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Purification of myeloperoxidase from equine polymorphonuclear leucocytes. 全文
1998
Mathy, Marianne | Bourgeois, E. | Grulke, Sigrid | Deby, Ginette | Caudron, I. | Deby, C. | Lamy, Maurice | Serteyn, Didier
peer reviewed | Increases of plasma concentrations of neutrophil myeloperoxidase (MPO) can be used as markers of polymorphonuclear leucocytes (PMN) activation in pathological situations (sepsis, acute lung injury, acute inflammation). To develop an assay for measurement of plasma MPO in horses during the above-mentioned infectious and inflammatory conditions, MPO was purified from equine PMN isolated from blood anticoagulated with citrate. PMN were extracted in a saline milieu (0.2 M Na acetate, 1 M NaCl, pH 4.7) to eliminate most of cellular proteins. Pellets were then extracted in the same buffer containing cationic detergent (1% cetyltrimethyl ammonium bromide). The supernatant was further purified by ion exchange chromatography (Hiload S Sepharose HP column 0.5 x 26 cm, equilibrated with 25 mM Na acetate, 0.2 M NaCl, pH 4.7) with a NaCl gradient (until 1 M). Most of the peroxidase activity of MPO (spectrophotometrically measured by the oxidation of orthodianisidine by hydrogen peroxide) was eluted at 0.65 M NaCl. MPO was further purified by gel filtration chromatography (Sephacryl S 200 column 2.6 x 42 cm with 25 mM Na acetate, 0.2 M NaCl, pH 4.7). MPO (specific activity: 74.3 U/mg) was obtained with a yield of 30% from the detergent extraction supernatant. Electrophoresis (non-reducing conditions) showed 3 bands identified, by comparison with human MPO, (i) the mature tetrameric enzyme (150 kDa) with 2 light and 2 heavy subunits, (ii) the precursor form (88 kDa) and (iii) a form of the heavy subunit without the prosthetic heme group (40 kDa). The mature enzyme and its precursor were glycosylated and possessed peroxidase activity. Equine MPO showed strong similarities with human and bovine MPO, with an absorption peak at 430 nm (Soret peak) characteristic of ferrimyeloperoxidase. Enzymatic activity was pH dependent (optimal value at pH 5.5).
显示更多 [+] 显示较少 [-]Establishment of a new canine inflammatory mammary carcinoma cell line and analysis of its cystine-glutamate transporter subunit expression 全文
2022
Itoh, Harumichi | Naruse, Ryo | Tani, Kenji | Sunahara, Hiroshi | Nemoto, Yuki | Nakaichi, Munekazu | Iseri, Toshie | Horikirizono, Hiro | Itamoto, Kazuhito
Establishment of a new canine inflammatory mammary carcinoma cell line and analysis of its cystine-glutamate transporter subunit expression 全文
2022
Itoh, Harumichi | Naruse, Ryo | Tani, Kenji | Sunahara, Hiroshi | Nemoto, Yuki | Nakaichi, Munekazu | Iseri, Toshie | Horikirizono, Hiro | Itamoto, Kazuhito
Inflammatory mammary carcinoma (IMC) is a rare disease with a poor prognosis and one affecting dogs. Inflammatory breast carcinoma (IBC) is a subtype of malignant breast cancer in humans with a high degree of malignancy and a similarly poor prognosis. Since the clinical symptoms and prognoses of both are similar, canine IMC has been considered as a model of human IBC. In this study, we newly established a stable IMC-derived cell line from a patient at the Yamaguchi University Animal Medical Center in Japan. The patient was a female toy poodle presenting with an inflamed mammary gland, which was diagnosed as IMC. The cell line was established from a tissue biopsy. Surface antigen marker (CD24 and CD44) expression was determined. Cystine/glutamate antiporter (xCT) expression was determined by Western blotting, flow cytometry and fluorescence immunostaining, and sulfasalazine was administered to ascertain if it suppressed xCT expression. Stem cell marker (Nanog, Sox2, Myc and Klf4) expression and aldehyde dehydrogenase (ALDH) activity were also investigated. The cultured cells showed xCT, and its suppression showed downregulation of stem cell markers and ALDH activity. Stable cell proliferation was verified. A new canine IMC-derived cell line was established. In the future, we aim to study the effect of xCT on the maintenance of cancer stem cell properties in canine tumours, and propose a new therapeutic method for the treatment of canine IMC by targeting xCT.
