4.23, 5.64, 7.58, 9.61 and 11.55 MJ/kg DM]. The lambs' hair (n = 48) at 2 months of age presented an average body weight (BW) of 12.05 ± 1.81 kg. At the beginning of the experiment, eight animals were slaughtered as a reference group to estimate the initial empty body weight and body composition. Net mineral requirements (g/day) ranged from 0.73 to 0.71 g of Ca, 0.51 to 0.49 g of P, 0.026 to 0.026 g of Mg, 0.16 to 0.19 g of Na and 0.15 to 0.13 g of K for animals with a BW ranging from 15 to 30 kg and a daily gain of 100 g. The results of this study indicate that the net macromineral requirements for weight gain in Morada Nova lambs are different from the values commonly recommended by the Agricultural and Food Research Council.

Effects of different lipid sources on intake, digestibility and purine derivatives in hair lambs.

An experiment was conducted to evaluate the effects of different lipid sources on the nutrient intake, digestibility and purine derivative excretion of lambs. Thirty-five 60-day-old, male, non-castrated Santa Ines lambs with an initial average body weight (BW) of 13.00 ± 1.80 kg were used in a randomized complete block design with seven blocks and five treatments. The experimental treatments consisted of a control diet without supplemental lipids and four test diets with different lipid supplements, selected according to the degree of ruminal protection from hydrogenation: supplementation, being supplementation with whole cottonseed (WC), supplementation with cashew nut meal (CNM), supplementation with both cottonseed and cashew nut meal (WC-CNM) and supplementation with calcium salts of long-chain fatty acids (Ca-LCFA). The lambs were slaughtered after reaching 28 kg average BW for each treatment. The ether extract intake (EEI) was higher (p < 0.01) for the lipid supplemented compared to control diet lambs. Supplementation with WC decreased the digestibility of dry matter (DM), organic matter (OM), neutral detergent fibre (NDF) and total carbohydrate (TC) (p < 0.01), whereas supplementation with CNM, WC-CNM and Ca-LCFA reduced non-fibrous carbohydrate (NFC) digestibility (p < 0.01). The ether extract (EE) digestibility coefficient was higher with CNM, followed by Ca-LCFA and WC, when compared to WC-CNM and control diets. Nitrogen balance (NB) was not influenced (p > 0.05) by the different lipid sources. A lower purine derivative (PD) excretion and thus lower microbial protein supply (MPS) was observed for animals supplemented with Ca-LCFA (p < 0.01) compared to the WC-CNM and control diets. In conclusion, WC, CNM and WC-CNM supplementation did not have negative effects on MPS, although negative effects have been observed on nutrient digestibility.

Imágenes mentales utilizadas en diferentes actividades físicas y deportivas / Imagens mentais utilizados em diferentes atividades físicas e desportivas / Mental images used in different physical and sports activities

En esta investigación deseábamos saber si existía diferencia significativa entre distintas tareas que implicaban actividad física (pilates y bailes de salón) y tareas deportivas (fútbol sala y baloncesto) en el tipo de imagen que utilizaban las participantes. Para ello, seleccionamos un grupo de 200 mujeres (50 de pilates, 50 de bailes de salón, 50 de fútbol sala, y 50 de baloncesto), practicantes de la actividad física o del deporte, y les aplicamos la versión española del Object-Spatial Imagery and Verbal Questionnaire (OSIVQ). Posteriormente analizamos si existían diferencias significativas entre la actividad física y el deporte, y entre las diferentes actividades físicas (pilates y bailes de salón), y entre diferentes deportes (fútbol y baloncesto), en cada una de las escalas del OSIVQ. Encontramos que las practicantes de pilates tenían un procesamiento verbal, y las practicantes de bailes de salón, las de fútbol sala y las de baloncesto utilizaban imágenes del objeto. Cuando se analizó la actividad física (pilates y bailes de salón, conjuntamente), y la actividad deportiva (fútbol sala y baloncesto, conjuntamente) se encontró que tanto las participantes de la actividad física como las participantes de la actividad deportiva prefieren utilizar la imagen del objeto. Se ofrecen nuevas líneas de investigación (AU)

