Cultured rat hippocampal neurons with short processes were investigated using the whole cell volt... more Cultured rat hippocampal neurons with short processes were investigated using the whole cell voltage clamp under conditions appropriate for isolating Na+ currents. After incubation of the neuron culture for a period of 15–30 min in 1 mM sodium valproate, several parameters of the Na+ current were changed. The peak Na+ conductance g p, measured using hyperpolarizing prepulses, was reduced by valproate in a voltage-dependent manner. In the membrane voltage range from -30 to +20 mV, this reduction showed a linear dependence on voltage, increasing from about zero to approximately 30% of g p, the maximum peak Na+ conductance of the neuron. At the holding voltage of -70 mV, the inactivation parameter h t8 decreased from 0.88 in the control to 0.64 in the valproate solution. This reduction originated mainly from a 10 mV shift in the sigmoid relation between h t8 and membrane voltage along the voltage axis to hyperpolarizing potentials. The decay of the maximum peak Na+ current (inactivation) could be fitted by a biexponential function. Time constants of the fast and slow component at -20 mV decreased in valproate by about 50%. Valproate also retarded the recovery from inactivation, as determined at the holding voltage. The sigmoid recovery from inactivation could reasonably be described by an exponential function with time constant τ r and delay time Δt. Both τ r and At increased more than 200% in valproate. Our results indicate that valproate affected the Na+ current in hippocampal neurons in a way that contributed to a considerable depression of Na+ reactivation. This explains the frequency-dependent inhibition of action potentials as observed in mammalian central nervous tissue and may be the principal action of the anticonvulsant.
L'A, studia in questa Memoria la distribuzione delle tensioni interne in un solido elastico conve... more L'A, studia in questa Memoria la distribuzione delle tensioni interne in un solido elastico convesso quando gli spostamenti, dati in superficie, sono diversi da zero solo in una piccola zona. Nei campi a due dimensioni il risultato è esteso al caso in cui sul contorno siano date le forze; si giunge così ad una dimostrazione del principio del De Saint Venant.
The authors studied hyperactivated motility of human spermatozoa as a method of evaluating capaci... more The authors studied hyperactivated motility of human spermatozoa as a method of evaluating capacitation by examining its relationship to results of zona-free hamster oocyte sperm penetration assays (SPA) of semen samples from 50 men attending the infertility clinic. Hyperactivated motility was assessed in the seminal plasma and after swim-up preparation of spermatozoa at 1, 3, and 24 hours of incubation in capacitation media using a computer-assisted semen analysis system equipped with a hyperactivation module. Hyperactivated motility reached a peak at 1 hour and plateaued at 3 hours. The percentage of spermatozoa in seminal plasma with star-spin hyperactivated motility was significantly lower in the group showing no penetration in the SPA. The hyperactivated motility characteristics did not differ in the groups with positive or negative penetration. Correlation analysis failed to show any significant relationship between the hyperactivated motility parameters and SPA score. When the hyperactivated motility characteristics were compared in samples with normal and abnormal semen analyses, the total percentage of spermatozoa with hyperactivated motility and the percentage with star-spin at 3 hours were significantly lower in the group with abnormal semen analysis. The data indicate that lower hyperactivated motility of spermatozoa was found in patients with a score of zero for SPA and in patients with abnormal semen analysis. It was concluded that although no direct correlations were found between the results of SPA and hyperactivated motility, evaluating hyperactivated motility may still be useful as an early indicator of capacitation abnormalities of human spermatozoa not measured by SPA.
