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The objective of this research was to assess the viability and developmental potential of feline oocytes following in vitro maturation (IVM), vitrification, and post-warming incubation with resveratrol. In the first experiment, warmed oocytes were incubated with 0.2 μM, 2 μM, or 20 μM resveratrol for 2 h. Oocytes treated with 0.2 μM resveratrol had the highest viability (68.89 %), as assessed by fluorescein diacetate and ethidium bromide staining, while higher concentrations were associated with diminished oocyte viability. In the second experiment, the warmed oocytes were inseminated following the 2-h incubation with the three concentrations of resveratrol. The presumptive zygotes were then maintained in culture and their development evaluated. The highest cleavage rate was observed when the oocytes had been incubated with 0.2 μM resveratrol (88.34 %), which was higher than for the control group (without resveratrol (75 %)). Moreover, this concentration of resveratrol also augmented the blastocyst formation rate. While the vitrification of oocytes often results in diminished developmental potential in the ensuing embryos, attributed to cryopreservation-induced injury, the utilization of low concentrations of resveratrol enhances the procedure's efficacy. • Resveratrol treatment is a possible countermeasure against cryopreservation‐induced mitochondrial damage. • Resveratrol improved the viability of oocytes after vitrification. • Oocytes vitrified and incubated with resveratrol have good developmental competence after IVF. • Resveratrol improves the quality of oocytes vitrified in minimal volumes. [ABSTRACT FROM AUTHOR]

Bipolar radiofrequency ablation (B-RFA) is a method used to treat the arrhythmia substrate resistant to unipolar ablation. Few studies have addressed endo-epicardial B-RFA. The aim of the study was to evaluate chronic lesions resulting from endo-epicardial B-RFA and to determine optimal settings for such procedures in an animal model. In 7 pigs, up to 5 radiofrequency applications per animal were performed with 2 electrodes placed on both sides of the left ventricular free wall. Current was delivered for 60 seconds by a generator dedicated for B-RFA with power settings of 25, 30, 35, 40, and 50 W. At 12 weeks after ablation, 31 lesions were assessed. Their maximal cross-sectional area ranged from 7.2 to 68 mm2 and correlated with total power delivered (r = 0.53), with temperature increment at the endocardial catheter (r = 0.65), and inversely with temperature decrement at the epicardial catheter (r = 0.54). For power values between 30 and 40 W, the lesion area did not differ significantly (P =.92). Lesion depth ranged from 1.9 to 11 mm and correlated with impedance decrement (r = 0.5). Lesions were transmural in 8 cases. Lesion depth/wall thickness ratio was on average 0.6 ± 0.3, with the smallest value for 25 W (0.5 ± 0.3) and the largest for 50 W (0.8 ± 0.3). Steam pops occurred at a power range of 30–50 W, with an incidence of 1 in 5 applications, with 1 case of fatal tamponade at 40 W. Impedance decrement, endocardial catheter temperature increment, and endocardial electrogram amplitude decrement were greater during applications with steam pops. Chronic lesions resulting from endo-epicardial B-RFA appear smaller and less often transmural compared with acute lesions described in the literature. The incidence of steam pops during endo-epicardial B-RFA is relatively high even at low powers. [ABSTRACT FROM AUTHOR]

The present experiment employed time-lapse (TL) imaging to assess the effects of vitrification on the development of ovine blastocysts and to see if the timing of blastocyst formation and expansion was correlated with the numbers of embryo- and trophoblasts determined through differential staining of the expanded blastocysts. Ovaries were obtained after slaughter from cycling (October–March) Polish Longwool ewes aged 1–3 years and cumulus-oocyte complexes were collected by ovarian scarification. In vitro maturation was performed in TCM 199 medium supplemented with Earle's Salt, 10% of FBS, and 5 μg/mL of LH/FSH at 38 °C for 24 h. After maturation, the oocytes were incubated with thawed ram semen (IVF) for 19 h and all presumptive zygotes were transferred to a 16-well dish containing Cult medium for monitoring with the Primo Vision TL system. A portion of ovine embryos were vitrified (Cryotop system) at the early cavitation stage and TL observations of warmed (n = 30) and non-vitrified (n = 32) blastocysts continued until the attainment of the expanded blastocyst stage, at which point they were differentially stained with bisbenzimide and propidium iodide for microscopic enumeration of embryoblasts (inner cell mass blastomeres) and trophoblast-cells. There were no statistically significant differences in the timing of blastulation (tB) and formation of expanded blastocysts (tEB) between vitrified and non-vitrified ovine embryos, but non-vitrified blastocysts exceeded (P < 0.05) their vitrified and warmed counterparts in the mean number of embryo- and trophoblasts. In addition, the number of trophoblasts was negatively and moderately correlated with tB and tEB, for both vitrified and non-vitrified embryos. It can be concluded that even though vitrification of ovine embryos is associated with a significant reduction in the number of blastomeres, the rate of blastocyst development remains closely linked to the numbers of trophoblastic cells. • Vitrification reduced the number of trophoblasts cells in warmed ovine embryos. • Vitrification of ovine embryos is associated with a significant reduction in the number of blastomere. • There is relationship between the timing of blastocyst formation and expansion and the number of trophoblasts. • Embryonic morphokinetics are determinants and predictors of in vitro fertilization outcomes. • The rate of trophoblast expansion is a significant potential marker of embryo quality. [ABSTRACT FROM AUTHOR]

