Treatment of experimental ovarian carcinoma with monthly injection of the agonist D-Trp-6-LH-RH: a preliminary report.

Hormones, particularly gonadotropins, have been implicated in the development of ovarian cancer. Chronic administration of agonistic analogs of luteinizing-hormone releasing-hormone (LH-RH) induces an inhibition of the pituitary-gonadal axis. The blockade of the release of luteinizing-hormone and follicle-stimulating hormone (FSH) may exert a possible therapeutic effect on ovarian cancer. We examined the results of prolonged administration of D-Trp-6-LH-RH, an agonistic analog of LH-RH in experimental ovarian cancer. We used the recently developed ovarian cancer model in rats, which is produced by treatment of pregnant rats with N-nitrosobis(2-oxopropyl)amine (BOP), following which a high incidence of ovarian tumors are induced in the offspring. In morphologic aspects the induced tumor resembles human ovarian neoplasms. Once a month administration of a delayed release preparation of microcapsules of D-Trp-6-LH-RH prolonged the survival and decreased tumor growth and the incidence of metastases. Additional experimental and clinical studies are needed to determine the efficacy of the treatment with LH-RH analogs in ovarian cancer.

Analysis of nm23-H1 expression in breast cancer. Correlation with p53 expression and clinicopathologic findings.

Metastasis is the most frequent cause of death in patients with breast cancer. The nm23-H1 and p53 genes have been involved in the development of breast cancer metastasis. We have analyzed the correlation between the expression of nm23 protein and several established clinicopathologic factors. Our results show that the antimetastatic role of nm23-H1 is not related to the cell proliferative status or tumor grade and that it is not associated with the expression of p53. We also demonstrate a strong inverse relationship between the expression of nm23-H1 protein, lymph node metastasis and vascular invasion. These data support the antimetastatic role of the nm23-H1 gene and suggest that nm23-H1 and p53 genes may be involved in different steps of the metastatic process.

Chronic gastritis associated with Helicobacter pylori. Correlation between histological and bacteriological findings.

Biopsy specimens of gastric and duodenal mucosa from 326 patients were examined bacteriologically and histologically to determine the correlation between chronic gastritis and H. pylori colonization. H. pylori was identified in 111 (66.5%) patients with evidence of chronic gastritis and in 97 (82.2%) individuals who had gastritis associated with other pathology (gastric o duodenal ulcer, carcinoma o bulboduodenitis). The spiral bacteria was found more frequently in specimens with chronic superficial gastritis (88/107) and no significant difference was observed between the grade of activity of gastritis and H. pylori colonization. Giemsa stain was the most suitable method for detecting H. pylori in histological sections. By electron microscopy the microorganism was seen on the surface of the gastric mucosa, beneath the mucous layer, and more occasionally in intercellular junctions and the gastric pit.

Experimental thioacetamide-induced cirrhosis of the liver.

Hepatic cirrhosis is a complex disease in which several biological, biochemical and chemical alterations are combined, none of these alone being sufficient for diagnosis. The morphological characteristics of the final stages of cirrhosis are well known, but the initial lesions and intermediate stages still have not been fully clarified. An experimental model of hepatic cirrhosis by chronic administration over 30 weeks of thioacetamide (50 mg/kg twice weekly) to female Wistar rats has been produced. In a macroscopic, microscopic and ultrastructural study. The different lesions that appeared were evaluated according to the dose of the toxic agent administered up, until hepatic cirrhosis was finally installed; this was after 60 doses of the toxic agent (30 weeks). Discussion is made of the different types of administration and the doses employed to obtain a suitable survival rate for these cases; in our experiments this was 95%. It has been demonstrated in both human and experimental pathology that once the disease itself has been installed, currently there is no rational or useful treatment for it. A beneficial effect has been demonstrated for certain substances, improving the initial and intermediate lesions, so we conclude by stating that it is necessary to further study the hepatic lesions preceeding cirrhosis. Knowledge of these lesions could form the basis for establishing a useful and rational therapy for such cases.

Transrectal fine needle aspiration biopsy of the prostate combining cytomorphologic, DNA ploidy status and cell cycle distribution studies.

Fine needle aspiration (FNA) cytology of the prostate is becoming a common diagnostic procedure, and DNA flow cytometry (FCM) data have been shown to correlate with the pattern of evolution of prostatic carcinoma, thus emphasizing the importance of assessing both parameters together. The aim of the present paper is to analyze the presence of DNA aneuploidy, cell cycle distribution and their relationship with the cytologic grade in transrectal fine needle aspiration prostate biopsies from 78 consecutive patients. Herein we studied the DNA ploidy status, the cell cycle distribution and their relationship with cytologic grade in transrectal FNA biopsies of the prostate from 78 consecutive patients -47 benign hyperplasias and 31 carcinomas- as analyzed by a reproducible FCM method for single cell suspension preparations, data acquisition and analysis. The presence of DNA aneuploidy was detected in 39% of the carcinomas and it was found to be a specific marker for prostatic carcinoma since all benign hyperplasia cases were diploid. Moreover, the incidence of DNA aneuploidy increased progressively from well-differentiated to moderately-differentiated and poorly-differentiated carcinomas (p = 0.005). Regarding cell cycle distribution, carcinomas displayed a higher proportion of both S-phase (p = 0.0003) and G2/M-phase (p = 0.0006) cells with respect to benign hyperplasias. Aneuploid cases also showed a greater proliferation rate as compared to the diploid carcinomas, regardless of their cytopathologic grade (p = 0.00001). Despite the fore-mentioned results, these correlations were far from being absolute, suggesting that combined assessment of these parameters should give additional information for the clinical management of prostatic disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Modifications of "A cells" in the development of experimentally induced pancreatic ductal adenocarcinoma: an ultrastructural and immunohistochemical study.

In the hamster-BOP model, modifications occur in the endocrine portion of the pancreas during the induction of well-differentiated ductal carcinomas. In the present work, using immunohistochemical techniques it was possible to observe that the A cells in preserved islets and in the initial phase of the carcinogenic process were localized in the peripheral part of the islets. In animals with tumours, A cells were found to form part of the tumour glands and/or the tumoral stroma; this localization was seen to depend on the developmental phase of the tumour. A focal dispersion of A cells was observed in the animals with ductal carcinoma.

Ultrastructural study of the initial phases of pancreatic carcinoma induced by BOP in the Syrian golden hamster.

A serial study was carried out on the lesions induced by N-nitroso-bis(2-oxopropyl)amine (BOP) in the Syrian golden hamster until the appearance of pancreatic ductal carcinomas. During the initial phase, first findings were cytolysis of acinar cells close to blood vessels and other cells, together with a loss of zymogen granules from the cytoplasm, and an increase in the diameters of the acinar lumens. After week 11 a proliferation of ductule-like cells was observed. We consider that a minimum proliferation of cells at the acinus-ductule junction would give rise to pseudoductules composed of remains of acinar cells together with ductule-like cells.

Experimental carbon tetrachloride-induced cirrhosis of the liver.

The authors produced an experimental model of cirrhosis by administering carbon tetrachloride (CCl4) intraperitoneally over 46 weeks to female Wistar rats at a dose of 0.2 ml of a solution of CCl4 in 33% mineral oil. The cytotoxic effect on the liver of this substance was evaluated in different developmental stages of the process by macroscopic, microscopic and ultrastructural studies.

Flow cytometry in the diagnosis of cancer.

Flow cytometry has rapidly expanded from basic research to clinical laboratories mainly due to its unique characteristics regarding cell analysis. Among the clinical uses of flow cytometry cancer represents one of the most relevant. Several applications of flow cytometry can currently be applied to the study of cancer, including the detection of tumour cell DNA aneuploidy, the analysis of tumour cell proliferation and the immunophenotyping of leukemias. Although standardized flow cytometry protocols for these applications are scanty, the clinical value has been clearly established. The presence of DNA aneuploidy and a high proportion of S-phase tumour cells have been associated with tumour malignancy and a poor prognosis. The immunophenotype of leukaemia is of great help both for the diagnosis and classification of chronic lymphoproliferative disorders and acute leukaemias, especially in acute lymphoblastic leukemia cases and the M0, M3-variant, M6 and M7 acute myeloblastic leukaemia subtypes. In addition, it allows the identification of relatively rare leukemia cases such as the biphenotypic and the Nk-cell lineage leukemias. The development of flow cytometry is continuously bringing new applications into the clinical laboratory in the area of cancer diagnosis.

[Colorectal cancer: retrospective analysis of 762 cases]. / Cáncer colorrectal: análisis retrospectivo de 762 casos.

