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1.
Neuroophthalmology ; 44(1): 45-48, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32076450

RESUMEN

Homonymous hemianopia is frequently associated with retrochiasmal lesions. Vascular etiology is the most common and usually evident on magnetic resonance imaging. When the results of neuroimaging are normal, there are other etiologies that we should consider, like nonketotic hyperglycemia (NKH). We report a 62-year-old female diabetic patient with headache, colour vision and sudden homonymous inferior quadrantanopia and elevated blood sugar levels with normal pH. The neuroimaging was normal and the visual lost improved after the correction of the hyperglycemia. NKH should be considered in patients with sudden and transient hemianopia and normal neuroimaging.

2.
Reprod Fertil Dev ; 31(4): 787-795, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30562475

RESUMEN

Metformin is a leading antidiabetic drug that is used worldwide in the treatment of diabetes mellitus. This biguanide exerts metabolic and pleiotropic effects in somatic cells, although its invitro actions on human spermatozoa remain unknown. The present study investigated the effects of metformin on human sperm function. Human spermatozoa were incubated in the presence or absence of 10mM metformin for 8 or 20h, and motility was measured by computer-aided sperm analysis (CASA); other parameters were evaluated by flow cytometry. Metformin significantly reduced the percentage of motile, progressive and rapid spermatozoa and significantly decreased sperm velocity. Metformin did not affect viability, mitochondrial membrane potential (MMP) or mitochondrial superoxide anion generation of human spermatozoa at any time studied. However, metformin clearly inhibited the protein kinase (PK) A pathway and protein tyrosine phosphorylation at 8 and 20h, key regulatory pathways for correct sperm function. In summary, metformin treatment of human spermatozoa had a detrimental effect on motility and inhibited essential sperm signalling pathways, namely PKA and protein tyrosine phosphorylation, without affecting physiological parameters (viability, MMP, mitochondrial superoxide anion generation). Given the growing clinical use of metformin in different pathologies in addition to diabetes, this study highlights an adverse effect of metformin on spermatozoa and its relevance in terms of human fertility in patients who potentially could be treated with metformin in the future.


Asunto(s)
Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Hipoglucemiantes/farmacología , Metformina/farmacología , Mitocondrias/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Motilidad Espermática/efectos de los fármacos , Humanos , Masculino , Mitocondrias/metabolismo , Fosforilación/efectos de los fármacos , Análisis de Semen , Espermatozoides/efectos de los fármacos , Espermatozoides/metabolismo , Superóxidos/metabolismo , Tirosina/metabolismo
3.
Reprod Fertil Dev ; 30(2): 297-306, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-28679463

RESUMEN

Equine cumulus-oocyte complexes (COCs) are classified as compact (cCOC) or expanded (eCOC) and vary in their meiotic competence. This difference could be related to divergent glucose metabolism. To test this hypothesis in the present study, eCOCs, cCOCs and expanded or compact mural granulosa cells (EC and CC respectively) were matured in vitro for 30h, at which time maturation rate, glucose metabolism and the expression of genes involved in glucose transport, glycolysis, apoptosis and meiotic competence were determined. There were significant differences between eCOCs and cCOCs in maturation rate (50% vs 21.7% (n=192 and 46) respectively; P<0.001), as well as mean (±s.e.m.) glucose consumption (1.8±0.5 vs 27.9±5.9 nmol per COC respectively) and pyruvate (0.09±0.01 vs 2.4±0.8 nmol per COC respectively) and lactate (4.7±1.3 vs 64.1±20.6 nmol per COC respectively; P<0.05 for all) production. Glucose consumption in EC and CC did not differ significantly. Expression of hyaluronan-binding protein (tumour necrosis factor alpha induced protein 6; TNFAIP6) was increased in eCOCs and EC, and solute carrier family 2 member 1 (SLC2A1) expression was increased in eCOCs, but there were no differences in the expression of glycolysis-related enzymes and solute carrier family 2 member 3 (SLC2A3) between the COC or mural granulosa cell types. The findings of the present study demonstrate that metabolic and genomic differences exist between eCOCs and cCOCs and mural granulosa cells in the horse.