The aim of this study was to compare several culture systems for cat embryos. Domestic cat oocytes were matured in vitro (IVM), fertilized (IVF), and cultured individually or in groups in drops under oil (20 μL or 50 μL) and in 16 microwell dishes (Primo Vision®). Moreover, the effects of co-culture with a) uncleaved oocytes, b) homospecific and c) heterospecific co-culture with cat and sheep companion embryos were investigated using a time-lapse system. A higher proportion of blastocysts and hatching blastocysts was observed after culture in Primo Vision® dishes compared with the classical individual (p < 0.001) and group (p < 0.05) culture systems. Culture of presumptive zygotes 16 hpi and the presence of uncleaved oocytes did not reduce blastocyst development compared with culture of embryos 24 hpi without uncleaved oocytes. Co-culture with later-stage companion cator sheep embryos accelerated development of catembryos. The highest percentage of blastocysts was obtained in the group co-cultured with sheep embryos (54%). Moreover, the blastocyst cavity formed on average 10 h faster in this group than for the control group and for embryos co-cultured with cat embryos. The proportion of hatching blastocysts was similar in the co-cultures with cat and with sheep embryos (20% vs. 22%) and significantly (p < 0.05) than in the control group (12%). • Effect of embryo culture system (individual, group, WOW) on the development potential of the cat embryos. • Effect of co-culture with non-cleaved oocytes on the development of feline embryos. • Co-culture of feline embryos with heterospecific companion embryos. [ABSTRACT FROM AUTHOR]

Time-lapse (TL) imaging provides a practical and safe tool to constantly monitor the development of in vitro -derived embryos. TL may help develop novel methods of predicting the timing of embryo cleavage that will lead to optimizing blastocyst cryopreservation or transfer. The primary objective of the present study was to employ TL imaging to examine associations among the division kinetics of ovine embryos, their quality and rates of development to the blastocyst stage. Oocytes were collected by ovary scarification from 78 Longwool ewes slaughtered in the breeding season (November–March). Cumulus oocyte complexes (COCs) were matured for 24 h in TCM 199 media containing 0.1 IU/mL LH/FSH and 10% FBS. In-vitro fertilization was carried out by co-incubation of semen and COCs for 19 h. Presumptive zygotes were placed in microwells, in droplets of Cult medium (Gynemed, Lensahn, Germany). Digital images of developing embryos were captured every 10 min by Primo Vision TL system (EVO+; Vitrolife, Göteburg, Sweden). The following time intervals were recorded: from IVF to the attainment of two-cell (t2), three-cells (t3) or four-cell (t4) stage, to morula detection (tM), blastulation (tSB) and blastocyst formation (tB). Lastly, the duration of the second cell cycle (cc2; t3-t2) and complete synchronous cell division (s2; t4-t3) were calculated, and the incidence of developmental anomalies noted. Out of 147 embryos selected for TL observations, 55 (37.4%) developed to the blastocyst stage (normally developing embryos, NE) and 92 (62.6%) failed to reach the blastocyst stage (arrested embryos, AE; P < 0.05). Mean t2, tM, s2 and cc2 were all less (P ≤ 0.02) in NE compared with AE. Approximately 61.9% of embryos exhibited developmental anomalies (35.5% in the NE group and 78.2% in the AE group; P < 0.05) and AE exceeded (P < 0.05) NE in the proportion of FRG (blastomeric fragmentation), IRR (blastomeres of irregular size after cleavage), DC (direct cleavage) and MA (multi-morphological aberrations). Of all NE, 63.6% were classified as good quality and 36.4% as poor quality blastocysts (P < 0.05). Good quality ovine blastocysts attained t2, t3, t4, tSB and tB stages earlier (P ≤ 0.03) than poor quality blastocysts and none of the poor quality blastocysts was seen to hatch. To recapitulate, the present results indicate that the kinetics of early ovine embryo development are significant predictors of their potential to develop to the blastocyst stage and the markers of blastocyst quality. Time-lapse imaging may serve as a useful technique for predicting the outcome and enhancing efficacy of in vitro embryo production in sheep. • Time-lapse imaging permits continuous, non-invasive monitoring of sheep embryos. • Time of early embryo cleavage affects the quality of ovine blastocysts in vitro. • Morphological aberrations occur more frequently in arresting sheep embryos. • Good quality ovine blastocysts reach blastocyst stage earlier. • Morphokinetic parameters are associated with ovine embryo development. [ABSTRACT FROM AUTHOR]

