Parental sex effect of parthenogenesis on hatchability and sperm-egg penetration in mated Chinese Painted quail (Coturnix chinensis).

Selecting quail for an increased incidence of parthenogenesis also impacts egg weight and albumen pH as well as reduces hatchability and fertility due to decreased sperm-egg penetration (SEP). However, it is unknown which parental sex is responsible for these changes in quail selected for parthenogenesis. Therefore, the objective of this study was to determine which sex influences egg weight, albumen pH, hatchability, and SEP in birds selected for parthenogenesis. In this study, 2 lines of birds were used: 1 line that was selected for parthenogenesis and 1 line not selected for parthenogenesis (control). Treatments were as follows: control females w/control males, control females w/parthenogenetic line males, parthenogenetic line females w/control males, and parthenogenetic line females w/parthenogenetic line males. Fresh eggs were collected daily, labeled and analyzed for albumen pH and SEP or incubated at 37.5 °C for 20 d of incubation. Eggs were candled at 10 days of incubation (DOI) and eggs exhibiting little or no embryonic development were removed and broken open to determine hatching failure. This was repeated at 20 DOI for eggs that did not hatch. A dam main effect for egg set weight existed with parthenogenetic line dams exhibiting heavier eggs than control dams. The parthenogenetic line dams and sires exhibited lower albumen pH and hatch but a higher incidence of parthenogenesis than control line dams or sires. However, only a sire main effect existed for fertility and SEP. Sires from the parthenogenetic line yielded the highest infertility due to lower SEP. In conclusion, both the parthenogenetic line dams and sires contribute to reduced reproductive performance. However, it appears that the sire from the parthenogenetic line is responsible for lower fertility due to a reduction in SEP. Because the sire has a negative impact on overall fertility, it is possible that males selected for parthenogenesis have poorer semen quality resulting in fewer sperm traversing the oviduct or penetrating the perivitelline layer.

Lipid peroxidation of circulating low density lipoproteins with age, smoking and in peripheral vascular disease.

In this study, lipid peroxides in plasma and the low density lipoprotein (LDL) fraction and plasma concentrations of vitamin E, lipids and lipoproteins were measured in 22 smokers (mean age 35 years), 26 non-smoking patients with peripheral vascular disease (PVD), mean age 66 years), and 23 younger (ages < or = 55 years) and 26 older (ages > 55 years) healthy subjects. Plasma lipid peroxide concentrations in the PVD patients (105.9 +/- 20.6 vs. 91.8 +/- 15.8 ng malondialdehyde (MDA)/ml plasma, mean +/- S.D.) and the smokers (94.1 +/- vs. 74.0 +/- 13.9 ng MDA/ml plasma) were significantly elevated compared with levels in the appropriate control subjects and levels were significantly higher in older compared with younger control subjects. Plasma LDL lipid peroxides were also significantly raised in patients with PVD and smokers compared with control values (PVD): 37.1 +/- 7.7 vs. 26.3 +/- 4.1 ng MDA/ml plasma; smokers: 30.4 +/- 6.9 vs 24.9 +/- 7.5 ng MDA/ml plasma). The ratio of LDL lipid peroxides: LDL-cholesterol was significantly higher in the smokers, and plasma cholesterol and LDL-cholesterol were significantly higher in patients with PVD compared with other groups of subjects. The ratio of vitamin E: total lipid was not significantly different between the study groups. These data show that lipid peroxide levels in the plasma LDL fraction are elevated along with raised circulating levels in patients with PVD and smokers but that LDL lipid peroxide concentrations were independent of age in the healthy subjects. Elevated LDL lipid peroxide concentrations may may be mainly due to abnormally high LDL levels in PVD patients, whereas in smokers, the concentration of lipid peroxides in the LDL particles is raised and might render the lipoprotein more atherogenic.

Selenium and vitamin E supplementation: activities of glutathione peroxidase in human tissues.