A study carried out during 1976-1988 on patients with colorectal cancer revealed an increase in the incidence of this disease of 150%, with a sex ratio of 1:1.17, there being a slight predominance in males. In the patients studied, 3.41% were older than 40. In patients in the 60-70 age group, 63.4% of the cases were diagnosed. The most frequent location of the tumor is the rectum (24.4%) and the least frequent (1.57% and 1.04%, respectively) in the hepatic and splenic angles. This was observed in all the years comprising the study. Adenocarcinoma was the commonest type of tumor, there being a highly significant association (p less than 0.001) between the histological type of cancer and the state of infiltration observed at the time of diagnosis.

[Correlation among endoscopic, histological, and microbiological findings in gastroduodenal pathology associated with Helicobacter pylori]. / Correlación entre los hallazgos endoscópicos, histológicos y microbiológicos de la patología gastroduodenal asociada a Helicobacter pylori.

We studied 152 patients referred for endoscopy of the upper gastrointestinal tract. Samples of the gastric mucosa and fundus were taken and processed for microbiological and histological study using conventional methods. Acute lesions to the gastrointestinal mucosa were those most frequently observed in endoscopy (28.3%). The overall prevalence of chronic gastritis was 90.8%. The prevalence of infection by H. pylori was 59.8%. All patients in which gastric ulcer or malignant lesions were observed had some kind of chronic gastritis. Duodenal ulcer was the lesion with the highest prevalence and statistical significance with respect to the existence of H. pylori infection. There was a highly significant correlation between H. pylori gastric colonization and lesions of the diffuse antral type and with gastric activity. The significant correlations among the endoscopic, microbiological and histological findings are discussed.

[Effect of (+) cyanidanol-3 on experimental liver cirrhosis induced by carbon tetrachloride]. / Efecto del (+) cianidanol-3 sobre la cirrosis hepática experimental inducida por el tetracloruro de carbono.

Liver cirrhosis is a very common illnesses and currently its treatment remains unpromising. In a research line devoted to studying the hepatotoxicity of certain substances we have evaluated the lesions that appear prior to the occurrence of cirrhosis and hepatocarcinoma in order to test, in different assays liver-protecting substances such as cyanidanol-3 that have been seen to lead to an improvement in morphological alterations in the liver and even to reverse such lesions. The mechanisms of action of this compound are discussed. The authors believe that considerable work remains to be carried out but that some day some liver-protecting substances that yield good experimental results may be applied to clinical situations.

[Double gastric neoplasia (lymphoma/adenocarcinoma) associated with alpha-1 antitrypsin deficiency. Presentation of a case and review of the literature]. / Doble neoplasia gástrica (linfoma/adenocarcinoma) asociada a déficit de alfa-1 antitripsina. Presentación de un caso y revisión de la literatura.

A case of simultaneous double gastric primary neoplasia, lymphoma and adenocarcinoma, is presented. This association having been described in only 11 cases, our case also possessing a deficit of alpha-1-antitrypsin, the implication of which has not been clarified. We review the literature on the variety of gastric neoplasia associations, highlighting the unknown pathogenic aspects.

Histologic changes in the rat prostate cancer model after treatment with somatostatin analogs and D-Trp-6-LH-RH.

Histopathologic changes produced during the treatment of Dunning R3327 prostate cancer with new superactive somatostatin analogs (RC-121 and RC-160) and D-Trp-6 analog of luteinizing hormone-releasing hormone agonist (D-Trp-6-LH-RH) were studied. A significant reduction of the tumor weight could be observed in all treated groups, but the greatest decrease in the tumor volume was seen in the groups receiving the combination of the somatostatin analog and D-Trp-6-LH-RH. Histologically, the treatments resulted in a loss of the tumorous glandular elements and the proliferation of the stromal cells. In the tumors treated with somatostatin analogs, the amount of connective tissue was greatly increased and was accompanied by the appearance of thick collagenous fibers. In the D-Trp-6-LH-RH treated groups, regressive changes in the epithelium were seen in addition to the proliferation of connective tissue. The greatest histologic improvement was observed in the group treated with the combination of RC-160 and D-Trp-6-LH-RH. This histopathologic evaluation clearly supports our contention that superactive analogs of somatostatin greatly potentiate the inhibitory effect of D-Trp-6-LH-RH on the growth of Dunning prostate tumors and may improve the clinical response in patients with prostate cancer.

Treatment of nitrosamine-induced pancreatic tumors in hamsters with analogs of somatostatin and luteinizing hormone-releasing hormone.