Asunto(s)
Células del Cúmulo/metabolismo , Glucosa/metabolismo , Glucólisis , Caballos/metabolismo , Meiosis , Oocitos/metabolismo , Animales , Apoptosis , Células Cultivadas , Células del Cúmulo/patología , Femenino , Regulación del Desarrollo de la Expresión Génica , Glucólisis/genética , Técnicas de Maduración In Vitro de los Oocitos , Meiosis/genética , Metabolómica/métodos , Microscopía Fluorescente , Oocitos/patología , Espectroscopía de Protones por Resonancia Magnética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
4.
Reprod Domest Anim ; 53(1): 243-248, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29110393

RESUMEN

Current in vitro embryo production protocols in the Iberian red deer (Cervus elaphus hispanicus) need to be optimized; oocyte harvesting in situ followed by overnight holding could reduce the human effort and shipping costs. In our work, post-mortem ovaries were retrieved, and the oocytes were harvested and allocated to G1 group (good quality) or G2 + G3 group (low quality). The oocytes were separately subjected to immediate in vitro maturation (IVM) or held overnight in a holding medium composed of 40% of TCM 199 with Earle's salts, 40% TCM 199 with Hanks' salts and 20% fetal bovine serum (FBS), at room temperature (16 hr). In vitro maturation was carried out in a basal medium supplemented or not with 50 ng/ml of epidermal growth factor (EGF). Our data showed that addition of EGF to the maturation medium increases the percentage of G1 oocytes reaching metaphase II (3.9% vs. 50%, basal vs. EGF; p < .001) and decreased their degeneration rate (69.9% vs. 22.2%, basal vs. EGF; p < .01) when oocytes were immediately matured. Overnight holding increased the meiotic competence of G1 oocytes (37.5% matured in basal medium) and EGF increased prophase arrest in G2 + G3 oocytes (16.1% vs. 38.8% in germinal vesicle [GV] stage in basal medium vs. EGF added medium; p < .05). Our data demonstrate that oocyte holding can be used in Iberian red deer oocytes. Interestingly, EGF addition increases the oocytes' meiotic competence in immediately matured oocytes but not after oocyte holding depending upon initial oocyte quality.


Asunto(s)
Ciervos/fisiología , Factor de Crecimiento Epidérmico/farmacología , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Oocitos/efectos de los fármacos , Animales , Medios de Cultivo , Femenino , Oocitos/crecimiento & desarrollo
5.
Reproduction ; 149(1): 87-99, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25349439

RESUMEN

Repeatable methods for IVF have not been established in the horse, reflecting the failure of standard capacitating media to induce changes required for fertilization capacity in equine sperm. One important step in capacitation is membrane cholesterol efflux, which in other species is triggered by cholesterol oxidation and is typically enhanced using albumin as a sterol acceptor. We incubated equine sperm in the presence of calcium, BSA, and bicarbonate, alone or in combination. Bicarbonate induced an increase in reactive oxygen species (ROS) that was abolished by the addition of calcium or BSA. Bicarbonate induced protein tyrosine phosphorylation (PY), even in the presence of calcium or BSA. Incubation at high pH enhanced PY but did not increase ROS production. Notably, no combination of these factors was associated with significant cholesterol efflux, as assessed by fluorescent quantitative cholesterol assay and confirmed by filipin staining. By contrast, sperm treated with methyl-ß-cyclodextrin showed a significant reduction in cholesterol levels, but no significant increase in PY or ROS. Presence of BSA increased sperm binding to bovine zonae pellucidae in all three stallions. These results show that presence of serum albumin is not associated with a reduction in membrane cholesterol levels in equine sperm, highlighting the failure of equine sperm to exhibit core capacitation-related changes in a standard capacitating medium. These data indicate an atypical relationship among cholesterol efflux, ROS production, and PY in equine sperm. Our findings may help to elucidate factors affecting failure of equine IVF under standard conditions.