Revista Brasileira De Zootecnia-brazilian Journal of Animal Science, 2003
RESUMO -Os valores econômicos para características de gado de leite foram calculados a partir de ... more RESUMO -Os valores econômicos para características de gado de leite foram calculados a partir de dados econômicos e zootécnicos do Sistema de Produção de Leite a Pasto com Gado Mestiço da Embrapa Gado de Leite. Estes valores foram estimados por dois métodos: 1) utilizando a derivada parcial do lucro com respeito a cada característica e 2) pela derivada parcial da razão receita/custo, para cada característica, ambas as derivadas avaliadas no valor médio das outras características. Com este último método, os valores econômicos das características consideradas, expressos em equivalente kg de leite, foram: veículo (leite sem gordura e proteína), 0,77 /kg, gordura, -5,0 /kg, proteína -3,65 /kg, mamite, -155,34 /caso, fluxo lácteo, 146,00 /kg/min, número de serviços por concepção, -104,5 /serviço, idade aos 330 kg, -1,27 /dia, dias desde os 330 kg até o primeiro parto, -0,51 /dia, vida útil, 49,63 /ano e peso da vaca seca, -4,34 /kg. Os valores obtidos com o método 1 foram semelhantes. Os resultados confirmam comunicações da literatura de valor econômico negativo para a gordura e proteína do leite quando estes componentes não são remunerados, de maior valor econômico relativo para a redução do peso corporal das vacas que para o aumento da produção de leite, da grande importância da mamite, fluxo lácteo e vida útil e da pouca importância relativa do intervalo em dias desde os 330 kg até o primeiro parto.
In the central north of Chile (25°00'S-32°10'S) an anchoveta fishery (Engraulis ringens) operates... more In the central north of Chile (25°00'S-32°10'S) an anchoveta fishery (Engraulis ringens) operates on a small scale. In spite of being controlled by total allowed catches, the life history parameters used to estimate its abundance remain unknown. We estimated the basic biological parameters as growth, maturity and natural mortality for anchovy inhabiting the Chilean central north. The von Bertalanffy parameters were an annual growth rate (k) of 0.91 year -1 , asymptotic length (L ∞ ) at 18.72 cm, and age at zero length (t o ) of -0.01 year. The maximum value of the gonadosomatic index in September and the high frequency of mature stages between July and December, support that the main spawning period of anchoveta in the area would occur from August to October of each year. The mean maturity length (L m50% ) was estimated at 12.5 cm and the natural mortality (M) was 1.30 year -1 . The biological parameters of E. ringens located in central north of Chile are in concordance with the life history characteristics of small pelagic fish.
Current induced in cultured cerebellar granule cells by the bath application of kainate (500 μM) ... more Current induced in cultured cerebellar granule cells by the bath application of kainate (500 μM) was measured using the conventional patch-clamp technique. Two different kinds of responses were observed after the agonits perfusion. Some cells exhibited small inward whole-cell currents: 116±40 pA (7 cells) at a clamp potential of−60 mV; in other cells, the agonist induced significantly larger currents: 420±35 pA (6 cells) at a clamp potential of −60 mV. The current flowing in the agonist-activated ionic channels was indirectly estimated by processing the fluctuations of whole-cell current by means of an original parametric method. Mean conductance of the underlying channels was then determined from the single-channel current estimated at different clamp potentials. In the cells exhibiting small inward currents, the mean conductance was equal to 0.5±0.2 pS (7 cells), whereas in the cells with large inward currents it was 3±0.4 pS (6 cells). This result gives a coherent explanation of the different kinds of responses observed at macroscopic level in the whole-cell current and confirms that kainate-activated channels can exhibit different levels of conductance.
