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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.
Aim of the study: In view of the frequent occurrence of immunosuppression in pigeons, which is a consequence of viral infections, studies with the use of a synthetic immunomodulator (methisoprinol) were undertaken to evaluate its impact on the course of an experimental infection with PPMV-1. The aim of the study was to determine the usefulness of methisoprinol for the treatment or prophylaxis of viral infections in pigeons. Materials and Methods: Three groups of 5-week-old pigeons, with 15 birds in each, were used in the experiment. Before the experiment, the birds were tested for the presence of antibodies specific to paramyxovirus, and their sensitivity to PPMV-1 infection was evaluated. The virus had been cultured earlier on SPF chicken embryos of 9-12 days. The pigeons from all three groups were infected intravenously with aparamyxovirus suspension (strain APMV-1/ pigeon/Poland/AR3/95 obtained from the Veterinary Institute in Puławy) at a concentration of LD50 10-7 in 0.1 ml at a dose of 0.1 ml/pigeon. The birds in the experimental groups (B1 and B2) were immunomodulated with methisoprinol administered at a dose of 200 mg/1 kg of body weight. The immunomodulator was administered by intramuscular injection for 3 successive days before (group B1) or three successive days after (group B2) the experimental infection. Pigeons in group K (used as a control) were given water by intramuscular injection for 3 days before and 3 days after infection. On days 4, 8, and 12 after infection, 5 birds from each group were euthanized, and sections of internal organs (lung, kidney, brain), as well as cloacal swabs, were collected to detect viral RNA by the RT-PCR method. Results and discussion: Symptoms were recorded from the first day after infection. Neurological symptoms occurred in birds from all groups: in 100% of pigeons from groups B1 and K, and in 80% of pigeons from group B2. Deaths of birds occurred from day 5 after infection in group B1. In the other groups, deaths were observed from day 6 after infection. The total mortality of the infected birds ranged from 70% (group B2) to 100% (groups B1 and K). The resolution of symptoms was observed from day 6 after infection in pigeons from group B2. During molecular examination, it was noted that the highest number of positive samples (presence of PPMV-1) on each day of the investigation was obtained from brain samples and cloacal swabs. The highest number of positive results in kidney samples was obtained from groups B1 and K on day 4 after infection. On the successive days of the investigation the percentage of positive samples increased to 100 in birds from all groups except group B2. Based On the basis of the results of the present study, it can be concluded that methisoprinol, used at a dose of 200 mg/kg of body weight after infection, has antiviral activity, manifested by a slower development of paramyxovirosis in pigeons infected intravenously with PPMV-1. Therefore, the administration of methisoprinol to naturally infected and diseased birds may be useful in the treatment of viral diseases.
The aim of the research was to determine the efficiency of in ovo immunisation of turkeys against the haemorrhagic enteritis virus, while simultaneously applying a synthetic immunomodulator - methisoprinol - by the same route of administration. Dindoral SPF vaccine, in a dose of 0.1 ml of the solution prepared ex tempore after dissolving the vaccine in 100 ml of water for injection, and methisoprinol in a dose of 5 mg per egg, were administered in ovo on the 26th d of incubation. The control groups consisted of turkeys hatched from eggs into which only the methisoprinol or the vaccine was administered, as well as birds hatched from eggs that were not interfered with. The susceptibility of turkeys to HE virus infection was determined on the basis of the presence of HE antibodies in serum, the evaluation of a splenic index, and attempts to register the virus presence in the spleen 120 h after the control infections. The research proved the effectiveness of immunising turkeys against the haemorrhagic enteritis virus by administering Dindoral SPF vaccine in ovo. It was also demonstrated that the simultaneous application of methisoprinol, showing an antiviral effect, and the vaccine inhibited the development of post vaccinal immunity against this virus.
The application of immunostimulators in dogs offers a novel approach to induce or modulate the mechanisms of protection against infectious diseases. In the present studies we evaluated in vitro influence of different concentrations of synthetic product - methisoprinol on the proliferative response of blood lymphocytes stimulated by pokeweed mitogen (PWM), concanavaline A and lipopolisaccharide (LPS) in dogs. The concentrations of methisoprinol between 1 to 50 μg/ml increased the mitogen-induced proliferation rate of canine T and B lymphocytes. Enhanced response to mitogens was observed both in 1-2 and 6-8 years old dogs, but the best results were observed in older animals. The results suggested that methisoprinol demonstrated a stronger effect on the lymphocytes isolated from older animals with the lower proliferative responses.