Asunto(s)
Bicarbonatos/farmacología , Calcio/farmacología , Albúmina Sérica Bovina/farmacología , Capacitación Espermática/efectos de los fármacos , Espermatozoides/efectos de los fármacos , Animales , Tampones (Química) , Bovinos , Colesterol/metabolismo , Femenino , Caballos , Técnicas para Inmunoenzimas , Masculino , Oocitos/citología , Oocitos/efectos de los fármacos , Oocitos/metabolismo , Fosforilación/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Espermatozoides/citología , Espermatozoides/metabolismo , Tirosina/metabolismo
6.
Reprod Domest Anim ; 50(6): 1039-46, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26482800

RESUMEN

Equine in vitro fertilization (IVF) is still inconsistent. In the present work, we studied how modified Whitten's (MW) medium and Tissue Culture Medium 199 (TCM) added with Foetal Bovine Serum (FBS; 10% v/v) or Bovine Serum Albumin (BSA; 7 mg/ml) affected equine gametes to subsequently run IVF trials. Compact (Cp) and expanded (Ex) cumuli equine oocytes were matured and placed in TCM or MW supplemented with BSA or FBS for 18-20 h (no sperm added). In Ex oocytes, TCM-199 added with FBS or BSA resulted in higher metaphase II (MII) rates (75.7% and 62.7%, respectively) than MW added with BSA (54%) or FBS (52.2%; p < 0.05); this was not observed for Cp oocytes. Equine sperm were capacitated in the same media at 10 × 10(6) sperm/ml for 4 h at 37°C; total motility and protein tyrosine phosphorylation (PY) were evaluated. While motility remained unchanged, TCM or MW added with FBS enhanced the number of sperm showing PY-stained tails (25 ± 4.8% and 31 ± 6.6%; mean ± SEM, respectively) over BSA supplemented media (3 ± 1.2% and 11.7 ± 1.1%) for TCM and MW (p < 0.05). In view of the previous results, sperm were capacitated in TCM + FBS and MW + BSA (control); IVF trials were run in the same media supplemented with 200 ng/ml of progesterone, but no fertilization occurred. Our results show that TCM + FBS enhances Ex equine oocyte's meiotic competence over MW + BSA and TCM or MW added with FBS successfully induce equine PY over media supplemented with BSA.


Asunto(s)
Medios de Cultivo/farmacología , Fertilización In Vitro/veterinaria , Caballos , Oocitos/efectos de los fármacos , Proteínas/farmacología , Espermatozoides/efectos de los fármacos , Animales , Femenino , Masculino , Capacitación Espermática/efectos de los fármacos
7.
Reproduction ; 144(4): 411-22, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22843772

RESUMEN

The mechanisms leading to capacitation in stallion sperm are poorly understood. The objective of our study was to define factors associated with regulation of protein tyrosine phosphorylation in stallion sperm. Stallion sperm were incubated for 4 h in modified Whitten's media with or without bicarbonate, calcium, or BSA. When sperm were incubated in air at 30×106/ml at initial pH 7.25, protein tyrosine phosphorylation was detected only in medium containing 25 mM bicarbonate alone; calcium and BSA inhibited phosphorylation. Surprisingly, this inhibition did not occur when sperm were incubated at 10×106/ml. The final pH values after incubation at 30×106 and 10×106 sperm/ml were 7.43 ± 0.04 and 7.83 ± 0.07 (mean ± s.e.m.) respectively. Sperm were then incubated at initial pH values of 7.25, 7.90, or 8.50 in either air or 5% CO2. Protein tyrosine phosphorylation increased with increasing final medium pH, regardless of the addition of bicarbonate or BSA. An increase in environmental pH was observed when raw semen was instilled into the uteri of estrous mares and retrieved after 30 min (from 7.47 ± 0.10 to 7.85 ± 0.08), demonstrating a potential physiological role for pH regulation of capacitation. Sperm incubated in the presence of the calmodulin (CaM) inhibitor W-7 exhibited a dose-dependent increase in protein tyrosine phosphorylation, suggesting that the inhibitory effect of calcium was CaM mediated. These results show for the first time a major regulatory role of external pH, calcium, and CaM in stallion sperm protein tyrosine phosphorylation.


Asunto(s)
Señalización del Calcio , Calmodulina/metabolismo , Caballos/fisiología , Fosfoproteínas/metabolismo , Capacitación Espermática , Espermatozoides/metabolismo , Tirosina/metabolismo , Animales , Calcio/análisis , Señalización del Calcio/efectos de los fármacos , Proteínas Quinasas Dependientes de Calcio-Calmodulina/farmacología , Calmodulina/antagonistas & inhibidores , Quelantes/farmacología , Ácido Egtácico/farmacología , Inhibidores Enzimáticos , Concentración de Iones de Hidrógeno , Masculino , Fosforilación/efectos de los fármacos , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Semen/química , Capacitación Espermática/efectos de los fármacos , Espermatozoides/efectos de los fármacos , Sulfonamidas/farmacología
8.
Neurologia (Engl Ed) ; 36(9): 692-697, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34752347

