Autoradiographic analysis of 5-HT2A binding sites in the brain of Sardinian alcohol-preferring and nonpreferring rats.

The density of 5-HT2A binding sites in the brain of Sardinian alcohol-preferring (sP) and nonpreferring (sNP) rats was evaluated, using [3H]ketanserin for quantitative autoradiography. The highest [3H]ketanserin binding levels were found in the anterior olfactory nucleus, prefrontal cortex, medial prefrontal cortex, post-genual anterior cingulate cortex, insular cortex and claustrum. Statistically significant differences between sP and sNP rats were found in prefrontal cortex, medial prefrontal cortex and post-genual anterior cingulate cortex, where sP rats showed about 20% lower [3H]ketanserin binding levels. No significant difference was found in other areas, although some of them showed slightly lower [3H]ketanserin binding density in sP rats. The 5-HT2A receptor agonist, (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-amino-propane hydrochloride (DOI), microinjected into the medial prefrontal cortex, induced a lower number of wet dog shakes in sP than in sNP rats. These results indicate a different density of 5-HT2A binding sites, and a different functional regulation of 5-HT2A receptor mechanisms in discrete brain areas of sP, in comparison to sNP rats. These findings, and those showing lower levels of 5-HT in the frontal cortex of sP rats, suggest that altered 5-HT function in fronto-cortical areas could be linked to the genetic predisposition to high voluntary ethanol intake in these rats.     

Salvage therapy with ilosfamide and mitoxantrone in advanced ovarian cancer

OBJECTIVE:: To evaluate the anti-tumour activity and toxicity of ifosfamide(5 g/m² continuous infusion) and mitoxantrone (10 mg/ m² givenin combination every 3 weeks in patients with ovarian cancerresistant to at least two previous regimens which included platinum. PATIENTS AND METHODS:: Additional eligibility criteria were an ECOG performance status2 and measurable disease. Of 47 patients entered in the study,8 were defined as platinum-resistant and 39 as potentially sensitiveaccording to Markman's criteria. Thirty-five patients had alsoreceived pacitaxel as last treatment before entering this study.Tumour response was evaluated every three cycles. RESULTS:: One complete and 11 partial responses were reported, for anoverall response rate of 25% (95% CI: 14%–40%). Threeof the partial responders were resistant to num. None of the7 partial responders pretreated with had responded to it. Theoverall median urvival was months. Neutropema G4 was reportedin 18 patients(42%) with hospitalisation because of febrileneutropenia in 3 of them. CONCLUSIONS:: In patients with ovarian cancer failing least 2 previous therapiesincluding platinum, the nation of ifosfamide and mitoxantronehas shown antitumour activity comparable to that of paclitaxel,with accept able toxicity. Objective responses were reportedalso patients failing pacitaxel, suggesting a lack of crossresistance. ifosfamide, mitoxantrone, ovarian cancer     

Brachytherapy for isolated vaginal recurrences from endometrial carcinoma

AIMS AND BACKGROUND: Isolated vaginal recurrences of endometrial carcinoma are rare, and prognostic factors that predict treatment outcome are still not well defined. The aim of the present study was to evaluate the results of brachytherapy in isolated vaginal recurrences from endometrial carcinoma. METHODS: Thirty-five patients with isolated vaginal recurrences were treated with brachytherapy with intravaginal ovoids or cylinders that were calculated to deliver 6000 to 7000 cGy at the surface. Patients were assessed for size and location of recurrence at presentation, response and complications from therapy. RESULTS: Treatment was well tolerated by most patients. Grade 2 toxicity occurred in 4 patients (3 cases of partial vaginal stenosis and one proctitis). Complete response to radiation was observed in all patients, and an overall 9 failures were observed (4 local, 4 distant and 1 local plus distant). Twenty patients (57%) were alive without evidence of disease at 3 to 11 years following treatment. Site of vaginal recurrence (upper third versus others) and long (more than 12 months versus less than 12 months) interval from hysterectomy were the only factors significantly related to local failures. CONCLUSIONS: Isolated vaginal recurrences following hysterectomy for endometrial carcinoma can be treated with brachytherapy with a low rate of severe toxicity.     

Phase I and pharmacologic evaluation of intraperitoneal 5-fluoro-2′-deoxyuridine

Summary Intraperitoneal (i.p.) 5-fluoro-2-deoxyuridine (Floxuridine, FUdR, FdUrd) was evaluated in a phase I study at a starting level of 500 mg given on 1 day in 2 I 1.5% dialysate. Escalations within patients were allowed every other cycle. A total of 23 patients (age, 32–78 years) received 108 treatment courses. Local tolerance at all dose levels was excellent, with no cases of drug-related peritonitis being observed. Nausea and vomiting increased in severity in relation to dose and was universal at >3,000 mg ×3 days. One patient each developed grade 1 mucositis as well as diarrhea at a dose of 3,000 mg×3 days and leukopenia and thrombocytopenia at 5,000 mg×3 days. Peritoneal fluid (PF) and plasma (PL) FdUrd profiles were monitored by an HPLC method in 13 subjects, with 7 being studied serially at 2–4 increment doses for up to 6 h. Profiles that exhibited apparent linear pharmacokinetics gave PF drug levels 2–4 logs higher than the PL counterparts, with the latter essentially declining in parallel to the former, indicating that the disposition of FdUrd from the peritoneal compartment is rate-determining. The mean terminal half-life for PF FdUrd was found to be 115 min and mean peritoneal clearance was 25 ml/min. The vast differences in drug levels and AUC found between the PF and the PL profiles suggests a high systemic clearance of FdUrd, which was confirmed in two patients receiving 2 g FdUrd by short i.v. infusion. A disproportionate increase in the plasma FdUrd levels and the corresponding AUC values was found with increasing dose, suggesting a disproportionate increase in the systemic partitioning of FdUrd when doses were escalated within a patient. Substantial levels of peritoneal 5-fluorouracil (FUra) were also detected in most of the subjects. Thus, FdUrd was found to have several desirable properties for i.p. administration: (1) a 2- to 4-log pharmacologic advantage, (2) the absence of local toxicities, and (3) a favorable antitumor spectrum and some evidence of antitumor effects in this phase I and pharmacology study. A 3,000-mg dose given in 2 l 1.5% dialysate for 3 consecutive days exhibited antitumor activity and produced no systemic toxicity except nausea and vomiting, which was controlled by antiemetics. This dose schedule is therefore recommended for phase II trials directed against small-volume disease in the peritoneal cavity, such as may be found in some stages of ovarian and gastrointestinal cancers. In addition, it is suitable for further exploration as a part of regimens including systemic therapy or drugs that modulate the action of fluoropyrimidines.Supported in part by Cancer Center Core Grant CA 14089, by ROI CA 50 412, by an ACS Institutional Grant (IN21Z, to C. R.) and by the Italian-American Foundation award (to N. C.)Deceased     

Treatment of the abdominal aorta combined with a second surgical procedure

The synchronous performance of aortic graft with a second surgical procedure should be avoided because of the risk of subsequent graft infection. The Authors reported the experience of 21 patients who underwent synchronous aortic graft with second surgical procedure. No graft infection occurred and it is concluded that such combined operation may be safer to perform than staged surgery in elderly patients.     

Zentralnervöse Kontrolle der Miktion bei Patienten mit Blasenfunktionsstörung im Vergleich zu gesunden Kontrollpersonen

PURPOSE: The pontine micturition center plays a central role in regulating the micturition reflex, but the precise neural mechanisms are unclear. The cerebral cortex is involved in coordinating micturition but there is little knowledge on specific evolutionary higher brain regions. The present study aimed to investigate whether cortical activation during micturition can be demonstrated by EEG power spectra patterns and to explore whether specific cortical regions involved in the interaction of inhibition and release during the micturition reflex can be discerned. We also aimed to test whether intravesical electrostimulation (IVES) therapy in patients with micturition disorders has an effect on patterns of cortical activity. METHODS: The healthy control group was divided into those who were able to void when requested (6 women, 12 men) and those who were not (8 women, 10 men). These subgroups were compared separately with the 14 patients before and after IVES for voiding dysfunction. Following IVES all patients were able to void spontaneously. Mean age of the patients and healthy volunteers was 52 and 30 years, respectively. At the beginning of the study all subjects had a bladder volume of approximately 250 mL as measured by sonography. The EEG was obtained at rest and during the attempt to void. In the patients' group EEG was obtained before IVES treatment and at the day of the last stimulation. The measurement period lasted about 6 minutes. At the beginning of the recording the proband was asked to close his/her eyes. During the resting period after 1 minute the patient was asked to open his/her eyes. After 10 seconds he/she was asked to close his/her eyes again. Then, with eyes still closed, the patient was asked to void. During the entire EEG recording the patient was seated in a comfortable, electrically isolated chair in a darkened room and separated from the examiner by a partition. The subject was asked to relax and not move his/her eyes. The EEG was recorded from the 19 standard points (10-20 System) versus an averaged mastoid electrode with a gold-plated cup electrode (Glass). An EOG was recorded simultaneously to register eye artefacts. The amplification chain was calibrated with a 10-Hz 100-microVss sinus signal generated with a biosignal amplifier. The transitional resistances of all EEG channels were less than 5 kOhm and established as soon as possible. EEG and EOG signals were amplified and recorded with a B.E.S.T. Brain Mapping System. The recording frequency was 256 Hz and the resolution of the analog digital conversion was 12 bit. A high pass and a low pass filter were set to 0.53 Hz and 70 Hz, respectively. All recordings were inspected visually before computer analysis. Artefacts were marked and excluded from the further analysis. None of the EEG recordings showed clinical abnormalities. As expected, the EEGs during voiding attempts showed some muscle potentials and slow motion artefacts. For each subject two artefact-free resting segments of about 20 seconds, one from the resting phase and one from the voiding attempt, were defined by hand for automated analysis. Relative power spectra (microV2) were calculated for the defined segments. From the spectra the relative alpha band power (7.5-13.0 Hz) was calculated for each subject for rest and voiding. Group (patients vs. voiding probands vs. probands unable to void) and sex were independent variables. The alpha power of the 17 electrode positions of the 10-20 system (without Fp1 and Fp2) during rest and attempted voiding were repeated measurement variables. The frontopolar electrode was not used because of its susceptibility to artefacts. The number of dependent variables was due to the explorative nature of the study. With interactions of variables with more than two factor levels a Greenhouse-Geisser correction was performed. Interactions were subjected to contrast analysis and Newman-Keuls-Post tests. RESULTS: Significant effects were seen for BEDINGUNG (     

p15INK4b, p14ARF, and p16INK4a inactivation in sporadic and neurofibromatosis type 1-related malignant peripheral nerve sheath tumors.

PURPOSE: Malignant peripheral nerve sheath tumor (MPNST) can arise sporadically or in association with neurofibromatosis type 1. Deletions at the 9p21 locus have been reported in these tumors. To additionally characterize the status of this chromosomal region, in this study we performed a comprehensive, mostly PCR-based molecular analysis of the three tumor suppressor genes p15(INK4b), p14(ARF) and p16(INK4a) located at the 9p21 locus in 26 cryopreserved MPNSTs. EXPERIMENTAL DESIGN: Fourteen neurofibromatosis type 1-related and 12 sporadic cases were investigated for homozygous deletion coupled with fluorescent in situ hybridization, promoter methylation, and mutational analysis, as well as m-RNA expression. RESULTS: The results showed that an inactivation of one or more genes occurred in 77% of MPNSTs and was mainly achieved through homozygous deletion (46%), which, in turn, encompassed all of the three tandemly linked genes in 83% of the deleted cases. Promoter methylation was at a less extent involved in gene silencing (18%), and no mutations were found. Loss of function at DNA level strongly correlated with loss of mRNA expression accounting for 80% of the cases. Because of the close relationship between p14(ARF) and TP53 and between p15(INK4b)/p16(INK4a) and Rb, these results support a model of a coinactivation of TP53 and Rb pathways in 75% of MPNSTs, with functional consequences on cell growth control and apoptosis. CONCLUSIONS: The inactivation of the 9p21 locus is a frequent and peculiar hallmark of MPNST genetic profile leading also to an impaired apoptosis that could be taken into account in treatment planning of these tumors.     

Enhancement of in vivo antitumor activity of classical anticancer agents by combination with the new, glutathione-interacting DNA minor groove-binder, brostallicin.

PURPOSE: Brostallicin (PNU-166196) is a alpha-bromoacrylic DNA minor groove binder, currently in clinical evaluation. This drug has the peculiarity of showing enhanced antitumor activity in cells with high glutathione S-transferase (GST)/glutathione content. The purpose of the study was to study multiple combinations of brostallicin with classical anticancer agents. EXPERIMENTAL DESIGN: The cis-dichloro-diammine-platinum (cDDP)/brostallicin combination was tested in the human colon carcinoma (HCT-116) model transplanted in nude mice. Two treatment schedules were tested: cDDP followed by brostallicin 48 h after or brostallicin followed by cDDP. These two schemes were selected from the observation that tumor cells in vitro show an increased activity of GST 48 h after cDDP treatment. The HCT-116 model was used also to test the irinotecan (cPT-11)/brostallicin combination. The effect of brostallicin in combination with doxorubicin (DX) was studied in the i.v. injected murine L1210 leukemia. Three administration schedules were tested. The antitumor activity of brostallicin and Taxotere was tested on the A549 lung cancer xenografts. RESULTS: In line with the increased GST activity observed after treatment with cDDP, the cDDP/brostallicin interaction was sequence-dependent, leading to a more than additive antitumor effect, without additional toxicity, only when cDDP was given before brostallicin. The antitumor effect of CPT-11 was enhanced significantly by brostallicin cotreatment. A more than additive antitumor effect, without additional toxicity, was observed when DX/brostallicin were sequentially administered in L1210-bearing mice. Finally, additivity was observed when brostallicin/Taxotere simultaneous combination was tested. CONCLUSIONS: Although the precise molecular mechanism of interaction between brostallicin and the other tested cytotoxics has not yet been identified, a clear therapeutic gain is observed in preclinical models when brostallicin is combined with anticancer agents such as cDDP, DX, CPT-11, and Taxotere. These results indicate the potential therapeutic value of brostallicin in cancer combination treatment therapy.     

Immunotherapy I: Cytokine gene transfer strategies

The cytokine approach to gene therapy of cancer stems from early studies of direct, repeated injection of recombinant cytokines at the tumor site, and extension of the bystander effect that enables a few cytokine gene transduced cells in a tumor to bring about its total destruction. This effect can be extended through the immune system, since cytokine-activated regression of a small mass of tumor cells can afford systemic protection. Transduced cells used as a vaccine provide a local concentration of both cytokine and tumor antigens. Cytokines sustain antigen uptake and presentation by increasing the immunogenic potential of the environment through the recruitment of antigen presenting cells and leukocytes, and activation of a cascade of events which amplify and tone up the efficacy of a vaccine. The promises and difficulties of this approach are discussed by considering what is still missing from experimental studies and what can best be done as soon as possible in animals and humans to reach compelling conclusions.     

Corrigendum

M. Maneo should read A. Maneo. The author's correct affiliationis III Clinica Ostetrico Ginecologica, Università diMilano, Milan, Italy.