[Display omitted] • Biopreparation may be used in the degradation of drugs in real conditions. • Bioaugmentation does not disturb the functioning of activated sludge. • The carrier's long-term presence in the activated sludge leads to its decomposition. The presence of non-steroidal anti-inflammatory drugs in wastewater from sewage treatment plants indicates that they are not completely biodegradable. The designed biopreparation based on immobilized bacteria enables the degradation of paracetamol, ibuprofen, naproxen and diclofenac at a rate of 0.50 mg/L*day, 0.14 mg/L*day, 0.16 mg/L*day and 0.04 mg/L*day, respectively. Lower degradation of drugs in the mixture than in monosubstrate systems indicates their additive, antagonistic effect, limiting the degradative capacity of microorganisms. The biopreparation is stable for at least 6 weeks in bioreactor conditions. Biochemical parameters of activated sludge functioning showed increased oxygen demand, which was related to increased ammonia concentration caused by long-term exposure of activated sludge to drugs. Reduced metabolic activity was also observed. The preparation enables decomposing drugs and their metabolites, restoring the activated sludge's functionality. The tested biopreparation can support activated sludge in sewage treatment plants in degrading non-steroidal anti-inflammatory drugs and phenolic compounds. [ABSTRACT FROM AUTHOR]

Abstract This study aimed to characterize the landscape sustainability in terms of landscape services (LS) provided in seven study areas with different characteristics located in Malopolska Province. The results of the qualitative categorization of LS using an assessment matrix were juxtaposed with the results of the quantitative assessment using indicators. As a result, study areas were divided into three groups regarding the bundle of provided LS. In multifunctional landscapes characterized by moderate flow of most LS and often high flow of cultural LS, there were: high landscape diversity, moderate connectivity of forest patches and moderate values of regulating LS. Mountainous landscape with a high flow of regulating LS was characterized by moderate landscape diversity, high connectivity of forest class and high values of erosion control and climate regulation. In agricultural landscapes with a high flow of agricultural production LS, there were: low landscape diversity and connectivity of forest patches, low values of erosion control and climate regulation, but high values of nutrient retention. Consequently, all analyzed landscapes may be described as sustainable in terms of LS, as every landscape provides a specific bundle of LS and a reasonable level of landscape diversity, connectivity and regulating LS. The qualitative categorization of LS provided the most complete perspective on LS provided in the study areas. Landscape metrics (LM) allow the significance of landscape diversity and connectivity to be emphasized, whereas the quantitative analysis of regulating LS allow to compare the results of quantitative assessment and qualitative categorization. These two types of indicators are especially useful when threshold values are required. Composition metrics (proportion of forest and arable land) allow a determination of landscapes dominated by regulating and provisioning LS, whereas configuration metrics (especially Shannon’s Diversity Index and Contagion Index) allow landscapes with a high flow of cultural LS to be identified. [ABSTRACT FROM AUTHOR]

The rat nucleus incertus (NI) contains GABA/peptide-projection neurons responsive to orexin (hypocretin)/orexin receptor-2 (OX 2 ) signalling. Melanin-concentrating hormone (MCH) and orexin neurons often innervate and influence common target areas. Therefore, we assessed the relationship between these hypothalamic peptidergic systems and rat NI, by investigating the presence of an MCH innervation and MCH receptor-1 (MCH 1 ) expression, and neurophysiological and behavioural effects of MCH c.f. orexin-A (OXA), within the NI. We identified lateral hypothalamus (LH), perifornical and sub-zona incerta MCH neurons that innervate NI, and characterised the rostrocaudal distribution of MCH-containing fibres in NI. Single-cell RT-PCR detected MCH 1 and OX 2 mRNA in NI, and multiplex, fluorescent in situ hybridisation revealed distinct co-expression patterns of MCH 1 and OX 2 mRNA in NI neurons expressing vesicular GABA transporter (vGAT) mRNA. Patch-clamp recordings revealed 34% of NI neurons tested were hyperpolarised by MCH (1 μM), representing a distinct population from OXA-sensitive NI neurons (35%). Intra-NI OXA infusion (600 pmol) in satiated rats during the light/inactive phase produced increased locomotor activity and food (standard chow) intake, whereas intra-NI MCH infusion (600 pmol) produced only a trend for decreased locomotor activity and no effect on food intake. Furthermore, in satiated or pre-fasted rats tested during the dark/active phase, intra-NI infusion of MCH did not alter the elevated locomotor activity or higher food intake observed. However, quantification of neuropeptide-immunostaining revealed differential diurnal fluctuations in orexin and MCH trafficking to NI. Our findings identify MCH and orexin inputs onto divergent NI populations which may differentially influence arousal and motivated behaviours. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the effect of different concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on selected parameters of equine gametes under controlled in vitro conditions. Assessments covered maturation of oocytes as well as sperm-cell function and structure. In the first part of the experiment, the influence of different concentrations of TCDD (0.32 ng, 3.2 ng, and 32 ng/mL) on in vitro maturation (IVM) of equine oocytes was evaluated. During IVM, 28.38% of the oocytes reached the metaphase II (matured) stage at a TCDD concentration of 0.32 ng/mL and only 5.14% at a concentration of 3.2 ng/mL. In group, with the highest concentration of TCDD in the medium (32 ng), matured oocytes were not observed. In the second part of experiment, the influence of TCDD (0.32 ng, 3.2 ng, and 32 ng/mL) on sperm parameters, such as motility, plasma membrane integrity, acrosome integrity, mitochondrial activity, lipid peroxidation (LPO), apoptosis, and chromatin status, was evaluated. The present study demonstrates that moderate concentrations of TCDD may activate population of motile sperm cells under in vitro conditions with no influence on sperm-cell structure or LPO. Moreover, the modulation of motility function of sperm was confirmed by the detection of the evident increase of cell number with high mitochondrial potential. Dioxins exert negative impact on both female and male reproductive systems. Thus, an overall analysis of both systems will advance scientific knowledge on the impact of toxic agents on the reproductive system. [ABSTRACT FROM AUTHOR]

