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Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system Texto completo
2021
Blanco, Federico Carlos | Vazquez, Cristina Lourdes | García, Julia Sabio y | Rocha, Rosana Valeria | Gravisaco, María José | Forrellad, Marina Andrea | Magistrelli, Giovanni | Bigi, Fabiana
Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system Texto completo
2021
Blanco, Federico Carlos | Vazquez, Cristina Lourdes | García, Julia Sabio y | Rocha, Rosana Valeria | Gravisaco, María José | Forrellad, Marina Andrea | Magistrelli, Giovanni | Bigi, Fabiana
Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) are cytokines widely used in ex vivo monocyte differentiation experiments, vaccine formulations and disease treatment. The aim of this study was to produce recombinant bovine GM-CSF and IL-4 in an episomal expression system that conserves the postransductional modification of the native proteins and to use the products to differentiate bovine monocytes into dendritic cells. The recombinant proteins rGM-CSF and rIL-4 were expressed in PEAKrapid CRL-2828 human kidney cells, ATCC CRL-2828. The functional activity of the recombinant cytokines was monitored by registering morphological changes in bovine monocytes and assessing the expression of CD14 upon incubation with them. Both recombinant proteins were detected in the cell culture supernatant of transfected cells. Culture supernatants of transfected cells induced in bovine monocytes morphological changes that resemble macrophages or dendritic cells. In addition, bovine cells treated with rGM-CSF and rIL-4 showed reduced expression of the macrophage surface marker CD14 compared with untreated cells. This effect indicates the expected differentiation. The expression of the cytokines was stable after many successive cell passages and a freeze/thaw cycle. The semi-stable mammalian episomal expression system used in this study allowed us to easily produce functional bovine rGM-CSF and rIL-4 without the need for protein purification steps.
Mostrar más [+] Menos [-]Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system Texto completo
2021
Blanco Federico Carlos | Vazquez Cristina Lourdes | García Julia Sabio y | Rocha Rosana Valeria | Gravisaco María José | Forrellad Marina Andrea | Magistrelli Giovanni | Bigi Fabiana
Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) are cytokines widely used in ex vivo monocyte differentiation experiments, vaccine formulations and disease treatment. The aim of this study was to produce recombinant bovine GM-CSF and IL-4 in an episomal expression system that conserves the postransductional modification of the native proteins and to use the products to differentiate bovine monocytes into dendritic cells.
Mostrar más [+] Menos [-]A pilot study to establish an ovalbumin-induced atopic dermatitis minipig model Texto completo
2021
Kim, Yŏng-gyu | Lee, Ju Young | Hwang, Jeong Ho | Suh, Han Na
A pilot study to establish an ovalbumin-induced atopic dermatitis minipig model Texto completo
2021
Kim, Yŏng-gyu | Lee, Ju Young | Hwang, Jeong Ho | Suh, Han Na
Because minipig skin is similar to human skin in anatomy and physiology, establishing an atopic dermatitis (AD) minipig model seems meaningful. We applied 1-fluoro-2,4-dinitrobenzene (DNFB) or ovalbumin onto the back skin of five Yucatan minipigs aged 8–10 months and 19 kg in median weight. Two minipigs with the same parameters served as controls. Both DNFB and ovalbumin mediated epithelial hyperplasia, spongiosis, and immune cell infiltration in the dermis, which is a typical histopathological feature of AD. Moreover, AD upregulated the Th1- and Th2-related cytokine expressions in DNFB- or in ovalbumin-treated skin. Notably, AD-induced minipigs exhibited greater cytokine serum concentrations. Histopathological finding and cytokine analysis revealed that DNFB or ovalbumin mediates AD. However, ovalbumin-treated minipig is a more reliable and precise AD model owing to the DNFB-induced severe skin damage. In summary, ovalbumin-treated skin shows similar AD as human in histopathological and molecular analysis.
