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A Medrano

Publications and source records attributed to A Medrano.

14 recordsLinked to original sources

Sperm motility patterns and metabolism in Catalonian donkey semen.

The Sperm-Class Analyzer detected four subpopulations of spermatozoa with different motility characteristics in the ejaculate of the Catalonian donkey. Significant differences (P < 0.001) in the distribution of these subpopulations, as well as in total sperm number and percentage total motility, were seen in the diluted semen of four sampled donkeys. All the ejaculates evaluated showed excellent semen quality characteristics; the sperm they contained was more rapid than horse sperm. Principal components analysis showed sperm l-lactate production to be a good predictor of semen condition. This, plus the characteristics of the motility patterns of the different sperm subpopulations, provides an excellent overall indicator of semen quality.

Animals↗

The effect of removal of seminal plasma, egg yolk level and season on sperm freezability of canary buck (Capra hircus).

Goat semen is different from that of other domestic species in its limited tolerance to the inclusion of egg yolk in the freezing medium, and this tolerance depends on the presence of enzymes in the seminal plasma that react with egg yolk, producing toxic compounds to the spermatozoa. Moreover, the goat is a seasonal breeder that shows variations in semen quality throughout the year, and those variations may affect semen freezability; hence in freezing protocols, for instance, removal of seminal plasma (washing) yields varying results. This work was designed to study this problem in Canary goats: semen from six males was collected in spring, autumn or winter, washed or non-washed, diluted in a freezing extender with 1.5, 6 or 12% egg yolk, frozen, and thawed after 2 days, 2 or 6 months of cryopreservation. The effect of egg yolk concentration in the freezing extender was far more important than the effect of washing or season on sperm cryosurvival. The quality of frozen-thawed semen tended to improve as egg yolk concentration increased regardless of the effects of season, washing or period of cryopreservation. Washing produced a positive effect on frozen-thawed semen collected during spring or autumn, but the difference decreased as the concentration of yolk increased. However, washing produced a negative effect on frozen-thawed semen collected during winter, diluted with either 6 or 12% egg yolk. There was no apparent seasonal effect on gross measures of sperm production but the seasonal effect was ever present and was reinforced by freezing.

Acrosome↗

Variations in the proportion of glycolytic/non-glycolytic energy substrates modulate sperm membrane integrity and function in diluted boar samples stored at 15-17 degrees C.

In this work the role of energy substrates in the maintenance of boar-sperm survival during storage at 15-17 degrees C was tested. For this purpose, boar spermatozoa were stored at 15-17 degrees C in several defined media with separate combinations of a monosaccharide, glucose and a non-monosaccharide, either citrate or lactate, energy substrates. Our results indicate that the medium containing the highest concentration of glucose together with low lactate levels was the most suitable to maintain sperm quality for 168 h at 15-17 degrees C. This was confirmed after observation of the results of the percentages of viability and altered acrosomes, the osmotic resistance test, the hyperosmotic resistance test and the rhythm of L-lactate production. The survival ability of boar sperm was greater in this experimental medium than in the standard Beltsville Thawing Solution extender, which contains only glucose as an energy substrate, although at a concentration far higher than that of all the tested experimental media. Our results indicate that the exact composition, more than the pure quantity of energy substrates, is a very important modulatory factor which affects survival ability of boar sperm in refrigeration. Thus, the exact combination of several energy substrates would have to be taken into account when optimizing the design of commercial extenders to store boar spermatozoa at 15-17 degrees C.

Acrosome↗

Semen characteristics and plasma levels of testosterone after bilateral vasectomy in bucks.

The effects of bilateral vasectomy on the seminal characteristics were assessed in six bucks of the Canarian breed. In addition, we tried to establish the effects of vasectomy on the plasmatic concentrations of testosterone and the libido of the bucks. Semen samples were collected once a week from 8 weeks before to 16 weeks after vasectomy; blood samples were collected prior to vasectomy, and then at once and 1 week after vasectomy and every 2 weeks from the week 4 to the end of the experiment. One week after the vasectomy, ejaculated spermatozoa were non-motile and the percentage of live spermatozoa was below 5% in all vasectomized males; in addition, the total number of cells/ejaculate was 3100 x 106 and 30 x 106 spermatozoa in the control and vasectomized males, respectively. Our results suggest that the vasectomized males may be used as oestrus detectors, without risks of accidental fecundating, only 1 week after vasectomy. Before vasectomy, no significant differences were observed in plasma levels of testosterone between the vasectomized and control males (5.4 +/- 1.2 and 3.9 +/- 1.4 ng/ml, respectively); from 4 to 12 weeks after vasectomy, a marked decrease in the testosterone concentration in all males (vasectomized and control bucks) was observed. From 12 weeks after vasectomy until the end of the experiment, four of the vasectomized males and the control males recovered their normal libido. The results suggest that vasectomy did not exert a remarkable effect on the steroidogenic functionality of the testicle.