This study assessed significant differences in the types of mental imagery used by participants practicing sports (indoor football and basketball) or undertaking physical activity (pilates and ballroom dancing). The selected sample of 200 women (50 practicing pilates, 50 ballroom, 50 indoor football, and 50 basketball) was administered the Spanish version of the Object-Spatial Imagery and Verbal Questionnaire (OSIVQ). Significant differences in each of the scales of the OSIVQ between participants involved in sports or physical activity were assessed. Women practicing pilates were found to have verbal processing whereas ballroom dancers, indoor football or basketball players used object imagery. An analysis of physical activity (pilates and ballroom dancing) and sports (indoor football and basketball) revealed that all participants used object imagery. We propose new research lines (AU)

Nesta pesquisa, pretendemos saber se existe uma diferença significativa no tipo de imagens utilizadas pelos participantes em diferentes tarefas que envolvem a atividade física (pilates e dança de salão) e tarefas desportivas (futebol de salão e basquete). Para isso, selecionamos um grupo de 200 mulheres (50 de pilates, 50 de salão de festas, 50 de futebol de salão, e 50 de basquete), praticantes de atividade física ou desporto e lhes aplicamos a versão em espanhol do Object-Spatial Imagery and Verbal Questionnaire (OSIVQ). Avaliou-se se existiram diferenças significativas entre a actividade física e do desporto, e entre as diferentes atividades físicas (pilates e dança de salão), e entre os diferentes desportos (futebol de salão e basquete) para cada uma das escalas do OSIVQ. Verificamos que as praticantes de pilates apresentavam um processamento verbal, e as practicantes de dança de salão, de futebol de salão e de basquete utilizavam imagens do objeto. Quando se analisou a atividade física (pilates e dança de salão, juntamente) e desporto (futebol de salão e basquete, juntos), verificou-se que tanto as practicantes atividade física como as practicantes de atividade desportiva preferem usar a imagem do objeto. Propomos novas linhas de pesquisa (AU)

Micro-arc oxidation as a tool to develop multifunctional calcium-rich surfaces for dental implant applications.

Titanium (Ti) is commonly used in dental implant applications. Surface modification strategies are being followed in last years in order to build Ti oxide-based surfaces that can fulfill, simultaneously, the following requirements: induced cell attachment and adhesion, while providing a superior corrosion and tribocorrosion performance. In this work micro-arc oxidation (MAO) was used as a tool for the growth of a nanostructured bioactive titanium oxide layer aimed to enhance cell attachment and adhesion for dental implant applications. Characterization of the surfaces was performed, in terms of morphology, topography, chemical composition and crystalline structure. Primary human osteoblast adhesion on the developed surfaces was investigated in detail by electronic and atomic force microscopy as well as immunocytochemistry. Also an investigation on the early cytokine production was performed. Results show that a relatively thick hybrid and graded oxide layer was produced on the Ti surface, being constituted by a mixture of anatase, rutile and amorphous phases where calcium (Ca) and phosphorous (P) were incorporated. An outermost nanometric-thick amorphous oxide layer rich in Ca was present in the film. This amorphous layer, rich in Ca, improved fibroblast viability and metabolic activity as well as osteoblast adhesion. High-resolution techniques allowed to understand that osteoblasts adhered less in the crystalline-rich regions while they preferentially adhere and spread over in the Ca-rich amorphous oxide layer. Also, these surfaces induce higher amounts of IFN-γ cytokine secretion, which is known to regulate inflammatory responses, bone microarchitecture as well as cytoskeleton reorganization and cellular spreading. These surfaces are promising in the context of dental implants, since they might lead to faster osseointegration.