Mostrar más [+] Menos [-]A pilot study to establish an ovalbumin-induced atopic dermatitis minipig model Texto completo
2021
Kim Young Kyu | Lee Ju Young | Hwang Jeong Ho | Suh Han Na
Because minipig skin is similar to human skin in anatomy and physiology, establishing an atopic dermatitis (AD) minipig model seems meaningful.
Mostrar más [+] Menos [-]Clinical use of the parasympathetic tone activity index as a measurement of postoperative analgaesia in dogs undergoing ovariohysterectomy Texto completo
2021
Clinical use of the parasympathetic tone activity index as a measurement of postoperative analgaesia in dogs undergoing ovariohysterectomy Texto completo
2021
While the current tools to assess canine postoperative pain using physiological and behavioural parameters are reliable, an objective method such as the parasympathetic tone activity (PTA) index could improve postoperative care. The aim of the study was to determine the utility of the PTA index in assessing postoperative analgaesia. Thirty healthy bitches of different breeds were randomly allocated into three groups for analgaesic treatment: the paracetamol group (GPARAC, n = 10) received 15 mg/kg b.w., the carprofen group (GCARP, n = 10) 4 mg/kg b.w., and the meloxicam group (GMELOX, n = 10) 0.2 mg/kg b.w. for 48 h after surgery. GPARAC was medicated orally every 8 h, while GCARP and GMELOX were medicated intravenously every 24 h. The PTA index was used to measure the analgaesia–nociception balance 1 h before surgery (baseline), and at 1, 2, 4, 6, 8, 12, 16, 20, 24, 36, and 48 h after, at which times evaluation on the University of Melbourne Pain Scale (UMPS) was made. The baseline PTA index was 65 ± 8 for GPARAC, 65 ± 7 for GCARP, and 62 ± 5 for GMELOX. Postoperatively, it was 65 ± 9 for GPARAC, 63 ± 8 for GCARP, and 65 ± 8 for GMELOX. No statistically significant difference existed between baseline values or between values directly after treatments (P = 0.99 and P = 0.97, respectively). The PTA index showed a sensitivity of 40%, specificity of 98.46% and a negative predictive value of 99.07%. Our findings suggest that the PTA index measures comfort and postoperative analgaesia objectively, since it showed a clinical relationship with the UMPS.
Mostrar más [+] Menos [-]Clinical use of the parasympathetic tone activity index as a measurement of postoperative analgaesia in dogs undergoing ovariohysterectomy Texto completo
2021
Hernández-Avalos Ismael | Valverde Alex | Antonio Ibancovichi-Camarillo José | Sánchez-Aparicio Pedro | Recillas-Morales Sergio | Rodríguez-Velázquez Desiderio | Osorio-Avalos Jorge | Armando Magdaleno-Torres Luis | Chavez-Monteagudo Julio | Manuel Acevedo-Arcique Carlos
While the current tools to assess canine postoperative pain using physiological and behavioural parameters are reliable, an objective method such as the parasympathetic tone activity (PTA) index could improve postoperative care. The aim of the study was to determine the utility of the PTA index in assessing postoperative analgaesia.
Mostrar más [+] Menos [-]In vivo study of the oestrogenic activity of milk Texto completo
2021
Radko, Lidia | Posyniak, Andrzej
In vivo study of the oestrogenic activity of milk Texto completo
2021
Radko, Lidia | Posyniak, Andrzej
Milk has been suggested to be a possible source of oestrogenically active compounds. In order to assess the health risk for milk consumers and ensure the safety of this staple part of the human diet, it is important to study the effect of xenooestrogen mixtures present in milk. This investigation used the available in vivo model to learn to what extent such compounds may be endocrine disruptors. The recommended immature golden hamster uterotrophic bioassay was chosen. A total of 132 animals were divided into nine groups of experimental animals and positive and negative control groups, each of 12 animals. The experimental females received ad libitum either one of five samples of raw cow’s milk from individual animals or one of four samples of pasteurised or ultra-high temperature treated cow’s milk as retail products. After 7 days, the animals were sacrificed and necropsied. Uterine weight increases were measured as the endpoint of oestrogenic activity in milk. The milk samples from individual cows and the retail milk samples did not show oestrogenic activity. However, in three groups, decreased uterine weights were observed. Considering that milk supplies are beneficial to health, contamination in this food should be avoided. There is a need for further animal experiments and epidemiological studies are warranted to evaluate any causative role of milk in human endocrinological disorders.