Several Mycoplasma species may adversely affect bovine spermatozoa viability and embryo developme... more Several Mycoplasma species may adversely affect bovine spermatozoa viability and embryo development. Mycoplasma mycoides ssp. mycoides large-colony (LC) has been isolated from naturally aborted bovine fetuses and from bull semen. The objective of this study was to evaluate whether M. mycoides ssp. mycoides LC contaminated bovine ejaculates could (i) impair in vitro fertilizing ability of bull spermatozoa, (ii) impair embryo development, and (iii) evaluate potential spread by reproductive technologies. In the present study, spermatozoa of 10 fertile bulls were contaminated with M. mycoides ssp. mycoides LC, at a final concentration of 1.5 million CFU/ml and incubated for 60 min before evaluating spermatozoa motility and acrosome reaction inducibility with calcium ionophore. In addition, in vitro contaminated semen of a bull previously shown to have a good in vitro fertilizing ability, was used in an IVF procedure. Embryo development stage on Day-7 of culture was evaluated. Spermatozoa and embryos at morula and blastocyst stages were routinely processed for transmission electron microscopy observation. Both mean total and progressive motility decreased (P < 0.01 ) upon spermatozoa incubation with Mycoplasma. One-hour incubation with calcium ionophore increased the percentage of acrosome-reacted spermatozoa, although Mycoplasma contamination reduced calcium ionophore treatment efficacy (P < 0.05). Ultrastructurally, Mycoplasma microorganisms appeared as moderately electron-dense sphere-shaped particles, adhering to cell ଝ Bovine spermatozoa in vitro contaminated with M. mycoides ssp mycoides LC L. Sylla et al. / Animal Reproduction Science 85 (2005) 81-93 membranes. Sperm mid-piece sections showed numeric aberrations of the central singlets such as nine + zero or nine + one of the axonemal complex. Further morphological abnormalities included partial or total absence of dinein arms and radial fibers, with lack of the bridge and the central ring in 35.00 ± 4.20% of contaminated cells, whereas these abnormalities were not observed in uninfected ones. The IVF trials showed that two-four cell blocks were higher (P < 0.05) in the infected group. Ultrastructure of Day-7 contaminated embryos showed Mycoplasma particles adhering and infiltrating the outer layer of the zona pellucida. Our investigations suggest that M. mycoides ssp. mycoides LC contaminating the bovine ejaculate induced adverse effects on in vitro spermatozoa-fertilizing ability and embryonic development. Some satisfactory quality transferable embryos could be produced in contaminated IVF systems. This could imply a potential transmission of this microorganism through reproductive technologies.
Journal of Assisted Reproduction and Genetics, 2002
Purpose : The study aimed to evaluate the relationship between the zona pellucida induced acrosom... more Purpose : The study aimed to evaluate the relationship between the zona pellucida induced acrosome reaction (ZIAR) and (i) percentage normal spermatozoa as well as (ii) sperm–zona pellucida binding potential among men referred for a routine semen analysis. Methods : Semen samples of 164 consecutive men referred to the andrology laboratory for routine semen analysis were studied. Semen samples were analyzed using the new WHO standards (strict criteria). ZIAR was recorded with a lectin conjugated Pisum sativum agglutinin microassay, while sperm–zona binding was evaluated with a standard hemizona assay (HZA). Results : Andrology patients were divided according to the percentage normal spermatozoa in the ejaculate, namely n = 71), 5–14% normal forms (n = 73), and >14% normal forms (n = 20). ZIAR data of the 14% groups was (9.6 ± 0.6)%, (13.9 ± 0.5)%, and (15.0 ± 1.1)%, respectively. The ZIAR data of fertile control men was (26.6 ± 1.4)% which differed significantly from the three andrology referrals groups. Likewise significant differences were recorded during the hemizona assay namely, 38.0% (14% normal forms). Among the group with >14% normal forms, five cases had impaired ZIAR outcome ( Conclusions : ZIAR testing should become part of the second level of male fertility investigations, i.e., sperm functional testing, since 15% of andrology referrals revealed an impaired acrosome reaction response to solubilized zona pellucida.
