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Morphine-induced skin wheals: a possible model for the study of histamine release.

We evaluated the ability of morphine to release histamine when injected intradermally in man. Mathematic analysis of the dose-response (wheal) relationship suggested that two different effects were involved. The effect of low doses of morphine (0.05 to 1 microgram) was clearly antagonized by naloxone (0.4 or 1.2 mg im 30 minutes before), whereas the effect of higher doses (5 to 50 micrograms) was not modified. The median effective doses of morphine (ED50) for the low dose range effect were 0.07 +/- 0.01 and 0.08 +/- 0.01 microgram before naloxone and 0.14 +/- 0.02 and 0.15 +/- 0.03 microgram after 0.4 and 1.2 mg doses, respectively. Astemizole (45 mg po 30 minutes before) and oxatomide (60 mg po 120 minutes before) produced similar inhibition of histamine-induced wheals, but there were clear differences in their effects on wheals elicited by morphine. Morphine ED50 values for the low dose range effect rose from 0.09 +/- 0.01 to 0.20 +/- 0.01 microgram after astemizole and from 0.08 +/- 0.01 to 0.46 +/- 0.04 microgram after oxatomide. Opiate receptors may be involved in some of the effects produced by morphine injection in the human skin, but morphine-induced wheals seem to offer a suitable model for the evaluation of agents capable of inhibiting histamine release in man.

Adult↗

Cutaneous basophil hypersensitivity in atopic dermatitis.

The infiltration by basophils into delayed hypersensitivity skin test sites was examined in patients with atopic dermatitis, contact dermatitis and in normal healthy persons. Atopic dermatitis patients, with large amounts of IgE, injected intradermally with staphylococcal antigens showed reactions that were more transient and erythematous than those of normal persons. On histological examination there were numerous, degranulating basophils among the perivascular mononuclear cells. Normal persons, with small amounts of IgE showed typical mononuclear cell infiltration and few basophils. Patients with contact dermatitis, one with much IgE, responded to patch tests to potassium dichromate or to nickel sulphate by delayed type reactions, but on histology, two of the four patients showed a significant infiltration by basophils. It is considered that the erythematous response to the antigen in atopic dermatitis patients is related to the basophil infiltration.

Adult↗

[Axillary hyperhidrosis: successful treatment with botulinum toxin A].

Severe hyperhidrosis may present as a vexing condition for the patient and a therapeutic challenge for the physician. As botulinum toxin-A (BT-A) can potently block cholinergic sympathic innervation of sweat glands it was used in an open trial for controlling severe axillary hyperhidrosis. After visualization of hyperhidrosis using the iodine-starch-test BT-A (Dysport, 400 Units) was injected intradermally in one axilla. One week later gravimetric quantification of sweat secretion revealed a reduction of sweating to 4-9% of initial values. In order to rule out daily variations as well as other factors interfering with sweat production BT-A treatment was restricted to only one axilla in each individual clearly demonstrating extensive anhidrotic efficacy on the treated side only. The treatment was well tolerated without side-effects and was assessed as 'completely satisfying' by all patients. BT-A may offer a fast, safe and highly effective therapeutic option for severe hyperhidrosis.

Adolescent↗

SR142948A is a potent antagonist of the cardiovascular effects of neurotensin.

The novel compound SR142948A was compared with SR48692 as an antagonist of neurotensin-induced cardiovascular effects both in vitro and in vivo. SR142948A inhibited [125I]-neurotensin binding [median inhibitory concentration (IC50) = 0.24 +/- 0.01 nM], neurotensin-induced cytosolic free Ca2+ increase (IC50 = 19 +/- 6 nM), and prostacyclin production in human umbilical vein endothelial cells (IC50 = 17 +/- 3 nM) at much lower concentrations than did SR48692 (respective IC50 values, 14 +/- 5, 41 +/- 16, and 86 +/- 16 nM). Oral administration of SR142948A (10 microg/kg) resulted in significant inhibition of neurotensin-induced blood pressure changes, whereas SR48692 was active only at 10-fold higher doses. Furthermore, SR142948A administered i.v. in microg/kg quantities in the rat was as active as mg/kg doses of SR48692 on neurotensin-induced increase in hematocrit. SR142948A injected intradermally also significantly inhibited neurotensin-induced plasma extravasation at concentrations as low as 10 pmol/site, whereas 1,000 pmol/site of SR48692 were necessary to reach a significant inhibition. These data show that SR142948A is a novel, extremely potent antagonist of neurotensin-induced cardiovascular responses both in vitro and in vivo. SR142948A and SR48692 constitute a pair of nonpeptide neurotensin antagonists of different potency, which may be used to probe for the implication of neurotensin receptors in physiologic or pathologic phenomena.

