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A K Siwicki - One of the best experts on this subject based on the ideXlab platform.

  • effects of Methisoprinol on innate immunity parameters in intensively reared european eel anguilla anguilla
    Israeli Journal of Aquaculture-bamidgeh, 2014
    Co-Authors: Edward Gląbski, B. Kazun, K Kazun, Stanislaw Robak, Agnieszka Lepa, A K Siwicki
    Abstract:

    Stress and chemotherapeutics can negatively affect the immune system of fish in intensive culture. Methods of prevention include immunonutrition. The present study examined the influence of dietary supplementation with the synthetic compound Methisoprinol on selected nonspecific immune parameters in intensively cultured juvenile European eel (Anguilla anguilla). The fish were fed a commercial pellet containing 0 (control) or 200 mg Methisoprinol/kg feed. After four weeks, the following immunological parameters were measured: respiratory burst activity and potential killing activity of the phagocytes, lymphocyte proliferation after stimulation by concanavaline A or lipopolisaccharide, and serum lysozyme activity, ceruloplasmin activity, total protein, and total immunoglobulin. Except for serum ceruloplasmin activity and total protein, all immune parameters were significantly higher (p<0.05) in the Methisoprinol-treated group than in the control, strongly suggesting that four weeks of feeding Methisoprinol to juvenile eels might improve innate immunity. The IJA appears exclusively as a peer-reviewed on-line open-access journal at http://www.siamb.org.il. To read papers free of charge, please register online at registration form. Sale of IJA papers is strictly forbidden.

  • impact of iridovirus and Methisoprinol on immunocompetent cells isolated from african catfish clarias gariepinus burchell under in vitro conditions
    Archives of Polish Fisheries, 2011
    Co-Authors: B. Kazun, A K Siwicki
    Abstract:

    Experiments to assess the effectiveness of the immunostimulator Methisoprinol (Polfa Grodzisk Pharmaceuticals, Poland) focused on its impact on the innate immune response. The impact of different doses of Methisoprinol on organ leukocytes isolated from the kidneys and spleens of African catfish that were subjected to or not subjected to the suppressive impact of iridovirus. The results indicate that the addition of Methisoprinol causes increased respiratory burst activity (RBA), potential killing activity (PKA), and proliferative activity of lymphocytes T and B in response to mitogens. Methisoprinol stimulates the mechanisms of cellular immunity that are linked to the activation of T lymphocytes, which can impact the antiviral activity of this preparation after its application in vivo.

  • in vitro assessment of the proliferative abilities of lymphocytes treated with respisure vaccine as well as immunomodulators lydium klp and Methisoprinol
    2010
    Co-Authors: W Szweda, A K Siwicki, A Plattsamoraj
    Abstract:

    There are no reports in the literature on the effect of vaccine antigen supplemented with immunomodulators on the activity of T and B lymphocytes. The aim of this study was to determine the proliferative activity of lymphocytes treated with a vaccine against mycoplasmal pneumonia of swine (MPS) and with selected immunomodulators in in vitro examinations. Five clinically healthy piglets at the age of 21 days were used in the experiment and blood for lymphocyte isolation was collected from them. Inactivated vaccine against MPS – Respisure (Pfizer) – at dilutions of 1 : 10, 1 : 100 and 1 : 1000 and immunomodulators – Methisoprinol (Polfa Grodzisk, Poland) and Lydium- KLP (Nika Health Products Ltd, USA), were also used. The test of mitogen-stimulated lymphocyte proliferation was performed according to the MTT method. T lymphocytes were stimulated with ConA, whereas B lymphocytes were stimulated with LPS. The experiment showed that combinations of immunomodulators: Lydium-KLP and Methisoprinol and the Respisure vaccine at dilutions of 1 : 100 and 1 : 1000 increase in vitro proliferative activity of ConA-stimulated T lymphocytes and LPS-stimulated B lymphocytes. A stronger proliferative response of both the lymphocyte types was observed for the combination of the Respisure vaccine and Methisoprinol.

