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MER-BCG (NSC-143769): immunogenicity and toxicity of single and repeated intradermal injections in dogs.

Intradermal injections of MER-BCG 0.1 mg or 0.2 mg at each of 10 multiple sites, led to local granuloma formation. The nodules reached approximately 10 mm in diameter, ulcerated and were accompanied by granulomatous changes in the regional lymph nodes. Six or twelve successive treatments (each including 10 injections) at 4 week intervals produced the same histopathological lesions but no changes in hematological and blood chemical parameters or general morphology and no changes in general condition with exception of occasional weight loss in a few animals. Injection with 0.01 or 0.001 mg/site produced similar, though less severe, skin lesions but no changes in the draining lymph nodes. The immunogenicity of MER-BCG was characterized by granuloma formation, a positive skin response to old tuberculin, and a positive lymphocyte transformation to PPD tuberculin, thus indicating stimulation of cell-mediated immune responses. However, there was a decreased responsiveness to PHA and PPD with continuing treatment with MER-BCG. The decreased responsiveness and accumulation of numerous depots of antigen would suggest an "immunologic paralysis" contraindicating the administration of excessive amounts of MER-BCG during immunotherapy. A specific humoral response to the administration of MER-BCG was not detected, but an MER-BCG dose independent decrease in albumin associated with a non-specific, dose related elevation in serum IgG was observed.

Adjuvants, Immunologic

Cushing's syndrome associated with the intradermal injection of triamcinolone diacetate.

A case of long-term Cushing's syndrome after intradermal injection of 160 mg of triamcinolone diacetate has been presented. The evidence of whether this decrease occurred secondary to the intradermal injections of triamcinolone diacetate or to exogenous abuse is not clear. However, because of the extensive use of intradermal steroids in the treatment of keloids as well as other lesions, this case is significant, whatever the cause. Further study is being accomplished and the final results will be reported by the endocrinology service, Wilford Hall Medical Center, in a follow-up article.

Adolescent

Skin blood flow after intradermal injection of ropivacaine in various concentrations with and without epinephrine evaluated by laser Doppler flowmetry.

BACKGROUND AND OBJECTIVES: Skin blood flow changes after intradermal injection of ropivacaine in various concentrations with or without epinephrine were investigated using laser Doppler flowmetry. METHODS: Twenty-three non-smoking, healthy, young male volunteers participated. Four test sites were used on each forearm (volar surface) in a randomized, double-blind study. Recordings were made at 20, 40, 60, and 90 minutes after intradermal injection (0.1 ml, 30-gauge needle). Injections of saline and 1% lidocaine and an untreated area served as controls. In Series 1, various concentrations of ropivacaine (1%, 0.5%, 0.375%, 0.125%, and 0.063%) were injected. RESULTS: The data from this series showed a dose-response relationship: 1% ropivacaine provoked an increase in skin blood flow similar to saline; 0.5% and weaker concentrations of ropivacaine showed a reduction in flow compared to saline, this being more pronounced with the weakest solutions (0.125% and 0.063%). In Series 2, injection of 1:200,000 (5 micrograms/ml) epinephrine alone and injections of ropivacaine in various concentrations (1%, 0.5%, and 0.25%) with addition of epinephrine were carried out. Injection of epinephrine alone showed a flow almost as low as at the untreated control sites. Ropivacaine epinephrine injections were followed by a lower skin blood flow compared to saline, but the flow was significantly larger compared to the effect of epinephrine itself at the 20-minute recording. CONCLUSION: The combination of ropivacaine and adrenaline did not accentuate but instead diminished the vasoconstrictive effect of epinephrine.

Adult

Pathological changes in calves injected intradermally with Mycobacterium intracellulare serotype Davis and M. avium serotype 2.

Three strains of the M. intracellulare-M. avium complex were injected intradermally into 12 calves. M. intracellulare serotype Davis (M. avium complex serotype 8) was injected into five calves. One calf died with disseminated caseous granulomas at 79 days; three calves killed between 56 and 82 days after inoculation had widely disseminated caseous or caseous or caseocalcareous granulomas. One calf killed at 173 days had only encapsulated granulomas at the injection site and its regional lymph node. Two strains of M. avium serotype 2 (M. avium complex serotype 2) were injected intradermally into seven calves. Six had disseminated granulomas when killed 54-170 days after inoculation. One calf killed at 55 days had granulomas confined to the inoculation sites and their regional lymph nodes. Lesions induced by both M. avium serotype 2 and M. intracellulare serotype Davis were initially caseocalcareous granulomas which became encapsulated and nonprogressive after about 112 days. Two calves inoculated with similar doses of M. bovis and killed in extremis at 37 and 65 days after inoculation had disseminated progressive lesions without encapsulation.

Animals

Epidermal hyperplasia induced in guinea pig flank skin by intradermal injection of Sudan red.

Studies of dermal-epidermal interactions were conducted with guinea pig flank skin and intradermal injections of the irritant, Sudan IV dye in olive oil. These injections led to epidermal hyperplasia in areas overlying the irritant and the effect was most significant when the irritant was placed in the upper dermis. Basal cell mitotic activity and thymidine uptake reached a peak by 24 h and thereafter dropped rapidly. Maximal epidermal thickness (4.3 times the control) resulting from an increase in cell number occurred within 2-4 days. Despite the very short period of increased cell growth, epidermal thickness returned to control values only after a 24-day period. A similar growth response could not be induced by saline injections. A single topical application of the irritant showed a qualitatively and quantitatively different epidermal response. These experiments indicate that an intradermal irritant can lead to epidermal hyperplasia and a long-lasting epidermal thickening.

Animals

Mechanism of local skin atrophy caused by intradermally injected corticosteroids.

Different hydrocortisone microcrystal suspensions applied intradermally in rabbits caused skin atrophy. This could be ascribed to secondary tissue degeneration as was ascertained histologically. The degree of degeneration was related to the concentration of the suspension, size of the microcrystals and frequency of the intervals between the injections. The ground substance of the connective tissue changes and is characterized by a basophilic granular mass, homogenized collagen fibers and degeneration of the elastic fibers. The pathomechanism of local skin atrophy not only differs from connective tissue degeneration caused by systemic corticosteroid treatment but also from the degeneration caused by topical treatment.

Animals

Effect of repeated intradermal injections of bovine herpesvirus type 1 antigen on seronegative cattle.

Forty-three cattle seronegative to bovine herpesvirus-1 (BHV-1) were given from one to five intradermal injections of BHV-1 inactivated antigen at four-week intervals. This delayed hypersensitivity test was assessed by the increase in skin thickness. The activity of the antigen was assessed in five animals which had a previous natural BHV-1 infection with clinical signs and seroconversion. Anti-BHV-1 antibodies were detected by seroneutralisation and an enzyme-linked immunoassay. Only one animal showed a significant but slight increase in skin thickness after the first test, but it was negative after a second test. The animals remained seronegative after the first test. Seroconversion was identified in 11 of the 43 animals (25 per cent) submitted to repeated delayed hypersensitivity tests. Five of 37 animals seroconverted after only two tests. The serological response was transient in seven of 11 seroconverted calves. Repeated hypersensitivity tests were therefore able to induce a serological response in seronegative calves but the response was weak and often transient. The test must therefore be applied cautiously to seronegative animals.

Animals

Prevention of tumor growth after intradermal injection of BCG extracts: a comparison of results in strain-2 guinea pigs from the National Institutes of Health and from the National Jewish Hospital and Research Center.

Line 10, a transplantable hepatocellular carcinoma, was obtained originally from an NIH strain-2 male guinea pig fed diethylnitrosamine. The antitumor activity of BCG and BCG extracts was evaluated in strain-2 guinea pigs obtained both from NIH and the National Jewish Hospital and Research Center (NJH). Animals were immunized with these materials and then tested for their capacity to resist the growth of intradermally injected line-10 tumor cells. Tumor growth was not prevented in 18 NIH animals immunized with living BCG. No tumor growth occurred in 1 of 22 NIH animals immunized with a residue that remained after exhaustive methanol extraction of BCG and in 1 of 44 NIH guinea pigs immunized with BCG extracts. In contrast, tumor growth was prevented in 13 of 22 similarly immunized NJH guinea pigs.

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

[Local reaction to the intradermal injection of PHA in healthy persons and in chronic lympholeukosis patients].

Phytohemagglutinin (PHA) that stimulates the lymphocytes to blast transformation in conditions of cellular cultures provokes local skin reaction, when applied intradermally. At the beginning that reaction is manifested with hypermia at the place of the application and later an infiltration is formed. In healthy subjects the infliltration reaches maximal dimensions on the 48th to the 72nd hour post PHA injection. The presence of lymphocytes was established with the cytologic examination of the material from that infilitration at the 24th hour, lymphoblasts and hyperbasophilic blast cells at the 48th and 72nd hour and single reactive lymphocytes were observed at the 96th hour. The infiltration appears later in patients with quiet development of chronic leukosis, only after the 96th hour and is with smaller demensions. Negligible infiltration is observed in malignant leukosis, its progress in time does not differ from that in healthy subjects. However, skin reaction in intraderma injection of PHA gives a certain orientation about the immune organism reactivity and in case of 1 chronic leaukosis--about the form and severity of the disease.

Adult

Ultrastructural features of epithelioid cell granuloma induced by intradermal injection of xenogeneic nerve tissue.

Epithelioid cell granulomas were induced in rabbits previously sensitised with human sensory peripheral nerve extract by skin testing with homogenate of sural nerve. Ultrastructurally some of the cells contained in their cytoplasm abundant and dilated rough endoplasmic reticulum filled with a moderately dense product while the cytoplasm of other cells contained numerous membrane-bound vesicles. These cells show all the ultrastructural characteristics of epithelioid cells found in human granulomatous disease and in human states of granulomatous hypersensitivity. Thus we have developed an animal model which supports the concept of granulomatous hypersensitivity as a distinct entity in humans to be differentiated from foreign body and delayed-type hypersensitivity reactions. The model may also prove important in elucidating the pathogenesis of granuloma formation in non-lepromatous leprosy and other granulomatous disease and in defining the nature of the products of epithelioid cells.

Animals