RESUMEN

OBJECTIVES: The Migraine Disability Assessment (MIDAS) questionnaire is the most frequently used instrument for assessing the level of disability in studies into migraine. This study aims to determine the level of completion of the questionnaire, assess the ease of use, and understand patients' subjective perception of the questionnaire's actual ability to measure disability. MATERIAL AND METHODS: We performed a prospective study of a sample of 78 patients with chronic migraine, determining their level of education and employment status. In a baseline visit, patients were trained to properly complete the questionnaire. At 3 months, we determined the total score and level of completion. Patients also completed a survey measuring ease of use of the questionnaire and patients' perception of whether the score accurately reflected their disability. RESULTS: Only 46% of patients fully completed the questionnaire. Sixty-nine percent reported finding it difficult to complete (this was influenced by patient's employment status but not by educational level). Sixty-two percent of respondents believed that the questionnaire did not fully reflect their own perception of their disability. CONCLUSIONS: Although the validity and consistence of the MIDAS questionnaire are well documented, a high percentage of the study population reported finding it difficult to complete; many patients also considered that the questionnaire did not accurately reflect their disability. Understanding patients' opinions of the suitability of questionnaires used in consultation is crucial to improving completion.


Asunto(s)
Evaluación de la Discapacidad , Trastornos Migrañosos , Humanos , Trastornos Migrañosos/diagnóstico , Percepción , Estudios Prospectivos , Encuestas y Cuestionarios
9.
Opt Lett ; 35(11): 1762-4, 2010 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-20517408

RESUMEN

We demonstrate a method to calibrate a Shack-Hartmann sensor as an orthographic camera. This calibration method permits us to obtain the distance, the rotation matrix between the microlens array and CCD imaging planes, and the projection matrix, which models the projection of the incoming rays to the CCD imaging plane. The proposed calibration method introduces a very compact matrix notation and allows wavefront reconstruction without an explicit centroid search between the reference and distorted spot diagrams. We show a set of simulations in code V that prove the effectiveness of the proposed method.

10.
Biol Reprod ; 81(6): 1106-11, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19605785

RESUMEN

The ability of stallion spermatozoa to produce nitric oxide (NO) before (fresh) and after freezing and thawing (FT) was evaluated by means of flow cytometry after loading the sperm suspension with the probe, 4,5-diaminofluorescenin diacetate. The presence of NO synthase (NOS) was investigated by Western blotting using anti-NOS1, anti-NOS3, or anti-universal NOS antibodies (Abs). While NO was detected both in fresh and FT sperm suspensions, its production increased after cryopreservation only when egg yolk was removed from the extender. Anti-NOS1 Ab intensively labeled a single band with an apparent molecular mass of approximately 83 kDa. On the other hand, the Ab developed against the NOS3 showed a band of approximately 96 kDa in fresh and FT sperm lysates. NO production was positively correlated with sperm motility and velocity after thaw, suggesting an NO role for the functionality of cryopreserved stallion spermatozoa; but the production of NO is compromised in egg yolk-containing extenders.


Asunto(s)
Criopreservación , Óxido Nítrico/metabolismo , Espermatozoides/metabolismo , Animales , Western Blotting , Crioprotectores/farmacología , Yema de Huevo , Citometría de Flujo , Caballos/metabolismo , Masculino , Microscopía Confocal , Óxido Nítrico Sintasa/metabolismo , Semen/metabolismo , Preservación de Semen , Motilidad Espermática , Espermatozoides/efectos de los fármacos , Estadísticas no Paramétricas
11.
Reproduction ; 138(1): 55-63, 2009 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19380427

RESUMEN

Lipid peroxidation (LPO) of stallion spermatozoa was assessed in fresh semen and in samples of the same ejaculates after freezing and thawing. Particular attention was paid to individual differences in the susceptibility to LPO and its possible relationship with freezability. Innate levels of LPO were very low in fresh spermatozoa but increased after thawing, a change that was largely stallion-dependent. The level of LPO in fresh spermatozoa was not correlated with that of the thawed spermatozoa. Negative correlations existed between LPO and intact membranes post-thaw (r=-0.789, P<0.001), and also between LPO and spermatozoa with high mitochondrial membrane potential (Deltapsim) post-thaw (r=-0.689, P<0.001). LPO was also highly and significantly correlated with caspase activity. The correlation between caspase activity in ethidium positive cells and LPO was r=0.772, P<0.001. This LPO is unlikely to represent, per se, a sign of cryopreservation-induced injury, but it is apparently capable of triggering 'apoptotic-like changes' that could result in the sub-lethal cryodamage often seen among surviving spermatozoa.


