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Find video protocols related to scientific articles indexed in Pubmed.
[Contraception and breastfeeding. Spacing of pregnancies. Present concepts].
Ginecol Obstet Mex
PUBLISHED: 10-07-2014
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The risk of pregnancy in breastfeeding should be a concern of women. Family planning programs in the post natal period contraceptive choices offer high efficiency. Breastfeeding is a natural contraception method (LAM) as a contraceptive shield has 98% efficiency. Women should consider using an alternate contraceptive method when feeding requirements for this method to be effective are not met. Some of contraceptive alternatives in lactation include hormonal methods. According to the Medical Eligibility Criteria for Contraceptive Use WHO combined hormonal methods are contraindicated during breastfeeding, hormonal progestogen only methodo are considered only in Group 3 and Group 1 immediate postpartum after 6 weeks postpartum. There are modifications to these criteria by the CDC and the UK for the use of these hormones in early in lactation.
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Surface rearrangement of nanoscale zerovalent iron: the role of pH and its implications in the kinetics of arsenate sorption.
Environ Technol
PUBLISHED: 08-23-2014
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We consider the use of metallic iron nanoparticles for cleaning contaminated water, focusing our study in the sorption of arsenic compounds. In particular, we discuss the results of their sorption process on the surface of zerovalent iron nanoparticles (nZVI) by performing a complete characterization of the surface modifications. Using scanning electron microscopy, X-ray diffraction analysis and high-resolution transmission electron microscopy, spectroscopy diffraction measurements and elemental mapping, we typify the surface reconstruction during the sorption process of As(V) from aqueous solutions using nZVI when it goes into a crystalline parasymplesite structure. The experimental results were correlated to the Freundlich isotherm sorption where the sorption capacity is depleted by the increase in the pH from 4 to 7 and associated with the surface passivation of nZVI. These techniques confirm the dependence of the sorption of arsenic as a function of pH and describe the specific details on the modification of the surface area of the nanoparticles.
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[Contraception and breast feeding. Spacing of pregnancies. Present concepts].
Ginecol Obstet Mex
PUBLISHED: 07-15-2014
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The risk of pregnancy in breastfeeding should be a concern of women. Family planning programs in the postnatal period contraceptive choices offer high efficiency. Breastfeeding is a natural contraception method (LAM) as a contraceptive shield has 98 % efficiency. Women should consider using an alternate contraceptive method when feeding requirements for this method to be effective are not met. Some of contraceptive alternatives in lactation include hormonal methods. According to the Medical Eligibility Criteria for Contraceptive Use WHO combined hormonal methods are contraindicated during breast feeding, hormonal progestogen only method are considered only in Group 3 and Group 1 immediate postpartum after 6 weeks postpartum. There are modifications to these criteria by the CDC and the UK for the use of these hormones in early in lactation.
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Sphenoid brown tumor associated with a parathyroid carcinoma.
Case Rep Endocrinol
PUBLISHED: 06-12-2014
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Brown tumors are osteolytic bone lesions that occur as a result of persistent hyperparathyroidism. They usually appear late in the natural history of the disease and are currently very rare due to an earlier diagnosis of primary hyperparathyroidism. We present the case of a 53-year-old female with a 2-month history of bitemporal hemianopsia and diplopia. A computed tomography showed an osteolytic bone lesion that involved the sphenoid corpus and clivus. A biopsy was made and the histopathology result was consistent with a brown tumor. The aforementioned location is very infrequent to such tumors, and therefore represented a diagnostic challenge. However, in this case, its association to primary hyperparathyroidism was the clue for the diagnosis. The association of a brown tumor secondary to a parathyroid carcinoma has been seldom reported. This case represents, to our knowledge, one of the few brown tumors described in such location and the first secondary to a parathyroid carcinoma.
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Analysis of electron beam damage of exfoliated MoS? sheets and quantitative HAADF-STEM imaging.
Ultramicroscopy
PUBLISHED: 05-06-2014
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In this work we examined MoS? sheets by aberration-corrected scanning transmission electron microscopy (STEM) at three different energies: 80, 120 and 200 kV. Structural damage of the MoS? sheets has been controlled at 80 kV according a theoretical calculation based on the inelastic scattering of the electrons involved in the interaction electron-matter. The threshold energy for the MoS? material has been found and experimentally verified in the microscope. At energies higher than the energy threshold we show surface and edge defects produced by the electron beam irradiation. Quantitative analysis at atomic level in the images obtained at 80 kV has been performed using the experimental images and via STEM simulations using SICSTEM software to determine the exact number of MoS2? layers.
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Glutathione peroxidase 3 serum levels and GPX3 gene polymorphisms in subjects with metabolic syndrome.
Arch. Med. Res.
PUBLISHED: 04-28-2014
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Glutathione peroxidase 3 (GPx3) plays a main role in removing hydro- and lipoperoxides from the body. Changes in concentration and several single-nucleotide polymorphisms (SNP) at the GPX3 gene have been associated with vascular diseases, but the relationship of GPx3 with metabolic syndrome (MetS) remains unexplored. We undertook this study to determine the association of GPx3 serum levels and several GPX3 SNPs with the presence of MetS in Mexican subjects.
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Jumonji family histone demethylases in neural development.
Cell Tissue Res.
PUBLISHED: 03-10-2014
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Central nervous system (CNS) development is driven by coordinated actions of developmental signals and chromatin regulators that precisely regulate gene expression patterns. Histone methylation is a regulatory mechanism that controls transcriptional programs. In the last 10 years, several histone demethylases (HDM) have been identified as important players in neural development, and their implication in cell fate decisions is beginning to be recognized. Identification of the physiological roles of these enzymes and their molecular mechanisms of action will be necessary for completely understanding the process that ultimately generates different neural cells in the CNS. In this review, we provide an overview of the Jumonji family of HDMs involved in neurodevelopment, and we discuss their roles during neural fate establishment and neuronal differentiation.
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An increase in MECP2 dosage impairs neural tube formation.
Neurobiol. Dis.
PUBLISHED: 02-25-2014
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Epigenetic mechanisms are fundamental for shaping the activity of the central nervous system (CNS). Methyl-CpG binding protein 2 (MECP2) acts as a bridge between methylated DNA and transcriptional effectors responsible for differentiation programs in neurons. The importance of MECP2 dosage in CNS is evident in Rett Syndrome and MECP2 duplication syndrome, which are neurodevelopmental diseases caused by loss-of-function mutations or duplication of the MECP2 gene, respectively. Although many studies have been performed on Rett syndrome models, little is known about the effects of an increase in MECP2 dosage. Herein, we demonstrate that MECP2 overexpression affects neural tube formation, leading to a decrease in neuroblast proliferation in the neural tube ventricular zone. Furthermore, an increase in MECP2 dose provokes premature differentiation of neural precursors accompanied by greater cell death, resulting in a loss of neuronal populations. Overall, our data indicate that correct MECP2 expression levels are required for proper nervous system development.
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Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients.
Breast Cancer Res.
PUBLISHED: 02-10-2014
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Circulating tumor cells (CTCs) are commonly isolated from the blood by targeting the epithelial cell adhesion molecule (EpCAM) through positive selection. However, EpCAM can be downregulated during metastatic progression, or it can be initially not present. We designed the present prospective trial to characterize CTCs as well as other circulating cell populations in blood samples from women with metastatic breast cancer without EpCAM-dependent enrichment and/or isolation technology.
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Subthalamic nucleus lesion improves cell survival and functional recovery following dopaminergic cell transplantation in parkinsonian rats.
Eur. J. Neurosci.
PUBLISHED: 01-31-2014
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Subthalamic nucleus (STN) modulation is currently the gold standard in the treatment of Parkinson's disease (PD) cases refractory to medication. Cell transplantation is a tissue-restorative approach and is a promising strategy in the treatment of PD. One of the obstacles to overcome in cell therapy is the poor dopaminergic cell survival. Our experiment investigates the impact of a partial subthalamotomy prior to ventral mesencephalic (VM) embryonic cell transplantation on dopaminergic cell survival and functional outcome. Unilateral dopamine depletion was carried out in rats, via medial forebrain bundle (MFB) injection of 6-hydroxydopamine, and half of the animals went on to receive unilateral excitotoxic lesions of the STN/Zone Incerta (ZI) causing partial lesion of these structures on the same side as the MFB lesion. All MFB-lesioned animals, with or without the STN/ZI lesion, received striatal ipsilateral embryonic VM cell grafts. The data suggest that the STN/ZI lesion could boost the dopamine cell survival in the grafts by 2.6-fold compared with the control grafted-only group. Moreover, performance on the drug-induced rotation and the spontaneous behavior tests were ameliorated on the STN/ZI-lesioned group to a significantly greater extent than the grafted-only group. These data suggest that the STN/ZI partial lesion optimized the striatal environment, promoting an improvement in cell survival. Further studies are needed to see whether the synergy between STN modulation via deep brain stimulation and cell therapy might have clinical applications in the management of PD.
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Melatonin content of pepper and tomato fruits: effects of cultivar and solar radiation.
