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1.
Biol Reprod ; 108(2): 324-337, 2023 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-36468681

RESUMO

Although recent research has addressed the impact of cryopreservation on the stallion sperm proteome, studies addressing the stallion sperm phosphoproteome are lacking. In the present study, the data set of proteomes of fresh and cryopreserved spermatozoa were reanalyzed, showing that cryopreservation caused significant changes in the phosphoproteome. The phosphoproteins reduced most significantly by cryopreservation were Ca2+binding tyrosine phosphorylation regulated, protein kinase cAMP-activated catalytic subunit beta (CABYR), mitochondria eating protein (SPATA18), A kinase anchoring protein 4 (AKAP4), A-kinase anchoring protein 3 (AKAP3) and the Family with sequence similarity 71 member B (FAM71B). These proteins belong to the gene ontology (GO) terms sperm fibrous sheath (GO: 0035686), and sperm principal piece (GO: 0097228). The regulatory interactions between kinases and phosphorylation sites on the proteins that were affected most were also investigated, and the potential kinases (based on human orthologs) involved in the regulation of these phosphoproteins identified were: PKCß for SPATA18 and GSK3ß for CABYR. Kinase inhibition assays were also conducted showing that kinases phosphorylating the above-mentioned proteins play an important role in their activity and thus, phosphorylation controls the activity of these proteins and their role in the regulation of the functionality and viability of stallion spermatozoa. In conclusion, the data reported here contribute to the understanding of the fact that the dephosphorylation of certain proteins is a molecular lesion induced by cryopreservation in the stallion spermatozoa.


Assuntos
Preservação do Sêmen , Sêmen , Masculino , Animais , Cavalos , Humanos , Sêmen/metabolismo , Espermatozoides/metabolismo , Cauda do Espermatozoide/metabolismo , Fosforilação , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Criopreservação/veterinária , Preservação do Sêmen/veterinária , Motilidade dos Espermatozoides/fisiologia , Proteínas de Ancoragem à Quinase A
2.
J Proteome Res ; 20(5): 2435-2446, 2021 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-33656888

RESUMO

Although cryopreservation is widely used in animal breeding, the technique is still suboptimal. The population of spermatozoa surviving the procedure experiences changes attributed to alteration in their redox regulation. In order to expand our knowledge regarding this particular aspect, the proteome in fresh and frozen thawed aliquots of equine spermatozoa was studied to identify the proteins most severely affected by the procedure. If alteration of redox regulation is a major factor explaining cryodamage, proteins participating in redox regulation should be principally affected. Using a split sample design, 30 ejaculates from 10 different stallions were analyzed as fresh spermatozoa, and another aliquot from the same ejaculate was analyzed as a frozen thawed sample. The proteome was studied under both conditions using UHPLC-MS/MS and bioinformatic analysis conducted to identify discriminant variables between both conditions. Data are available through the ProteomeXchange Consortium with identifier PXD022236. The proteins most significantly reduced were Aldo-keto reductase family 1 member B (p = 2.2 × 10-17) and Superoxide dismutase (Cu-Zn) (p = 4.7 × 10-14). This is the first time that SOD1 has been identified as a discriminating variable using bioinformatic analysis, where it was one of the most highly significantly different proteins seen between fresh and frozen thawed semen. This finding strongly supports the theory that alteration in redox regulation and oxidative stress is a major factor involved in cryodamage and suggests that control of redox regulation should be a major target to improve current cryopreservation procedures.


