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The ability of macrophages to phagocyte granulocyte debris ("cleaning" reaction) is evolutionary developed and represents the mechanism of cellular resistance preceding immunity. Activation of the mononuclear phagocyte system and acquisition by the macrophages of the capacity to the intracellular chlamydia destruction as well as other infectious agents parasitizing in the intact cells occurs in the process of the granulocyte debris phagocytosis. This phenomenon termed by the author as a resorptive cellular resistance is one of host barrier mechanisms.

Transitory age-associated deficiency of the granulocyte cationic proteins as a cause of a phagocytosis disturbance, loss of the phagocyte cell ability to kill phagocytosed bacteria is established. Quantitative indexes of myeloperoxidase content, elastase and catepsin G in the neutrophil granulocytes of newborns and laboratory animals are determined. Hypothesis is developed on the transitory age-associated immunodeficiencies of the neutrophil granulocyte system as one of the endogenic risk factors in the development of infectious diseases during early postnatal period.

The knowledge of the molecular level of the general pathologic processes is a prerequisite for new concepts realized in the practice. The molecular destructive processes, programmed cell death and inflammation are considered as examples. The hypothesis of the catastrophe of errors is reviewed, the results of studies on the limited proteolysis reaction as well as intracellular destructive processes compatible with the life of cell are summarized. The concept of nonphagocytic tissue resistance and resorptive cellular resistance as a peculiar type of nonspecific protective reactions in the infectious inflammation is substantiated.

The data of the author's own studies and those from the literature concerning morphological manifestations of antimicrobial activity of cation proteins of polymorphonuclear leukocytes (PNL) in phagocytosis and inflammation are summarized. A previously unknown type of secretion of PNL lysosomal proteins termed proexocytosis or pseudo-degranulation is characterized. A hypothesis on formation of nonphagocytic tissue resistance in inflammation foci is proposed. A lysosomal cation test expanding the possibilities for evaluation of the efficacy of treatment and prognosis of the course of diseases of the broncho-pulmonary system, purulent surgical diseases, infectious-inflammatory diseases of the urogenital organs, burn disease, parodontosis is proposed for the clinical practice. Ways for the development of methods for early diagnosis of hereditary lysosomal microdefects of PNL are outlined. Morphological manifestations of intracellular parasitising of microbes in macrophages are analysed. The problem of unity and interaction of granulocytic, macrophage, and lymphoid systems in hypersensitivity and inflammation is discussed.

The functional activity displayed by polymorphonuclear leukocytes (PL) of the peripheral blood and gastric mucosa, including ulcer margins, was studied in 50 rabbits with ulcer experimentally induced by the Okabe method and in 25 intact animals. The peripheral blood PLs were found to show substantially increased lysosomal cation proteins (LCP), enhanced alkaline phosphatase and decreased peroxidase activity. The changes in the gastric mucosa, particularly in its damaged area, were characterized by a marked microcirculatory bed response and formation of lymphocytic and, later on, pronounced neutrophilic infiltrates. The tissue PL therewith contained abundance of glycogen and LCP and intensively secreted granules. The maximum of these manifestations as viewed from PL coincided with the ulcer formation period (on days 2-3 of the experiment). When the ulcer had developed, the levels of LCP and glycogen in PL became lower. It was concluded that PL made an important contribution to ulcerogenesis.

Interactions of the causative agents of meningopneumonia and ovine abortion with pulmonary macrophages in experimental infection in white mice were studied by histological, immunofluorescent and electron microscopic methods. The causative agent of meningopneumonia did not multiply in colonies and caused toxic lesions in the cells and formed cytoplasmic oxiphilic inclusion bodies. Being the product of cell degeneration, these oxiphilic inclusion bodies were conducive to long-term survival of the cell and slow release from them of elementary bodies of meningopneumonia agent. The agent of ovine abortion multiplied in colonies without toxic affection of the cells or formation of oxiphilic inclusion bodies. It is suggested that oxiphilic inclusion bodies represent morphological manifestation of the intracellular toxic effect of rickettsial and ornithosis-like agents possessing dissociated (non-colonial) mode of multiplication.

Neutropholic leukocytes of aseptic peritoneal exudate of rabbits differed from neutrophilic leukocytes of the blood in greater variability in the content of cationic proteins. Some lysosomes ceased to be stained on cationic proteins, which produced a false impression of degranulation of leukocytes. Ultramicroscopically, there was observed a "clearing" of the lysosomes content and its outflow into cytoplasm. Lysosomes acquired the shape of spherical "holed" bodies and were transformed into clear vacuoles with remnants of the electron-dense material. The rupture of lysosome membranes led to an "explosive" type of liberation of the lysosome content into the cytoplasm. Partly degranulated leukocytes remained vital and activity moved about in a homologous serum. Following contacts with histone F1 leukocytes lost cationic proteins. The liberation of cationic proteins from leukocytic took place without destruction of leukocytes and lysosomes themselves.

