Effects of cortisol on pregnancy rate and corpus luteum function in heifers: an in vivo study

To determine whether glucocorticoids affect the function of the bovine corpus luteum (CL) during the estrous cycle and early pregnancy, we examined the effects of exogenous cortisol or reduced endogenous cortisol on the secretion of progesterone (P4) and on pregnancy rate. In preliminary experiments, doses of cortisol and metyrapone (an inhibitor of cortisol synthesis) were established (n=33). Cortisol in effective doses of 10 mg blocked tumor necrosis factor-induced prostaglandin F(2α) secretion as measured by its metabolite (PGFM) concentrations in the blood. Metyrapone in effective doses of 500 mg increased the P4 concentration. Thus, both reagents were then intravaginally applied in the chosen doses daily from Day 15 to 18 after estrus (Day 0) in noninseminated heifers (n=18) or after artificial insemination (n=36). Pregnancy was confirmed by transrectal ultrasonography between Days 28-30 after insemination. Plasma concentrations of P4 were lower in cortisol-treated heifers than in control heifers on Days 17 and 18 of the estrous cycle (P<0.05). However, the interestrus intervals were not different between control and cortisol-treated animals (P>0.05). Moreover, metyrapone increased P4 and prolonged the CL lifespan in comparison to control animals (P<0.05). Interestingly, in inseminated heifers, cortisol increased the pregnancy rate (75%) compared with control animals (58%), whereas metyrapone reduced the pregnancy rate to 16.7% (P<0.05). The overall results suggest that cortisol, depending on the physiological status of heifers (pregnant vs. nonpregnant), modulates CL function by influencing P4 secretion. Cortisol may have a positive influence on CL function during early pregnancy, leading to support of embryo implantation and resulting in higher rates of pregnancy in heifers.     

Effects of the commercial extract of aronia on oxidative stress in blood platelets isolated from breast cancer patients after the surgery and various phases of the chemotherapy

Since the extract from berries of Aronia melanocarpa presents antioxidative properties in plasma and in blood platelets, not only from healthy group, but also from patients with benign breast diseases and in patients with invasive breast cancer before surgery, the aim of our present study was to evaluate the oxidative stress by measuring the level of various biomarkers of this process such as the generation of superoxide anion radicals (O(2)(-·)), the amount of carbonyl groups and 3-nitrotyrosine in proteins or the amount of glutathione in blood platelets isolated from breast cancer patients after the surgery and after various phases of the chemotherapy in the presence of A. melanocarpa extract (Aronox) in vitro. We demonstrated in platelet proteins from patients with invasive breast cancer (after the surgery and after various phases of the chemotherapy) higher level of carbonyl groups than in control healthy group. The level of 3-nitrotyrosine in platelet proteins from patients with invasive breast cancer was also significantly higher than in healthy subject group. We observed an increase of other biomarkers of oxidative stress such as O(2)(-·) and a decrease of GSH in platelets from patients with breast cancer (after the surgery and after various phases of the chemotherapy) compared to the healthy group. In model system in vitro our results showed that the commercial extract from berries of A. melanocarpa due to antioxidant action, significantly reduced the oxidative/nitrative stress in platelets from patients with invasive breast cancer caused by the surgery and various phases of the chemotherapy.     

Ectomycorrhiza formation and willow growth promotion as affected by associated bacteria: role of microbial metabolites and use of C sources

Biotic interactions affect the impact of potential plant growth promoting microorganisms in the rhizosphere, but their magnitude and fundamentals are often hardly known. In a pot experiment, two physiologically different strains of the ectomycorrhizal fungus Paxillus involutus (GUL and FRA) were tested separately and in combination with associated bacteria (Sphingomonas paucimobilis 1 L, Ralstonia pickettii 16 B, Sphingomonas sp. 23 L) on their effects on the growth of willows (Salix viminalis). Both P. involutus strains significantly increased the growth of the willows compared with non-inoculated plants, but the magnitude of this effect was significantly affected by the fungal strain. The P. involutus strain GUL with higher synthesis of auxin-like substances, acid phosphatases, siderophores and faster utilisation of yeast extract in vitro increased the willow growth in situ more effectively than strain FRA. Additionally, dual inoculation with the P. involutus strains GUL and FRA in combination with the associated bacteria promoted the willow growth, especially the combination with S. paucimobilis 1 L. This bacterial strain used effectively C sources which are common components of plant root exudates, e.g. glucose, sucrose, maltose and mannose as well as compounds synthesized by fungi, e.g. trehalose in vitro. We conclude that the analyses of fungal metabolites and of C source use of associated bacteria can successfully contribute to accelerate the selection of capable plant growth promoting combinations.     