Asunto(s)
Apoptosis , Compuestos de Boro , Criopreservación/veterinaria , Colorantes Fluorescentes , Caballos , Peroxidación de Lípido , Preservación de Semen/veterinaria , Espermatozoides/metabolismo , Animales , Radioisótopos de Carbono , Caspasas/metabolismo , Supervivencia Celular , Citometría de Flujo , Glutatión Peroxidasa/metabolismo , Masculino , Potencial de la Membrana Mitocondrial , Microscopía Fluorescente , Preservación de Semen/efectos adversos , Espermatozoides/patología , Superóxido Dismutasa/metabolismo
12.
Reprod Domest Anim ; 44(3): 518-22, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19655428

RESUMEN

In an attempt to evaluate the possible relationship between the microbial flora in the stallion ejaculate and its ability to freeze,three ejaculates from five stallions were frozen using a standard protocol. Before freezing, an aliquot was removed for bacteriological analysis. Bacterial growth was observed in all the ejaculates studied. The isolated microorganisms were:Staphylococcus spp. and Micrococcus spp. (in all the stallions), beta-haemolytic Streptococcus (in stallions 3 and 4), Corynebacterium spp. (in stallions 1, 3-5), Rhodococcus spp. (in stallion number 2), Pseudomonas spp. (in stallion number 1) and Klebsiella spp. (in stallions 1, 3 and 5). The presence and richness of Klebsiella and beta-haemolytic Streptococcus in the ejaculate were related to two sperm variables post-thaw,namely the proportion of dead spermatozoa (ethidium+ cells; r = 0.55, p < 0.05) and the amplitude of lateral displacement of the sperm head (ALH, microm; r = -0.56, p < 0.05), respectively.The degree of growth of Corynebacterium spp. in the ejaculate was positively correlated with the percentage of spermatozoa showing high caspase activity post-thaw(r = 0.62, p < 0.05). The presence and number of colonies of beta-haemolytic Streptococcus were negatively correlated (r = -0.55, p < 0.05) with low sperm caspase activity. It is concluded that the microbial flora of the equine ejaculate maybe responsible for some of the sublethal damage experimented by the spermatozoa during cryopreservation.


Asunto(s)
Criopreservación/veterinaria , Caballos/microbiología , Preservación de Semen/veterinaria , Semen/microbiología , Espermatozoides/fisiología , Animales , Bacterias/crecimiento & desarrollo , Bacterias/aislamiento & purificación , Caspasas/análisis , Corynebacterium/aislamiento & purificación , Citometría de Flujo , Klebsiella/aislamiento & purificación , Masculino , Micrococcus/aislamiento & purificación , Pseudomonas/aislamiento & purificación , Rhodococcus/aislamiento & purificación , Preservación de Semen/métodos , Motilidad Espermática , Staphylococcus/aislamiento & purificación , Streptococcus/aislamiento & purificación
13.
Reprod Domest Anim ; 44(3): 419-23, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19055563

RESUMEN

In an attempt to improve the information obtained after computer-assisted sperm analysis (CASA), data from five stallions (three ejaculates from each) were analysed before (fresh, extended semen) and after cryopreservation using traditional statistics as well as a cluster analysis. The data matrix consisted of 13 987 observations of individual spermatozoa for fresh, extended semen, and 8305 for frozen-thawed samples. As expected, freezing and thawing resulted in a marked decrease of CASA-derived variables of sperm kinematics. All sperm velocities were significantly lower in frozen-thawed samples than in samples before cooling. Using sperm velocities, six sperm subpopulations were identified in fresh semen (S1-S6). As such, subpopulations S1 and S2 were characterized by low sperm velocities, subpopulations S3 and S4 corresponded to spermatozoa depicting medium speed values, and finally, subpopulations S5 and S6 were those depicting the highest velocities. After freezing and thawing, four sperm subpopulations were identified, listed as nr FT1 to FT4. While subpopulations FT1-FT3 were characterized by low sperm velocities, and thus corresponded speed-wise to those listed as S1-S4 for fresh, extended semen, the one called number FT4 in frozen semen was characterized by high velocities, of the same range as that of the subpopulations S5 and S6 for fresh spermatozoa. The sperm subpopulation structure varied among stallions, but the cluster analysis hereby assayed was able to provide valuable information about the freezability of the samples that the customary statistics did not reveal.


