ABSTRACT
RESUMEN Objetivo: Determinar los factores asociados con el agotamiento psicológico (AP) en odontólogos especialistas que laboraban en la ciudad de Bucaramanga y su área metropolitana. Métodos: Se realizó un estudio observacional analítico de corte transversal en el que se aplicó la versión validada del cuestionario Maslach Burnout Inventory-Human Services Survey (MBI-HSS) a 117 especialistas del área de odontología. Se analizaron los puntajes obtenidos en las 3 dimensiones del instrumento, las características sociodemográficas, los aspectos del entorno laboral y profesional y los hábitos. En el análisis bivariado, se utilizó el coeficiente de correlación de Spearman, la prueba de la U de Mann Whitney o Kruskal-Wallis y en el ánalisis multivariado, se realizó una regresión logística. Un valor de p<0,05 se consideró estadísticamente significativo. Cada participante aceptó su inclusión en el estudio tras el proceso de consentimiento informado. Resultados: El promedio de edad fue 44,0 ± 7,8 años y el 57,3% eran mujeres. El 3,4, el 4,3 y el 4,3% de los participantes tenían altos grados de agotamiento emocional, despersonaliza ción y falta de realización personal respectivamente. El consumo de cigarrillo presentó una asociación directa con las dimensiones despersonalización (p = 0,031) y falta de realización personal (p = 0,025). De otra parte, tener 10 años o más de egresado del posgrado evidenció una asociación inversa en estas 2 dimensiones (p = 0,049 y p = 0,045 respectivamente). Conclusiones: Los resultados indican que el AP no es un problema frecuente en los odontó logos especialistas que laboran en Bucaramanga y su área metropolitana; sin embargo, es importante tener en cuenta que se observó relación del AP con el hábito de fumar y los años de egresado de la especialidad.
ABSTRACT Objective: To establish the factors associated with burnout syndrome in dental specialists working in the city of Bucaramanga and its metropolitan area. Methods: A cross-sectional study was conducted in which the validated version of the Maslach Burnout Inventory-Human Services Survey (MBI-HSS) questionnaire was applied to 117 dental specialists. The variables analysed were the scores obtained in the three dimensions of the instrument - sociodemographic and job-related characteristics, professional environment, and habits. Spearman's correlation coefficient and the Mann Whitney or Kruskal-Wallis U test were used in the bivariate analysis, and a logistic regression was perfor med in the multivariate analysis. A value of p < 0.05 was considered statistically significant. Each participant accepted their inclusion in the study after the Informed Consent process was done. Results: The average age was 44.0 ± 7.8 years, and 57.3% were women. It was observed that 3.4%, 4.3% and 4.3% of respondents scored highly in emotional exhaustion, depersonalisation and lack of personal achievement, respectively. Cigarette smoking presented a direct association with the dimensions of depersonalisation (p = 0.031) and lack of personal achie vement (p = 0.025). On the other hand, having completed the postgraduate degree 10 years or more ago showed a negative association in these two dimensions (p = 0.049 and p = 0.045, respectively). Conclusions: The results suggest that burnout syndrome is not a frequent problem in dental specialists who work in Bucaramanga and its metropolitan area. However, it is important to keep in mind that a relationship was observed between the syndrome and smoking, and the years after graduating in the specialty.
ABSTRACT
OBJECTIVE: To establish the factors associated with burnout syndrome in dental specialists working in the city of Bucaramanga and its metropolitan area. METHODS: A cross-sectional study was conducted in which the validated version of the Maslach Burnout Inventory-Human Services Survey (MBI-HSS) questionnaire was applied to 117 dental specialists. The variables analysed were the scores obtained in the three dimensions of the instrument - sociodemographic and job-related characteristics, professional environment, and habits. Spearman's correlation coefficient and the Mann Whitney or Kruskal-Wallis U test were used in the bivariate analysis, and a logistic regression was performed in the multivariate analysis. A value of p < 0.05 was considered statistically significant. Each participant accepted their inclusion in the study after the Informed Consent process was done. RESULTS: The average age was 44.0 ± 7.8 years, and 57.3% were women. It was observed that 3.4%, 4.3% and 4.3% of respondents scored highly in emotional exhaustion, depersonalisation and lack of personal achievement, respectively. Cigarette smoking presented a direct association with the dimensions of depersonalisation (p = 0.031) and lack of personal achievement (p = 0.025). On the other hand, having completed the postgraduate degree 10 years or more ago showed a negative association in these two dimensions (p = 0.049 and p = 0.045, respectively). CONCLUSIONS: The results suggest that burnout syndrome is not a frequent problem in dental specialists who work in Bucaramanga and its metropolitan area. However, it is important to keep in mind that a relationship was observed between the syndrome and smoking, and the years after graduating in the specialty.