Ecophysiological aspects of in vitro biotechnological studies using somatic embryogenesis of callus tissue toward protecting forest ecosystems

This review on current biotechnological methods in forestry for in vitro tissue cultures to define the effect of stress conditions on trees,concentrates on somatic embryogenesis.Callus tissue,the key product of somatic embryogenesis,grows over a tree wound under ex vitro conditions.Callus tissue can be used in research in areas such as pathogenic susceptibility at the embryonic level,effect of heavy metals,influence of low temperatures(cryopreservation),production of secondary metabolites and transformation of plants.Callus of arborescent plants can be induced in vitro by fungal elicitors to produce secondary metabolites for pharmaceutical and cosmetic industries and are strongly repellant to herbivores and can thus act to protect forests.Analyses of dual cultures demonstrated that callus tissue exposed to a pathogenic fungus responds by synthesizing low-molecular-mass proteins belonging to an immune protein class.Cryopreservation of embryonic callus tissue also has broad applications,e.g.,for valuable plant genotypes in gene banks.Without strategies to protect forests against stressfactors,forest ecosystems will degrade to the detriment of all life,including humans.In vitro biotechnological research using callus tissue contributes to progress in forestry and the disciplines of ecology,physiology,phytopathology,culture and selection of plants.     

Immunolocalisation of oestrogen receptors alpha (ERalpha) and beta (ERbeta) in porcine embryos and fetuses at different stages of gestation

The sites of oestrogen action can be shown by the localisation of their receptors in the target tissues. The aim of the present study was to show the localisation of oestrogen receptors in porcine embryos and fetuses obtained on days 18, 22, 32, 40, 50, 60, 71 and 90 post coitum (p.c.). The visualisation of proteins was conducted in embryos and various fetal organs such as gonads, uterus, lung, kidney, intestine and adrenal gland. Both ERs were observed in the blastocysts on day 18 p.c. In the male, ERbeta was detected in the testis and epididymis, whereas ERalpha was present in the efferent ductules. In the female, ERbeta was detected in the ovarian stromal cells investing the oocyte nests, while ERalpha protein was detected in the surface epithelium. In the uterus, ERs were present in the stromal cells, while ERbeta was present in the luminal epithelium. In the non-reproductive fetal porcine tissues ERbeta was localised in the lungs, kidneys, adrenal glands and in the umbilical cords. Both ERs were observed in the intestine. It is possible that ERbeta may play important roles in the development of the adrenal gland, testis, kidney and lungs, while both ERs are involved in the development of the ovary, uterus, epididymis and intestine of the porcine fetus.     

The effect of copper nanoparticles and copper (II) salt on redox reactions and epigenetic changes in a rat model

The aim of the study was to evaluate the effects of a diet containing different levels of Cu in two different chemical forms (carbonate and nanoparticles) on redox reactions and epigenetic changes in a rat model. For 4 weeks, five experimental groups (eight rats in each) were fed diets with two dosages of added Cu (standard—6.5 mg/kg or half of the standard dosage—3.25 mg/kg, and as a negative control no additional Cu in the mineral mixture) in two forms (standard—CuCO₃ and copper nanoparticles). Addition of Cu nanoparticles resulted in higher Cp (ceruloplasmin) activity and LOOH (lipid peroxides) and MDA (malondialdehyde) content, as well as decrease the CAT (catalase) activity and level of PC (protein carbonyl), 3‐NT (3‐nitrotyrosine), 8‐OHdG (8‐hydroxydeoxyguanosine), GSH + GSSG (total glutathione) and DNA methylation. Reducing the dose of copper resulted in a decrease in the level of LOOH and GSH + GSSG as well as CAT activity, but increased the level of PC and methylated DNA. Based on these evidence, we concluded that addition of copper nanoparticles in the diet reduces protein oxidation and nitration as well as DNA oxidation and methylation. Lowering the level of Cu in the diet increases the oxidation of proteins and DNA methylation.     