Melanin-concentrating hormone inputs to the nucleus accumbens originate from distinct hypothalamic sources and are apposed to GABAergic and cholinergic cells in the Long-Evans rat brain.

Melanin-concentrating hormone [MCH] is a neuropeptide that modulates several behaviors, such as feeding and reward. Because the hedonic and rewarding features of a food also influence feeding behavior, the nucleus accumbens [Acb] has been highlighted as a key area integrating these roles. Functional data confirm that MCH acts on a subdivision of the Acb; however, considering the importance of finding anatomical and neurochemical data that correlate the previously demonstrated function of MCH, we delineated this investigation based on the following points: (1) Is there a pattern of innervation by MCH fibers regarding the subregions within the Acb? (2) Specifically, which hypothalamic nuclei synthesize MCH and innervate the Acb? (3) Finally, what are the neurochemical identities of the accumbal neurons innervated by MCH inputs? We examined the MCH immunoreactivity [MCH-ir] in the Acb in rat brains using the peroxidase technique. Additionally, after injecting retrograde neuronal tracer [Fluoro-Gold® - FG®] into subdivisions of the Acb [shell or core], we mapped single- or double-labeled cells. Moreover, using a double immunoperoxidase protocol, we investigated the MCH-ir fibers for gamma-aminobutyric acid [GABA]-ir and choline acetyltransferase [ChAT]-ir cells in the shell subdivision of the Acb [AcbSh]. We found that the MCH-ir fibers preferentially innervate the medial AcbSh, particularly the septal pole. This innervation originated from the incerto-hypothalamic area [IHy], internuclear area, lateral hypothalamic area, perifornical area, periventricular nucleus and posterior hypothalamus. Moreover, the IHy has the highest relationship between double/single retrogradely labeled cells [n=5.33±0.66/16±0.93, i.e. 33.33%] in the whole hypothalamus. Furthermore, our data suggest that MCH-ir fibers are in apposition to GABAergic and cholinergic cells in the AcbSh. Therefore, we provide anatomical support to the ongoing functional studies investigating the relation among the hypothalamus, MCH transmission into the Acb and the involvement of known neuronal phenotypes within the AcbSh.

Variabilidad en la asistencia hospitalaria. El caso de la rehabilitación respiratoria en Colombia / Variability in hospital care. Pulmonary rehabilitation in Colombia

Objetivo: Describir las variaciones en la asistencia hospitalaria de los programas de rehabilitación respiratoria (RR) de pacientes con enfermedad pulmonar obstructiva crónica (EPOC) en un colectivo de fisioterapeutas colombianos. Materiales y métodos: Estudio observacional con diseño transversal en 250 profesionales de fisioterapia de Colombia. Se aplicó una encuesta para identificar las variaciones en los procedimientos, los componentes, la estructura y las características de los programas de RR, según las «directrices del manejo fisioterapéutico en paciente con enfermedad pulmonar con respiración espontánea» publicadas por la British Thoracic Society (BTS). Adicionalmente, se reunieron datos de información sociodemográfica. Resultados: La tasa de respuesta fue del 89,6% (n = 224). En el momento de la encuesta, el (92%) trabajaba en su mayor tiempo en programas de RR. Respecto a las variaciones en la práctica clínica, un elevado porcentaje de profesionales respondieron «realizar siempre» actividades con bajo nivel de evidencia (grados C y D), como técnicas de respiración diafragmática (48,7%), respiración con labios fruncidos durante el esfuerzo (62,9%) y técnicas de control de la respiración (37,5%). También se observó la inclusión de intervenciones con mayor grado de recomendación (grado A), como entrenamiento muscular periférico (42,9%), entrenamiento de músculos respiratorios (52,2%), educación y/o asesoramiento de la enfermedad (56,7%), uso de ventilación con presión positiva intermitente (31,7%), y aplicación de terapia de oxígeno, tanto en la fase aguda como domiciliaria (40,6%). Conclusiones: El estudio confirma importantes variaciones en los componentes terapéuticos asistenciales de la RR planteadas por la BTS para pacientes con EPOC