Animals↗

Importance of cooling rate and animal variability for boar sperm cryopreservation: insights from the cryomicroscope.

A series of experiments was set up to investigate the effect of different cooling rates on boar sperm cryosurvival using cryomicroscopy. The cooling protocols were split into two stages: (i) from +5 degrees C to -5 degrees C and (ii) from -5 degrees C to -50 degrees C. Fluorescent probes (SYBR14 and propidium iodide) were used to monitor plasma membrane integrity during the entire process. Cooling rates in the range 3 degrees C min(-1) to 12 degrees C min(-1) did not cause significant damage to the sperm plasma membrane between +5 degrees C and -5 degrees C; however, spermatozoa cooled at 24 degrees C min(-1) to -5 degrees C were slightly damaged. Motility was not particularly sensitive to variations in cooling rate. Cooling rates in the range 15 degrees C min(-1) to 60 degrees C min(-1) did not produce differences in sperm cryosurvival during freezing between -5 degrees C and -50 degrees C, or after thawing. In addition, cooling rates in the range 3 degrees C min(-1) to 80 degrees C min(-1) did not produce significant differences in sperm cryosurvival. However, slow freezing (3 degrees C min(-1)) induced a slight increase in the percentage of plasma membrane-damaged spermatozoa (propidium iodide-positive) at -50 degrees C. Inter-ejaculate and inter-boar differences in sperm cryosurvival were manifested independently of cooling rate. The sperm plasma membrane remained intact (SYBR14-positive) during cooling and freezing, but upon rewarming, the plasma membrane of a high proportion of spermatozoa was damaged (propidium iodide-positive), indicating that rewarming is a critical step of the freezing-thawing process.

Analysis of Variance↗

GABAergic drugs inhibit amphetamine-induced distractibility in the rat.

Drugs facilitating GABAergic neurotransmission have been reported to block some behavioral actions of dopaminergic stimulation but not others. The present experiments were performed with the purpose to extend the range of behaviors in which the interaction between GABA and dopamine have been studied. The ability of the GABAB agonist baclofen and the GABA transaminase inhibitor sodium valproate to block the enhanced distractibility produced by amphetamine was evaluated in a procedure especially designed for analyzing drugs' effects on distractibility. Briefly, rats were trained to traverse a straight runway with a sucrose solution as reinforcement. Once the response had been acquired, an additional runway ending in an empty box was connected. The time spent investigating this additional runway is the measure of distractibility. Male rats treated with amphetamine, 1 mg/kg, displayed an increase of the time spent in the additional runway. Baclofen, 2.5 and 5 mg/kg, and sodium valproate, 100 and 200 mg/kg, had no effect on distraction behavior when administered alone. However, when these drugs were administered together with amphetamine, 1 mg/kg, they completely inhibited the effects of the stimulant on distractibility. These data show that distractibility is similar to discrimination learning with regard to the capacity of GABAergic drugs to block the effects of dopaminergic stimulation. It is different from locomotor activity, however, where GABAergic drugs are ineffective in this respect.

4-Aminobutyrate Transaminase↗

Cloning and sequencing of the gene encoding the outer-membrane protein Tbp1 from Actinobacillus pleuropneumoniae. Expression of Tbp1 and Tbp2.

Some Gram-negative bacteria possess a system for the uptake of iron from host's transferrin consisting of two outer-membrane proteins, namely transferrin-binding proteins 1 and 2. Here we present the cloning and sequencing of the gene encoding the transferrin-binding protein 1 from a serotype 1 isolate of the Gram-negative pathogen Actinobacillus pleuropneumoniae. We have also cloned for expression the Tbp1 and Tbp2 encoding sequences. Both proteins were produced in Escherichia coli and further purified. We also report the deduced Tbp1 amino acid sequence comparison to Tbp1s from related species.

Actinobacillus pleuropneumoniae↗

Assessment of boar sperm function in relation to freezing and storage.

The functions necessary for normal fertilization to occur in vivo or in vitro are examined and a rational approach to identifying the main features of a fertilizing spermatozoon are developed. It is concluded that methods for testing the quality of spermatozoa must probe the dynamic changes experienced by the spermatozoa during capacitation or under stressful incubation conditions. Recent developments in the multivariate analysis of sperm motility data are used to illustrate the success that can be achieved by this approach. Ideally, changes in sperm motility characteristics should be correlated with an assessment of capacitation status. However, until the capacitation status of any individual cell can be clearly defined this will remain problematic.

Animals↗

Cloning, sequencing and expression of the transferrin-binding protein 1 gene from Actinobacillus pleuropneumoniae.

Two outer-membrane proteins are involved in the uptake of iron from transferrin by certain Gram-negative bacteria, transferrin-binding proteins 1 and 2. The gene encoding transferrin-binding protein 1 from a serotype 1 isolate of the Gram-negative pathogen Actinobacillus pleuropneumoniae was cloned, and a fragment encoding 700 amino acids of Tbp1 was expressed in Escherichia coli. We also report here sequencing of the tbpl gene and a comparison of the deduced amino acid sequence with Tbpls from related species. The predicted polypeptide product of tbpl is a 106 kDa protein with a 22-residue signal peptide.

