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The cell-mediated immune response, as demonstrated by cutaneous delayed hypersensitivity, has been reported to correlate well with the clinical course in a variety of neoplastic diseases. To facilitate evaluation of cellular immunity in patients with gynecologic cancer, an immunologic index has been devised which incorporates the cutaneous reaction to standard antigens, 1-nitro,2,4-difluorobenzene, and croton oil. Thirty-six patients with invasive cervical cancer, 19 patients with cervical dysplasia or carcinoma in situ, and 77 patients with dermatologic disorders were tested. Results reported here indicate a highly significant relationship between host cellular immunity and the clinical course of the patient with a gynecologic neoplasm.

Male and female mice were given injections of ethyl carbamate at intervals after one-third or two-thirds hepatectomy. The incidence of hepatocellular tumors was increased after partial hepatectomy and correlated with both the number of cells synthesizing DNA at injection and the number of cells in mitosis. Partial hepatectomy did not affect the yields of lung tumors, liver hemangiomas, or the number and character of skin tumors if ethyl carbamate was followed by a promoting treatment with croton oil. More hepatocellular tumors occurred in males, but no sex differences in the numbers of liver hemangiomas, lung adenomas, or skin tumors were observed. The binding of an ethyl carbamate metabolite to liver DNA was fourteen times greater in males but the level of binding to lung or epidermal DNA showed no sex differences. Partial hepatectomy increased the binding to liver DNA in both sexes but did not alter the amount bound to lung or epidermal DNA. The rate of metabolism of ethyl carbamate was slower after partial hepatectomy, but this did not create a significant increase in the effective dose.

The extent of in vitro binding of 7,12-dimethylbenz(a)anthracene (DMBA) to replicating and non-replicating DNA of mouse skin epidermis was studied. Mice which were pretreated topically with croton oil in order to stimulate DNA synthesis were treated in the same area of the back with DMBA at zero time. In addition, 5-bromodeoxyuridine (BUdR) and 5-fluorouracil (5-FU) were injected at zero time and subsequently every half hour for 7.5 h. At 8 h the mice were killed and epidermal DNA was subjected to an isopycnic cesium chloride density gradient. Binding was found to both replicating and non-replicating DNA but was reproducibly greater to non-replicating DNA. BUdR substitution into replicating DNA was shown not to be a cause of reduced binding of DMBA.

A 3-stage sequential screening program for anti-inflammatory compounds is presented. It measures the 5-hr effusive response to the intrapleural injection of 5 ml of 0.075% Evans Blue-0.5% carrageenan type 7. For each stage, 2 animals are used per compound. The compounds are administered po or im to fed rats 1 hr prior to the injection of the pleural irritant. Anti-inflammatory activity is indicated by a decrease in the volume of pleural fluid. This model detects gold-, acetylsalicylic acid-, and steroid-type anti-inflammatory compounds. False negatives include: benzydamine, l -histidine, dl -pencillamine, antimalarials and immunosuppressants. Of 99 agents with diverse pharmacologic actions tested, 13 were judged “interesting” (false positives). These included ephedrine, phenylephrine, pseudoephedrine, nylidrin, hydralazine, acetaminophen, aminopyrine, desipramine, nortriptyline, bisacodyl, chlormerodrin, croton oil and bichloride of mercury.

During allergic and irritant contact dermatitis in guinea pigs produced by application of 1-chloro-2,4-dinitrobenzene and croton oils, respectively, the white cells of the peripheral blood were counted and examined for incorporation of 3H-thymidine. White cell counts remained at relatively constant levels during the dermatitis. During allergic dermatitis, however, 3H incorporation increased two- to five-fold, whereas during irritant dermatitis no increase occurred.

Fluocinonide (fluocinolone acetonide acetate) 0.05 % in FAPG base2 is a new fluorinated topical corticosteroid incorporated into a specially formulated non-aqueous synthetic base for use in psoriasis, eczema and other inflammatory dermatoses. In double-blind paired comparisons with other fluorinated topical corticosteroid creams, fluocinonide in FAPG base has generally been at least as effective as normally used concentrations of fluocortolone, flumethasone pivalate and betamethasone 17-valerate in eczema and more effective than these topical corticosteroids in psoriasis except when used with occlusive dressings, when no difference could be found.