Food Chem
PUBLISHED: 01-30-2014
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We evaluated the effect of cultivar and solar radiation on the melatonin content of Capsicum annuum (pepper) and Solanum lycopersicum (tomato) fruits. The melatonin content of red pepper fruits ranged from 31 to 93ngg(-1) (dry weight). The melatonin content of tomato ranged from 7.5 to 250ngg(-1) (dry weight). We also studied the effect of ripeness on melatonin content and identified one group of pepper cultivars in which the melatonin content increased as the fruit ripened and another in which it decreased as the fruit ripened. Under shade conditions, the melatonin content in most of tomato cultivars tended to increase (up to 135%), whereas that of most pepper cultivars decreased (to 64%). Overall, the results also demonstrated that the melatonin content of the fruits was not related to carbon fluxes from leaves.
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Proteomic analyses of baculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus budded and occluded virus.
J. Gen. Virol.
PUBLISHED: 01-17-2014
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Baculoviruses infect insects, producing two distinct phenotypes during the viral life cycle: the budded virus (BV) and the occlusion-derived virus (ODV) for intra- and inter-host spread, respectively. Since the 1980s, several countries have been using Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) as a biological control agent against the velvet bean caterpillar, A. gemmatalis. The genome of AgMNPV isolate 2D (AgMNPV-2D) carries at least 152 potential genes, with 24 that possibly code for structural proteins. Proteomic studies have been carried out on a few baculoviruses, with six ODV and two BV proteomes completed so far. Moreover, there are limited data on virion proteins carried by AgMNPV-2D. Therefore, structural proteins of AgMNPV-2D were analysed by MALDI- quadrupole-TOF and liquid chromatography MS/MS. A total of 44 proteins were associated with the ODV and 33 with the BV of AgMNPV-2D. Although 38 structural proteins were already known, we found six new proteins in the ODV and seven new proteins carried by the AgMNPV-2D BV. Eleven cellular proteins that were found on several other enveloped viruses were also identified, which are possibly carried with the virion. These findings may provide novel insights into baculovirus biology and their host interaction. Moreover, our data may be helpful in subsequent applied studies aiming to improve AgMNPV use as a biopesticide and a biotechnology tool for gene expression or delivery.
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Molecular characterization of Mycobacterium bovis isolates from patients with tuberculosis in Baja California, Mexico.
Infect. Genet. Evol.
PUBLISHED: 01-07-2014
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The incidence of tuberculosis (TB) from Mycobacterium bovis in humans is likely to be underestimated and in some cases even ignored in most developing countries. This may be due to the difficulty of differentiating TB caused by either Mycobacteriumtuberculosis or M. bovis. Our objectives were to determine the prevalence of M. bovis human disease among the patients referred for study to the Tuberculosis Laboratory of the Tijuana General Hospital in Baja California, Mexico and to characterize molecularly the clinical isolates using 8 loci of MIRU-VNTR. A cross-sectional analysis of all culture-proven cases of tuberculosis was conducted during the period from January 1, 2011 through June 30, 2013. Clinical isolates that exhibited resistance to pyrazinamide (Z) were submitted for molecular analysis. A total of 2699 clinical samples were cultured during the study period and 600 (22%) that tested positive were processed for drug susceptibility for first line drugs. Sixty-four (10.7%) of the tested isolates tested were resistant to Z, and 27 (4.5%) of those were subsequently identified molecularly as M. bovis. Three of the M. bovis isolates were polyresistant to Z, isoniazid (H), ethambutol (E) and rifampicin (R) (Z+H+E, Z+E and Z+R); the rest were only resistant only to Z. VNTR typing, based on the 8 VNTR loci commonly tested for M.bovis, detected 12 allelic profiles (genotypes). The real burden of M. bovis cases among the total reported human tuberculosis cases can only be known from especially designed studies in which, during a specific period, all specimens submitted to tuberculosis diagnosis in one or more laboratories are cultured on the appropriate media and the isolated mycobacteria are analyzed to differentiate M. bovis from M. tuberculosis and other Mycobacterium species.
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Ryanodine receptor-mediated Ca(2+) release underlies iron-induced mitochondrial fission and stimulates mitochondrial Ca(2+) uptake in primary hippocampal neurons.
Front Mol Neurosci
PUBLISHED: 01-01-2014
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Mounting evidence indicates that iron accumulation impairs brain function. We have reported previously that addition of sub-lethal concentrations of iron to primary hippocampal neurons produces Ca(2) (+) signals and promotes cytoplasmic generation of reactive oxygen species. These Ca(2) (+) signals, which emerge within seconds after iron addition, arise mostly from Ca(2) (+) release through the redox-sensitive ryanodine receptor (RyR) channels present in the endoplasmic reticulum. We have reported also that addition of synaptotoxic amyloid-? oligomers to primary hippocampal neurons stimulates RyR-mediated Ca(2) (+) release, generating long-lasting Ca(2) (+) signals that activate Ca(2) (+)-sensitive cellular effectors and promote the disruption of the mitochondrial network. Here, we describe that 24 h incubation of primary hippocampal neurons with iron enhanced agonist-induced RyR-mediated Ca(2) (+) release and promoted mitochondrial network fragmentation in 43% of neurons, a response significantly prevented by RyR inhibition and by the antioxidant agent N-acetyl-L-cysteine. Stimulation of RyR-mediated Ca(2) (+) release by a RyR agonist promoted mitochondrial Ca(2) (+) uptake in control neurons and in iron-treated neurons that displayed non-fragmented mitochondria, but not in neurons with fragmented mitochondria. Yet, the global cytoplasmic Ca(2) (+) increase induced by the Ca(2) (+) ionophore ionomycin prompted significant mitochondrial Ca(2) (+) uptake in neurons with fragmented mitochondria, indicating that fragmentation did not prevent mitochondrial Ca(2) (+) uptake but presumably decreased the functional coupling between RyR-mediated Ca(2) (+) release and the mitochondrial Ca(2) (+) uniporter. Taken together, our results indicate that stimulation of redox-sensitive RyR-mediated Ca(2) (+) release by iron causes significant neuronal mitochondrial fragmentation, which presumably contributes to the impairment of neuronal function produced by iron accumulation.
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Association analysis between -308G/A and -238G/A TNF-alpha gene promoter polymorphisms and insulin resistance in Mexican women with gestational diabetes mellitus.
J. Investig. Med.
PUBLISHED: 12-20-2013
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Gestational diabetes mellitus (GDM) is characterized by insulin resistance. It has been described that tumor necrosis factor ? (TNF-?) plays a key role in the pathogenesis of insulin resistance; moreover, increased levels of this proinflammatory cytokine have been reported in women with GDM. Therefore, this study was aimed to assess the presence of associations between the -308G/A and -238G/A polymorphisms and specific haplotypes of the TNF-? gene promoter region and insulin resistance in Mexican women with GDM.
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Detection of respiration in central venous pressure using state machine.
Conf Proc IEEE Eng Med Biol Soc
PUBLISHED: 10-11-2013
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Reliable information from patient monitors enhances treatment for critically ill patients. Redundant sources for information would aid identification of faulty sensors and leads, and improve presentation of physiological data. Respiratory information can be obtained from several sources, including airway pressure and central venous pressure (CVP). CVP signals have been analyzed using frequency information to isolate the respiration related part of the signal or to obtain statistics about respiration. This study uses a state machine algorithm to detect the timing of each cycle of respiration. A state machine has advantages of enforcing a predictable cycle of expiration and inspiration. The detection of respiratory cycles can be done in real-time, allowing identification of irregular periods between inspirations and prolonged periods with no inspiration, for which an alert may be issued. The algorithm was tested on data obtain from the PhysioNet database of recordings from intensive care patients. The airway pressure signal was used to determine the "true values" of the timing of each respiratory cycle for checking the accuracy of the algorithm analyzing the CVP signal. Parameters of the algorithm were found that would result in a true positive value of above 98% for detection of each cycle of respiration from analysis of the CVP signal, compared to analysis of the RESP signal.
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Spectroscopy analysis of chitosan-glibenclamide hydrogels.
Spectrochim Acta A Mol Biomol Spectrosc
PUBLISHED: 07-17-2013
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The structure of glibenclamide, 5-chloro-N-(2-{4-[(cyclohexylamino)carbonyl] aminosulfonyl}phenyl) ethyl)-2-methoxybenzamide, an important antidiabetic drug, has been studied both chitosan using theoretical calculations like Gibbs free energy, electrostatic potential, FTIR and NMR spectroscopy. Fourier transform infrared (FT-IR) spectroscopy reveals information about the molecular interactions of chemical components and is useful to characterization of hydrogel. Nucleophilic and electrophilic regions were calculated using the electrostatic potential.
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In Vivo Noninvasive Imaging of Healthy Lower Lip Mucosa: A Correlation Study between High-Definition Optical Coherence Tomography, Reflectance Confocal Microscopy, and Histology.
Dermatol Res Pract
PUBLISHED: 07-04-2013
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In recent years, technology has allowed the development of new diagnostic techniques which allow real-time, in vivo, noninvasive evaluation of morphological changes in tissue. This study compares and correlates the images and findings obtained by high-definition optical coherence tomography (HD-OCT) and reflectance confocal microscopy (RCM) with histology in normal healthy oral mucosa. The healthy lip mucosa of ten adult volunteers was imaged with HD-OCT and RCM. Each volunteer was systematically evaluated by RCM starting in the uppermost part of the epithelium down to the lamina propia. Afterwards, volunteers were examined with a commercially available full-field HD-OCT system using both the "slice" and the "en-face" mode. A "punch" biopsy of the lower lip mucosa was obtained and prepared for conventional histology. The architectural overview offered by "slice" mode HD-OCT correlates with histologic findings at low magnification. In the superficial uppermost layers of the epithelium, RCM imaging provided greater cellular detail than histology. As we deepened into the suprabasal layers, the findings are in accordance with physiological cellular differentiation and correlate with the images obtained from conventional histology. The combined use of these two novel non-invasive imaging techniques provides morphological imaging with sufficient resolution and penetration depth, resulting in quasihistological images.