Pancreatic ductal adenocarcinoma was induced in female Syrian golden hamsters by injecting N-nitrosobis(2-oxopropyl)amine (BOP) once a week at a dose of 10 mg per kg of body weight for 18 weeks. Hamsters were then treated with somatostatin analog D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Trp-NH2 (RC-160) or with [6-D-tryptophan]luteinizing hormone-releasing hormone [( D-Trp6]LH-RH) delayed delivery systems. Microcapsules of somatostatin analog RC-160, designed to release a dose of 5 micrograms/day, were injected twice a month and microcapsules of [D-Trp6]LH-RH, calculated to liberate 25 micrograms per day, once a month. After 18 weeks of BOP administration, the hamsters were divided into three groups of 10-20 animals each. Group I consisted of untreated controls, group II was injected with RC-160, and group III was injected with [D-Trp6]LH-RH. A striking decrease in tumor weight and volume was obtained in animals treated with [D-Trp6]LH-RH or with the somatostatin analog RC-160. After 45 days of treatment with either analog, the survival rate was significantly higher in groups II and III (70%), as compared with the control group (35%). The studies, done by light microscopy, high-resolution microscopy, and electron microscopy, showed a decrease in the total number of cancer cells and changes in the epithelium, connective tissue, and cellular organelles in groups II and III treated with the hypothalamic analogs as compared to controls. These results in female hamsters with induced ductal pancreatic tumors confirm and extend our findings, obtained in male animals with transplanted tumors, that [D-Trp6]LH-RH and somatostatin analogs inhibit the growth of pancreatic cancers.

Combination of a long-acting delivery system for luteinizing hormone-releasing hormone agonist with Novantrone chemotherapy: increased efficacy in the rat prostate cancer model.

The combination of hormonal treatment based on a long-acting delivery system for the agonist [6-D-tryptophan]luteinizing hormone-releasing hormone ([D-Trp6]-LH-RH) with the chemotherapeutic agent Novantrone (mitoxantrone dihydrochloride) was studied in the Dunning R3327H rat prostate cancer model. Microcapsules of [D-Trp6]-LH-RH formulated from poly(DL-lactide-co-glycolide) and calculated to release a controlled dose of 25 micrograms/day were injected intramuscularly once a month. Novantrone (0.25 mg/kg) was injected intravenously once every 3 weeks. Three separate experiments were carried out. When the therapy was started 45 days after transplantation and continued for 70 days, tumor volume in the presence of the microcapsules (966 +/- 219 mm3) or Novantrone (3606 +/- 785 mm3) given alone was significantly decreased compared to controls (14,476 +/- 3045 mm3). However, the combination of microcapsules and Novantrone caused a greater inhibition of tumor growth (189 +/- 31 mm3) than the single agents. Similar effects were seen when the percent increase in tumor volume was examined. Tumor volume increased 10,527 +/- 1803% for the control group. The inhibition of growth caused by the [D-Trp6]LH-RH microcapsules alone (672 +/- 153% increase in volume) was again greater than that caused by Novantrone alone (2722 +/- 421% increase). The combination of the two agents was again the most effective, resulting in an increase in tumor volume of only 105 +/- 29%. Control tumors weighed 30.0 +/- 6.5 g. Tumor weights were much less in the groups treated with either microcapsules (3.28 +/- 0.69 g) or Novantrone (19.53 +/- 3.3 g) alone. The lowest tumor weights after 70 days of treatment were obtained in the group that received the combination of [D-Trp6]LH-RH microcapsules and Novantrone (1.02 +/- 0.2 g). Testes and ventral prostate weights were significantly diminished by the administration of microcapsules of [D-Trp6]LH-RH alone or in combination with Novantrone. In both of these groups, luteinizing hormone and prolactin levels were reduced and serum testosterone was suppressed to undetectable levels. Similar results were obtained in two other experiments in which the duration of treatment was 60 or 105 days. These results suggest that the overall response could reflect the inhibition of proliferation of hormone-independent cancer cells by Novantrone in addition to the suppressive effect of [D-Trp6]LH-RH on the growth of androgen-dependent tumor cells. The administration of Novantrone in combination with microcapsules of [D-Trp6]LH-RH might produce a better clinical response than LH-RH analog alone in patients with advanced prostate carcinoma.

Flow cytometric DNA analysis of fine-needle aspirates of prostatic benign lesions.

