Instructions for laboratory training in General Microbiology & Immunology for students of medical faculty

МИКРОБИОЛОГИЯАЛЛЕРГОЛОГИЯ И ИММУНОЛОГИЯЛАБОРАТОРНЫЕ РАБОТЫИНОСТРАННЫЕ СТУДЕНТЫПРАКТИЧЕСКИЕ НАВЫКИПРАКТИЧЕСКИЕ ЗАНЯТИЯИНСТРУКЦИИИММУНОЛОГИЯВ издании содержатся план, программа практических занятий и основные практические навыки по общей микробиологии и иммунологии

Experimental Substantiation of Cultural Technologies Introduction into Manufacturing of Anti-Rabies Immunoglobulin

The review provides information on the major outcomes of research and development work, performed within the frames of the Federal Target Program “National system of chemical and biological safety” (2009-2014), aimed at elaboration and introduction of cultural techniques into the manufacturing of anti-rabies immunoglobulin. Described are the key phases in methodology engineering, deployed for the large-scale cultivation of fixed rabies virus, concentration of cultural liquid, quantitation of rabies virus using PCR, and immunization of producers. Obtained pilot batches of the enhanced anti-rabies immunoglobulin, complying with normative requirements to commercial formulations, testify to the effectiveness of the developed biotechnological and methodological procedures, as well as of the designed engineering-manufacturing project documentation

Optimization of Specifications for Scaled-Up Fixed Rabies Virus Cultivation (“Moscow 3253” Strain) in Vero Cell Culture

/cell, using maintenance media 199 with admixture of 0.1 % human serum albumin or 2 % bovine serum. Optimum media volume in the roll-bottle for fixed rabies virus strain, “Moscow 3253”, cultivation is 200-400 ml. It depends upon the proliferating surface area. Specifications stated above provide for the obtainment of culture liquid with rabies virus titer - 1:256 - 1:512, if assayed in ELISA. This cultural virus is recommended as a basis for immunization material obtainment with a view to produce anti-rabies immunoglobulin from equine blood serum

Divalent EuRh2Si2 as a reference for the Luttinger theorem and antiferromagnetism in trivalent heavy-fermion YbRh2Si2

Application of the Luttinger theorem to the Kondo lattice YbRh2Si2 suggests that its large 4f-derived Fermi surface (FS) in the paramagnetic (PM) regime should be similar in shape and volume to that of the divalent local-moment antiferromagnet (AFM) EuRh2Si2 in its PM regime. Here we show by angle-resolved photoemission spectroscopy that paramagnetic EuRh2Si2 has a large FS essentially similar to the one seen in YbRh2Si2 down to 1 K. In EuRh2Si2 the onset of AFM order below 24.5 K induces an extensive fragmentation of the FS due to Brillouin zone folding, intersection and resulting hybridization of the Fermi-surface sheets. Our results on EuRh2Si2 indicate that the formation of the AFM state in YbRh2Si2 is very likely also connected with similar changes in the FS, which have to be taken into account in the controversial analysis and discussion of anomalies observed at the quantum critical point in this system

Cultural Antigen in the Technology for Anti-Rabies Immunoglobulin Obtainment from Equine Blood Serum

week after immunization (specific activity is identified using neutralization reaction on the model of white mice and dot-blot immunoassay). This level of activity is sufficient for the fractioning of immune serum and extraction of anti-rabies immunoglobulin. Physicochemical and biological properties of the anti-rabies immunoglobulin, obtained with the help of cultural antigen technique, meet the requirements stated in the normative documentation on anti-rabies immunoglobulins extracted from equine blood serum. Specific activity level of experimental batches of anti-rabies immunoglobulin, obtained with the help of cultural technologies, corresponds to 242 and 214 IU/ml

Production of Rabbit Anti-Rabies Immunoglobulin Using Cultural Antigen

Justified is the possibility of application of fixed rabies virus Moscow 3253, reproduced on the cell culture Vero, as an antigen for heterologous anti-rabies immunoglobulin production. Application of the adjuvant - aluminium hydrate - is determined to be effective. Herein, by the day 73 post immunization, specific antibodies titer is ≥ 1:500 (wherein specific activity has been identified by means of neutralization test, carried out on white mice, and dot-blot immunoassay). The level of specific activity in experimental samples of anti-rabies immunoglobulin, isolated from rabbit immune serum, corresponds to 332 and 347 ME/ml. Physicochemical and biological properties of anti-rabies immunoglobulin, produced with the help of cultural antigen, fully comply with regulatory requirements specified for commercial preparation of heterologous anti-rabies immunoglobulin

ISOLATION OF GLICOPROTEID FROM THE FIXED RABIES VIRUS, STRAIN «MOSCOW 3253», AND CONSTRUCTING OF DOT-IMMUNOASSAY DIAGNOSTICUM ON ITS BASIS

Described here are the results of glicoproteid isolation from the fixed rabies virus, strain «Moscow 3253», using non-ionic detergent with subsequent chromatographic purification. The obtained antigen was demonstrated to be applicable as immunoreagent for construction of diagnosticum, by means of conjugation with colloid gold nanoparticles. The diagnosticum is meant for detection of specific antibodies in immune sera of horsesproducers, and in the preparation of anti-rabies immunoglobulin, in dot-immunoassay

OPTIMIZATION OF PRESENTATION AND CONSUMER CONTAINER OF ANTI-RABIES IMMUNOGLOBULIN OBTAINED FROM HORSE SERUM

We presented the data concerning optimization of presentation and consumer container of anti-rabies immunoglobulin obtained, from horse serum.. During the experiments, ampoules and. flasks were used for primary packaging. They were filled within 5 ml of the preparation of freeze-drying. Comparative analysis of physical, chemical and. biological properties and. molecular parameters of freeze-dried and initial forms of the immunoglobulin. was carried out. Freeze-drying was demonstrated to promote stabilization of its properties and. prevent emergence of fragments and. aggregates during anti-rabies immunoglobulin storage

