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A calculated panel reactive antibody (cPRA) estimates the percentage of donors with unacceptable antigens (UA) for a recipient. cPRA may be underestimated in transplant candidates with UA to DQA, DPA, and DPB if these are not included in the calculation program. To serve the National Canadian Transplant Programs, a cPRA calculator was developed with complete molecular typing for all donors at HLA‐A, B, C, DRB1, DRB3/4/5, DQA1, DQB1, DPA1, and DPB1, all resolved to serologic equivalents. The prevalence of UA at DQA, DPA and DPB was evaluated in a sensitized regional population. The impact of adding these additional UA to cPRA was calculated alone and in combination, and compared to the baseline cPRA for UA at A, B, C, DR, DR51/52/53, and DQ. Of 740 sensitized transplant candidates, 18% of total and 32% with cPRA≥95% had DQA UA. Twenty‐seven percent of total and 54% with cPRA≥95% had DPB UA. Of 280/740 subjects with these UA, 36/280 (13%) had cPRA increase of >20% when they were included, 7% increased cPRA to ≥80% and 6% to ≥95%. Inclusion of DQA, DPA, and DPB UA in Canadian cPRA calculations improves the accuracy of cPRA where these are relevant in allocation., Inclusion of DQA, DPA and DPB in the Canadian cPRA calculator increases cPRA by >20% in 13% of patients with these unacceptable antigens, improving cPRA estimates where these loci are relevant in virtual crossmatching.

Introduction Nous rapportons un cas original d’erytheme pigmente fixe (EPF) au fulvestrant, confirme par patch-tests. Le fulvestrant est un agoniste des recepteurs aux œstrogenes, indique chez les patients atteints de cancer du sein metastatique hormonodependant. Aucun cas d’EPF au fulvestrant n’a ete rapporte dans la litterature. Discussion Une femme de 69 ans etait traitee depuis septembre 2016 par injections sous-cutanees de fulvestrant et denosumab (anticorps monoclonal diminuant la resorption osseuse) tous les 15 jours en alternance dans le cadre d’un cancer du sein metastatique. En janvier 2017, elle a presente des lesions erythemateuses, prurigineuses, et bien delimitees du tronc, avec pigmentation sequellaire, evocatrice d’EPF a l’examen clinique. Les lesions recidivaient regulierement, evoquant l’imputabilite du fulvestrant ou du denosumab. Il n’y avait pas d’autre medicament recemment introduit. La remission des lesions etait observee apres arret des deux medicaments. Des explorations allergologiques etaient effectuees en juin 2017. Les patch-tests au fulvestrant a la concentration de 250 mg/mL etaient positifs sur peau saine (dos) et sur peau lesee (region lombaire) a la lecture a 48 h et 96 h. Les patch-tests au denosumab a la concentration de 70 mg/mL etaient negatifs sur peau saine et peau lesee. L’examen histologique des tests positifs au fulvestrant mettait en evidence un infiltrat lichenoide avec necroses keratinocytaires. Le diagnostic d’EPF au fulvestrant etait retenu. Une declaration en pharmacovigilance etait effectuee. Aucune recidive des lesions n’etait constatee apres reprise du denosumab et remplacement du fulvestrant par l’exemestane (inhibiteur de la synthese des œstrogenes). Conclusion L’EPF est une forme peu frequente de toxidermie. Il s’agit du premier cas decrit d’EPF au fulvestrant, confirme par explorations allergologiques et analyse histologique.

Background Standard adjuvant chemotherapy regimens for patients with node positive (N+) breast cancer consisted of anthracycline followed by taxane. The European Association for Research in Oncology embarked in 2000 on a phase III trial comparing 6 cycles of FEC 100 versus 4 FEC 100 followed by 4 Taxol. Primary end-point was disease free survival. Secondary end-points were overall survival, local recurrence free interval, metastases free interval and safety. Patients and methods Between March 2000 and December 2002, 837 patients were randomised between 6FEC 100 for 6 cycles (417 patients) or FEC 100 for 4 cycles then Taxol 175 mg/m 2 /3 weeks for 4 cycles (4FEC 100 -4T) (420 patients). One thousand patients had been planned initially but the trial was closed earlier due to slow accrual. Results Hazard ratios (HRs) were 0.99 for disease-free survival (DFS) (95%CI: 0.77–1.26; p = 0.91), and 0.85 for overall survival (OS) (95%CI: 0.62–1.15; p = 0.29). Nine-year DFS were 62.9% versus 62.5% for 6FEC 100 and 4FEC 100 -4T, respectively. Nine-year OS were 73.9% versus 77% for 6FEC 100 and 4FEC 100 -4T, respectively. Toxicity analyses based on 803 evaluable patients showed that overall grade 3–4 toxicities were similar in both arms (63% versus 58% for 6FEC 100 arm and 4FEC 100 -4T arm, respectively; p = 0.16). Conclusion In this trial replacing the last 2 FEC 100 cycles of 6FEC 100 regimen by 4 Taxol does not lead to a discernable DFS or OS advantage. The lack of a significant difference between the randomised treatment arms may however be due to a lack of power of this trial to detect small, yet clinically worthwhile, treatment benefits.