[Display omitted] • Various elements influence micro-climate around building and impact its energy use. • Micro-climatic impact on buildings in sub-tropical/tropical zones is rarely studied. • Passivhaus building performance in sub-tropical/tropical zones is under examined. • Passivhaus buildings are sensitive to surrounding micro-climates in temperate zones. • Digitised Eco-feedback is effective in increasing energy consumption awareness. The earth's climate differs enormously across its breadth with numerous small-scale variations to surrounding atmospheric conditions (micro-climates) existing side by side. These differences impact indoor environments of residential buildings and, depending on occupants' preferences, building energy use, and can mean that home designs are unfit for the local context. Although considered leaders in producing energy efficient designs which are equally comfortable for occupants and the environment, even Passivhaus (PH) homes have been demonstrated to be sensitive to localised outdoor conditions. Meanwhile, occupants have little understanding of their own domestic energy use, how the situation of their property might impact their comfort, and of more sustainable alternatives open to them. To boost understanding of household energy use and to encourage energy conservation, instantaneous computerised feedback on household's energy consumption (Eco-feedback) has been widely used. Along with improving household energy use understanding, Eco-feedback technologies may assist in comprehending local micro-climatic conditions around a building, and in learning how to manipulate these can help to establish and sustain preferred indoor conditions with minimal energy use. This can be achieved through producing personalised and contextualised feedback and responses to update and renovate users' homes by suggesting short-term actions and long-term design recommendations. This systematic literature review paper explores the breadth of Eco-feedback techniques, their relative benefits and shows how these could be deployed in the context of understanding building performance in the home. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the present study was to determine the vitality and developmental competence of equine oocytes after in vitro maturation (IVM) and vitrified by Rapid-i method. In experiment 1, oocytes after IVM were vitrified using media: EquiPro VitKit (group 1) or medium containing 18% Ficoll, 40% ethylene glycol, and 0.3 M sucrose (group 2). For evaluation of toxicity effect, oocytes were exposed to media without a plug to liquid nitrogen. To evaluate viability, oocytes were stained with fluorescein diacetate and ethidium bromide. In experiment 2, oocytes after IVM and vitrification were activated by 7.5 μM ionomicin in TCM 199 (5 minutes) combined with 2 mM 6-DMAP in TCM 199 with 10% fetal bovine serum (4.5 hours). Survival rate was: 63% in group 1 (n = 54), 55% in group 2 (n = 69), and 73.2% (n = 56) in the control group. After parthenogenetic activation, 10.2% (n = 49) of 2–4 blastomeres were observed. This percentage was lower than in the nonvitrified group: 38.5% (n = 53). [Copyright &y& Elsevier]

Abstract: Activities of 19 hydrolases were estimated in cell suspensions of 40 Brochothrix thermosphacta strains, isolated from meat and meat products packaged under various conditions, by using API ZYM® test. These strains produced: acidic phosphatase, esterase C4, esterase/lipase C8, α-chymotrypsin, leucine arylamidase, β-glucosidase and α-glucosidase. The predominating biotype produced only the first four of the listed enzymes. The extracellular lipolytic activity for bromo-chloro-indolyl-caprylate was detected only in culture broth supernatants of 4 strains thereby indicating that esterase/lipase C8 is cell-bound. 13 of the strains displayed proteolytic activity for albumin at 4°C (0.31–2.07U) while 9 strains showed this activity at 25°C (0.11–1.21U). Only 4 strains digested albumin at both temperatures. Thus the meat spoilage potential of B. thermosphacta strains results not only from digestion of carbohydrates but also from their proteolytic activity. [Copyright &y& Elsevier]

