Influence of two different feeding strategies in the dry period on dry matter intake and plasma protein peroxidative and antioxidative profile during dry period and early lactation.

BACKGROUND: Dairy cows undergo dramatic changes in endocrine and metabolic status around parturition and in early lactation. Meeting the nutritional requirements of transition dairy cows is important for animal health, production and animal wellbeing. Dry cow feeding and managing play an essential role in this. The changes in metabolism of periparturient cows also lead to a rise in the production of oxidising agents, leading to oxidative stress. The relationship between dry cow diet composition and oxidative stress has received little research attention so far. In the present study, the influence of two different dry cow feedings (single diet with medium energy content over the whole dry period versus traditional two-phase diet with a low-energy "far-off" ration and a high energy "close-up" ration) on dry matter intake, energy intake and plasma protein peroxidative and antioxidative profile was investigated. RESULTS: The examined parameters revealed a dynamic profile within the experimental period. Dry matter intake (DMI) did not differ between groups. However, there was a time and a group x time interaction effect: Group 1 ("one-phase") had a very constant DMI with a slow and even decrease until calving. In Group 2 ("two-phase"), an initial increase in DMI two weeks antepartum (a.p.) was followed by a sharp drop at week 1 a.p.. The highest total antioxidant capacity and sulfhydryl residue concentration was noted at partus. In contrast, concentration of formylokinurenine and bityrosine bridges as representatives of protein peroxidation were lowest at parturition. The time course of formylokinurenine and bityrosine bridges showed parallels to the DMI. The contents of sulfhydryl groups, formylokinurenine and total antixoxidant capacity did not differ between groups. In contrast, concentration of bityrosine bridges was always higher in Group 2 compared with Group 1 and these differences were statistically significant at week 3 a.p., week 2 a.p., week 1 a.p. and at parturition. CONCLUSION: The results of our study suggest time-related changes of pro- and antioxidative plasma parameters. Different dry cow feeding affected antepartal DMI. Furthermore, DMI and diet compositions seemed to have an influence on plasma protein peroxidative profile and activity of antioxidative defence.

2D Electrophoretic pattern of bovine placental proteins during early-mid pregnancy.

The Placenta, like every tissue, possesses its own characteristic protein profile, which may change within the course of pregnancy. These changes can be used for the elucidation of the mechanisms related to both physiology of pregnancy and pathological events. The aim of the study was to describe proteinergic profiles of maternal and fetal parts of bovine placenta during early-mid pregnancy by the use of 2D electrophoresis and MALDI TOF/TOF MS identification to evaluate dynamics of the possible changes necessary for placentation. Placental samples were collected from six pregnant cows (3-5 months) in the local abattoir. Placentomes were separated, and proteins were extracted and subjected to 2D electrophoresis and MALDI TOF/TOF identification. Out of 907 spots identified by the statistical analysis of gels, 54 were identified. Out of this number, 36 spots were significantly different between examined samples. Moreover, the obtained patterns differed between maternal and fetal parts of the placenta with regard to the intensity of staining, suggesting quantitative differences in protein content. These preliminary results are unique for this period of pregnancy. Such data are important for further experiments to obtain full protein profiles necessary to understand biochemical mechanisms underlying the attachment between fetal and maternal parts of the placenta during placentation. Moreover, the outcomes may help in elucidating pregnancy biomarkers in the future.

Patterns of protein glycosylation in bovine placentomes as a function of gestational age and in retained versus non-retained placenta.