  • the influence of Methisoprinol on the spleen phagocyte and blood lymphocyte activity in rats in vitro study
    2009
    Co-Authors: A K Siwicki, Leszek Jung, Roman Wojcik
    Abstract:

    The application of immunostimulators in human and veterinary medicine offers a wide of attractive method for inducing or modulating protection against infection diseases. In the present study we determined the in vitro influence of different concentrations of synthetic product Methisoprinol on the proliferative response of blood lymphocytes stimulated by concanavaline A (ConA) and lipopolisaccharide (LPS) in rats. Also the metabolic ability and potential killing activity of spleen phagocytes were examined. For this study 10 adult male and 10 adult female rats were used. Peripheral blood was obtained by venous puncture (Vacutainer set). The proliferative ability of the blood lymphocytes stimulated by mitogens was determined by MTT assay. The respiratory burst activity (RBA) and potential killing activity (PKA) were determined by spectrophotometric assay. In each experiment, the concentrations of Methisoprinol used in the RPMI 1640 medium were 0, 0.5, 1, 5, 10, 25 and 50 µg methisorinol/ml of medium. The present study shows that a concentrations between 5 to 50 µg/ml of Methisoprinol statistically significantly (p < 0.05) increase the mitogens-induced proliferation rate of rats lymphocytes T and B and we have not observed statistically significant difference between male and female. The analysis of the results showed that the Methisoprinol increased the metabolic ability (RBA) and potential killing activity (PKA) of spleen phagocytes at concentrations between 5 and 50 µg/ml, compared to the control. The highest RBA and PKA were observed at concentrations between 10 to 50 µg/ml and also we have not observed statistically significant difference between male and female animals.

  • immunomodulating effect of Methisoprinol on the pronephros macrophage and lymphocyte activity after suppression induced by infectious haematopoietic necrosis virus ihnv in rainbow trout oncorhynchus mykiss
    Acta Veterinaria Brno, 2008
    Co-Authors: A K Siwicki, B. Kazun, J Malaczewska, Roman Wojcik
    Abstract:

    Siwicki A. K, J. Malaczewska, B. Kazun, R. Wojcik: Immunomodulating Effect of Methisoprinol on the Pronephros Macrophage and Lymphocyte Activity after Suppression Induced by Infectious Haematopoietic Necrosis Virus (IHNV) in Rainbow Trout (Oncorhynchus mykiss). Acta Vet. Brno 2008, 77: 631-635. The purpose of this in vitro study was to determine the influence of Methisoprinol on the activity of pronephros macrophages and lymphocytes after suppression induced by infectious haematopoietic necrosis virus (IHNV). For this study IHNV-free rainbow trout were used. Pronephros from 20 fish were removed and single leukocyte suspensions were separated. The IHNV significantly (P < 0.05) decreased the respiratory burst activity and potential killing activity of pronephros macrophages and proliferative response of lymphocytes stimulated by mitogens ConA and LPS. The results of our in vitro study showed that Methisoprinol at a concentration of 50 mg/ml modulated (restored) the metabolic and potential killing activity of macrophages and proliferative response of lymphocytes suppressed by IHNV. In vitro study, phagocyte activity, lymphocyte proliferation Rhabdoviruses constitute one of the largest groups of viruses isolated from fish, and are mostly associated with epizootics and heavy losses in intensive fish farming. Infectious haematopoietic necrosis (IHN) is the most important viral disease and produces high losses in rainbow trout and other salmon species farmed in North America and in several European countries. Infectious haematopoietic necrosis virus (IHNV) is a member of Rhabdoviridae and the type species of the genus Novirhabdovirus. Like other mononegavirales, it has a single molecule of linear, negative-sense ssRNA genome (Kurath et al. 1997). The multiplication of virus takes place in endothelial cells of blood capillaries leading to haemorrhages in haematopoietic tissues and nephron cells (Amend and Smith 1974). The transmission of IHNV takes place horizontally, vertically, and by biological vectors such as fish parasites. Acutely infected fish release the virus by external mucus, faeces and urine. Carriers shed the agent via sexual products (Kim et al. 1999). The protection against viral diseases by specific vaccines against IHN is being developed for the last few years (Corbeil et al. 2000; Lorenzen et al. 2002; Prost 2003) and some successful vaccines have been developed (Purcell et al. 2006; Miller et al. 2007). Rodriguez Saint-Jean and Perez-Prieto (2007) examined the ability of several fish viruses to induce protection against homologous or heterologous viruses in single or double infections, and assessed whether such protection is correlated with innate immunity or expression of the Mx gene. The results of this study indicate that activation of the immune response could explain the interference and loss of IHNV in the IPNV-IHNV co-infections. In fact, the DNA vaccines against IHN virus only have a scientific aspect and the application of these vaccines in rainbow trout culture is ACTA VET. BRNO 2008, 77: 631-635; doi:10.2754/avb200877040631 Address for correspondence: Andrzej K. Siwicki Zabieniec near Warsaw 05-500 Piaseczno POLAND E-mail: aksiw@infish.com.pl http://www.vfu.cz/acta-vet/actavet.htm limited. However, for an ideal preventive approach, specific drugs should be developed to inhibit selectively virus replication or to stimulate the antiviral protection. The use of natural and synthetic immunomodulators in fish offers a wide range of attractive methods for inducing and building up protection against viral diseases and is a promising new development in aquaculture (Anderson 1992; Siwicki et al. 1998). Several promising drugs and biological response modifiers such as carbohydrates and other synthetics have been tested on fish immunocompetence cells in vitro. A dosedependent immunomodulatory effect of levamisole, nitrogranulogen, lysozyme dimer and HMB were observed on macrophage activity, proliferative response of lymphocytes and antibody secreting cells in rainbow trout and other fish species (Siwicki et al. 2003; Terech-Majewska et al. 2004; Siwicki et al. 2006). Methisoprinol is a synthetic compound formed from the p-acetamidobenzoate salt of N-N-dimethylamino-2-propanol and inosine in a 3 : 1 molar ratio. Methisoprinol presents low toxicity and has been shown to act in vitro by inhibiting the replication of salmonid rhabdoviruses (Siwicki et al. 2002). It exerts antiviral and antitumour activities in vitro and in vivo, which are secondary to the immunomodulating influence on non-specific cellular and humoral defence mechanisms and protection against viral diseases (Delogu et al. 1982; Fudenberg and Whitten 1984; Siwicki et al. 2003). The aim of the present study was to determine the in vitro influence of Methisoprinol on the pronephros macrophage and lymphocyte activity after suppression induced by infectious haematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss). Materials and Methods In this in vitro study 20 healthy (IHNV-free) rainbow trout (Oncorhynchus mykiss) with a mean weight of 50 g were used. The fish were held in a 500 l tank in 14 oC spring water and fed twice daily with commercial pellets. The pronephros were removed from fish aseptically and single leukocyte suspensions were separated on Histopaque-1077 (Sigma, USA) gradient or Gradisol (Polfa, Poland), according to the method presented by Siwicki et al. (1996). Viable cells from the pronephros were counted with 0.1% of trypan blue staining after washing three times in RPMI 1640 medium, and 90–94% of cell vitality were ascertained. The Laboratoire Departemental d’Analyses France (LDA 39) IHNV isolate was used in this in vitro study and quantified by plaque assay using epithelioma papulosum cyprini (EPC) cells incubated at 14 °C for 72 h. Methisoprinol (Polfa Grodzisk, Poland) was used at a concentration of 50 mg/ml of RMPI-1640 medium (Sigma). The spectrophotometric assay presented by Siwicki et al. (1996) was used to study the respiratory burst activity of pronephros macrophages stimulated by phorbol myristate acetate (PMA, Sigma). One hundred ml of cell suspension (1 × 106 in RPMI-1640) were added to 96-well culture plates (Nunclon, Denmark) and incubated for 2 h at 22 oC with 50 ml of Methisoprinol or 50 ml of 1 × 107 plaque-forming units/ml RPMI-1640 of IHN virus or with 50 ml of IHN virus + 50 ml of Methisoprinol. The control group comprised cells only stimulated by PMA. The potential killing activity of pronephros macrophages was determined with a microcolorimetric method presented by Anderson and Siwicki (1994). The 0.2% nitroblue tetrazolium (NBT, Sigma, USA) in PBS solution containing live 1 × 107 Aeromonas salmonicida cells was used for stimulation of pronephros macrophages. The cells (100 ml) were added to 96-well culture plates (Nunclon, Denmark) and incubated for 30 min at 22 oC with 50 ml of Methisoprinol or 1 × 107 plaque-forming units/ml RPMI-1640 of IHN virus or with 50 ml of IHN virus + 50 ml of Methisoprinol. The control group comprised cells only stimulated by 0.2% NBT solution containing live bacteria A. salmonicida. The proliferative response of pronephros lymphocytes was determined by the MTT [3-(4,5-dimethyl thiazol2-yl)-2,5-diphenyl tetrazolium bromide] assay described by Mosmann (1983), as modified for use with fish lymphocytes by Siwicki et al. (1996). Briefly, lymphocytes were isolated from pronephros and distributed (100 ml) into 96-well culture plates (Nunclon, Denmark) at 5 × 106 cell/ml of RPMI-1640. The mitogen concanavalin A (ConA, Sigma) at a concentration of 64 mg/ml or lipopolysaccharide (LPS, Sigma) at a concentration of 160 mg/ml was added (20 ml) into each well. The cells were incubated for 72 h with 50 ml of Methisoprinol or 50 ml of IHN virus (1 × 107 pfu/ml) or with 50 ml of IHN virus + 50 ml of Methisoprinol. The control group comprised cells only stimulated by mitogens: Con A or LPS. The results were verified statistically by one-way ANOVA analysis of variance (GraphPad Prism software 632

L. Krakowski - One of the best experts on this subject based on the ideXlab platform.

  • Selected leukocyte subpopulations in peripheral blood and uterine washings in cows before and after intrauterine administration of cefapirin and Methisoprinol.
    Animal science journal = Nihon chikusan Gakkaiho, 2019
    Co-Authors: Piotr Brodzki, Urszula Lisiecka, Adam Brodzki, L. Krakowski, Marek Szczubiał, Roman Dąbrowski, Andrzej Junkuszew, M. Bochniarz
    Abstract:

    The aim of the study was to evaluate the selected lymphocyte subpopulations TCD4, TCD8, BCD21, BCD25, CD18, CD11b, and MHC II in blood and uterine flush of cows with endometritis, before and after intrauterine (i.u.) administration of cefapirin and Methisoprinol. The research was carried out on 28 cows with clinical endometritis. Animals were divided into four groups, each composed of seven cows, depending on the i.u. preparation used: Group A, cefapirin; Group B, Methisoprinol; Group C, cefapirin and Methisoprinol simultaneously; and a control group-without medication. The study was performed using flow cytometry method. Summarizing the results of the research, i.u. infusion of cefapirin caused a weakening of the effector phase of the local uterine immune response; however, it enhanced leukocyte chemotaxis and antigen presentation. After i.u. administration of Methisoprinol, the stimulation of specific uterine immunity mechanisms was mainly observed. The use of both mentioned preparations showed the strengthening of specific uterine immunological mechanisms presumably caused by Methisoprinol, despite the inhibitory effect of the antibiotic. Intrauterine use of immunostimulatory substances can improve the effectiveness of the endometritis treatment in cows by improving specific local mechanisms of uterine immunity. As a consequence, it may enhance the effector function of immune competent cells and finally eliminate inflammation.

  • phagocytic and oxidative burst activity of phagocytic cells in peripheral blood and uterine washings in cows with clinical endometritis before and after intrauterine use of cephapirin and Methisoprinol
    Animal Science Journal, 2018
    Co-Authors: Piotr Brodzki, Urszula Lisiecka, Adam Brodzki, Renata Pyzłukasik, L. Krakowski
    Abstract:

    The aim of the study was to evaluate phagocytic and killing activity of phagocytic cells in blood and uterine flush of cows with endometritis before and after intrauterine (i.u.) administration of cephapirin and Methisoprinol. The research was carried out on 28 cows with clinical endometritis. Animals were divided into four groups, each composed of seven cows, depending on the i.u. treatment used: Group A-cephapirin; Group B-Methisoprinol; Group C-cephapirin and Methisoprinol at the same time; and a control group-without medication. Using flow cytometry technique, the phagocytic activity of granulocytes and monocytes was identified, as well as the oxidative burst activity of neutrophils in the peripheral blood and uterine washings. Summarizing the results of the research, i.u. infusion of cephapirin caused a reduction in the phagocytic and killing activity of phagocytes. The i.u. use of Methisoprinol increased phagocytic and killing activity of phagocytes in the uterus. Administering both listed substances simultaneously showed a decrease in phagocytosis, presumably due to the dominating inhibitor effect of the antibiotic. However, also an increase of mean fluorescence intensity was observed, presumably caused by the Methisoprinol. Intrauterine use of immunostimulatory substances, can improve the effectiveness of the treatment of endometritis in cows.