Adamantane↗

The exotoxins of Corynebacterium ulcerans.

The exotoxins produced by ten strains of C. ulcerans (two human, six bovine and two equine) have been studied. On the criteria of toxin-antitoxin neutralisation and immunoprecipitation tests using highly specific diphtheria and C. ovis antitoxins with crude toxic filtrates, (NH4)2SO4 concentrates, and partially purified chromatographic preparations of these, together with the presence or absence of inhibition of the action of staphylococcal beta-haemolysin, and the reaction produced when injected intradermally into rabbits, two toxins could be identified, namely diphtheria toxin and C. ovis toxin. There was no evidence for the production of a third toxin specific for C. ulcerans. Five strains produced both diphtheria and C. ovis toxins. In four diphtheria toxin predominated, but in the fifth C. ovis toxin predominated. Two strains produced only diphtheria toxin and two only C. ovis toxin, though there was good but not complete evidence that a third strain (Revell) also fell into this latter group. Considerable variation occurred in the concentration of each toxin and, where both were present, in the proportion of each.

Animals↗

Olvanil: more potent than capsaicin at stimulating the efferent function of sensory nerves.

The capsaicin analogue olvanil stimulated an increase in cutaneous blood flow when injected intradermally into the anaesthetised rabbit, as measured using a 133Xenon clearance method. Olvanil was found to be a 10-fold more potent vasodilator (on a molar basis) than capsaicin. The effect of both vasodilators was significantly inhibited by the calcitonin gene-related peptide (CGRP) receptor antagonist CGRP-(8-37). These findings suggest that olvanil stimulates the efferent function of cutaneous sensory nerves in a more potent manner than capsaicin. We therefore suggest that olvanil is a useful pharmacological tool for studying the activity of neuropeptides released from sensory nerves.

Anesthesia↗

Perception of histamine-induced itch elicited in three different skin regions.

In order to investigate whether any regional difference in itch perception exists in experimentally induced pruritus, various concentrations of histamine (1, 3.3, 10, 33 and 100 micrograms/ml) were injected intradermally in three different skin regions in 15 healthy subjects. The regions were 1) the volar aspect of the forearm, 2) the lateral aspect of the upper arm and 3) the upper back at scapular level. Itch perception (itch latency, itch duration, maximal itch intensity, 'total itch index') and flare reactions were studied. A significant dose-response relationship was shown in each of the three regions for all itch variables, except itch latency, and for the flare reactions. However, no significant regional difference in itch perception was observed. The flare reaction showed a regional variation, with significantly smaller flares on the forearm than in the other two regions.

Adolescent↗

In vitro and in vivo interactions of Haemophilus ducreyi with host phagocytes.

We investigated the phagocytosis of Haemophilus ducreyi both in vitro and in vivo. Human granulocyte and monocyte phagocytosis of opsonized and nonopsonized, fluorescence-labeled H. ducreyi was assessed by flow cytometry. Both Escherichia coli and noncapsulated H. influenzae were included as controls. The maximal percentage of granulocytes taken up by H. ducreyi was 35% after 90 min. In contrast, 95% of H. influenzae bacteria were phagocytosed by granulocytes after 30 min. These results indicated that H. ducreyi phagocytosis was slow and inefficient. Bacterial opsonization by using specific antibodies increased the percentage of granulocytes phagocytosing H. ducreyi from 24 to 49%. The nonphagocytosed bacteria were completely resistant to phagocytosis even when reexposed to granulocytes, indicating that the H. ducreyi culture comprised a mixture of phenotypes. The intracellular survival of H. ducreyi in granulocytes, in monocytes/macrophages, and in a monocyte cell line (THP-1) was quantified after application of gentamicin treatment to kill extracellular bacteria. H. ducreyi survival within phagocytes was poor; approximately 11 and <0.1% of the added bacteria survived intracellularly after 2 and 20 h of incubation, respectively, while no intracellular H. influenzae bacteria were recovered after 2 h of incubation with phagocytes. The role of phagocytes in the development of skin lesions due to H. ducreyi was also studied in vivo. Mice that were depleted of granulocytes and/or monocytes and SCID mice, which lacked T and B cells, were injected intradermally with approximately 10(6) CFU of H. ducreyi. Within 4 days of inoculation, the granulocyte-depleted mice developed lesions that persisted throughout the experimental period. This result reinforces the importance of granulocytes in the early innate defense against H. ducreyi infection. In conclusion, H. ducreyi is insufficiently phagocytosed to achieve complete eradication of the bacteria. Indeed, H. ducreyi has the ability to survive intracellularly for short periods within phagocytic cells in vitro. Since granulocytes play a major role in the innate defense against H. ducreyi infection in vivo, bacterial resistance to phagocytosis probably plays a crucial role in the pathogenesis of chancroid.

