Seminal lipid profiling and antioxidant capacity : a species comparison

On their way to the oocyte, sperm cells are subjected to oxidative stress, which may trigger the oxidation of phospholipids (PL). Applying MALDI-TOF MS, HPTLC and ESI-IT MS, we comparatively analyzed the PL compositions of semen and blood of species differing in their reproductive systems and types of nutrition (bull, boar, stallion, lion and man) with regard to the sensitivity to oxidation as well as the accumulation of harmful lyso-PL (LPL), transient products of lipid oxidation. In addition, the protective capacity of seminal fluid (SF) was also examined. The PL composition of erythrocytes and blood plasma is similar across the species, while pronounced differences exist for sperm and SF. Since the blood function is largely conserved across mammalian species, but the reproductive systems may vary in many aspects, the obtained results suggest that the PL composition is not determined by the type of nutrition, but by the relatedness of species and by functional requirements of cell membranes such as fluidity. Sperm motion and fertilization of oocytes require a rather flexible membrane, which is accomplished by significant moieties of unsaturated fatty acyl residues in sperm lipids of most species, but implies a higher risk of oxidation. Due to a high content of plasmalogens (alkenyl ether lipids), bull sperm are most susceptible to oxidation. Our data indicate that bull sperm possess the most effective protective power in SF. Obviously, a co-evolution of PL composition and protective mechanisms has occurred in semen and is related to the reproductive characteristics. Although the protective capacity in human SF seems well developed, we recorded the most pronounced individual contaminations with LPL in human semen. Probably, massive oxidative challenges related to lifestyle factors interfere with natural conditions.SUPPLEMENTARY MATERIAL: S1 Fig. ESI spectra of lysophosphatidylcholine (LPC) fractions from boar, bull, stallion, lion and human samples.S2 Fig. ESI spectra of sphingomyelin (SM) fractions from boar, bull, stallion, lion and human samples. Lipid extracts were separated on a normal phase high performance thin-layer chromatography (HPTLC) plate with chloroform/ethanol/water/triethylamine (30:35:7:35, by vol.) as the mobile phase. Plates were air-dried and stained with primuline (Direct Yellow 59, Sigma-Aldrich, Taufkirchen, Germany) (50 mg/l dissolved in acetone/water 80:20, by vol.). Lipids were made visible under UV light and marked with a pencil. SM fractions were directly analyzed by coupling a TLC plate reader to an ESI mass spectrometer. Mass spectra were recorded in the positive ion mode. For further details on ESI-IT MS see main text. For peak assignment, please see S2 Table. https://doi.org/10.1371/journal.pone.0264675.s002S3 Fig. ESI spectra of phosphatidylcholine (PC) fractions from boar, bull, stallion, lion and human samples. Lipid extracts were separated on a normal phase high performance thin-layer chromatography (HPTLC) plate with chloroform/ethanol/water/triethylamine (30:35:7:35, by vol.) as the mobile phase. Plates were air-dried and stained with primuline (Direct Yellow 59, Sigma-Aldrich, Taufkirchen, Germany) (50 mg/l dissolved in acetone/water 80:20, by vol.). Lipids were made visible under UV light and marked with a pencil. PC fractions were directly analyzed by coupling a TLC plate reader to an ESI mass spectrometer. Mass spectra were recorded in the positive ion mode. For further details on ESI-IT MS see main text. For peak assignment, please see S3 Table. https://doi.org/10.1371/journal.pone.0264675.s003S4 Fig. ESI spectra of phosphatidylinositol (PI) fractions from boar, bull, stallion and human lipid samples. Lipid extracts were separated on a normal phase high performance thin-layer chromatography (HPTLC) plate with chloroform/ethanol/water/triethylamine (30:35:7:35, by vol.) as the mobile phase. Plates were air-dried and stained with primuline (Direct Yellow 59, Sigma-Aldrich, Taufkirchen, Germany) (50 mg/l dissolved in acetone/water 80:20, by vol.). Lipids were made visible under UV light and marked with a pencil. PI fractions were directly analyzed by coupling a TLC plate reader to an ESI mass spectrometer. Mass spectra were recorded in the negative ion mode. For further details on ESI-IT MS see main text. For peak assignment, please see S4 Table. https://doi.org/10.1371/journal.pone.0264675.s004S5 Fig. ESI spectra of phosphatidylethanolamine (PE) fractions from boar, bull and stallion samples. Lipid extracts were separated on a normal phase high performance thin-layer chromatography (HPTLC) plate with chloroform/ethanol/water/triethylamine (30:35:7:35, by vol.) as the mobile phase. Plates were air-dried and stained with primuline (Direct Yellow 59, Sigma-Aldrich, Taufkirchen, Germany) (50 mg/l dissolved in acetone/water 80:20, by vol.). Lipids were made visible under UV light and marked with a pencil. PE fractions were directly analyzed by coupling a TLC plate reader to an ESI mass spectrometer. Mass spectra were recorded in the negative ion mode. For further details on ESI-IT MS see main text. For peak assignment, please see S5 Table. https://doi.org/10.1371/journal.pone.0264675.s005S6 Fig. ESI spectra of phosphatidylethanolamine (PE) fractions from lion and human samples. Lipid extracts were separated on a normal phase high performance thin-layer chromatography (HPTLC) plate with chloroform/ethanol/water/triethylamine (30:35:7:35, by vol.) as the mobile phase. Plates were air-dried and stained with primuline (Direct Yellow 59, Sigma-Aldrich, Taufkirchen, Germany) (50 mg/l dissolved in acetone/water 80:20, by vol.). Lipids were made visible under UV light and marked with a pencil. PE fractions were directly analyzed by coupling a TLC plate reader to an ESI mass spectrometer. Mass spectra were recorded in the negative ion mode. For further details on ESI-IT MS see main text. For peak assignment, please see S5 Table. https://doi.org/10.1371/journal.pone.0264675.s006S7 Fig. Hydrolysis of selected seminal fluid samples over time. The plots of hydrolysis measurements from boar and stallion seminal fluid were fitted by a linear curve (f(x) = a + b×x) and the plots from bull, lion and human were fitted by an exponential growth to a maximum (f(x) = a×e-b×x). Due to these different courses of the hydrolysis reaction between the species, the absolute hydrolysis at a given time point (10 min) was used to compare the mean values of the investigated individuals between the species (see Table 2 of the main text). https://doi.org/10.1371/journal.pone.0264675.s007S8 Fig. Effect of artificial LPC on boar sperm. Beltsville Thawing Solution (BTS, Minitüb GmbH)-diluted boar semen (20 × 106 sperm/ml) was mixed with 20 μM lysophosphatidylcholine (LPC 16:0, Avanti Polar Lipids®, No 855675C). After incubation at 38°C for 30 min, the ratios of total motility (blank boxes) and sperm with an intact acrosome (striped boxes) were analyzed. The lipid extract of washed sperm of this experiment was analyzed by MALDI-TOF MS and the ratio of LPC to total GPC was calculated (for details see Material and Methods of the main text). Incubation with 20 μM LPC led to 2.4 ± 3.6% inserted LPC in sperm cell membranes. Significant differences in total motility and the percentage of sperm with an intact acrosome between the incubation with 20 μM LPC and controls are marked by asterisks (P = 0.006 and 0.003, respectively, Wilcoxon signed-rank test, n = 11). https://doi.org/10.1371/journal.pone.0264675.s008S9 Fig. Original TLC pictures. Lipid extracts were separated on normal phase high performance thin-layer chromatography (HPTLC) plates with chloroform/ethanol/water/triethylamine (30:35:7:35, by vol.) as the mobile phase. Plates were air-dried and stained with primuline (Direct Yellow 59, Sigma-Aldrich, Taufkirchen, Germany) (50 mg/l dissolved in acetone/water 80:20, by vol.). BP–blood plasma, SF–seminal fluid, st.–lipid standard mixture made of LPC16:0, SM16:0, PC16:0/18:1, PA 16:0/18:1, PI 16:1/18:1, PE 16:0/18:1, PG 16:0/18:1 (bottom up). https://doi.org/10.1371/journal.pone.0264675.s009S1 Table. Assignment of signals detected in ESI spectra from lysophosphatidylcholine (LPC) spots. https://doi.org/10.1371/journal.pone.0264675.s010S2 Table. Assignment of signals detected in ESI spectra from sphingomyelin (SM) spots. n.a.—not assigned. https://doi.org/10.1371/journal.pone.0264675.s011S3 Table. Assignment of signals detected in ESI spectra from phosphatidylcholine (PC) spots. https://doi.org/10.1371/journal.pone.0264675.s012S4 Table. Assignment of signals detected in ESI spectra from phosphatidylinositol (PI) spots. https://doi.org/10.1371/journal.pone.0264675.s013S5 Table. Assignment of signals detected in ESI spectra from phosphatidylethanolamine (PE) spots. https://doi.org/10.1371/journal.pone.0264675.s014The German Research Council.http://www.plosone.orgdm2022Veterinary Tropical Disease