Subject(s)
Burnout, Professional , Job Satisfaction , Humans , Female , Adult , Middle Aged , Male , Cross-Sectional Studies , Burnout, Psychological , Burnout, Professional/epidemiology , Burnout, Professional/psychology , Surveys and QuestionnairesABSTRACT
BACKGROUND: ALK rearrangements are present in 5% of nonsmall cell lung cancer (NSCLC) tumors and identify patients who can benefit from ALK inhibitors. ALK fusions testing using liquid biopsies, although challenging, can expand the therapeutic options for ALK-positive NSCLC patients considerably. RNA inside extracellular vesicles (EVs) is protected from RNases and other environmental factors, constituting a promising source for noninvasive fusion transcript detection. METHODS: EVs from H3122 and H2228 cell lines, harboring EML4-ALK variant 1 (E13; A20) and variant 3 (E6a/b; A20), respectively, were successfully isolated by sequential centrifugation of cell culture supernatants. EVs were also isolated from plasma samples of 16 ALK-positive NSCLC patients collected before treatment initiation. RESULTS: Purified EVs from cell cultures were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and flow cytometry. Western blot and confocal microscopy confirmed the expression of EV-specific markers as well as the expression of EML4-ALK-fusion proteins in EV fractions from H3122 and H2228 cell lines. In addition, RNA from EV fractions derived from cell culture was analyzed by digital PCR (dPCR) and ALK-fusion transcripts were clearly detected. Similarly, plasma-derived EVs were characterized by NTA, flow cytometry, and the ExoView platform, the last showing that EV-specific markers captured EV populations containing ALK-fusion protein. Finally, ALK fusions were identified in 50% (8/16) of plasma EV-enriched fractions by dPCR, confirming the presence of fusion transcripts in EV fractions. CONCLUSIONS: ALK-fusion transcripts can be detected in EV-enriched fractions. These results set the stage for the development of EV-based noninvasive ALK testing.
Subject(s)
Carcinoma, Non-Small-Cell Lung , Extracellular Vesicles , Lung Neoplasms , Carcinoma, Non-Small-Cell Lung/pathology , Cell Line , Extracellular Vesicles/metabolism , Humans , Lung Neoplasms/pathology , Oncogene Proteins, Fusion/genetics , RNA , Receptor Protein-Tyrosine Kinases/geneticsABSTRACT
OBJECTIVE: To establish the factors associated with burnout syndrome in dental specialists working in the city of Bucaramanga and its metropolitan area. METHODS: A cross-sectional study was conducted in which the validated version of the Maslach Burnout Inventory-Human Services Survey (MBI-HSS) questionnaire was applied to 117 dental specialists. The variables analysed were the scores obtained in the three dimensions of the instrument - sociodemographic and job-related characteristics, professional environment, and habits. Spearman's correlation coefficient and the Mann Whitney or Kruskal-Wallis U test were used in the bivariate analysis, and a logistic regression was performed in the multivariate analysis. A value of p<0.05 was considered statistically significant. Each participant accepted their inclusion in the study after the Informed Consent process was done. RESULTS: The average age was 44.0±7.8 years, and 57.3% were women. It was observed that 3.4%, 4.3% and 4.3% of respondents scored highly in emotional exhaustion, depersonalisation and lack of personal achievement, respectively. Cigarette smoking presented a direct association with the dimensions of depersonalisation (p=0.031) and lack of personal achievement (p=0.025). On the other hand, having completed the postgraduate degree 10 years or more ago showed a negative association in these two dimensions (p=0.049 and p=0.045, respectively). CONCLUSIONS: The results suggest that burnout syndrome is not a frequent problem in dental specialists who work in Bucaramanga and its metropolitan area. However, it is important to keep in mind that a relationship was observed between the syndrome and smoking, and the years after graduating in the specialty.