Assuntos
Criopreservação , Espectrometria de Massas em Tandem , Aldeído Redutase , Animais , Cavalos , Masculino , Oxirredutases , Motilidade dos Espermatozoides , Espermatozoides , Superóxido Dismutase/genética , Superóxido Dismutase-1/genética
3.
Biol Reprod ; 104(5): 1097-1113, 2021 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-33438027

RESUMO

The identification of stallions and or ejaculates that will provide commercially acceptable quality post-thaw before cryopreservation is of great interest, avoiding wasting time and resources freezing ejaculates that will not achieve sufficient quality to be marketed. Our hypothesis was that after bioinformatic analysis, the study of the stallion sperm proteome can provide discriminant variables able to predict the post-thaw quality of the ejaculate. At least three ejaculates from 10 different stallions were frozen following a split sample design. Half of the ejaculate was analyzed as a fresh aliquot and the other half was frozen and then analyzed as a frozen-thawed aliquot. Computer-assisted sperm analysis and flow cytometry were used to analyze sperm quality. Detailed proteomic analysis was performed on fresh and frozen and thawed aliquots, and bioinformatic analysis was used to identify discriminant variables in fresh samples able to predict the outcome of cryopreservation. Those with a fold change > 3, a P = 8.2e-04, and a q = 0.074 (equivalent to False discovery rate (FDR)) were selected, and the following proteins were identified in fresh samples as discriminant variables of good motility post-thaw: F6YTG8, K9K273, A0A3Q2I7V9, F7CE45, F6YU15, and F6SKR3. Other discriminant variables were also identified as predictors of good mitochondrial membrane potential and viability post-thaw. We concluded that proteomic approaches are a powerful tool to improve current sperm biotechnologies.


Assuntos
Criopreservação/veterinária , Cavalos/fisiologia , Proteoma/química , Análise do Sêmen/veterinária , Preservação do Sêmen/veterinária , Espermatozoides/fisiologia , Animais , Masculino , Espermatozoides/química
4.
Biol Reprod ; 105(2): 519-532, 2021 08 03.
Artigo em Inglês | MEDLINE | ID: mdl-33864078

RESUMO

Energy metabolism in spermatozoa is complex and involves the metabolism of carbohydrate fatty acids and amino acids. The ATP produced in the electron transport chain in the mitochondria appears to be crucial for both sperm motility and maintaining viability, whereas glycolytic enzymes in the flagella may contribute to ATP production to sustain motility and velocity. Stallion spermatozoa seemingly use diverse metabolic strategies, and in this regard, a study of the metabolic proteome showed that Gene Ontology terms and Reactome pathways related to pyruvate metabolism and the Krebs cycle were predominant. Following this, the hypothesis that low glucose concentrations can provide sufficient support for motility and velocity, and thus glucose concentration can be significantly reduced in the medium, was tested. Aliquots of stallion semen in four different media were stored for 48 h at 18°C; a commercial extender containing 67 mM glucose was used as a control. Stallion spermatozoa stored in media with low glucose (1 mM) and high pyruvate (10 mM) (LG-HP) sustained better motility and velocities than those stored in the commercial extender formulated with very high glucose (61.7 ± 1.2% in INRA 96 vs 76.2 ± 1.0% in LG-HP media after 48 h of incubation at 18°C; P < 0.0001). Moreover, mitochondrial activity was superior in LG-HP extenders (24.1 ± 1.8% in INRA 96 vs 51.1 ± 0.7% in LG-HP of spermatozoa with active mitochondria after 48 h of storage at 18°C; P < 0.0001). Low glucose concentrations may permit more efficient sperm metabolism and redox regulation when substrates for an efficient tricarboxylic acid cycle are provided. The improvement seen using low glucose extenders is due to reductions in the levels of glyoxal and methylglyoxal, 2-oxoaldehydes formed during glycolysis; these compounds are potent electrophiles able to react with proteins, lipids, and DNA, causing sperm damage.