In 100 patients with localized hand eczema and 20 patients with hand eczema and “generalization” the skin reactivity to primary irritants was compared to that of controls, thoroughly matched with respect to different variables, which may influence the basic skin reactivity. It was demonstrated that even an eczema localized exclusively to the hands provoked increased skin reactions in a remote skin area (thighs). This occurred only to some irritants (sodium lauryl sulphate, hydrochloric acid, benzalkonium chloride) but not to others. Patients with eczematous lesions also outside the hands unveiled an even more increased reactivity to the same substances but also to some other substances (sapo kalinus, croton oil and to a lesser degree, mercury bichloride and trichloracetic acid). No increased reactivity was found to cantharidin, phenol and potassium hydroxide.

Formulated corticosteroids were assayed on human skin using as a measure of anti-inflammatory activity the suppression of blisters produced by kerosene and pustules provoked by croton oil. Vascoconstriction activity was evaluated at the same time. The rank order of anti-inflammatory efficacy corresponded reasonably well with clinical judgements of comparative effectiveness although the ranking was not always the same for the identical steroid in the two models. With some steroids ointments were superior to creams while the reverse was true for other. Vasoconstriction generally correlated with anti-inflammatory activity but strong exceptions occurred.

A procedure for the fractionation and isolation of an acute inflammatory phase protein (α2-glycoprotein) and normal α1-macroglobulin from croton oil-inflamed rat serum is described. Serum from croton oil-treated rats is diluted with 0.15 M saline and treated with 1.75 M ammonium sulphate. The resultant precipitate is redissolved and subjected to gel filtration on a column of Sephadex G-200. The first peak eluted from the column, consisting primarily of α2-glycoprotein and α1-macroglobulin, is then subjected to preparative starch block electrophoresis to separate the two macroglobulins. Electrophoretic and immunochemical analyses showed that the α2-glycoprotein preparation obtained by this method was homogeneous and free from contaminating serum proteins.

1. Sodium soaps prepared from olive oil, croton oil, cod-liver oil, linseed oil, etc., have the property of inhibiting the action of trypsin and leucoprotease. 2. The activity of these soaps is dependent upon the degree of unsaturation of the fatty acids and is in proportion to their iodin value. 3. Saturation of the acids with a halogen (iodin) causes a loss of this property. 4. Soaps of the saturated fatty acids tested do not have this influence on ferments.

1) In guinea pig, the lip was irritated with repeated injection of croton oil for various length, of time, and the changes produced in the lip, superior cervical and caeliac sympathetic ganglia, and various organs were investigated histopathologically, 2) In the first group of animals to which croton oil was injected for a short period of time, . only edema of the lip was observed, whereas in the second group to which injection was repeated for longer period, necrosis and/or cicatrix formation were remarkable.3) The sympathetic ganglia showed little change in the animals of the first group, while in the, second group they showed degenerative changes mainly consisting of atrophy and swelling of the cells, . vacuolization or destruction of the cytoplasma, chromatolysis, swelling, atrophy, deformation and dest-ruction of nuclei. The longer the lip was exposed to the irritation the higher was the degree of the changes. Changes in the superior cervical and caeliac sympathetic ganglia were similar to each. other.4) In various organs changes due to circulatory disturbance such as congestion and hemorrhage as-well as various degenerative changes were observed. These changes were more pronounced in group I..5) On the basis of these findings, the author explained the shock symptoms produced by irri-tation of the lip as a manifestation of Reilly's phenomena, and offered a postulation that the irritation to the lip was transmitted to the autonomic nervous system of the entire body through the superior cervical ganglion working as one of the mediators of the stimulus transmission.6) Further more, the cause of novocain shock was discussed clinically from the viewpoint of Reilly's theory.

Various acute phase reactants (APR) have been studied in rats with different types of inflammation. These APR (serum albumin, serum globulins, plasma fibrinogens, serum alpha-2-glycoprotein [GP], total serum glycoproteins, serum mucoproteins) and neutrophils and lymphocytes reflect the onset, intensity, and duration of many types of experimental inflammation. The APR and cellular changes, like the inflammatory processes eliciting them, are stereotyped. Chemically induced, chronic immunologic, and delayed hypersensitivity-type acute inflammatory reactions cannot be distinguished by the APR. The most sensitive and quantitatively reliable index for the assessment of the “anti-inflammatory” action of the various steroidal and nonsteroidal drugs is the behaviour of the serum alpha-2-glycoproteins. This is also the most sensitive index of the “anti-inflammatory” action of adrenocortical steroids (or their absence, as shown in adrenalectomized rats). The various serum protein changes in rats with inflammation occur in response to greater sequestration rates at the sites of inflammation. Acute and chronic inflammation can be induced with a variety of substances affecting cell membranes (Filipin, digitonin, saponin, nystatin, d -α-tocopherol, vitamin A, and etruscomycin, to name but a few). These inflammatory reactions are inhibited in vivo by both steroidal and nonsteroidal “anti-inflammatory” drugs. We have visualized the process of inflammation as a wave of leaky membranes. The protective effects of nonsteroidal “anti-inflammatory” drugs on heat-induced and hypotonic erthrocyte lysis in vitro partially confirm the idea. The anti-arthritic effects of cytarabine (cytosine arabinoside, an immunosuppressant drug) also have been studied. Cytarabine has no systemic or local “anti-inflammatory” effect in a variety of experimental assay systems. It inhibits completely the onset of adjuvant arthritis and renders animals tolerant to further adjuvant inoculations. The APR cannot be used exclusively to distinguish between the different steroidal, non-steroidal, and immunosuppressant types of “anti-inflammatory” drugs. They are useful, however, in the overall assessment of the “anti-inflammatory” effects of drugs in all types of inflammation. The APR should be incorporated as standard procedures in laboratories “discovering” new “anti-inflammatory” drugs. Emphasis is placed on the APR as necessary requirements for the assessment of “anti-inflammatory” drugs because for example, Filipin-induced hind-paw edema can be inhibited in vivo by the i.p. administration of digitonin, another lytic agent which produces inflammation.

(1973). The Function and Mechanism of Promoters of Carcinogenesis. CRC Critical Reviews in Toxicology: Vol. 2, No. 4, pp. 419-444.

Epidermal tumors were induced in Swiss female mice by a topical application of 9,10-dimethyl-1,2-benzanthracene solution followed by repeated applications of croton oil solutions. Fourteen benign and malignant tumors were sampled 25 weeks after the treatment had begun, fixed in osmium tetroxide or potassium permanganate, and embedded in Epon. Sections stained with lead hydroxide were examined. In three tumors defects of the epidermal basement membrane were seen. These defects accompanied local invasion of the tumors. The possible mechanisms of the development of this unusual anatomical situation are discussed.

Summary.— The 3H-Thymidine (3HT) nuclear labelling index of “round” cells of the dermal infiltrate in allergic and irritant patch test reactions in man has been compared. The skin of 15 positive allergic patch test reactions (to different allergens) and of 15 positive irritant reactions (5 to croton oil, 5 to potassium hydroxide and 5 to white spirit) was biopsied 48 hours after the application of the chemical. Thin slices of each specimen were incubated for 60 min in the presence of 3HT. then fixed and prepared for light microscopy radioautography. Two thousand “round” cells of the infiltrate were counted in each specimen. The mean values of the 3HT labelling indices (±S.E.) were 2·91±0·18%, in the allergic group and 0·58 ± 0·06% in the irritant group. The difference between the 2 means was highly significant (P 0.05). All the labelled cells were mononucleated. This method could be useful to differentiate clinically dubious allergic and irritant reactions.