The formation of placenta at the beginning of pregnancy and its separation at parturition require not only deep remodelling of extracellular matrix, which mainly consists of proteins conjugated with sugar moieties, but also the cooperation with cells from both maternal and foetal parts of placenta. The aim of the study was to compare the patterns of selected conjugated proteins with sugar moieties between pregnant and term placenta as well as between released and retained placenta in cows. Placental samples from healthy pregnant cows (3-5 months of pregnancy) were collected at a slaughterhouse (n = 6), and parturient samples were collected during caesarean section at term and retrospectively divided into retained (n = 6) and released (n = 6). The pattern of selected sugar moieties conjugated with proteins was detected by use of lectin blotting with Phaseolus Vulgaris leucoagglutinin, Maackia Amurensis and Sambucus Nigra (Elderberry). The comparison and analysis of obtained band patterns showed differences between their number, molecular weight and abundance related to the intensity of staining. Samples from 3 to 4 months showed similarities, while at the 5th month, clear differences were visible in all 3 lectins, which were used in this study. Samples from retained/released placenta expressed significant differences in PHA-L and SNA pattern in the foetal part. Obtained results indicate that the development of placenta related to extracellular matrix and accompanying cells from both sides of placenta shows dynamic changes during pregnancy. Moreover, in the case of animals with the retention of foetal membranes the patterns of proteins conjugated with sugar moieties are altered, suggesting that the changes in extracellular matrix metabolism can be involved in the attachment and detachment of the placenta in cows.

Biochemical Properties of Polyphenol Oxidases from Ready-to-Eat Lentil (Lens culinaris Medik.) Sprouts and Factors Affecting Their Activities: A Search for Potent Tools Limiting Enzymatic Browning.

Enzymatic browning of sprouts during storage is a serious problem negatively influencing their consumer quality. Identifying and understanding the mechanism of inhibition of polyphenol oxidases (PPOs) in lentil sprouts may offer inexpensive alternatives to prevent browning. This study focused on the biochemical characteristics of PPOs from stored lentil sprouts, providing data that may be directly implemented in improving the consumer quality of sprouts. The purification resulted in approximately 25-fold enrichment of two PPO isoenzymes (PPO I and PPO II). The optimum pH for total PPOs, as well as for PPO I and PPO II isoenzymes, was 4.5-5.5, 4.5-5.0, and 5.5, respectively. The optimal temperature for PPOs was 35 °C. Total PPOs and the PPO I and PPO II isoenzymes had the greatest affinity for catechol (Km = 1.32, 1.76, and 0.94 mM, respectively). Ascorbic acid was the most effective in the inhibition of dark color formation by total PPOs, and showed ca. 62%, 43%, and 24% inhibition at 20-, 2-, and 0.2-mM concentrations. Ascorbic acid, l-cysteine, and sodium metabisulfite (20 mM) significantly inhibited color development in the reactions catalyzed by both isoenzymes of PPO. Ba2+, Fe3+, and Mn2+ (10 mM) completely inhibited PPO activity. This study of the effect of antibrowning compounds and cations on PPO activity provides data that can be used to protect lentil sprouts against enzymatic browning during storage and processing.

The comparison of protein map between retained and released bovine placenta.

Placental retention in cows may be the result of altered protein pattern in comparison to physiologically released fetal membranes. Aim of study was to separate and identify proteins from maternal and fetal part of placenta and to compare them between released and retained fetal membranes. Six not retained and 6 retained tissues were obtained from healthy cows during routinely performed caesarian section. Cows were allocated to appropriate groups retrospectively. Samples were homogenized in phosphate buffer and subjected to 2D electrophoresis. After analysis of gels selected spots were excised and proteins were identified by MS. Two-dimensional electrophoresis detected and identified 886 spots in examined tissues. Significant differences (p < .05) were noticed between appropriate parts of retained and released placenta. In maternal part of retained placenta 40 spots showed lower abundance and 47 higher abundance in comparison to healthy samples. While in fetal part of retained placenta respective values were 60 and 125 proteins. Out of 73 identified proteins, 26 were significantly different between respective maternal (19) and fetal (7) part of retained and released placenta. In summary, protein profile of released and retained placenta express the presence and abundance of different proteins. It may suggest that selected proteins could be target molecules in searching for reasons for placental retention. Further identification of spots obtained here may provide with more detailed explanation of mechanisms of placental retention.

Identification of protein patterns in bovine placenta at early-mid pregnancy - Pilot studies.