Animals↗

Effect of multiple interferon gamma injections on the disposal of Mycobacterium leprae.

The effect of multiple intradermal injections (four to six) of 10 micrograms of interferon gamma on the number of Mycobacterium leprae in the skin of patients with polar lepromatous leprosy and borderline lepromatous leprosy was evaluated. To achieve a maximum zone of induration and cell emigration a preparatory dose of the lymphokine was required. A second group of three injections, given 3-4 days after the initial series, resulted in lesser degrees of induration and was more in keeping with a partial local hyporesponsive state. A marked emigration of T cells and monocytes into the dermis resulted from injections of interferon gamma and persisted for greater than 21 days. A preponderance of CD4+ cells in the infiltrate was seen within a few days and CD4/CD8 ratios remained elevated for greater than 5 weeks. The bacillary load of injected sites evaluated 21 days after lymphokine administration was reduced in 14/17 patients by factors ranging from 5- to 1000-fold. This occurred predominantly within diffuse lesions and occurred rarely in nodular sites. Biopsy samples of injected sites taken 6 months later demonstrated progressive 10-fold reductions in bacilli and the continued presence of a granulomatous response.

Administration, Topical↗

Local effect of vasoactive intestinal polypeptide on human sweat-gland function.

Physiological significance of vasoactive intestinal polypeptide (VIP), a putative co-transmitter of the cholinergic neuron innervating sweat glands, was investigated by its local effect on drug-induced sweating. VIP, methacholine chloride (MCH), or VIP plus MCH dissolved in 0.1 ml of 0.9% NaCl solution to a specified concentration was injected intradermally at the center of a forearm test area of 15 cm2 and the sweat rate was recorded continuously by capacitance hygrometry. In a cool environment (Ta, 23 degrees C), VIP failed to cause sweat secretion, but increased the rate of MCH-induced sweating, most markedly at a concentration of 10(-5) g/ml, where the rise in local skin temperature was the greatest. On an area anesthetized by nerve block in a hot environment (Ta, 35 degrees C), the effect was less obvious and less consistent, indicating that the sweat-facilitatory effect of VIP is reduced under the condition of passive cutaneous vasodilation. It may be postulated that VIP plays a role in securing ample oxygen supply to functioning sweat glands, especially with a relatively high cutaneous vasoconstrictor tone.

Adult↗

[Detection of Bacillus cereus-toxins (author's transl)].

During the logarithmic state of growth, B. cereus produced extracellular toxins, which could be precipitated by ammoniumsulphate. The toxins were not dialysable. 9 strains of B. cereus were tested and showed lethal, hemolytic, and phospholipase-C-reactions. The lethal toxin injected i.v., killed mice within 20 minutes. An intradermal injection of the 9 toxin-precipitates in rabbits produced skin reactions, which could be neutralized by a specific antiserum against the B. cereus strain B-4ac. Relatively high doses (50 mg/ml) of the toxin caused fluid accumulation after injection in ligated loops of rabbit gut (Tab). The hemolysin of the tested B. cereus strains was found to be thermolabile. The phospholipase-C activity withstood the temperature of 55 degrees C for a period of 10 minutes. The prepared specific antiserum, which inhibited the phsopholipase-C activity totally, had no effect on the hemolysis. Intradermal and enteropathogenic activities point to the presence of an enterotoxin. For detection of the enterotoxin, the skin-test in guinea pigs or rabbits seems to be most appropriate.

