Aim. To investigate influence of the Deschampsia caespitosa L. and Calamagrostis epogeios L. herbal extract (HE) and its main components – quercetin derivatives – on the DNA and RNA synthesis; to per- form comparative analysis of their antiviral activities. Methods. The model transcription system of bacteriophage T7 (RNAP T7) and polymerase chain reaction (PCR) were used as test systems. For antiviral studies there were employed models of influenzal and herpetic infections and hepatitis C. Results. It was shown, that HE, its major component 7, 3'-dimetoxyquercetin (DMQ) its minor component 5,7,3',4'-tetrametho- xyqeercetin (TMQ) effectively inhibited the RNA synthesis in the RNAP T7 system with the corresponding values of IC50 – 0.07, 4 and 1 µg/ml. Minimal concentrations of the agents IC90, completely inhibiting PCR, were – 8, 30 and 40 µg/ml for HE, TMQ and DMQ, accordingly. All the studied preparations revealed high multiple antiviral activities against the RNA- and DNA-containing viruses. Conclusions. Antiviral activity of the herbal extract is likely determined by the combined action of its components, TMQ and DMQ including. Bearing in mind multi-target profile of quercetin derivatives, an assumption may be made, that antiviral mechanism of the agents studied consists in blocking of enzymatic systems of the RNA and DNA synthesis
Background: Herpes simplex virus (HSV) is prevalent in today’s world population, and there is evidence of potential HSV reactivation in patients with immune deficiency induced by acute stroke. However, the data on the use of antivirals in the setting of stroke are scarce. The aim of this study was to evaluate the reactivation of HSV-1 in patients with stroke, using several methods, and to assess the efficacy of acyclovir in the treatment of experimental stroke. In the employed methodology, PCR and dot-ELISA were used to detect the occurrence of HSV-1 in patients with acute stroke. White mice were infected with HSV-1 and experimental stroke was simulated. The infected mice with stroke were subdivided into two groups: one of them received no treatment, while the other one was treated with acyclovir. The level of HSV-1 reactivation was determined by the methods used in human patients. The brain tissue of experimental animals was also subjected to morphological and morphometrical study. The results of such work reveal that, by the applied serological method, HSV-1 was found in all patients with stroke. Herein, the increased level of HSV-1 was seen in the brain tissue and blood in 100% of the experimental infected animals. However, the use of acyclovir suppressed reproduction of HSV-1. Hence, it can be concluded that clinical and laboratory studies have demonstrated the different sensitivity of Dot-Elisa and PCR, with the former being more sensitive. Moreover, the use of acyclovir in the experiment inhibited viral reproduction and further development of viral infection. Still, chemic lesions in the brain persisted.
A wide range of interleukin-7 (IL-7) biological effects suggests that application of appropriate preparations in clinical practice will stimulate immunity in patients with lymphocytic exhaustion or autoimmune diseases. Studies are being conducted for Il-7 based preparations aimed at restoration of the immune system of patients with immunodeficiency of different origins. Pseudomonas aeruginosa is an important pathogen, which causes nosocomial infections in hospitalized patients. Infection factors, affecting the immune status of the host, play a key role. A promising and relevant scientific endeavor is the study of the effect of recombinant as an adjunct therapy in wound infections caused by P. aeruginosa. the aim of this study was to evaluate the effectiveness of rIL-7 use in P. aeruginosa wound infection in mice. The experiments were conducted using a standardized rIl-7 preparation, P. aeruginosa strain and 20 white non-inbred mice. the preparation of rIL-7 after all stages of purification was characterized by the content of ballast proteins and impurities via electrophoresis in a polyacrylamide gel in reducing conditions, and its biological activity was evaluated in the Mtt test by means of proliferation of peripheral blood mononuclear cells. In the mice, the fur was removed, the neck nape was intentionally injured and P. aeruginosa bacteria were injected into the wound of each animal (0.1 ml of suspension with a bacterial cell concentration of 0.08×10 9 cells/ml). Starting from the 2 nd day, bacterial examination of the wound material was carried out daily. Starting from the 3 rd day, the mice (experimental group, n = 10) were intraperitoneally administered 5 μg (0.1 ml) of the rIL-7 preparation. In the control group of animals (n = 10), the rIl-7 preparation was not administered. In 80% of experimental animals (administered the rIl-7 preparation), the healing of wounds and elimination of the pathogen of purulent inflammatory infection P. aeruginosa occurred on the 7 th day. on the 9th day from the beginning of wound infection, wound healing and elimination of P. aeruginosa occurred in all experimental mice. In 60% of mice from the control group (did not receive treatment with rIl-7), wound healing and the elimination of P. aeruginosa occurred on the 9th day. Wound healing and elimination of P. aeruginosa in all mice of the control group occurred on the 14th day. thus, in mice treated with rIl-7, wound healing and elimination of the pathogen occurred 5 days earlier than in mice from the control group . Subsequent studies may be aimed at developing protocols for the treatment of wound infections using an rIl-7 preparation in patients with a compromised immune system therefore, rIl-7 is a promising preparation for the treatment of complex wound infections. k e y w o r d s: recombinant human interleukin-7, Pseudomonas aeruginosa, wound infection, therapy.