Asunto(s)
Criopreservación/veterinaria , Caballos , Preservación de Semen/veterinaria , Espermatozoides/fisiología , Animales , Calor , Masculino , Motilidad Espermática/fisiología , Espermatozoides/citología
14.
Reprod Domest Anim ; 44(2): 345-9, 2009 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19144010

RESUMEN

While, for a long time, the role of mitochondria in sperm physiology and pathology has been largely ignored, recent research points out the mitochondria as a major organelle with key roles in sperm function both under physiological and biotechnological conditions. This paper briefly reviews these novel findings regarding the role of mitochondria in sperm, paying special attention to the most practical, readily applicable, aspects of the topic such as their role as a major source of the sublethal damage that sperm experiments after cryopreservation.


Asunto(s)
Mitocondrias/fisiología , Espermatozoides/anomalías , Espermatozoides/fisiología , Animales , Muerte Celular/fisiología , Criopreservación/veterinaria , Epidídimo/citología , Fertilización/fisiología , Masculino , Preservación de Semen/efectos adversos , Preservación de Semen/veterinaria , Motilidad Espermática/fisiología , Espermatozoides/ultraestructura
15.
Reprod Domest Anim ; 44(3): 523-6, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-18992085

RESUMEN

The present study attempted to select the subpopulation of stallion spermatozoa that best survived a conventional freezing and thawing procedure, using centrifugation of post-thawed semen samples through a single layer of a glycidoxypropyltrimethoxysilane-coated silica colloid with a species-specific formulation (Androcoll-E). After freezing and thawing, four sperm subpopulations were identified, listed as FT1 to FT4. While subpopulations FT1 and FT2 were characterized by low sperm velocity, high velocities characterized the ones called FT3 and FT4. The single-layer centrifugation (SLC)-handled sperm sample was enriched in subpopulation FT3, reaching a proportion of 82.6% of the present spermatozoa, in contrast with the non-filtered control post-thawed semen, where this sperm subpopulation only accounted for 16.3% of the total. It is concluded that in the equine industry, the SLC is a practical, easy-to-perform approach to improve the quality of equine frozen-thawed semen samples.


Asunto(s)
Centrifugación/veterinaria , Coloides , Criopreservación/veterinaria , Caballos , Preservación de Semen/veterinaria , Espermatozoides/citología , Animales , Separación Celular/métodos , Separación Celular/veterinaria , Supervivencia Celular , Centrifugación/métodos , Calor , Masculino , Preservación de Semen/métodos , Dióxido de Silicio , Motilidad Espermática , Espermatozoides/fisiología
16.
J Androl ; 29(2): 213-21, 2008.
Artículo en Inglés | MEDLINE | ID: mdl-17978341

RESUMEN

The kinematics of the appearance of apoptotic markers was studied by flow cytometry and immunoblot assays in equine spermatozoa subjected to freezing and thawing. Caspase activity, low mitochondrial membrane potential, and increases in sperm membrane permeability were observed in all of the phases of the cryopreservation procedure. Freezing and thawing caused an increase in membrane permeability and changes in the pattern of caspase activity; decreases in mitochondrial membrane potential were observed after centrifugation and cooling to 4 degrees C and after freezing and thawing. It is proposed that sperm mitochondria may be directly involved in the subtle damage that is present in most spermatozoa surviving freezing and thawing.