An adjoint method for the assimilation of statistical characteristics into eddy-resolving ocean models

The study investigates perspectives of the parameter estimation problem with the adjoint method in eddy-resolving models. Sensitivity to initial conditions resulting from the chaotic nature of this type of model limits the direct application of the adjoint method by predictability. Prolonging the period of assimilation is accompanied by the appearance of an increasing number of secondary minima of the cost function that prevents the convergence of this method. In the framework of the Lorenz model it is shown that averaged quantities are suitable for describing invariant properties, and that secondary minima are for this type of data transformed into stochastic deviations. An adjoint method suitable for the assimilation of statistical characteristics of data and applicable on time scales beyond the predictability limit is presented. The approach assumes a greater predictability for averaged quantities. The adjoint to a prognostic model for statistical moments is employed for calculating cost function gradients that ignore the fine structure resulting from secondary minima. Coarse resolution versions of eddy-resolving models are used for this purpose. Identical twin experiments are performed with a quasigeostrophic model to evaluate the performance and limitations of this approach in improving models by estimating parameters. The wind stress curl is estimated from a simulated mean stream function. A very simple parameterization scheme for the assimilation of second-order moments is shown to permit the estimation of gradients that perform efficiently in minimizing cost functions

Investigations of the lysophospholipid composition of human neutrophils under different stimulation conditions by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS) is usually used for the analyses of proteins, carbohydrates and oligonucleotides. In spite of the number of advantages that MALDI-TOF MS exhibits for lipid analysis, this method has not often been applied in this field. In this paper we have extended our previous studies on the suitability of MALDI-TOF MS for the investigation of changes in the content of lipid-derived second messengers in organic extracts of human neutrophils. Qualitative differences in the lysophospholipid composition in organic extracts of the human neutrophils under different stimulation conditions could be easily observed by MALDI-TOF MS. Although there are still some methodological problems to be solved before this method can be routinely applied for the quantification of different lipid classes in complex biological mixtures (such as organic extracts of human neutrophils) it is shown here that MALDI-TOF MS possesses the capability to be used as a simple screening method for the investigation of the content of lipid-derived second messengers and of signalling pathways in cells