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PubMed1984

Oxygen as an antibiotic. The effect of inspired oxygen on infection.

Granulocytes' in vitro bactericidal capacity for certain bacteria depends on an adequate environmental oxygen supply. Oxygen available to granulocytes in infected tissue is decreased by local conditions and can be altered significantly by small changes in its inspired concentration. We modified Burke's and Miles' bacteria-injection model in animals to test the effect of breathing 12%, 21%, and 45% oxygen on the size of lesions produced by intradermal injections of Escherichia coli. Moderately increased inspired oxygen concentrations (fraction of inspired oxygen [Flo2]) significantly decreased the size and number of necrotic lesions, whereas hypoxia increased both. Increasing Flo2 to 45% after three and 24 hours of hypoxia also significantly decreased lesion size. Suppression of infection by moderate hyperoxia is comparable with that reported by Burke after timely, adequate doses of type-specific antibiotics.

Animals↗
PubMed1997

Cyclophosphamide-induced blood and tissue eosinophilia in contact sensitivity: mechanism of hapten-induced eosinophil recruitment into the skin.

The mechanism leading to selective production and accumulation of eosinophils in certain allergic skin diseases is unknown. Cyclophosphamide treatment (150 mg/kg) of BALB/c mice 48 h before sensitization with picryl chloride (PCl) resulted in striking blood and tissue eosinophilia, maximal at 13 days. Blood eosinophilia was not induced by the sensitization with oxazolone and 2,4-dinitrofluorobenzene. Challenge with 1 % PCl, but not croton oil caused preferential eosinophil accumulation into the dermis, which was associated with the enhanced expression of vascular cell adhesion molecule 1 (VCAM-1) on endothelial cells. Intravenous administration of anti-VCAM-1 monoclonal antibody abrogated eosinophil infiltration. In this murine model, we examined the role of several cytokines, including chemokines in inducing selective tissue eosinophilia in vivo. Local administration of antibodies against interleukin (IL)-1beta, IL-4, tumor necrosis factor (TNF)-alpha, and RANTES, but not against IL-5 before challenge inhibited hapten-induced eosinophil recruitment. Intradermal injection of recombinant (r)IL-1beta, rIL-4, rTNF-alpha, rRANTES, and rMIP-1alpha induced marked eosinophil accumulation. Nonetheless, intradermal rIL-5 was not a chemoattractant for eosinophils in vivo. Our findings suggest that IL-1beta, IL-4, TNF-alpha, and RANTES contribute to the selective accumulation of eosinophils in contact sensitivity reaction. Although circulating IL-5 can activate eosinophils and prolong their survival, locally secreted IL-5 is not crucial for inducing eosinophil recruitment into the skin.

Animals↗
PubMed1988

Effect of histamine and prostaglandin E2 on the microcirculation in the skin.

The response of the cutaneous microvasculature to two vasoactive mediators, histamine and prostaglandin E2, was measured by laser Doppler velocimetry. Four concentrations of histamine (1.01 to 65.1 X 10(-5) M, in fourfold dilutions) were injected intradermally into the forearms of six healthy subjects: Prostaglandin E2 (1.77 to 114.0 X 10(-8) M, also in fourfold dilutions) was injected several weeks later. The blood flow at the injection site was measured by laser Doppler velocimetry at five minute intervals until the response resolved. Both mediators produced hyperaemia with markedly accelerated cutaneous blood flow. Analysis of the dose response data showed that, for a given mediator, the differences in responses between subjects were not substantial and, in particular, that the kinetics of the decay of hyperaemia did not vary significantly between subjects. The response to histamine differed from that to Prostaglandin E2 in terms of time to maximum response, duration of maximum response and coefficient of decay.

Adult↗
PubMed1992

Effect of electrostatic forces on the dynamic rheological properties of injectable collagen biomaterials.