Assuntos
Aldeídos/metabolismo , Metabolismo Energético , Glucose/deficiência , Cavalos/fisiologia , Ácido Pirúvico/metabolismo , Preservação do Sêmen/instrumentação , Espermatozoides/fisiologia , Animais , Masculino , Mitocôndrias , Oxirredução , Motilidade dos Espermatozoides/fisiologia
5.
Biol Reprod ; 103(6): 1275-1288, 2020 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-32857155

RESUMO

Some stallions yield ejaculates that do not tolerate conservation by refrigeration prior to artificial insemination (AI), showing improvement after removal of most of the seminal plasma (SP) by centrifugation. In this study, the SP-proteome of 10 different stallions was defined through high-performance liquid chromatography with tandem mass spectrometry and bioinformatic analysis in relation to the ability of the ejaculates to maintain semen quality when cooled and stored at 5°C. Stallions were classified into three groups, depending on this ability: those maintaining good quality after direct extension in a commercial extender (good), stallions requiring removal of seminal plasma (RSP) to maintain seminal quality (good-RSP), and stallions, unable to maintain good semen quality even after RSP (poor). Pathway enrichment analysis of the proteins identified in whole equine SP using human orthologs was performed using g: profiler showing enriched Reactome and the Kyoto Encyclopedia of Genes and Genomes pathways related to hexose metabolism, vesicle mediated transport, post translational modification of proteins and immune response. Specific proteins overrepresented in stallions tolerating conservation by refrigeration included a peroxiredoxin-6 like protein, and transcobalamin-2, a primary vitamin B12-binding, and transport protein. Also, the protein involved in protein glycosylation, ST3 beta-galactoside alpha-2,3-sialyltransferase 1 was present in good stallions. These proteins were nearly absent in poor stallions. Particularly, annexinA2 appeared as to be the most powerful discriminant variable for identification of stallions needing RSP prior to refrigeration, with a P = 0.002 and a q value = 0.005. Overall this is the first detailed study of the equine SP-proteome, showing the potential value of specific proteins as discriminant bio-markers for clinical classification of stallions for AI.


Assuntos
Anexinas/metabolismo , Cavalos/fisiologia , Refrigeração , Preservação do Sêmen/veterinária , Espermatozoides/fisiologia , Animais , Biomarcadores/química , Sobrevivência Celular/fisiologia , Masculino , Proteínas de Plasma Seminal/química , Proteínas de Plasma Seminal/metabolismo
6.
Biol Reprod ; 101(1): 208-222, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30998234

RESUMO

Oxidative stress is considered a major mechanism causing sperm damage during cryopreservation and storage, and underlies male factor infertility. Currently, oxidative stress is no longer believed to be caused only by the overproduction of reactive oxygen species, but rather by the deregulation of redox signaling and control mechanisms. With this concept in mind, here, we describe for the first time the presence of the soluble carrier family 7 member 11 (SLC7A11) antiporter, which exchanges extracellular cystine (Cyss) for intracellular glutamate, in stallion spermatozoa, as well as its impact on sperm function using the specific inhibitor sulfasalazine. Spermatozoa incubated with Cyss exhibited an increased intracellular GSH content compared with controls (P < 0.01): 50% in fresh extended stallion spermatozoa and 30% in frozen-thawed spermatozoa. This effect was prevented by the addition of sulfasalazine to the media. Cystine supplementation also reduced the oxidation-reduction potential of spermatozoa, with sulfasalazine only preventing this effect on fresh spermatozoa that were incubated for 3 h at 37°C, but not in frozen-thawed spermatozoa. While sulfasalazine reduced the motility of frozen-thawed spermatozoa, it increased motility in fresh samples. The present findings provide new and relevant data on the mechanism regulating the redox status of spermatozoa and suggest that a different redox regulatory mechanism exists in cryopreserved spermatozoa, thus providing new clues to improve current cryopreservation technologies and treat male factor infertility.