Objective: To describe variations in the management of pulmonary rehabilitation (PR) programs in patients with Chronic Obstructive Pulmonary Disease (COPD) by Colombian physiotherapists. Materials and method: We conducted a cross-sectional, observational study with 250 Colombian physiotherapists who routinely work with COPD patients from Colombia. A survey was made to identify variations in procedures, components, structure and characteristics of PR programs according to the «Guidelines for the physiotherapy management in patients with pulmonary disease with spontaneously breathing» published by British Thoracic Society Physiotherapy (BTS). Additionally, socio-demographic data were collected. Results: The response rate was 89.6% (n = 224). At the time of the survey, 92% of those surveied spent most of their time working in rehabilitation programs. Regarding variations in the clinical practice, the physiotherapists mainly answered «always perform» activities with low level of evidence (Grade C and D) such as diaphragmatic breathing techniques (48.7%), pursed-lip breathing during exercise (62.9%) and techniques of breath control (37.5%). A high percentage of professionals also included interventions with a higher degree of recommendation (Grade A) such as peripheral muscle training (42.9%), respiratory muscle training (52.2%), education or counseling disease (56.7%), use of intermittent positive pressure ventilation (31.7%), and application of oxygen (40.6%). Conclusions: The study confirms that there are important variations in therapeutic care components of the PR proposed by the BTS for patients with Chronic Obstructive Pulmonary Disease

Anti-Aspergillus activity of green coffee 5-O-caffeoyl quinic acid and its alkyl esters.

The antifungal activities of 5-O-caffeoyl quinic acid (5-CQA) and of methyl, butyl, octyl, and dodecyl esters or 5-CQA, were tested on five toxigenic moulds from the Aspergillus genus (Aspergillus flavus, Aspergillus nomius, Aspergillus ochraceus, Aspergillus parasiticus, Aspergillus westerdijkiae). These mycotoxin producers' moulds may contaminate many types of food crops throughout the food chain posing serious health hazard to animals and humans. The use of chemical methods to decrease mycotoxin producer moulds contamination on food crops in the field, during storage, and/or during processing, has been proved to be efficient. In this work, the antifungal effect of 5-CQA and a homologous series of 5-CQA esters (methyl, butyl, octyl, dodecyl), was investigated using the microdilution method and the minimum inhibitory concentrations (MIC50 and MIC80). All molecules presented antifungal activity, and two esters showed a MIC for all fungi: octyl (MIC50 ≤ 0.5-0.75 mg/mL, MIC80 = 1.0-1.5 mg/mL) and dodecyl (MIC50 = 0.75-1.25 mg/mL) chlorogenates. Dodecyl chlorogenate showed a MIC80 (1.5 mg/mL) only for A. parasiticus. The maximum percent of growth inhibition on aspergillii was observed with octyl (78.4-92.7%) and dodecyl (54.5-83.7%) chlorogenates, being octyl chlorogenate the most potent antifungal agent. It was thus concluded that lipophilization improved the antifungal properties of 5-CQA, which increased with the ester alkyl chain length, exhibiting a cut-off effect at 8 carbons. As far as we know, it is the first report demonstrating that lipophilization may improve the antifungal activity of 5-CQA on five toxigenic moulds from the Aspergillus genus. Lipophilization would be a novel way to synthesize a new kind of antifungal agents with a good therapeutic value or a potential use as preservative in food or cosmetics.

Clinical effectiveness of hyperbaric oxygen therapy for BK-virus-associated hemorrhagic cystitis after allogeneic bone marrow transplantation.