显示更多 [+] 显示较少 [-]Establishment of a new canine inflammatory mammary carcinoma cell line and analysis of its cystine-glutamate transporter subunit expression 全文
2022
Itoh Harumichi | Naruse Ryo | Tani Kenji | Sunahara Hiroshi | Nemoto Yuki | Nakaichi Munekazu | Iseri Toshie | Horikirizono Hiro | Itamoto Kazuhito
Inflammatory mammary carcinoma (IMC) is a rare disease with a poor prognosis and one affecting dogs. Inflammatory breast carcinoma (IBC) is a subtype of malignant breast cancer in humans with a high degree of malignancy and a similarly poor prognosis. Since the clinical symptoms and prognoses of both are similar, canine IMC has been considered as a model of human IBC. In this study, we newly established a stable IMC-derived cell line from a patient at the Yamaguchi University Animal Medical Center in Japan.
显示更多 [+] 显示较少 [-]Characterisation of a new molecule based on two E2 sequences from bovine viral diarrhoea-mucosal disease virus fused to the human immunoglobulin Fc fragment 全文
2021
González Pose, Alaín | Montesino Seguí, Raquel | Maura Pérez, Rafael | Hugues Salazar, Florence | Cabezas Ávila, Ignacio | Altamirano Gómez, Claudia | Sánchez Ramos, Oliberto | Roberto Toledo, Jorge
Characterisation of a new molecule based on two E2 sequences from bovine viral diarrhoea-mucosal disease virus fused to the human immunoglobulin Fc fragment 全文
2021
González Pose, Alaín | Montesino Seguí, Raquel | Maura Pérez, Rafael | Hugues Salazar, Florence | Cabezas Ávila, Ignacio | Altamirano Gómez, Claudia | Sánchez Ramos, Oliberto | Roberto Toledo, Jorge
Proper conformational arrangement of the E2 molecules of bovine viral diarrhoea-mucosal disease virus (BVD-MDV) is crucial to obtain an effective recombinant vaccine candidate against the disease. In this study, we characterised a new molecule composed of two distinct sequences of the E2 glycoprotein of BVD-MDV and the Fc fragment of human immunoglobulin (BVDE2Fc). The chimaeric protein was expressed in mammalian cell lines of different species by adenoviral transduction and purified by immobilised metal-affinity chromatography. The N-glycans were profiled by HPLC, and the BVDE2Fc immunogenicity was assessed in male mice. The antigen-antibody reactions were evaluated by ELISA. The MDBK cell line was selected from among five for the final production of BVDE2Fc. After purification to over 90%, the N-glycan profile showed neutral and complex oligosaccharides. The mouse immunisation induced a strong humoral response, which produced antibodies able to attach to conformational epitopes on E2 molecules, while the Fc fragment barely contributed to the immune response. Additionally, BVDE2Fc attached to antibodies from bovine sera positive to distinct BVD-MDV subtypes, whereas the loss of BVDE2Fc structure during the deglycosylation process considerably diminished those interactions. These results demonstrate that the structure of E2 molecules arranged in tandem and attached to an Fc fragment could represent a viable design for future vaccine candidates against BVD-MD.