A study was made of the influence of UV-irradiation (254 nm) of blood in vitro, of the autotransfusion of UV-irradiated blood (AUVIB), and of the mixture of UV-irradiated and intact blood in vitro on the content of bactericidal cation proteins (CP) in blood neutrophil of calves suffered from dyspepsia and broncho-pneumonia. Age differences were noticed in CP contents and their decrease in neutrophils following AUVIB in vivo and administration of the mixture of blood in vitro. The decrease in cell CP contents is presumably due to neutrophil degranulation and CP release into the blood plasma. Since the initial mechanisms of neutrophil degranulation are located on the cell surface, the CP release is supposed to result from a membranotropic effect of UV-irradiated blood on the intact autologous blood. This effect may explain the increase in nonspecific resistance of organism after the AUVIB, being one of the main therapeutic phenomena of the AUVIB-therapy.

The authors consider that quantitative and qualitative characteristics of phagocytosis can be given with the help of lysosomal-cationic test (LCT) at different periods of acute purulent lactation mastitis. LCT gives prognosis of the course of the disease and a reliable estimation of the treatment efficiency.

It was found in studying the antimeningococcus activity of the leukocytic and thymus histones and their fractions that both histones were capable of neutralizing in vitro the activity of the causative agent of meningopneumonia (MP). The neutralization effect was chiefly associated with the F3 fraction rich in arginine and depended on the duration of the histone fraction contact with the MP causative agent, the weight concentration of the histone and the pH of the incubation medium.

A cytochemical lysosome-cation test is proposed as suitable for use in diagnostic laboratories. The test is based on the detection of cationic protein and determinations of the fate of phagocytized and extracellular bacteria in inflammation foci. A method for simultaneous cytochemical detection of myeloperoxidase and cationic proteins in neutrophilic and eosinophilic granulocytes was developed. Using this method, a previously unknown variety of lysosomal granules devoid of peroxidase was detected in neutrophilic promyelocytes. The lysosomal cation test was shown to be useful for improved morphological diagnosis of leukemias, for the determination of the fate of phagocytized and non-phagocytized bacteria in inflammation foci, and of the level of nonspecific and infection resistance of the host.

A method of histochemical identification of cationic proteins, peroxidase, viable and killed bacteria in semithin sections of biopsy samples is suggested. The method is based on dying of araldite-embedded semithin sections with a buffered alcohol solution of fast green (pH 8.15) and on peroxidase identification with O-dianisidine. The method suggested can be used for distinguishing occult forms of infectious gastritis, evaluation of secretory and barrier function of gastric epithelium, selective dying of leucocytes in tissues, revealing of viable and non-viable bacteria in the infective foci.

A method is suggested for a histochemical and immunofluorescent detection of lysosome cation proteins in the histologic paraffin sections of biopsies. The method is based on the staining of histologic sections obtained from the material fixed in glutaraldehyde buffered with an alcohol solution of a fast green (pH 8.1-8.2) and on the detection myeloperoxidase and lactoferrine by means immune serum by Coons's method. 0.25% solution of azur A (B, C) is used for staining the cell nuclei and viable bacteria.

Phagocytosis begins with exocytosis--"extrarapid" discharge of bactericidal proteins and factors of permeability into the extracellular medium. A viewpoint was put forward on an "avalanch-like" character of the outcome of cationic proteins from leukocyte granules in inflammation and their participation in formation of a nonphagocytic type of resistance. In phagocytosis bacteria perish due to the myeloperoxidase system, lysozyme, lactoferin and nonenzymic cationic proteins. Hereditary deficit of the above-mentioned substances leads to intraleukocytic microbicidal insufficiency, a drastic decrease in the nonspecific resistance of the organism and to development of fatal granulomatous disease, and to other forms of pathology associated with genetic defects of the bactericidal systems of leukocytes.

The article deals with data on lysosomal cationic proteins. Many properties of cationic proteins of leucocytes justify the assumption about their possible genetic affinity to nuclear histones. A high antibacterial activity of cationic proteins justified a more profound investigation into their role in phagocytosis and in formation of anti-infectious resistance in inflammation foci. The question concerning cationic proteins as mediators of inflammation is discussed. The suggested methods for staining cationic proteins may be used in determination of the fate of phagocytized bacteria, in studies of cytochemistry of cationic proteins of leucocytes, in identifying viable and perished bacteria in inflammation foci in bacterial infectious processes.

Lysosomal secretion of neutrophilic granulocytes has been studied in normal subjects (healthy males and females aged 20-50), pregnant women and burnt patients. A significant rise in lysosomal secretory activity of neutrophilic granulocytes was recorded in the 3rd trimester of pregnancy. Low lysosomal secretion can serve an indicator of poor prognosis in patients with a severe burn trauma.

Clinico-morphological tests for the estimation of neutrophil granulocyte function were compared by their reproducibility, reliability and availability for standard clinical laboratories, and the results were analysed. Lysosomal-cationic test was offered for clinical practice, as it gives qualitative and quantitative assessment of the phagocytosis process, allows prediction of complications before their clinical onset, performance of rapid evaluation of the therapy used, reveals shifts in the level of non-specific resistance of the body. Methods of application of lysosomal-cationic test in the pathoanatomic practice have been developed for the study of intravital biopsy samples and surgical material.