B. Kazun - One of the best experts on this subject based on the ideXlab platform.

  • effects of Methisoprinol on innate immunity parameters in intensively reared european eel anguilla anguilla
    Israeli Journal of Aquaculture-bamidgeh, 2014
    Co-Authors: Edward Gląbski, B. Kazun, K Kazun, Stanislaw Robak, Agnieszka Lepa, A K Siwicki
    Abstract:

    Stress and chemotherapeutics can negatively affect the immune system of fish in intensive culture. Methods of prevention include immunonutrition. The present study examined the influence of dietary supplementation with the synthetic compound Methisoprinol on selected nonspecific immune parameters in intensively cultured juvenile European eel (Anguilla anguilla). The fish were fed a commercial pellet containing 0 (control) or 200 mg Methisoprinol/kg feed. After four weeks, the following immunological parameters were measured: respiratory burst activity and potential killing activity of the phagocytes, lymphocyte proliferation after stimulation by concanavaline A or lipopolisaccharide, and serum lysozyme activity, ceruloplasmin activity, total protein, and total immunoglobulin. Except for serum ceruloplasmin activity and total protein, all immune parameters were significantly higher (p<0.05) in the Methisoprinol-treated group than in the control, strongly suggesting that four weeks of feeding Methisoprinol to juvenile eels might improve innate immunity. The IJA appears exclusively as a peer-reviewed on-line open-access journal at http://www.siamb.org.il. To read papers free of charge, please register online at registration form. Sale of IJA papers is strictly forbidden.

  • Impact of Bioimmuno with Methisoprinol on non-specific cellular and humoral defense mechanisms and resistance of African catfish (Clarias gariepinus) to experimental infection with iridovirus
    Archives of Polish Fisheries, 2013
    Co-Authors: B. Kazun, Andrzej K. Siwicki
    Abstract:

    The aim of the experiment was to determine the impact Bioimmuno had on the non-specific cellular and humoral defense mechanisms and resistance to experimental infection with iridovirus of African catfish, Clarias gariepinus (Burchell). The experiment was performed on clinically healthy African catfish weighing from 30 to 200 g. The fish were infected experimentally with iridovirus 59.90 at a dose of 10 4 TCID50 ml -1 . Bioimmuno administered at a dose of 1 kg per 50 kg of feed most effectively stimulated the responses of non-specific cellular and humoral defense mechanisms, corrected delayed immunological reactions induced by iridovirus infection, and limited mortality caused by experimental viral infection of 5-45%.

  • impact of iridovirus and Methisoprinol on immunocompetent cells isolated from african catfish clarias gariepinus burchell under in vitro conditions
    Archives of Polish Fisheries, 2011
    Co-Authors: B. Kazun, A K Siwicki
    Abstract:

    Experiments to assess the effectiveness of the immunostimulator Methisoprinol (Polfa Grodzisk Pharmaceuticals, Poland) focused on its impact on the innate immune response. The impact of different doses of Methisoprinol on organ leukocytes isolated from the kidneys and spleens of African catfish that were subjected to or not subjected to the suppressive impact of iridovirus. The results indicate that the addition of Methisoprinol causes increased respiratory burst activity (RBA), potential killing activity (PKA), and proliferative activity of lymphocytes T and B in response to mitogens. Methisoprinol stimulates the mechanisms of cellular immunity that are linked to the activation of T lymphocytes, which can impact the antiviral activity of this preparation after its application in vivo.

  • immunomodulating effect of Methisoprinol on the pronephros macrophage and lymphocyte activity after suppression induced by infectious haematopoietic necrosis virus ihnv in rainbow trout oncorhynchus mykiss
    Acta Veterinaria Brno, 2008
    Co-Authors: A K Siwicki, B. Kazun, J Malaczewska, Roman Wojcik
    Abstract:

    Siwicki A. K, J. Malaczewska, B. Kazun, R. Wojcik: Immunomodulating Effect of Methisoprinol on the Pronephros Macrophage and Lymphocyte Activity after Suppression Induced by Infectious Haematopoietic Necrosis Virus (IHNV) in Rainbow Trout (Oncorhynchus mykiss). Acta Vet. Brno 2008, 77: 631-635. The purpose of this in vitro study was to determine the influence of Methisoprinol on the activity of pronephros macrophages and lymphocytes after suppression induced by infectious haematopoietic necrosis virus (IHNV). For this study IHNV-free rainbow trout were used. Pronephros from 20 fish were removed and single leukocyte suspensions were separated. The IHNV significantly (P < 0.05) decreased the respiratory burst activity and potential killing activity of pronephros macrophages and proliferative response of lymphocytes stimulated by mitogens ConA and LPS. The results of our in vitro study showed that Methisoprinol at a concentration of 50 mg/ml modulated (restored) the metabolic and potential killing activity of macrophages and proliferative response of lymphocytes suppressed by IHNV. In vitro study, phagocyte activity, lymphocyte proliferation Rhabdoviruses constitute one of the largest groups of viruses isolated from fish, and are mostly associated with epizootics and heavy losses in intensive fish farming. Infectious haematopoietic necrosis (IHN) is the most important viral disease and produces high losses in rainbow trout and other salmon species farmed in North America and in several European countries. Infectious haematopoietic necrosis virus (IHNV) is a member of Rhabdoviridae and the type species of the genus Novirhabdovirus. Like other mononegavirales, it has a single molecule of linear, negative-sense ssRNA genome (Kurath et al. 1997). The multiplication of virus takes place in endothelial cells of blood capillaries leading to haemorrhages in haematopoietic tissues and nephron cells (Amend and Smith 1974). The transmission of IHNV takes place horizontally, vertically, and by biological vectors such as fish parasites. Acutely infected fish release the virus by external mucus, faeces and urine. Carriers shed the agent via sexual products (Kim et al. 1999). The protection against viral diseases by specific vaccines against IHN is being developed for the last few years (Corbeil et al. 2000; Lorenzen et al. 2002; Prost 2003) and some successful vaccines have been developed (Purcell et al. 2006; Miller et al. 2007). Rodriguez Saint-Jean and Perez-Prieto (2007) examined the ability of several fish viruses to induce protection against homologous or heterologous viruses in single or double infections, and assessed whether such protection is correlated with innate immunity or expression of the Mx gene. The results of this study indicate that activation of the immune response could explain the interference and loss of IHNV in the IPNV-IHNV co-infections. In fact, the DNA vaccines against IHN virus only have a scientific aspect and the application of these vaccines in rainbow trout culture is ACTA VET. BRNO 2008, 77: 631-635; doi:10.2754/avb200877040631 Address for correspondence: Andrzej K. Siwicki Zabieniec near Warsaw 05-500 Piaseczno POLAND E-mail: aksiw@infish.com.pl http://www.vfu.cz/acta-vet/actavet.htm limited. However, for an ideal preventive approach, specific drugs should be developed to inhibit selectively virus replication or to stimulate the antiviral protection. The use of natural and synthetic immunomodulators in fish offers a wide range of attractive methods for inducing and building up protection against viral diseases and is a promising new development in aquaculture (Anderson 1992; Siwicki et al. 1998). Several promising drugs and biological response modifiers such as carbohydrates and other synthetics have been tested on fish immunocompetence cells in vitro. A dosedependent immunomodulatory effect of levamisole, nitrogranulogen, lysozyme dimer and HMB were observed on macrophage activity, proliferative response of lymphocytes and antibody secreting