显示更多 [+] 显示较少 [-]Characterisation of a new molecule based on two E2 sequences from bovine viral diarrhoea-mucosal disease virus fused to the human immunoglobulin Fc fragment 全文
2021
González Pose Alaín | Montesino Seguí Raquel | Maura Pérez Rafael | Hugues Salazar Florence | Cabezas Ávila Ignacio | Altamirano Gómez Claudia | Sánchez Ramos Oliberto | Roberto Toledo Jorge
Proper conformational arrangement of the E2 molecules of bovine viral diarrhoea-mucosal disease virus (BVD-MDV) is crucial to obtain an effective recombinant vaccine candidate against the disease. In this study, we characterised a new molecule composed of two distinct sequences of the E2 glycoprotein of BVD-MDV and the Fc fragment of human immunoglobulin (BVDE2Fc).
显示更多 [+] 显示较少 [-]Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system 全文
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 全文
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.
显示更多 [+] 显示较少 [-]Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system 全文
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.
显示更多 [+] 显示较少 [-]A pilot study to establish an ovalbumin-induced atopic dermatitis minipig model 全文
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 全文
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.
显示更多 [+] 显示较少 [-]A pilot study to establish an ovalbumin-induced atopic dermatitis minipig model 全文
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.
显示更多 [+] 显示较少 [-]In vivo study of the oestrogenic activity of milk 全文
2021
Radko, Lidia | Posyniak, Andrzej
In vivo study of the oestrogenic activity of milk 全文
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.
显示更多 [+] 显示较少 [-]In vivo study of the oestrogenic activity of milk 全文
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.
显示更多 [+] 显示较少 [-]E-test determination of antifungal susceptibility of Candida species isolated from turkeys 全文
2020
Sokół, Ireneusz | Tokarzewski, Stanisław | Bobrek, Kamila | Gaweł, Andrzej
E-test determination of antifungal susceptibility of Candida species isolated from turkeys 全文
2020
Sokół, Ireneusz | Tokarzewski, Stanisław | Bobrek, Kamila | Gaweł, Andrzej
Candida species are a natural component of the intestinal tract microflora, but in favourable conditions they can cause superficial, mucosal, or even systemic candidiasis. Poultry production might be a source of human drug-resistant yeast infections, including Candida spp. The limited data concerning the antifungal susceptibility of poultry Candida isolates prompted us to carry out research to determine the susceptibility of isolates from turkey intestinal tracts. The beak cavity, crop and cloaca were swabbed of 580 turkeys from 58 flocks in western Poland. The susceptibility tests were conducted using the E-test method with amphotericin B, fluconazole, itraconazole, and voriconazole on 52 isolates of C. albicans, C. catenulata, C. glabrata, C. palmioleophila, C. rugosa, C. krusei and C. lusitaniae. All isolates were susceptible to voriconazole. According to the MIC values obtained for amphotericin B and fluconazole, all Candida spp. isolates were classified as susceptible according to the described breakpoints except for C. krusei, which was the only isolate that was amphotericin B-, fluconazole- and itraconazole-resistant. The susceptibility to itraconazole varied: 11 of the Candida isolates were susceptible (21.1%), 29 were dose-dependently susceptible (55.8%), and 12 isolates were resistant (23.1%). There are few resistant strains of Candida in turkeys, and the drug resistance varies. When Candida passes from turkeys to humans, there is a wide range of antifungal treatment options.
显示更多 [+] 显示较少 [-]E-test determination of antifungal susceptibility of Candida species isolated from turkeys 全文
2020
Sokół Ireneusz | Tokarzewski Stanisław | Bobrek Kamila | Gaweł Andrzej
Candida species are a natural component of the intestinal tract microflora, but in favourable conditions they can cause superficial, mucosal, or even systemic candidiasis. Poultry production might be a source of human drug-resistant yeast infections, including Candida spp. The limited data concerning the antifungal susceptibility of poultry Candida isolates prompted us to carry out research to determine the susceptibility of isolates from turkey intestinal tracts.
显示更多 [+] 显示较少 [-]Carbapenem-resistant Pseudomonas aeruginosa originating from farm animals and people in Egypt 全文
2019
Elshafiee Esraa A. | Nader Sara M. | Dorgham Sohad M. | Hamza Dalia A.