Newborn ICR Swiss mice received a single maximum tolerated subcutaneous injection of the test chemicals. Six months later the lungs were examined and the mean number of adenomas and percentage of animals with adenomas were determined. A significant increase in mean number of adenomas was observed after urethan (1000 or 1500 mg/kg) and N -nitrosodiethylamine (50 mg/kg) treatment. Liver tumors were also observed after the N -nitrosodiethylamine treatment. Mean adenoma counts comparable to control values were observed after treatment with thiourea (2500 mg/kg), mesidine (250 mg/kg), aflatoxin (2.5 mg/kg), 2,2-bis( p -chlorophenyl)-1,1,1-trichloroethane (DDT, 15 g/kg), croton oil (0.5 ml/kg), dodecylbenzene (12.5 ml/kg), or dimethyl sulfoxide (6 ml/kg). The vehicle control incidence of lung adenomas, after saline or peanut oil injection, varied from 2 to 14%, and the mean number of adenomas was 0.02 to 0.14. The bioassay procedure appears to be negative to promoter substances. The effect of nitrogen mustard on lung adenoma incidence was reduced when the vehicle was peanut oil or tricaprylin.

These observations tend to support Selye's hypothesis that the effect of the adrenal steroids is to inhibit the capacity of the tissues to respond to a noxious stimulus by inflammatory reaction. In these experiments, the inflammatory response was well under way before the hydrocortisone was introduced into the injured tissues; however, the arthritis caused by the turpentine and the granulomatous process by which the tale was isolated from the joint cavity continued to progress. The gross appearance showed a diminution of the vascular-dilatation element of inflammation in most instances on the treated sides and the microscopic appearance showed as a rule fewer infiltrating phagocytic cells on the treated sides. These findings suggest not only that there is an inhibition to the progress of the inflammatory response, but also that the tissues tend to return to a normal state more rapidly after treatment. In our experiments, the injury was relatively mild and the hydrocortisone seemed to exert a favorable influence on recovery, although the difference between the treated and the untreated sides was not great. If the hydrocortisone had been administered at the time of the injection of the irritant, it is probable that the effect would have been more marked. There was no tendency for the hydrocortisone to accentuate the damage and to cause necrosis, as was noted by Selye in his experiments with croton oil. It seems probable to us, however, that the thick barrier between the skin and the inside of the granuloma pouch in Selye's experiments, in which the skin did not become necrotic, was a mechanical one and that the steroid really did not actually enhance the necrotizing effect of the croton oil. In more severe injury to tissues, as Selye showed in his experiments, the anti-inflammatory effect of hydrocortisone may result in greater damage to the tissues involved, because the inflammatory reaction may have a protective effect by creating a shield. In the clinical application of these principles, it seems logical to assume that hydrocortisone would be most useful in those pathological conditions which are inherently localized and self-limited, in which the symptoms are largely due to the inflammatory response per se , and which are either progressing or in which the noxious agent is continuing to act.

The time of appearance of the tumors, and the cumulative response, varied with the dose of a fraction of croton oil which was applied on the initiated skins of mice. The present results have not established a rigorous dose-response relationship; they indicate nevertheless a definite quantitative trend.

On a preliminary test, anti-inflammatory and analgesic dose-related activities on rats were observed for the aqueous fraction of Urera baccifera; this extract was bioassay-guided fractionated and the final aqueous fraction was used according the ethnobotanical use. Carrageenan-induced edema (n=6), was used as an assay in the fractionating process. The anti-inflammatory and antinociceptive properties of the final aqueons fraction were studied using in vivo models. For the anti-inflammatory activity rat paw edema (n=6), pleurisy induced by carrageenan (n=6) and ear edema induced by topical croton oil (n=6) models were used, and tail-fIick test (n=6), abdominal constrictions induced by acetic acid (n=6), and formalin test (n=6), were used for the alltillociceptive activity. The tests perfonned showed an inhibition effeet on !eukocyte rnigration, and a reductioll on pleural exudate, as well as dose-dependant peripheral analgesic activity, al a range of 25-100 mglk:g i.p. The final aqueous fraction contains most of the anti-inflammatory activity of the plant U. baccifera. A possibJe mechanism of action is discussed and based on the results we conclude that this plant has a potential. for both anti-inflammalory and analgesic activity al the c1inical level.