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[Mirizzi syndrome: experience at Spanish Hospital of Veracruz].
Cir Cir
PUBLISHED: 06-18-2013
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Mirizzi syndrome is a complication of gallbladder stones impacted in Hartmanns pouch or cystic duct with compression of the bile duct. The diagnosis is made by imaging studies, although most of them are found through intraoperative surgical findings. Treatment is cholecystectomy and bile duct restoration when needed.
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[Pharmacogenetics and antiepileptic drug metabolism: implication of genetic variants in cytochromes P450].
Rev Neurol
PUBLISHED: 05-01-2013
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Antiepileptic drugs (AEDs) are used for the seizures control in patients with epilepsy, however 20-30% of epileptic patients are drug resistant. Several factors contributing to the variability of the AEDs response, and this variability can be partially attributed to the presence of sequence variations (polymorphisms) in genes encoding enzymes involved in the AEDs metabolism.
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Hold on tightly: how to keep the local activation of small GTPases.
Cell Adh Migr
PUBLISHED: 04-16-2013
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Signaling regulated by Rho small GTPases plays a pivotal role in cell migration, cell attachment to substratum or to their neighbors among other functions. Concerted efforts have focused on understanding how different GTPases are activated by specific stimuli and which regulator is responsible for the spatio-temporal control of their activity at particular intracellular sites. We have recently described the role of a scaffold protein, Ajuba, in adherens junction maintenance via direct stabilization of activated small GTPase Rac1 at cell-cell contacts. Ajuba binds to both active and inactive forms of Rac1. Upon junction formation, Rac1 activation initiates a positive feedback loop leading to Ajuba phosphorylation and Ajuba-mediated retention of activated Rac1 at junctions. Thus, cytoskeletal proteins may have a dual role to provide a scaffolding platform and dynamically modulate small GTPases function at a specific place, irrespective of their ability to interact with active and inactive forms. Here we discuss similar mechanisms via which cytoskeletal proteins can facilitate cellular processes downstream of Rho proteins by increasing their affinity to activated GTPases.
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Low prevalence of interleukin-6 haplotypes associated with a decreased risk of type 2 diabetes in Mexican subjects with a family history of type 2 diabetes.
Arch. Med. Res.
PUBLISHED: 03-20-2013
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There is evidence that family history of type 2 diabetes (FHT2D) and single nucleotide polymorphisms (SNP) on the IL-6 gene promoter region are separately associated with the risk of developing type 2 diabetes. However the relationship between adult Mexican subjects with FHT2D and genotypes/haplotypes for IL-6 gene has not been explored. The aim of the present work was to study the prevalence of IL-6 -598G>A-572G>C-174G>C haplotypes among subjects with FHT2D and to determine whether their presence influences the relationship between FHT2D and risk factors for diabetes.
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Innovative continuous non-invasive cuffless blood pressure monitoring based on photoplethysmography technology.
Intensive Care Med
PUBLISHED: 03-06-2013
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To develop and validate a continuous non-invasive blood pressure (BP) monitoring system using photoplethysmography (PPG) technology through pulse oximetry (PO).
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Thermoplastic starch films reinforced with talc nanoparticles.
Carbohydr Polym
PUBLISHED: 03-04-2013
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Nanocomposite films of thermoplastic corn starch (TPS) with talc particles were obtained by thermo-compression in order to study the effect of filler on structure, optical, and thermal properties. Talc increased the films rigid phase, thus their cross-sections resulted more irregular. Talc preferential orientation within matrix and good compatibility between particles and TPS was observed by SEM. Slight crystalline structure changes in TPS matrix were measured by XRD and DSC, due to talc nucleating effect. Randomly dispersed talc nanoagglomerates and individual platelets were assessed by TEM. Laminar morphology and nano-sized particles allowed that nanocomposite films were optically transparent. TPS-talc films resulted heterogeneous materials, presenting domains rich in glycerol and others rich in starch. Talc incorporation higher than 3%, w/w increased softening resistance of the nanocomposites as stated by DMA. Relaxation temperatures of glycerol-rich phase shifted to higher values since talc reduces the mobility of starch chains.
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Use of sugammadex in Strumpell-Lorrain disease: a report of two cases.
Rev Bras Anestesiol
PUBLISHED: 02-27-2013
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Strumpell-Lorrain disease - or familial spastic paraplegia (FSP) - is a rare hereditary neurological disorder, mainly characterized by variable degrees of stiffness and weakening of the muscles, with cognitive impairment, deafness, and ataxia in the more severe cases. We describe two female siblings with FSP programmed for cholecystectomy and subtotal colectomy, respectively, and also how we dealt with the anesthetic management in both cases and review the literature on this disease in relation to anesthesia.
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[Socioeconomic inequalities in the onset and progression of disability in a cohort of older people in Santiago (Chile)].
Gac Sanit
PUBLISHED: 01-03-2013
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To describe and compare socioeconomic inequalities in the onset and progression of disability in a cohort of Chilean community-dwelling older people that provides unprecedented information about this process in Chile.
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Application of colon capsule endoscopy (CCE) to evaluate the whole gastrointestinal tract: a comparative study of single-camera and dual-camera analysis.
Clin Exp Gastroenterol
PUBLISHED: 01-01-2013
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Colon capsule endoscopy (CCE) was developed for the evaluation of colorectal pathology. In this study, our aim was to assess if a dual-camera analysis using CCE allows better evaluation of the whole gastrointestinal (GI) tract compared to a single-camera analysis.
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Identification of male gametogenesis expressed genes from the scallop Nodipecten subnodosus by suppressive subtraction hybridization and pyrosequencing.
PLoS ONE
PUBLISHED: 01-01-2013
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Despite the great advances in sequencing technologies, genomic and transcriptomic information for marine non-model species with ecological, evolutionary, and economical interest is still scarce. In this work we aimed to identify genes expressed during spermatogenesis in the functional hermaphrodite scallop Nodipecten subnodosus (Mollusca: Bivalvia: Pectinidae), with the purpose of obtaining a panel of genes that would allow for the study of differentially transcribed genes between diploid and triploid scallops in the context of meiotic arrest and reproductive sterility. Because our aim was to isolate genes involved in meiosis and other testis maturation-related processes, we generated suppressive subtractive hybridization libraries of testis vs. inactive gonad. We obtained 352 and 177 ESTs by clone sequencing, and using pyrosequencing (454-Roche) we maximized the identified ESTs to 34,276 reads. A total of 1,153 genes from the testis library had a blastx hit and GO annotation, including genes specific for meiosis, spermatogenesis, sex-differentiation, and transposable elements. Some of the identified meiosis genes function in chromosome pairing (scp2, scp3), recombination and DNA repair (dmc1, rad51, ccnb1ip1/hei10), and meiotic checkpoints (rad1, hormad1, dtl/cdt2). Gene expression analyses in different gametogenic stages in both sexual regions of the gonad of meiosis genes confirmed that the expression was specific or increased towards the maturing testis. Spermatogenesis genes included known testis-specific ones (kelch-10, shippo1, adad1), with some of these known to be associated to sterility. Sex differentiation genes included one of the most conserved genes at the bottom of the sex-determination cascade (dmrt1). Transcript from transposable elements, reverse transcriptase, and transposases in this library evidenced that transposition is an active process during spermatogenesis in N. subnodosus. In relation to the inactive library, we identified 833 transcripts with functional annotation related to activation of the transcription and translation machinery, as well as to germline control and maintenance.
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[Full naming with bi-dimensional abstract stimuli in six-year-old children].
Psicothema
PUBLISHED: 11-04-2011
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The aim of this study was to evaluate a procedure to reveal the emergence of full naming in typically developing children. In Experiment 1, five 6-year-old children (a) learned tacts of pictures and the emergence of the selection of these pictures upon hearing their names was tested; and (b) the selection of other pictures was taught and the emergence of the tacts was tested. All children showed the emergence of picture selection and 3 children showed the emergence of the tacts. In Experiment 2a, the childrens correct repetition of the names of 3 pictures was verified first, then the experimenter presented the pictures with their names without requiring any behavior other than attending. Finally, the emergence of picture selection and the tacts was tested. Experiment 2b replicated Experiment 2a with words that were easier to pronounce. Picture selection and the tacts emerged; this performance documented full naming for the first time with typically developing children. These procedures can be used to induce naming capacity in children who lack it; hence, they can be applied to teach children with learning delays.
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Systemic lupus erythematosus risk factors for coronary artery calcifications.
Rheumatology (Oxford)
PUBLISHED: 10-29-2011
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Premature atherosclerosis in patients with SLE is partially explained by traditional risk factors; therefore, we aimed to identify lupus-related risk factors for coronary artery calcifications.
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[Sirenomelia apus. Case report].
Ginecol Obstet Mex
PUBLISHED: 10-05-2011
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We report a case of Sirenomelia. The mother began prenatal care in the second trimester. Transabdominal ultrasound was determined anhydramnios, cardiac abnormalities and lumbosacral spine. We obtained a single fetus of 21 weeks gestation with fused lower extremities from the hip to finish in a stump without the presence of feet. Heart with transposition of the great vessels, among other birth defects. It was classified as symelia, Apodi apus, monopodio sirenoide, siren ectropodia, type VI. It is important to diagnose early, because it is a serious and deadly disorder.