Fine-needle aspiration samples from 54 consecutive patients suffering from prostatic benign lesions were analyzed for both cytological diagnosis and DNA cell content, by means of standard cytological criteria and flow cytometry, respectively. Of these patients, 13 displayed prostatitis: three being classified as acute prostatitis, 7 as chronic and 3 as granulomatosous prostatitis. All cases had a diploid DNA cell content, with a mean percentage of S-phase cells of 1.8 +/- 2.7%. Although no major differences were detected between patients with and without prostatitis, the former group had a slightly higher proportion of S-phase cells (1.4 +/- 2.2% vs 2.7 +/- 3.7%). Moreover, within those patients with prostatitis, acute prostatitis cases showed the highest proliferation rates although differences did not reach statistical significance.

Histological findings in the rat prostate cancer model during treatment with a luteinizing hormone-releasing hormone agonist and novantrone.

Morphological changes produced by the treatment with the D-tryptophan-6 analog of luteinizing hormone-releasing hormone (D-Trp-6-LH-RH) and mitoxantrone (novantrone) were studied in the Dunning R3327H rat prostate cancer model. Microcapsules of D-Trp-6-LH-RH, calculated to release a controlled dose of 25 micrograms/day, were injected intramuscularly once a month. Novantrone (0.25 mg/kg body weight) was injected intravenously once every 3 weeks. The pathology of tumors was studied in two experiments. In the first experiment, the treatment was started 135 days after tumor transplantation, and the therapy was continued for 105 days. In the second experiment, the treatment was initiated 45 days after tumor transplantation, and was carried on for 70 days. The rats were divided into four groups: 1) untreated control, 2) microcapsule-injected, 3) novantrone-injected, and 4) combination of microcapsules and novantrone-injected. In both experiments, similar results were obtained, which included significant reduction in the tumor volume and weight, a very striking decrease in the number of epithelial tumoral cells with atrophy of the glandular epithelium, and an increase in the stromal connective tissue. All these changes were more prominent in the group treated with the combination of microcapsules and novantrone than in the groups treated with microcapsules or novantrone alone. Pathological results support the view that combined therapy may be more efficacious than the treatment with single agents.

Protective effects of analogs of luteinizing hormone-releasing hormone against x-radiation-induced testicular damage in rats.

Possible protective effects of the agonist [D-Trp6]LH-RH (analog of luteinizing hormone-releasing hormone in which Gly-6 is replaced by D-tryptophan) and antagonist N-Ac-[D-Phe(pCI)1,2,D-Trp3,D-Arg6,D-Ala10]LH-RH against testicular damage caused by x-radiation were investigated in rats. Three months after being subjected to x-irradiation of the testes with 415 or 622 rads, control rats showed marked reduction in the weights of the testes and elevated levels of LH and follicle-stimulating hormone (FSH), indicating tubular damage. Histological studies demonstrated that, in testes of rats given 415 rads, most seminiferous tubules had only Sertoli cells and no germinal cells, and, in the group given 622 rads, the depression of spermatogenesis was even more marked. Rats pretreated for 50 days with LH-RH antagonist (1000 micrograms/kg of body weight per day) showed a complete recovery of testicular weights and spermatogenesis 3 months after 415 rads and showed partial recovery after 622 rads, and LH and FSH levels returned to normal in both of these groups. Thus, pretreatment of rats with LH-RH antagonist, by reversibly inhibiting gonadal function, protected the germinal cells of the testes against damaging effects of x-rays. Three experiments were also carried out in which the rats were pretreated for 1-2 months with long-acting microcapsules of the agonist [D-Trp6]LH-RH, liberating 25 micrograms of the agonist per day. Some rats were then subjected to gonadal irradiation with 415 or 622 rads and allowed a recovery period of 2-4 months. In spite of pretreatment with [D-Trp6]LH-RH, testicular weights were significantly lower and LH or FSH levels were elevated in the irradiated groups as compared with nonirradiated controls. The recovery of spermatogenesis was incomplete, and there was a decrease in the number of germinal cells after 415 rads and especially after 622 rads. On the basis of testicular weights, histology, and gonadotropin levels, it could be concluded that the agonist [D-Trp6]LH-RH did not protect the rat testes exposed to 622 rads and, at most, only partially protected against 415 rads. These results suggest that pretreatment with LH-RH antagonists and possibly agonists, might decrease the testicular damage caused by radiation and accelerate the recovery of reproductive functions.