USING a modification of the “granuloma-pouch” technique—which had originally been designed for the quantitative assessment of inflammation (Selye, 1953a, 1953b)—it has been possible to expose connective tissue to 100% croton oil for periods of many months and thus to induce transplantable malignant neoplasms by treatment with this inflammatory irritant alone (Selye, 1954a). One of the cancers so induced, “croton-pouch tumor No. 2,” originated as a typical fibrosarcoma in the croton-oil treated animal, but after a few animal-passages, it developed a tendency to form cartilaginous and bony islets so that it now bears the characteristics of a fibro-chondro-osteosarcoma. It takes in 100% of the cases when transplanted as a tissue-suspension, intramuscularly or into a subcutaneous air-sac (such as is used for the production of granulomapouches), into Sprague-Dawley rats, but Wistar rats appear to be immune to it. Grown in the granuloma-pouch it produces large quantities of a viscous exudate which is extremely ...

In explaining fixation of various foreign substances at the site of inflammation as the result of mechanical obstruction by thrombosed lymphatics and by a fibrin network in tissue spaces of the affected area, 3 types of evidence were presented: (1) the appearance of thrombosed lymphatics and of a fibrinous network; (2) the inability of substances to enter the site of inflammation when injected at its periphery; (3) finally, the inhibition of fixation upon the addition to the inflammatory irritant of a fibrin solvent, concentrated urea.1, 2, 3 It has been pointed out that both factors, coagulated plasma in tissue spaces and fibrinous clots occluding lymphatics at the site of inflammation, can well account for fixation. Such clots in lymphatics have been seen in vivo as well as in fixed preparations.4 At no time, however, has the writer maintained that either one of these 2 factors was more responsible for fixation than the other. In the types of inflammation studied previously with aleuronat or croton oil,...

The present investigation was concerned with the effect of the progestogen hormone, DuphastonR, on the inflammatory reaction of rats. The granuloma pouch technique elaborated by Selye was used. The injection of 25 ml of air into the dorsum of the rat created a subcutaneous sac, into which the hormone and the irritant were deposited. Injections of 1 % solution of croton oil (the irritant) into this sac produced a pronounced inflammatory reaction in the connective tissue of the wall of the sac and formed a thick granulomatous membrane. If the progestogen was deposited into the granuloma pouch at the same time as the eroton oil or on the previous day, the membrane, compared with that induced solely by the croton oil was much thinner. Progestogen itself injected into the air sac produced no granulomatous membrane at all. These results seem to indicate that the hormone progestogen may depress the inflammatory reaction of the connective tissue induced by the croton oil. In current experiments the effect of estradiol, gonadotropin and relaxin on the granuloma pouch inflammation is being studied.

The Sulphydryl Levels of Mouse Skin During Treatment with Polycyclic Hydrocarbons and Croton Oil

Phorbol ester acetate was produced by the esterification of the free hydroxyl of phorbol ester C (the tumor promoting agent from croton oil). The phorbol ester acetate was used to promote skin tumors in female mice that had previously received a subcarcinogenic dosage of 7,12 DMBA. The morphology of the tumor revealed an epidermal hyperplasia as well as a hyperkeratosis of the stratum corneum. There was a marked increase in the intercellular space in the strata basale and spinosum. These cells also demonstrated an increase in ribosomes and polysomes. Many of the mitochondria of these cells revealed intramitochondrial granules that resembled lipid droplets. Phorbol ester- 14 C acetate was prepared by acetyl-2- 14 C esterification of the free hydroxyl of the phorbol ester C. The phorbol ester- 14 C acetate was applied to papillomas that were initiated with 7,12 DMBA and promoted with phorbol ester acetate for 90 days. Grains due to the phorbol ester- 14 C acetate were observed overlying the loose chromatin of the nuclei and tonofibrils.