A Phase II trial of axitinib in patients with various histologic subtypes of advanced thyroid cancer: long-term outcomes and pharmacokinetic/pharmacodynamic analyses.

PURPOSE: Axitinib, a potent and selective second-generation inhibitor of vascular endothelial growth factor receptors, has shown activity in advanced thyroid cancer in a Phase II study. We report updated overall survival and pharmacokinetic/pharmacodynamic (PK/PD) analyses from the study. METHODS: Patients (N = 60) with advanced thyroid cancer of any histology for whom iodine-131 ((131)I) failed to control the disease or (131)I was not appropriate therapy were administered axitinib 5 mg twice daily. Objective response rate (primary endpoint), duration of response, progression-free survival, overall survival, safety, and PK/PD relationships were assessed. RESULTS: Objective response rate was 38 % [23 partial responses; 95 % confidence interval (CI) 26-52], and 18 (30 %) patients had stable disease lasting ≥16 weeks. Responses occurred in all histologic subtypes. With median follow-up of 34 months (95 % CI 32-37), median overall survival was 35 months (95 % CI 19-not estimable), median progression-free survival was 15 months (95 % CI 10-20), and median duration of response was 21 months (95 % CI 13-46). Most common Grade 3/4 treatment-related adverse events included hypertension (13 %), proteinuria (8 %), diarrhea (7 %), weight decrease (7 %), and fatigue (5 %). PK/PD analyses revealed trends toward greater tumor size reduction and response probability with higher axitinib plasma exposures. CONCLUSIONS: Axitinib appears active and well tolerated in patients with various histologic subtypes of advanced thyroid cancer, demonstrating durable responses and long overall survival. Axitinib may be useful for the treatment of advanced thyroid cancer.

Randomised phase II study of axitinib or bevacizumab combined with paclitaxel/carboplatin as first-line therapy for patients with advanced non-small-cell lung cancer.

BACKGROUND: Efficacy and safety of first-line axitinib/paclitaxel/carboplatin versus bevacizumab/paclitaxel/carboplatin in advanced non-squamous non-small-cell lung cancer (NSCLC) was evaluated. PATIENTS AND METHODS: Patients with stage IIIB/IV disease stratified by adjuvant therapy and gender were randomised 1 : 1 to axitinib (5 mg twice daily) or bevacizumab [15 mg/kg every 3 weeks (Q3W)], both with paclitaxel (200 mg/m(2) Q3W)/carboplatin (AUC 6 mg min/ml Q3W). RESULTS: The trial was discontinued after preliminary analysis. Median progression-free survival (primary end point) for axitinib (N = 58) and bevacizumab (N = 60), respectively, was 5.7 and 6.1 months [hazard ratio (HR) 1.09, 95% confidence interval (CI) 0.68-1.76; one-sided stratified P = 0.64]; median overall survival was 10.6 and 13.3 months (HR 1.12, 95% CI 0.74-1.69; one-sided stratified P = 0.70). Objective response rates (95% CI) were 29.3% (18.1-42.7) and 43.3% (30.6-56.8), respectively; risk ratio 0.676 (95% CI 0.41-1.11; one-sided stratified P = 0.94). The most common grade 3/4 adverse events included neutropenia (28% versus 20%), fatigue (14% versus 7%), and hypertension (14% versus 5%). Patient-reported outcomes based on the EORTC QLQ-C30 were similar between arms. CONCLUSIONS: In patients with advanced non-squamous NSCLC, axitinib/paclitaxel/carboplatin did not improve efficacy versus bevacizumab/paclitaxel/carboplatin, and was less well tolerated.

A phase III randomized study of gemcitabine and cisplatin with or without PF-3512676 (TLR9 agonist) as first-line treatment of advanced non-small-cell lung cancer.

BACKGROUND: This open-label phase III study assessed the addition of Toll-like receptor 9-activating oligodeoxynucleotide PF-3512676 to gemcitabine/cisplatin chemotherapy in patients with non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Chemotherapy-naive patients with stage IIIB or IV NSCLC were randomized (1:1) to receive six or fewer 3-week cycles of i.v. gemcitabine (1250 mg/m2 on days 1 and 8) and cisplatin alone (75 mg/m2 on day 1, control arm) or combined with s.c. PF-3512676 0.2 mg/kg on days 8 and 15 of each chemotherapy cycle and weekly thereafter until progression or unacceptable toxicity (experimental arm). No crossover was planned. The primary end point was overall survival (OS). RESULTS: A total of 839 patients were randomized. Baseline demographics were well balanced. Median OS (11.0 versus 10.7 months; P=0.98) and median progression-free survival (PFS) (both 5.1 months) were similar between groups. Grade≥3 hematologic adverse events (AEs), injection-site reactions, and influenza-like symptoms were more frequently reported among patients receiving PF-3512676. At the first-interim analysis, the Data Safety Monitoring Committee recommended study discontinuation. Administration of PF-3512676 was halted based on efficacy futility and increased grade≥3 AEs (experimental arm). CONCLUSIONS: Addition of PF-3512676 to gemcitabine/cisplatin chemotherapy did not improve OS or PFS but did increase toxicity.

Different subcellular localization and phosphoinositides binding of insulin receptor substrate protein pleckstrin homology domains.

Insulin evokes diverse biological effects through receptor-mediated tyrosine phosphorylation of the insulin receptor substrate (IRS) proteins. Here, we show that, in vitro, the IRS-1, -2 and -3 pleckstrin homology (PH) domains bind with different specificities to the 3-phosphorylated phosphoinositides. In fact, the IRS-1 PH domain binds preferentially to phosphatidylinositol 3,4,5-trisphosphate (PtdIns-3,4,5-P3), the IRS-2 PH domain to phosphatidylinositol 3,4-bisphosphate (PtdIns-3,4-P2), and the IRS-3 PH domain to phosphatidylinositol 3-phosphate. When expressed in NIH-IR fibroblasts and L6 myocytes, the IRS-1 and -2 PH domains tagged with green fluorescent protein (GFP) are localized exclusively in the cytoplasm. Stimulation with insulin causes a translocation of the GFP-IRS-1 and -2 PH domains to the plasma membrane within 3-5 min. This translocation is blocked by the phosphatidylinositol 3-kinase (PI 3-K) inhibitors, wortmannin and LY294002, suggesting that this event is PI 3-K dependent. Interestingly, platelet-derived growth factor (PDGF) did not induce translocation of the IRS-1 and -2 PH domains to the plasma membrane, indicating the existence of specificity for insulin. In contrast, the GFP-IRS-3 PH domain is constitutively localized to the plasma membrane. These results reveal a differential regulation of the IRS PH domains and a novel positive feedback loop in which PI 3-K functions as both an upstream regulator and a downstream effector of IRS-1 and -2 signaling.

DPOAE modifications induced by pure tone overstimulation in guinea pigs.

Distortion Product Otoacoustic Emissions (DPOAEs) shows a vulnerability to acoustic overstimulation that is easily detectable by changes in amplitude, frequency distribution and time course. The frequency-specific changes in DPOAE provide more information than the more general changes in click evoked otoacoustic emissions. In this work five anesthetized guinea pigs were examined for changes DPOAE after exposure to pure tones. The noise was a 110 dB SPL pure tones for 45 minutes and the fatiguing tone centered on the geometric mean (GM) of primaries or 2/3 of GM. The most measurable effects were obtained in the latter conditions. The main DP level reduction lasts about one hour after exposure, but the complete recovery is observable only after 24 hours. The frequency distribution of fatiguing effects on the DP- audiogram shows a remarkable fine tuning and a pattern like a low-pass filter. After four hours, in one guinea pig, it is observable an enhancement of the DP amplitude, compared to the pre-exposure level; in another guinea pig, a second overstimulation produced a more extensive and time-lasting effects than the first exposure.

Zidovudine and thymus humoral factor gamma-2 in the treatment of HIV infection: preliminary clinical experience.

A case-control, prospective, open-label, clinical trial to evaluate efficacy and safety of a combined zidovudine/Thymus Humoral Factor Gamma-2 (THF) therapy in HIV-infected subjects was conducted in 13 patients. Twenty-six patients were included as controls receiving only zidovudine. The two groups of patients were matched according to sex, age, CDC stage of HIV infection, number of CD4+ T cells and type of previous opportunistic infections (if any) and all patients and controls were naive for antiretroviral therapy at the moment they entered the trial. The observation period was protracted up to 47 months (mean 28 +/- 13 months). No significant difference was observed between the two groups as far as surrogate markers of HIV disease progression are concerned. However, patients receiving zidovudine and THF showed a lower number of opportunistic complications. Only one patient in this group progressed to manifest AIDS while 9 of 18 controls presented disease progression. Four patients died in the case group, all of them were CDC stage IV at admission, and 15 of 26 died in the control group (all CDC stage IV at admission, and four patients who presented disease progression during the study period). Survival time was increased in the case group. The exact immunological effect of thymus hormones in HIV infection has still to be elucidated, but a possible therapeutic role of these agents is foreseeable.