Florencia Caquetá, Marzo 28 de 2014 Señor SAUL BUITRAGO PEREZ Jefe Oficina de Tributo, alcaldía d... more Florencia Caquetá, Marzo 28 de 2014 Señor SAUL BUITRAGO PEREZ Jefe Oficina de Tributo, alcaldía de Florencia Ciudad. Asunto: CANCELACION MATRICULA DE INDUSTRIA Y COMERCIO. Yo, FABIO GRANADA OCAMPO identificado con cedula de ciudadanía, 1.284.609 de Filadelfia Caldas, le hago llegar el formulario de cancelación del establecimiento comercial que se encontraba a mi nombre (chatarrería el paisa), esto debido a que fue vendido a la señora LUZ AMPARO GRANADA
Cultured rat hippocampal neurons with short processes were investigated using the whole cell volt... more Cultured rat hippocampal neurons with short processes were investigated using the whole cell voltage clamp under conditions appropriate for isolating Na+ currents. After incubation of the neuron culture for a period of 15–30 min in 1 mM sodium valproate, several parameters of the Na+ current were changed. The peak Na+ conductance g p, measured using hyperpolarizing prepulses, was reduced by valproate in a voltage-dependent manner. In the membrane voltage range from -30 to +20 mV, this reduction showed a linear dependence on voltage, increasing from about zero to approximately 30% of g p, the maximum peak Na+ conductance of the neuron. At the holding voltage of -70 mV, the inactivation parameter h t8 decreased from 0.88 in the control to 0.64 in the valproate solution. This reduction originated mainly from a 10 mV shift in the sigmoid relation between h t8 and membrane voltage along the voltage axis to hyperpolarizing potentials. The decay of the maximum peak Na+ current (inactivation) could be fitted by a biexponential function. Time constants of the fast and slow component at -20 mV decreased in valproate by about 50%. Valproate also retarded the recovery from inactivation, as determined at the holding voltage. The sigmoid recovery from inactivation could reasonably be described by an exponential function with time constant τ r and delay time Δt. Both τ r and At increased more than 200% in valproate. Our results indicate that valproate affected the Na+ current in hippocampal neurons in a way that contributed to a considerable depression of Na+ reactivation. This explains the frequency-dependent inhibition of action potentials as observed in mammalian central nervous tissue and may be the principal action of the anticonvulsant.
L'A, studia in questa Memoria la distribuzione delle tensioni interne in un solido elastico conve... more L'A, studia in questa Memoria la distribuzione delle tensioni interne in un solido elastico convesso quando gli spostamenti, dati in superficie, sono diversi da zero solo in una piccola zona. Nei campi a due dimensioni il risultato è esteso al caso in cui sul contorno siano date le forze; si giunge così ad una dimostrazione del principio del De Saint Venant.
The authors studied hyperactivated motility of human spermatozoa as a method of evaluating capaci... more The authors studied hyperactivated motility of human spermatozoa as a method of evaluating capacitation by examining its relationship to results of zona-free hamster oocyte sperm penetration assays (SPA) of semen samples from 50 men attending the infertility clinic. Hyperactivated motility was assessed in the seminal plasma and after swim-up preparation of spermatozoa at 1, 3, and 24 hours of incubation in capacitation media using a computer-assisted semen analysis system equipped with a hyperactivation module. Hyperactivated motility reached a peak at 1 hour and plateaued at 3 hours. The percentage of spermatozoa in seminal plasma with star-spin hyperactivated motility was significantly lower in the group showing no penetration in the SPA. The hyperactivated motility characteristics did not differ in the groups with positive or negative penetration. Correlation analysis failed to show any significant relationship between the hyperactivated motility parameters and SPA score. When the hyperactivated motility characteristics were compared in samples with normal and abnormal semen analyses, the total percentage of spermatozoa with hyperactivated motility and the percentage with star-spin at 3 hours were significantly lower in the group with abnormal semen analysis. The data indicate that lower hyperactivated motility of spermatozoa was found in patients with a score of zero for SPA and in patients with abnormal semen analysis. It was concluded that although no direct correlations were found between the results of SPA and hyperactivated motility, evaluating hyperactivated motility may still be useful as an early indicator of capacitation abnormalities of human spermatozoa not measured by SPA.