Assuntos
Sistema y+ de Transporte de Aminoácidos/metabolismo , Cistina/metabolismo , Cavalos/metabolismo , Espermatozoides/metabolismo , Animais , Cistationina gama-Liase/metabolismo , Cistina/farmacologia , Glutationa/metabolismo , Masculino , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Análise do Sêmen/métodos , Análise do Sêmen/veterinária , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Espermatozoides/efeitos dos fármacos
7.
Reprod Fertil Dev ; 30(11): 1462-1471, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29843893

RESUMO

Herein we describe a new protocol to induce boar sperm hypermotility: temperature-induced hypermotility (TIH). Briefly, spermatozoa stored at 17°C in a calcium-free Tyrode's basal medium (containing EGTA) were exposed to increased temperature by incubation at 38.5°C. Hypermotility induced by the calcium ionophore A23187 was used as a control (calcium ionophore-induced hyperactivity (CIIH)). The increase in temperature led to an increase in the percentage of hypermotile spermatozoa. When the slope of the temperature increase is near zero, sperm hyperactivity becomes a more progressive movement. Motility parameters of sperm hyperactivation induced by TIH were different from those following CIIH. Cluster analysis revealed that these two populations of hyperactivated spermatozoa are different. TIH is independent of extracellular Ca2+ but dependent on intracellular Ca2+ release. Moreover, TIH is unaffected by protein kinase A (PKA) inhibition, whereas CIIH is reduced by half in the presence of a PKA inhibitor. In conclusion, we have demonstrated that: (1) a temperature increase in boar spermatozoa is a stimulus that can induce a hyperactive population, which is differs from the hyperactive sperm population induced by calcium ionophore; (2) the temperature increase in spermatozoa triggers the release of Ca2+ from intracellular stores; (3) extracellular calcium is not required for TIH; and (4) TIH in boar spermatozoa is independent of PKA activity.


Assuntos
Cálcio/metabolismo , Transdução de Sinais/fisiologia , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/fisiologia , Temperatura , Animais , Calcimicina/farmacologia , Ionóforos de Cálcio/farmacologia , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Masculino , Inibidores de Proteínas Quinases/farmacologia , Transdução de Sinais/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Suínos
8.
Reprod Domest Anim ; 53 Suppl 2: 37-45, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30238662

RESUMO

Flow cytometry is increasingly used in research and also in clinical andrology. Recent developments in instrumentation, availability of probes and bioinformatics expand the possibilities of flow cytometry well beyond the classical two parametric analyses in use. In this paper, an overview of recent developments in flow cytometry will be presented under the perspective of the authors; aspects such a multicolor assays and computational cytometry will be discussed as well.


Assuntos
DNA/análise , Citometria de Fluxo , Mitocôndrias/fisiologia , Espermatozoides/fisiologia , Animais , Membrana Celular/fisiologia , Corantes , Masculino , Mitocôndrias/ultraestrutura , Espermatozoides/ultraestrutura
9.
Nutrients ; 15(3)2023 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-36771202

RESUMO

Physical training produces changes in the concentrations of trace mineral elements. Sex differences in copper (Cu) concentrations in athletes are scarce. The objectives of this study were (i) to analyze changes in intracellular (erythrocytes and platelets) and extracellular (plasma and urine) Cu concentrations during a sports season in soccer players and (ii) to analyze sex differences. A total of 46 soccer players (22 men and 24 women) participated in the study. Three assessments were performed throughout the sports season. Anthropometry, body composition, nutritional intake, physical condition, female hormones (menstrual cycle) and hematology were evaluated, as well as Cu determination (plasma, urine, erythrocytes, and platelets). Regarding longitudinal differences, there were discrepancies in plasma, urine, absolute erythrocyte, and absolute platelet Cu concentrations (p < 0.05). There were differences between sexes in Cu concentrations in urine, erythrocytes relative to cell number and in platelets relative to cell number (p < 0.05). During a sports season, there are changes in Cu concentrations in soccer players. Likewise, there could be sex differences in urinary, erythrocyte and platelet Cu concentrations.