Pachydermoperiostosis: dermatological, neurological and radiological observations.

A case of pachydermoperiostosis is described. Interesting features were the presence of carpal and tarsal tunnel syndromes, chronic leg ulcerations and large calcification of the Achilles tendon. Neurological alterations were explained by the stenosis of the tunnels secondary to the periosteal apposition. Chronic leg ulcerations were probably due to neurological and circulatory alterations.

Vision screening of preschool children in Italy.

Vision screening of preschool children is designed and performed to identify those affected by amblyopia or related, predisposing visual defects. In order to determine the prevalence of preschool vision screening in Italy, a questionnaire was mailed to the 635 regional health offices in which the country's national health system was organized at the time of the study. Results of this survey demonstrated that in 61.3% of the regions which responded, some form of preschool vision screening is performed. However, individual, non-standardized methods are used by the physicians or health care professionals responsible in each regional program. It is evident from this study that standardized procedures for preschool vision testing are lacking in Italy, and that the existing European Community guidelines should be disseminated and applied on a greater scale for the development of such a program on the national and European level.

Effect of oestrogen replacement on bone metabolism and cytokines in surgical menopause.

The effect of oestrogen replacement on bone metabolism and serum cytokine levels (IL1,IL6) was investigated in surgical menopause. The study included 40 female subjects; 10 healthy premenopausal women underwent total hysterectomy without oophorectomy. Thirty healthy premenopausal women underwent total hysterectomy with bilateral oophorectomy. They were randomly divided into 3 groups of 10 subjects. The first group received natural estradiol (0.05 mg/day) for 6 months; the second group received natural estradiol (0.05 mg/day) and medroxyprogesteron acetate (10 mg/day) for 6 months, the third group received no therapy. Calcium-phosphorus metabolism, inflammatory indices, serum IL1 and IL6 levels were tested before and 6 months after surgery in all patients. A significant increase in serum alkaline phosphatase, urinary cross-links, serum PTH and IL1-IL6 was observed in the untreated women with total hysterectomy and oophorectomy. No significant variation in any of the parameters considered was observed in patients treated with oestrogen, in those treated with oestrogens and medroxyprogesteron nor in patients without oophorectomy. These results in human "in vivo" confirm that ovarian steroids play an important role in regulating the production of IL1 and IL6 which could regulate bone resorption.

[Corneal transplantation in Parma: a critical review of activities from 1983 to 1989]. / Il trapianto di cornea a Parma: revisione critica dell'attività svolta dal 1983 al 1989.

At the Institute of Ophthalmology of Parma 66 corneal transplants were performed from 1983 to 1989. In this period the number of grafts per year remarkably increased. Two therapeutic choices characterized corneal transplantation in Parma: a) a fresh donor cornea was used without any storage medium; b) the post-operative ambulatory follow-up of patients was particularly frequent in the first 12 months after surgery. The follow-up period ranged from 3 to 75 months (mean 25 months). The life table calculated using the Kaplan-Meier method shows a graft survival of 86% after 4 years. Graft failures were 8. The authors report all postoperative complications that appeared. In comparison with what reported in Literature, a higher percentage of graft survival and a better final visual acuity were observed in the patients who underwent corneal transplantation in the Eye Clinic of Parma.

An exponential-Gompertzian description of LoVo cell tumor growth from in vivo and in vitro data.

The data of nine monolayer cultures, 48 multicellular spheroids, and 19 s.c. xenografts of LoVo cells were fitted, on an individual basis, by exponential and Gompertzian equations, respectively. The mean growth parameters alpha 0 (initial growth rate) and beta (retardation factor) of the three experimental systems presented a strong linear correlation alpha 0 = 6.88 beta + 0.56, r = 0.9843. This implies that, at the particular tumor size of 155 microns in diameter (mean +/- SD = 50-310 microns), the tumor growths described by Gompertzian curves (from spheroids as well as from s.c. xenografts) have the same growth rate as monolayer cultures. This occurrence strongly supports an exponential-Gompertzian growth model, where an exponential monolayer-like phase changes to a Gompertzian growth, controlled by environmental conditions, when the tumor size has reached a critical value.

Changes in the three-dimensional organization of LoVo cells associated with resistance to doxorubicin.

LoVo cells and a derived subline resistant to doxorubicin were compared in the spheroids system. The resistant line, unlike the parent one, was unable to grow as spheroids, but formed irregular loose aggregates. Moreover treatment of the resistant cells with membrane-active agents able to reverse pleiotropic drug resistance had no effect on the capability of these cells to grow as spheroids. The results indicate that the inability of resistant cells to form spheroids is not related to the resistance mechanism.

Malignant transformation of host cells by a human small cell lung cancer xenografted into nude mice.

Malignant transformation of mouse host cells by a human small cell lung cancer (SCLC) was demonstrated by short-term in vitro cultivation of the tumor cells from a xenograft at two different transplant generations. Isoenzyme (LDH) and chromosome analysis showed that out of the 3 cell lines established from this tumor, 1 retained a human karyotype similar to that of the xenograft and 2 were murine transformed cell lines. These murine cell lines produced fibro-sarcoma-like tumors when injected into nude mice. Because of the early in vitro emergence of murine transformed cell populations, it is likely that the transformation process had occurred in vivo. Since in our experience the induction of transformation of host murine cells, also observed directly in vivo, is more frequent with SCLC than other histotypes (lung and colorectal adenocarcinoma), it is suggested that the known production of growth factor by these tumors may contribute to this transformation.

A comparative study of the effects of anthracycline derivatives on a human adenocarcinoma cell line (LoVo) grown as a monolayer and as spheroids.

The cytotoxicity of doxorubicin (DX), 4'-O-methyl-DX (MET-DX), 4'-deoxy DX (DEO-DX), 4'-deoxy-4'-iodo-DX (IODO-DX), daunorubicin (DNR)and 4-demethoxy-DNR (DM-DNR) on LoVo cells cultured as a monolayer (in exponential and stationary phases of growth) and as spheroids, are evaluated following 1-h exposure to the drugs. All compounds were more cytotoxic than DX in both systems. A comparison of the ID50 values for cell survival of monolayer cells and of the inhibition of relative growth of individual spheroids indicated that MET-DX and DNR, like DX, were less cytotoxic on spheroids than on monolayer cells in both growth conditions; DEO-DX, IODO-DX and DM-DNR showed an activity on spheroids that was intermediate between that observed on monolayer cells in exponential and stationary phases of growth. This observation indicated that the different pattern of activity of these anthracyclines on a spheroids system is not only related to the presence of cells at different cell cycle phases, but presumably also depends on factors related to the 3-dimensional structure of spheroid cells. Indeed DEO-DX, IODO-DX and DM-DNR, more lipophilic than the other compounds, might penetrate more deeply into spheroids, thus improving the cytotoxic response in this experimental system.

Formation, growth and morphology of multicellular tumor spheroids from a human colon carcinoma cell line (LoVo).

LoVo human colon carcinoma cells cultured by a liquid overlay technique form and grow as multicellular tumor spheroids. The growth curve of LoVo spheroids exhibits Gompertzian growth kinetics, i.e., exponential growth for 10 days, followed by exponential retardation in the rate of growth. Doubling time in the exponential growth phase is longer than in monolayer cultures (5 days for LoVo spheroids vs. 37 h for monolayers). When LoVo spheroids reach a diameter of about 300 microns, a necrotic core appears in their center and continuously increases with spheroid growth. The cell ultrastructure and organization in spheroids closely resemble those of the same cells when grown as tumors in vivo or as monolayer, i.e. intestinal epithelium, desmosomes, intracytoplasmic lumina and acinar structures. Individual cells from spheroids can be obtained by trypsinization and assayed for colony formation. LoVo spheroids provide a model which can be readily manipulated and appears to be suitable for evaluation of anticancer drugs. A comparison of LoVo spheroids exposed to doxorubicin with the same cells grown in monolayers emphasized the role of cell organization in determining drug resistance.

Biological activity of metal complexes. I. Interaction of pt(II), pd(II) and rh(I) complexes with E. coli strains and with mice LS fibroblasts in vitro.

The effects of a number of Pt(II, Pd(II) and Rh(I) complexes against cultures of Escherichia coli (strains B, H10178, uvra-, recA-) and cultures of mice LS Fibroblasts were tested. Most of the compounds showed higher cytotoxic activity than the cis-Pt(NH3)2Cl2, the compound at present on clinical trial as antittumour drug. A new model of active compound is proposed.