Carbapenem-resistant Pseudomonas aeruginosa originating from farm animals and people in Egypt 全文
2019
Elshafiee Esraa A. | Nader Sara M. | Dorgham Sohad M. | Hamza Dalia A.
Carbapenem-resistant Pseudomonas aeruginosa (CRPA) has become the leading cause of health care-associated infections. Treatment is difficult due to the lack of an effective antimicrobial therapy, and mortality is high. This study investigated the occurrence of CRPA in farm animals (buffaloes and cattle), livestock drinking water, and humans in Egypt.
显示更多 [+] 显示较少 [-]Carbapenem-resistant Pseudomonas aeruginosa originating from farm animals and people in Egypt 全文
2019
Elshafiee, Esraa A. | Nader, Sara M. | Dorgham, Sohad M. | Hamza, Dalia A.
Carbapenem-resistant Pseudomonas aeruginosa (CRPA) has become the leading cause of health care-associated infections. Treatment is difficult due to the lack of an effective antimicrobial therapy, and mortality is high. This study investigated the occurrence of CRPA in farm animals (buffaloes and cattle), livestock drinking water, and humans in Egypt. A total of 180 samples were examined: 50 faecal each from buffaloes and cattle, 30 of livestock drinking water, and 50 stool from humans. The samples were cultured on cetrimide agar and the plates were incubated aerobically at 37°C for 24 h. The isolates were examined for the presence of the blaKPC, blaOXA₋₄₈, and blaNDM carbapenemase-encoding genes using PCR and investigated for the exotoxin A (toxA) gene. The toxA gene from carbapenem- group resistant isolates was phylogenetically analysed. P. aeruginosa was isolated from buffaloes, cattle, drinking water, and humans, with occurrences of 40%, 34%, 10%, and 20%, respectively. Carbapenem resistance genes were found in 60%, 59%, 67%, and 70% in buffalo, cattle, water and human samples, respectively. The toxA gene was detected in 80% of samples. The phylogenetic analysis showed that cattle and water sequences were in one cluster and more related to each other than to human isolates. Occurrence of CRPA among farm animals, drinking water, and humans was high, reflecting the environmental origin of P. aeruginosa and highlighting contaminated water as a potential transmitter of CRPA to livestock and next to humans.
显示更多 [+] 显示较少 [-]Use of aVR and Jastrzębski algorithms in the classification of wide complex tachycardia in dogs – a preliminary study 全文
2018
Michałek, Marcin | Frydrychowski, Piotr | Adamowicz, Jakub | Sławuta, Agnieszka | Pasławska, Urszula | Noszczyk-Nowak, Agnieszka
Use of aVR and Jastrzębski algorithms in the classification of wide complex tachycardia in dogs – a preliminary study 全文
2018
Michałek, Marcin | Frydrychowski, Piotr | Adamowicz, Jakub | Sławuta, Agnieszka | Pasławska, Urszula | Noszczyk-Nowak, Agnieszka
Ventricular rhythm disturbances are a common pathology in human and veterinary medicine. In humans, the algorithmic approach is used to differentiate wide QRS complex tachycardia. The most commonly used are the aVR and Brugada algorithms as well as the ventricular tachycardia (VT) score developed by Jastrzębski and coworkers. In veterinary medicine, no such algorithms are available and the only parameter used to describe VT abnormalities is the duration of the QRS complexes. The aim of this analysis was determining whether human medicine algorithms for VT are applicable in veterinary medicine to differentiate wide QRS complex tachycardia in dogs. A retrospective analysis was performed on 11 dogs of both sexes and various breeds and age diagnosed with VT. The diagnosis was based on ambulatory ECG, further established based on the reaction to lidocaine or adenosine or an invasive electrophysiological study. Of the 11 tracings passed through the aVR algorithm, 10 met the VT criteria. The most common criterion was the Vi/Vt ratio (8 out of 11 tracings). Based on the VT score, seven out of eight dogs had a high probability of VT. Retrospective analysis of ECGs by aVR and VT score indicates that the applied algorithms may be useful in differentiating wide QRS complex tachycardia as a quick, easy, and non-invasive alternative to cardiac electrophysiology.
