An experimental model of xanthoma by intradermal dextran sulfate injection.
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We have developed in the guinea pig, an animal model of bronchial hyperreactivity provoked by inhalation of trimellitic anhydride (TMA) dust, a known cause of occupational asthma in humans, after intradermal sensitization to the free hapten. Male Dunkin-Hartley guinea pigs (n = 6) were injected intradermally with 0.1 ml of 30% TMA in corn oil. Control animals (n = 7) were injected with 0.1 ml corn oil alone. On Days 21 to 28 after sensitization, guinea pigs were challenged (nose only) to 12 mg/m3 of inhalable TMA dust for 30 min. Bronchial reactivity was measured in sensitized animals and in control animals at 8 h after exposure to the dust. We also measured bronchial reactivity in sensitized exposed guinea pigs at 2 h (n = 5) and at 24 h (n = 5). Pulmonary inflation pressure (PIP) was used to assess bronchopulmonary response. Blood samples were taken for assessment of IgG-1 antibodies to TMA conjugated to guinea-pig albumin. The concentration of acetylcholine required to induce a 100% increase in PIP was used to assess bronchial reactivity. The lungs were eviscerated for histologic examination. All guinea pigs injected intradermally with TMA had high titers of specific IgG-1 antibodies to TMA conjugated to guinea-pig albumin. There was a significant increase in bronchial reactivity in sensitized guinea pigs 8 h after exposure to the TMA dust compared with that in the control animals. There was also a significant eosinophilic inflammatory influx in the subepithelium of the sensitized groups.(ABSTRACT TRUNCATED AT 250 WORDS)
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Chemokines may control mast cell infiltrates found in many inflammatory diseases. These cells act through at least two main functions: migration and degranulation. Here we show that human recombinant monocyte chemotactic protein (MCP)-1 (10 ng/50 microliters) induces, after 4 h, an inflammatory vascular permeability and cellular extravasation reaction, determined by Evan's blue dye (1% in saline) injected into the tail vein of the rat, when injected intradermally in the rat skin. The blue color accumulating at the sites of injection provides evidence of vascular permeability and cellular extravasation. The colored areas of the skin were then enucleated and immersed in a fixative solution. Slides were prepared with sections of tissue colored with toluldine blue and analyzed under an optical microscope. A significant number of basophilic cells migrated to the injected area where MCP-1 (10 ng/50 microliters) was used compared to the control PBS treatment. Cell recruitment was slightly less than N-formyl-methionine-leucyl-phenylalanine (used at 10(-6) M/50 microliters). Electron microscopy studies confirmed the presence of basophilic granular cells where MCP-1 was intradermally injected. After preparation of a histidine decarboxylase (HDC) probe, a Northern blot analysis was determined for HDC mRNA in the enucleated tissue injected with MCP-1 (10 ng/50 microliters). Steady-state levels of HDC mRNA levels were induced after 4 h. These results were confirmed by the higher amount of histamine release, compared to the control PBS, in the enucleated tissue from the MCP-1 injection sites. Our results suggest that MCP-1 could play a significant role in diseases characterized by basophilic cell accumulation and migration to sites of tissue damage. Moreover, we show for the first time that MCP-1 is a pro-inflammatory chemokine that induces basophilic cell migration in rat skin injection sites.
To determine the effects of warming and buffering of 0.5% bupivacaine on the pain associated with intradermal injection and the time of onset of anesthesia, 40 adult volunteers were entered into a randomized, double-blind study conducted at a community teaching hospital. The three-part study compared room temperature (20 degrees) bupivacaine buffered to a pH of 7.1 with the following solutions: buffered bupivacaine warmed to 37 degrees C, unbuffered bupivacaine at room temperature, and unbuffered bupivacaine warmed to 37 degrees C. The same crossover protocol was followed for each part of the study. Subjects received 0.5-mL intradermal injections through 27-gauge needles over 30 seconds, one study solution in each forearm. Immediately after each injection, pain was assessed using a 100-mm visual analog pain scale. The time of onset of anesthesia (loss of intradermal sensation to pinprick) was measured by stopwatch. The mean perceived pain score for the warm buffered bupivacaine (51 mm) was significantly lower than for the room temperature buffered solution (63 mm, P = .003). Similarly, there was a statistical difference between the room temperature buffered and unbuffered solutions (65 v 78 mm, P < .001). The differences in mean pain scores for the room temperature buffered bupivacaine, compared with the other three solutions, suggest that warming and buffering have an additive effect. In this model, the latency of action of bupivacaine was not affected by alkalinization. However, warming bupivacaine to 37 degrees C reduced the time of onset to intradermal anesthesia by 12.1 seconds (95% confidence interval, 0.6 to 23.6). These results suggest that warming is more effective than buffering to reduce the pain of infiltration of bupivacaine and the time of onset of intradermal anesthesia.