Revista Brasileira De Zootecnia-brazilian Journal of Animal Science, 2003
RESUMO -Os valores econômicos para características de gado de leite foram calculados a partir de ... more RESUMO -Os valores econômicos para características de gado de leite foram calculados a partir de dados econômicos e zootécnicos do Sistema de Produção de Leite a Pasto com Gado Mestiço da Embrapa Gado de Leite. Estes valores foram estimados por dois métodos: 1) utilizando a derivada parcial do lucro com respeito a cada característica e 2) pela derivada parcial da razão receita/custo, para cada característica, ambas as derivadas avaliadas no valor médio das outras características. Com este último método, os valores econômicos das características consideradas, expressos em equivalente kg de leite, foram: veículo (leite sem gordura e proteína), 0,77 /kg, gordura, -5,0 /kg, proteína -3,65 /kg, mamite, -155,34 /caso, fluxo lácteo, 146,00 /kg/min, número de serviços por concepção, -104,5 /serviço, idade aos 330 kg, -1,27 /dia, dias desde os 330 kg até o primeiro parto, -0,51 /dia, vida útil, 49,63 /ano e peso da vaca seca, -4,34 /kg. Os valores obtidos com o método 1 foram semelhantes. Os resultados confirmam comunicações da literatura de valor econômico negativo para a gordura e proteína do leite quando estes componentes não são remunerados, de maior valor econômico relativo para a redução do peso corporal das vacas que para o aumento da produção de leite, da grande importância da mamite, fluxo lácteo e vida útil e da pouca importância relativa do intervalo em dias desde os 330 kg até o primeiro parto.
In the central north of Chile (25°00'S-32°10'S) an anchoveta fishery (Engraulis ringens) operates... more In the central north of Chile (25°00'S-32°10'S) an anchoveta fishery (Engraulis ringens) operates on a small scale. In spite of being controlled by total allowed catches, the life history parameters used to estimate its abundance remain unknown. We estimated the basic biological parameters as growth, maturity and natural mortality for anchovy inhabiting the Chilean central north. The von Bertalanffy parameters were an annual growth rate (k) of 0.91 year -1 , asymptotic length (L ∞ ) at 18.72 cm, and age at zero length (t o ) of -0.01 year. The maximum value of the gonadosomatic index in September and the high frequency of mature stages between July and December, support that the main spawning period of anchoveta in the area would occur from August to October of each year. The mean maturity length (L m50% ) was estimated at 12.5 cm and the natural mortality (M) was 1.30 year -1 . The biological parameters of E. ringens located in central north of Chile are in concordance with the life history characteristics of small pelagic fish.
Current induced in cultured cerebellar granule cells by the bath application of kainate (500 μM) ... more Current induced in cultured cerebellar granule cells by the bath application of kainate (500 μM) was measured using the conventional patch-clamp technique. Two different kinds of responses were observed after the agonits perfusion. Some cells exhibited small inward whole-cell currents: 116±40 pA (7 cells) at a clamp potential of−60 mV; in other cells, the agonist induced significantly larger currents: 420±35 pA (6 cells) at a clamp potential of −60 mV. The current flowing in the agonist-activated ionic channels was indirectly estimated by processing the fluctuations of whole-cell current by means of an original parametric method. Mean conductance of the underlying channels was then determined from the single-channel current estimated at different clamp potentials. In the cells exhibiting small inward currents, the mean conductance was equal to 0.5±0.2 pS (7 cells), whereas in the cells with large inward currents it was 3±0.4 pS (6 cells). This result gives a coherent explanation of the different kinds of responses observed at macroscopic level in the whole-cell current and confirms that kainate-activated channels can exhibit different levels of conductance.
