A 22 year old woman was incidentally found to have a hepatic small haemangioma-like mass, measuring 1.4 cm in diameter, by an ultrasonographic examination. The mass demonstrated no change in size or appearance for 6 months until the patient began to take oral contraceptives. Eventually, the mass increased to 2.0 cm in diameter after using oral contraceptives for 6 months. A histological examination suggested the mass to be typical focal nodular hyperplasia, and not hepatic adenoma. There was no further change in either size or appearance in the ensuing 1 year after the discontinuation of oral contraceptives.

Adult↗

PubMedDate not supplied

Rise of plasma myeloperoxidase during interferon therapy.

The plasma myeloperoxidase (MPO) level was evaluated using a specific radio-immunoassay (RIA) for MPO in alpha 2b-interferon (IFN)-treated patients with chronic viral hepatitis. The plasma MPO was checked before and after the initial 2 weeks use of IFN at a dose of 6 x 10(6) U/day. The mean concentration of plasma MPO was found to be markedly higher after IFN therapy than that before the therapy (421.7 +/- 34.3 vs 242.9 +/- 23.0 ng/mL, P < 0.001). The plasma MPO negatively correlated with the granulocyte count (r = -0.37, P < 0.02) and the platelet count (r = 0.49, P < 0.01, while it positively correlated with serum alkaline phosphatase (ALP; r = 0.41, P < 0.03). The plasma MPO also showed a strong correlation with plasma polymorphonuclear granulocyte elastase (PMN elastase; r = 0.73, P < 0.001). Our study thus suggests that the increased release of MPO from destroyed granulocytes is responsible for the high concentrations of the plasma MPO in patients during IFN therapy, because the plasma MPO, PMN elastase and ALP abundant in granulocytes all increased in spite of a decrease in the granulocyte count. Granulocytopenia during IFN therapy may therefore be due to the increased destruction of granulocytes in addition to a direct suppression of the bone marrow by IFN.

Agranulocytosis↗

PubMedDate not supplied

Thyrotropin releasing hormone (TRH)-induced release of 7B2 (neuroendocrine polypeptide) in vivo and in vitro using adenoma cells of a patient with acromegaly.

We demonstrated TRH-induced release of 7B2 (a neuroendocrine polypeptide) in vivo and in vitro (somatotroph adenoma cells) in a patient with acromegaly. The mean basal plasma 7B2 and growth hormone (GH) levels before operation were 142.8 +/- 3.2 ng/l and 52.4 +/- 1.6 micrograms/l (mean +/- SEM), respectively and these levels significantly rose after an i.v. administration of 500 micrograms of thyrotropin releasing hormone (TRH). After the transsphenoidal adenomectomy, the basal level of plasma GH was restored to the normal level and that of plasma 7B2 was slightly decreased. In addition, TRH-induced response of plasma 7B2 and GH disappeared post-operatively. In a primary culture of somatotroph adenoma cells obtained at surgery, TRH significantly induced secretions of both 7B2 and GH. Immunohistochemical studies showed the positive 7B2 and GH immunoreactivities in somatotroph adenoma cells. These findings strongly suggest that the somatotroph adenoma cells in this case produced and released 7B2 concomitant with GH.

Acromegaly↗

PubMedDate not supplied

Retrograde ejaculation accompanying hyperprolactinemia.

A patient, 38 years of age with 10 years of infertility, suffered from an ejaculation disorder for 2 years. Based on a diagnosis of retrograde ejaculation, sperm retained in the bladder was collected and homologous artificial insemination was carried out, but pregnancy was not achieved. The subject was examined at the Department of Urology and hormone tests were conducted showing hyperprolactinemia, hypogonadotropinemia, and hypotestosteronemia. A CT scan showed a pituitary tumor. Administration of 7.5 mg/day of bromocriptine was initiated to treat this tumor. Antegrade ejaculation recurred 1 month after administration, and pregnancy was achieved from normal sexual intercourse 5 months after administration.

Adult↗

PubMedDate not supplied

Enhanced anti-metastatic efficacy of IL-2 activated NK (A-NK) cells with novel benzothiazoles.