ABSTRACT
The concentrations of cytokines in seminal plasma (SP) fluctuate over time in healthy males, weakening their practical usefulness as diagnostic tools. This study evaluated the relevance of intra-male variability in SP cytokines and to what extent the period of the year when ejaculate is collected contributes to such variability. Thirteen cytokines (GM-CSF, IFNγ, IL-1α, IL-1ß, IL-1ra, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18, and TNFα) were measured using the Luminex xMAP® technology for 180 SP samples of ejaculate collected over a year from nine healthy and fertile boars. The SP samples were grouped into two annual periods according to decreasing or increasing daylight and ambient temperature. Intra-male variability was higher than inter-male variability for all cytokines. All SP cytokines showed concentration differences between the two periods of the year, showing the highest concentration during the increasing daylength/temperature period, irrespective of the male. Similarly, some cytokines showed differences between daylength/temperature periods when focusing on their total amount in the ejaculate. No strong relationship (explaining more than 50% of the total variance) was found between annual fluctuations in SP-cytokine levels and semen parameters. In conclusion, the period of the year during which ejaculates were collected helps explain the intra-male variability of SP-cytokine levels in breeding boars.
ABSTRACT
Proteins in semen, either in spermatozoa (SPZ) or seminal plasma (SP), are directly involved in molecular processes and biological pathways regulating sperm function, including fertilizing ability. Therefore, semen proteins are candidates of choice for biomarkers discovery for fertility and for sperm (dys)function. Mass spectrometry (MS)-based proteomics has opened up a new era for characterizing and quantifying the protein profile of SP and SPZ, as well as for unveiling the complex protein interactions involved in the activation of sperm functionality. This article overviews existing literature on MS-based proteomics regarding porcine semen, with a specific focus on the potential practical application of the results achieved so far. The weaknesses of current studies and the perspectives for future research in MS-based pig semen proteomics are also addressed.
Subject(s)
Proteomics/methods , Semen Preservation/veterinary , Semen/metabolism , Seminal Plasma Proteins/metabolism , Swine , Animals , Male , Semen Analysis/veterinaryABSTRACT
BACKGROUND AND AIMS: One of the most distinctive features of desiccation-tolerant plants is their high cell wall (CW) flexibility. Most lichen microalgae can tolerate drastic dehydration-rehydration (D/R) conditions; however, their mechanisms of D/R tolerance are scarcely understood. We tested the hypothesis that D/R-tolerant microalgae would have flexible CWs due to species-specific CW ultrastructure and biochemical composition, which could be remodelled by exposure to cyclic D/R. METHODS: Two lichen microalgae, Trebouxia sp. TR9 (TR9, adapted to rapid D/R cycles) and Coccomyxa simplex (Csol, adapted to seasonal dry periods) were exposed to no or four cycles of desiccation [25-30 % RH (TR9) or 55-60 % RH (Csol)] and 16 h of rehydration (100 % RH). Low-temperature SEM, environmental SEM and freeze-substitution TEM were employed to visualize structural alterations induced by D/R. In addition, CWs were extracted and sequentially fractionated with hot water and KOH, and the gel permeation profile of polysaccharides was analysed in each fraction. The glycosyl composition and linkage of the main polysaccharides of each CW fraction were analysed by GC-MS. KEY RESULTS: All ultrastructural analyses consistently showed that desiccation caused progressive cell shrinkage and deformation in both microalgae, which could be rapidly reversed when water availability increased. Notably, the plasma membrane of TR9 and Csol remained in close contact with the deformed CW. Exposure to D/R strongly altered the size distribution of TR9 hot-water-soluble polysaccharides, composed mainly of a ß-3-linked rhamnogalactofuranan and Csol KOH-soluble ß-glucans. CONCLUSIONS: Cyclic D/R induces biochemical remodelling of the CW that could increase CW flexibility, allowing regulated shrinkage and expansion of D/R-tolerant microalgae.