RATIONALE: Placenta is a crucial tissue for an appropriate development of the fetus and the course of pregnancy. Its composition and structure change dynamically along pregnancy but the full pattern of these changes is not fully described in cows yet. The aim of the present study was to detect qualitative and quantitative protein profiles of bovine placenta during early-mid pregnancy at the time of placental formation. METHODS: Placental tissues from healthy cows (n = 3) in early pregnancy (3-5 months) were collected at the slaughterhouse. Maternal and fetal parts were manually divided prior to homogenization. Further analysis was done in triplicates on the maternal and fetal parts separately and subjected to one-dimensional (1D) electrophoretic separation, followed by identification of peptide maps by nanospray liquid chromatography/tandem mass spectrometry (nanoLC/MS/MS). Proteins were identified by use of the MASCOT software with the SwissProt database. RESULTS: Proteomic analysis showed more than 4000 differentially expressed proteins in maternal and fetal parts of placenta. Each part expressed around 900 proteins, of which ca. 90 were common. The identified proteins were analyzed in accordance to molecular function and their participation in biological processes. CONCLUSIONS: The obtained results provide new insight into the knowledge about biochemical characteristics of placenta (new proteins) and serve for further studies on the possible markers of physiological/pathological pregnancy or function of placenta. Moreover, our data can be a good starting point for further studies on the processes underlying the attachment of placenta.

Total antioxidant capacity and protein peroxidation intensity in seminal plasma of infertile and fertile dogs.

The aim of this study was to evaluate the total antioxidant capacity and protein peroxidation intensity in seminal plasma of infertile and fertile dogs. The study was conducted on 10 infertile and 10 fertile dogs of various breeds. Infertility was defined as conception failure at least three matings with different bitches. Semen was collected by manual manipulation. The sperm concentration and motility parameters were evaluated using CASA Hamilton Thorne, Vers. IVOS 12.3. The morphology of spermatozoa and the percentage of live and dead sperm cells were assessed microscopically, total antioxidant capacity and the content of SH-groups in seminal plasma were determined spectrophotometrically, the contents of protein peroxidation markers in seminal plasma, bityrosine and formylokinurenine, were determined using spectrofluorimetric methods. Sperm concentration and total sperm count were significantly (p < 0.05) lower in infertile dogs than in fertile dogs (99.92 ± 3 0.05 × 106 /ml vs. 282.07 ± 48.27 × 106 /ml; 214.19 ± 114.74 × 106 vs. 747.57 ± 210.94 × 106 , respectively). The percentage of spermatozoa with normal morphology and the most determined motility parameters differed significantly (p < 0.05) between both groups. The mean values of total antioxidant capacity in the seminal plasma were significantly (p < 0.05) lower (19.95 ± 20.94 vs. 25.66 ± 23.18 µmol/g protein), whereas the mean contents of bityrosine and formylokinurenine in seminal plasma were significantly (p < 0.05) higher in infertile dogs than in fertile dogs (3.71 ± 4.83 µg/mg protein vs. 1.55 ± 2.00 µg/mg protein and 0.37 ± 0.45 µg/mg protein vs. 0.14 ± 0.08 µg/mg protein, respectively). In conclusion, the obtained results suggest that the poor semen quality and infertility in dogs could be associated with lowered total antioxidant capacity and increased protein peroxidation in seminal plasma as a consequence of oxidative stress.

Antioxidative and oxidative profiles in plasma and saliva of cows in different ages and hormonal status.

The aim of this study was to evaluate changes in the antioxidant status and oxidative stress parameters in plasma and saliva in order to investigate the physiological conditions of dairy cows. Blood and saliva were collected from clinically healthy female calves (n = 18), sexually mature, non-pregnant cows (n = 19), and pregnant dairy cows (n = 15). Spectrophotometric and spectroflurimetric analyses were carried out in the body fluids of these animals. The level of total antioxidant capacity (TAC) in plasma reached the lowest (p < 0.05) value in the group of sexually mature, non-pregnant cows (2.375 ± 0.500 µmol/g). A significant negative correlation (tau b = - 0.248, p < 0.05) was found between TAC values detected in plasma and saliva of examined animals. The highest (p < 0.05) mean values of thiol groups were detected in both plasma (0.007 ± 0.0015 mmol/g) and saliva (0.276 ± 0,116 mmol/g) of mature, non­pregnant cows. Conversely, the highest (p < 0.05) levels of formylokinurenine concentration were detected in saliva (11.535 ± 3.785 µg/mg) and plasma (0.133 ± 0.0237 µg/mg) of pregnant dairy cows. A significant positive correlation (tau b = 0.255, p < 0.05) was also found between the bityrosine content detected in plasma and saliva of the examined cows. In conclusion, although with regards to antioxidant/oxidative parameters saliva reflects the content of plasma only in part, however it shows age-related differences that can be used in the description of the physiological status of cows.