显示更多 [+] 显示较少 [-]Use of aVR and Jastrzębski algorithms in the classification of wide complex tachycardia in dogs – a preliminary study 全文
2018
Michałek Marcin | Frydrychowski Piotr | Adamowicz Jakub | Sławuta Agnieszka | Pasławska Urszula | Noszczyk-Nowak Agnieszka
Ventricular rhythm disturbances are a common pathology in human and veterinary medicine. In humans, the algorithmic approach is used to differentiate wide QRS complex tachycardia. The most commonly used are the aVR and Brugada algorithms as well as the ventricular tachycardia (VT) score developed by Jastrzębski and coworkers. In veterinary medicine, no such algorithms are available and the only parameter used to describe VT abnormalities is the duration of the QRS complexes. The aim of this analysis was determining whether human medicine algorithms for VT are applicable in veterinary medicine to differentiate wide QRS complex tachycardia in dogs.
显示更多 [+] 显示较少 [-]Identification and control of sources of Taenia solium infection – the attempts to eradicate the parasite 全文
2018
Samorek-Pieróg, Małgorzata | Karamon, Jacek | Cencek, Tomasz
Identification and control of sources of Taenia solium infection – the attempts to eradicate the parasite 全文
2018
Samorek-Pieróg, Małgorzata | Karamon, Jacek | Cencek, Tomasz
Taenia solium is a parasite causing porcine cysticercosis and human taeniosis and cysticercosis, parasitic zoonoses with a serious public health and economic influence. It has been globally ranked as the top foodborne parasite by the Food and Agriculture Organisation of the United Nations (FAO) and the World Health Organisation (WHO). This parasite is transmitted mainly in countryside regions where animals are free roaming, having access to human faeces, and infected pork is widely available. More developed countries eliminated cysticercosis; nonetheless, there are insufficient data about the current endemicity status of T. solium, due to increased human migration from endemic areas. Formally submitted statistics on cysticercosis in pigs are extremely inadequate. This is the result of not reporting all cases of the disease by some countries and lack of molecular verification during identification of the parasite. There is a need to develop diagnostic tests with increased sensitivity and specificity. The purpose of the present review is to summarise current knowledge about diagnostic and control methods concerning T. solium infection. The article does not address the diagnostics of human cysticercosis, since there is a distinct medical field which should be discussed separately. The paper focuses mainly on identifying the sources of T. solium infection, presenting the methods to detect and control porcine cysticercosis and taeniosis in humans.
显示更多 [+] 显示较少 [-]Identification and control of sources of Taenia solium infection – the attempts to eradicate the parasite 全文
2018
Samorek-Pieróg Małgorzata | Karamon Jacek | Cencek Tomasz
Taenia solium is a parasite causing porcine cysticercosis and human taeniosis and cysticercosis, parasitic zoonoses with a serious public health and economic influence. It has been globally ranked as the top foodborne parasite by the Food and Agriculture Organisation of the United Nations (FAO) and the World Health Organisation (WHO). This parasite is transmitted mainly in countryside regions where animals are free roaming, having access to human faeces, and infected pork is widely available. More developed countries eliminated cysticercosis; nonetheless, there are insufficient data about the current endemicity status of T. solium, due to increased human migration from endemic areas. Formally submitted statistics on cysticercosis in pigs are extremely inadequate. This is the result of not reporting all cases of the disease by some countries and lack of molecular verification during identification of the parasite. There is a need to develop diagnostic tests with increased sensitivity and specificity. The purpose of the present review is to summarise current knowledge about diagnostic and control methods concerning T. solium infection. The article does not address the diagnostics of human cysticercosis, since there is a distinct medical field which should be discussed separately. The paper focuses mainly on identifying the sources of T. solium infection, presenting the methods to detect and control porcine cysticercosis and taeniosis in humans.
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