Essential role for Ras signaling in glioblastoma maintenance

Malignant gliomas can be induced in mice through the combined expression of activated forms of both KRas and Akt in glial progenitor cells. To determine the reliance of these tumors on continued KRas signaling in vivo, we generated a viral vector that allows the expression of KRas to be controlled post-delivery. Tumor-free survival rates were compared between those animals with continued KRas expression and animals in which KRas expression was suppressed. KRas signaling was found to be required for the maintenance of these tumors in vivo; inhibition of KRas expression resulted in apoptotic tumor regression and increased survival. Subsequent reexpression of KRas reinitiated tumor growth, indicating that a percentage of the progenitor cells survived and retained tumorigenic properties.     

Examination of the in vitro (anti)estrogenic, (anti)androgenic and (anti)dioxin-like activities of tetralin, indane and isochroman derivatives using receptor-specific bioassays

Molecules derived from tetralin, indane and isochroman are often used in the synthesis of fragrance materials. The two polycyclic musk fragrances AHTN (6-acetyl-1,1,2,4,4,7-hexamethyltetralin), HHCB (1,2,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-gamma-2-benzopyran) and ADBI (4-acetyl-1,1-dimethyl-6-tert-butylindane) are derived from tetralin, isochroman and indane, respectively. In previous studies, AHTN and HHCB have been shown to antagonize estrogen receptors (ERs), both in vitro and in vivo. Here, we used two newly developed reporter gene assays, to examine the agonistic and antagonistic properties of several indane, tetralin and isochroman derivatives towards the human androgen receptor (AR) and aryl hydrocarbon receptor (AhR). Additionally, we also assessed (anti)estrogenicity of these compounds. A number of compounds showed weak estrogenic activity towards the human ER alpha. Several compounds showed (anti)estrogenic effects, starting at a concentration of 0.1 microM. Surprisingly, almost all compounds were found to be AR antagonists, starting at 0.1 microM. None of the compounds tested, showed either agonism or antagonism towards the AhR. Non-specific effects via crosstalk of the AhR and the ER or AR can therefore be ruled out. As far as we are aware, molecules derived from indane, tetralin and isochroman showing direct interaction with the ER and AR have not been reported previously.     

Individual differences in drug dependence in rats: the role of genetic factors and life events

Drug dependence and addiction is a chronic mental illness that has far reaching consequences for society in terms of economic loss, health costs and judicial problems. A crucial question in drug addiction, is what factors are involved in its aetiology, and especially what mediates the shit from use to abuse. As in most other mental illnesses, addiction can best be described using the so-called three hit model, which states that a disease results from an interaction between genetic factors, early lie events and late environmental factors. However, the precise nature of these factors still remains to be elucidated. This present review discusses the results from an animal model in which these three different hit are currently being investigated. The apomorphine susceptible (APO-SUS) and apomorphine unsusceptible (APO-UNSUS) rats, originally selected on the basis of their behavioural response to the dopaminergic agonist apomorphine, were recently found to be genetically different in the number of gene copies of a component of the gamma-secretase complex called Aph-1b. Whereas APO-UNSUS rats have three copies of the gene, APO-SUS rats have either 1 or 2 copies. In addition we have shown that these rats show differences in cocaine and alcohol self-administration, and that both early life events and late environmental factors can alter this self-administration behaviour. Thus, the data so far support the hypothesis that the APO-SUS and APO-UNSUS rats offer an interesting animal model for drug dependence in which genes and environment interact. We finally propose a theoretical model which can explain this gene-environment interaction.     

Hydrochlorothiazide increases plasma or tissue angiotensin-converting enzyme-inhibitor drug levels in rats with myocardial infarction: differential effects on lisinopril and zofenopril

