Effect of the single major proteic fractions of the liver perchloric extract UK101 on the development of oral tumours in Syrian hamsters.

Squamous cell carcinoma of the oral cavity continues to be a major clinical problem, with about 100,000 new deaths each year worldwide. There is therefore a need to search for new tools to aid oral cancer treatment. We tested the inhibitory activity on chemical carcinogenesis of the three principal protein fractions of about 50, 14, and 8.5 kDa of the mixture UK101 derived from goat liver. These are composed principally of a glycoprotein rich in mannose residues, a protein with analogy to the heat shock protein family, and ubiquitin, respectively. The animal model employed was dimethylbenzanthracene-induced hamster cheek pouch carcinoma. Number of tumours per animal, tumour mass per animal, and proliferating cell nuclear antigen (PCNA) in non-tumour mucosa were quantified: the 14-kDa fraction was the most active; this was also confirmed by testing its corresponding recombinant material. The 50-kDa fraction was inactive, while the ubiquitin showed only low inhibitory activity. It is possible that the technique described and the results obtained could lead to an interesting clinical approach to the treatment of oral cancer.

Clinical and haematological improvement induced by etidronate in a patient with idiopathic myelofibrosis and osteosclerosis.

We report a patient with agnogenic myeloid metaplasia associated with debilitating bone pain due to increased bone turnover and osteosclerosis. Treatment with etidronate at a dose of 6 mg/kg per day on alternate months resulted in a complete recovery of bone symptoms and normalization of metabolic parameters of bone turnover; unexpectedly, a sustained haematological improvement was also observed after several months of therapy, suggesting that bone marrow microenvironment improvement was able to restore a nearly normal haemopoiesis. We suggest that diphosphonate therapy may be of value in patients with AMM and increased bone turnover.

Early and multifocal tumors in breast, salivary, harderian and epididymal tissues developed in MMTY-Neu transgenic mice.

Transgenic mice carrying various oncogenes driven by mammary gland specific enhancers develop mammary tumors usually arising in a stochastic way. The only exception is a mouse lineage (TG.NF) carrying an activated rat Neu oncogene driven by the murine mammary tumor virus long terminal repeat (MMTV-LTR) that gave rise to rapid and multifocal mammary tumors interpreted as a result of a single-step neoplastic transformation. The effect of the oncogene appeared to be specific for breast tissue, since salivary and Harderian glands as well as epididymis expressed high levels of Neu but only developed hyperplasia (Muller et al., Cell, (1988) 54, p. 105). Here we describe a transgenic mouse lineage for the MMTV-Neu, analysed up to third generation. Multifocal tumors involving mammary glands arose very rapidly in all females independently from pregnancy and in some males. Moreover, multifocal neoplasias occurred also in salivary and Harderian glands and in the epididymis at a very high rate. These data demonstrate that the Neu oncogene can induce tumors in all the tissues where it is expressed at high levels.

Acute effect of intravenous magnesium sulfate on airway obstruction of asthmatic patients.

The bronchodilating effect of magnesium sulfate (MgSO4) was studied in ten asthmatic patients with moderate to severe airway obstruction. Two grams of MgSO4 or saline in double-blind crossover design was administered IV for 20 minutes (0.40 mmol/min) and forced expiratory capacity and forced expiratory volume in one second (FEV1) were studied at intervals. Only at the end of MgSO4 infusion did FEV1 increase significantly (109% of initial values). The bronchodilating effect was short lasting and far less than that observed after salbutamol.

[ Action of low doses of vincristine, lidocaine and verapamil on DNA replication in vitro]. / Azione "in vitro" di basse dosi di Vincristina, di Lidocaina e di Verapamil sulla duplicazione del DNA.

Lidocaine and Verapamil at pharmacological doses which for single drug are not cytotoxic, when used together in vitro, inhibit DNA replication in PHA-stimulated lymphocytes but not in Jurkat cell (T-ALL line) cultures. At the same concentration the two drugs used in association with very low doses of Vincristine are cytotoxic to PHA-stimulated lymphocytes and Jurkat cells. Cytotoxic action of Doxorubicin is not increased by Lidocaine or by Verapamil or by an association of the two drugs. Changes in calcium ion concentration in the medium did not show any significant effect. These results suggest that Lidocaine and Verapamil have a common mechanism of action and have a toxic action on the same cell structure of Vincristine; furthermore the cytotoxic action of Vincristine is considerably increased. These in vitro effects could be tested in animal models.

[Verapamil inhibits the in vitro synthesis of DNA. I. Study of human lymphocyte cultures]. / Il Verapamil inibisce la sintesi del DNA "in vitro". I. Studio su colture di linfociti umani.

Verapamil in vitro reduced 3H-TdR incorporation into lymphocytes. High doses of Verapamil were lethal for cells; low doses inhibit DNA synthesis, but cell viability by the Trypan blue exclusion method is preserved. Increasing the concentration of Ca++ in the medium partially improved lymphocyte viability and 3H-TdR incorporation. Modification of the Na+ and K+ content in the medium did not interfere with Verapamil action. The inhibitory action was not completely reversible even when the cells were exposed to the drug for a short time (15'). Our results confirm the dependency of lymphocyte blast transformation on Ca++ during the first 20 h. Nifedipine did not inhibit cell replication, which suggests a different mechanism of action. Our investigation suggests that there is more than one model for calcium-antagonism and that the action of Verapamil can not be explained only by its influence on transmembrane Ca++ flux. We believe that Verapamil has a negative action on cell metabolism directly through an interaction on transmembrane Ca++ flux and on intracellular Ca++.

[Verapamil inhibits DNA synthesis in vitro. II. Study of the MCF7 and Jurkat neoplastic cell lines]. / II verapamil inibisce la sintesi del DNA "in vitro". II. Studio delle linee neoplastiche MCF7 e Jurkat.

Verapamil in cultures of mammary carcinoma MCF7 cells and Jurkat A.L.L. cells decreased the 3H-TdR incorporation. The action of Verapamil is dose-dependent. The inhibition percentage of 3H-TdR incorporation into Jurkat cells by Verapamil was lower than into MCF7 cells. No change of the mitotic inhibition was observed with higher Ca++ concentration in the medium. Modification of the Na++ and K++ content in the medium did not change action of Verapamil; this is in agreement with the independence of neoplastic cells from the culture medium. Our investigation suggests that Verapamil does not work through a block of transmembrane Ca++ flux, but rather through an intracellular interaction.

[In vitro inhibition of DNA replication by local anesthetics. Effects on human MCF7 neoplastic cells]. / Inibizione "in vitro" della duplicazione del DNA da parte di anestetici locali. Effetti su cellule neoplastiche umane MCF7.

The action of two local anesthetics (Lidocaine and Bupivacaine) on cells of mammary carcinoma MCF7 was investigated. 3H-TdR incorporation decreases in relation to the dose, and viability by Trypan blue does not significantly change but at high doses of anesthetic. Intercell adhesion decreases only at high concentration. When Lidocaine is removed after the fourth hour and Bupivacaine after the second hour the antimitotic action is irreversible. The inhibiting action of drugs is related to the cell number and unrelated to the time of adding the drug. There was no change of Lidocaine and Bupivacaine action on neoplastic cells at different concentration of Na+, K+ and Ca++ in the medium. Neoplastic cells are partially independent from Ca++ and we think the antimitotic effect of local anesthetics we observed can be due to: antagonist action to calmodulin; inhibition of aminoacylation of tRNA; inhibition of cholesterol synthesis; modification of membrane permeability which is however significant only for high concentration of the drug.

Different concentrations of local anaesthetics have different modes of action on human lymphocytes.

Lidocaine and Bupivacaine inhibit in vitro 3H-TdR incorporation into peripheral lymphocytes, in a dose dependent ratio, either under PHA stimulus or not. Lidocaine added to cultures at various times from PHA stimulus show a reduced inhibition only when added after the 24th hour. This suggests a sensitizing action of PHA. Lidocaine effect on mitosis of doses between 2000 and 500 micrograms/ml was not completely reversible even when the drug was removed after only 15 min; between 200 and 50 micrograms/ml this effect is reversible. Lymphocyte viability by Trypan Blue exclusion is clearly unrelated to mitotic inhibition; it is however dose related. High Lidocaine and Bupivacaine concentration alter adhesion of cells to plastics and decrease lymphocyte aggregation by PHA. Electrophoretic mobility of lymphocytes incubated with 2000 micrograms/ml of Lidocaine is slower compared to controls; there is no change at 200 micrograms/ml. Local anaesthetics modify steric membrane structure reducing surface charge density. Increased Ca++ in the medium containing local anaesthetic increases lymphocyte stimulation index slightly only for Lidocaine and Bupivacaine doses, which neither alter membrane function nor interfere with lymphocyte viability nor electrophoretic mobility. Increased Na+ and K+ in the medium do not affect local anaesthetic action.

[Parameters of cellular immunity in subjects with bronchial squamous cell tumors]. / Alcuni parametri dell'immunità cellulare in soggetti con neoplasia bronchiale a cellule squamouse.

We studied some cell mediated immunologic assay in a normal population and in patients with squamous cell carcinoma of the bronchus. In cancer patients lymphocyte mitogen stimulation with PHA and PWM did not show any significant difference from normal population; PPD antigenic stimulation was lower in eight patients of ten. E-total rosette forming cells count was reduced, there was no difference in E-active rosette forming cells. Electrophoretic mobility of lymphocytes showed that a part of lymphocytes have a lower electrophoretic cell migration. Lymphocytes subpopulations isolated by E rosetting technique are different in cancer patients from subpopulations separated by electrophoresis; in controls similar subpopulations were obtained by the two methods. In cancer patients cell mediated immunity shows altered functions which is though not uniformly modified. We think that serum factors and cells with blocking action cause these effects rather than intrinsic changes of lymphocytes.

[In vitro lymphocyte immunoreactivity in sarcoidosis patients. Stimulation with Kveim antigens]. / Immunoreattività linfocitaria "in vitro" in pazienti affetti da sarcoidosi. Stimolazione con antigene di Kveim.

12 patients suffering from sarcoidosis have been studied in addition to clinical and bioptic investigations, they were submitted to intradermal reaction with PPD, Candida Albicans and Kveim's antigen. Lymphocytes from the 12 patients were placed in culture and stimulated with phytohaemagglutinin (PHA), pokeweed mitogen (PWM), concanavalin A (CON-A), PPD, Candidin and Kveim's antigen. Six patients proved positive to Kveim's antigen in vivo, and four of them (66%) in vitro also. No patient was immunodepressed. Of the 6 patients negative to Kveim's antigen both in vivo and in vitro, 4 were clearly immunodepressed, as shown by the reduced mitogen response, the negative response to antigen stimulation, and to the intradermal reaction. The results are discussed in the light of recent findings regarding the role of the immunocompetent system in sarcoidosis.

[The action of insulin on human lymphocytes stimulated with mitogens "in vitro". I. Study on normal subjects]. / L'azione dell'insulina sui linfociti umani stimolati con mitogeni "in vitro". I--Studio su soggetti normali.

Insulin at 25, 50, 250, 500 and 1000 mcU/ml concentration added to lymphocyte cultures under PHA stimulation increases [3H] Thymidine incorporation. Maximum effect is obtained at 50 mcg/ml of PHA and 250 mcU/ml of insulin after 96 hours of incubation. Insulin stimulates cultures when it is added between the 12th and the 24th hour. Cultures stimulated wih high dose (250 mcg/ml) of PHA do not show any increase in mitosis. Ours and other Authors results seem to show that the mechanism of action of insulin works through specific receptors on lymphocytes. These receptors appear some hours after mitogen stimulation; our results showing maximum effect when insulin is added after 12-24 hours support this hypothesis.

[Lymphocyte immunoreactivity in subjects vaccinated with Calmette-Guerin bacillus (BCG)]. / Immunoreattività linfocitaria in soggetti vaccinati con bacillo di Calmette e Guerin (BCG).

PHA, Pokeweed and PPD lymphocytes transformation in vitro was studied in four groups of people. One group was negative to PPD skin testing; one was positive for a "naturally" acquired infection; the third group were people vaccinated with BCG in the last year; fourth group people vaccinated with BCG within one and five years. Last two groups had generally low or negative reaction to PPD skin test. PPD was used at different concentration, from 0.025 to 35 microgram/ml; maximum stimulation was at 1 microgram/ml. Stimulation index was lower in vaccinated people than in "natural" positive. Lymphocytes of people vaccinated from more than one year showed no difference from PPD negative people. Highly significant difference was observed between these two groups and people "naturally" positive and within one year of vaccination. We discuss our data and interpret the findings as showing that there are less PPD sensitive lymphocytes in patients vaccinated with BCG than in people with a "natural" infection and that this circulating population of lymphocytes disappears or decreases in the years following vaccination.

[Active E rosette test in the study of PPD hypersensitivity]. / Il test delle rosette E attive nello studio della ipersensibilità alla PPF.

In our study we show that pre-incubation of lymphocytes with PPD "in vitro", produces a significant increase in the number of E active rosettes. This increase is only for subjects with cutaneous hypersensitivity to tuberculin. We think this is a specific test and we studied the confidence interval for our results. We discuss the possibility to evaluate the immune-status of a subject, testing his lymphocytes with a panel of antigens. The active E rosette test is simpler, cheaper and easier to perform than other tests to evaluate the cellular immune-status of patients.

[Intestinal mucosa absorption and esterification of 14C oleic acid during treatment with neomycin]. / Assorbimento ed esterificazione di acido oleico C14 nella mucosa intestinale del ratto durante trattamento con neomicina.

Neomycin administered to rats subjected to lymphatic fistula and treated with 14C-labelled oleic acid did not decrease either the absorption or the esterification of this organic acid. The results appear to indicate that neomycin has no effect on the intestinal mucosa.

[14C-labelled triolein absorption and esterification in rats treated with neomycin]. / Assorbimento ed esterificazione di trioleina C14 in ratti trattati con neomicina mediante fistola linfatica.

Intragastrically administered 14C-triolein was observed in fistula-derived lymph from rats treated with neomycin and controls. Absorption was much reduced in the treated animals, indicating (contrary to the literature data) that neomycin malabsorption also occurs in this species. The slight amount absorbed followed normal routes. It is suggested that the antibiotic acts on the intestinal content and not on the mucosa.