Preliminary results of the placental decorin profile in bovine pregnancy and parturition.

Decorin is a small leucine-rich proteoglycan which is involved in multiple biological functions mainly as a structural and signaling molecule. Due to its biological properties in connective tissue, decorin may participate in remodeling of ECM during attachment and detachment of placenta within the course of pregnancy and at parturition in cows. The aim of the study was to detect the presence of decorin protein in bovine placental tissues and to evaluate its profile during pregnancy and at parturition. Placental tissues from healthy pregnant cows (2-5 month) were collected in abattoir (n = 10), while parturient tissues were obtained during caesarian section at physiological term (n = 6). Maternal and fetal parts were separated manually and subjected to homogenization and to quantitative ELISA and verification by Western blotting with anti-decorin antibodies. ELISA test showed that the concentration of decorin during pregnancy was higher in the fetal part of placenta compared with the maternal part (p < 0.001). Similar pattern was noted regarding to maternal and fetal samples derived from parturient cows. Our preliminary results demonstrate that the concentration of decorin is gestation time-dependent in healthy bovine placenta. Possible confirmation of the involvement of decorin in early pregnancy attachment and detachment of the placenta during parturition requires further research.

Preliminary analysis of the protein profile in saliva during physiological term and preterm delivery.

From a clinical point of view, easily obtainable and useful markers of a particular pathological status are required for appropriate diagnosis and treatment. Analysis of the proteomic profile of saliva may allow for the selection of potential marker of preterm delivery in humans. Saliva samples were collected from 12 patients diagnosed with threatened preterm delivery and 10 controls with uncomplicated pregnancies at the same gestational age. Samples were analysed using 2D electrophoresis. Based on statistical analysis, spots of interest were selected and collected from gels. Subsequently, spots were decoloured and proteins were identified by mass spectrometry using the matrix assisted laser desorption ionization­time of flight technique. The results of identification were compared with the Swiss­Prot database. A total of 1,393 spots were detected in the present study with 59 significantly different between control and preterm samples. Increased intensity of staining of 32 spots was observed in the premature delivery group compared with control patients and 27 spots were stained more intensely in the control group compared with the premature delivery group. A total of nine spots, which were significantly different between examined samples were identified and three of them exhibited increased intensity of staining in premature delivery group compared with controls, including dedicator of cytokinesis protein 1, metallothionein­2, guanylyl cyclase­activating protein 1. The six remaining spots included, epithelial­stromal interaction protein 1, serum albumin, tyrosine­tRNA ligase, cytoplasmic, protein chibby homolog 3, leukemia inhibitory factor receptor and adenosylhomocysteinase 3, and exhibited increased intensity of staining in healthy controls compared with premature delivery group. Further studies with an increased number of patients and identification of the complete protein profile are required to confirm the results of the present study and applicability of saliva as a source of disease biomarkers.

[Right atrial and right ventricular thrombus in a patient with hepatic carcinoma - a case report]. / Skrzeplina w prawym predsionku i w prawej komorze. Opis przypadku.

A case of a 62-year-old male with suspected aortic dissection is presented. Echocardiography did not confirm the initial diagnosis and revealed the presence of thrombus in the inferior vena cava, right atrium and right ventricle. Computerised tomography, followed by histological examination of liver biopsy, revealed hepatic carcinoma. The patient received anticoagulation and unfortunately died 14 days later. Symptomatology of the right-heart thrombus and thrombo-embolic complications in patients with neoplastic disorders are discussed.