We report the case of a woman with metastatic breast cancer receiving intrathecal trastuzumab (and intravenous trastuzumab for more than 7 years). She was diagnosed in 2001 with a duct invasive breast cancer T3N1M0 HER2 (human epidermal growth factor receptor 2)-positive +++ HR (hormone receptor) -negative. She received chemotherapy and then she had a mastectomy. Several metastases were discovered and treated from 2003 to 2008 with chemotherapy. In March 2010, brain metastases and a leptomeningeal carcinomatosis from her HER2-positive breast cancer appeared. From that moment on she received intravenous trastuzumab (6 mg/kg) every 3 weeks, intrathecal trastuzumab (21 mg) weekly for 16 injections and lapatinib. Intrathecal trastuzumab was stopped because of cerebrospinal fluid (CSF) clearing. Intrathecal trastuzumab was injected again from December 2013 for 14 injections. The relevance of treating leptomeningeal carcinomatosis with intrathecal trastuzumab administration is shown through this case report.

International audience; BACKGROUND: Several expression array studies identified molecular apocrine breast cancer (BC) as a subtype that expresses androgen receptor (AR) but not estrogen receptor alpha. We carried out a multicentre single-arm phase II trial in women with AR-positive, estrogen, progesterone receptor and HER2-negative (triple-negative) metastatic or inoperable locally advanced BC to assess the efficacy and safety of abiraterone acetate (AA) plus prednisone. PATIENTS AND METHODS: Patients with a metastatic or locally advanced, centrally reviewed, triple-negative and AR-positive (\textgreater/=10% by immunohistochemistry, IHC) BC were eligible. Any number of previous lines of chemotherapy was allowed. AA (1000 mg) was administered once a day with prednisone (5 mg) twice a day until disease progression or intolerance. The primary end point was clinical benefit rate (CBR) at 6 months defined as the proportion of patients presenting a complete response (CR), partial response (PR) or stable disease (SD) \textgreater/=6 months. Secondary end points were objective response rate (ORR), progression-free survival (PFS) and safety. RESULTS: One hundred and forty-six patients from 27 centres consented for IHC central review. Of the 138 patients with sufficient tissue available, 53 (37.6%) were AR-positive and triple-negative, and 34 of them were included from July 2013 to December 2014. Thirty patients were eligible and evaluable for the primary end point. The 6-month CBR was 20.0% [95% confidence interval (CI) 7.7%-38.6%], including 1 CR and 5 SD \textgreater/=6 months, 5 of them still being under treatment at the time of analysis (6.4+, 9.2+, 14.5+, 17.6+, 23.4+ months). The ORR was 6.7% (95% CI 0.8%-22.1%). The median PFS was 2.8 months (95% CI 1.7%-5.4%). Fatigue, hypertension, hypokalaemia and nausea were the most common drug-related adverse events; the majority of them being grade 1 or 2. CONCLUSIONS: AA plus prednisone treatment is beneficial for some patients with molecular apocrine tumours and five patients are still on treatment. CLINICALTRIALSGOV: NCT01842321

Background: Docetaxel and irinotecan chemotherapy have shown good efficacy in the treatment of advanced oesophago-gastric cancer. This randomised phase II study evaluated the efficacy and toxicity profile of two non-platinum docetaxel-based doublet regimens in advanced oesophago-gastric cancer. Methods: Chemotherapy-naïve patients with advanced oesophago-gastric cancer were randomised to receive either 3-weekly DI (docetaxel 60 mg m−2 plus irinotecan 250 mg m−2 (Day 1)) or 3-weekly DF (docetaxel 85 mg m−2 (Day 1) followed by 5-fluorouracil 750 mg m−2 per day as a continuous infusion (Days 1–5)). Results: A total of 85 patients received DI (n=42) or DF (n=43). The primary endpoint was overall response rate (ORR). The ORR and time to progression (TTP) in the evaluable population (n=65) were 37.5% (DI) vs 33.3% (DF), and 4.2 months vs 4.4 months, respectively. In the intent-to-treat population, the observed ORR, TTP and median overall survival were similar between the two groups. Grade 3–4 neutropenia, febrile neutropenia and diarrhoea were more frequent in the DI arm as compared with the DF arm (83.3% vs 69.8%, 40.5% vs 18.6%, and 42.9% vs 16.3%, respectively). Conclusion: Both docetaxel-based doublet regimens show comparable efficacy; however, the DF regimen was associated with a better toxicity profile and is an alternative treatment option for patients in whom platinum-based regimens are unsuitable.