Subject(s)
Lichens , Microalgae , Cell Wall , Desiccation , Fluid TherapyABSTRACT
Background: Boar seminal plasma is rich in cytokines, which could influence the capability of spermatozoa to tolerate preservation. Objectives: To evaluate the involvement of boar seminal plasma cytokines in the changes experienced by boar spermatozoa during their storage, either in liquid or frozen state. Materials and Methods: In two separated experiments, semen samples from healthy and fertile boars were split in two aliquots, one centrifuged twice (1,500 ×g for 10 min) to harvest seminal plasma, whereas the other was either commercially extended (3 × 107 sperm/mL) and liquid-stored at 17°C during 144 h (n = 28, Experiment 1) or frozen-thawed using a standard 0.5 mL protocol (n = 27, Experiment 2). Sixteen cytokines were quantified using Luminex xMAP®. Sperm attributes (CASA-evaluated total and progressive motility; flow cytometry-evaluated sperm viability, production of intracellular H2O2 and O 2 ⢠- and levels of lipid peroxidation in viable spermatozoa) were evaluated either at 0, 72, or 144 h of liquid storage (Experiment 1) or before freezing and at 30- and 150-min post-thawing (Experiment 2). Results: Multiple linear regression models, with Bayesian approach for variable selection, revealed that the anti-inflammatory TGF-ß2, TGF-ß3, IL-1Ra, and IL-4 and the pro-inflammatory IL-8 and IL-18, predicted changes in sperm motility for liquid-stored semen while the anti-inflammatory IFN-γ was included in the models predicting changes in all sperm attributes for cryopreserved semen. Conclusion: Specific boar seminal plasma cytokines would contribute to modulate the structural and metabolic changes shown by spermatozoa during preservation, either in liquid or frozen state.
ABSTRACT
Seminal extracellular vesicles (EVs) include exosomes (ø 40-120 nm) and microvesicles (MVs, ø 120-1000 nm), which would be involved in multiple functional reproductive roles. The study aimed to establish which EV subtypes are present in pig semen, using a high-resolution flow cytometer to explore differences in their tetraspanin expression profile. The EVs were isolated from 12 pig ejaculates using serial ultracentrifugation and characterized by dynamic light scattering and electron microscopy for size and morphology as well as for tetraspanin expression using flow cytometry with Carboxyfluorescein succinimidyl ester (CFSE) and antibodies against CD9, CD63 and CD81. Pig semen contained a heterogeneous EV-population regarding size and morphology. Flow cytometric analysis demonstrated that the proportion of EVs expressing CD63 and CD9 was higher in MVs (P < 0.001 and P < 0.05, respectively) than in exosomes, while the opposite was true for CD81; higher (P < 0.001) in exosomes than in MVs. In conclusion, (1) the new generation of flow cytometers are able to accurately identify EVs and to gate them in two size-different populations named exosomes and MVs. (2) Tetraspanins CD9, CD63 and CD81 are present in both seminal EVs, albeit with exosomes and MVs differing in expression profiles, suggesting dissimilar cargo and binding affinity.
Subject(s)
Extracellular Vesicles/chemistry , Semen/chemistry , Swine , Tetraspanins/analysis , Animals , Exosomes/chemistry , Exosomes/ultrastructure , Extracellular Vesicles/ultrastructure , Flow Cytometry , Male , Swine/metabolism , Tetraspanin 28/analysis , Tetraspanin 29/analysis , Tetraspanin 30/analysisABSTRACT
Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate to the loss of sperm function during cryopreservation remains unsolved. The present study aimed to clarify this issue evaluating differential changes in the proteome of fresh and frozen-thawed pig spermatozoa retrieved from the cauda epididymis and the ejaculate of the same boars, with clear differences in cryotolerance. Spermatozoa were collected from 10 healthy, sexually mature, and fertile boars, and cryopreserved using a standard 0.5 mL-straw protocol. Total and progressive motility, viability, and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT spermatozoa were analyzed using a LC-ESI-MS/MS-based Sequential Window Acquisition of All Theoretical Spectra approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein-protein networks highlighted a set of proteins quantitatively altered in ejaculated spermatozoa, directly involved in mitochondrial functionality which would explain why ejaculated spermatozoa deteriorate during cryopreservation.