Late-onset hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation (HSCT) has been associated with BK virus (BKV). Antiviral drugs are of limited efficacy and the optimal treatment for HC has not yet been established. Hyperbaric oxygen (HBO) may benefit these patients. We, therefore, retrospectively evaluated the effectiveness of HBO therapy in 16 patients with HC after allogeneic HSCT. All 16 patients had macroscopic hematuria and BKV infection. Patients received 100% oxygen in a hyperbaric chamber at 2.1 atmospheres for 90 min, 5 days per week, with a median 13 treatments (range, 4-84). Fifteen patients (94%) showed complete resolution of hematuria. Median urinary DNA BKV titers declined after HBO (P<0.05). Patients started on HBO earlier after diagnosis of HC responded sooner (P<0.05). HBO was generally well tolerated and proved to be a reliable option for this difficult to manage condition.

Identification and isolation of full-length cDNA sequences by sequencing and analysis of expressed sequence tags from guarana (Paullinia cupana).

The current intense production of biological data, generated by sequencing techniques, has created an ever-growing volume of unanalyzed data. We reevaluated data produced by the guarana (Paullinia cupana) transcriptome sequencing project to identify cDNA clones with complete coding sequences (full-length clones) and complete sequences of genes of biotechnological interest, contributing to the knowledge of biological characteristics of this organism. We analyzed 15,490 ESTs of guarana in search of clones with complete coding regions. A total of 12,402 sequences were analyzed using BLAST, and 4697 full-length clones were identified, responsible for the production of 2297 different proteins. Eighty-four clones were identified as full-length for N-methyltransferase and 18 were sequenced in both directions to obtain the complete genome sequence, and confirm the search made in silico for full-length clones. Phylogenetic analyses were made with the complete genome sequences of three clones, which showed only 0.017% dissimilarity; these are phylogenetically close to the caffeine synthase of Theobroma cacao. The search for full-length clones allowed the identification of numerous clones that had the complete coding region, demonstrating this to be an efficient and useful tool in the process of biological data mining. The sequencing of the complete coding region of identified full-length clones corroborated the data from the in silico search, strengthening its efficiency and utility.

Iron deficiency and anaemia in bariatric surgical patients: causes, diagnosis and proper management.

Obesity-induced chronic inflammation leads to activation of the immune system that causes alterations of iron homeostasis including hypoferraemia, iron-restricted erythropoiesis, and finally mild-to-moderate anaemia. Thus, preoperative anaemia and iron deficiency are common among obese patients scheduled for bariatric surgery (BS). Assessment of patients should include a complete haematological and biochemical laboratory work-up, including measurement of iron stores, vitamin B12 and folate. In addition, gastrointestinal evaluation is recommended for most patients with iron-deficiency anaemia. On the other hand, BS is a long-lasting inflammatory stimulus in itself and entails a reduction of the gastric capacity and/or exclusion from the gastrointestinal tract which impair nutrients absorption, including dietary iron. Chronic gastrointestinal blood loss and iron-losingenteropathy may also contribute to iron deficiency after BS. Perioperative anaemia has been linked to increased postoperative morbidity and mortality and decreased quality of life after major surgery, whereas treatment of perioperative anaemia, and even haematinic deficiency without anaemia, has been shown to improve patient outcomes and quality of life. However, long-term follow-up data in regard to prevalence, severity, and causes of anaemia after BS are mostly absent. Iron supplements should be administered to patients after BS, but compliance with oral iron is no good. In addition, once iron deficiency has developed, it may prove refractory to oral treatment. In these situations, IV iron (which can circumvent the iron blockade at enterocytes and macrophages) has emerged as a safe and effective alternative for perioperative anaemia management. Monitoring should continue indefinitely even after the initial iron repletion and anaemia resolution, and maintenance IV iron treatment should be provided as required. New IV preparations, such ferric carboxymaltose, are safe, easy to use and up to 1000 mg can be given in a single session, thus providing an excellent tool to avoid or treat iron deficiency in this patient population.

Bisabolol-induced gastroprotection against acute gastric lesions: role of prostaglandins, nitric oxide, and KATP+ channels.