Drosophila Nurf55 is a component of different chromatin- modifying complexes, including the PRC2 (Polycomb repressive complex 2). Based on the 1.75-Å crystal structure of NurfS5 bound to histone H4 helix 1, we analyzed interactions of Nurf5S (NurfSS or p55 in fly and RbAp48/46 in human) with the N-ter- minal tail of histone H3, the first helix of histone H4, and an N-terminal fragment of the PRC2 subunit Su(z) 12 using isothermal calorimetry and pulldown experiments. Site-directed mutagenesis identified the binding site of histone H3 at the top of the Nurf55 WD4O propeller. Unmodified or K9me3- or K27me3-containing H3 peptides were bound with similar affinities, whereas the affinity for K4me3-containing H3 peptides was reduced. Helix 1 of histone H4 and Su(z)12 bound to the edge of the β-propeller using overlapping binding sites. Our results show similarities in the recognition of histone H4 and Su(z)12 and identify Nurf55 as a versatile interactor that simultaneously contacts multiple partners. [ABSTRACT FROM AUTHOR]

Abstract: Twenty Brochothrix thermosphacta strains tested for biogenic amines (BAs) production, formed histamine (6.6–16.2mg/kg) and tyramine (18.7–35.4mg/kg) but neither putrescine nor cadaverine. Six of the twenty strains were also investigated in respect of their influence on the synthesis of BAs by Escherichia coli, Pseudomonas sp., Proteus mirabilis and Lactobacillus sakei. In pure culture Escherichia coli produced all of the studied amines (histamine, tyramine, putrescine and cadaverine) with a total concentration of 167.7mg/kg, P. mirabilis produced a total of 56.7mg/kg histamine, tyramine and cadaverine, while Lactobacillus sakei and Pseudomonas sp. produced histamine and tyramine, totaling 37.9 and 35.2mg/kg respectively. All B. thermosphacta promoted cadaverine production by Escherichia coli which increased by 12–68%, and some of them contributed to the appearance of this amine among the metabolites of Pseudomonas. The presence of B. thermosphacta decreased the potential ability of P. mirabilis to produce BAs. [Copyright &y& Elsevier]

Abstract: We describe an automatic biodetector for continuous monitoring of water toxicity (ABTOW). Construction of the ABTOW is based on natural ability of the biofilm formation to immobilize consortia of nitrifying bacteria (the sensing element) on the open cellular polyurethane foam as the support. Change of rates of oxygen consumption is used as an indicator of biocatalytic activity (nitrification) of the bacteria in response to xenobiotics. Owing to this design the ABTOW features stability long-term use, is inexpensive and simple in operation. The dynamics of ABTOW response is studied in details for phenol and cyanide as model toxins. These data indicate that the sensitivity was 3.5μM for phenol and 0.19μM for cyanide, respectively. The magnitudes of toxic effect were proportional to concentration whereas kinetics of the response is an indicator for the mechanism of toxicity. Similar methodology is applied to quantify toxicity of a range of heavy metals, herbicides and oxidative chain inhibitors. One may conclude that the presented biodetector provides a good sensitivity for continuous on-line monitoring of toxicity in water. [ABSTRACT FROM AUTHOR]

Abstract: Automatic biodetector of water toxicity is a biosensor based on monitoring of catalytic activity of the nitrifying bacteria. To create a standardized biosensing system, development of the biofilm must be characterized to determine the prerequisites for its biological (biocatalytic) stability. In this paper, growth of biofilm comprising ammonium-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) in the open cellular polyurethane material polyurethane sponge bioreactor has been investigated. Dynamics of the biofilm formation was estimated using AOB and NOB metabolic activity and the volume occupied by these two types of bacteria in the biofilm. Spectrophotometry liquid ion chromatography and image cytometry were used, respectively, for these measurements. A mathematical model of the dynamics of biofilm formation was established. These data indicate that open cellular polyurethane material is a good basis for the immobilization of nitrifying bacteria. Moreover, growth of the biofilm leads to its stable structural form, whose biocatalytic activity (12.29 for AOB and 6.84μmolmin−1 for NOB) is constant in the long term. [Copyright &y& Elsevier]

Pseudomonas moorei KB4 belongs to the paracetamol-degrading strains. This strain was immobilized on a biodegradable carrier – a cellulose sponge from Luffa cylindrica. The study aimed to determine the impact of the immobilization process on the metabolic activity of the strain, including the biodegradation process of paracetamol. The research that was conducted showed significant differences in the level of transcription of selected genes, as well as in the metabolic profile and the composition of total fatty acids. However, the immobilization process did not significantly affect the degradation of paracetamol. This indicates a lack of limitation in the availability of the substrate, which is not very toxic to the tested strain. However, immobilization causes greater strain resistance to the appearance of the toxic 4-aminophenol. After immobilization, it is possible to decompose higher concentrations of paracetamol, which, under normal conditions, leads to the accumulation of 4-aminophenol, inhibiting the free strain's growth. Differences in the degradation of this drug by free and immobilized cells, depending on the number of doses, were observed. After analyzing the enzymes and intermediates of the paracetamol degradation pathway, differences were shown between the metabolism of this compound by the free and immobilized strain. It was shown that, 3-methyl-3-vinyl-cyclohexanon was a characteristic intermediate identified only during paracetamol degradation by the free strain of KB4. Moreover, no deaminase and hydroquinone 1,2-dioxygenase were active in the system with immobilized KB4. The resulting hydroquinone ring was probably not cleaved in the immobilized system. Hence, decomposition probably proceeds by catechol cleavage. • Immobilization affects the level of gene transcription. • Impoverishment of the metabolism of the KB4 strain after immobilization is observed. • No changes in the rate of paracetamol degradation after the immobilization. • Lack of hydroquinone cleavage by the immobilized KB4 strain. [ABSTRACT FROM AUTHOR]

• The leaf extracts decrease bacteria viability. • The leaf extracts act as prooxidants to increase ROS generation in bacteria cells. • The leaf extracts reduce bacteria membrane integrity. • The leaf extracts cause morphological changes in bacteria cells. • The leaf extracts inhibit DNA gyrase activity. The aim of this study was to investigate the in vitro antibacterial mechanisms of Aronia melanocarpa, Chaenomeles superba, and Cornus mas leaf extracts towards meat spoilage and pathogenic bacteria. The extracts decreased bacterial viability after 24 h and 48 h of incubation. Acting as prooxidants, the extracts induced intracellular ROS (reactive oxygen species) generation in bacteria cells, with C. mas having the strongest influence. The leaf extracts increased the release of UV intracellular absorbing components, suggesting a reduction in membrane integrity. They also increased the outer-membrane permeability of the Gram-negative bacteria, with C. superba extract being the most active. Following exposure to the leaf extracts, morphological changes in the bacteria were observed, including the formation of aggregates, EPS synthesis, irregular forms, wrinkled cell surfaces, pores in the cell wall, and shriveling of cells. The leaf extracts inhibited DNA synthesis in E. coli cells by suppressing DNA gyrase activity. [ABSTRACT FROM AUTHOR]

The in vitro maturation (IVM) of equine oocytes is still not efficient and does not yield consistent results. The specific requirements of equine oocytes during this process are still largely unknown, which hinders the development of assisted reproductive techniques (ART) in this species. Because the ovarian follicle microenvironment supports oocytes in their acquisition of developmental competence, follicular fluid seems to be a substantial source of bioactive factors that could support the IVM process. Extracellular vesicles (EVs) are cell-secreted molecules in body fluids that are able to deliver molecular signals and transfer genetic information (mRNA, miRNA) between donor and recipient cells. Hence, our hypothesis is that follicular fluid EVs (ffEVs) from small (<20 mm) ovarian follicles can improve the in vitro maturation rate of mare oocytes. To test our hypothesis, equine ovarian follicular fluid was aspirated and ffEVs were isolated by ultracentrifugation, then characterized using nanoparticle tracking analysis and flow cytometry. Additionally, ffEVs were labeled using the ExoGlow-protein EV labeling kit (System Biosciences, Palo Alto, CA). Cumulus-oocyte complexes (COCs) were matured using a one-step method (Method I, continuous culture for 24–38 h) or a two-step method (Method II, initial denudation after 24 h), in the presence (200 μg protein/ml) or absence of ffEVs. The results show the internalization of ffEVs by equine cumulus cells and, for the first time, also by oocytes. The ffEV treatment during two-step culture had a positive effect on the maturation rate of compacted COCs compared to the control group (45.7% and 20.5%, respectively; p < 0.05). No effect of supplementation was observed on the maturation rate during one-step culture. Our results indicate that the supplementation of culture media with EVs isolated from the follicular fluid of small follicles can improve the IVM rate of mare oocytes, suggesting that ffEVs play an important role during this process and may enhance the development of equine ART. • We observed the internalization of ffEVs by both equine cumulus cells and oocytes. • ffEV treatment during two-step culture had a positive effect on the maturation rate of compacted oocytes compared to the control group. • Supplementation of culture media with ffEVs from small follicles can improve the IVM rate of mare oocytes. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the study was to evaluate the influence of the Sildenafil citrate on the blood flow in the uterus of cows during dioestrus. Uterine blood flow was examined in five, healthy, adult cows. Between day 6–8 of the ovarian cycle, each cow received 200mg of sildenafil diluted in 10ml of warm saline into the body of the uterus. Analysis of the blood pressure, ECG and the maximum velocity in m/s (V max) in the aorta was performed and selected parameters of the blood flow (PI, pulsatile index; RI, resistance index; SPV, systolic peak velocity; EDV, end diastolic velocity; FVI, flow velocity integral; SV/DV, systolic peak velocity: end-diastolic velocity ratio) were measured in the uterine artery (Arteria uterine) before and after sildenafil infusion. In addition, Color Doppler examination of the uterine wall perfusion was analyzed. A significant decrease of values of PI and SV/DV ratio as well as an increase of end diastolic velocity and time averaged maximum velocity was noted. With the use of color coded sonography, the increased intensity of the blood flow in the uterine wall was observed. It was concluded that intrauterine administration of sildenafil during dioestrus can increase uterine tissue perfusion. [Copyright &y& Elsevier]