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[Association of age with sperm DNA fragmentation].
Rev Med Chil
PUBLISHED: 08-25-2011
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There is an association between aging ana an increased number of sperms with alterations in nuclear DNA.
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Sampling scarab beetles in tropical forests: the effect of light source and night sampling periods.
J. Insect Sci.
PUBLISHED: 08-01-2011
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Light traps have been used widely to sample insect abundance and diversity, but their performance for sampling scarab beetles in tropical forests based on light source type and sampling hours throughout the night has not been evaluated. The efficiency of mercury-vapour lamps, cool white light and ultraviolet light sources in attracting Dynastinae, Melolonthinae and Rutelinae scarab beetles, and the most adequate period of the night to carry out the sampling was tested in different forest areas of Costa Rica. Our results showed that light source wavelengths and hours of sampling influenced scarab beetle catches. No significant differences were observed in trap performance between the ultraviolet light and mercury-vapour traps, whereas these two methods caught significantly more species richness and abundance than cool white light traps. Species composition also varied between methods. Large differences appear between catches in the sampling period, with the first five hours of the night being more effective than the last five hours. Because of their high efficiency and logistic advantages, we recommend ultraviolet light traps deployed during the first hours of the night as the best sampling method for biodiversity studies of those scarab beetles in tropical forests.
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Expression and activity of trypsin and pepsin during larval development of the spotted rose snapper Lutjanus guttatus.
Comp. Biochem. Physiol. B, Biochem. Mol. Biol.
PUBLISHED: 06-15-2011
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The present study aimed to describe and understand the development of the digestive system in spotted rose snapper (Lutjanus guttatus) larvae from hatching to 40 days post-hatch (dph). The mouth opened between 2 and 3 dph, at that moment the digestive tract was barely differentiated into the anterior and posterior intestine, although the liver and pancreas were already present. Gastric glands were observed until 20 dph, followed by the differentiation of the stomach between 20 and 25 dph. Trypsinogen expression and trypsin activity were detected at hatching, increasing concomitantly to larval development and the change in the type of food. Maximum levels of trypsinogen expression were observed at 25 dph, when animals were fed with Artemia nauplii, and maximum trypsin activity was detected at 35 dph, when larvae were fed with an artificial diet. On the other hand, pepsinogen gene expression was detected at 18 dph, two days before pepsin enzymatic activity and appearance of gastric glands. Maximum pepsin activity was also observed at 35 dph. These results suggest that in this species weaning could be initiated at an earlier age than is currently practiced (between 28 and 30 dph), since larvae of spotted rose snapper develop a functional stomach between days 20 and 25 post-hatch.
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Post-transcriptional gene silencing by RNA interference in non-mammalian vertebrate systems: where do we stand?
Mutat. Res.
PUBLISHED: 06-04-2011
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RNA interference (RNAi), the process by which double stranded RNA induces the silencing of endogenous genes through the degradation of its correspondent messenger RNA, has been used for post-transcriptional gene silencing allowing scientists to better understand gene function, becoming a powerful tool in reverse genetics for in vivo and in vitro systems. Successful results in vivo have been obtained from invertebrate animal models, whereas vertebrate systems have been limited primarily to mammalian models and cell lines. Nevertheless, exciting results have also been reported from non-mammalian vertebrate models, such as the knock-down of endogenous genes in Xenopus tadpoles by a construct containing both a Xenopus-specific shRNA sequence and the human Ago2 (which is a key enzyme in the RNAi silencing complex), or the design of a novel vector expressing a miRNA driven by a tissue-specific promoter in zebrafish, and the use of an avian retroviral vector to deliver miRNA and shRNA in chicken embryos proving to be effective in knocking-down endogenous genes with a long lasting effect, to mention some examples. Whether dsRNA is able to initiate a specific RNAi response, or all the factors required for RNAi are present in non-mammalian vertebrates, are still questions which remain to be answered. Further progress in understanding natural RNAi mechanisms in non-mammalian vertebrates will help scientists to overcome difficulties and improve this gene silencing technology. There is no doubt that in few years RNAi silencing approaches will become the tool of choice to knock-down genes in all groups of non-mammalian vertebrates, fulfilling different purposes, from basic research to animal therapeutics and drug discovery.
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Tumor necrosis factor-alpha gene promoter -308G/A and -238G/A polymorphisms in Mexican patients with type 2 diabetes mellitus.
Dis. Markers
PUBLISHED: 04-22-2011
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The association between some Tumor necrosis factor-alpha (TNF-?) promoter polymorphisms and Type 2 diabetes mellitus (T2DM) remains controversial. Ethnic differences may play a role in these conflicting results. The aim of this study was to investigate the association between -308G/A and -238G/A polymorphisms located in the promoter region of the TNF-? gene and T2DM in Mexican mestizo patients. Nine hundred four individuals (259 patients with T2DM and 645~controls) were genotyped for the -308G/A and -238G/A polymorphisms by PCR--RFLP. We found that the -238A allele increased the risk of developing T2DM in Mexican patients (OR=1.57, 95% CI: 1.07-2.29; p=0.018). Moreover, we found that the frequency of the GA haplotype (created by the -308G and -238A alleles) was significantly increased in patients with T2DM when compared with controls (OR =1.56, 95% CI: 1.05-2.31; p=0.026). Our results suggest that the -238G/A polymorphism and a specific haplotype (GA) are genetic risk factors for the development of T2DM in Mexican population.
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Bilateral breast carcinoma: clinical characteristics and its impact on survival.
Breast J
PUBLISHED: 11-13-2010
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The higher incidence of breast cancer, the improvements in diagnosis and treatment, together with the growing life expectancy have brought about an increase in the number of patients at risk for bilateral breast carcinoma. The aim of this study is to describe the characteristics of patients suffering from bilateral breast carcinoma who underwent surgery at the Breast Pathology Service of the Buenos Aires British Hospital and to analyze impact on survival. Between January 1970 and May 2007, 4,085 cases of breast carcinoma in 3,864 patients were treated at the Breast Diseases Division of the Buenos Aires British Hospital. A retrospective study of 194 patients with bilateral breast carcinoma was carried out: 80 synchronous and 114 metachronous. In order to compare survival, a group of 2,237 patients with unilateral breast carcinoma who had undergone surgery was analyzed. The risk of developing a contralateral breast carcinoma was 0.9% per year, with an accumulated risk at 15 years of 12.75%. The 5-year survival was 85.9% for unilateral carcinomas, 94.6% for metachronous carcinoma, and 63.3% for synchronous carcinoma. The 15-year survival was 65.5% for unilateral carcinomas, 52.3% for metachronous, and 37.2% for synchronous. The incidence of bilateral carcinomas is low. Survival was worse in patients with metachronous carcinoma diagnosed within 5 years of the first malignancy. Survival in patients with metachronous carcinoma diagnosed after 5 years is similar to those with unilateral carcinoma. Synchronous carcinoma was associated to worse survival, being an independent risk factor for mortality.
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Oncogenic activation of the Notch1 gene by deletion of its promoter in Ikaros-deficient T-ALL.
Blood
PUBLISHED: 09-09-2010
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The Notch pathway is frequently activated in T-cell acute lymphoblastic leukemias (T-ALLs). Of the Notch receptors, Notch1 is a recurrent target of gain-of-function mutations and Notch3 is expressed in all T-ALLs, but it is currently unclear how these receptors contribute to T-cell transformation in vivo. We investigated the role of Notch1 and Notch3 in T-ALL progression by a genetic approach, in mice bearing a knockdown mutation in the Ikaros gene that spontaneously develop Notch-dependent T-ALL. While deletion of Notch3 has little effect, T cell-specific deletion of floxed Notch1 promoter/exon 1 sequences significantly accelerates leukemogenesis. Notch1-deleted tumors lack surface Notch1 but express ?-secretase-cleaved intracellular Notch1 proteins. In addition, these tumors accumulate high levels of truncated Notch1 transcripts that are caused by aberrant transcription from cryptic initiation sites in the 3 part of the gene. Deletion of the floxed sequences directly reprograms the Notch1 locus to begin transcription from these 3 promoters and is accompanied by an epigenetic reorganization of the Notch1 locus that is consistent with transcriptional activation. Further, spontaneous deletion of 5 Notch1 sequences occurs in approximately 75% of Ikaros-deficient T-ALLs. These results reveal a novel mechanism for the oncogenic activation of the Notch1 gene after deletion of its main promoter.
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Single anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S). One to three-year follow-up.
Obes Surg
PUBLISHED: 08-28-2010
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Single anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S) is a new operation for morbid obesity based on the biliopancreatic diversion in which a sleeve gastrectomy is followed by an end-to-side duodeno-ileal diversion. The preservation of the pylorus makes possible the reconstruction in one loop, which reduces operating time and needs no mesentery opening. We review the results obtained on the first 50 operated patients with 1 to 3 years follow-up. Eighteen men and 32 women with a mean BMI of 44 kg/m(2) were operated on. Hypertension was present in 50%, sleep apnea in 30%, hypertriglyceridemia in 60% and hypercholesterolemia in 43%.There were 27 type two diabetics, most of them on insulin therapy. There were two gastric staple-line leaks and one long-term subphrenic abscess. Follow-up is complete for 98% of the patients. Excess weight loss reached 94.7% at 1 year, and it was maintained over the second and third year. At 1 year, mild anemia has been detected in 10% of the cases. Albumin concentration was under normal levels in 8% of the patients in the first postoperative year, but all patients recovered to normal levels by the third postoperative year. All diabetic patients have normalized glucose or HbA1c levels after the sixth postoperative month with no need of anti-diabetic therapy. SADI-S is a promising operation which offers excellent weight loss and metabolic results. The elimination of one anastomosis reduces operative time and decreases the possibility of surgically related complications.