The effects of Matricaria recutita and alpha-bisabolol, a bioactive component from Chamomile species, were investigated against gastric damage induced by absolute ethanol (96%, 1 mL per animal) in rats. The effects of M. recutita extract and alpha-bisabolol on gastric mucosal damage were assessed by determination of changes in mean gastric lesion area. Mechanistic studies were carried out at with 100 mg=kg alpha-bisabolol. We further examined the possible participation of prostaglandins, nitric oxide, and KATP+ channels in its mechanism. M. recutita reduced gastric damage in all doses tested. Alpha-bisabolol at oral doses of 50 and 100 mg=kg markedly attenuated the gastric lesions induced by ethanol to the extent of 87% and 96%, respectively. Pretreatments with the nitric oxide antagonist N-nitro-l-arginine methyl ester (10 mg=kg, i.p.) or with indomethacin, an inhibitor of cyclooxygenase, failed to block effectively the gastroprotective effect of alpha-bisabolol. Furthermore, the alpha-bisabolol effect was significantly reduced in rats pretreated with glibenclamide, an inhibitor of KATP+ channel activation. Thus we provide evidence that alpha-bisabolol reduces the gastric damage induced by ethanol, at least in part, by the mechanism of activation of KATP+ channels.

Cell viability and prostacyclin release in cultured human umbilical vein endothelial cells.

Construction of efficient substitutes of human blood vessels is strongly dependent on the use of viable and fully functional cultured endothelial cells (ECs). However, very few reports have been published to date focused on the evaluation of cell viability of cultured ECs. In this work, we have determined cell viability, von Willebrand factor, and prostacyclin (PGI(2)) activity in primary cell cultures of human umbilical vein ECs, to identify the specific cell passage that is more appropriate for the development of artificial organs by tissue engineering. Cell viability was determined by quantification of the intracellular concentration of several ions by highly sensitive electron probe X-ray microanalysis, whereas von Willebrand was assayed by immunohistochemistry and PGI(2) release was quantified by radioimmunoassay. The results of our analyses demonstrate that the K/Na ratio was different for each cell passage (4.72 for the first passage, 4.55 for the second passage, and 7.82 for the third passage), suggesting that the highest cell viability corresponds to the third passage. In contrast, PGI(2) production was higher at the first two cell passages, with a significant decrease at the third passage (6.46 +/- 0.10, 5.98 +/- 0.08, and 1.62 +/- 0.05 ng/mL of supernatant for the first, second, and third passages, respectively), whereas von Willebrand expression was similar among the three cell passages analyzed in this work (64.12%, 66.66%, 65.93% of positive cells, respectively). These data suggest that cells corresponding to the second cell passage show the best ratio of viability to functionality and should therefore be used for tissue engineering protocols.

Volumetric and ionic regulation during the in vitro development of a corneal endothelial barrier.

Corneal endothelium is responsible for generating an ion flux between the corneal stroma and the anterior chamber of the eye that is necessary for the cornea to remain transparent. However, the ion transport regulatory mechanisms that develop during the formation of the endothelial barrier are not known. In this study, we determined the influence of cell confluence on cell volume and intracellular ionic content on the corneal endothelial cells of rabbits. Our results demonstrate that non-confluent endothelial cells display a hypertrophic volume increase, with higher intracellular contents of potassium and chlorine than those of confluent cells. In contrast, when cells reach confluence and the endothelial barrier forms, cell volume decreases and the intracellular contents of potassium and chlorine decrease. Our genetic analysis showed a higher expression of CFTR and CA2 genes in non-confluent cells, and of the gene KCNC3 in confluent cells. These results suggest that the normal ionic current that keeps the corneal stroma dehydrated and transparent is regulated by cell-cell contacts and endothelial cell confluence, and could explain why the loss of corneal endothelial cells is often associated with corneal edema and even blindness.