Actinobacillus pleuropneumoniae↗

Effects of frozen and other storage conditions on alpha-tocopherol content of cow milk.

The effect of temperature and length of storage on the stability of alpha-tocopherol in UHT milk was studied by HPLC. The effect of water activity conditions during storage of milk powder on the stability of the same vitamin was also studied. Losses of alpha-tocopherol occurred in UHT milk after 1 mo of storage at 30 degrees C; losses increased as storage time increased. In general, when the storage temperature changed from 30 to 40 degrees C, decrease in alpha-tocopherol content was greater. Short periods of frozen storage of the UHT milk (up to 60 d) had no effect on alpha-tocopherol content. However, frozen storage from 4 to 8 mo produced losses in alpha-tocopherol. When water activity was increased, the alpha-tocopherol content in milk powder decreased significantly after 2 mo of storage at 20 degrees C.

Animals↗

Stability of retinol in milk during frozen and other storage conditions.

The effect of storage temperature and storage time on the retinol content of four commercial unfortified whole UHT milk samples was studied by HPLC. Significant losses of retinol in these milks were observed after 1 month of storage at 30 degrees C. Losses generally increased with storage time. Increasing the storage temperature from 30 degrees C to 40 degrees C had a variable effect depending on the particular milk. Short periods of frozen storage (up to 60 days) had no effect on the retinol content. However, frozen storage time from 4 to 8 months led to significant (P < 0.05) losses in retinol content. The effect of water activity and temperature conditions during storage of unfortified whole milk powder on the stability of retinol was also studied. Increasing the activity of water and temperature of storage significantly lowered the retinol content in milk powder.

Animals↗

Is sperm cryopreservation at -150 degree C a feasible alternative?

A series of experiments was carried out to validate a -150 degree C ultra-low temperature freezer for its possible use to properly freeze and store semen. In the first part, crude sample handling was simulated to see whether temperature of stored samples was maintained within a safe range; also, the freezing point and latent heat of fusion plateau of a semen extender were monitored. In the second part, buck semen was (i) frozen in liquid nitrogen and stored in the ultra-low freezer, (ii) frozen and stored in the ultra-low freezer, and (iii) frozen and stored in liquid nitrogen, to compare sperm cryosurvival between freezing methods. Both, frequent removal of samples and long opening of the freezer door did not negatively affect stored sample temperature; latent heat of fusion plateau was 5 minutes long. Semen stored either at -150 degree C or at -196 degree C cryosurvived similarly after 2 days and after 2 months of cryopreservation.

Animals↗

A custom-built controlled-rate freezer for small sample cryopreservation studies.

A custom-built controlled-rate freezing machine is described. It operates by automatically raising and lowering the sample carrier in a static column of liquid nitrogen vapour. It is controlled by a computer-assisted thermocouple feedback system that operates a stepper motor driving the sample carrier. The cooling protocol is divided into three phases: cooling from +5 to -5 C, initiation of ice nucleation, and cooling from -5 to -80 C. Experiments are described to validate the device over a range of different cooling rates. A freezing protocol is established to cool samples in plastic straws over a range of rates up to 80 C/min, with rapid and consistent absorption of the latent heat.

Animals↗

[Electrocardiogram in interstitial pneumonia. Electrocardiographic-hemodynamic correlation].

Twenty four patients with interstitial pulmonary disease from the Cardiopulmonary Service of the Instituto Nacional de Cardiología were studied. Surface electrocardiograms and pulmonary hemodynamic studies were registered to all patients in order to obtain the pulmonary vascular resistances and pressures to find out the electrocardiographic-hemodynamic correlation. Three groups were made according to the pulmonary artery sistolic pressure: Group I, 30 to 45 mm Hg (6 patients); Group II, 46 to 70 mm Hg (12 patients) and Group III more of 60 mm Hg (6 patients). Also, arterial gaseous partial pressures were determined, among other pulmonary test parameters. Electrocardiographically, the rhythm, A=V node conduction, AQRSF, APF, atrial and ventricular hypertrophy, intraventricular conduction and ventricular repolarization were studied in leads II, III, aVF, aVR, V1, V2, V3 and V6. The characteristics of the ventricular repolarization and depolarization processes of each group are described, in order to discuss the mechanism of the anormalities. Finally, the electrocardiographic findings were correlated with the different degrees of pulmonary arterial hypertension; the conclusions are: I. The electrocardiogram allows us to differenciate the patients with III degree of arterial pulmonary hypertension, from those of I and II degrees. II. Patients with I and II degrees of pulmonary arterial hypertension determine nearly the same electrocardiographic alterations; the mean differences to establish the differential diagnosis is the AQRSF, shifted to the right and upwards, in the II degree of pulmonary arterial hypertension.

Adult↗