Ammonium Sulfate↗

A randomized, double-blind, placebo controlled study on analgesic effects of botulinum toxin A.

OBJECTIVE: Botulinum toxin type A (BTXA) is used to treat neurologic disorders associated with increased muscle tone. Its use is often associated with pain relief. METHODS: A possible direct analgesic effect of BTXA on C and Adelta fibers was studied on 16 healthy volunteers receiving 30 U BTXA into one forearm and pure saline into the other. To exclude the secondary effect due to muscular tone reduction, BTXA was injected intradermally. Thermal sensory testing of heat pain (threshold and tolerance) and neuroselective current sensory testing of current pain threshold/tolerance were performed at baseline and 3, 14, and 28 days after treatment. Thereafter, on day 28, capsaicin was administered simultaneously into both forearms to evaluate a possible peripheral effect and central effect on pain processing and on the axon reflex flare. RESULTS: The authors observed no significant difference in any of the perception outcome measures between BTXA and placebo pretreated areas. Flare areas as a result of the release of neuropeptides after capsaicin application showed no differences. CONCLUSIONS: The results suggest that pain reduction after BTXA treatment is mediated through its effect on muscle tone rather than a direct analgesic effect.

Adult↗

[Evaluation of the specific delayed cutaneous hypersensitivity response(DTCH) after specific active immunotherapy (SAI) using autologous tumor cells in patients operated upon for renal carcinoma].

From Dec 1986 to Dec 1990, 113 consecutive patients radically resected for renal cell carcinoma, have been randomized after surgery to observation or to Active Specific Immunotherapy (A.S.I.). 43 patients stage I-II and 13 p. stage III, according to TNM entered the treatment arm consisting of 10 autologous irradiated tumor cells injected intradermally, either mixed with BCG 10(7) (on days 28th and 35th after surgery) or alone (on day 42th). At randomization and 1, 6 and 12 months after treatment, patients were evaluated for the development of a DTCH to autologous tumor and to autologous normal cells, obtained by mechanical and enzymatic dissociation of the surgical specimens. Baseline DTCH were negative in all patients. One month after completing A.S.I. 36 out of 50 evaluable patients displayed a significant DTCH response to autologous tumor, which remained positive in 23/40 patients at 6 months. Our data clearly indicate that Active Specific Immunotherapy with autologous tumor cells mixed with BCG can elicit a specific immune response to autochthonous tumor as measured by DTCH.

Adult↗

Interstitial lymphangiography: initial clinical experience with a dimeric nonionic contrast agent.

Interstitial lymphangiography performed with intradermal injection of a new nonionic dimeric contrast agent was used to examine 34 normal lower extremities and 35 lower extremities with proved lymphedema. An infusion pump was used to inject 2 mL of the contrast agent into the intradermal interdigital space at a rate of 0.1 mL/min. In all normal extremities, the injection deposit appeared homogeneous, and one to five lymph vessels could be identified. Diffusion of contrast agent into perilymphatic tissue occurred 30-40 cm proximal to the injection site; inguinal lymph nodes were, therefore, not routinely visualized. The findings at interstitial lymphangiography were abnormal in all extremities with advanced lymphedema. The results indicate that interstitial lymphangiography with the use of a nonionic dimeric contrast agent allows morphologic assessment of lymphatic vessels and facilitates the diagnosis of lymphatic obstructive disease.

Adolescent↗

The C-terminal domain promotes the hemorrhagic damage caused by Vibrio vulnificus metalloprotease.

Vibrio vulnificus, an opportunistic human pathogen, produces a 45-kDa zinc metalloprotease (V. vulnificus protease; VVP) as an important virulence determinant. VVP injected intradermally into the dorsal skin causes the hemorrhagic damage through specific degradation of type IV collage in the vascular basement membrane. The N-terminal 35-kDa polypeptide (VVP-N), the catalytic domain, also evoked the hemorrhagic skin reaction within minutes. However, the hemorrhagic activity of VVP-N was one-third of that of VVP. Besides, the proteolytic activity of VVP-N toward the reconstituted basement membrane or type IV collagen was found to be about 50 % of VVP. VVP-N, like VVP, was quickly inactivated by an equimolar amount of alpha(2)-macroglobulin, a broad-spectrum plasma protease inhibitor. These findings indicate that the C-terminal 10-kDa polypeptide, the substrate-binding domain mediating the effective binding to protein substrates, functions to augment the hemorrhagic reaction of VVP.