Assuntos
Futebol , Humanos , Masculino , Feminino , Cobre , Caracteres Sexuais , Estações do Ano , Composição Corporal
10.
Nutrients ; 15(8)2023 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-37111051

RESUMO

Iron (Fe) metabolism and concentrations change during a sports season. Fe deficiency affects a significant number of women athletes. The aims of the present study were: (i) to analyze changes in hematological parameters of Fe status and (ii) to analyze changes in Fe concentrations in different biological matrices (serum, plasma, urine, erythrocytes, and platelets) during a sports season. Twenty-four Spanish semi-professional women's soccer players (23.37 ± 3.95 years) participated in the present study. Three assessments were performed throughout the sports season (beginning, middle and end of the season). Nutritional intake was evaluated and female hormones, hematological parameters of Fe status and Fe concentrations in plasma, serum, urine, erythrocytes and platelets were determined. There were no differences in Fe intake. Hemoglobin and mean corpuscular hemoglobin concentrations increased at the end of the season compared to initial values (p < 0.05). There were no significant changes in extracellular Fe concentrations (plasma, serum, and urine). However, erythrocyte Fe concentrations were lower at the end of the season (p < 0.05). Hematological parameters of Fe status and intracellular Fe concentrations change throughout the sports season in women's soccer players.


Assuntos
Ferro , Futebol , Humanos , Feminino , Estações do Ano , Atletas , Ingestão de Alimentos
11.
Environ Toxicol Pharmacol ; 99: 104107, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36914057

RESUMO

PURPOSE: The present study aimed to analyse sex differences in cadmium and lead concentrations in plasma, urine, platelets and erythrocytes and to relate these concentrations to biomarkers of iron status. METHODS: A total of 138 soccer players divided according to sex: men (n = 68) and women (n = 70) participated in the present study. All participants resided in the city of Cáceres (Spain). Erythrocyte, haemoglobin, platelet, plateletcrit, ferritin and serum iron values were determined. Cadmium and lead concentrations were quantified by inductively coupled plasma mass spectrometry. RESULTS: The women had lower haemoglobin, erythrocyte, ferritin and serum iron values (p < 0.01). Regarding cadmium, the women showed higher concentrations in plasma, erythrocytes and platelets (p < 0.05). As for lead, they also showed higher concentrations in plasma, relative values of erythrocytes and relative values of platelets (p < 0.05). Significant correlations were observed between cadmium and lead concentrations with biomarkers of iron status. CONCLUSIONS: Cadmium and lead concentrations are different between sexes. Biological differences between sexes and iron status could influence cadmium and lead concentrations. Lower serum iron concentrations and markers of Fe status increase Cd and Pb concentrations. Ferritin and serum iron have been directly related to increased Cd and Pb excretion.


Assuntos
Cádmio , Ferro , Humanos , Feminino , Masculino , Cádmio/análise , Chumbo/análise , Caracteres Sexuais , Ferritinas , Hemoglobinas , Atletas , Biomarcadores
12.
Toxics ; 11(11)2023 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-37999572

RESUMO

Aluminum (Al) is one of the most abundant trace mineral elements in the earth's crust. Al is considered a potent neurotoxicant. Physical exercise could cause modifications in some trace mineral elements. On the other hand, there could be sex differences in the exposure and deposits of toxic mineral elements. The aim of the present study was to compare sex and seasonal differences in extracellular and intracellular Al concentrations in football players. The study involved 22 male and 24 female football players from the fifth and second national category, respectively. Three assessments were carried out during the season (beginning, middle and end). Al concentrations in plasma, urine, erythrocytes and platelets were determined. Male football players ingested more Al (p < 0.05). Higher plasma Al concentrations were reported in male football players (p < 0.01). On the other hand, in both groups, increases and decreases in Al in the plasma and urine were observed in the second and third assessment, respectively (p < 0.01). There were sex differences in platelet Al concentrations (p < 0.05). Plasma and platelet Al concentrations may be different between the sexes. Al concentrations may change over the course of a season in football players.