High molecular weight (19S) gamma-globulin produces passive cutaneous anaphylaxis (PCA) reactions with its specific antiserum only when it is used as intravenous antigen. Reversed PCA reactions cannot be produced when the 19S protein is injected intradermally in contradistinction to the results with 7S gamma-globulin. Antibodies of the high molecular weight class, when injected intradermally, also failed to give PCA reactions following antigen injection. Heterophile antibodies from the sera of patients with infectious mononucleosis, demonstrated to be entirely of the 19S type, gave negative reactions, while 7S heterophile antibodies from guinea pig did give reactions following intravenous injection of sheep cells. Anti-B isoagglutinins of the 19S class failed to react, while those of the 7S class of similar titer gave clear reactions following the injection of B substance. Evidence was obtained that 19S antibodies were capable of inhibiting the PCA reactions obtained with the 7S type, indicating that interaction with antigen occurred. The failure to elicit PCA reactions appeared to be due to an inability to fix to guinea pig tissues in a manner similar to that known for 7S antibodies and gamma-globulin.
Twenty-two centers participated in a study to determine the efficacy and safety of gelatin matrix implant (GMI; Fibrel) in the elevation of depressed cutaneous scars. Gelatin matrix implant is an implant consisting of absorbable gelatin powder and epsilon-aminocaproic acid, which is reconstituted with the patient's plasma before being injected intradermally beneath the scar. A total of 321 patients were evaluated after a skin sensitivity test; six of the patients (1.9%) had a positive response. After a skin test with negative results, 27 patients dropped out of the study for unrelated reasons. The remaining 288 patients were treated with the implant (total of 840 scars), many of whom have been followed up for more than 1 year. Preliminary results show that approximately half of the treated scars showed improvement of greater than 65%. The current data suggest that the improvement lasts at least up to 1 year. Adverse reactions to gelatin matrix implant injections were local and transient, and none of the patients developed major hypersensitivity responses to the treatment. The data indicate that intradermal injections of gelatin matrix implant are safe and effective in correcting the depressed scars selected for this study.
A total of 48 patients with gastric cancer were randomly assigned to receive either endoscopic injections of OK-432 plus systemic treatment with intravenously administered 5-fluorouracil and intradermally injected OK-432 (group A) or systemic therapy alone (group B). Morphologic improvement occurred in 7 of the 22 patients in group A. In one patient the tumor mass disappeared completely. In group B morphologic improvement occurred in only 2 of the 26 patients. The survival rate throughout the 24-month period was significantly (P less than 0.01) higher in group A than in group B patients. Thus, local administration of OK-432 by endoscopic injection is effective and is recommended for the treatment of patients with advanced gastric cancer.
OBJECTIVE: To examine the effect of listening to self-selected music versus an intradermal injection of normal saline solution on the intensity and distress of intravenous (IV) catheter insertion pain. DESIGN: Prospective, randomized, controlled study. SETTING: Inpatient and outpatient units in 2 university-affiliated southwestern medical centers. PATIENTS: One hundred ten adult inpatients and outpatients undergoing IV therapy. OUTCOME MEASURES: Pain intensity, pain distress, and IV insertion difficulty visual analog scales. INTERVENTION: Patients were randomly assigned to receive an intradermal injection of normal saline solution, listen with headphones to self-selected music, or be in a control group for IV insertion. RESULTS: A MANOVA revealed no statistically significant multivariate or univariate differences in pain by treatment group, but significantly higher pain distress scores with failed IV insertions. The pain intensity and distress scores were significantly higher in the saline solution group when compared with the music and control groups combined. Insertion difficulty was significantly positively correlated with pain intensity and distress for the entire sample, with weak, nonsignificant correlations in the music group. CONCLUSION: Intradermal unpreserved saline solution contributes to greater pain intensity and distress, greater insertion difficulty, and a higher failure rate than the use of music or routine IV insertion. Listening to preferred music attenuates the effect of insertion difficulty on IV insertion pain. Intravenous insertion attempts were unsuccessful in more than one third of the subjects, resulting in higher pain distress scores. Further research is needed on interventions to reduce IV insertion pain and on factors contributing to IV insertion failure.