Mostrar más [+] Menos [-]In vivo study of the oestrogenic activity of milk Texto completo
2021
Radko Lidia | Posyniak Andrzej
Milk has been suggested to be a possible source of oestrogenically active compounds. In order to assess the health risk for milk consumers and ensure the safety of this staple part of the human diet, it is important to study the effect of xenooestrogen mixtures present in milk. This investigation used the available in vivo model to learn to what extent such compounds may be endocrine disruptors.
Mostrar más [+] Menos [-]Acrylamide-induced changes of granulopoiesis in porcine bone marrow Texto completo
2021
Grzybowska, Dominika | Snarska, Anna
Acrylamide-induced changes of granulopoiesis in porcine bone marrow Texto completo
2021
Grzybowska, Dominika | Snarska, Anna
Due to the widely documented and diverse toxic effects of acrylamide, the authors decided to evaluate the impact of high and low doses of this compound on the process of granulopoiesis in porcine bone marrow. The experiment was conducted on 15 Danish Landrace pigs at the age of 8 weeks. The animals were randomly assigned into three equal groups (n = 5). Control animals received empty gelatine capsules as placebo. Animals in the first experimental group (the LD group) received a low dose of acrylamide of 0.5 μg/kg b.w./day, and animals in the second experimental group (the HD group) received a tenfold higher dose of acrylamide of 5 μg/kg b.w./day. Placebo and acrylamide capsules were administered with feed every morning for 28 days. Bone marrow was collected into tubes without an anticoagulant twice – before the first capsule administration (day 0) and on the 28ᵗʰ day of the study. After drying and staining, bone marrow smears were subjected to detailed cytological evaluation under a light microscope. Changes in cell morphology, i.e. degenerative changes in the cellular nuclei, were observed in both experimental groups. Both low and high doses of acrylamide decreased the number of segmented eosinophils, neutrophilic and segmented metamyelocytes, neutrophils, as well as basophils and basophilic metamyelocytes. Acrylamide at doses of 0.5 μg/kg b.w./day and 5 μg/kg b.w./day clearly influences porcine granulopoiesis.
Mostrar más [+] Menos [-]Acrylamide-induced changes of granulopoiesis in porcine bone marrow Texto completo
2021
Grzybowska Dominika | Snarska Anna
Due to the widely documented and diverse toxic effects of acrylamide, the authors decided to evaluate the impact of high and low doses of this compound on the process of granulopoiesis in porcine bone marrow.
Mostrar más [+] Menos [-]Comparative expression analysis of inflammatory and immune-related genes in cattle during acute infection with foot-and-mouth disease virus in Egypt Texto completo
2021
El Nahas, Abeer F. | Abd El Naby, Walaa S.H. | Khatab, Shymaa A. | Fergany, Al-Zahraa A. | Rashed, Rashed R.
Comparative expression analysis of inflammatory and immune-related genes in cattle during acute infection with foot-and-mouth disease virus in Egypt Texto completo
2021
El Nahas, Abeer F. | Abd El Naby, Walaa S.H. | Khatab, Shymaa A. | Fergany, Al-Zahraa A. | Rashed, Rashed R.
