Assesment of intracellular homocysteine concentration in sperms

a 2011

Assesment of intracellular homocysteine concentration in sperms

ŽÁKOVÁ, Jana; Michaela KRÁLÍKOVÁ; Igor CRHA; Pavel VENTRUBA; Jitka MELOUNOVÁ et al.

Základní údaje

Originální název

Assesment of intracellular homocysteine concentration in sperms

Autoři

ŽÁKOVÁ, Jana; Michaela KRÁLÍKOVÁ ; Igor CRHA; Pavel VENTRUBA; Jitka MELOUNOVÁ; Milena MATEJOVIČOVÁ; M. VODOVÁ a E. LOUSOVÁ

Vydání

Human Reproduction. 27th Annual Meeting of the European Society of Human Reproduction and Embryology ESHRE, 2011

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

30214 Obstetrics and gynaecology

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 4.475

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14110/11:00053108

Organizační jednotka

Lékařská fakulta

ISSN

UT WoS

000294450500331

Klíčová slova anglicky

intracellular homocysteine sperms

Příznaky

Mezinárodní význam

Změněno: 15. 9. 2011 12:47, Mgr. Michal Petr

Anotace

V originále

The aim of this study was to evolve an effective procedure for the sperm lysis, obtention of intracellular content and subsequent assesment of Hc concentration and, in consequence, determine the correlations between blood plasma and seminal fluid Hc concentration, spermiogram parameters and DNA injury. Firstly, we used the method of repeated freezing and melting according to Ebisch et al., 2006 for the sperm lysis. Because the microscopic control of the samples didn’t show any markable sperm injury, we projected and compared other methods of the cell lysis – ultrasound, melting in ultrasound and freezing 10 times, methanol, distilled water, Triton X-100 solution, and lysis solution + 1% Triton X-100 solution. We used the method of high-performance liquid chromatography with fluorescence detection (HPLCFD) for the determination of Hc concentration and comet analysis for the assesment of DNA injury. Correlation coefficient was used to determine the correlations between the intracellular Hc concentration and the other parameters. The most effective methods of sperm lysis were resuspending of sperms in distilled water or 80% methanol, one-hour incubation and subsequent fivefold freezing and melting in ultrasound bathe. Distilled water causes osmotic shock and membrane craze, methanol causes protein denaturation. Measured Hc concentrations ranged between 85.8–528.4 nmol/106 of sperms (median 233.4 nmol/106 sp., mean 265.4 nmol/106 sp., N = 8). We found positive correlation between intracellular concentration of Hc and rate of sperms with abnormal neck morphology (R = 0.818, p < 0.05) and weak positive correlation between seminal fluid and blood plasma Hc (R = 0.645 and R = 0.574, respectively). We measured the intracellular Hc concentrations which are three orders of magnitude higher than published. We attribute it to really effective breaking of sperms and getting intracellular content. The results show possible correlations with some spermiogram parameters.

Citovat

ŽÁKOVÁ, Jana; Michaela KRÁLÍKOVÁ; Igor CRHA; Pavel VENTRUBA; Jitka MELOUNOVÁ; Milena MATEJOVIČOVÁ; M. VODOVÁ a E. LOUSOVÁ. Assesment of intracellular homocysteine concentration in sperms. In Human Reproduction. 27th Annual Meeting of the European Society of Human Reproduction and Embryology ESHRE. 2011. ISSN 0268-1161.

@proceedings{950123,
   author = {Žáková, Jana and Králíková, Michaela and Crha, Igor and Ventruba, Pavel and Melounová, Jitka and Matejovičová, Milena and Vodová, M. and Lousová, E.},
   booktitle = {Human Reproduction. 27th Annual Meeting of the European Society of Human Reproduction and Embryology ESHRE},
   keywords = {intracellular homocysteine sperms},
   language = {eng},
   title = {Assesment of intracellular homocysteine concentration in sperms},
   year = {2011}
}

TY  - CONF
ID  - 950123
AU  - Žáková, Jana - Králíková, Michaela - Crha, Igor - Ventruba, Pavel - Melounová, Jitka - Matejovičová, Milena - Vodová, M. - Lousová, E.
PY  - 2011
TI  - Assesment of intracellular homocysteine concentration in sperms
KW  - intracellular homocysteine sperms
N2  - The aim of this study was to evolve an effective procedure for the sperm lysis, obtention of intracellular content and subsequent assesment of Hc concentration and, in consequence, determine the correlations between blood plasma and seminal fluid Hc concentration, spermiogram parameters and DNA injury. Firstly, we used the method of repeated freezing and melting according to Ebisch et al., 2006 for the sperm lysis. Because the microscopic control of the samples didn’t show any markable sperm injury, we projected and compared other methods of the cell lysis – ultrasound, melting in ultrasound and freezing 10 times, methanol, distilled water, Triton X-100 solution, and lysis solution + 1% Triton X-100 solution. We used the method of high-performance liquid chromatography with fluorescence detection (HPLCFD) for the determination of Hc concentration and comet analysis for the assesment of DNA injury. Correlation coefficient was used to determine the correlations between the intracellular Hc concentration and the other parameters. The most effective methods of sperm lysis were resuspending of sperms in distilled water or 80% methanol, one-hour incubation and subsequent fivefold freezing and melting in ultrasound bathe. Distilled water causes osmotic shock and membrane craze, methanol causes protein denaturation. Measured Hc concentrations ranged between 85.8–528.4 nmol/106 of sperms (median 233.4 nmol/106 sp., mean 265.4 nmol/106 sp., N = 8). We found positive correlation between intracellular concentration of Hc and rate of sperms with abnormal neck morphology (R = 0.818, p < 0.05) and weak positive correlation between seminal fluid and blood plasma Hc (R = 0.645 and R = 0.574, respectively). We measured the intracellular Hc concentrations which are three orders of magnitude higher than published. We attribute it to really effective breaking of sperms and getting intracellular content. The results show possible correlations with some spermiogram parameters.
ER  -

ŽÁKOVÁ, Jana; Michaela KRÁLÍKOVÁ; Igor CRHA; Pavel VENTRUBA; Jitka MELOUNOVÁ; Milena MATEJOVIČOVÁ; M. VODOVÁ a E. LOUSOVÁ. Assesment of intracellular homocysteine concentration in sperms. In \textit{Human Reproduction. 27th Annual Meeting of the European Society of Human Reproduction and Embryology ESHRE}. 2011. ISSN~0268-1161.

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