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Purification and characterization of an anti-inflammatory factor (AIF) from bovine serum.

An anti-inflammatory factor (AIF) was highly purified from normal bovine serum. The purified AIF was a polysaccharide which was formed from a low molecular substance Pro-AIF by macromolecularization. AIF showed potent inhibitory activities against carrageenin induced edema in rats and PMN-leucocyte chemotaxis. It showed no inhibition in cutaneous reaction with serotonin, bradykinin or a mediator releaser compound 48/80. No inhibition was also observed in thermally induced pain, reversed passive Arthus reaction and adjuvant polyarthritis in rats. With these results, the role of AIF in inflammation was discussed.

Analgesics

Does the pro-inflammatory factor in lymphocytes (LpIF) explain the role of these cells in acute inflammation?

Lymphocytes produce a pro-inflammatory factor, which modulates the development of acute inflammation. Injection of lymphocytes or products, obtained from either rats, dogs or rabbits, caused a restoration of inflammatory responses in leukopenic rats which are hyporeactive to various inflammatory stimuli. In vitro incubation of viable lymphocytes with homologous and heterologous anti-lymphocyte sera abolished the ability of the cells to restore the inhibited inflammatory reactions.

Animals

The Anti-Osteoporosis Effects of Panax japonicus via Downregulation of Inflammatory Factors: A Network Pharmacology and Ovariectomized Rat Model Study.

OBJECTIVE: Osteoporosis is a major and growing public health problem characterized by decreased bone mineral density and destroyed bone microarchitecture. Panax japonicus has been clinically used in the treatment of bone diseases, especially osteoporosis. However, there is a lack of study on the mechanism of osteoporosis treatment with Panax japonicus. MATERIALS AND METHODS: A network pharmacology approach was employed to identify the targets of osteoporosis and Panax japonicus. Cytoscape 3.7.2 and DAVID were used to visualize the pharmacological mechanism of Panax japonicus in treating osteoporosis by building up compound-target and protein-protein interaction (PPI) networks and conducting Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. An ovariectomized SD rat osteoporosis model was used to assess the potential therapeutic effect of Panax japonicus in vivo. The biomechanical properties, pathological changes, inflammatory cytokines, bone density, and bone microstructural parameters in rat bone tissue were carefully measured. The biochemical markers of bone metabolism in serum were detected by Enzyme-Linked Immunosorbent Assay (ELISA). RESULTS AND DISCUSSION: Fifty-two active components and sixty-five target genes of Panax japonicus involved in the treatment of osteoporosis were identified. The PPI network revealed IL-6, TNF, NR3C1, IL-1β, CASP3, ESR1, PGR, and AR to be involved in the treatment of osteoporosis with Panax japonicus. Chikusetsusaponin IVa and Radix ginsenoside-Ro were the main saponins found in Panax japonicus. Panax japonicus was found to exert potent preventive effects on osteoporosis by maintaining biomechanical properties, increasing bone mineral density, and protecting the trabecular microstructure in an ovariectomized rat osteoporosis model. Panax japonicus hindered the initiation of osteoporosis induced by ovariectomy by regulating bone metabolism and downregulating the expression of IL-6 and TNF-α. CONCLUSION: Panax japonicus was found to contain 52 compounds and 65 targets in the treatment of osteoporosis. The administration of Panax japonicus could mitigate osteoporosis in rats induced by ovariectomy, and one of the mechanisms was associated with downregulating the expression of inflammatory factors.

Animals

An allograft inflammatory factor enhances sperm viability by modulating intracellular calcium in oyster Crassostrea gigas.

As an important aquaculture bivalve, the Pacific oyster Crassostrea gigas faces severe constraints in artificial reproduction, where low sperm motility often leads to fertilization failure and limits the sustainable development of the oyster aquaculture industry. In the present study, the variation of sperm from different oyster individuals was observed, and high-quality sperm possessed intact, elongated flagella with no structural abnormalities, while low-quality sperm showed shortened flagella with frequent tangling or coiling defects. Transcriptomic analysis comparing high- and low-quality sperm revealed significantly reduced expression of genes associated with sperm motility and release (CgAIF1, CgAchR, CgSEX), sperm quality and development (CgEP4, CgIFi2b), and cryoprotection (CgISPs) in low-quality sperm. Notably, an allograft inflammatory factor (designed as CgAIF1) encoding EF-hand domain, known as Ca2+ binding activity, was among the most significantly downregulated in low-motility sperm. CgAIF1 is highly expressed in haemocytes, ganglia, and gonads of oysters. Incubation with the recombinant AIF1 protein (rCgAIF1) significantly improved sperm curvilinear velocity, thereby enhancing the overall motility of C. gigas sperm. Furthermore, rCgAIF1 incubation increased intracellular Ca2+ levels (2.13-fold at 30 min, 2.71-fold at 60 min) and superoxide dismutase (SOD) activity (1.44-fold at 30 min, 1.24-fold at 60 min) in sperm, suggesting potential roles in calcium homeostasis regulation and antioxidant defense. In conclusion, this study demonstrates that CgAIF1 significantly enhances motility of oyster sperm, providing a scientific basis for artificial breeding and seed production in oyster aquaculture.