13.
Reproduction ; 144(1): 67-75, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22573827

RESUMO

During the capacitation process, spermatozoa acquire the ability to fertilize an oocyte, and upregulation of cAMP-dependent protein tyrosine phosphorylation occurs. Recently, Src family tyrosine kinase (SFK) has been involved in spermatozoa capacitation as a key PKA-dependent tyrosine kinase in several species. This work investigates the expression and role of SFK in porcine spermatozoa. SFK members Lyn and Yes are identified in porcine spermatozoa by western blotting as well as two proteins named SFK1 and SFK2 were also detected by their tyrosine 416 phosphorylation, a key residue for SFK activation. Spermatozoa with SFK1 and SFK2 increase their Y416 phosphorylation time-dependently under capacitating conditions compared with noncapacitating conditions. The specific SFK inhibitor SU6656 unaffected porcine spermatozoa motility or viability. Moreover, SFK inhibition in spermatozoa under capacitating conditions leads to a twofold increase in both nonstimulated and calcium-induced acrosome reaction. Our data show that capacitating conditions lead to a time-dependent increase in actin polymerization in boar spermatozoa and that long-term incubation with SFK inhibitor causes a reduction in the F-actin content. In summary, this work shows that the SFK members Lyn and Yes are expressed in porcine spermatozoa and that SFK1 and SFK2 are phosphorylated (activated) during capacitation. Our results point out the important role exerted by SFK in the acrosome reaction, likely mediated in part by its involvement in the actin polymerization process that accompanies capacitation, and rule out its involvement in porcine spermatozoa motility.


Assuntos
Reação Acrossômica/fisiologia , Motilidade dos Espermatozoides/fisiologia , Suínos , Quinases da Família src/fisiologia , Reação Acrossômica/efeitos dos fármacos , Actinas/análise , Animais , Cálcio/farmacologia , Ativação Enzimática , Indóis/farmacologia , Masculino , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-yes/análise , Capacitação Espermática/fisiologia , Espermatozoides/enzimologia , Sulfonamidas/farmacologia , Quinases da Família src/análise , Quinases da Família src/antagonistas & inibidores
14.
Antioxid Redox Signal ; 37(7-9): 521-537, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35180830

RESUMO

Significance: Proper functionality of the spermatozoa depends on the tight regulation of their redox status; at the same time these cells are highly energy demanding and in the energetic metabolism, principally in the electron transport chain in the mitochondria, reactive oxygen species are continuously produced, in addition to that observed in the Krebs cycle and during the ß-oxidation of fatty acids. Recent Advances: In addition, in glycolysis, elimination of phosphate groups from glyceraldehyde 3-phosphate and dihydroxyacetone phosphate results in the byproducts glyoxal (G) and methylglyoxal (MG); these products are 2-oxoaldehydes. The presence of adjacent carbonyl groups makes them strong electrophiles that react with nucleophiles in proteins, lipids, and DNA, forming advanced glycation end products. Critical Issues: This mechanism is behind subfertility in diabetic patients; in the animal breeding industry, commercial extenders for stallion semen contain a supraphysiological concentration of glucose that promotes MG production, constituting a potential model of interest. Future Directions: Increasing our knowledge of sperm metabolism and its interactions with redox regulation may improve current sperm technologies in use, and shall provide new clues to understanding infertility in males. Moreover, stallion spermatozoa due to its accessibility, intense metabolism, and suitability for proteomics/metabolomic studies may constitute a suitable model for studying regulation of metabolism and interactions between metabolism and redox homeostasis. Antioxid. Redox Signal. 37, 521-537.


Assuntos
Sêmen , Motilidade dos Espermatozoides , Animais , Cavalos , Masculino , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Sêmen/metabolismo , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/metabolismo
15.
Anim Reprod Sci ; 246: 106805, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34275685