OBJECTIVE: To examine the effect of topically applied tea tree oil (TTO) on histamine-induced oedema in the ears of mice. METHODS AND RESULTS: For BALB/c mice, 10 microl undiluted TTO applied immediately after, but not 30 min before intradermal injection of 600 microg histamine in 10 microl, significantly suppressed oedema development. TTO applied after histamine injection also suppressed histamine-induced oedema in C57/BL6 mice. TTO applied immediately after intradermal injection of compound 48/80 (200 microg in 10 microl saline) also significantly reduced ear swelling. TTO suppressed histamine-induced oedema to the same extent in capsaicin-treated (neuropeptide-depleted) and control mice which suggests that TTO does not inhibit histamine-induced oedema by regulating the activity of peripheral sensory neurons. Terpinen-4-ol, the major water-soluble component of TTO, was equivalent in potency to TTO in the suppression of histamine-induced ear swelling. CONCLUSION: Topical application of TTO, and in particular terpinen-4-ol, may be effective in controlling histamine-induced oedema often associated with Type I allergic immediate hypersensitivities.
This study has compared the buffering capacity of inflamed skin with that of normal skin, in rats, following the injection of commonly used, relatively acidic local anaesthetic solutions. Inflammation was induced by the intradermal injection of 1% carrageenan 0.1 ml. Local anaesthetic solution (0.05 ml) was injected intradermally into four to six animals in the irritant (inflamed skin) and control (normal skin) groups. Tissue pH changes were measured continuously by a Roche tissue pH electrode for up to a maximum of 180 min. Contrary to the generally accepted view, there was an increase in buffering capacity in inflamed tissues, thus offering no support for the hypothesis that tissue acidity in inflammation is an important factor in the failure of local anaesthetics to act in inflamed tissues.
We have previously shown that inhaled Con A has a powerful enhancing effect on the formation of immune complexes between an inhaled antigen and circulating antibody. Immunohistochemical staining has demonstrated such complexes, together with host complement, in close association with foci of necrotizing destruction of the pulmonary parenchyma. We have postulated that Con A promotes immune complex formation indirectly through polyclonal activation of lymphocytes in the lung. In this paper we test this hypothesis in animals rendered unresponsive to Con A stimulation in vivo by i.v. administration of cholera toxin (CT). Such treatment raised the levels of cAMP in peripheral blood lymphocytes and inhibited their proliferative response to Con A in vitro. CT administration further blocked the local inflammatory response to intradermal injections of Con A, as well as the cell-mediated immune response to intradermal injections of BSA. Although CT failed to block the immune complex-mediated Arthus vasculitis in the skin, it did block production of immune complex pulmonary injury by antigen/mitogen aerosols, as did decomplementation with purified cobra venom factor. These findings support the hypothesis that polyclonal activation of pulmonary lymphocytes promotes immune complex-type alveolitis, possibly by facilitating interactions between humoral antibody and intra-alveolar antigen.
15-Hydroxyeicosatetraenoic acid (15-HETE), a 15-lipoxygenase product of arachidonic acid, inhibits leukotriene B4 (LTB4)-induced chemotaxis of polymorphonuclear leukocytes (PMNs) in vitro. In this study the effects of intradermal injections of LTB4 were determined in the absence or presence of 15-HETE. For comparison intradermal injections of purified human complement split product C5a were performed in the absence or presence of 15-HETE. The skin response was evaluated by measuring the diameter of the wheal, the area of the flare and by intensity of the erythema (erythema index). LTB4 and C5a were injected at the concentration of 200 ng/ml. At this concentration the maximal skin response of LTB4 and C5a were equivalent. In contrast to C5a reaction, which resolved within 1 h, LTB4-induced skin response lasted up to 18 h. In all subjects the skin response was significantly decreased when LTB4 was injected together with 300 ng of 15-HETE. The decrease of wheal, flare, and erythema index averaged 81.9%, 56.6%, 53.6%, respectively, when all parameters were obtained at the maximal skin response. In contrast, the C5a-induced skin response was not affected by addition of 15-HETE, even when the final dose of 15-HETE was increased 10 times to 3 micrograms. The LTB4-induced reaction could last up to 18 h after injection. After the addition of 300 ng of 15-HETE the skin response resolved after 1 h. The present results demonstrate that 15-HETE is a specific inhibitor of the LTB4-induced skin response and brings additional evidence in support of the ability of 15-HETE to regulate the proinflammatory effects of LTB4 in vivo.