Occurrence of reproductive disorders in pig herds with and without Chlamydia suis infection – statistical analysis of breeding parameters

Chlamydiae are frequently encountered Gram‐negative intracellular eubacteria that can cause clear manifestations or clinically asymptomatic disorders. C. suis and other chlamydia are primarily isolated in cases of reproductive disorders. This study was performed to estimate the impact of Chlamydia suis infection on reproduction in sows by analyzing reproduction rates and breeding parameters. The test was conducted on first generation (F1) pigs from Polish Landrace (PL) × Polish Large White (PLW). Sixty‐four herds were investigated and 500 vaginal swabs were collected. Isolation of DNA was carried out directly from the swabs. All samples were analyzed for Chlamydia suis by real‐time PCR with a locked nucleic acid (LNA)‐containing probe. To analyze the impact of chlamydia infection on reproductive parameters, evaluation questionnaires were used. Reproductive problems were found in 77.3% of the farms tested. The receiver operating characteristic curve indicated that in the farms with 10 up to 120 sows, there were higher reproductive problems with chlamydia infection than in smaller and bigger pig farms. The most common problems were estrus repetition, which was reported by 57.81% of the surveyed farms, and the birth of dead piglets, which was reported by 31.25% of the investigated pig farms. Abortions, which were reported by 28.12% of the surveyed farms, were the least common reproductive disorders.     

Transformation of beech forest litter as a factor that triggers arsenic solubility in soils developed on historical mine dumps

Purpose

Soils that develop on the dumps in historical arsenic mining sites contain high concentrations of As thus constituting a serious environmental risk. This study was aimed to examine the changes in arsenic solubility in mine soils as induced by organic matter introduced with forest litter.

Materials and methods

Four large samples of initially developed soils were collected from the dumps remaining in former mining sites and were incubated for 90 days at various moistures: 80% of maximum water holding capacity and 100% (flooded conditions), with and without addition of beech forest litter (BL), 50 g/kg. Soils contained up to 5.0% As. Soil pore water was collected periodically with MacroRhizon suction samplers and examined on As, Mn, and Fe concentrations, pH, Eh, and dissolved organic carbon (DOC). The properties of dissolved organic matter were characterized by UV-VIS spectroscopic parameters A4/A6 and SUVA₂₅₄.

Results and discussion

Application of BL resulted in an intensive release of As from soils, particularly at 100% moisture. As concentrations in soil pore water increased strongly during the first 2 or 4 weeks of incubation and then started to decrease in all cases, except for one flooded soil. As was released particularly intensively from carbonate-containing soils. The mechanisms of As mobilization, including reductive dissolution of Mn and Fe oxides and the competition with DOC for sorption sites on the oxides, were discussed as related to soil properties. Pore water concentrations of DOC were increasing at the beginning of incubation and started to decrease after two or four weeks. Spectroscopic parameters of dissolved organic matter in ZS soils indicated increasing aromaticity and progress of humification.

Conclusions

Forest litter introduced to mine dump soils causes a mobilization of As into soil pore water. This effect, particularly strong in carbonate-rich soils, is apparently related to high concentrations of DOC and usually declines with time, which may be explained by the progress in humification. The relationships between DOC properties and As speciation in soil pore water should be dissected for better interpretation of experimental results.

     

The effects of Roundup on gametes and early development of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L)

To determine the effects of Roundup, a commercial formulation of glyphosate, gametes, and embryos of common carp (Cyprinus carpio L) was exposed to wide range of herbicide concentrations (0.0, 0.1, 0.5, 2.0, 5.0, 10.0, 20.0, and 50.0 mg/l). The obtained results showed different effects of Roundup on common carp gametes. Herbicide reduced swelling of eggs (but the effect was not concentration-related), while sperm showed low sensitivity to Roundup (time of spermatozoa motility was reduced in a significant way only at 20 mg/l, and at remaining concentrations, only a slight tendency was observed). During the embryonic development, Roundup caused a decrease of common carp embryonic survival (and the effect was concentration-related); however, it had no effect on development rate. During the embryogenesis, three types of embryo body malformation were observed: yolk sac edema, spine curvature, and shortening of body, but their frequencies were not associated with the presence or concentration of herbicide. However, Roundup affected quality of newly hatched larvae of common carp by increasing their mortality. No effect of herbicide on percentage of deformed larvae was observed but larvae hatched in water with Roundup tended to show more complex anomalies compared to those from the control. Obtained data showed that even low concentrations of this herbicide in waters can significantly reduce egg swelling, survival of embryos, and quality of fish larvae.