Immunoglobulins M (IgMs) are the evolutionally oldest class of antibodies in higher eukaryotes. This pool of antibodies is one of the first to appear in humans and begins to be-synthesized at the early stages of the neonatal period. Most of the repertoire of IgMs of the newborns consists of the so-called «natural», or «naive», antibodies synthesized by the body without external antigenic stimulation. In addition to the classical functions of human immunoglobulins M (such as antigen recognition and initiation of innate immune responses), antibodies of this class exhibit a variety of non-canonical functions. The non-canonical functions are the action of antibodies as agonists/antagonists of various receptors, cleavage of antigen due to the catalytic activity of IgM, direct inactivation of pathogens in the absence of effector cells and molecules, etc. The aim of this work was to study and compare the antiviral activity of total preparations of immunoglobulin M of newborns and adults, obtained from umbilical cord and venous blood sera, on the LLC-PK1 cell line model infected with the transmissible gastroenteritis virus (TGEV). In addition, in the course of the studies, a decision was made to investigate the effect of combined preparations of immunoglobulin M on the change in signal transduction in the epidermal growth factor receptor as one of the mechanisms of TGEV internalization during infection of target cells. Methods. Highly purified preparations of total IgM of adults or newborns were obtained using the methodologies of sequential salt fractionation and affinity chromatography. This work used the model of the interaction of the transmissible gastroenteritis virus with cells of the LLC-PK1 line and the monitoring of changes in the phosphorylation state of the epidermal growth factor receptor of these cells during virus infection to study the effect of human IgM on the internalization of the virus and its interaction with the receptor system of the host cell. The degree of cytopathogenic effect of the virus was determined visually by changes in cell morphology. The mean infectious dose for transmissible gastroenteritis virus in the cell culture of the LLC-PK1 line was determined by the Reed-Muench method. Analysis of changes in the phosphorylation of the epidermal growth factor receptor was performed using the Western-blot analysis method. Results. The addition of a total high-purified sample of human IgM reduces the degree of efficiency of TGEV infection of the LLC-PK1 cell line and modulates the phosphorylation levels of these cells. Conclusions. The total preparations of IgM obtained from human venous blood of adults and from umbilical cord blood of newborns can affect the internalization of the transmissible gastroenteritis virus in the LLC-PK1 cell line. The original model of virus (TGEV) — cell line (LLC-PK1) was applied and tested to study the effect of native total preparations of immunoglobulin M on the internalization of the virus into the cell. The obtained data can be useful in further studies for a better understanding of the process of development and functioning of the immune system of newborns.
The aim: To study the effect of a high-fat diet (HFD) on the structural changes in the aortic intima in intact and HSV-1-infected mice using Scanning electron microscopy (SEM) and Transmission electron microscopy (TEM). Materials and methods: In experiments Balb/c mice were infected with the HVS-1 and fed high-fat diet and 12 weeks later aortic ultrastructure was examined by SEM and TEM methods. The animals were subdivided into four experimental groups: 1st group – HSV-1-infected animals; 2nd – animals consuming high-fat diet (HFD); 3rd – infected animals that were subsequently consuming a high-fat diet (HSV / HFD); 4th – animals consuming a high-fat diet that were subsequently infected with HSV-1 (HFD / HSV) (n = 6); and control group – intact animals. Results: HVS-1 impaired ultrastructural changes in aorta greater than high-fat diet and HVS-1 alone (higher density of lipid inclusions in the subendothelial space, necrosis of endothelial cells), and infection of mice after high-fat diet ended 100% mortality. The formation of atheroma in the aortic wall during HFD was not detected, but the initiative manifestations of atherogenesis have been identified and restricted in the aortic intima. These structural changes included lipid inclusions in the subendothelial space, cell damage and destruction, which lead to an increase cellular detritus in the 3rd (HSV / HFD) group. Conclusions: HSV infection potentiates the accumulation of lipid inclusions in the aortic intima during a HFD, facilitates infection and contributes to the development of acute infection.
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