Foot-and-mouth disease is a highly infectious viral disease affecting all cloven-footed domestic animals. The three foot-and-mouth disease virus (FMDV) serotypes A, O and SAT2 are at present the greatest threat to susceptible animals in Egypt. The aim of the present study was, for the host factors associated with different FMDV infections in cattle during the acute phase, to compare these factors’ influence on the expression of the IL-10, TLR-2, TNF-α, CXCL10, CD48, NFATC4 and IFNG inflammatory and immune-related genes. Vesicular fluid and epithelium samples were obtained from at least three infected cattle on the same affected farm during three different FMDV outbreaks and were used for serotyping of the virus and for expression analysis of host genes. A two-step RT-PCR was used for diagnosis of the virus with primers specific for each serotype. In quantitative PCR analysis, the expression patterns of TLR-2 and IFNG were prominent, while NFATC4 expression was absent in all FMDV-infected cattle. The highest expression of CD48 was associated with increased expression of other inflammatory and immune-related genes (IL-10, TLR-2, TNF-α and IFNG), which may be an indication of rapid virus clearance. The use of vesicular fluid and epithelium for investigation of viral and immune-related gene expression levels in acute FMDV infection is possible. Host-dependent variation in the expression of the studied genes was observed in different FMDV serotype outbreaks.
Mostrar más [+] Menos [-]Comparative expression analysis of inflammatory and immune-related genes in cattle during acute infection with foot-and-mouth disease virus in Egypt Texto completo
2021
El Nahas Abeer F. | Abd El Naby Walaa S.H. | Khatab Shymaa A. | Fergany Al-Zahraa A. | Rashed Rashed R.
Foot-and-mouth disease is a highly infectious viral disease affecting all cloven-footed domestic animals. The three foot-and-mouth disease virus (FMDV) serotypes A, O and SAT2 are at present the greatest threat to susceptible animals in Egypt. The aim of the present study was, for the host factors associated with different FMDV infections in cattle during the acute phase, to compare these factors’ influence on the expression of the IL-10, TLR-2, TNF-α, CXCL10, CD48, NFATC4 and IFNG inflammatory and immune-related genes.
Mostrar más [+] Menos [-]Phytoncides in the prevention and therapy of blackhead disease and their effect on the turkey immune system Texto completo
2021
Tykałowski, Bartłomiej | Śmiałek, Marcin | Kowalczyk, Joanna | Dziewulska, Daria | Stenzel, Tomasz | Koncicki, Andrzej
Phytoncides in the prevention and therapy of blackhead disease and their effect on the turkey immune system Texto completo
2021
Tykałowski, Bartłomiej | Śmiałek, Marcin | Kowalczyk, Joanna | Dziewulska, Daria | Stenzel, Tomasz | Koncicki, Andrzej
Turkey histomonosis poses a serious threat to poultry production due to the ban on the use of effective drugs. The aim of the study was to evaluate the influence of a phytoncidal feed supplement on the course of histomonosis. The preparation was also analysed for immunomodulatory properties. Clinical observations and production monitoring were conducted in a flock of turkeys with histomonosis from their 11ᵗʰ to 56ᵗʰ weeks of life which were treated with the adiCoxSᴼᴸPF soluble supplement in a dose of 2.5 mL/L water. Later the preparation was used in a preventive dose (1 mL/L). The influence on the immune system was evaluated in broiler turkeys having been given adiCoxSᴼᴸPF for 3 days in doses of 1 or 3 mL/L. The T and B lymphocyte percentages in turkey blood and spleen tissue were analysed with flow cytometry. ELISA was implemented to evaluate antibody titres after Ornithobacterium rhinotracheale vaccination, and biochemical analyses were performed to evaluate the supplement’s safety. AdiCoxSᴼᴸPF was found effective in therapy and prevention of histomonosis. Additionally, adiCoxSᴼᴸPF stimulated both humoral and cell-mediated immune mechanisms, without impairing the functions of internal organs. The treated turkeys also yielded better production results (eggs/hen, fertility, and hatchability). AdiCoxSᴼᴸPF possesses immunomodulatory properties and it can be used successfully in the prevention and therapy of histomonosis in turkeys.
Mostrar más [+] Menos [-]Phytoncides in the prevention and therapy of blackhead disease and their effect on the turkey immune system Texto completo
2021
Tykałowski Bartłomiej | Śmiałek Marcin | Kowalczyk Joanna | Dziewulska Daria | Stenzel Tomasz | Koncicki Andrzej
Turkey histomonosis poses a serious threat to poultry production due to the ban on the use of effective drugs. The aim of the study was to evaluate the influence of a phytoncidal feed supplement on the course of histomonosis. The preparation was also analysed for immunomodulatory properties.