Twenty-seven New Zealand women received daily for 4 wk, 200 micrograms selenium as sodium selenite, 170 mg alpha-tocopherol acetate, or a placebo. Se supplementation raised platelet selenoglutathione peroxidase (Se-GSHPx, p less than 0.001) and also Se and Se-GSHPx in whole blood and plasma. Se concentrations and Se-GSHPx activities in liver biopsies taken after supplementation were greater (p less than 0.05) for the Se group and a good correlation was found between Se and Se-GSHPx in liver and muscle for all subjects. Platelet Se-GSHPx correlated well with Se and Se-GSHPx in liver, indicating its suitability for assessing Se bioavailability. This is the first reported study of relationships between Se and Se-GSHPx in human liver and muscle tissue and platelet Se-GSHPx after Se supplementation. These observations verify in man relationships observe in animal studies, giving support to assumptions made in methods for assessing Se status and bioavailability in man, in particular the use of platelet GSHPx.

The absence of adaptive changes in tissue activities of glutathione-S-transferase, superoxide dismutase and catalase following selenium and vitamin E supplementation in subjects with low selenium status.

Three groups of New Zealand women were given daily in a double blind randomised study, 200 micrograms Se as sodium selenite, 170 mg alpha-tocopherol or a placebo for 4 wk. Activities of glutathione-S-transferase, superoxide dismutase and catalase were assayed in erythrocytes, plasma and platelets and in liver and muscle biopsy tissues. No changes in activities of any of these tissue enzymes were observed in any of the three groups. There were also no changes in non-selenium dependent glutathione peroxidase activities in liver or plasma. The lack of changes in any of these enzymes following selenium supplementation suggests that adaptive changes to the low selenium status of these subjects had not occurred through these lipid peroxidation defense mechanisms.

Plasma thiobarbituric acid reactivity: reaction conditions and the role of iron, antioxidants and lipid peroxy radicals on the quantitation of plasma lipid peroxides.

The effects of Fe3+, lipid peroxy radicals and the antioxidant butylated hydroxytoluene on the 2-thiobarbituric (TBA) acid quantitation of plasma lipid peroxides were investigated. Whole plasma and plasma fractions prepared by trichloroacetic acid (TCA) protein precipitation and lipid extraction, demonstrated markedly differing TBA reactivities in the presence or absence of added Fe3+. Examination of the spectral profiles of the TBA reacted whole plasma and TCA precipitated fractions demonstrated the presence of interfering compounds which gave rise to an artifactual increase in lipid peroxide concentrations. In contrast the TBA reacted lipid extracts had low levels of interfering compounds that could be removed by our previously described high pressure liquid chromatographic method (Wade, Jackson and van Rij (1985) Biochem. Med. 33, 291-296). Further characterization of the TBA reactivity of the lipid extract showed that Fe3+ at an optimal concentration of 0.5 mM was necessary for the quantitative decomposition of the lipid peroxides to the TBA reactive product malondialdehyde (MDA). However the presence of Fe3+ resulted in further peroxidation of any unsaturated lipids present. Butylated hydroxytoluene (BHT) at an optimal concentration of 1.4 mM inhibited Fe3+ stimulated peroxidation without affecting the formation of the MDA-TBA chromogen. Using a standardized TBA test with plasma lipid extracts and the addition of optimal concentrations of Fe3+ and BHT, we have determined the mean concentration of lipid peroxides in 30 healthy human subjects to be 102.7 +/- 20.0 ngm/ml.

Lipid peroxidation and malondialdehyde in the synovial fluid and plasma of patients with rheumatoid arthritis.