Animals

The role of bacterial contamination in the isolation of apparent anti-inflammatory factors from rabbit anti-lymphocytic serum.

1 Rabbit anti-guinea-pig lymphocytic serum was fractionated by gel filtration to obtain partially purified materials possessing anti-inflammatory activity. The pharmacological properties of these materials were then studied. 2 Two fractions were found which reproducibly contained significant activity. One of these activities caused inflammation at the site of injection and was associated with high molecular weight protein (2008000). The other activity was found in low molecular weight fraction but was shown to be due to small amounts of endotoxin from Gram negative bacteria. These organisms contaminated the fractions in spite of the recommended precautions for gel filtration having been taken. 3 The endotoxin-containing fraction completely abolished leucocyte infiltration into the rat foot which had been injected with kaolin. It had no apparent effect on circulating haemolytic complement. It caused maximal elevation of serum 11-hydroxycorticosteroid concentrations and was found to cause the release of pharmacologically active amines. Many of the previously reported naturally occurring anti-inflammatory substances have similar pharmacological properties to those of the endotoxin-containing fraction. 4 It was concluded that doubt will exist about the presence of anti-inflammatory factors in mammalian body fluids unless stringent precautions are taken to exclude measurable bacterial contamination. 5 These experiments also cast doubt on the validity of accepted procedures for excluding microbial growth from columns used in the fractionation of serum.

11-Hydroxycorticosteroids

Inflammatory factors produced by sensitized guinea-pig peripheral blood lymphocytes.

The supernatants obtained from stimulated tuberculin-sensitive guinea-pig peripheral blood lymphocytes contain factors that induce a cutaneous inflammatory response in normal guinea-pigs similar to the tuberculin reaction and inhibit the migration of normal guinea-pigs peritoneal exudate cells. There appears to be a correlation between the presence of in vitro migration inhibitory activity and inflammatory activity in vivo.

Animals

Naturally occurring anti-inflammatory factors in the synovial fluids of patients with rheumatic disease and their possible mode of action.

Evidence has been shown that pathological synovial fluid contained a substance capable of stabilizing rat liver lysosomes which was partly inactivated by treatment with trypsin and by storage. Such synovial fluid also appeared to contain a substance which labilized lysosomes and which was more stable than the stabilizing substance. (2) The lysosomal stabilizing substance described above was nondialysable and migrated electrophoretically with the alpha and beta globulins to which class it has been tentatively ascribed. (3) Pathological synovial fluid contained proteases which were active at acid pH and at neutral pH. It also appeared to contain a substance capable of inhibiting these proteases. (4) Alpha2-Macroglobulin has been detected in pathogenic synovial fluid.

Alpha-Globulins

Inflammatory reactions and chemotactic factors.

Inflammatory reactions are mediated by a large number of chemical substances called chemical mediators, including chemotactic factors, which are released by immunologic as well as non-immunologic mechanisms. Chemotactic factors which act indirectly on target tissue can not only enhance inflammatory reactions (NCF-C) but may also eliminate the reactions (ECF-A).

Anaphylatoxins

[Origin of a mitogrenic factor for cultivated macrophages (IMF: Inflammatory Mitogenic Factor) found in exudates of non-specific acute inflammation].

The mitogenic activity of inflammatory exudate obtained from irradiated Rats is reduced. After transfer of bone marrow syngeneic cells into irradiated Rats this mitogenic activity is further decreased, while after transfer of thymic cells it is increased. It is postulated that the mitogenic activity of inflammatory exudate could be related to thymic cells and that T lymphocytes may be involved in non specific-inflammatory reactions.

Acute Disease

Effect of soluble plaque factors on inflammatory reactions in the dental pulp.