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[Cardiovascular prevention according to CEIPC: a critical appraisal].
Rev. Esp. Salud Publica
PUBLISHED: 06-24-2010
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We present a critical appraisal of the adaptation to the 4th European Guidelines on Cardiovascular Disease Prevention in Clinical Practice carried out by The Spanish Interdisciplinary Committee for Cardiovascular Disease Prevention (CEIPC), which is based on: 1) the removal, by the CEIPC, of important restrictions on the start of drug therapy that are contained in the European guide 2) the existence of internal contradictions and differing recommendations regarding the goals of LDL in the several publications of the CEIP adaptation; and 3) and the almost total lack of necessary discussion about risk tables in Spain. Therefore, it makes a critical appraisal of some of the most important clinical recommendations shared by the CEIPC and the European guide that are not supported by clinical evidence, like the implicit proposal of using the estimated cardiovascular risk as a target for treatment, the criteria to begin the antihypertensive drug therapy and therapeutic goals for blood pressure, LDL-cholesterol and HbA1c. The public health administration and also the scientific society must ensure transparency and independence in the drafting of documents endorsed by them, including the management and declaration of potential conflicts of interest among editors and group members. The public health administration and also the scientific society must guarantee a framework of honesty and transparency in the documents endorsed by them, with a complete declaration of the authors conflict of interests.
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Influence of CRP, IL6, and TNFA gene polymorphisms on circulating levels of C-reactive protein in Mexican adolescents.
Arch. Med. Res.
PUBLISHED: 05-31-2010
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Obesity correlates with a chronic and low-grade inflammation status. C-reactive protein (CRP) measurement has been used as an independent risk marker for future cardiovascular events. CRP level shows interindividual variability due to environmental and genetic factors. The aim of this study was to assess the association of functional polymorphisms on CRP, IL6, and TNFA genes with serum CRP levels in Mexican mestizo adolescents.
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Activation of geminivirus V-sense promoters in roots is restricted to nematode feeding sites.
Mol. Plant Pathol.
PUBLISHED: 05-08-2010
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Obligate sedentary endoparasitic nematodes, such as the root-knot and cyst nematodes, elicit the differentiation of specialized nematode nurse or feeding cells [nematode feeding sites (NFS), giant cells and syncytia, respectively]. During NFS differentiation, marked changes in cell cycle progression occur, partly similar to those induced by some geminiviruses. In this work, we describe the activation of V-sense promoters from the Maize streak virus (MSV) and Wheat dwarf virus (WDV) in NFS formed by root-knot and cyst nematodes. Both promoters were transiently active in microinjection experiments. In tobacco and Arabidopsis transgenic lines carrying promoter-beta-glucuronidase fusions, the MSV V-sense promoter was activated in the vascular tissues of aerial plant parts, primarily leaf and cotyledon phloem tissue and some floral structures. Interestingly, in roots, promoter activation was restricted to syncytia and giant cells tested with four different nematode populations, but undetectable in the rest of the root system. As the activity of the promoter in transgenic rootstocks should be restricted to NFS only, the MSV promoter may have utility in engineering grafted crops for nematode control. Therefore, this study represents a step in the provision of some of the much needed additional data on promoters with restricted activation in NFS useful in biotechnological nematode control strategies.
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Heat-shock protein (Hsp70) and cytochrome P-450 (CYP1A) in the white mullet Mugil curema (Pisces:Mugilidae) as biomarkers to assess environmental quality in coastal lagoons.
J Environ Sci Health A Tox Hazard Subst Environ Eng
PUBLISHED: 04-15-2010
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Biomarkers have been useful tools to monitor some effects of pollution in coastal environments. Hepatic expression of heat-shock protein 70 (Hsp70) and cytochrome P450 1A (CYP1A) were analyzed in white mullet (Mugil curema) by RT-PCR from July, 2005 until July, 2006 in three coastal lagoons located in the southern Gulf of California, Mexico. These three coastal systems receive contaminants derived from local anthropogenic activities. Heat-shock proteins function to maintain protein integrity in the presence of stressors (such as heat or chemicals) and can be used as biomarkers of homeostatic alterations in polluted environments, whereas cytochrome P450 family members participate in steroid hormone synthesis and metabolism, and in xenobiotic transformation as a detoxification mechanism. The expression levels of both genes showed consistency in time and space, and presented a high overall correlation (r = 0.731, P < 0.001). Regardless of a high individual variability, both genes presented higher expression levels in the Urias Estuary (P < 0.001 and P < 0.05 for CYP1A and Hsp70, respectively), which was considered the most polluted among the three systems, especially during the rainy season (summer to fall). Gene expression levels were significantly associated with non-halogenated hydrocarbon concentrations in sediments during the sampling period (r = 0.686, P = 0.019 for CYP1A and r = 0.91, P < 0.001 for Hsp70), suggesting that both genes respond to chemicals in the environment. The results indicate that Mugil curema is a good candidate species to implement biomonitoring programs in tropical coastal environments.
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Follicular apoptosis in the mussel (Mytella strigata) as potential indicator of environmental stress in coastal ecosystems.
J Environ Sci Health A Tox Hazard Subst Environ Eng
PUBLISHED: 04-15-2010
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Follicular apoptosis in the tropical mussel Mytella strigata was assessed in three coastal lagoons located in the southern Gulf of California, Mexico. Mussels were collected from three coastal lagoons associated with different scenarios of anthropogenic stress during one year. The gonad of each mussel was dissected, weighed, and sampled for histology and apoptosis analysis by TUNEL labeling. Two apoptotic indices were used: the apoptotic index of cells (AIC) based on the number of follicular cells in apoptosis in one thousand cells counted per gonad, and the apoptotic index of follicles (AIF) based on the number of follicular cells per follicle per gonad. Both indices showed high association with each other for all developmental stages, although AIF seemed to better discriminate among sites. Higher AIF and AIC were observed at the Urias Estuary (1.6 and 1.5 respectively) ranked as highly polluted, followed by Ensenada del Pabellon (0.82 and 0.95 respectively), ranked as moderately polluted, and the Teacapan Estuary (0.57 and 0.76 respectively) ranked as slightly polluted. Our data indicate that the apoptotic index in tropical mussels could be a useful indicator of environmental stress in coastal ecosystems; however, the ecological relevance of follicular apoptosis in polluted environments needs further investigation.
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Effects of nursery preconditioning through mycorrhizal inoculation and drought in Arbutus unedo L. plants.
Mycorrhiza
PUBLISHED: 03-24-2010
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The influence of a water deficit treatment and mycorrhizal inoculation with Pisolithus tinctorius (Pers.) Coker and Couch on the water relations, gas exchange, and plant growth in Arbutus unedo L. plants was studied in order to evaluate the hardening process during the nursery period. The ability to withstand the adverse conditions after transplantation was also studied. Mycorrhizal and non-mycorrhizal seedlings of A. unedo were pot-grown for 4 months in a greenhouse (nursery period), during which time two irrigation treatments, well watered (100% water holding capacity, leaching 20% of the applied water) and deficit irrigation (50% of the well watered), were applied. Subsequently, the plants were transplanted to the field and well irrigated (transplanting period), after which and until the end of the experiment they received no water (establishment period). At the end of the nursery period, both water deficit and mycorrhizae were seen to have altered the plant morphology. Mycorrhizal plants had lower leaf area and improved leaf color parameters, while the water deficit increased root dry weight and the root/shoot ratio. Mycorrhizal plants had higher leaf water potential values than non-inoculated plants. Mycorrhizae increased stomatal conductance and photosynthesis values, especially in stressed plants. Drought led to an osmotic adjustment and a decrease in the leaf water potential values at turgor loss point in the mycorrhizal plants. Cell wall rigidity, measured as increased bulk modulus of elasticity, was decreased by the mycorrhizae effect. After transplanting, no differences were found in the water relations or gas exchange values between treatments. During the establishment period, the plants that had been exposed to both drought and mycorrhizae showed a better water status (higher leaf water and turgor potential values) and higher gas exchange values. In conclusion, water deficit and mycorrhizal inoculation of A. unedo plants in nursery produced changes in tissue water relations, gas exchange, and growth, related with the acclimation process in the seedlings, which could provide better resistance to drought and stress conditions following planting.
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Visualization of inositol 1,4,5-trisphosphate receptors on the nuclear envelope outer membrane by freeze-drying and rotary shadowing for electron microscopy.