Клещевой энцефалит: иммунологические показатели возможного перехода острой стадии в хроническое течение болезни

Several autoimmune diseases with chronic clinical course are characterized by detection of DNA autoantibodies in patients’ serum, while there are no such IgGs in healthy donors’ blood or in patients with acute clinical course with no evidence of chronization. Tick-borne encephalitis has not been considered this way. Several strict criteria have been applied to show that the DNase activity is an intrinsic property of IgGs from the sera of TBE patients but not from healthy donors. The relative activity of IgGs has been shown to vary extensively from patient to patient, but most of the preparations (91%) had detectable levels of the DNase activity. Polyclonal DNase IgGs were not active in the presence of EDTA or after a dialysis against EDTA, but could be activated by several externally added metal ions, with the level of activity decreasing in the order Mn2+ + Ca2+ ≥ Mn2+ + Mg2+ ≥ Mn2+ ≥ ≥ Mg2+ + Ca2+ ≥ Ca2+ ≥ Mg2+ > Ca2+, while K+ , Na+ , Ni2+, Zn2+, and Cu2+ did not stimulate DNA hydrolysis. Affinity chromatography on DNA-cellulose separated the DNase IgGs into many subfractions with various affinities for DNA and very different levels of the relative activity. Possible reasons for catalytic diversity of polyclonal human Abzs are discussed.Ряд аутоиммунных заболеваний с хроническим течением характеризуются обнаружением в крови больных ДНКаутоантител, в то время как их не содержит кровь здоровых доноров или пациентов с острым течением заболеваний, незначительным нарушением иммунного статуса, без определенной склонности к переходу в хронический процесс. Клещевой энцефалит (КЭ) не рассматривался с этих позиций. Предварительно для данной работы проведен поиск достаточно точных критериев обнаружения ДНК-активности антител иммуноглобулина (Ig) G из сыворотки крови больных КЭ и здоровых доноров. Показано, что относительная активность антител IgG значительно варьирует у пациентов, но большинство образцов (91%) имели определяемый уровень ДНКазной активности. Поликлональные ДНКазные антитела IgG не активировались в присутствии ЭДТА или после диализа с ЭДТА, но могли активироваться некоторыми добавленными ионами металлов с уровнем активности, уменьшающимся в ряду Mn2+ + Ca2+ ≥ Mn2+ + Mg2+ ≥ Mn2+ ≥ Mg2+ + Ca2+ ≥ Co2+ ≥ Mg2+ > Ca2+, в то время как K+ , Na+ , Ni2+, Zn2+ и Cu2+ не стимулировали гидролиз ДНК. Аффинная хроматография на ДНК-целлюлозе разделила ДНКазные антитела IgG на множество субфракций с различным сродством к ДНК и очень разными уровнями относительной активности. Возможные причины каталитического разнообразия поликлональных человеческих аутоантител обсуждаются

A comparison of the radiosensitisation ability of 22 different element metal oxide nanoparticles using clinical megavoltage X-rays

Background: A wide range of nanoparticles (NPs), composed of different elements and their compounds, are being developed by several groups as possible radiosensitisers, with some already in clinical trials. However, no systematic experimental survey of the clinical X-ray radiosensitising potential of different element nanoparticles has been made. Here, we directly compare the irradiation-induced (10 Gy of 6-MV X-ray photon) production of hydroxyl radicals, superoxide anion radicals and singlet oxygen in aqueous solutions of the following metal oxide nanoparticles: Al2O3, SiO2, Sc2O3, TiO2, V2O5, Cr2O3, MnO2, Fe3O4, CoO, NiO, CuO, ZnO, ZrO2, MoO3, Nd2O3, Sm2O3, Eu2O3, Gd2O3, Tb4O7, Dy2O3, Er2O3 and HfO2. We also examine DNA damage due to these NPs in unirradiated and irradiated conditions. Results: Without any X-rays, several NPs produced more radicals than water alone. Thus, V2O5 NPs produced around 5-times more hydroxyl radicals and superoxide radicals. MnO2 NPs produced around 10-times more superoxide anions and Tb4O7 produced around 3-times more singlet oxygen. Lanthanides produce fewer hydroxyl radicals than water. Following irradiation, V2O5 NPs produced nearly 10-times more hydroxyl radicals than water. Changes in radical concentrations were determined by subtracting unirradiated values from irradiated values. These were then compared with irradiation-induced changes in water only. Irradiation-specific increases in hydroxyl radical were seen with most NPs, but these were only significantly above the values of water for V2O5, while the Lanthanides showed irradiation-specific decreases in hydroxyl radical, compared to water. Only TiO2 showed a trend of irradiation-specific increase in superoxides, while V2O5, MnO2, CoO, CuO, MoO3 and Tb4O7 all demonstrated significant irradiation-specific decreases in superoxide, compared to water. No irradiation-specific increases in singlet oxygen were seen, but V2O5, NiO, CuO, MoO3 and the lanthanides demonstrated irradiation-specific decreases in singlet oxygen, compared to water. MoO3 and CuO produced DNA damage in the absence of radiation, while the highest irradiation-specific DNA damage was observed with CuO. In contrast, MnO2, Fe3O4 and CoO were slightly protective against irradiation-induced DNA damage. Conclusions: Beyond identifying promising metal oxide NP radiosensitisers and radioprotectors, our broad comparisons reveal unexpected differences that suggest the surface chemistry of NP radiosensitisers is an important criterion for their success