Subject(s)
Cryopreservation/veterinary , Semen Preservation/veterinary , Spermatozoa/cytology , Swine , Animals , Cryopreservation/methods , Ejaculation , Epididymis/cytology , Epididymis/metabolism , Fertilization in Vitro , Male , Proteome/metabolism , Semen Preservation/methods , Sperm Motility , Spermatozoa/metabolism , Swine/physiologyABSTRACT
The preservation of sperm functional parameters and fertility post-cryopreservation largely varies in the porcine, a species with a fractionated ejaculate. Although intrinsic individual differences have primarily been linked to this variation, differences in protein abundance among frozen-thawed (FT)-spermatozoa are far more relevant. This study, performed in two experiments, looked for proteomic quantitative differences between FT-sperm samples differing in post-thaw viability, motility, apoptosis, membrane lipid peroxidation and nuclear DNA fragmentation. The spermatozoa were either derived from the sperm-rich ejaculate fraction (SRF) or the entire ejaculate (Experiment 1) or from the first 10 mL of the SRF, the remaining SRF and the post-SRF (Experiment 2). Quantitative sperm proteomic differences were analysed using a LC-ESI-MS/MS-based SWATH approach. In Experiment 1, FT-spermatozoa from the SRF showed better preservation parameters than those from the entire ejaculate, with 26 Sus scrofa proteins with functional sperm relevance showing relative quantitative differences (FC ≥ 1.5) between sperm sources. In Experiment 2, FT-spermatozoa from the first 10 mL of the SRF and the remaining SRF were qualitatively better than those from the post-SRF, and 187 proteins showed relative quantitative differences among the three ejaculate sources. The results indicate that quantitative proteome differences are linked to sperm cryosurvival.
Subject(s)
Cell Membrane/metabolism , Cryopreservation/veterinary , Gene Expression Regulation , Proteome/analysis , Semen Preservation/veterinary , Semen/metabolism , Spermatozoa/metabolism , Animals , Ejaculation , Male , Sperm Motility , SwineABSTRACT
Proteins are essential for sperm function, including their fertilizing capacity. Pig spermatozoa, emitted in well-defined ejaculate fractions, vary in their functionality, which could be related to different sperm protein composition. This study aimed (i) to update the porcine sperm proteome and (ii) to identify proteins differentially expressed in mature spermatozoa from cauda epididymis and those delivered in separate ejaculate fractions. Ejaculates from nine mature and fertile boars were manually collected in three separate portions: the first 10 ml of the sperm-rich ejaculate fraction (SRF), the rest of the SRF and the post-SRF. The contents of cauda epididymides of the boars were collected post-mortem by retrograde duct perfusion, generating four different semen sources for each boar. Following centrifugation, the resulting pellets of each semen source were initially pooled and later split to generate two technical replicates per source. The resulting eight sperm samples (two per semen source) were subjected to iTRAQ-based 2D-LC-MS/MS for protein identification and quantification. A total of 1,723 proteins were identified (974 of Sus scrofa taxonomy) and 1,602 of them were also quantified (960 of Sus scrofa taxonomy). After an ANOVA test, 32 Sus scrofa proteins showed quantitative differences (p < 0.01) among semen sources, which was particularly relevant for sperm functionality in the post-SRF. The present study showed that the proteome of boar spermatozoa is remodeled during ejaculation involving proteins clearly implicated in sperm function. The findings provide valuable groundwork for further studies focused on identifying protein biomarkers of sperm fertility.
Subject(s)
Ejaculation , Proteome/analysis , Spermatozoa/metabolism , Animals , Chromatography, Liquid , Gene Expression Regulation , Male , Sus scrofa , Swine , Tandem Mass SpectrometryABSTRACT
This study aimed to elucidate the effect of seminal plasma (SP) from post-SRF on boar sperm freezability and, in addition, to determine the relevance of sperm itself to sustain cryopreservation, regardless of the SP surrounding them. Twelve ejaculates from three boars were manually collected in fractions/portions, P1: the first 10â¯mL of the SRF, P2: the rest of the SRF and the post-SRF. Immediately, samples were centrifuged to separate spermatozoa from the surrounding SP. Spermatozoa from P1 and P2â¯were then incubated with its own SP or that from post-SRF, diluted in BTS (1:1, v/v) at 17⯰C overnight before being frozen in 0.5â¯mL straws using a standard protocol. Sperm motility (total and progressive) deteriorated (Pâ¯<â¯0.05) when P1- or P2-sperm when incubated overnight in SP from post-SRF, while sperm viability differed between P1 and P2 (Pâ¯<â¯0.05) regardless of the SP they were incubated in. Post-thaw sperm quality and functionality differed between P1 and P2, regardless of the SP used for overnight pre-freezing incubation. Post-thaw motility (Pâ¯<â¯0.05) and viability (Pâ¯<â¯0.01), as well as plasma membrane fluidity (Pâ¯<â¯0.05) or lipid peroxidation values (Pâ¯<â¯0.01) were best in P1 sperm compared to those of P2. The protein profile of sperm from P1 and P2, analyzed by 2D-PAGE, showed qualitative differences, which suggest that sperm rather than SP would explain differences in sperm freezability between ejaculate fractions/portions. Use of P1 fraction spermatozoa seems thus optimal for cryopreservation.