We have previously shown that A-NK cells when locoregionally administered accumulate within established cancer metastases and establish direct contact with both tumor cells and microvascular endothelial cells. Nevertheless, the accumulation of adoptively transferred A-NK cells into established cancer metastases is not sufficient for therapeutic efficacy in the B16 melanoma model. We have therefore attempted to enhance the anti-metastatic therapeutic efficacy of adoptively transferred A-NK cells with standard anticancer chemotherapeutic agents. We have found that chemoimmunotherapy with A-NK cells plus cyclophosphamide to be more effective than A-NK cell adoptive immunotherapy alone. We have now built on these findings, by examining the ability of novel biologic response modifiers (low molecular weight benzothiazole compounds) to augment adoptive immunotherapy with A-NK cells. Two compounds KB-R4107 (4-methoxy-2-(4-t-butylphenyl)benzothiazole) and KB-R4250 (4-methoxy-2-(4-trifluoromethylphenyl)benzothiazole) enhanced reduction of B16 melanoma pulmonary metastases mediated by A-NK cell adoptive immunotherapy. Both compounds were administered for 5 days prior to administration of A-NK cells at 100 mg/kg p.o. All experimental groups initially contained at least 7 animals and were examined for tumor burden on day 10. With B16 melanoma cells administered on day 0 and A-NK cells administered on Day 4, KB-R4107 and KB-R4250 yielded on average a 64% and 52% reduction in metastatic burden, respectively compared to an average 17% reduction using A-NK cells alone. In contrast these compounds did not diminish metastatic burden when administered alone. KB-R4107 and KB-R4250 are therefore low molecular weight, heterocyclic, biological response modifiers which can augment the anti-metastatic therapeutic effect of adoptively transferred A-NK cells.

Animals↗

PubMedDate not supplied

Mutant p53 mediated induction of cell cycle arrest and apoptosis at G1 phase by 9-hydroxyellipticine.

Wild-type p53 causes cell-cycle arrest at late G1 phase and induction of apoptosis by up- regulation of waf1 and bax, respectively. Although in many cancer cells p53 is frequently mutated and loses its functions, we have proposed that mutant p53 may be involved in the anticancer mechanism of 9-hydroxy ellipticine (9HE). It was shown using flow cytometry that 9HE (10 microM) caused induction of apoptosis in G1 phase of the cell cycle in mutant p53 (p53ala143, p53his175, orp53his273) transfected Saos-2 cells, but not in p53-deficient parental Saos-2 cells. Similar induction of apoptosis was observed 24-48 h after treatment with 9HE in mutant p53-containing SW480, SK-BR-3 and MKN-1, but not in p53-deficient KATO m cells. Using G1 phase cells isolated by centrifugal elutriation, it was confirmed that 9HE caused cell cycle arrest at G1 phase and subsequently induced G1 phase-restricted apoptosis. In accordance with the induction of arrest and apoptosis in G1 phase, 9HE caused up-regulation of waf1 and bax mRNA in mutant p53-containing cells, but not in p53-deficient parental Saos-2 cells. In control experiments, adriamycin (ADR) showed neither G1-restricted apoptosis nor elevation of bax mRNA. It is suggested that 9HE may cause G1 arrest and induction of G1 phase-restricted apoptosis by restoring the wild-type function of mutant p53 protein.

Apoptosis↗

PubMedDate not supplied

Serum leptin levels in patients with nonalcoholic chronic liver disease.

BACKGROUND/AIMS: The elevated serum leptin level of patients with alcoholic cirrhosis has been reported, however, the precise mechanism is still unknown. Leptin expression and protein synthesis have also been detected in activated hepatic stellate cells in cell cultures, which play a major role in hepatic fibrosis. We evaluated the serum leptin levels of patients with nonalcoholic liver diseases including cirrhosis and chronic hepatitis. We also investigated the hepatic clearance of leptin by determining the serum leptin level in blood samples obtained from the portal and hepatic veins. METHODOLOGY: The serum leptin level of 44 patients with nonalcoholic chronic liver disease (male/female = 21/23, cirrhosis/chronic hepatitis = 30/14) and 40 control subjects (male/female = 20/20) was determined in blood samples obtained from the antecubital vein by enzyme-linked immunosorbent assay. We also assessed the relationship between the leptin level and various biochemical tests of liver function. Additionally, we determined the leptin levels in the portal and the hepatic venous blood (nonalcoholic cirrhosis = 10, nonhepatic disease = 4). RESULTS: There were positive correlations between the serum leptin level and body mass index among males and among females in the liver disease group and in the control group. However, the serum leptin level of the liver disease group and control group did not differ significantly. Among the 44 liver disease patients, only the serum cholesterol level was significantly correlated with the serum leptin level after adjusting for sex and body mass index by multiple regression analysis. Furthermore, the leptin level in hepatic venous blood was significantly lower than that in portal venous blood. However, the ratio of [leptin level in hepatic venous blood]/[leptin level in portal venous blood] in the cirrhosis group, and that in the nonhepatic disease group, did not significantly differ. CONCLUSIONS: The serum leptin level of patients with nonalcoholic liver diseases is not elevated. On the other hand, the serum leptin level of patients with alcoholic cirrhosis has been reported to be elevated. The difference in the serum leptin level of patients with nonalcoholic liver disease and that of patients with alcoholic cirrhosis may be due to a difference in factors such as the levels of cytokines or sex steroids, and/or nutrition. Furthermore, it is likely that leptin is cleared in part by the portosystemic circulation through the liver.