Asunto(s)
Criopreservación/veterinaria , Preservación de Semen/veterinaria , Espermatozoides/citología , Animales , Apoptosis , Bencimidazoles/farmacología , Carbocianinas/farmacología , Caspasas/metabolismo , Permeabilidad de la Membrana Celular , Citometría de Flujo , Caballos , Masculino , Potencial de la Membrana Mitocondrial , Espermatozoides/efectos de los fármacos
17.
Neurologia (Engl Ed) ; 2018 Nov 16.
Artículo en Inglés, Español | MEDLINE | ID: mdl-30455123

RESUMEN

OBJECTIVES: The Migraine Disability Assessment (MIDAS) questionnaire is the most frequently used instrument for assessing the level of disability in studies into migraine. This study aims to determine the level of completion of the questionnaire, assess the ease of use, and understand patients' subjective perception of the questionnaire's actual ability to measure disability. MATERIAL AND METHODS: We performed a prospective study of a sample of 78 patients with chronic migraine, determining their level of education and employment status. In a baseline visit, patients were trained to properly complete the questionnaire. At 3 months, we determined the total score and level of completion. Patients also completed a survey measuring ease of use of the questionnaire and patients' perception of whether the score accurately reflected their disability. RESULTS: Only 46% of patients fully completed the questionnaire. Sixty-nine percent reported finding it difficult to complete (this was influenced by patient's employment status but not by educational level). Sixty-two percent of respondents believed that the questionnaire did not fully reflect their own perception of their disability. CONCLUSIONS: Although the validity and consistence of the MIDAS questionnaire are well documented, a high percentage of the study population reported finding it difficult to complete; many patients also considered that the questionnaire did not accurately reflect their disability. Understanding patients' opinions of the suitability of questionnaires used in consultation is crucial to improving completion.

18.
Anim Reprod Sci ; 187: 13-19, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29032866

RESUMEN

Heat Shock Proteins (HSP) is a family of proteins that protects cells from high temperatures. The present work aimed to elucidate the role that HSP90 exerts on boar sperm incubated under heat stress conditions on viability, total motility (TM), progressive motility (PM), acrosome status, mitochondrial membrane potential and plasma membrane lipid organization. Sperm were incubated in non-capacitating conditions (Tyrode's basal medium or TBM) for 3, 8 and 24h or in capacitating conditions (Tyrode's complete medium or TCM) for 4h at 38.5°C or 40°C (Heat stress) in the presence or absence of 5 or 20µM of 17-AAG, a specific HSP90 inhibitor. Sperm viability was not affected by the presence of 17-AAG in any condition tested compared with its own control (at the same temperature and incubation time). In non-capacitating conditions TM (22.7±4.1 vs. 1.9±1.1; % mean±SEM), PM (3.1±0.9 vs. 0) and high mitochondrial membrane potential (19.5±2.2 vs. 11.8±0.8) decreased significantly in sperm incubated at 40°C for 24h in the presence of 20µM 17-AAG (control vs. 20µM 17-AAG, respectively; p<0.05). In sperm incubated at 38.5°C only a mild decrease in TM was observed (48.7±3.1 vs. 32.1±4.8; control vs. 20µM 17-AAG, respectively; p<0.05). However, under capacitating conditions none of the sperm parameters studied were affected by 17-AAG after 4h of incubation. These results demonstrate for the first time the role of HSP90 in the maintenance of boar sperm motility and mitochondrial membrane potential during prolonged heat stress in non-capacitating conditions.


Asunto(s)
Proteínas HSP90 de Choque Térmico/metabolismo , Potencial de la Membrana Mitocondrial , Motilidad Espermática , Espermatozoides/fisiología , Animales , Calor , Masculino , Espermatozoides/citología , Estrés Fisiológico , Porcinos
19.
Andrology ; 5(6): 1131-1140, 2017 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-28980456

RESUMEN

AMP-activated kinase (AMPK) plays a key function in maintaining cellular energy homeostasis. We recently identified and localized AMPK protein in human spermatozoa and showed that inhibition of AMPK activity significantly modified human sperm motility. Recently, AMPK has gained great relevance as prime target for pharmacological approaches in several energy-related pathologies and therefore pharmacological research is aimed to develop direct AMPK-activating compounds such as A769662. Our aim was to investigate the effect of A769662 in essential functional processes of human spermatozoa. Human spermatozoa were incubated in the presence or absence of the AMPK activator A769662 for different incubation times (0-20 h) and motility was evaluated by CASA system whereas other functional parameters were evaluated by flow cytometry. A769662 treatment significantly reduces the percentages of motile, progressive, and rapid spermatozoa starting at 2 h. Moreover, AMPK activator in human spermatozoa causes a significant reduction in any velocity measured, which is concomitant to a significant decrease in the percentage of rapid spermatozoa, both at short- (2-3 h) and long-time treatment (20 h). Treatment of human spermatozoa with A769662 does not significantly alter any of the following functional parameters: sperm viability, mitochondrial membrane potential, phosphatidylserine translocation to the outer leaf of plasma membrane, acrosome membrane integrity, or mitochondrial superoxide anion production. In summary, our results suggest that AMPK in human spermatozoa contributes to the regulation of sperm motility, without affecting basic physiological parameters of human spermatozoa (viability, mitochondrial membrane potential or reactive oxygen species production, acrosome membrane integrity, phosphatidylserine exposure at plasma membrane). As sperm motility is required in the female reproductive tract to achieve fertilization, we conclude that AMPK is an essential regulatory kinase of the human spermatozoa function. This conclusion needs to be taken into account when AMPK is elected as prime target in pharmacological approaches for several energy-related pathologies.