The morphological aspect of the antineoplastic defence mechanism was studied in Sewall-Wright strain 13 guinea pigs using isogeneic line 2 hepatoma cells. When these tumour cells were injected i.p. into isogeneic hosts, they grew rapidly and always killed their hosts within 8 weeks. An intradermal injection of line 2 hepatoma cells in strain 13 guinea-pigs 3 weeks before i.p. challenge with the same tumour cells protected such pre-immunized animals. While the intradermal inoculum failed to grow, it conferred sufficient immunity to enable the host to reject the tumour after a short period of growth. This phenomenon showed immunological specificity since intradermal injection of allogeneic hepatoma cells did not offer this protection. By using sequential histological analysis in these preimmunized animals, we were able to identify 2 sets of cells migrating into and around tumour modules in relation to tumour cell death. Firstly, small numbers of small lymphocytes were observed at the earliest signs of tumour cell damage. They were most likely the effectors of antineoplastic defence mechanisms. Secondly, a variety of numerous inflammatory cells including macrophages were observed later when the tumour was necrotic, and these cells were considered to be present as a response to tumour cell death.
To elucidate the mechanism of vacuolation frequently encountered in the normal epidermal cells surrounding skin tumors, 0.5 and 1.0% procaine hydrochloride, and 1.0% lidocaine hydrochloride with and without epinephrine were injected either i.d. or s.c. into mature white male rabbits, and the sites of injection were biopsied either immediately or after 15 min. the epidermis of the site of intradermal injection showed a significant increase in the number of vacuolated cells in contrast to the site of s.c. injection (p less than 0.001) irrespective of the kinds of local anesthetics and the time lapse from injection to biopsy. Electron microscopy revealed vacuole-formations without limiting membrane in the cytoplasm of the keratinocytes. It is suggested that the local edema in the upper dermis caused by injection of anesthetics gives rise to edema and vacuolation of overlying epidermal cells, and that i.d. injection of local anesthetics may be regarded as a cause of epidermal vacuolation.
Central sensitization of spinothalamic tract (STT) neurons in anesthetized monkeys after intradermal injection of capsaicin depends in part on disinhibition. Protein kinase C is suggested to participate in this process. The present study shows that the nitric oxide-cGMP (NO-cGMP) signal transduction system also contributes to sensitization of wide dynamic range (WDR) STT neurons located in the deep dorsal horn. The NO-cGMP system was activated by microdialysis administration into the dorsal horn of 8-bromo-cGMP, an analog of cGMP. Sensitization of STT cells by 8-bromo-cGMP increased the responses of deep WDR STT cells to both weak and strong mechanical stimulation of the skin and simultaneously attenuated the inhibition of the same neurons produced by stimulation in the periaqueductal gray (PAG). In contrast, WDR STT cells in the superficial dorsal horn and high-threshold (HT) STT cells in superficial or deep layers showed reduced responses to mechanical stimulation of the skin after infusion of 8-bromo-cGMP, and PAG inhibition of these neurons was unaffected. Sensitization of STT cells and the attenuation of PAG inhibition induced by intradermal injection of capsaicin were prevented by preteatment of the dorsal horn with a guanylate cyclase inhibitor, 1 H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one. The results support the hypothesis that activation of the NO-cGMP signal transduction system contributes to the sensitization of WDR STT neurons in the deep dorsal horn and helps explain why intradermal capsaicin injections often fail to sensitize superficial and HT STT cells. The results also support the idea that sensitization of STT cells is produced in part by disinhibition.
PURPOSE: The aim of this study is to elucidate the effects of laparoscopic Nissen fundoplication (LNF) with carbon dioxide (CO(2)) or helium (He) on the cell-mediated immune response in a pediatric animal model compared with open Nissen fundoplication (ONF). METHODS: Cell immune response was evaluated in 45 1-week-old Sprague Dawley rats using the delayed type hypersensitivity (DTH) skin test. Animals were sensitized against keyhole limpet hemocyanin (KLH) by subcutaneous injection (0.5 mg) in complete Freund's adjuvant. Animals were challenged 2 weeks later by an intradermal injection of KLH (0.3 mg) in sterile saline (challenge 1, baseline). Rats with positive DTH skin reaction at 24 and 48 hours after challenge 1 were put randomly into 4 groups (n = 10 each): I, only anesthesia (control); II, LNF with CO(2), III, LNF with He; IV, ONF. Animals were injected intradermally with KLH (0.3 mg) immediately before the procedures (challenge 2) and 3 and 6 days postoperatively (challenges 3 and 4). RESULTS: DTH skin reactions were measured 24 and 48 hours after each challenge. There were no significant changes in cell-mediated immunosuppression after LNF with CO(2). However, a transient cell-mediated immunosuppression was observed after LNF with He and ONF. All fundoplications were intact at the time of necropsy. CONCLUSIONS: These data suggest a transient suppression of cell-mediated immunity in open procedures when compared with laparoscopic interventions using CO(2) in a pediatric animal model. In addition, the type of gas used during laparoscopy also may modulate this transient immunosuppression.