cells in rainbow trout and other fish species (Siwicki et al. 2003; Terech-Majewska et al. 2004; Siwicki et al. 2006). Methisoprinol is a synthetic compound formed from the p-acetamidobenzoate salt of N-N-dimethylamino-2-propanol and inosine in a 3 : 1 molar ratio. Methisoprinol presents low toxicity and has been shown to act in vitro by inhibiting the replication of salmonid rhabdoviruses (Siwicki et al. 2002). It exerts antiviral and antitumour activities in vitro and in vivo, which are secondary to the immunomodulating influence on non-specific cellular and humoral defence mechanisms and protection against viral diseases (Delogu et al. 1982; Fudenberg and Whitten 1984; Siwicki et al. 2003). The aim of the present study was to determine the in vitro influence of Methisoprinol on the pronephros macrophage and lymphocyte activity after suppression induced by infectious haematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss). Materials and Methods In this in vitro study 20 healthy (IHNV-free) rainbow trout (Oncorhynchus mykiss) with a mean weight of 50 g were used. The fish were held in a 500 l tank in 14 oC spring water and fed twice daily with commercial pellets. The pronephros were removed from fish aseptically and single leukocyte suspensions were separated on Histopaque-1077 (Sigma, USA) gradient or Gradisol (Polfa, Poland), according to the method presented by Siwicki et al. (1996). Viable cells from the pronephros were counted with 0.1% of trypan blue staining after washing three times in RPMI 1640 medium, and 90–94% of cell vitality were ascertained. The Laboratoire Departemental d’Analyses France (LDA 39) IHNV isolate was used in this in vitro study and quantified by plaque assay using epithelioma papulosum cyprini (EPC) cells incubated at 14 °C for 72 h. Methisoprinol (Polfa Grodzisk, Poland) was used at a concentration of 50 mg/ml of RMPI-1640 medium (Sigma). The spectrophotometric assay presented by Siwicki et al. (1996) was used to study the respiratory burst activity of pronephros macrophages stimulated by phorbol myristate acetate (PMA, Sigma). One hundred ml of cell suspension (1 × 106 in RPMI-1640) were added to 96-well culture plates (Nunclon, Denmark) and incubated for 2 h at 22 oC with 50 ml of Methisoprinol or 50 ml of 1 × 107 plaque-forming units/ml RPMI-1640 of IHN virus or with 50 ml of IHN virus + 50 ml of Methisoprinol. The control group comprised cells only stimulated by PMA. The potential killing activity of pronephros macrophages was determined with a microcolorimetric method presented by Anderson and Siwicki (1994). The 0.2% nitroblue tetrazolium (NBT, Sigma, USA) in PBS solution containing live 1 × 107 Aeromonas salmonicida cells was used for stimulation of pronephros macrophages. The cells (100 ml) were added to 96-well culture plates (Nunclon, Denmark) and incubated for 30 min at 22 oC with 50 ml of Methisoprinol or 1 × 107 plaque-forming units/ml RPMI-1640 of IHN virus or with 50 ml of IHN virus + 50 ml of Methisoprinol. The control group comprised cells only stimulated by 0.2% NBT solution containing live bacteria A. salmonicida. The proliferative response of pronephros lymphocytes was determined by the MTT [3-(4,5-dimethyl thiazol2-yl)-2,5-diphenyl tetrazolium bromide] assay described by Mosmann (1983), as modified for use with fish lymphocytes by Siwicki et al. (1996). Briefly, lymphocytes were isolated from pronephros and distributed (100 ml) into 96-well culture plates (Nunclon, Denmark) at 5 × 106 cell/ml of RPMI-1640. The mitogen concanavalin A (ConA, Sigma) at a concentration of 64 mg/ml or lipopolysaccharide (LPS, Sigma) at a concentration of 160 mg/ml was added (20 ml) into each well. The cells were incubated for 72 h with 50 ml of Methisoprinol or 50 ml of IHN virus (1 × 107 pfu/ml) or with 50 ml of IHN virus + 50 ml of Methisoprinol. The control group comprised cells only stimulated by mitogens: Con A or LPS. The results were verified statistically by one-way ANOVA analysis of variance (GraphPad Prism software 632