Amino Acid Sequence↗

Sentinel node detection and definition may depend on the imaging agent and timing.

PURPOSE: Two cases of sentinel lymph node imaging are presented in which the results are exceptions to what the literature generally defines as sentinel lymph nodes. In one case, Tc-99m antimony trisulfide colloid produced significantly different results than did Tc-99m tin colloid. In the second case, the results bring into question the definition of a sentinel node as the first in a lymphatic drainage pathway. MATERIALS AND METHODS: In one patient, lymphoscintigraphy was performed initially using Tc-99m antimony trisulfide colloid injected intradermally around a melanoma excision site. Repeated lymphoscintigraphy 1 month later, 1 hour before sentinel node excision, was done using Tc-99m tin colloid, a larger particle than antimony trisulfide colloid. The second patient, with a melanoma biopsied only, had sentinel node imaging performed using Tc-99m sulfide colloid, a particulate also larger than antimony trisulfide colloid and also 1 hour before sentinel node excision. RESULTS: In the first patient, imaging with the smaller antimony trisulfide colloid showed more lymphatic pathways and more sentinel nodes than with tin colloid. In the second patient, the first focus of retention of the imaging agent in the lymphatic pathway seen showed less intense accumulation than the next focus in the pathway, contrary to published reports that the sentinel node shows more intense accumulation than do nodes further downstream in a lymphatic pathway. CONCLUSIONS: There are exceptions to published characteristics of sentinel node lymphoscintigraphy, so care must be exercised in localizing sentinel nodes.

Antimony↗

High-density proteoglycan induces specific suppression of adjuvant-induced arthritis in rats.

In vitro data support the view that T cells in adjuvant-induced arthritis (AIA) respond to the proteoglycan (PG) component of articular cartilage; however, an in vivo role for PG in AIA has yet to be shown. To do so, we examined the effects of pretreatment with bovine cartilage high density PG (HDPG) on AIA induced by heat-killed Mycobacterium butyricum in Lewis rats. Purified bovine cartilage HDPG emulsified in Freund's incomplete adjuvant (FIA) was injected intradermally into rats 7 days before challenge with Myco. butyricum. The severity of arthritis was significantly suppressed in rats pretreated with as little as 0.75 mg of HDPG, and the arthritis was completely suppressed in rats pretreated with 3.0 mg of HDPG. This suppression was specific, as the same treatment did not protect against type II collagen-induced arthritis. Suppression of AIA is primarily a property of the HDPG, as suppression of the arthritis was significantly less with pretreatment with 3.0 mg of middle density fractions of PG, and no suppression was observed with pretreatment with the lowest density fraction of PG. Thus we report that pretreatment with cartilage HDPG, but not lower density PG, can induce specific suppression of AIA. These in vivo results support the view that immunity to cartilage HDPG plays a major role in the pathogenesis of AIA, and can induce specific tolerance to this type of arthritis.

Animals↗

Tea tree oil reduces histamine-induced skin inflammation.

BACKGROUND: Tea tree oil is the essential oil steam-distilled from Melaleuca alternifolia, an Australian native plant. In recent years it has become increasingly popular as an antimicrobial for the treatment of conditions such as tinea pedis and acne. OBJECTIVES: To investigate the anti-inflammatory properties of tea tree oil on histamine-induced weal and flare. METHODS: Twenty-seven volunteers were injected intradermally in each forearm (study and control assigned on an alternating basis) with histamine diphosphate (5 microg in 50 microL). Flare and weal diameters and double skin thickness were measured every 10 min for 1 h to calculate flare area and weal volume. At 20 min, 25 microL of 100% tea tree oil was applied topically to the study forearm of 21 volunteers. For six volunteers, 25 microL paraffin oil was applied instead of tea tree oil. RESULTS: Application of liquid paraffin had no significant effect on histamine-induced weal and flare. There was also no difference in mean flare area between control arms and those on which tea tree oil was applied. However, mean weal volume significantly decreased after tea tree oil application (10 min after tea tree oil application, P = 0.0004, Mann-Whitney U-test). CONCLUSIONS: This is the first study to show experimentally that tea tree oil can reduce histamine-induced skin inflammation.

Adult↗