Abstract: The aim of this study was to verify the usefulness of the measurement of chosen cardiovascular activity parameters for examination of sex pheromones effects in male dogs. We evaluated the influence of the bitches’ sexual pheromones (BSP) on heart rate (HR) with the use of a Holter monitor, and chosen parameters of blood flow in vessels of penis, testes and kidney, with color-coded and pulsed Doppler ultrasonography. We found that the BSP increased HR in all examined males even without showing any other signs of arousal. There were no changes in the RI (resistant index) and PI (pulsative index) in any trials during the pheromone presentation. The increase in blood flow intensity was noted in penile vessels but not in the testes and kidney. We concluded that measurement of flow intensity in the penis as well as the changes in heart rate as the male’s reaction to the BSP can be useful in research concerning sex pheromones in dogs. [Copyright &y& Elsevier]

Domestic cats are frequently used as a research model for felid species that are threatened with extinction. Until now, the development of feline embryos has been evaluated using both classical observation methods and time-lapse monitoring (TLM). Blastocyst collapse observed using time-lapse cinematography is used as a predictor of blastocyst quality and is closely related to implantation potential. The aim of this study was to determine the relationship between the quality of domestic cat blastocysts obtained after in vitro fertilization and the frequency and duration of collapse, and of hatching. There was a significant difference in the average number of collapses and weak contractions between good and poor quality blastocysts. There was no significant difference between hatching and non-hatching blastocysts in terms of blastocyst cavity formation time or average number and duration of collapse. These results showed that the time of cavity formation was not related to blastocyst quality. The number of collapses and the occurrence of hatching were positively related to blastocyst quality, and poor quality blastocysts have, as a consequence, a reduced potential for implantation. TLM plays a significant role in cat embryo evaluation. • Collapses of cat blastocyst cavity. • Cat blastocyst quality assessment. • Cat embryo development. [ABSTRACT FROM AUTHOR]

The aim of this study was to analyze the influence of the type of spermatozoa and of different sperm abnormalities on fertilization and embryo development after ICSI in cats. In Exp I, ICSI was performed using urethral or epididymal spermatozoa collected from 7 tomcats. In Exp. II, epididymal spermatozoa from 16 cats were used for ICSI and an epididymal spermatozoon exhibiting no abnormalities or one with an abnormality was microinjected into an oocyte. Exp. I was performed in 14 replicates and Exp. II was performed in 20 replicates. In both experiments the number of cleaved oocytes, the number of embryos at the morula stage and the number of embryos at the blastocyst stage were evaluated at 24 h, and at 6 and 7 days after ICSI, respectively, and compared between experimental groups. No statistically significant differences (P > 0.05) were observed, either for Exp. I or for Exp. II. The average cleavage rate was 60.2%, morula rate 62.3% and blastocyst rate 19.2% in Exp. I and 51.6%, 66.8% and 25.8% in Exp. II, respectively. The study confirmed that both urethral and epididymal spermatozoa can be used for in vitro fertilization in cats and proved the usefulness of the ICSI method in the case of teratozoospermic males. The study showed that even in severe cases, when almost no normal spermatozoa can be found in the semen, it is possible to obtain embryos using abnormal sperm cells with the same chance of success as for normal spermatozoa. • Feline urethral and epididymal semen are equally suitable for in vitro fertilization by ICSI method. • Sperm abnormalities do not affect fertilization and embryo development after ICSI in the domestic cat. • It is possible to obtain feline embryos by ICSI in the case of very severe teratospermia using abnormal spermatozoon. [ABSTRACT FROM AUTHOR]