Subject(s)
Cryopreservation/veterinary , Freezing , Semen Preservation/veterinary , Semen/chemistry , Spermatozoa/physiology , Swine/physiology , Animals , Apoptosis , Cell Membrane , Cell Survival , Gene Expression Regulation , Lipid Peroxidation , Male , Membrane Proteins , Proteins/genetics , Proteins/metabolism , Proteoglycans , Protozoan Proteins , Reactive Oxygen Species , Semen Preservation/methods , Sperm MotilityABSTRACT
A complete characterization of the proteome of seminal plasma (SP) is an essential step to understand how SP influences sperm function and fertility after artificial insemination (AI). The purpose of this study was to identify which among characterized proteins in boar SP were differently expressed among AI boars with significantly different fertility outcomes. A total of 872 SP proteins, 390 of them belonging specifically to Sus Scrofa taxonomy, were identified (Experiment 1) by using a novel proteomic approach that combined size exclusion chromatography and solid-phase extraction as prefractionation steps prior to Nano LC-ESI-MS/MS analysis. The SP proteomes of 26 boars showing significant differences in farrowing rate (n = 13) and litter size (n = 13) after the AI of 10â¯526 sows were further analyzed (Experiment 2). A total of 679 SP proteins were then quantified by the SWATH approach, where the penalized linear regression LASSO revealed differentially expressed SP proteins for farrowing rate (FURIN, AKR1B1, UBA1, PIN1, SPAM1, BLMH, SMPDL3A, KRT17, KRT10, TTC23, and AGT) and litter size (PN-1, THBS1, DSC1, and CAT). This study extended our knowledge of the SP proteome and revealed some SP proteins as potential biomarkers of fertility in AI boars.
Subject(s)
Fertility/genetics , Litter Size/genetics , Proteome/genetics , Semen/physiology , Animals , Biomarkers/metabolism , Chromatography, Gel , Female , Gene Expression , Gene Expression Profiling , Gene Ontology , Insemination, Artificial , Male , Molecular Sequence Annotation , Proteome/metabolism , Semen/chemistry , Semen Analysis , Solid Phase Extraction/methods , Spectrometry, Mass, Electrospray Ionization , Sperm Motility/physiology , Spermatozoa/cytology , Spermatozoa/physiology , SwineABSTRACT
The paraoxonase type 1 (PON1) is an enzyme with antioxidant properties recently identified in the seminal plasma (SP) of several species, including the porcine. The aims of the present study were to (1) describe the immunohistochemical localisation of PON1 in the genital organs of fertile boars and (2) evaluate the relationship among PON1 activity and high-density lipoprotein cholesterol (HDL-C) concentration in fluids of the boar genital organs. Immunohistochemical analysis demonstrated that PON1 was present in testis (specifically in Leydig cells, blood vessels, spermatogonia and elongated spermatids), epididymis (specifically in the cytoplasm of the principal epithelial cells, luminal secretion and in the surrounding smooth muscle) and the lining epithelia of the accessory sexual glands (cytoplasmic location in the prostate and membranous in the seminal vesicle and bulbourethral glands). The Western blotting analysis confirmed the presence of PON1 in all boar genital organs, showing in all of them a band of 51 kDa and an extra band of 45 kDa only in seminal vesicles. PON1 showed higher activity levels in epididymal fluid than those in SP of the entire ejaculate or of specific ejaculate portions. A highly positive relationship between PON1 activity and HDL-C concentration was found in all genital fluids. In sum, all boar genital organs contributing to sperm-accompanying fluid/s were able to express PON1, whose activity in these genital fluids is highly dependent on the variable HDL-C concentration present.