  • influence of Methisoprinol on the replication of rhabdoviruses isolated from carp cyprinus carpio and catfish ictalurus melas in vitro study
    Polish Journal of Veterinary Sciences, 2003
    Co-Authors: A K Siwicki, B. Kazun, F Pozet, M Morand, S Trapkowska, J Malaczewska
    Abstract:

    Rhabdoviruses constitute one of the most pathogenic viruses isolated from rainbow trout and carp culture. Several viruses were also isolated from other species of fish. These viruses are mostly associated with epizootics and heavy losses. Spring viraemia of carp virus (SVCV) and pike fry rhabdovirus (PFRV) have been the most extensively studied, due to their significant economic impact. Significant progress has been made towards controlling the major bacterial fish diseases using vaccines, but this approach has not yet been successful in preventing viral diseases in fish culture. However, for an effective therapeutic approach, specific drugs should be developed to selectively inhibit virus replication and/or stimulate antiviral protection. In this investigation we examined the in vitro influence of Methisoprinol on the SVCV and virus isolated from catfish (Ictalurus melas) replication by measuring their RNA synthesis. The viruses were propagated in EPC cells and cell cultures containing Methisoprinol were followed by infection with SVCV or catfish rhabdovirus suspension containing 10(7) TCID50/ml. Methisoprinol (Polfa, Poland) at concentrations of 0, 100, 200, 300, 400 and 500 microg/ml of medium (Glasgow MEM) was used in this study. The results of this study show the strong inhibition of incorporation (cpm) of [3H]-uridine into SVCV and catfish rhabdovirus RNA in cell culture exposed to Methisoprinol at various concentrations. The highest percent of inhibition of viral RNA at 72 h after infection with two rhabdoviruses were observed in doses of 400 and 500 microg/ml of Methisoprinol in medium. The results of this in vitro study showed that Methisoprinol inhibits the rhabdoviruses isolated from carp and catfish.

Andrzej K. Siwicki - One of the best experts on this subject based on the ideXlab platform.

Piotr Brodzki - One of the best experts on this subject based on the ideXlab platform.

  • Selected leukocyte subpopulations in peripheral blood and uterine washings in cows before and after intrauterine administration of cefapirin and Methisoprinol.
    Animal science journal = Nihon chikusan Gakkaiho, 2019
    Co-Authors: Piotr Brodzki, Urszula Lisiecka, Adam Brodzki, L. Krakowski, Marek Szczubiał, Roman Dąbrowski, Andrzej Junkuszew, M. Bochniarz
    Abstract:

    The aim of the study was to evaluate the selected lymphocyte subpopulations TCD4, TCD8, BCD21, BCD25, CD18, CD11b, and MHC II in blood and uterine flush of cows with endometritis, before and after intrauterine (i.u.) administration of cefapirin and Methisoprinol. The research was carried out on 28 cows with clinical endometritis. Animals were divided into four groups, each composed of seven cows, depending on the i.u. preparation used: Group A, cefapirin; Group B, Methisoprinol; Group C, cefapirin and Methisoprinol simultaneously; and a control group-without medication. The study was performed using flow cytometry method. Summarizing the results of the research, i.u. infusion of cefapirin caused a weakening of the effector phase of the local uterine immune response; however, it enhanced leukocyte chemotaxis and antigen presentation. After i.u. administration of Methisoprinol, the stimulation of specific uterine immunity mechanisms was mainly observed. The use of both mentioned preparations showed the strengthening of specific uterine immunological mechanisms presumably caused by Methisoprinol, despite the inhibitory effect of the antibiotic. Intrauterine use of immunostimulatory substances can improve the effectiveness of the endometritis treatment in cows by improving specific local mechanisms of uterine immunity. As a consequence, it may enhance the effector function of immune competent cells and finally eliminate inflammation.

  • phagocytic and oxidative burst activity of phagocytic cells in peripheral blood and uterine washings in cows with clinical endometritis before and after intrauterine use of cephapirin and Methisoprinol
    Animal Science Journal, 2018
    Co-Authors: Piotr Brodzki, Urszula Lisiecka, Adam Brodzki, Renata Pyzłukasik, L. Krakowski
    Abstract:

    The aim of the study was to evaluate phagocytic and killing activity of phagocytic cells in blood and uterine flush of cows with endometritis before and after intrauterine (i.u.) administration of cephapirin and Methisoprinol. The research was carried out on 28 cows with clinical endometritis. Animals were divided into four groups, each composed of seven cows, depending on the i.u. treatment used: Group A-cephapirin; Group B-Methisoprinol; Group C-cephapirin and Methisoprinol at the same time; and a control group-without medication. Using flow cytometry technique, the phagocytic activity of granulocytes and monocytes was identified, as well as the oxidative burst activity of neutrophils in the peripheral blood and uterine washings. Summarizing the results of the research, i.u. infusion of cephapirin caused a reduction in the phagocytic and killing activity of phagocytes. The i.u. use of Methisoprinol increased phagocytic and killing activity of phagocytes in the uterus. Administering both listed substances simultaneously showed a decrease in phagocytosis, presumably due to the dominating inhibitor effect of the antibiotic. However, also an increase of mean fluorescence intensity was observed, presumably caused by the Methisoprinol. Intrauterine use of immunostimulatory substances, can improve the effectiveness of the treatment of endometritis in cows.