Asunto(s)
Pironas/farmacología , Motilidad Espermática/efectos de los fármacos , Espermatozoides/efectos de los fármacos , Tiofenos/farmacología , Proteínas Quinasas Activadas por AMP/efectos de los fármacos , Compuestos de Bifenilo , Regulación hacia Abajo , Humanos , Masculino , Espermatozoides/enzimología
20.
Neurología (Barc., Ed. impr.) ; 36(9): 692-697, noviembre-diciembre 2021. tab, graf
Artículo en Español | IBECS (España) | ID: ibc-220132

RESUMEN

Objetivos: El cuestionario Migraine Disability Assessment (MIDAS) es el instrumento más empleado para valorar el grado de discapacidad en los estudios de migraña. El objetivo del estudio es determinar el nivel de cumplimentación del cuestionario, valorar su facilidad de uso y conocer la percepción subjetiva del paciente sobre la capacidad del cuestionario para medir realmente su discapacidad.Material y métodosEstudio prospectivo sobre una población de 78 pacientes con migraña crónica. Se determina el nivel educativo y la situación laboral. En la visita basal se adiestra a los pacientes sobre la correcta cumplimentación del cuestionario. A los 3 meses se determina la puntuación total y el nivel de cumplimentación. Además los pacientes contestan una encuesta que mide: facilidad de uso y percepción del paciente sobre si la escala refleja su propia discapacidad.ResultadosSolo el 46% rellena completamente el cuestionario. El 69% de los pacientes indica que el cuestionario no les resulta fácil de cumplimentar (resultado no influido por el nivel educativo, pero sí por la situación laboral de los pacientes). El 62% de los encuestados opina que el cuestionario no refleja completamente su propia percepción de discapacidad.ConclusionesAunque está más que demostrada la validez y consistencia del cuestionario MIDAS, un porcentaje elevado de nuestra población reconoce que el cuestionario no es fácil de rellenar y además es percibido por muchos de nuestros pacientes como un cuestionario que no refleja adecuadamente su discapacidad. Conocer la opinión de los pacientes sobre la idoneidad de los cuestionarios administrados en las consultas es crucial para mejorar su cumplimentación. (AU)


Objectives: The Migraine Disability Assessment (MIDAS) questionnaire is the most frequently used instrument for assessing the level of disability in studies into migraine. This study aims to determine the level of completion of the questionnaire, assess the ease of use, and understand patients’ subjective perception of the questionnaire's actual ability to measure disability.Material and methodsWe performed a prospective study of a sample of 78 patients with chronic migraine, determining their level of education and employment status. In a baseline visit, patients were trained to properly complete the questionnaire. At 3 months, we determined the total score and level of completion. Patients also completed a survey measuring ease of use of the questionnaire and patients’ perception of whether the score accurately reflected their disability.ResultsOnly 46% of patients fully completed the questionnaire. Sixty-nine percent reported finding it difficult to complete (this was influenced by patient's employment status but not by educational level). Sixty-two percent of respondents believed that the questionnaire did not fully reflect their own perception of their disability.ConclusionsAlthough the validity and consistence of the MIDAS questionnaire are well documented, a high percentage of the study population reported finding it difficult to complete; many patients also considered that the questionnaire did not accurately reflect their disability. Understanding patients’ opinions of the suitability of questionnaires used in consultation is crucial to improving completion. (AU)


Asunto(s)
Humanos , Trastornos Migrañosos/diagnóstico , Percepción Auditiva , Cefalea , Estudios Prospectivos , Encuestas y Cuestionarios
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