Subject(s)
Aryldialkylphosphatase/metabolism , Semen/enzymology , Seminal Vesicles/enzymology , Spermatozoa/enzymology , Testis/enzymology , Animals , Male , SwineABSTRACT
A complete proteomic profile of seminal plasma (SP) remains challenging, particularly in porcine. The data reports on the analysis of boar SP-proteins by using a combination of SEC, 1-D SDS PAGE and NanoLC-ESI-MS/MS from 33 pooled SP-samples (11 boars, 3 ejaculates/boar). A complete dataset of the 536 SP-proteins identified and validated with confidence ≥95% (Unused Score >1.3) and a false discovery rate (FDR) ≤1%, is provided. In addition, the relative abundance of 432 of them is also shown. Gene ontology annotation of the complete SP-proteome complemented by an extensive description of the putative reproductive role of SP-proteins, providing a valuable source for a better understanding of SP role in the reproductive success. This data article refers to the article entitled "Characterization of the porcine seminal plasma proteome comparing ejaculate portions" (Perez-Patiño et al., 2016) [1].
ABSTRACT
Glutathione peroxidase-5 (GPX5) is an H2O2-scavenging enzyme identified in boar seminal plasma (SP). This study attempted to clarify its origin and role on sperm survival and fertility after artificial insemination (AI). GPX5 was expressed (Western blot and immunocytochemistry using a rabbit primary polyclonal antibody) in testes, epididymis and accessory sex glands (6 boars). SP-GPX5 concentration differed among boars (11 boars, P < 0.001), among ejaculates within boar (44 ejaculates, P < 0.001) and among portions within ejaculate (15 ejaculates). The first 10 mL of the sperm rich fraction (SRF, sperm-peak portion) had a significantly lower concentration (8.87 ± 0.78 ng/mL) than the rest of the SRF and the post-SRF (11.66 ± 0.79 and 12.37 ± 0.79 ng/mL, respectively, P < 0.005). Sperm motility of liquid-stored semen AI-doses (n = 44, at 15-17°C during 72h) declined faster in AI-doses with low concentrations of SP-GPX5 compared to those with high-levels. Boars (n = 11) with high SP-GPX5 showed higher farrowing rates and litter sizes than those with low SP-GPX5 (a total of 5,275 inseminated sows). In sum, GPX5 is widely expressed in the boar genital tract and its variable presence in SP shows a positive relationship with sperm quality and fertility outcomes of liquid-stored semen AI-doses.
Subject(s)
Genitalia, Male/enzymology , Glutathione Peroxidase/physiology , Semen/enzymology , Spermatozoa/enzymology , Swine/physiology , Animals , Fertility/physiology , Genitalia, Male/physiology , Glutathione Peroxidase/metabolism , Insemination, Artificial/veterinary , Male , Sperm Motility/physiology , Spermatozoa/physiologyABSTRACT
UNLABELLED: Full identification of boar seminal plasma (SP) proteins remains challenging. This study aims to provide an extensive proteomic analysis of boar SP and to generate an accessible database of boar SP-proteome. A SP-pool (33entire ejaculates/11 boars; 3ejaculates/boar) was analyzed to characterize the boar SP-proteome. Twenty ejaculates (5 boars, 4ejaculates/boar) collected in portions (P1: first 10mL of sperm rich ejaculate fraction (SRF), P2: rest of SRF and P3: post-SRF) were analyzed to evaluate differentially expressed SP-proteins among portions. SP-samples were analyzed using a combination of SEC, 1-D SDS PAGE and NanoLC-ESI-MS/MS followed by functional bioinformatics. The identified proteins were quantified from normalized LFQ intensity data. A total of 536 SP-proteins were identified, 409 of them in Sus scrofa taxonomy (374 validated with ≥99% confidence). Barely 20 of the identified SP-proteins were specifically implicated in reproductive processes, albeit other SP-proteins could be indirectly involved in functionality and fertility of boar spermatozoa. Thirty-four proteins (16 identified in S. scrofa taxonomy) were differentially expressed among ejaculate portions, 16 being over-expressed and 18 under-expressed in P1-P2 regarding to P3. This major proteome mapping of the boar SP provides a complex inventory of proteins with potential roles as sperm function- and fertility- biomarkers. BIOLOGICAL SIGNIFICANCE: This proteomic study provides the major characterization of the boar SP-proteome with >250 proteins first reported. The boar SP-proteome is described so that a spectral library can be built for relative 'label free' protein quantification with SWATH approach. This proteomic profiling allows the creation of a publicly accessible database of the boar SP-proteome, as a first step for further understanding the role of SP-proteins in reproductive outcomes as well as for the identification of biomarkers for sperm quality and fertility.