The concentration of lipid peroxides in the plasma and synovial fluid of 65 arthritic patients was determined using a new ion-pairing reverse phase HPLC technique. Patients with rheumatoid arthritis receiving only non-steroidal anti-inflammatory drugs, had a significantly higher mean concentration of lipid peroxides in synovial fluid samples (162 +/- 22.0 micrograms/l) than osteoarthritic patients (40.0 +/- 8.0 micrograms/l, p less than 0.0001). Mean concentrations in both groups correlated strongly with the level of beta-glucuronidase activity as a measure of lysosomal enzyme release (r = 0.71, p less than 0.0001). Contrary to previous reports by investigators using less specific methods, we were unable to demonstrate any increase in plasma levels of lipid peroxides in the rheumatoid patient. Treatment of rheumatoid arthritis with D-penicillamine was associated with a significant reduction of lipid peroxide levels (83.2 +/- 11.5 micrograms/ml, p less than 0.002), suggesting that this drug may function as an oxygen radical scavenger in the joint cavity. These results give further support to the concept of oxygen-free radicals playing an important role in the pathogenesis of chronic inflammatory disorders.

The metabolism of low molecular weight hydrocarbon gases in man.

The metabolism of ethane and pentane in man is demonstrated to occur from the uptake of an enriched atmosphere of these gases in a rebreathe spirometer circuit. Dithiocarb, an inhibitor of alkane metabolism, reduced uptake and increased the respiratory excretion of these gases. This effect was least marked for the slowly metabolised ethane. Therefore the endogenous production of ethane as measured by respiratory excretion is less affected. However pentane is rapidly metabolised and this limits the use of simple respiratory excretion of pentane as a measure of in vivo lipid peroxidation.

In vivo lipid peroxidation in man as measured by the respiratory excretion of ethane, pentane, and other low-molecular-weight hydrocarbons.

A method for the collection and measurement of low-molecular-weight volatile hydrocarbons exhaled in human breath as a result of lipid peroxidation in vivo is described. Subjects breathed in a closed-circuit rebreathe system and samples were taken over a 2-h period, extracted, and analyzed by gas-liquid chromatography isothermally on a 3-m column of n-octane/Porasil-C. Immediately, prior to rebreathing subjects were equilibrated with scrubbed air containing very low ambient levels of hydrocarbons to eliminate the effects of previous exposure to hydrocarbon-contaminated environments. Only under these conditions could hydrocarbon exhalation be measured. In the rebreathe system C3-C5 hydrocarbon concentrations increased linearly initially but reached a steady state after 1.5 h while ethane did not approach equilibrium even after 2 h. The steady-state equilibrium was demonstrated to be due to tissue uptake and metabolism of the hydrocarbon gases. Ethane metabolism was slow, allowing calculation of the endogenous production from the initial rate of change in concentration and an experimentally determined total body solubility coefficient. Similar calculations for pentane were not valid as metabolism was rapid; therefore production was estimated from the equilibrium value reached after 2 h. Ethane production in six healthy subjects was calculated to be 95.1 +/- 19.0 pmol/kg/h while equilibrium values for pentane were 120 +/- 50 pmol/liter. This method now allows the quantitation in man of lipid peroxidation in vivo.

Quantitation of malondialdehyde (MDA) in plasma, by ion-pairing reverse phase high performance liquid chromatography.

An ion-pairing high performance liquid chromatography method is described for the separation and quantitation of malondialdehyde in plasma. The MDA is determined as the thiobarbiturate chromogen formed by reaction of the plasma with 2-thiobarbituric acid under acid and heating conditions. However, under these conditions other interfering chromogens can also be formed. Using DEAE-cellulose chromatography followed by ion-pairing HPLC, we have been able to separate and quantitate the levels of MDA-TBA chromogen formed in plasma from other interfering chromogens. Measurements of MDA levels in the plasma of six normal individuals by HPLC gives a mean value of 4.57 +/- 0.33 nmole/ml, whereas the spectrophotometric determined value is 8.83 +/- 1.15 nmole/ml. These data suggest that some reevaluation of the numerous papers published on MDA levels in plasma using spectrophotometric methods may be necessary.