Several Mycoplasma species may adversely affect bovine spermatozoa viability and embryo developme... more Several Mycoplasma species may adversely affect bovine spermatozoa viability and embryo development. Mycoplasma mycoides ssp. mycoides large-colony (LC) has been isolated from naturally aborted bovine fetuses and from bull semen. The objective of this study was to evaluate whether M. mycoides ssp. mycoides LC contaminated bovine ejaculates could (i) impair in vitro fertilizing ability of bull spermatozoa, (ii) impair embryo development, and (iii) evaluate potential spread by reproductive technologies. In the present study, spermatozoa of 10 fertile bulls were contaminated with M. mycoides ssp. mycoides LC, at a final concentration of 1.5 million CFU/ml and incubated for 60 min before evaluating spermatozoa motility and acrosome reaction inducibility with calcium ionophore. In addition, in vitro contaminated semen of a bull previously shown to have a good in vitro fertilizing ability, was used in an IVF procedure. Embryo development stage on Day-7 of culture was evaluated. Spermatozoa and embryos at morula and blastocyst stages were routinely processed for transmission electron microscopy observation. Both mean total and progressive motility decreased (P < 0.01 ) upon spermatozoa incubation with Mycoplasma. One-hour incubation with calcium ionophore increased the percentage of acrosome-reacted spermatozoa, although Mycoplasma contamination reduced calcium ionophore treatment efficacy (P < 0.05). Ultrastructurally, Mycoplasma microorganisms appeared as moderately electron-dense sphere-shaped particles, adhering to cell ଝ Bovine spermatozoa in vitro contaminated with M. mycoides ssp mycoides LC L. Sylla et al. / Animal Reproduction Science 85 (2005) 81-93 membranes. Sperm mid-piece sections showed numeric aberrations of the central singlets such as nine + zero or nine + one of the axonemal complex. Further morphological abnormalities included partial or total absence of dinein arms and radial fibers, with lack of the bridge and the central ring in 35.00 ± 4.20% of contaminated cells, whereas these abnormalities were not observed in uninfected ones. The IVF trials showed that two-four cell blocks were higher (P < 0.05) in the infected group. Ultrastructure of Day-7 contaminated embryos showed Mycoplasma particles adhering and infiltrating the outer layer of the zona pellucida. Our investigations suggest that M. mycoides ssp. mycoides LC contaminating the bovine ejaculate induced adverse effects on in vitro spermatozoa-fertilizing ability and embryonic development. Some satisfactory quality transferable embryos could be produced in contaminated IVF systems. This could imply a potential transmission of this microorganism through reproductive technologies.
Journal of Assisted Reproduction and Genetics, 2002
Purpose : The study aimed to evaluate the relationship between the zona pellucida induced acrosom... more Purpose : The study aimed to evaluate the relationship between the zona pellucida induced acrosome reaction (ZIAR) and (i) percentage normal spermatozoa as well as (ii) sperm–zona pellucida binding potential among men referred for a routine semen analysis. Methods : Semen samples of 164 consecutive men referred to the andrology laboratory for routine semen analysis were studied. Semen samples were analyzed using the new WHO standards (strict criteria). ZIAR was recorded with a lectin conjugated Pisum sativum agglutinin microassay, while sperm–zona binding was evaluated with a standard hemizona assay (HZA). Results : Andrology patients were divided according to the percentage normal spermatozoa in the ejaculate, namely n = 71), 5–14% normal forms (n = 73), and >14% normal forms (n = 20). ZIAR data of the 14% groups was (9.6 ± 0.6)%, (13.9 ± 0.5)%, and (15.0 ± 1.1)%, respectively. The ZIAR data of fertile control men was (26.6 ± 1.4)% which differed significantly from the three andrology referrals groups. Likewise significant differences were recorded during the hemizona assay namely, 38.0% (14% normal forms). Among the group with >14% normal forms, five cases had impaired ZIAR outcome ( Conclusions : ZIAR testing should become part of the second level of male fertility investigations, i.e., sperm functional testing, since 15% of andrology referrals revealed an impaired acrosome reaction response to solubilized zona pellucida.
Florencia Caquetá, Marzo 28 de 2014 Señor SAUL BUITRAGO PEREZ Jefe Oficina de Tributo, alcaldía d... more Florencia Caquetá, Marzo 28 de 2014 Señor SAUL BUITRAGO PEREZ Jefe Oficina de Tributo, alcaldía de Florencia Ciudad. Asunto: CANCELACION MATRICULA DE INDUSTRIA Y COMERCIO. Yo, FABIO GRANADA OCAMPO identificado con cedula de ciudadanía, 1.284.609 de Filadelfia Caldas, le hago llegar el formulario de cancelación del establecimiento comercial que se encontraba a mi nombre (chatarrería el paisa), esto debido a que fue vendido a la señora LUZ AMPARO GRANADA
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