As a consequence of frequent immunosuppression in pigeons, with resultant decreased post-vaccination immunity and deteriorated health of the birds, a study was taken up in order to determine the effect of three immunomodulators (β-glucans, metisoprinol and levamisole) on the percentage of CD4⁺ and CD8⁺ T lymphocytes in peripheral blood and spleen, and the titre of anti-NDV antibodies in the serum of pigeons in four groups (A, B, C, D), 20 birds in each. The birds in each group were immunized against paramyxovirosis in week 6 and 9 of their lives, and water for injection was given intramuscularly 1 day before each injection (group A control), metisoprinol was given intramuscularly for 3 days at a dose of 300 mg/kg of body weight (group B), levamisole was given as a 7.5% solution of levamisole hydrochloride at a dose of 2 mg/kg of body weight intramuscularly, 1 day (group C), or β-glucans were given 10 days before vaccination per os at a dose of 5 mg/kg of body weight (group D). The immunological examinations were carried out by flow cytometry and the ELISA test. The results indicate that levamisole and β-glucans at the doses used in the study stimulate an increase in the anti-NDV antibody titre and the percentage of CD4⁺ T lymphocyte subpopulation in peripheral blood (levamisole) along with an increase in the percentage of CD8⁺ T lymphocytes in the spleen of pigeons vaccinated against paramyxovirosis. The absence of such an effect following the administration of metisoprinol at a dose of 300 mg/kg of body weight for 3 days may have resulted from an excessively high dose.
A study was undertaken to determine the effect of a synthetic immunomodulator, i.e. methisop- rinol applied in ovo, upon the hatchability of turkey poults under conditions of a standard hatchery as well as on their health status evaluated based on analyses of selected biochemical indices in their blood serum. Experiments were conducted on 5 groups of BUT 9 turkeys at the age of 5 days (35 birds in each group) hatched from eggs to which methisoprinol (VetAgro, Lublin, Poland) was applied in ovo at a dose of 5 mg (group I), 10 mg (group II) or 20 mg per egg (group III) on the 26th day of incubation. Turkeys hatched from eggs to which a physiological solution of NaCl was applied on the same day at a dose of 0.1 ml per egg (group IV) as well as those hatched from eggs without in ovo injection (group V) served as controls. Five hundreds eggs were used in each group. Hatchability was evaluated based on the number of hatched poults in respect of the number of eggs with live embryos transferred from the setting compartment to the hatching compartment, that were subjected to in ovo administration of the preparations according to the experimental design. Blood serum of the 5-day-old turkey poults was analyzed for activities of AST, ALP, LDH-L, CK, lysozyme and cerulop- lasmine as well as for total protein and albumin contents. Analyses were also conducted for the immune system organ index - percentage contribution of organs of the immune system (spleen, thymus and the bursa of Fabricius) in the body weight of turkeys. The study demonstrated that methisoprinol administered to turkey embryos in ovo on day 26 of incubation at doses of 5, 10 or 20 mg per embryo did not induce any disturbances in the hatching process or affect its final result. In addition, it was shown not to exert any negative effect on the health status of the reared turkey poults.
The study was aimed on determining the clinical protective value of Lydium-KLP and Methisoprinol in the prevention of respiratory system diseases, as well as their effect on the health status of pigs in large-scale commercial breeding and on the improvement of in the effectiveness of specific immunoprophylaxis of mycoplasmal pneumonia of in swine (MPS). Piglets at the age of 7 d were divided into eight groups of 25-30 piglets each. On days 7 and 21 of life they were immunised with Respisure vaccine (group I) or administrated Lydium-KLP or Methisoprinol (groups IV and VII). In other groups, Lydium-KLP or Methisoprinol were given simultaneously (groups II and V) or 48 h before immunisation with Respisure (groups III and VI). Control piglets were administrated PBS (group C). Clinical observations as well as post-slaughter and animal husbandry analyses indicated that the simultaneous administration of Lydium-KLP or Methisoprinol with Respisure gives better results in comparison to immunisation- only with Respisure. The statistically-significant differences were found in the mean percentage of meatiness between groups receiving Methisoprinol with or 48 h before Respisure and control group, as well as in the point evaluation of lesions in the lungs between groups receiving Lydium-KLP with Respisure and Methisoprinol 48 h before Respisure and control group. The study demonstrated the beneficial effect of coupled administration of Lydium-KLP or Methisoprinol and Respisure on the improvement of health status and the productive performance of pigs, as well as on the effectiveness of specific immunoprophylaxis of MPS in large- scale commercial breeding. The most beneficial variants of coupled administration were the simultaneous application of Lydium- KLP and Respisure, and Methisoprinol application 48 h before immunisation with Respisure.
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 107TCID50/ml. Methisoprinol (Polfa, Poland) at concentrations of 0, 100, 200, 300, 400 and 500 μg/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 μg/ml of methisoprinol in medium. The results of this in vitro study showed that methisoprinol inhibits the rhabdoviruses isolated from carp and catfish.
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