Subject(s)
Proteome/analysis , Proteomics/methods , Seminal Plasma Proteins/analysis , Animals , Chromatography, Liquid , Computational Biology , Databases, Protein , Ejaculation , Electrophoresis, Polyacrylamide Gel , Fertility , Gene Expression Regulation , Male , Reproduction , Semen Analysis , Swine , Tandem Mass SpectrometryABSTRACT
The study attempted to clarify the role of total antioxidant capacity of seminal plasma (SP-TAC) on boar sperm survival and fertility after artificial insemination (AI). SP-TAC differed (P < 0.001) among boars (n° = 15) and, to a lesser degree, among ejaculates within male (4 ejaculates/boar). SP-TAC also differed (P < 0.001) among ejaculate fractions (43 ejaculates and 3 fractions per ejaculate), of which the sperm-peak portion of the sperm rich ejaculate fraction (SRF) had the highest SP-TAC. SP-TAC was not correlated with sperm quality (motility and viability) or functionality (intracellular ROS generation and lipid peroxidation) of liquid AI-semen samples stored at 17 °C for 72 h (90 AI-samples), but the decline in sperm quality was larger (P < 0.05) in ejaculates with low, compared with high SP-TAC (hierarchically grouped). The SP-TAC differences among ejaculate portions agree with sperm cryosurvival rates (14 ejaculates from 7 boars), showing sperm from sperm-peak portion better (P < 0.01) post-thaw quality and functionality than those from the entire ejaculate (mainly post-SRF). Boars (n° = 18) with high SP-TAC (hierarchically grouped) had higher (P < 0.05) fertility outcomes (5,546 AI-sows) than those with low SP-TAC. Measurement of SP-TAC ought to be a discriminative tool to prognosis fertility in breeding boars.
Subject(s)
Antioxidants/metabolism , Fertility , Semen/metabolism , Spermatozoa/metabolism , Animals , Cell Survival , Insemination, Artificial , Male , Semen Analysis , Sperm Motility , SwineABSTRACT
PROBLEM: The boar, as human, sequentially ejaculates sperm-rich and sperm-poor fractions. Seminal plasma (SP) spermadhesins (PSP-I/PSP-II) induce a primary endometrial inflammatory response in female sows, similar to that elicited by semen deposition in other species, including human. However, the SP is also known to mitigate such response, making it transient to allow for embryo entry to a cleansed endometrium. Although cytokine involvement has been claimed, the exploration of cytokines in different SP fractions is scarce. This study determines Th1, Th2, Th17 and Th3 cytokine profiles in specific ejaculate SP fractions from boars of proven fertility. METHODS: SP samples from the sperm-rich fraction (SRF) and the sperm-poor post-SRF fraction (post-SRF) of manually collected ejaculates from eight boars (four ejaculates per boar) were analysed by commercial multiplex bead assay kits (Milliplex MAP, Millipore, USA) for interferon-γ, interferon gamma-induced protein 10, macrophage-derived chemokine, growth-regulated oncogene, granulocyte-macrophage colony-stimulating factor, monocyte chemo-attractant protein-1, interleukins (IL)-6, IL-8, IL-10, IL-15, IL-17 and transforming growth factor (TGF)-ß1-ß3. RESULTS: Cytokine concentrations differed between the ejaculate fractions among boars, being highest in the post-SRF. CONCLUSION: Boar SP is rich in Th1, Th2, Th17 and Th3 cytokines, with lowest concentrations in the sperm-peak-containing fraction, indicating its main immune influence might reside in the larger, protein-rich sperm-poor post-SRF.