Adult↗

PubMedDate not supplied

9-Hydroxyellipticine alters the conformation and DNA binding characteristics of mutated p53 protein.

The tumor suppressor protein p53 is a phosphoprotein which shows growth and transformation suppression functions. Mutational loss of p53 function is the most frequently detected genetic event in human cancers. We examined whether 9-hydroxyellipticine (9HE), a cytotoxic agent, affected the tertiary structure of mutant p53 and DNA binding characteristics. Although several types of p53 mutants were resistant to degradation by calpain, the p53 mutants treated with 9HE were markedly sensitive to calpain as well as wild-type p53. Furthermore, mutant p53 proteins isolated from 9HE-treated cells regained the ability to bind a wild-type-specific p53 DNA consensus sequence. Wild-type p53 proteins prepared from both untreated and 9HE-treated cells bound the p53 consensus sequence and were degradaded by calpain equally well. These results suggest that 9HE affects the tertiary structure of mutated p53, which results in the restoration of DNA binding characteristics.

Animals↗

PubMedDate not supplied

Hopantenic acid beta-glucoside as a new urinary metabolite of calcium hopantenate in dogs.

The metabolism of calcium hopantenate (HOPA) was studied in beagle dogs. After oral administration of 14C-labeled HOPA, 25.5% of the administered radioactivity was excreted in the urine within 24 hr, mostly in the form of unchanged drug. The only metabolite, accounting for 4.2% of the radioactivity in the urine, was isolated by HPLC. The metabolite was hydrolyzed by the treatment of beta-glucuronidase (Helix pomatia), acid phosphatase, or beta-glucosidase. These enzyme activities were not inhibited by treatment with D-glucaric acid 1,4-lactone or PO4(3-), but with D-gluconic acid 1,5-lactone, demonstrating that the metabolite is a glucose conjugate. The compound was identified as HOPA-glucoside, 4'-O-(beta-D-glucopyranosyl)-D-hopantenic acid, by GC/MS analyses after derivatization of the metabolite and the synthetic compound. This is the first reported instance of glucose conjugation to a non-acidic hydroxyl group in the metabolism of xenobiotics in mammals.

Animals↗

PubMedDate not supplied

Effect of dose of cyclosporine or FK506 and antithrombotic agents on cardiac allograft vascular disease in heterotopically transplanted hearts in rats.

BACKGROUND: We compared the severity of cardiac allograft vascular disease in rats treated with cyclosporine or FK506 and studied the effect of antithrombotic agents on cardiac allograft vascular disease. METHODS: One group each was treated with 2 and 5 mg/kg/day of cyclosporine. Two other groups were injected with heparin and dipyridamole, respectively, in addition to cyclosporine. Four other groups were similarly divided by dose of FK506 (0.1 or 0.25 mg/kg/day) and concomitant anticoagulant treatment. RESULTS: Grade of rejection and percentage stenosis of coronary arteries were lower in groups with high doses of immunosuppressive agents or with heparin. Major histocompatibility class II antigens were expressed by the endothelium of grafted hearts, and IgM and C3 were deposited in the intimal and medial layers in all groups except those administered the higher doses of immunosuppressive drugs. However, no remarkable differences in density of major histocompatibility class II antigens were found between groups demonstrating expression of these antigens. On the other hand, the intensity of IgM or C3 expression grew significantly as coronary stenosis increased in severity. CONCLUSIONS: A significant difference in severity of cardiac allograft vascular disease was not found between the groups treated with cyclosporine and FK506, and cardiac allograft vascular disease was almost entirely suppressed when doses of cyclosporine and FK506 sufficient to suppress graft rejection were administered. Our findings also showed that concomitant heparin administration reduced the extent of allograft rejection and the incidence of cardiac allograft vascular disease.