Mostrar más [+] Menos [-]Assessment of the morphometry of heads of normal sperm and sperm with the Dag defect in the semen of Duroc boars Texto completo
2021
Banaszewska, Dorota | Andraszek, Katarzyna
Assessment of the morphometry of heads of normal sperm and sperm with the Dag defect in the semen of Duroc boars Texto completo
2021
Banaszewska, Dorota | Andraszek, Katarzyna
The Dag defect is one of the primary morphological defects in sperm correlating with reduced fertility. This defect is found in the spermatozoa of many livestock species. The aim of the study was to assess the morphometry of the heads of normal sperm and sperm with the Dag defect in the semen of Duroc breeding boars. Sperm morphology was examined in ten ejaculates each from 12 Duroc boars. In total, 3,600 morphologically normal sperm and 838 sperm with the Dag defect were evaluated. The area, perimeter, length and width of the sperm head were measured and these basic morphometric parameters were used to calculate four additional shape indices characterising the sperm head, i.e. ellipticity, elongation, roughness and regularity. Sperm with this defect had markedly smaller heads, 0.32 μm shorter and 0.19 μm narrower than the heads of sperm with normal morphological structure. The heads of sperm with the Dag defect also had a 1.1μm smaller perimeter and a 2.5 μm² smaller surface area than the heads of morphologically normal sperm. The Dag defect is found in boar sperm irrespective of the age of the individual. It affects the morphology of the sperm head.
Mostrar más [+] Menos [-]Assessment of the morphometry of heads of normal sperm and sperm with the Dag defect in the semen of Duroc boars Texto completo
2021
Banaszewska Dorota | Andraszek Katarzyna
The Dag defect is one of the primary morphological defects in sperm correlating with reduced fertility. This defect is found in the spermatozoa of many livestock species. The aim of the study was to assess the morphometry of the heads of normal sperm and sperm with the Dag defect in the semen of Duroc breeding boars.
Mostrar más [+] Menos [-]Protective effects of bisoprolol against cadmium-induced myocardial toxicity through inhibition of oxidative stress and NF-κΒ signalling in rats Texto completo
2021
Liu, Jinhua | Xie, Ying | Han, Zhujun | Wang, Hailong | Xu, Wenhu
Protective effects of bisoprolol against cadmium-induced myocardial toxicity through inhibition of oxidative stress and NF-κΒ signalling in rats Texto completo
2021
Liu, Jinhua | Xie, Ying | Han, Zhujun | Wang, Hailong | Xu, Wenhu
The aim of the study was to investigate the mitigative effects of bisoprolol (BIS) in cadmium-induced myocardial toxicity on oxidative stress and its inhibitive effect on nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) signalling in rats. Male albino Wistar rats were assigned to control, Cd, BIS 2 (2 mg/kg b.w.) and BIS 8 (8 mg/kg b.w.) groups with nine rats in each. Over four weeks, the control group was administered 1% gum acacia, all other groups received 3mg/kg b.w. CdCl₂ dissolved in distilled water, and the BIS groups were additionally given bisoprolol in gum acacia. Blood samples were collected for biochemical estimations. Blood pressure and serum biomarker (lactate dehydrogenase, aspirate transaminase, alanine transferase and creatine kinase-MB, enzyme (superoxide dismutase, lipid hydroxy peroxidase, catalase and malondialdehyde), and tumour necrosis factor alpha (TNF-α) concentrations were measured. Western blot analysis was conducted for NF-κB and glutathione S-transferase (GST). After sacrificing the rats, cardiac tissue samples were examined histopathologically. Our findings pointed to a significant decrease (P < 0.05) in the studied serum biomarkers and levels of the relevant enzymes in the BIS 8 group compared to the Cd group. A significant decrease (P < 0.05) in NF-kB p65 expression and TNF-α levels was noted in the BIS 8 group relative to the BIS 2 and Cd groups, indicating a reduction at a higher dose. In microscopy, histopathological changes in the cardiac muscles of the BIS 8 group were evident compared to those of the Cd group. BIS seemed to have protective effects against cardiac injury induced by cadmium and could be considered a novel therapeutic drug and prognostic biomarker in the pathology of the many cardiovascular diseases caused by heavy metal intake.
Mostrar más [+] Menos [-]Protective effects of bisoprolol against cadmium-induced myocardial toxicity through inhibition of oxidative stress and NF-κΒ signalling in rats Texto completo
2021
Liu Jinhua | Xie Ying | Han Zhujun | Wang Hailong | Xu Wenhu
The aim of the study was to investigate the mitigative effects of bisoprolol (BIS) in cadmium-induced myocardial toxicity on oxidative stress and its inhibitive effect on nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) signalling in rats.
Mostrar más [+] Menos [-]Antimicrobial resistance and virulence factor gene profiles of Enterococcus spp. isolated from giant panda oral cavities Texto completo
2021
Zhong, Rui | Zhou, Ziyao | Liu, Haifeng | Zhong, Zhijun | Peng, Guangneng
Antimicrobial resistance and virulence factor gene profiles of Enterococcus spp. isolated from giant panda oral cavities Texto completo
2021
Zhong, Rui | Zhou, Ziyao | Liu, Haifeng | Zhong, Zhijun | Peng, Guangneng
The objective of this study was to determine the prevalence and characteristics of antimicrobial-resistant Enterococcus faecalis and E. faecium isolated from the oral cavities of captive giant pandas in China. The virulence-associated determinant and antimicrobial resistance genes were detected and antimicrobial susceptibility tests were performed on 54 strains of each bacterium. All isolates showed 100% multidrug resistance. E. faecalis isolates showed a higher percentage of strains resistant to gentamicin (48.1%), vancomycin (55.6%), linezolid (100%), and streptomycin (33.3%) than E. faecium isolates. The resistance genes of Enterococcus spp. were present to highly varying extents according to antibiotic type, their presence breaking down for E. faecalis and E. faecium respectively as aac(6')/aph(2″) 5.56% and 5.56%; aph(3')-Ⅲ 0% and 14.81%; ant(6)-I 0% and 3.7%; ant(4')-Ia 0% and 64.81%; tetL 20.37% and 100%; vanA 92.59% and 46.3%; vanB 0% and 0%; cfr 0% and 90.74%; optrA 96.3% and 3.7%; blaZ 0% and 1.85%; blaTEM 0% and 0%; tetA 20.37% and 0%; tetC 24.07% and 100%; tetM 0% and 0%; ermA 12.96% and 100%; ermB 5.56% and 3.7%; and ermC 0% and 1.85%.Virulence-associated determinants were detected in this research, which typically include efaA, gelE, asa1, ace, cylA, esp and hyl; however, the latter three were not detected. High proportions of the isolates carried the efaA, gelE, asa1, and ace genes. Respectively for E. faecalis and E. faecium their detection was efaA 98.1% and 85.2%; gelE 98.1% and 87%; asa1 92.6% and 87%; and ace 87% and 85.2%. This is the first study on the potential disease risk and antimicrobial-resistant characteristics of E. faecalis and E. faecium isolates in giant panda oral cavities. The results of this study show that the antimicrobial resistance rate of Enterococcus spp. isolated from the oral cavity of captive pandas is very high, and thus needs to be monitored.
Mostrar más [+] Menos [-]Antimicrobial resistance and virulence factor gene profiles of Enterococcus spp. isolated from giant panda oral cavities Texto completo
2021
Zhong Rui | Zhou Ziyao | Liu Haifeng | Zhong Zhijun | Peng Guangneng
The objective of this study was to determine the prevalence and characteristics of antimicrobial-resistant Enterococcus faecalis and E. faecium isolated from the oral cavities of captive giant pandas in China.
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