RESUMO
Long-term exposure of liquid preserved boar spermatozoa to seminal plasma (SP) can cause dramatic sperm injury. This study examined whether boar specificity exists in the sensitivity of spermatozoa to SP and whether correspondent biomarkers can be identified. Consecutive ejaculates (n = 4-5) collected from 19 boars were centrifuged, diluted with a pH-stablising extender with 10% (v/v) autologous SP and evaluated by computer-assisted semen analysis and flow cytometry. Up until 144 h storage, four boars showed consistently high sperm motility, viability and mitochondria activity, and one boar showed consistently low values. Intra-boar variability was high in the other boars. Screening of SP (n = 12 samples) for protein markers using mass spectrometry identified three protein candidates of which the granulin precursor, legumain and AWN were 0.5 to 0.9 log2-fold less abundant (p < 0.05) in SP-resistant compared to SP-sensitive samples. Lipidome analysis by mass spectrometry revealed 568 lipids showing no difference between the SP-groups. The most abundant lipids were cholesterol (42,442 pmol), followed by phosphatidylserine (20,956 pmol) and ether-linked phosphatidylethanolamine (13,039 pmol). In conclusion, three candidate proteins were identified which might be indicative of SP-tolerance of sperm during long-term storage. Noteworthy, a first lipidomic profile of boar SP is presented.
Assuntos
Preservação do Sêmen/métodos , Sêmen/química , Espermatozoides/citologia , Animais , Criopreservação/métodos , Citometria de Fluxo , Lipidômica , Masculino , Proteínas/análise , Proteômica , Sêmen/citologia , Análise do Sêmen , Manejo de Espécimes/métodos , Motilidade dos Espermatozoides , Sus scrofa/metabolismo , SuínosRESUMO
Ongoing progress in proteomic characterization of seminal plasma has stimulated research on the identification of biomarkers for male fertility and sperm preservability. So far, many studies have evaluated the benefits of reconstituting cryopreserved or sex-sorted semen with seminal plasma. Less information is available about the effect of remaining or added seminal plasma in liquid preserved semen. The interaction between seminal plasma and spermatozoa is species -specific, and within species often complex and ambiguous. This article aims to review the action of seminal plasma on sperm function in preserved semen with a focus on liquid storage. Effects of seminal plasma on sperm traits during in vitro storage are summarized for males from four domestic farm animals, namely the bull, ram, boar and stallion. Special emphasis is placed on the effect of seminal plasma on long-term stored boar semen, including novel data demonstrating the attenuating effect of protective extender on the adverse effect of seminal plasma in some boars.
Assuntos
Preservação do Sêmen/veterinária , Sêmen/fisiologia , Espermatozoides/fisiologia , Animais , Bovinos , Cavalos , Masculino , Preservação do Sêmen/métodos , Preservação do Sêmen/normas , Ovinos , Especificidade da Espécie , Sus scrofaRESUMO
AIM: To collect data on the use of placebo interventions by GPs in Germany. METHODS: A questionnaire was mailed to 400 randomly selected GPs in Bavaria. Non-responders were reminded by telephone after 4 weeks and were given a second copy of the questionnaire after a further 3 weeks. RESULTS: In all, 208 completed questionnaires were returned. The majority of GPs (88%) have used a placebo at least once in their practice; 45% have used pure placebos, such as saline injections and sugar pills, at least once last year; the median frequency of use was 5 [interquartile range (IQR), 2-10]. The use of impure placebos during the past year was more common: 76% of GPs have used impure placebos, i.e. medical interventions that have pharmacological or physical activity but have no intrinsic effect (e.g. pharmacological or physical action) on the patient's disease or its symptoms, with a median frequency of 20 times per year (IQR, 10-50). The main reason for the use of placebo was a possible psychological effect, followed by the expectation of patients to receive a treatment. For the majority of GPs placebo interventions were ethically justified if they were used for a possible psychological effect. CONCLUSIONS: Placebo interventions are a widely accepted part of medical treatment in German general practices and are used primarily for their psychological effects. Impure placebos are used much more frequently than pure placebos.