Introduction Nous rapportons le premier cas d’erytheme pigmente fixe (EPF) au fulvestrant, confirme par patch-tests. Le fulvestrant est un agoniste des recepteurs aux œstrogenes, indique en premiere ligne chez les patients atteints de cancer du sein metastatique hormonodependant. Observation Une femme de 69 ans etait traitee par injections sous-cutanees de fulvestrant et denosumab (anticorps monoclonal diminuant la resorption osseuse) tous les 15 jours en alternance dans le cadre d’un cancer du sein metastatique. Apres quatre mois de traitement, elle presentait des lesions erythemateuses et prurigineuses bien delimitees du tronc, avec pigmentation sequellaire, evocatrices d’EPF. Les lesions recidivaient regulierement, evoquant la responsabilite du fulvestrant ou du denosumab. Il n’y avait pas d’autre medicament recemment introduit. La remission des lesions etait observee apres arret des deux medicaments. Les patch-tests au fulvestrant a la concentration de 250 mg/mL etaient positifs sur peau saine (dos) et sur sequelle pigmentee (region lombaire) a 48 h et 96 h. Les patch-tests au denosumab a la concentration de 70 mg/mL etaient negatifs sur les deux sites. L’examen histologique du patch-test positif au fulvestrant sur sequelle pigmentee mettait en evidence un infiltrat lichenoide avec necroses keratinocytaires confluentes, evoquant une reactivation de l’EPF sur le site du patch-test. L’immunohistochimie trouvait un infiltrat lympocytaire CD3+ significatif. Le patch-test au fulvestrant a la meme concentration effectue sur un volontaire sain consentant etait negatif. Aucune recidive des lesions n’etait constatee apres reprise du denosumab et remplacement du fulvestrant par l’exemestane (inhibiteur de la synthese des œstrogenes). Le diagnostic d’EPF au fulvestrant etait retenu. Une declaration en pharmacovigilance etait effectuee ( Fig. 1 et 2 ). Discussion Il s’agit a notre connaissance du premier cas decrit d’erytheme pigmente fixe au fulvestrant. Dans la base nationale de pharmacovigilance, seuls quatre cas de reactions cutanees sous fulvestrant seul ont ete rapportes : un cas de lesion nodulaire, deux cas d’erytheme au point d’injection et un cas d’eruption de type erytheme polymorphe. Aucun cas d’EPF n’est rapporte. Le diagnostic etiologique d’EPF est souvent realise sur un test de provocation positif, les tests epicutanes etant generalement peu sensibles. Dans notre cas, l’aspect histologique et immunohistochimique des lesions induites par les patch-tests au fulvestrant, la negativite du patch-test sur volontaire sain et l’evolution favorable des lesions apres arret du fulvestrant eliminent la possibilite de faux positifs. Conclusion L’EPF est une forme peu frequente de toxidermie. Il s’agit du premier cas decrit d’EPF au fulvestrant, confirme par explorations allergologiques et analyse histologique.

The risk of relapse after local therapy remains unacceptable for the majority of patients with early stage breast cancer. Adjuvant chemotherapy has been shown to prolong both disease-free survival (DFS) and overall survival. Several large phase III trials addressing the benefit of adding a taxane to conventional anthracycline-based regimen in early breast cancer showed a significant improves DFS in node-positive breast cancer patients. Taxane-based regimen seems to be more toxic in terms of myelosuppression. There seem to be fewer cardiac events after the taxane-based regimen attributable to the lower anthracycline cumulative dose. Therefore, the optimal adjuvant treatment in node positive breast cancer is still debated and several attempts are made to identify the patients, who will benefit most of taxane. The primary end point of this phase III randomized trial was to compare chemotherapy with 6 cycles of FEC100 versus 4 cycles of FEC100 followed by 4 cycles of Taxol, in patients with non-metastatic breast cancer with lymph node invasion (N+), in terms of DFS assessed by Kaplan-Meier analysis. The secondary end-points were overall survival (OS), local recurrence free interval, metastases free interval and tolerance. Material and Methods Between 10.3.2000 and 31.12.2002, 837 patients with invasive non-metastatic N+ breast cancer were randomized between (arm A) epirubicine 100 mg/m2 plus 5FU 500 mg/m2 plus cyclophosphamide 500 mg/m2 every 3 weeks (w) (FEC) for 6 cycles (417 patients) or (arm B) FEC every 3 w for 4 cycles then taxol 175 mg/m2 every 3 w for 4 cycles (FEC-T) (420 patients). Hormone-receptor-positive patients received either tamoxifen or anti-aromatize depending of their menopausal status, for 5 years after chemotherapy. Results The patients’ characteristics were well balanced between the 2 arms. The mean number of N+ was 4.5. Seventy-three % of the patients had oestrogen receptor positive (RO+) and 62 % progesterone receptor positive (RP+). The hazard ratio for DFS was 0.986 (95% confidence limits 0.773 — 1.257, P = 0.91). Five-year DFS rates were 78.5% with FEC and 78.4% with FEC-T. Nine-year DFS rates were 62.9% with FEC and 62.5% with FEC-T. The hazard ratio for OS was 0.848 (95% confidence limits 0.622 — 1.155, P = 0.29). The 5-y-OS was 86.1 % with FEC and 88.6 % with FEC-T) and the 9-y-OS was 73.9 % and 77 % respectively. The incidence of grade 3 to 4 neutropenia, infection, and nausea were higher with FEC without reaching significativity, neurotoxicity and hypersensitivity reactions were more frequent with FEC-T. There was no statistical difference in cardiac toxicity and febrile neutropenia between the 2 arms. Conclusion The final results of this trial suggest that adding taxol (every 3 weeks regimen) to an anthracycline-based regimen in patients with non-metastatic breast cancer with lymph node invasion (N+), confers little advantage in terms of DFS and OS. However, the present trial was powered to detect a 30% reduction in the risk of recurrence or death. A meta-analysis of all similar trials is needed to rule out smaller differences that could still be clinically worthwhile. This trial was supported by BMS Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-10-05.