Sodium depletion with diuretics augments the efficacy of angiotensin-converting enzyme-inhibitor therapy for hypertension and renal dysfunction, and possibly for left ventricular dysfunction after myocardial infarction. Underlying mechanisms may involve altered angiotensin-converting enzyme-inhibitor pharmacokinetics. We hypothesized that the diuretic hydrochlorothiazide causes increased steady-state levels of the angiotensin-converting enzyme-inhibitors lisinopril and zofenopril in rats with myocardial infarction. Rats were subjected to coronary ligation to induce myocardial infarction. After 1 week, rats were randomized to 50 mg/kg/day hydrochlorothiazide or control treatment for 3 weeks. The last week, rats received lisinopril or zofenopril in equipotentent dosages (3.3 and 10 mg/kg/day, respectively). Rats were sacrificed at Tmax after the last dose of angiotensin-converting enzyme-inhibitor, and tissues were collected for analysis of drug concentrations. Lisinopril concentrations in plasma were significantly increased by hydrochlorothiazide, at unchanged tissue concentrations. This increase could be fully explained by decreased renal function, as evidenced by increased plasma creatinine levels (lisinopril + hydrochlorothiazide 82+/-5 microM versus lisinopril 61+/-5 microM, P < 0.001). In contrast, zofenoprilat levels in kidney and non-infarcted left ventricle were markedly increased by hydrochlorothiazide, whereas plasma concentrations were unchanged. Although hydrochlorothiazide tended to increase plasma creatinine in zofenopril-treated rats as well, this increase was less pronounced (zofenopril + hydrochlorothiazide 61+/-3 microM versus zofenopril 54+/-2 microM, P = 0.15). Hydrochlorothiazide increases steady-state angiotensin-converting enzyme-inhibitor drug levels, most likely by affecting their renal clearance. Notably, the lipophilic angiotensin-converting enzyme-inhibitor zofenopril accumulated in tissue, whereas the hydrophilic lisinopril increased in plasma. Whether combining different angiotensin-converting enzyme-inhibitors with hydrochlorothiazide translates into distinct clinical profiles requires further study.     

Formation of transient covalent protein and DNA adducts by quercetin in cells with and without oxidative enzyme activity

This study investigates the role of cellular tyrosinase and/or peroxidase-like oxidative enzyme activity in the covalent binding of quercetin to glutathione, protein, and DNA, as well as the stability of quercetin DNA adducts in time. This was done by studying the formation of glutathionyl quercetin adducts in various in vitro models, and the covalent binding of radiolabeled quercetin to protein and DNA in cells with elevated peroxidase or tyrosinase levels and in cells devoid of nucleotide excision repair (NER). Cells with elevated tyrosinase or peroxidase levels contained approximately 2 times higher levels of covalent quercetin adducts than cells without detectable levels of these oxidative enzymes. However, this difference was smaller than expected based on the differences in tyrosinase and/or peroxidase levels, indicating that these types of oxidative enzyme activities do not play a major role in the cellular pro-oxidant activity of quercetin. Furthermore, quercetin DNA adducts were of transient nature, independent of the presence of NER, suggesting chemical instability of the adducts. Whether this transient nature reflects real reversibility or formation of genotoxic, depurinated sites remains to be investigated at the molecular level. Together, these data indicate that formation of covalent quercetin adducts can be expected in all cells, independent of their oxidative enzyme levels, whereas the transient nature of the DNA adducts formed may limit or cause their ultimate biological impact. If the transient nature represents chemical reversibility of the adduct formation, it would provide a possible explanation for the apparent lack of in vivo carcinogenicity of this in vitro mutagen. Therefore, in vitro mutagenicity studies should focus more on the transient nature of DNA adducts responsible for the mutagenicity in vitro, since this transient nature of DNA adducts may play an essential role in whether the genotoxicity observed in vitro will have any impact in vivo.     

Out of hospital treatment of patients with melioidosis using ceftazidime in 24 h elastomeric infusors, via peripherally inserted central catheters

BACKGROUND: In the tropical north of the Northern Territory, Australia, 25-50 patients are admitted to Royal Darwin Hospital (RDH) each year with Burkholderia pseudomallei infection, or melioidosis. Treatment consists of initial intensive therapy with 2-4 weeks of intravenous antibiotics. Clinical improvement may occur early and patients often prefer to be managed out of hospital in the Hospital in the Home (HITH). OBJECTIVES: To evaluate safety and efficacy of HITH management of patients with melioidosis. METHODS: A prospective observational study of our standard management which consists of 24 h infusions of ceftazidime infused through a peripherally inserted central catheter (PICC) line, plus oral sulphamethoxazole trimethoprim. Treatment is administered in the home, which may be in Darwin, regional areas or remote communities, or in a self-care unit located in the hospital grounds. RESULTS: From February 1998 to December 2001 150 patients were admitted to RDH with culture confirmed B. pseudomallei infection. Of these, 73 patients were treated with 24 h infusions of ceftazidime, of which 70 patients were managed by HITH. Complications of treatment include a PICC line complication rate of 10.6/1000 days in situ. Nine patients had relapse or recrudescence of disease, nearly all as a result of poor adherence to subsequent oral eradication therapy, these patients were all re-treated successfully. One patient remains infected with B. pseudomallei. CONCLUSION: This clinical outcome study suggests that out of hospital management of melioidosis with 24 h infusions of ceftazidime via a PICC line is safe and effective.