ABSTRACT
La elección de este caso se debe a la complejidad diagnóstica y terapéutica que ha supuesto y la escasa evolución positiva pese a los múltiples recursos humanos, profesionales, terapéuticos y farmacológicos invertidos. Se trata de un paciente con alteraciones de la conducta asociadas a un trastorno generalizado del desarrollo con sintomatología comórbida de TDAH, que posee un mundo imaginario que roza el pensamiento delirante. El paranoidismo, sin ninguna temporalidad en los hechos, le conduce a tener que vengarse en forma de pasos al acto violentos en el momento en que él cree ser la víctima. La evolución es tórpida, resaltando que esta mecánica de interpretación del mundo se presentaba generalmente como egosintónica, siendo muy escasas las ocasiones en que el paciente manifestaba, con llanto, un deseo de no querer hacer daño a nadie
We have chosen this case because of the therapeutic and diagnostic complexity. Furthermore the limited positive progress despite the numerous human, professional, therapeutic and pharmacological resources employed. It is about a patient who suffers behavioral disorders associated with a pervasive developmental disorder with comorbid symptomatology of ADHD, that owns an imaginary world which is close to delirious thought. The paranoid behavior with no temporality in the acts, drives him to revenge in a step way to the violent act in the moment he thinks himself as a victim. The evolution is torpid, standing out this mechanical world interpretation was usually presented as egosyntonic, being really limited the occasions in which the patient declared crying a wish of hurting no one
Subject(s)
Humans , Child , Child Development Disorders, Pervasive/diagnosis , Child Development Disorders, Pervasive/psychology , Child Behavior/psychology , Antisocial Personality Disorder/psychology , Comorbidity , Child Development Disorders, Pervasive/epidemiology , PsychopathologyABSTRACT
Heat shock protein A8 (HSPA8) is a highly conserved member of the Hsp70 family, which is expressed in oviductal cells, translocated into oviductal fluid, and becomes attached to the sperm surface during sperm transport. Previous research has shown that HSPA8 supports mammalian sperm viability during in vitro incubation at both 5 °C and body temperature. The present series of experiments was designed to explore the possibility that bovine recombinant HSPA8 might therefore protect bull spermatozoa during cryopreservation through its beneficial effects on the sperm plasma membrane. Soy-based cryopreservation media were used in these experiments. The effects of HSPA8 addition before freezing were examined at concentrations ranging from 0.2 to 6.4 µg/mL, whereas the effects of postthaw HSPA8 addition were tested between 0.2 and 12.8 µg/mL. When bull spermatozoa (from beef and dairy breeds) were frozen in the presence of HSPA8, beneficial but complex effects on postthaw viability were observed. Low HSPA8 concentrations (0.2 and 0.4 µg/mL) resulted in significantly reduced postthaw sperm viability, but concentrations above 0.8 µg/mL improved plasma membrane integrity. If HSPA8 was added to spermatozoa after thawing, outcomes were also biphasic and beneficial effects on viability were only seen if the HSPA8 concentration exceeded 3.2 µg/mL. Beneficial effects were significantly more apparent with beef rather than dairy breeds. When HSPA8 was used in combination with cholesterol-loaded cyclodextrin, spermatozoa from the beef breeds showed significantly lower apoptotic effects. This was not observed with the dairy breeds.
Subject(s)
Cell Membrane/ultrastructure , HSC70 Heat-Shock Proteins/pharmacology , Semen Preservation/veterinary , Spermatozoa/ultrastructure , Animals , Apoptosis/drug effects , Cattle , Cell Membrane/metabolism , Cryopreservation/methods , Cryopreservation/veterinary , Cyclodextrins/pharmacology , HSC70 Heat-Shock Proteins/metabolism , HSC70 Heat-Shock Proteins/physiology , Male , Recombinant Proteins/pharmacology , Semen Analysis/veterinary , Semen Preservation/methods , Spermatozoa/metabolism , Spermatozoa/physiologyABSTRACT
The aim of this study was to assess biologically safer components as alternatives to egg yolk for the frozen storage of ram semen using casein, coconut or palm oil in either Salamon's diluent (S) or a swim-up medium (SU). Ejaculates were frozen as pellets and sperm motility (subjectively) and acrosome integrity (FITC-PNA/PI) by flow cytometry were assessed at 0, 3 and 6h after thawing and incubation at 37°C. Three experiments were done: different concentrations of palm oil (5%, 10% and 20%); casein added as emulsifier and protective agent; and differences between egg yolk, coconut and palm oil in S and SU. 20% of oil added to SU accounted for a lesser percentage (P<0.05) of motile cells compared to rest while no differences were found between different oil levels on viable cells. When casein was added to diluents containing 5% of palm oil, no differences were found between palm or casein (P>0.05). No differences were found when S and SU were compared neither as groups nor between S alone and containing coconut or palm oil; however, SU alone yielded less motility than SU 5% coconut. However, in both groups, S and SU, egg yolk accounted for the greatest values in both bases. These results indicate that none of biologically safer media components (casein, palm or coconut oil) used in this study maintained the function of ram spermatozoa after freeze-thawing better than S-containing egg yolk. The application of vegetable oils as substitutes for egg yolk in diluents for the cryopreservation of ram spermatozoa requires further research.
Subject(s)
Caseins/pharmacology , Cryopreservation/methods , Plant Oils/pharmacology , Semen Preservation/methods , Sheep, Domestic , Spermatozoa/drug effects , Animals , Coconut Oil , Cryopreservation/veterinary , Cryoprotective Agents/pharmacology , Emulsifying Agents/pharmacology , Freezing/adverse effects , Male , Palm Oil , Semen Analysis/methods , Semen Analysis/veterinary , Semen Preservation/adverse effects , Semen Preservation/veterinary , Sheep, Domestic/physiology , Sperm Motility/drug effects , Sperm Motility/physiology , Spermatozoa/physiologyABSTRACT
Centrifugal countercurrent distribution (CCCD) in an aqueous two-phase system (TPS) is a resolute technique revealing sperm heterogeneity and for the estimation of the fertilizing potential of a given semen sample. However, separated sperm subpopulations have never been tested for their fertilizing ability yet. Here, we have compared sperm quality parameters and the fertilizing ability of sperm subpopulations separated by the CCCD process from ram semen samples maintained at 20°C or cooled down to 5°C. Total and progressive sperm motility was evaluated by computer-assisted analysis using a CASA system and membrane integrity was evaluated by flow cytometry by staining with CFDA/PI. The capacitation state, staining with chlortetracycline, and apoptosis-related markers, such as phosphatidylserine (PS) translocation detected with Annexin V, and DNA damage detected by the TUNEL assay, were determined by fluorescence microscopy. Additionally, the fertilizing ability of the fractionated subpopulations was comparative assessed by zona binding assay (ZBA). CCCD analysis revealed that the number of spermatozoa displaying membrane and DNA alterations was higher in samples chilled at 5°C than at 20°C, which can be reflected in the displacement to the left of the CCCD profiles. The spermatozoa located in the central and right chambers (more hydrophobic) presented higher values (P<0.01) of membrane integrity, lower PS translocation (P<0.05) and DNA damage (P<0.001) than those in the left part of the profile, where apoptotic markers were significantly increased and the proportion of viable non-capacitated sperm was reduced. We have developed a new protocol to recover spermatozoa from the CCCD fractions and we proved that these differences were related with the fertilizing ability determined by ZBA, because we found that the number of spermatozoa attached per oocyte was significantly higher for spermatozoa recovered from the central and right chambers, in both types of samples. This is the first time, to our knowledge that sperm recovered from a two-phase partition procedure are used for fertilization assays. These results open up new possibilities for using specific subpopulations of sperm for artificial insemination or in vitro fertilization, not only regarding better sperm quality but also certain characteristics such as subpopulations enriched in spermatozoa bearing X or Y chromosome that we have already isolated or any other feature.
Subject(s)
Chemical Fractionation/methods , Countercurrent Distribution/methods , Flow Cytometry/methods , Spermatozoa/chemistry , Spermatozoa/physiology , Analysis of Variance , Animals , Annexin A5/analysis , Apoptosis/physiology , Biomarkers , Centrifugation , DNA Damage , Male , Oocytes/metabolism , Phosphatidylserines/analysis , Reproductive Techniques, Assisted , Sheep, Domestic , Specimen Handling , Sperm Capacitation/physiology , Sperm Motility/physiology , Spermatozoa/metabolism , TemperatureABSTRACT
Egg yolk and milk are the 2 major membrane cryoprotectants commonly used in freezing media for the long-term preservation of semen (alone or in combination with others). However, in recent years, there have been increasing arguments against the use of egg yolk or milk because of the risk of introducing diseases through the use of cryopreserved semen. In this study, we analyzed the protective effect of lecithin as an alternative to egg yolk for the cryopreservation of ram semen, using a range of functional markers for sperm viability, motility, apoptosis, and mitochondrial functionality analyses (mitochondrial inner membrane surface [MIMS], mitochondrial inner membrane potential [MIMP], and cell membrane potential) as methods of assessment in samples diluted in 3 different media: Tris-citrate-glucose as control and 2 media supplemented with soy lecithin or egg yolk. The results showed that lecithin was able to effectively protect certain sperm quality characteristics against freezing-induced damage. However, lecithin induced loss of mitochondrial membrane potential or mitochondrial loss that was not reflected by modifications in sperm motility in fresh semen. MIMS and MIMP values decreased in thawed lecithin-treated samples, concomitant with a lower (P < .05) percentage of total and progressively motile cells, compared with those in egg yolk-containing samples. Further incubation of thawed samples revealed changes in motility and mitochondrial functionality that otherwise would not have been detected. These results indicated that lecithin may have affected the inner mitochondrial membrane in frozenthawed spermatozoa and confirmed that sublethal damages that seriously affect sperm functionality, not detected by classic sperm quality analyses, can be evidenced by changes in the inner mitochondrial membrane surface. These findings strengthen the relationship between mitochondrial membrane potential and motility and show that the mitochondrial alterations induced by the cryopreservation process could be specific targets for the improvement of semen cryopreservation protocols.
Subject(s)
Lecithins/adverse effects , Membrane Potential, Mitochondrial/drug effects , Semen Preservation/methods , Spermatozoa/drug effects , Animals , Cryopreservation/methods , Cryoprotective Agents , Egg Yolk , Freezing , Male , Mitochondria/drug effects , Mitochondria/physiology , Sheep, Domestic , Glycine max/chemistry , Sperm Motility/drug effectsABSTRACT
Two monoclonal antibodies (Nilo1 and Nilo2) were generated after immunization of hamsters with E13.5 olfactory bulb-derived mouse neurospheres. They are highly specific for neural stem and early progenitor cell surface antigens. Nilo positive cells present in the adult mouse subventricular zone (SVZ) were able to initiate primary neural stem cell cultures. Moreover, these antibodies added to neurosphere cultures induced proliferation arrest and interfered with their differentiation. In the lateral ventricles of adult mice, Nilo1 stained a cell subpopulation lining the ventricle and cells located in the SVZ, whereas Nilo2 stained a small population associated with the anterior horn of the SVZ at the beginning of the rostral migratory stream. Co-staining of Nilo1 or Nilo2 and neural markers demonstrated that Nilo1 identifies an early neural precursor subpopulation, whereas Nilo2 detects more differentiated neural progenitors. Thus, these antibodies identify distinct neurogenic populations within the SVZ of the lateral ventricle.
Subject(s)
Antibodies, Monoclonal/pharmacology , Neural Stem Cells/cytology , Stem Cell Niche/cytology , Animals , Antibodies, Monoclonal/immunology , Antibody Specificity , Antigens, Surface/immunology , Antigens, Surface/metabolism , Biomarkers/metabolism , Cell Differentiation , Cell Proliferation , Cells, Cultured , Cerebral Ventricles/cytology , Cricetinae , Embryo, Mammalian , Immunohistochemistry , Mice , Neural Stem Cells/metabolism , Olfactory Bulb/cytology , Stem Cell Niche/metabolismABSTRACT
The objectives of this study were two. First, to compare three base media with different sugar composition as an initial step to achieve a good chemically-defined extender for ram sperm refrigeration. The second one, to determine which sperm quality parameters may be more useful for revealing differences between sperm samples. One medium contained 200 mM sucrose and 2.8 mM glucose (SM), another only disaccharides (D) such as sucrose, trehalose, maltose and lactose (75 mM each); and the third one (D+M) included a mix of monosaccharides (50 mM glucose, 20 mM fructose and 20 mM galactose,) and the same disaccharides as in D (50 mM each). Ram semen samples diluted in the mentioned media were refrigerated at 5°C for 1 h, and rewarmed upto 37°C in order to mimic the temperature in the female reproductive tract. Addition of monosaccharides to the extender did not produce a better preservation of motility or viability after cooling. The supplementation with other disaccharides apart from sucrose did not enhance the viability either. Thus, after cooling and rewarming, there were no significant differences in sperm viability (membrane integrity evaluated by CFDA/PI staining) or the percentage of progressive motile and rapid sperm (evaluated by CASA) between the three media. However, the percentage of viable non-capacitated sperm evaluated by the chlortetracycline (CTC) assay was higher and sperm oxygen consumption was lower in SM than in D and in D+M. Although the apoptosis-like markers [phosphatidylserine exposure assessed by Annexin V/CFDA staining and DNA-damage evaluated by TUNEL assay] showed a continuous increment throughout the process with all diluents, the percentage of sperm with damaged DNA at the end of the process was significantly lower in SM than in the other two media (p < 0.01). On the basis of these results, we would make two recommendations: the use of an extender supplemented only with sucrose and glucose for ram sperm refrigeration; the inclusion of non-conventional methods such as oxygen consumption measure, evaluation of capacitation state and apoptosis-like markers for revealing differences between sperm samples.
Subject(s)
Cryopreservation/veterinary , Semen Preservation/veterinary , Sheep/physiology , Spermatozoa/drug effects , Spermatozoa/physiology , Animals , Cell Survival , Cold Temperature , DNA Fragmentation , In Situ Nick-End Labeling , Male , Oxygen Consumption , Phosphatidylserines/metabolism , Sperm Motility , Time FactorsABSTRACT
During embryo neurogenesis, neurons that originate from stem cells located in the forebrain subventricular zone (SVZ) continuously migrate to the olfactory bulb (OB). However, other authors describe the occurrence of resident stem cells in the OB. In the present work we report that the absence of tumor suppressor protein p53 increases the number of neurosphere-forming cells and the proliferation of stem cells derived from 13.5-day embryo OB. Interestingly, differentiation of p53 knockout-derived neurospheres was biased toward neuronal precursors, suggesting a role for p53 in the differentiation process. Moreover, we demonstrate the relevance of p53 in maintaining chromosomal stability in response to genotoxic insult. Finally, our data show that neurosphere stem cells are highly resistant to long-term epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) deprivation in a p53-independent fashion, and they preserve their differentiation potential. Thus, these data demonstrate that p53 controls the proliferation, chromosomal stability and differentiation pattern of embryonic mouse olfactory bulb stem cells.
Subject(s)
Cell Differentiation/genetics , Neurogenesis/genetics , Neurons/physiology , Stem Cells/physiology , Tumor Suppressor Protein p53/physiology , Animals , Annexins/metabolism , Cell Cycle/genetics , Cell Proliferation/drug effects , Cell Proliferation/radiation effects , Cells, Cultured , Chromosome Aberrations/radiation effects , Embryo, Mammalian , Epidermal Growth Factor/deficiency , Fibroblast Growth Factor 2/deficiency , Flow Cytometry/methods , Gene Expression Regulation, Developmental/drug effects , Gene Expression Regulation, Developmental/genetics , Glial Fibrillary Acidic Protein/metabolism , In Situ Nick-End Labeling/methods , Mice , Mice, Inbred C57BL , Mice, Knockout , Neurogenesis/drug effects , Neurons/drug effects , Neurons/radiation effects , O Antigens/metabolism , Olfactory Bulb/cytology , Protein Binding , Time Factors , Tubulin/metabolism , Tumor Suppressor Protein p53/deficiency , X-Rays/adverse effectsABSTRACT
No disponible
Subject(s)
Male , Adult , Humans , Hemochromatosis/genetics , Histocompatibility Antigens Class I/genetics , Membrane Proteins/geneticsSubject(s)
Ferritins/deficiency , Hemochromatosis/blood , Blood Donors , Female , Ferritins/blood , Hemochromatosis/diagnosis , Hemochromatosis/genetics , Hemochromatosis Protein , Histocompatibility Antigens Class I/genetics , Humans , Membrane Proteins/genetics , Metrorrhagia/complications , Middle AgedABSTRACT
INTRODUCTION: A total of 215 schyzophrenic patients according to DSM-IV criteria in treatment on risperidone were included in an open label postmarketing surveillance 18 months study to evaluate safety and effectivity of the drug in preventing relapses. METHODS: The Brief Psychiatric Rating Scale, Global Functional Assessment Scale and the Clinical Global Impression were used to assess. Safety was evaluated by the UKU subscale for neurological side effects. RESULTS: A 82.1% of the patients continued risperidone medication without relapse during the 18 month period. Risperidone was used at a mean dosage of 5.69 2.41 mg/d. DISCUSSION: Patients improved psychotic symptoms and global activity, and significant reductions were observed in mean total UKU subscale for neurological side effect score. 91.7% of the patients did not report any adverse event; only 2 (1.2%) patients dropt out because of intolerance.
Subject(s)
Antipsychotic Agents/therapeutic use , Psychotic Disorders/etiology , Psychotic Disorders/prevention & control , Risperidone/therapeutic use , Schizophrenia , Schizophrenic Psychology , Adolescent , Adult , Aged , Antipsychotic Agents/administration & dosage , Antipsychotic Agents/adverse effects , Consumer Product Safety , Female , Follow-Up Studies , Humans , Male , Middle Aged , Prospective Studies , Risperidone/administration & dosage , Risperidone/adverse effects , Secondary Prevention , Treatment OutcomeABSTRACT
Introducción: En un estudio abierto, observacional, de 18 meses de duración, 215 pacientes diagnosticados de esquizofrenia (DSM-IV), fueron tratados con risperidona con el objeto de evaluar la seguridad y efectividad del fármaco en la prevención de las recaídas. Material y métodos: La efectividad de la risperidona se evaluó mediante el tiempo transcurrido hasta la recaída e instrumentos de medida como la Escala Breve de Valoración Psiquiátrica, la Escala de Evaluación de la Actividad Global e Impresión Clínica Global. La seguridad se valoró mediante la subescala UKU para efectos adversos de tipo neurológico. Resultados: El 82,1 por ciento de los pacientes permanecieron en tratamiento con risperidona durante los 18 meses del estudio sin presentar recaídas. La dosis media de risperidona utilizada fue de 5,69 ñ 2,41 mg/d. Discusión: El tratamiento con risperidona mejoró significativamente la sintomatología psicótica y la actividad global de los pacientes; también se detectaron disminuciones estadísticamente significativas en la puntuación media total de la subescala UKU. El 91,7 por ciento de los pacientes no comunicó ningún efecto adverso; únicamente dos (1,2 por ciento) abandonaron el estudio de forma prematura por intolerancia (AU)
Subject(s)
Middle Aged , Adult , Adolescent , Aged , Male , Female , Humans , Schizophrenia , Schizophrenic Psychology , Antipsychotic Agents , Risperidone , Treatment Outcome , Psychotic Disorders , Recurrence , Prospective Studies , Consumer Product Safety , Follow-Up StudiesABSTRACT
One of the problems associated with the modern biomaterials used in prostheses is osteolysis, which, although its exact origin is unknown, has been associated with wear particles. Osteoblasts seem to participate directly in this phenomenon. This paper investigates in vitro cellular response to the wear particles from the metal substrate and ceramic covering (alpha-alumina) of a new titanium yttrium aluminum alloy, MA 956, that has been proposed as a biomaterial because of its exceptional mechanical and electrochemical properties. The effect of different sizes (10 and 80 microm) of MA 956 and alpha-alumina particles on osteoblast function was studied in primary human bone cell cultures. Cells were harvested from trabecular bone fragments obtained during knee arthroplasty. Osteoblastic cell response to the particles was measured by assaying C-terminal type I procollagen (PICP), alkaline phosphatase, and osteocalcin secretion, with and without 1.25(OH)(2)D(3) stimulation, in the cell-conditioned medium. Both sizes of MA 956 and alpha-alumina particles decreased PICP secretion in nonstimulated osteoblastic cells, but this secretion was not affected in the cultures stimulated with 1.25(OH)(2)D(3). Only the 10 microm alpha-alumina particles inhibited alkaline phosphatase activity in 1.25(OH)(2)D(3)-stimulated and nonstimulated cultures. The rise in osteocalcin levels after 1.25(OH)(2)D(3) stimulation was lower in the presence of the 10 microm MA 956 particles than in the presence of alpha-alumina particles. Although both materials seem to have directly affected in vitro osteoblastic cell function, the increase in osteocalcin levels after 1.25(OH)(2)D(3) stimulation was lower after exposure to MA 956 particles than the increase observed after exposure to alpha-alumina particles. Therefore, it does not seem that osteocalcin stimulated bone resorption, suggesting that MA 956 would be less likely to provoke osteolysis.