J. Struct. Biol.
PUBLISHED: 03-21-2010
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The receptors for the second messenger InsP(3) comprise a family of closely related ion channels that release Ca(2+) from intracellular stores, most prominently the endoplasmic reticulum and its extension into the nuclear envelope. The precise sub-cellular localization of InsP(3)Rs and the spatial relationships among them are important for the initiation, spatial and temporal properties and propagation of local and global Ca(2+) signals, but the spatial organization of InsP(3)Rs in Ca(2+) stores is poorly characterized. Using nuclei isolated from insect Sf9 cells and freeze-dry rotary shadowing, we have addressed this by directly visualizing the cytoplasmic domain of InsP(3)R located on the cytoplasmic side of the nuclear envelope. Identification of approximately 15 nm structures as the cytoplasmic domain of InsP(3)R was indirectly supported by a marked increase in their frequency after transient transfections with cDNAs for rat types 1 and 3 InsP(3)R, and directly confirmed by gold labeling either with heparin or a specific anti-InsP(3)R antibody. Over-expression of InsP(3)R did not result in the formation of arrays or clusters with channels touching each other. Gold-labeling suggests that the channel amino terminus resides near the center of the cytoplasmic tetrameric quaternary structure. The combination of nuclear isolation with freeze-drying and rotary shadow techniques allows direct visualization of InsP(3)Rs in native nuclear envelopes and can be used to determine their spatial distribution and density.
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Hox gene expression in the embryonic genital system of the sea turtle Lepidochelys olivacea (Eschscholt, 1829), a species with temperature-dependent sex determination.
Gene Expr. Patterns
PUBLISHED: 03-11-2010
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Hox genes are conserved transcription factors which regulate embryonic morphogenesis and differentiation. For the first time, we examined the quantitative and spatial expression of two Hox 5 genes, HoxD11 and HoxA13, in the developing genital system of the olive ridley Lepidochelys olivacea, a species with temperature-dependent sex determination. Quantitative and spatial expression patterns of both genes suggest a role in the female pathway rather than the male pathway. For instance, both genes, especially HoxA13, were expressed in the undifferentiated gonad during the thermosensitive period at a female promoting temperature, and downregulated in the differentiated gonad. By contrast, expression of both genes was low in gonads incubated at a male promoting temperature and did not change significantly in the differentiated gonad. Furthermore, we found high expression levels of HoxA13 in the paramesonephric duct at the male promoting temperature but not at the female promoting temperature, suggesting a role for this Hox gene in the partial regression of the Müllerian duct in males.
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Early transcriptomic events in microdissected Arabidopsis nematode-induced giant cells.
Plant J.
PUBLISHED: 12-09-2009
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Root-knot nematodes differentiate highly specialized feeding cells in roots (giant cells, GCs), through poorly characterized mechanisms that include extensive transcriptional changes. While global transcriptome analyses have used galls, which are complex root structures that include GCs and surrounding tissues, no global gene expression changes specific to GCs have been described. We report on the differential transcriptome of GCs versus root vascular cells, induced in Arabidopsis by Meloidogyne javanica at a very early stage of their development, 3 days after infection (d.p.i.). Laser microdissection was used to capture GCs and root vascular cells for microarray analysis, which was validated through qPCR and by a promoter-GUS fusion study. Results show that by 3 d.p.i., GCs exhibit major gene repression. Although some genes showed similar regulation in both galls and GCs, the majority had different expression patterns, confirming the molecular distinctiveness of the GCs within the gall. Most of the differentially regulated genes in GCs have no previously assigned function. Comparisons with other transcriptome analyses revealed similarities between GCs and cell suspensions differentiating into xylem cells. This suggests a molecular link between GCs and developing vascular cells, which represent putative GC stem cells. Gene expression in GCs at 3 d.p.i. was also found to be similar to crown galls induced by Agrobacterium tumefaciens, a specialized root biotroph.
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Hexachlorobenzene induces cell proliferation and IGF-I signaling pathway in an estrogen receptor alpha-dependent manner in MCF-7 breast cancer cell line.
Toxicol. Lett.
PUBLISHED: 07-07-2009
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Hexachlorobenzene (HCB) is an organochlorine pesticide widely distributed in the biosphere. ERalpha regulates the expression of genes involved in growth and development, and plays an important role in breast cancer. The present study focuses attention on the effect of HCB (0.005, 0.05, 0.5, 5 microM) on cell proliferation in estrogen receptor alpha (ERalpha)(+) MCF-7, and ERalpha(-) MDA-MB-231 breast cancer cell lines. We also studied the insulin growth factor-I (IGF-I) signaling pathway in MCF-7 cells. HCB (0.005 and 0.05 microM) stimulated cell proliferation in MCF-7, but not in MDA-MB-231 cells. The pesticide increased apoptosis in MCF-7, at HCB (0.5 and 5 microM). At these doses, HCB induced the expression of the aryl hydrocarbon receptor (AhR)-regulated gene cytochrome P4501A1. MCF-7 cells exposed to HCB (0.005 and 0.05 microM) overexpressed IGF-IR and insulin receptor (IR). IRS-1-phosphotyrosine content was increased in a dose dependent manner. ICI 182,780 prevented HCB-induced cell proliferation and IGF-I signaling in MCF-7 cells incubated in phenol-red free media. In addition, HCB (0.005 microM) increased c-Src activation, Tyr537-ERalpha phosphorylation and ERalpha down-regulation. Taken together, our data indicate that HCB stimulation of cell proliferation and IGF-I signaling is ERalpha dependent in MCF-7 cells.
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Tensaw virus genome sequence and its relation to other Bunyaviridae.
Virus Genes
PUBLISHED: 06-07-2009
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Tensaw virus (TSV) belongs to the genus Orthobunyavirus within the Bunyaviridae family. Although TSV does not cause hemorrhagic fever as some other members of its family, serological studies have shown that serum from Florida residents react against TSV indicating viral infection in humans. In this study, the three RNA genome segments of a TSV isolated from Anopheles crucians mosquitoes collected in North Central Florida in 2006 and a TSV isolate obtained from the CDC, Fort Collins, were sequenced and compared to other Bunyaviridae. The placement of the TSVs within the Bunyamwera serogroup was confirmed by phylogenetic analysis of the inferred amino acid (aa) sequence of proteins coded by each of the RNA segments separately as well as by the combined tree of the same three inferred proteins. The N terminal glycoprotein (Gn) encoded by the M segment contained the 18 conserved Cysteines present in Bunyamwera and California serogroups, the two glycosylation sites, and residues considered potential proteolytic cleavage sites conserved in other Bunyaviridae. The TSV L protein displayed all the strictly conserved amino acids in the four conserved regions known to be catalytically active for the RNA dependent RNA polymerase transcriptase and replicase activities. The amino acid conservation between the two TSV viral isolates was 100, 99.4, and 99.6% for the S, M, and L segments, respectively.
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Analysis of the association of preeclampsia with polymorphisms of the INS, INSR and IRS1 genes in Mexican women.
Gynecol. Obstet. Invest.
PUBLISHED: 04-25-2009
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It has been proposed that preeclampsia is a metabolic syndrome of pregnancy. The polymorphisms PstI and MaeIII of INS, NsiI of INSR and Ala513Pro and Gly972Arg of IRS1 have been associated with metabolic syndrome; moreover, the products of these genes are functionally contiguous during insulin signaling. The aim of this study was to assess whether these polymorphisms are associated with preeclampsia.
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NADPH oxidase and hydrogen peroxide mediate insulin-induced calcium increase in skeletal muscle cells.
J. Biol. Chem.
PUBLISHED: 03-24-2009
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Skeletal muscle is one of the main physiological targets of insulin, a hormone that triggers a complex signaling cascade and that enhances the production of reactive oxygen species (ROS) in different cell types. ROS, currently considered second messengers, produce redox modifications in proteins such as ion channels that induce changes in their functional properties. In myotubes, insulin also enhances calcium release from intracellular stores. In this work, we studied in myotubes whether insulin stimulated ROS production and investigated the mechanisms underlying the insulin-dependent calcium increase: in particular, whether the late phase of the Ca2+ increase induced by insulin required ROS. We found that insulin stimulated ROS production, as detected with the probe 2,7-dichlorofluorescein diacetate (CM-H2DCFDA). We used the translocation of p47phox from the cytoplasm to the plasma membrane as a marker of the activation of NADPH oxidase. Insulin-stimulated ROS generation was suppressed by the NADPH oxidase inhibitor apocynin and by small interfering RNA against p47phox, a regulatory NADPH oxidase subunit. Additionally, both protein kinase C and phosphatidylinositol 3-kinase are presumably involved in insulin-induced ROS generation because bisindolylmaleimide, a nonspecific protein kinase C inhibitor, and LY290042, an inhibitor of phosphatidylinositol 3-kinase, inhibited this increase. Bisindolylmaleimide, LY290042, apocynin, small interfering RNA against p47phox, and two drugs that interfere with inositol 1,4,5-trisphosphate-mediated Ca2+ release, xestospongin C and U73122, inhibited the intracellular Ca2+ increase produced by insulin. These combined results strongly suggest that insulin induces ROS generation trough NADPH activation and that this ROS increase is required for the intracellular Ca2+ rise mediated by inositol 1,4,5-trisphosphate receptors.
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Analysis of transcripts from predicted open reading frames of Musca domestica salivary gland hypertrophy virus.