RESUMO

An overview of the sperm metabolism is presented; using the stallion as a model we review glycolysis, Krebs Cycle and oxidative phosphorylation, paying special attention to the interactions among them. In addition, metabolism implies a series of coordinated oxidation-reduction reactions and in the course of these reactions reactive oxygen species (ROS) and reactive oxoaldehydes are produced ; the electron transport chain (ETC) in the mitochondria is the main source of the anion superoxide and hydrogen peroxide, while glycolysis produces 2-oxoaldehydes such as methylglyoxal as byproducts; due to the adjacent carbonyl groups are strong electrophiles (steal electrons oxidizing other compounds). Sophisticated mechanisms exist to maintain redox homeostasis, because ROS under controlled production also have important regulatory functions in the spermatozoa. The interactions between metabolism and production of reactive oxygen species are essential for proper sperm function, and deregulation of these processes rapidly leads to sperm malfunction and finally death. Lastly, we briefly describe two techniques that will expand our knowledge on sperm metabolism in the coming decades, metabolic flow cytometry and the use of the "omics" technologies, proteomics and metabolomics, specifically the micro and nano proteomics/metabolomics. A better understanding of the metabolism of the spermatozoa will lead to big improvements in sperm technologies and the diagnosis and treatment of male factor infertility.


Assuntos
Doenças dos Cavalos , Infertilidade Masculina , Masculino , Cavalos , Animais , Motilidade dos Espermatozoides/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Ciclo do Ácido Cítrico , Sêmen/metabolismo , Espermatozoides/fisiologia , Infertilidade Masculina/veterinária , Estresse Oxidativo , Doenças dos Cavalos/metabolismo
16.
Artigo em Inglês | MEDLINE | ID: mdl-36554736

RESUMO

The objectives of this study were to analyze the injuries suffered during the previous year by amateur padel players according to the characteristics of the racket, their usual volume of practice and their experience in padel. A total of 950 amateur players (X age: 31.68 years; X weight: 70.84 kg; X height: 170.9 cm) participated voluntarily, completing an ad-hoc questionnaire. The results indicated that the appearance of the injuries and their location was different according to the sex of the amateur padel players. Men had a higher incidence of muscle and ligament injuries in the shoulder, and tendon injuries in the elbow. On the other hand, women had a greater probability of having muscle injuries in the shoulder and arm, ligament injuries in the elbow and bone injuries in the wrist and elbow. In general, tendon injuries were the most common injury in padel and the shoulder and elbow were the most affected areas. Moreover, men tend to use heavy (CSR = 6.0), fiberglass or carbon (CSR = 2.1), diamond-shaped rackets (CSR = 3.2), with a hard core (CSR = 4.4) and with two or more over grips (CSR = 2.7). Women usually use less heavy (CSR = 6.0), round-shaped rackets (CSR = 4.9), with a soft core (CSR = 4.4) and with one or no over grips (CSR = 2.7). In addition, men tend to play padel more often and have been practicing for longer. In conclusion, although the risk of injury depends on many factors, we identified that the characteristics of the racket, the volume of weekly practice, the experience of the player and the gender of the player are fundamental aspects to take into account for the prevention of injuries in amateur padel players.


Assuntos
Traumatismos em Atletas , Traumatismos dos Tendões , Masculino , Humanos , Feminino , Adulto , Incidência , Atletas , Ombro , Cotovelo , Traumatismos em Atletas/epidemiologia
17.
Theriogenology ; 177: 94-102, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34687941

RESUMO

Seminal plasma plays an important role in sperm physiology. Seminal plasma proteins vehiculated in microvesicles, carry RNAs and proteins with a potential role in early embryo development. Additionally, proteins present in seminal plasma participate in redox regulation and energy metabolism. In view of these facts, we hypothesized that differences in protein composition of the seminal plasma among stallions may help to explain differences in freeze-ability seen among them. Three independent ejaculates from 10 different stallions of varying breeds were frozen using standard protocols in our laboratory. Aliquots of the ejaculate were separated and stored at -80 °C until further proteomic analysis. Semen analysis was performed using computer assisted sperm analysis and flow cytometry. Significant differences in proteome composition of seminal plasma were observed in the group of stallions showing better motility post thaw. 3116 proteins were identified, and of these, 34 were differentially expressed in stallions with better motility post thaw, 4 of them were also differentially expressed in stallions with different percentages of linearly motile sperm post thaw and 1 protein, Midasin, was expressed in stallions showing high circular velocity post thaw. Seminal plasma proteins may play a major role in sperm functionality; being vehiculated through extracellular vesicles and participating in sperm physiology. Bioinformatic analysis identifies discriminant proteins able to predict the outcome of cryopreservation, identifying potential new biomarkers to assess ejaculate quality.