Resume Les anticorps monoclonaux font de plus en plus partie de l’arsenal diagnostique et therapeutique. Les premieres utilisations in vivo ont ete dans les immunoscintigraphies pour lesquelles l’injection, par voie intraveineuse, d’anticorps monoclonal radiomarque permet la detection de tissu tumoral exprimant un antigene reconnu par cet anticorps. Il a ete decrit que ces anticorps pouvaient induire la production d’anticorps humains anti-souris (HAMA) chez ces patients. La presence de ces HAMA pourrait stimuler le systeme immunitaire avec un benefice sur la survie. Le cas presente ici est celui d’une patiente atteinte d’un adenocarcinome de l’ovaire decouvert a un stade metastatique. La patiente a recu dans un but diagnostique plusieurs injections d’anticorps monoclonaux et dans un but therapeutique un anticorps humanise. Les injections repetees des differents anticorps ont entraine une production importante de HAMA avec une augmentation des taux apres chaque reinjection. Le suivi biologique de la patiente a ete realise par le dosage de l’antigene CA125 et le dosage d’anticorps humains anti-souris (HAMA). Les taux eleves de HAMA chez cette patiente ont ete bien toleres et ont sans doute contribue a prolonger la survie de la patiente.

A revised kinematic model is proposed for the Neogene tectono-magmatic development of the North Tanzanian Divergence where the axial valley in S Kenya splits southwards into a wide diverging pattern of block faulting in association with the disappearance of volcanism. Propagation of rifting along the S Kenya proto-rift during the last 8 Ma is first assumed to have operated by linkage of discrete magmatic cells as far S as the Ngorongoro–Kilimanjaro transverse volcanic belt that follows the margin of cratonic blocks in N Tanzania. Strain is believed to have nucleated throughout the thermally-weakened lithosphere in the transverse volcanic belt that might have later linked the S Kenya and N Tanzania rift segments with marked structural changes along-strike. The North Tanzanian Divergence is now regarded as a two-armed rift pattern involving: (1) a wide domain of tilted fault blocks to the W (Mbulu) that encompasses the Eyasi and Manyara fault systems, in direct continuation with the Natron northern trough. The reactivation of basement fabrics in the cold and intact Precambrian lithosphere in the Mbulu domain resulted in an oblique rift pattern that contrasts with the orthogonal extension that prevailed in the Magadi–Natron trough above a more attenuated lithosphere. (2) To the E, the Pangani horst-like range is thought to be a younger (< 1 Ma) structure that formed in response to the relocation of extension S of the Kilimanjaro magmatic center. A significant contrast in the mechanical behaviour of the stretched lithosphere in the North Tanzanian diverging rift is assumed to have occurred on both sides of the Masai cratonic block with a mid-crustal decoupling level to the W where asymmetrical fault-basin patterns are dominant (Magadi–Natron and Mbulu), whereas a component of dynamical uplift is suspected to have caused the topographic elevation of the Pangani range in relation with possible far-travelled mantle melts produced at depth further N.

Resume Dix a 15 % des cancers du sein metastatiques developperont des metastases cerebrales symptomatiques. L'introduction du trastuzumab (Herceptin®) a ameliore les taux de reponse et la survie des patientes atteintes d'un cancer du sein metastatique surexprimant HER2+. