The influence of progesterone and prostaglandin F2α on decorin and the adhesion of caruncular epithelial cells of bovine placenta at early-mid pregnancy—Part II

One of the most important processes determining the proper course of gestation and its physiological termination in cows is the adhesion of epithelial cells allowing for direct contact of maternal and foetal parts of the placenta. Throughout pregnancy, placental cells are under strict hormonal control, which among others regulates the concentration and activity of specific proteins participating in the extracellular matrix remodelling of foetal membranes. The aim of the study was to evaluate the influence of progesterone and prostaglandin F_2α on the adhesion of epithelial cells at early-mid pregnancy in cows. Additionally, the impact of selected hormones on anti-adhesive properties of decorin was evaluated. Caruncular epithelial cells were isolated from healthy cows during pregnancy, immediately after slaughter. Primary cell cultures derived from the 2nd and 4th month of gestation were used in the experiments. The viability of cells was assessed by MTT assay. The adhesion of cells to fibronectin was measured spectrophotometrically. The activity of metalloproteinases was confirmed by the metalloproteinase assay. Progesterone (10^–5 and 10^–7 mol/L) and prostaglandin F_2α (10^–4_, 10^–5 and 10^–7 mol/L) increased the viability of bovine caruncular epithelial cells in the 2nd month of pregnancy. The treatment with prostaglandin F_2α significantly reduced the number of adherent cells from the 2nd month of gestation at the doses of 10^–4 and 10^–5 mol/L. Both progesterone and prostaglandin F_2α were shown to have an effect of decorin resulting in both a decrease in metalloproteinase activity and an increase in adhesion of cells to fibronectin.     

Proteomic profiling of plasma-derived small extracellular vesicles: a novel tool for understanding the systemic effects of tick burden in cattle

Cattle ticks pose a significant threat to the health and profitability of cattle herds globally. The investigation of factors leading to natural tick resistance in cattle is directed toward targeted breeding strategies that may combat cattle tick infestation on the genetic level. Exosomes (EXs), small extracellular vesicles (EVs) of 50 to 150 nm diameter, are released from all cell types into biofluids such as blood plasma and milk, have been successfully used in diagnostic and prognostic studies in humans, and can provide essential information regarding the overall health state of animals. Mass spectrometry (MS) is a highly sensitive proteomics application that can be used to identify proteins in a complex mixture and is particularly useful for biomarker development. In this proof of principle study, EXs were isolated from the blood plasma of cattle (Bos taurus) with high (HTR) and low tick resistance (LTR) (n = 3/group). Cattle were classified as HTR or LTR using a tick scoring system, and EXs isolated from the cattle blood plasma using an established protocol. EXs were subjected to MS analysis in data-dependent acquisition mode and protein search performed using Protein Pilot against the B. taurus proteome. A total of 490 unique proteins were identified across all samples. Of these, proteins present in all replicates from each group were selected for further analysis (HTR = 121; LTR = 130). Gene ontology analysis was performed using PANTHER GO online software tool. Proteins unique to HTR and LTR cattle were divided by protein class, of which 50% were associated with immunity/defense in the HTR group, whereas this protein class was not detected in EXs from LTR cattle. Similarly, unique proteins in HTR cattle were associated with B-cell activation, immunoglobins, immune response, and cellular iron ion homeostasis. In LTR cattle, unique exosomal proteins were associated with actin filament binding, purine nucleotide binding, plasma membrane protein complex, and carbohydrate derivative binding. This is the first study to demonstrate that MS analysis of EXs derived from the blood plasma of HTR and LTR cattle can be successfully applied to profile the systemic effects of tick burden.     

Diversity and abundance of springtails (Hexapoda: Collembola) in soil under 90-year potato monoculture in relation to crop rotation

The monoculture cropping system causes significant changes within the soil ecosystem, which constitutes a habitat for soil-dwelling springtails. Focusing on the response of soil fauna to 90 years of potato cultivation in monoculture the study investigates the abundance and diversity of soil-dwelling springtails, considering changes in the soil environment in relation to five-crop rotation. Another point was the soil quality evaluation using Collembola as bioindicators (QBS-c index). A long-term monoculture experiment was established in Poland in 1923 and has continued uninterruptedly to the present time. Soil samples were taken over a period of three years (2011–2013) to determine collembolan abundance and composition, as well as physical and chemical soil properties.

The study demonstrated that there were greater numbers of Collembola in the long-term monoculture of potatoes, especially before planting time, compared to numbers in a five-field crop rotation. At the same time apparently greater species diversity was found in potato culture within crop rotation. The biological indicator of soil quality based on the occurrence of springtails (QBS-c) has proved useful in assessing changes in soil caused by agrotechnical activities. This index indicated better biological soil quality in the five-field rotation system compared to monoculture.     

Protein recovery from rainbow trout (Oncorhynchus mykiss) processing byproducts via isoelectric solubilization/precipitation and its gelation properties as affected by functional additives

Solubility of rainbow trout proteins was determined between pH 1.5 and 13.0 and various ionic strengths (IS). Minimum solubility occurred at pH 5.5; however, when IS = 0.2, the minimum solubility shifted toward more acidic pH. Isoelectric solubilization/precipitation was applied to trout processing byproducts (fish meat left over on bones, head, skin, etc.), resulting in protein recovery yields (Kjeldahl, dry basis) between 77.7% and 89.0%, depending of the pH used for solubilization and precipitation. The recovered protein contained 1.4-2.1% ash (dry basis), while the trout processing byproducts (i.e., starting material) 13.9%. Typical boneless and skinless trout fillets contain 5.5% ash, and therefore, the isoelectric solubilization/precipitation effectively removed impurities such as bones, scales, skin, etc., from the trout processing byproducts. The recovered proteins retained gel-forming ability as assessed with dynamic rheology, torsion test, and texture profile analysis (TPA). However, the recovered proteins failed to gel unless beef plasma protein (BPP) was added. Even with BPP, the recovered protein showed some proteolysis between 40 and 55 degrees C. Addition of potato starch, transglutaminase, and phosphate to the recovered proteins resulted in good texture of trout gels as confirmed by torsion test and TPA. Higher ( P < 0.05) shear stress and strain were measured for gels developed from basic pH treatments than the acidic counterparts. However, proteins recovered from acidic treatments had higher ( P < 0.05) lipid content than the basic treatments. This is probably why the gels from acidic treatments were whiter ( L* - 3 b*) ( P < 0.05) than those from the basic ones. Our study demonstrates that functional proteins can be efficiently recovered from low-value fish processing byproducts using isoelectric solubilization/precipitation and subsequently be used in value-added human foods.     

Effect of Growth Regulators and Ethanol on Termination of Dormancy in Potato Tubers

The main objective of this study was to find the best practice of inducing the sprouting of dormant potato tubers. We compared two protocols of breakage of dormancy, which are based on dipping excised potato eyes in an aqueous solution of gibberellic acid (GA₃) and kinetin (standard 1) or in the aqueous solution of GA₃, thiourea, and daminozide (standard 2), with a newly reported approach based on ethanol. We tested the effect of ethanol alone or in combination with GA₃ and/or kinetin on dormancy release and sprouting of the potato tubers. As a model, we used two potato genotypes (cultivars Pasat and Dorota), with long dormancy of 5 and 10 weeks respectively. We showed that the standard 2 was the most effective treatment both for dormancy breaking and in promoting sprout growth, especially for cv. Dorota, for which the treatment induced 82.3% of tuber eye-plugs to sprout 28 days after treatment and to produce 93.2% of emerged plants after subsequent 28 days of cultivation in the greenhouse. For this cultivar, similar efficacy was observed for the combination of 4% ethanol with GA₃ and kinetin. The same concentration of ethanol combined with GA₃ but without kinetin was the most efficient treatment for breaking dormancy of cultivar Pasat. However, the difference between the various treatment combinations was statistically insignificant. Ethanol alone or in combination with kinetin poorly induced breakage of dormancy, confirming the main role of GA₃ in artificial dormancy breaking. Thus our study showed that the standard 2 is the most effective approach for breakage of dormancy at least with long term-dormancy cultivars.     

Respiratory and hematological response of tench, Tinca tinca (L.) to a short-term cadmium exposure

The effects of 3 h exposure to 96hLC50 of cadmium (4.5 mg dm⁻³) on oxygen consumption rate, and hematological parameters (RBC, WBC, erythrocyte and leukocyte pattern) of juvenile tench were evaluated. Oxygen consumption significantly decreased beginning from 24 h postexposure, and remained reduced until the end of the experiment (96 h postexposure). RBC gradually increased, together with the percentage of juvenile cells in circulation. On the other hand, cadmium induced damage to the red cells – the share of cellular anomalies significantly increased with time postexposure. They included abnormal cell shape, vacuolization, swelling, chromatin disintegration in the nucleus, and nucleus indentation. The exposed fish showed a gradual and significant decrease in WBC without a shift of lymphocyte/neutrophil proportion. No significant changes in thrombocyte count occurred. The results show that short-term exposure to cadmium reduced fish energetic metabolism, and suppressed immune abilities. The symptoms gradually developed after the end of exposure, and no recovery took place until␣96 h.     

Response of a Juvenile Cyprinid,Lake Minnow <Emphasis Type="Italic">Eupallasella perenurus</Emphasis> (Pallas), to Different Diets

Three commercial starters (Carp Starter, Uni Starter and Perla Plus) and one non-commercial, with frozen Chironomidae larvae as a reference diet, were evaluated for the intensive rearing of juvenile lake minnow Eupallasella perenurus, a cyprinid fish that is critically endangered in Poland. The growth, condition, survival, body deformities, and chemical body composition were studied. The 90-day laboratory experiment was performed at 22 °C with fish that were initially 24.6 mm (mean total lenth (TL)) and 0.11 g (mean body weight (BW)). Satisfactory fish growth was attained with all of the diets; however, the largest (p ≤ 0.05) final size (48.5 mm TL, 1.55 g BW) and the lowest condition coefficient (K = 1.34) were noted in fish fed the non-commercial starter. The final survival rates were very high (97.5–100%). Skeletal deformities (in 74 to 92% fish) were recorded exclusively in fish fed commercial starters. All commercial starters resulted in considerably higher lipid content and lower ash content than did the non-commercial starter and the reference diet. This suggests that both these factors might be responsible for body deformities. The present results proved that only the non-commercial starter is suitable for juvenile E. perenurus rearing under controlled condition, and that none of the commercial starters can be recommended.     

Does reduced MHC diversity decrease viability of vertebrate populations?

Loss of genetic variation may render populations more vulnerable to pathogens due to inbreeding depression and depletion of variation in genes responsible for immunity against parasites. Here we review the evidence for the significance of variation in genes of the Major Histocompatibility Complex (MHC) for conservation efforts. MHC molecules present pathogen-derived antigens to the effector cells of the immune system and thus trigger the adaptive immune response. Some MHC genes are the most variable functional genes in the vertebrate genome. Their variation is clearly of adaptive significance and there is considerable evidence that its maintenance is mainly due to balancing selection imposed by pathogens. However, while the evidence for selection shaping MHC variation on the historical timescale is compelling, a correlation between levels of MHC variation and variation at neutral loci is often observed, indicating that on a shorter timescale drift also substantially affects MHC, leading to depletion of MHC diversity. The evidence that the loss of MHC variation negatively affects population survival is so far equivocal and difficult to separate from effects of general inbreeding. Some species with depleted MHC variation seem to be particularly susceptible to infection, but other species thrive and expand following severe bottlenecks that have drastically limited their MHC variation. However, while the latter demonstrate that MHC variation is not always critical for population survival, these species may in fact represent rare examples of survival despite of the loss of MHC variation. There is clearly a compelling need for data that would disclose the possible consequences of MHC diversity for population viability. In particular, we need more data on the impact of MHC allelic richness on the abundance of parasites or prevalence of disease in populations, while controlling for the role of general inbreeding. Before such evidence accumulates, captive breeding programs and other conservation measures aimed at inbreeding avoidance should be favoured over those protecting only MHC variation, especially since inbreeding avoidance programs would usually conserve both types of genetic diversity simultaneously.     

Physiological and biochemical characterization of quinclorac resistance in a false cleavers (Galium spurium L.) biotype

The physiological and biochemical basis for quinclorac resistance in a false cleavers (Galium spurium L.) biotype was investigated. There was no difference between herbicide resistant (R) and susceptible (S) false cleavers biotypes in response to 2,4-D, clopyralid, glyphosate, glufosinate-ammonium, or bentazon. On the basis of GR(50) (growth reduction of 50%) or LD(50) (lethal dose to 50% of tested plants) values, the R biotype was highly resistant to the acetolactate synthase (ALS) inhibitor, thifensulfuron-methyl (GR(50) resistance ratio R/S = 57), and quinolinecarboxylic acids (quinclorac R/S = 46), resistant to MCPA (R/S = 12), and moderately resistant to the auxinic herbicides picloram (R/S = 3), dicamba (R/S = 3), fluroxypyr (R/S = 3), and triclopyr (R/S = 2). The mechanism of quinclorac resistance was not due to differences in [(14)C]quinclorac absorption, translocation, root exudation, or metabolism. Seventy-two hours after root application of quinclorac, ethylene increased ca. 3-fold in S but not R plants when compared to controls, while ABA increased ca. 14-fold in S as opposed to ca. 3-fold in R plants suggesting an alteration in the auxin signal transduction pathway, or altered target site causes resistance in false cleavers. The R false cleavers biotype may be an excellent model system to further examine the auxin signal transduction pathway and the mechanism of quinclorac and auxinic herbicide action.     

Influence of Agricultural Land Use and Management on the Contents of Polycyclic Aromatic Hydrocarbons in Selected Silty Soils

The characteristics of transport and transformation of SO₂, NO_x and O₃ in northeast Asia have been investigated by using a comprehensive regional air quality model (RAQM) driven by a meteorological model MM5. A study period of 1-15 March 2002 has been selected due to the availability of intense observation of chemical species for both ground and upper levels. Model results have been compared against observational data to provide insights into the strength and weakness of the model’s ability and the evolutionary features of chemical species. Validation shows a good skill of this model system in reproducing most of the key features in long-range transport, but apparent bias still remains due to a series of uncertainties from either emission estimates, prescribed parameters, or inherent model limitations. In general, this model shows a better skill for SO₂ and O₃ than for NO_x. Large discrepancy occurs between the observed and calculated NO_x concentration at higher levels, with the model results being much lower. A series of sensitivity tests have been conducted to investigate the potential affecting factors and it is found that the inaccuracy or incompleteness in currently used emission inventories could be a most likely cause for such discrepancy. Long-range transport from Asian continent to the western Pacific is pronounced in springtime. Substantially high concentrations of SO₂ and NO_x in or above the boundary layer (0.5～3.0 km) over the Yellow Sea suggest an important pathway for long-range transport in northeast Asia.