Injectable collagen is a concentrated dispersion of phase-separated collagen fibres in aqueous solution used to correct dermal contour defects through intradermal injection. The effect of electrostatic forces on the rheology of injectable collagen was studied by observation of the birefringence of collagen fibres through a polarizing microscope as well as by oscillatory rheological measurements on dispersions of varying ionic strengths (0.06-0.30). The birefringence of fibres progressively increased as ionic strength was reduced from 0.30 to 0.06. The linear viscoelastic measurements displayed a logarithmic relationship between storage (and loss) moduli and frequency over oscillation frequencies of 0.1-100 rad/s. The associated relaxation time spectra, interpreted using the theory of Kamphuis et al. for concentrated dispersions, show that collagen fibres become more flexible as ionic strength increases. This result was analysed at the molecular level from the perspective that collagen fibres are a liquid-crystalline phase of rigid rod collagen molecules which have phase-separated from solution. Electrostatic forces affect the volume fraction of water present in the collagen fibres which in turn alters the rigidity of the fibres. Flexible collagen fibre dispersions displayed emulsion-like flow properties whereas more rigid collagen fibre dispersions displayed suspension-like flow properties. Changes in fibre rigidity significantly alter the injectability of collagen dispersions which is critical in clinical performance.

Animals↗
PubMed2005

The large scale generation of dendritic cells for the immunization of patients with non-small cell lung cancer (NSCLC).

In the current study, we generated large numbers of dendritic cells (DCs) from patients with non-small cell lung cancer (NSCLC) for a vaccine trial. The DCs were generated from CD14+ cells obtained by immuno-magnetic bead column separation technique. The CD14+ cells were placed in culture in the presence of granulocyte macrophage colony stimulating factor (GMCSF) and Interleukin 4 (IL-4). At Day 7, apoptotic bodies derived from an allogeneic NSCLC line 1650-TC were added to the cultures at a DC:tumor cell ratio of 1:1. At Day 8, the DCs were harvested, washed and injected intradermally into patients. Using this protocol we have prepared DCs for 16 patients. An average of 9.3 x 10(7) DCs was injected for the priming dose and 8.2 x 10(7) DCs for the boost. Clinical evaluation of the patients and immune assessment are presented in a separate report. The current report provides evidence for the large scale production of functional DCs derived from patients with NSCLC which can be used as vaccines in clinical trials.

Apoptosis↗
PubMedDate not supplied

Lonomia genus caterpillar toxins: biochemical aspects.

In 1967 we reported for the first time five cases of an acquired bleeding disorder in humans which developed after contact with saturnidae caterpillars. Since that time, other cases have been reported in Brazil, French Guyana, Peru, Paraguay and Argentina. The caterpillars have been identified as Lonomia achelous (LA) in Venezuela and northern Brazil and as Lonomia obliqua (LO) in southern Brazil. All patients present pain and a burning sensation at the site of contact. Within a few hours hematomas and hematuria are seen in combination with intracerebral and intraperitoneal hemorrhage (in some cases also renal failure). Hematological tests show: mild anemia with leucocytosis; prolonged PT, PTT and ThT; decreased fibrinogen, factor V, factor XIII, plasminogen and alpha2-antiplasmin levels; increased factor VIII:c, von Willebrand factor, and FDPs/D-dimers levels with normal ATIII and platelets. Factor VII, factor II and PC levels varied. Several activities similar to or directed against blood clotting factors have been identified in LA: fibrinolytic enzymes, which degrade fibrinogen producing abnormal FDPs; prothrombin activators: one direct and one factor Xa-like; a thermostable factor V activator; a thermolabile factor V inhibitor; a factor XIII proteolytic/urokinase-like activity; and a kallikrein-like activitiy. In LO three activities have been described: a prothrombin activator called 'Lonomia obliqua prothrombin activator protease' (LOPAP); a factor X activator; and a phospholipase A(2)-like activity called Lonomiatoxin. No fibrinolytic activity has been described in LO. Subcutaneous injection of crude hemolymph and some chromatographic fractions of LA induce a decrease in fibrinogen, plasminogen and factor XIII. Intravenous injection of factor XIII proteolytic/urokinase-like activity induce a dose-dependent thrombolysis with a decrease in plasmatic factor XIII without hemorrhagic manifestations. Intradermal injection of LO bristle extracts in rats and rabbits produce incoagulability whereas intravenous injection of LOPAP induced DIC in mice.