The present investigation was undertaken to determine whether soluble factors from human dental plaque are capable of inducing or mediating pathological alterations in the dental pulp. The experiment was performed in three monkeys and involved 40 teeth in all: 20 test and 20 control teeth. Class V cavities were prepared on the buccal surface close to the marginal gingiva. In the test cavities an extract of human dental plaque was applied once very 5 min during an 8-h period. The cavities of the contralateral teeth were treated in an identical manner but with Ringer's solution. The animals were sacrificed 10 or 32 h after the initiation of the test. Two hours prior to sacrifice the animals were injected with colloidal carbon and then perfused with 10% neutral buffered formalin. Histologic examination demonstrates that the pulp tissue response to topical application of a plaque extract was characterized by (1) increased degree of carbon retention - indicative of vascular exudation and (2) enhanced rate of emigration of neutrophils and monocytes.

Animals

Emotional factors in inflammatory bowel disease.

Inflammatory bowel diseases are not caused by emotional conflicts, but emotional factors influence both the pathogenesis and the course of the illness. Establishing a trusting physician-patient relationship is important in helping the patient to (1) avoid denial of illness or chronic illness behavior, (2) understand symptoms and diagnostic and treatment procedures, and (3) cope with loss of a key person or the threat of lessened self-control.

Adolescent

Anaphylactoid-inflammation-promoting factor. An insulin-induced factor derived from non-sensitized lymphocytes increases anaphylactoid inflammation in rats.

The experiments reported here indicate that, when exposed to insulin, viable lymphocytes rapidly released into the incubation medium a factor capable of increasing the dextran-induced anaphylactoid reaction, but having no effect on the inflammatory response evoked by 5-HT. This pro-inflammatory factor was shown to be elaborated by cell suspensions derived from lymph nodes of rats, rabbits, pigs or calves as well as from human tonsils. Thymus cells showed no such activity. The pro-inflammatory factor was termed as anaphylactoid-inflammation-promoting factor (AIPF). Its production depended upon the dose of insulin, and the time of exposure. AIPF was found to have an elution pattern in Sephadex G-100 gels similar to that of BSA (67,000 daltons). The activity was abolished by heat or incubation with DNase or a-chymotrypsin, but was not influenced by RNase. AIPF by itself did not induce increased vascular permeability, and proved to be distinct from the permeability factors present in the lysate of lymph node cells.

Adrenalectomy

The course of carrageenin-induced iridocyclitis in rabbits.

On the basis of biochemical indices in the aqueous humor of the eye strictly related to intensity of inflammation, and morphologic observations with the slit-lamp microscope, dynamics of experimental carrageenin-induced inflammation of the anterior chamber of the eye was studied. Carrageenin-induced inflammation was characterized by intensity of changes in the aqueous humor within a few hours after infection of the inflammatory factor. Peak intensity of the disease process was observed six hours after administration of the inflammatory factor. Protein levels, seromucoid, sialic acid and proteolytic activity in the aqueous humor were correlated with intensity of clinical symptoms observed with the slit lamp. Carrageenin-induced inflammation of the anterior chamber of the eye is an entirely local reaction since the blood serum showed no significant abnormalities of the parameters that were studied. This model, based on biochemical and biomicroscopic studies, may prove useful in the studies of the dynamics of inflammation of the anterior chamber of the eye and for evaluation of anti-inflammatory drugs used in ophthalmology.

Animals

Comparison of the effect of various antisera and cobra venom factor on inflammatory reactions in guinea-pig skin. I. Non-specific inflammation due to the intradermal injection of turpentine.

Guinea-pigs were treated with agents which lowered the serum complement levels and the effect on the inflammatory reaction to intradermal turpentine was studied. A comparison was made between two antisera to beta1C/beta1A globulin which had been prepared in different ways. One was made using C3 fixed to zymosan particles as the antigen, and the other using chemically extracted beta 1C/beta1A globulin. The antisera to beta1C/beta1A globulin (zymosan) was found to have the greater anti-inflammatory activity. The effect of each on the parameters of complement studied, and on the circulating platelet count, was much the same. An antiserum to serum factors fixed to zymosan during C3 activication was prepared free of anti-beta1C/beta1A globulin antibodies. This was found to have some of the anti-inflammatory activity of the anti-beta1C/beta1A globulin (zymosan), as well as lowering total complement levels. Anti-gamma globulin andheterologous immune complexes also lowered serum complement levels and were antiinflammatory. However, all the above agents were also found to reduce the number of circulating platelets. This was in contrast to CVF which reduced complement levels by 90 per cent. but had no effect on platelet numbers. CVF also reduced the inflammatory response but was no more effective than the other agents. In view of these findings a role for the participation of the complement system, possibly involving the alternate pathway of activation of C3, is discussed as well as the participation of platelets in the reaction.

Animals