Animals↗

PubMedDate not supplied

Interleukin-2 (IL-2) activated natural killer (A-NK) cells: binding to microvascular endothelial cells and BRM enhancement of cytolytic activity.

A syngeneic in vitro system was developed for investigation of the binding of purified murine interleukin-2 (IL-2) activated natural killer (A-NK) cells to murine microvascular endothelial cells. This system is potentially useful as a model to investigate biochemical and molecular events underlying the binding of A-NK cells to microvascular endothelial cells within metastases in vivo that we have previously noted using electron microscopy. A-NK/endothelial cell binding in the syngeneic system was compared with the binding of allogeneic A-NK cells to the endothelial cells. Among test agents used to pretreat the A-NK cells for modulation of binding, lipopolysaccharide (LPS) most consistently enhanced binding. Additionally, test agents were also used in concurrent assays of A-NK cytolytic activity versus YAC-1 and P815 tumor cell targets. Polyinosinic: polycytidylic acid (poly IC), and gamma interferon (gamma IFN) were among the test agents which enhanced A-NK cell cytolytic activity.

Animals↗

PubMedDate not supplied

Augmentation of IL-2 activated natural killer cell adoptive immunotherapy with cyclophosphamide.

UNLABELLED: We have previously documented that adoptively transferred, IL-2 activated natural killer (A-NK) cells can accumulate within established pulmonary metastases. Since we have observed that increases in the accumulation of A-NK cells do not always lead to increases in therapeutic efficacy, we examined the ability of cyclophosphamide to enhance the therapeutic efficacy of A-NK cells. Animals with established B16 melanoma or Lewis lung carcinoma pulmonary metastases were treated with A-NK cell adoptive immunotherapy, either alone or following treatment with chemotherapeutic doses of cyclophosphamide. Adoptive immunotherapy studies with A-NK cells yielded at most a 30% reduction in the number of pulmonary metastases; however, cyclophosphamide (300 mg/kg) consistently reduced the size of metastatic colonies. In contrast, the combination therapy of A-NK cells plus cyclophosphamide was more effective than adoptive immunotherapy alone. In addition, polyethylene glycol IL-2 is superior to IL-2 in these studies. CONCLUSIONS: Our studies suggest that chemoimmunotherapy with A-NK cells plus cyclophosphamide may be more effective than adoptive immunotherapy alone since it results in the reduction in both the size and number of pulmonary metastases.

Adoptive Transfer↗

PubMedDate not supplied

Flavone acetic acid enhances accumulation of IL-2 activated NK cells within established metastases.

Flavone acetic acid, an agent which has been implicated in both tumor vasculature collapse and NK cell activations, has been tested recently as a potential anti-cancer chemotherapeutic agent. We have tested this agent in combination with adoptive immunotherapy using IL-2 activated natural killer (A-NK) cells in a metastatic B16 melanoma model in C57BL/6 mice. By using rhodamine-labeled A-NK cells we have been able to quantitate both the number of A-NK cells that localize within each tumor section and the percentage of the tumor area occupied by A-NK cells. This has been accomplished using an image analysis system. Flavone acetic acid (200 mg/kg, i.p.) given one day prior to the injection of A-NK cells increased the area of the tumor occupied by A-NK cells and the area of individual A-NK cells approximately 2-fold; however, it did not appear to increase the number of A-NK cells per tumor cross-section. Nevertheless, this increase did not lead to any significant change in the therapeutic efficacy of A-NK cell adoptive immunotherapy. Our studies therefore suggest that mere enhancement of A-NK cell recruitment into tumor metastases does not necessarily translate into enhanced metastatic therapeutic efficacy. Moreover, this method may be a useful tool for pre-screening of compounds which enhance the accumulation of adoptively transferred cells into tumor metastases prior to in vivo screening for therapeutic efficacy.

Adjuvants, Immunologic↗