Animals↗
PubMed1990

Dissociation of tissue destruction induced by cytolytic T cells in vivo and cytotoxicity as measured in vitro.

Two forms of local cutaneous graft-versus-host reactions were used to examine the in vivo activity of cytolytic T cells in a large number of antigen systems and mouse strain combinations. In immune lymphocyte transfer reactions (TrRs), CTL were injected intradermally into allogeneic hosts to which they were sensitized; in bystander reactions (ByRs), CTL were mixed with target cells and the mixture injected into hosts syngeneic to the CTL. Both reactions frequently culminate in full-thickness skin destruction. However, CTL highly active in cell-mediated lympholysis assays in vitro sometimes failed to induce significant reactions in vivo, and CTL with negligible CML activity often induced severe, necrotizing lesions. In addition, Clone 58, a non-MHC-specific CD8+ clone that originated from cells extracted from a sponge matrix allograft, lost its CML activity but continued to induce necrotizing TrRs and ByRs. Insofar as these reactions may exemplify the specific (TrR) and nonspecific (ByR) tissue injury that occurs in the rejection process, these findings question the reliability of CML for predicting the ability of CTL to induce the tissue destruction seen in allograft rejection.

Animals↗
PubMed1997

Altered tumor growth and metastasis of a T-cell lymphoma in Timp-1 transgenic mice.

The concept of tumor suppression by tissue inhibitor of metalloproteinases (TIMPs) has evolved primarily from studies of genetically modulated tumor cells. The next step is to focus on the host and assess the protective potential of host TIMP-1 on primary tumor growth and metastasis. We generated two transgenic mouse lines with altered Timp-1 expression in skin and liver: one overexpressed Timp-1 (Timp-1(high)), and the other had antisense RNA-mediated Timp-1 reduction (Timp-1(low)). ESbL-lacZ T-lymphoma cells provided the tumor challenge, as they form primary tumors upon intradermal injection with spontaneous metastasis to liver. Metastases were examined in X-Gal-stained whole-organ mounts. Timp-1 overexpression inhibited intradermal tumor growth and spontaneous metastasis, leading to prolonged survival of the mice. The opposite effects occurred in Timp-1(low) mice, leading to shorter host survival. Experimental metastasis assays showed that Timp-1-compromised livers in Timp-1(low) mice showed at least a doubling of metastatic foci and numerous additional micrometastases, indicative of increased host susceptibility. However, Timp-1(high) mouse livers showed an unaltered metastatic load in the experimental metastasis assay. In conclusion, these data demonstrate that Timp-1 levels within a tissue predetermine the development and progression of T-cell lymphoma.

Animals↗
PubMed1986

Contrast-enhanced MRI of periarticular soft-tissue changes in experimental arthritis of the rat.

This study was to determine if manipulation of magnetic resonance signal intensity by means of an intravenously injected paramagnetic contrast agent is useful for the detection and characterization of periarticular inflammation. Arthritis was induced in 20 rats by means of intradermal injection of Freund's complete adjuvant. MR imaging was performed with a resistive magnet operating at 0.35 T. A double spin-echo technique with TE's of 28 and 56 ms and TR's of 0.5 and 2.0 s was used. The hindpaws of the adjuvant-injected rats were imaged on Day 8, Day 11, or Day 15 following injection of the adjuvant. The images were obtained in the transverse plane before and after intravenous injection of gadolinium-DTPA (0.2 mmol/kg). Because of their long T2 relaxation time, inflammatory lesions were characterized by high MR signal intensity on precontrast images obtained with long TR and long TE (T2-weighted images). On the other hand, because of their long T1 relaxation time, the inflammatory lesions were of relatively low intensity and not easily recognized on precontrast images obtained with short TR and short TE (T1-weighted images). Postcontrast T1-weighted images were also sensitive in detecting periarticular inflammation as a result of T1 shortening by the gadolinium-DTPA. However, in our particular model, the data did not indicate any greater MR sensitivity for detecting arthritis by means of gadolinium-DTPA enhancement.

Animals↗
PubMed1984

Evidence by in vivo and in vitro studies that binding of pycnogenols to elastin affects its rate of degradation by elastases.

Procyanidol oligomers and (+) catechin bound to insoluble elastin markedly affect its rate of degradation by elastases. Insoluble elastin pretreated with procyanidol oligomers (PCO) was resistant to the hydrolysis induced by both porcine pancreatic and human leukocyte elastases. The quantitative adsorption of pancreatic elastase was similar on either untreated or PCO-treated elastin suggesting that the binding of this compound to elastin increases the non-productive catalytic sites of elastase molecules. (+) Catechin-insoluble elastin complexes were partially resistant to the degradation induced by human leukocyte elastase but were hydrolysed at the same rate as untreated samples by a constant amount of pancreatic elastase. In addition, the coacervation profile of kappa-elastin peptides as a function of temperature is greatly modified in presence of these flavonoids. We conclusively evidenced that PCOs bind to skin elastic fibres when injected intradermally into young rabbits. As a result, these elastic fibres were found more resistant to the hydrolytic action of porcine pancreatic elastase when injected to the same site. These in vivo studies further emphasized the potential effect of these compounds in preventing elastin degradation by elastase(s) as occurred in inflammatory processes.

Animals↗
PubMed2006

Intradermal positivity after negative prick testing for inhalants.

OBJECTIVE: Examine the incidence of positive intradermal tests after a negative skin prick test for 24 inhalant antigens. STUDY DESIGN AND SETTING: Retrospective study. Charts from patients who underwent modified quantitative testing (MQT) over a 3-year period were reviewed. Patients were initially tested with Multi-Test II. Subjects with negative wheals to a specific allergen were then tested with a 1:500 weight:volume intradermal injection of that allergen. RESULTS: One hundred thirty-three patients underwent MQT for 24 antigens. Allergens with the highest incidence of positive intradermal wheals after negative prick testing were Dermatophagoides pteronyssinus and Dermatophagoides farinae at 26.67%. Allergens with elevated incidence of positive intradermals included fusarium, cockroach, cocklebur, rough marsh elder, and ragweed, all with incidences of 16% to 19%. CONCLUSION: Positive intradermal responses after negative prick testing occur commonly. Future study is needed to determine the generalizability and clinical significance of these findings. EBM RATING: C-4.

Algorithms↗
PubMed1995

Viral interleukin 10 (IL-10), the human herpes virus 4 cellular IL-10 homologue, induces local anergy to allogeneic and syngeneic tumors.

After the cloning of murine cytokine synthesis inhibitory factor, it was recognized that a homologous open reading frame was encoded within the Epstein-Barr virus (human herpes virus 4). This viral protein has now been termed viral interleukin 10 (vIL-10) to reflect its protein sequence homology to "cellular" IL-10 (cIL-10, either murine or human IL-10). It is now widely accepted that vIL-10 shares many functions with cIL-10, principally, the ability to enhance survival of newly infected B cells and to diminish the production of IFN-gamma and IL-2 during ongoing immune reactions. The immunomodulatory effect of locally secreted vIL-10 and murine IL-10 (mIL-10) was examined in tumor models using CL8-1 (a BL6 melanoma cell line transfected with the H-2Kb class I gene) in syngeneic animals. Although parental BL6 tumor cells grow in immunocompetent syngeneic hosts, CL8-1 are rejected. To achieve local secretion of vIL-10, we generated vIL-10 retroviral vectors. While nontransduced CL8-1 cells (1 x 10(4)) failed to grow when injected intradermally in C57BL/6 mice, CL8-1 cells (1 x 10(4)) transduced with vIL-10 formed palpable tumors and eventually killed 80% of injected animals. Suppression of tumor rejection was also noted when CL8-1 tumors with or without vIL-10 transfection were admixed with syngeneic vIL-10-transfected fibroblasts and inoculated. Since the in vitro proliferation of the tumor was not altered after transduction with the vIL-10 gene and injection of vIL-10-transduced CL8-1 does not affect the rejection of nontransduced CL8-1 inoculated at a distant site, local vIL-10 secretion appears to suppress the process of immune rejection of the target cells in a dose-dependent manner. Similar results were observed for the H-2b MCA105 sarcoma tumor model in allogeneic BALB/c mice (H-2d). Although all animals that received nontransfected MCA105 rapidly rejected these tumors, MCA105 sarcomas transfected with vIL-10 remained palpable for up to 37 d. The local immunosuppressive effect of gene-delivered vIL-10 could be neutralized by anti-human IL-10 monoclonal antibody or could be reversed by the systemic administration of IL-2 or IL-12. In marked contrast, mIL-10 transfection of CL8-1 significantly suppressed tumor growth and frequently led to the rejection of tumor. Similar results were obtained for the murine tumor cell lines MCA102.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1981

Influence of PUVA and UVB radiation on delayed hypersensitivity in the guinea pig.

Exposure of guinea pigs to UVA (320--400 nm) radiation following administration of 8-methoxypsoralen by gavage (referred to by the acronym, PUVA) or exposure to UVB (290--320 nm) radiation, produced suppression of the cutaneous delayed hypersensitivity reaction at the site of exposure to radiation and at distant nonexposed sites. In these experiments, the animals were immunized by injection of dinitrophenyl-bovine gamma-globulin (DNP-BGG) in complete Freund's adjuvant and delayed hypersensitivity responses were provoked by intradermal injections of DNP-BGG, DNP and BGG on the flanks. Exposure to erythemogenic doses of either PUVA or UVB radiation for 7 days prior to immunization and for the 7 days between immunization and challenge (total period of radiation: 14 days) produced inhibiton of responses to each of the test substances. In addition, treatment with erythemogenic doses of PUVA either for 7 days prior to immunization or during the interval between immunization and challenge with DNP-BGG, inhibited the delayed hypersensitivity responses at the site of irradiation and at a nonexposed site. These findings suggest that in vivo exposure to nonionizing radiation leads to both local and systemic alteration of certain immune responses.

Animals↗
PubMed1990

Major lymph nodes of the head of the fallow deer (Dama dama) and lymphatic drainage of antlers.

Heads from 10 fallow deer bucks were examined to provide a description of the major lymph nodes in this region. The distribution and size of these nodes were similar to those of sheep and goats. To determine whether there was drainage of lymph from the skin of antlers, and to follow the route of this drainage, a solution containing Evans blue was injected intradermally into the antlers of 2 bucks whilst the animals were anaesthetised. Dye appeared in tracheal lymph ducts 14 to 30 min after injection. The spread of blue colouration in lymphatic ducts and nodes, seen at post-mortem examination, indicated that lymph flowed from antlers laterally into the ipsilateral parotid lymph nodes and from these via medial and lateral retropharyngeal lymph nodes to the tracheal ducts.

Animals↗
PubMed1993

Cutaneous histamine reactivity, histamine content of commercial allergens, and potential for false-positive skin test reactions in dogs.

The cutaneous reactivity of normal and atopic dogs to intradermal injections of histamine phosphate was evaluated. Significant differences were not found in the mean wheal diameters of either group. Commercial allergens used for intradermal skin testing and immunotherapy were determined to contain histamine. To determine whether allergen histamine content was sufficient to cause false-positive skin test results, the cutaneous response of Johnson grass allergic dogs was compared, using commercial Johnson grass allergen and commercial Johnson grass allergen with histamine removed. Significant differences were not noticed between Johnson grass and dehistaminized Johnson grass. Therefore, the histamine content of commercial Johnson grass allergen did not appear to cause false-positive skin test results for this group of Johnson grass allergic dogs.

Allergens↗
PubMed2005

Intradermal botulinum A toxin injection for axillary hyperhydrosis.

Axillary hyperhydrosis in children and teenagers may be severe enough to affect social development. Current treatments range from aluminum chloride compounds to surgical removal of sweat glands and sympathectomy. Intradermal injection of botulinum A toxin has recently been found to be a safe and effective alternative in the adult population. Use in children has so far not been documented. We hereby report a case illustrating the use of botulinum A toxin in a 14-year-old girl with disabling axillary hyperhydrosis.

Adolescent↗
PubMed2000

Analysis of adrenocortical secretory responses during acute an prolonged immune stimulation in inflammation-susceptible and -resistant rat strains.

Endogenous corticosterone secreted during immune challenge restricts the inflammatory process and genetic variations in this neuroendocrine-immune dialogue have been suggested to influence an individuals sensitivity to develop chronic inflammatory disorders. We have tested inflammation-susceptible Dark Agouti (DA) rats and resistant, MHC-identical, PVG.1AV1 rats for their abilities to secrete corticosterone in response to acute challenge with bacterial lipopolysaccharide (LPS) or a prolonged activation of the nonspecific immune system with arthritogenic yeast beta-glucan. Intravenous injection of LPS triggered equipotent secretion of corticosterone in both rat strains. Interestingly, peak concentrations of corticosterone did not differ significantly between the strains. Intradermal injection of beta-glucan caused severe, monophasic, polyarthritis in DA rats while PVG.1AV1 responded with significantly milder joint inflammation. Importantly, serial sampling of plasma from glucan-injected DA and PVG.1AV1 rats did not reveal elevated concentrations of plasma corticosterone at any time from days 1-30 postinjection compared to preinjection values, in spite of the ongoing inflammatory process. Interestingly, adrenalectomized, beta-glucan-challenged DA rats responded with an aggravated arthritic process, indicating an anti-inflammatory role for the basal levels of corticosterone that were detected in intact DA rats challenged with beta-glucan. Moreover, substitution with subcutaneous corticosterone-secreting pellets, yielding moderate stress-levels, significantly attenuated the arthritic response. In contrast, adrenalectomized and glucan-challenged PVG.1AV1 rats did not respond with an elevated arthritic response, suggesting that these rats contain the arthritic process via corticosterone-independent mechanisms. In conclusion, the hypothalamic-pituitary-adrenal axis in both rat strains exhibited strong activation after challenge with LPS. This contrasted to the basal corticosterone levels observed strains during a prolonged arthritic process. No correlation between ability to secrete corticosterone and susceptibility to inflammation could be demonstrated. Basal levels of endogenous corticosterone appeared to restrain inflammation in beta-glucan-challenged DA rats whereas resistance to inflammation in PVG.1AV1 rats may be mediated via corticosterone-independent mechanisms.

Adrenal Cortex↗
PubMed1989

The neutrophil-activating protein (NAP-1) is also chemotactic for T lymphocytes.

T lymphocyte chemotactic factor (TCF) was purified to homogeneity from the conditioned media of phytohemagglutinin-stimulated human blood mononuclear leukocytes by a sequence of chromatography procedures. The amino-terminal amino acid sequence of the purified TCF showed identity with neutrophil-activating protein (NAP-1). Both TCF and recombinant NAP-1 (rNAP-1) were chemotactic for neutrophils and T lymphocytes in vitro supporting the identity of TCF with NAP-1. Injection of rNAP-1 into lymphatic drainage areas of lymph nodes in Fisher rats caused accelerated emigration of only lymphocytes in high endothelial venules. Intradermal injection of rNAP-1 caused dose-dependent accumulation of neutrophils and lymphocytes.

Animals↗