Assuntos
Preservação do Sêmen , Adenina , Animais , Arginina , Criopreservação/veterinária , Cavalos , Masculino , Metiltransferases , Proteômica , Sêmen , Preservação do Sêmen/veterinária , Proteínas de Plasma Seminal , Motilidade dos Espermatozoides , Espermatozoides
18.
Theriogenology ; 177: 34-41, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34656835

RESUMO

Seminal plasma proteins have important roles in sperm functionality, and different mechanisms including micro-vesicle transport of proteins are involved in the regulation of sperm biology. Due to the role of seminal plasma, we hypothesized that specific proteins present in seminal plasma may be used as discriminant variables with potential to identify stallions producing different quality ejaculates; 10 fertile stallions, with different motility and velocity values (although within normal ranges) were used in this study. Motilities and velocities were studied using computer assisted sperm analysis (CASA), while protein composition of the seminal plasma was studied using UHPLC-MS/MS. Specific proteins were more abundant in samples with poorer percentages of total motility, average path velocity and circular velocity, and were: Secreted phosphoprotein 1, Fructose-bisphosphate aldolase (p = 1,95E-09; q = 0.0005) and Malate dehydrogenase 1 (p = 1,41E-11; q = 0.002), to the contrary samples with better straight-line velocity values were enriched in Glutathione peroxidase (p=0.00013; q=0.04) and Triosephosphate isomerase (p=0.00015; q=0.04).


Assuntos
Proteínas de Plasma Seminal , Motilidade dos Espermatozoides , Animais , Biomarcadores , Cavalos , Masculino , Sêmen , Espermatozoides , Espectrometria de Massas em Tandem/veterinária
19.
Data Brief ; 45: 108578, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36131951

RESUMO

This paper provides a detailed set of data on how the stallion sperm proteome differs among stallions with different sperm motilities, although within normal ranges. Findings distinguish proteins that may help to identify stallions of superior sperm motility. Sperm proteins were analyzed using a UHPLC/MS/MS system comprising of an Agilent 1290 infinity series UHPLC coupled to an Agilent 6550 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA). These data can be used to disclose potential targets to identify good sperm samples and to study specific pathways involved in the regulation of sperm motility. This data article is linked to the paper "Proteins involved in mitochondrial metabolic functions and fertilization predominate in stallions with better motility Journal of Proteomics 247:104335 doi: 10.1016/j.jprot.2021.104335".

20.
Nutrients ; 14(9)2022 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-35565824

RESUMO

Trace mineral element concentrations are under homeostatic control. Selenium (Se) is a very important micronutrient for the antioxidant and immune system. Se metabolism could be modified due to physical training. This research aimed to analyze the extracellular (plasma, urine and serum) and intracellular (platelets and erythrocytes) concentrations of Se in athletes and to compare it with subjects with low levels of physical training. Forty young men divided into a control group (CG; n = 20; 19.25 ± 0.39 years) and a training group (TG; n = 20; 18.15 ± 0.27 years) participated in this study. The TG was formed by semi-professional soccer players. The analysis of Se was determined by inductively coupled plasma mass spectrometry. The TG obtained higher values of maximum oxygen consumption and muscle percentage (p < 0.05). The TG showed reduced absolute (p < 0.01) and relative (p < 0.05) Se concentrations in erythrocytes and platelets in comparison to CG. Trace element assessments should not be limited only to extracellular compartments as there could be deficiencies at the intracellular level.


Assuntos
Selênio , Oligoelementos , Atletas , Eritrócitos/química , Exercício Físico/fisiologia , Humanos , Masculino
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