A study was made of the influence of cation proteins from the lysosomes and nuclei of rabbit polymorphonuclear leukocytes on the survival of the causative agent of meningopneumonia in vitro. As revealed, the greatest antibiotic activity was possessed by the nuclear histones and the low molecular basic proteins of lysosomes. Biological significance of the detected effects is discussed.

Lactoferrin, a non-heme iron-binding protein was isolated from pig neutrophils. The purification procedure included initial extraction of the protein in the presence of cetyltrimethylammonium bromide followed by chromatography on carboxymethyl-cellulose and Sephadex G-100. The thus obtained protein was found to be homogeneous on polyacrylamide gel (PAAG) electrophoresis at acidic values of pH. PAAG electrophoresis in the presence of sodium dodecyl sulfate revealed a single component with a molecular weight of 75 000-80 000. The resulting protein is capable of binding two atoms of iron molecule. The absorbance spectra for the pig neutrophil lactoferrin are identical to those for cow milk lactoferrin in the visible region and have a maximum at 465 nm. The amino acid composition of pig lactoferrin was determined. Isoelectric focusing of the protein obtained in a PAAG stabilized pH gradient revealed a component with pI of about 6.8. A single precipitin line was observed with rabbit antipig lactoferrin when examined by immunodiffusion. No immunological cross-reactions were observed between pig lactoferrin and bovine lactoferrin.

The dymanics of formation of macrophagal granulomas in the liver of albino mice was studied after intravenous infection with the causative agent of ornithosis. In formation of granulomas macrophages underwent the stage of reutilization of the leukocyte disintergration and became resistant to the causative agent of ornithosis. Together with the leukocyte remants histones and cation proteins could penetrate into the macrophages cytoplasm; these displayed a high antironithosis activity in vitro. An increase of macrophage resistance to the causative agent after reutilization of the products of leukocyte disintegration apparently sreved as one of the mechanisms of resistance formation in the foci of inflammation in ornithosis.

Clinico-morphological test for the quantitative evaluation of exudative and cell response in the diagnostical inflammatory focus in various forms of human pathology is developed. The values in practically healthy persons are established. The possibilities of the test use for the prognosis of disease and the development of postoperative complications in patients with cold trauma are shown.

A method has been proposed for detection of lysosomal cation proteins of polymorphonuclear leukocytes in histological paraffin sections. The method is based on staining of histological sections of Brank fluid-fixed material with an alcohol solution of green fast at pH 8.1. A 0.25% aqueous solution of azure A was used for poststaining of cell nuclei and viable bacteria.

Some methodological drawbacks of the previously offered by the author method for speedy staining of lysosomal cation proteins have been eliminated. The method is shown to be applicable for histochemical staining of lysosome cation proteins of human polymorphonuclear leucocytes. Staining is carried out on blood smears and preparations-impressions from the surface of incision on the organ under study. The improved method for staining of cation proteins can be used in determining the fate of cytophagous bacteria, in cytochemistry of cation proteins of polymorphonuclear leucocytes, in detecting vital and perished bacteria in inflammation foci in bacterial infectious processes.

Repeated intranasal injections of increasing doses of bacterial lipopolysaccharide pyrogenal resulted in accumulation of numerous macrophages in the upper and middle parts of the mouse lungs; these macrophages were filled with leukocyte lysosomes. In these animals dilysosomal macrophages increased local resistance to the agent of enzootic ewe abortion and thus could change the lethal form of induced pneumonia into the nonlethal one.

Some physico-chemical properties of human and pig lactoferrins from milk and neutrophilic granulocytes were compared. It was shown that the lactoferrins from different cell and tissue sources of the same species (humans or pigs) are identical in terms of electrophoretic mobility, molecular weight, iron-binding capacity, absorbance spectra, amino acid and sugar compositions and peptide maps. Human and pig lactoferrins show a high degree of structural homology (approximately 50%), but are immunochemically different.

Two cation proteins free of enzymatic activity, containing one and four peptides (CP-1 and CP-4, respectively), were studied. The cation protein consisting of four peptides proved to be the most effective inhibitor of the nonenzymatic fibrinolytic activity of heparin-containing complexes. Both these cation proteins inhibited similarly the anticoagulation activity of blood plasma. The drugs, like cation proteins, appear to be potentially important for clinical practice under conditions of excessive hemorrhage caused by elevated level in circulation of complexes containing heparin and proteins.

A model system has been developed (B. megaterium + granulocyte) imitating the mechanisms of phagocytosis of filamentous forms of bacteria by neutrophilic granulocytes. In the capture of multicellular microbial filaments by one or several granulocytes predominate the mechanisms of cell membrane invagination and formation of exocytic secretory system providing the inhibition of the activity of the phagocytized agent and release of microbicidal factors and enzymes into the extracellular environment. The importance and frequency of occurrence of the observed phenomenon in different pathological forms have not been elucidated.