J. Gen. Virol.
PUBLISHED: 03-04-2009
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The Musca domestica salivary gland hypertrophy virus (MdSGHV) is a large dsDNA virus that infects and sterilizes adult houseflies. The transcriptome of this newly described virus was analysed by rapid amplification of cDNA 3-ends (3-RACE) and RT-PCR. Direct sequencing of 3-RACE products revealed 78 poly(A) transcripts containing 95 of the 108 putative ORFs. An additional six ORFs not amplified by 3-RACE were detected by RT-PCR. Only seven of the 108 putative ORFs were not amplified by either 3-RACE or RT-PCR. A series of 5-RACE reactions were conducted on selected ORFs that were identified by 3-RACE to be transcribed in tandem (tandem transcripts). In the majority of cases, the downstream ORFs were detected as single transcripts as well as components of the tandem transcripts, whereas the upstream ORFs were found only in tandem transcripts. The only exception was the upstream ORF MdSGHV084, which was differentially transcribed as a single transcript at 1 and 2 days post-infection (days p.i.) and as a tandem transcript (MdSGHV084/085) at 2 days p.i. Transcriptome analysis of MdSGHV detected splicing in the 3 untranslated region (3-UTR) and extensive heterogeneity in the polyadenylation signals and cleavage sites. In addition, 23 overlapping antisense transcripts were found. In conclusion, sequencing the 3-RACE products without cloning served as an alternative approach to detect both 3-UTRs and transcript variants of this large DNA virus.
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Expression from baculovirus and serological reactivity of the nucleocapsid protein of dolphin morbillivirus.
Vet. Microbiol.
PUBLISHED: 03-02-2009
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The nucleocapsid (N) protein of dolphin morbillivirus (DMV) was expressed from a baculovirus (Autographa californica nuclear polyhedrosis virus) vector and shown by SDS-PAGE and Western blot analysis to be about 57 kDa. Transmission electron microscopy revealed fully assembled nucleocapsid-like particles (NLPs) exhibiting the typical helical herringbone morphology. These NLPs were approximately 20-22 nm in diameter and varied in length from 50 to 100 nm. Purified DMV-N protein was used as antigen in an indirect ELISA (iELISA) and shown to react with rabbit and human antisera to measles virus (MV) and dog sera with antibodies to canine distemper virus (CDV). The iELISA was used for the demonstration of morbillivirus antibodies in the serum of cetaceans and manatees, showing potential as a serological tool for the mass screening of morbillivirus antibodies in marine mammals.
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Two viruses that cause salivary gland hypertrophy in Glossina pallidipes and Musca domestica are related and form a distinct phylogenetic clade.
J. Gen. Virol.
PUBLISHED: 01-15-2009
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Glossina pallidipes and Musca domestica salivary gland hypertrophy viruses (GpSGHV and MdSGHV) replicate in the nucleus of salivary gland cells causing distinct tissue hypertrophy and reduction of host fertility. They share general characteristics with the non-occluded insect nudiviruses, such as being insect-pathogenic, having enveloped, rod-shaped virions, and large circular double-stranded DNA genomes. MdSGHV measures 65x550 nm and contains a 124 279 bp genome (approximately 44 mol% G+C content) that codes for 108 putative open reading frames (ORFs). GpSGHV, measuring 50x1000 nm, contains a 190 032 bp genome (28 mol% G+C content) with 160 putative ORFs. Comparative genomic analysis demonstrates that 37 MdSGHV ORFs have homology to 42 GpSGHV ORFs, as some MdSGHV ORFs have homology to two different GpSGHV ORFs. Nine genes with known functions (dnapol, ts, pif-1, pif-2, pif-3, mmp, p74, odv-e66 and helicase-2), a homologue of the conserved baculovirus gene Ac81 and at least 13 virion proteins are present in both SGHVs. The amino acid identity ranged from 19 to 39 % among ORFs. An (A/T/G)TAAG motif, similar to the baculovirus late promoter motif, was enriched 100 bp upstream of the ORF transcription initiation sites of both viruses. Six and seven putative microRNA sequences were found in MdSGHV and GpSGHV genomes, respectively. There was genome. Collinearity between the two SGHVs, but not between the SGHVs and the nudiviruses. Phylogenetic analysis of conserved genes clustered both SGHVs in a single clade separated from the nudiviruses and baculoviruses. Although MdSGHV and GpSGHV are different viruses, their pathology, host range and genome composition indicate that they are related.
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Impaired synthesis and secretion of SopA in Salmonella Typhimurium dam mutants.
FEMS Microbiol. Lett.
PUBLISHED: 01-13-2009
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DNA adenine methylation regulates virulence gene expression in certain bacteria, including Salmonella Typhimurium. The aim of this study was to investigate the involvement of DNA adenine methylase (Dam) methylation in the expression and secretion of the SPI-1 effector protein SopA. For this purpose, SopA-FLAG-tagged wild-type and dam strains of Salmonella Typhimurium were constructed. The expression and secretion of SopA were determined in bacterial culture and in intracellular bacteria recovered from infected HEp-2 epithelial cells. Bacterial culture supernatants and pellets were used to investigate secreted proteins and cell-associated proteins, respectively. Western blot and quantitative reverse transcriptase PCR analysis showed that the dam mutant expresses lower levels of SopA than the wild-type strain. Interestingly, the strain lacking Dam synthesizes SopA under nonpermissive conditions (28 degrees C). In addition, SopA secretion was drastically impaired in the dam mutant. In vivo experiments showed that the intracellular Salmonella dam mutant synthesizes SopA although in lower amounts than the wild-type strain. Taken together, our results suggest that Dam methylation modulates the expression and secretion of SopA in Salmonella Typhimurium.
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Photocatalytic printing of inorganic nanopatterns via poly(styrene-block-carbosilane) copolymer thin films on titania substrates.
Chem. Commun. (Camb.)
PUBLISHED: 01-06-2009
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Well-defined, ordered arrays of nanoscale depressions were obtained in linear-brush-type polystyrene-block-polycarbosilane (PS-b-PCS) diblock copolymer thin films by acetone vapor annealing and silica nanodot arrays were directly obtained from such thin films deposited on a titania substrate by one-step exposure to UV light as a result of transformation of the PCS units to silica, driven by the photocatalytic activity of titania concurrent with removal of the organic matrix.
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Potassium sorbate controlled release from corn starch films.
Mater Sci Eng C Mater Biol Appl
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Active starch films with glycerol and potassium sorbate were obtained by casting. Native and acetylated corn starches, as well as the mixture of them in equal proportions were used and filmogenic suspensions with pH 4.5 were also prepared. Sorbate concentration decreased during film storage due to its oxidative degradation. Active films resulted more yellow and less transparent than films without sorbate. The minimum inhibitory concentration of sorbate resulted 0.3%, regardless of the starch type and the formulation pH. The use of antimicrobial package was more effective to prevent microbial growth on food surfaces than the use of conventional methods. Additive kinetic release was neither affected by the starch type nor by the formulation pH. Sorbate diffusion process was mathematically modeled satisfactorily. Active films were able to inhibit Candida spp., Penicillium spp., S. aureus and Salmonella spp. growth. Active films extended 21% the shelf life of refrigerated cheese, regardless of the formulation pH.
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Cleaved/associated TLR3 represents the primary form of the signaling receptor.
J. Immunol.
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TLR3 belongs to the family of intracellular TLRs that recognize nucleic acids. Endolysosomal localization and cleavage of intracellular TLRs play pivotal roles in signaling and represent fail-safe mechanisms to prevent self-nucleic acid recognition. Indeed, cleavage by cathepsins is required for native TLR3 to signal in response to dsRNA. Using novel Abs generated against TLR3, we show that the conserved loop exposed in LRR12 is the single cleavage site that lies between the two dsRNA binding sites required for TLR3 dimerization and signaling. Accordingly, we found that the cleavage does not dissociate the C- and N-terminal fragments, but it generates a very stable "cleaved/associated" TLR3 present in endolysosomes that recognizes dsRNA and signals. Moreover, comparison of wild-type, noncleavable, and C-terminal-only mutants of TLR3 demonstrates that efficient signaling requires cleavage of the LRR12 loop but not dissociation of the fragments. Thus, the proteolytic cleavage of TLR3 appears to fulfill function(s) other than separating the two fragments to generate a functional receptor.
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Is nicotine protective against Parkinsons disease? An experimental analysis.
CNS Neurol Disord Drug Targets
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Parkinsons disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta (SNc) and its projections. Reports show a lower incidence of PD in smokers compared to nonsmokers. Nicotine reduce motor symptoms of patients already diagnosed with PD. However, the mechanisms underlying the effects of nicotine in the dopamine (DA) depleted striatum remain elusive. This study evaluates the effects of chronic nicotine administration on PD motor symptoms in an attempt to mimic the chronic self-administration of nicotine in smokers. To achieve this, we used the 6-OHDA hemiparkinson rat model evaluating the amphetamine/apomorphine induced circling behavior, in rats whose daily water intake included nicotine. We found that chronic nicotine reduced amphetamine (AMPH) induced circling behavior by 40%, whereas apomorphine (APO) increased this behavior by 230%. High-performance liquid chromatography (HPLC) revealed that AMPH produced a 50% decrease of DA release in the intact hemisphere, while on the striatum of the lesioned side, receptor binding assays showed an increased affinity to D1 receptors and a concurrent decrease in D2 receptors. c-Fos activity showed through double labeling, that cell types involved in nicotine action were low threshold (LTS) and fast spiking (FS) inter-neurons, which increased in the DA-depleted striatum. We also observed an increase in the activity of D1 medium spiny neurons (D1 MSN), a striatal population with a major role in motor control. Our results show that chronic nicotine does not specifically protect against degeneration, but rather modifies DA receptor dynamics, suggesting that it could be used as a therapeutic element in PD pathology.
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Assessment of ?-H2AX levels in circulating tumor cells from patients receiving chemotherapy.
