Activity profiling of deubiquitinating enzymes in cervical carcinoma biopsies and cell lines

Ubiquitin specific proteases (USPs) regulate the production and recycling of ubiquitin and are thereby critically involved in the control of cell growth, differentiation, and apoptosis. Increasing evidence implicates deregulation of USPs in malignant transformation but there is very little information on the overall and specific activity of USPs in normal and tumor tissues. We have used a chemistry-based functional proteomics approach to profile the activities of individual USPs in biopsies of human papillomavirus (HPV) carrying cervical carcinoma and adjacent normal tissue. To assess the contribution of HPV proteins, USP activity was also compared in HPV positive and negative cervical carcinoma cell lines and HPV E6/E7 immortalized human keratinocytes. The activity of the C-terminal hydrolases UCH-L3 and UCH37 was upregulated in the majority of tumor tissues compared to the adjacent normal tissues. UCH-L1 activity was lower in a significant proportion of the tumors but to a less extent in advanced tumors. In accordance with the relatively low UCH-L1 activity in tumor biopsies, UCH-L1 was detected only in one out of eight cervical carcinoma lines. UCH-L1, UCH-L3, USP7, and USP9X activity was upregulated following HPV E6/E7 immortalization of keratinocytes, suggesting a role of these enzymes in growth transformation.     

Alternating chemo-immunotherapy with temozolomide and low-dose interleukin-2 in patients with metastatic melanoma

Temozolomide is a rapidly absorbed chemotherapeutic agent, achieving significant central nervous system penetration. Previous clinical trials suggested that temozolomide in sequence with low-dose recombinant human interleukin-2 might be an efficacious and relatively non-toxic chemo-immunotherapeutic treatment, which may synergistically eliminate tumours. The primary objective was to determine the safety and tolerance of temozolomide administered orally 200 mg/m days 1-5, in sequential combination with subcutaneous injections of 4.5x10 IU recombinant human interleukin-2 on days 8-11, 15-18 and 22-25 in patients with measurable, progressive metastatic malignant melanoma without radiological signs of central nervous system metastases. The secondary objectives were to determine tumour response and time to progression. Twenty-seven patients were included, of which four were non-evaluable for response. Twenty-three patients tolerated the regimen with side effects below grade 3 according to the World Health Organization (WHO) scale. Three patients suspended the treatment because of WHO grade 3 side effects already during the first 3 days of the first course of temozolomide. Seven patients showed no tumour progression during the first four treatment cycles. Two patients had complete responses, three partial responses and two stable disease at the end of the four cycles defined by the protocol and they continued the treatment until signs of relapse or a maximum of 21 courses. Five of these patients are still alive. Thrombocytopenia was significantly more pronounced in patients with objective response and stable disease than in non-responders to therapy. The median time to progression for all patients was 3.1 months and for responding and stable disease patients was 15 months. Five of 23 treated patients (22%) developed brain metastases during follow-up. Temozolomide in combination with recombinant human interleukin-2 is a well-tolerated regimen for outpatient treatment and the bio-chemotherapy combination induced durable clinical responses. Thrombocytopenia might be a positive predictive factor for response to therapy.     

The Tumor Promoter Phorbol-12,13-Dibutyrate [P(BU)2] Stimulates Cytotoxic Activity of Human Blood Lymphocytes

The tumor promoters 12-13-phorbol-dibutyrate, P(Bu)2, and 12-0-tetradecanoylphorbol13-acetate, TPA, were shown to augment the cytotoxic potential of human blood lymphocytes with low cell density. In kinetics experiments the enhancing effect was preceded by an initial suppression lasting for about 2 hours. Admixture of mononuclear adherent cells abrogated the P(Bu)2 effect in a dose dependent way. P(Bu)2 altered the sensitivity of K562 cells to the cytotoxic effect. Short term pretreatment increased the sensitivity, but after longer pretreatment the cells became resistant. The results show that tumor promoters can influence the cytolytic system at different levels. By acting directly on the lymphocytes they potentiate the lytic function. When mixed mononuclear populations are used, this effect may be counteracted via activation of the suppressive functions of monocytes. In addition, the target cell sensitivity can also be modulated. As a result, the final outcome of phorbol treatment depends on the strength, kinetics and the mode of its effects on the interactants.     

Adrenal glands in patients with congenital renal anomalies: CT appearance

The CT appearance of the adrenal glands was investigated in 30 patients with congenital renal anomalies (17 cases of unilateral renal agenesis, 11 of inferior ectopy, and 2 of crossed fused ectopy). The ipsilateral adrenal was clearly identified in 83% of these patients; in all of them, the adrenal was a paraspinal disk-shaped organ, which appeared linear on CT. Conversely, the adrenals retained their normal shape in a control group of 20 patients with acquired renal atrophy or prior simple nephrectomy.