Abstract Steroidogenic activity in the equine ovary from birth to puberty has been poorly investigated. This study aimed to examine the capability of the ovarian follicles of prepubertal and pubertal fillies to produce steroid hormones and to evaluate the expression and cellular localization of androgen receptor (AR) in their ovaries. The ovaries of 6–18 month-old fillies were divided into two groups: prepubertal (PrP) – without preovulatory follicle (pF) and corpus luteum (CL), and ovulating/postpubertal (Ov/pB) – with pF and/or CL in at least one of the gonads. Adult mares (Me) were used as a control. The concentration of progesterone (P4), testosterone (T) and estradiol (E2) in follicular fluid (FF) was measured by radioimmunoassay. AR distribution was assessed by immunohistochemistry, while AR protein expression was examined by Western blot analysis. In the female groups, E2 concentration in FF of small follicles (<10 mm) was low and increased with the diameter of the follicle reaching the greatest value in pF (Ov/pB and Me group). In follicles (11–30 mm) of PrP fillies, the concentration of E2 was similar to that from Ov/pB fillies, but less than half (P < 0.05) than in Me follicles. In FF from all classes of follicles of Ov/pB fillies, the concentration of all steroids was similar to that in Me. AR immunolocalization, predominantly nuclear, was observed in all types of follicular cells (granulosa and theca cells) as well as in stroma and luteal cells. The pattern of staining was dependent on the follicle size and the group of females. In smaller antral follicles and in pF, the nuclear AR staining in granulosa cells was stronger than that found in follicles of 21–25 mm. In theca interna cells of pF, both nuclear and faint cytoplasmic reactions were seen. In luteal cells, AR labeling was noted in the nuclei and the cytoplasm: the strongest one in the early CL and almost negative in the late CL. AR protein expression in filly and mare ovarian tissues was confirmed by Western blot analysis and detected as a single band at approximately 110 kDa. In summary, the ovaries of fillies aged at least 6 months are capable of active steroidogenesis. ARs are present either in the cell nuclei or cytoplasm of all compartments of the equine ovary. AR expression in follicular and stroma cells may indicate the sensitivity of the filly ovarian tissue to androgens, the impact of androgens on folliculogenesis and the development of the equine ovary via a receptor-mediated pathway. Highlights • The presence of P4, T and E2 in FF of fillies ovarian follicles confirms their capability of active steroidogenesis. • AR protein was expressed in follicular, luteal and stromal cells of the equine ovary. • AR was immunolocalized both in the cell nuclei and cytoplasm of all ovarian compartments. • Expression of AR in equine ovarian cells indicates their sensitivity to androgens, which may influence folliculogenesis. [ABSTRACT FROM AUTHOR]

Abstract: Background: There are few experimental models of heart failure (HF) in large animals, despite structural and functional similarities to human myocardium. We have developed a porcine model of chronic tachycardia-induced cardiomyopathy. Methods: Homogenous siblings of White Large breed swine (n=6) underwent continuous right ventricular (RV) pacing at 170bpm; 2 subjects served as controls. In the course of RV pacing, animals developed a clinical picture of HF and were presented for euthanasia at subsequent stages: mild, moderate and end-stage HF. Left ventricle (LV) sections were analyzed histologically and relative ANP, BNP, phospholamban and sarcoplasmic reticulum calcium ATPase 2a transcript levels in LV were quantified by real time RT-PCR. Results: In the course of RV pacing, animals demonstrated reduced exercise capacity (time of running until being dyspnoeic: 6.6±0.5 vs. 2.4±1.4min), LV dilatation (LVEDD: 4.9±0.4 vs. 6.7±0.4cm), impaired LV systolic function (LVEF: 69±8 vs. 32±7 %), (all baseline vs. before euthanasia, all p<0.001). LV tissues from animals with moderate and end-stage HF demonstrated local foci of interstitial fibrosis, congestion, cardiomyocyte hypertrophy and atrophy, which was not detected in controls and mild HF animals. The up-regulation of ANP and BNP and a reduction in a ratio of sarcoplasmic reticulum calcium ATPase 2a and phospholamban in failing myocardium were observed as compared to controls. Conclusions: In pigs, chronic RV pacing at relatively low rate can be used as an experimental model of HF, as it results in a gradual deterioration of exercise tolerance accompanied by myocardial remodeling confirmed at subcellular level. [Copyright &y& Elsevier]

Microorganisms with a high natural ability to degrade xenobiotics, which are usually characterized by a diverse metabolism and unique features, can be used as natural biocatalysts in wastewater treatment or bioaugmentation processes. The immobilization of such strains greatly increases their stability and degradation properties. The most critical issues in designing whole-cell immobilization systems are selecting the suitable carrier and determining the bacterial features that will promote effective immobilization. In this study, we immobilized the paracetamol-degrading Pseudomonas moorei KB4 strain on the bacterial cellulose disks that were produced by Komagataeibacter xylinus E-89370. The KB4 strain immobilized on the cellulose degraded 150 mg L−1 of paracetamol in three series of 50 mg L−1 in each cycle. The average protein concentration and dehydrogenase activity increased after the degradation of each dose. The specific activity (U mg−1 of protein) of the main enzymes involved in the degradation pathway was 0.22 for deaminase, 5.1 for acylamidohydrolase and 3.49 for ring-cleavage hydroquinone 1,2-dioxygenase. The relative expression level of the genes encoding deaminases and acylamidohydrolases increased in the presence of paracetamol, though more prominently in the immobilized than in the free cells. Image 1 • Pseudomonas moorei KB4 exhibits the most critical traits for immobilization. • Immobilized KB4 strain degrades 150 mg L−1 of paracetamol. • Immobilized cells transform paracetamol via p -aminophenol and hydroquinone. • Paracetamol exposure up-regulated expression of genes encoded degradative enzymes. [ABSTRACT FROM AUTHOR]