Front Oncol
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Circulating tumor cells (CTCs) are prognostic markers in a variety of solid tumor malignancies. The potential of CTCs to be used as a "liquid biopsy" to monitor a patients condition and predict drug response and resistance is currently under investigation. Using a negative depletion, enrichment methodology, CTCs isolated from the peripheral blood of breast cancer patients with stage IV breast cancer undergoing DNA damaging therapy with platinum-based therapy were enriched. The enriched cell suspensions were stained with an optimized labeling protocol targeting: nuclei, cytokeratins 8, 18, and 19, the surface marker CD45, and the presence of the protein ?-H2AX. As a direct or indirect result of platinum therapy, double-strand break of DNA initiates phosphorylation of the histone H2AX, at serine 139; this phosphorylated form is referred to as ?-H2AX. In addition to ?-H2AX staining in specific locations with the cell nuclei, consistent with previous reports and referred to as foci, more general staining in the cell cytoplasm was also observed in some cells suggesting the potential of cell apoptosis. Our study underscores the utility and the complexity of investigating CTCs as predictive markers of response to various therapies. Additional studies are ongoing to evaluate the diverse ?-H2AX staining patterns we report here which needs to be further correlated with patient outcomes.
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Serum levels of glutathione peroxidase 3 in overweight and obese subjects from central Mexico.
Arch. Med. Res.
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Overweight and obesity are considered complex entities in which there are alterations in the concentration of antioxidant enzymes. It has been reported that glutathione peroxidase 3 (GPx3), an extracellular enzyme involved in the reduction of both hydro- and lipoperoxides, shows changes both in gene expression and protein concentration in animal models for type 2 diabetes (T2D) and obesity, but the variability of GPx3 levels in different human populations and under different health conditions are currently unclear. We undertook this study to determine the GPx3 levels in overweight and obese subjects from central Mexico.
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Affibody-DyLight conjugates for in vivo assessment of HER2 expression by near-infrared optical imaging.
PLoS ONE
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Amplification of the HER2/neu gene and/or overexpression of the corresponding protein have been identified in approximately 20% of invasive breast carcinomas. Assessment of HER2 expression in vivo would advance development of new HER2-targeted therapeutic agents and, potentially, facilitate choice of the proper treatment strategy offered to the individual patient. We present novel HER2-specific probes for in vivo evaluation of the receptor status by near-infrared (NIR) optical imaging.
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Genome-wide analysis reveals that Smad3 and JMJD3 HDM co-activate the neural developmental program.
Development
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Neural development requires crosstalk between signaling pathways and chromatin. In this study, we demonstrate that neurogenesis is promoted by an interplay between the TGF? pathway and the H3K27me3 histone demethylase (HDM) JMJD3. Genome-wide analysis showed that JMJD3 is targeted to gene promoters by Smad3 in neural stem cells (NSCs) and is essential to activate TGF?-responsive genes. In vivo experiments in chick spinal cord revealed that the generation of neurons promoted by Smad3 is dependent on JMJD3 HDM activity. Overall, these findings indicate that JMJD3 function is required for the TGF? developmental program to proceed.
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Detection and characterization of bacterial symbionts in the Heteropteran, Blissus insularis.
FEMS Microbiol. Ecol.
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Dense populations of extracellular bacteria were detected in midgut crypts of the southern chinch bug, Blissus insularis Barber (Hemiptera: Blissidae). Examination by epifluorescent and transmission electron microscopy revealed that the bacteria covered the luminal surface of the crypts and filled the entire lumen. Attempts to culture the extracellular endosymbionts in various media failed. Sequencing and phylogenetic analyses of 16S rRNA gene clones obtained from insects of five Florida populations showed high nucleotide homology to either betaproteobacterial Burkholderia spp. (243 clones from five populations) or gammaproteobacterial Pseudomonas spp. (58 clones from one population). Using Burkholderia-specific primers, bacteria were detected in the egg, nymph, and adult stages. Fluorescent in situ hybridization with genus-specific oligonucleotide probes confirmed the localization of Burkholderia in the crypts. Quantitative real-time PCR showed that antibiotic treatments of nymphs significantly reduced the amount of Burkholderia 16S rRNA gene copies in chinch bugs sampled 11 days after the treatment. Furthermore, these treatments resulted in retarded development and high mortality of B. insularis, indicating a beneficial impact of Burkholderia on its host.
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Screening of R122H and N29I mutations in the PRSS1 gene and N34S mutation in the SPINK1 gene in Mexican pediatric patients with acute and recurrent pancreatitis.
Pancreas
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The studys objective was to assess the association between the PRSS1 R122H and N29I and the SPINK1 N34S mutations and acute pancreatitis (AP) and recurrent pancreatitis in Mexican pediatric patients.
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Cleavage of Toll-like receptor 3 by cathepsins B and H is essential for signaling.
Proc. Natl. Acad. Sci. U.S.A.
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Toll-like receptor (TLR) 3 is an endosomal TLR that mediates immune responses against viral infections upon activation by its ligand double-stranded RNA, a replication intermediate of most viruses. TLR3 is expressed widely in the body and activates both the innate and adaptive immune systems. However, little is known about how TLR3 intracellular trafficking and maturation are regulated. Here we show that newly synthesized endogenous TLR3 is transported through the ER and Golgi apparatus to endosomes, where it is rapidly cleaved. TLR3 protein expression is up-regulated by its own ligand, leading to the accumulation of its cleaved form. In agreement with its proposed role as a transporter, UNC93B1 expression is required for TLR3 cleavage and signaling. Furthermore, TLR3 signaling and cleavage are sensitive to cathepsin inhibition. Cleavage occurs between aa 252 and 346, and results in a functional receptor that signals upon activation. A truncated form of TLR3 lacking the N-terminal 345 aa also signals from acidic compartments in response to ligand activation. Screening of the human cathepsin family by RNA interference identified cathepsins B and H as key mediators of TLR3 processing. Taken together, our data indicate that TLR3 proteolytic processing is essential for its function, and suggest a mechanism of tight control of TLR3 signaling and thus immunity.
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A new species of Euryhaliotrematoides Plaisance & Kritsky, 2004 (Monogenea: Dactylogyridae) from the gills of the spotted rose snapper Lutjanus guttatus (Steindachner) (Perciformes: Lutjanidae).
Syst. Parasitol.
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Euryhaliotrematoides mehen n. sp. (Monogenea: Dactylogyridae) is described based on specimens collected from the gills of the spotted rose snapper Lutjanus guttatus (Steindachner) off Mazatlan, Sinaloa, on the northwestern coast of Mexico. The synapomorphy of this genus is the presence of a funnel-shaped base of the male copulatory organ. This new species differs from all other species of the genus by possessing a male copulatory organ with a base with a thickened margin and a membranous accessory piece resembling a scarf and covering about 60% of its distal region. PCR products of the 28S rRNA (831 bp) and 18S rRNA (662 bp) genes were sequenced and submitted to GenBank (accession numbers HQ615997 and JF938069, respectively). BLASTn searches revealed no 100% identical hits with the previously registered monogenean sequences.
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Identification of a tubulin-? gene specifically expressed in testis and adductor muscle during stable reference gene selection in the hermaphrodite gonad of the lions paw scallop Nodipecten subnodosus.
Mar Genomics
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For non-model species, as many used for aquaculture, with minimal or no genomic information, relative quantification of gene expression studies requires preliminary research including the isolation of potential reference genes and the identification of those stably expressed under the biological conditions of interest. Here we report on the isolation of five partial gene sequences from gonad tissue cDNA in the functional hermaphrodite scallop Nodipecten subnodosus to be evaluated as reference genes: 18S-rRNA, riboprotein l8 (rp-l8), actin-? (act-?), elongation factor 1? (ef-1?) and alpha-tubulin-? (tub-?). We found that 18S-rRNA was stably expressed independently of the priming method used to reverse transcribe RNA to cDNA, oligo-dT or random hexamer. Stability analysis for the five putative reference genes with geNorm and NormFinder indicated that 18S together with rp-l8 were the most stable genes for normalization of gene expression during gonad development in both, male and female sexual regions of the hermaphrodite N. subnodosus. The least stable gene was tub-?, showing a biased expression profile between sexual regions of the gonad, therefore this gene was analyzed thereafter as a target gene together with vitellogenin (vit) and a DEAD-box RNA helicase (dbx) gene. Relative expression, estimated by normalization with the combination of 18S and rp-l8 as reference genes, indicated that as gonad development advanced two of the target genes were up-regulated, tub-? in the male region and vit in the female region. Whereas an increased expression was expected during development for vit for its known role in vitellogenesis, the increased expression of tub-? in the male sexual region was unexpected, and pointed toward this gene being a testis-specific ?-tubulin isotype. Further analyses of gene expression among tissues indicated that tub-? is specifically and highly expressed in the male gonad, although expression in adductor muscle was also observed at significantly lower levels. The existence of testis specific ?- and ?-tubulins has been previously reported in other taxa, relating their function to sperm axoneme formation. Tissue-specific tubulin genes, particularly their promoters, have recently found an application as native promoters for transgene tissue-specific expression in research and reproductive control of insect plagues. The third target gene, a putative member of the DEAD-box RNA helicase family (dbx), showed no changes in expression during gonad development or between sexual regions, therefore it was chosen to discuss the different statistical inferences resulting from the arbitrary use of randomly chosen reference genes when normalizing gene expression.
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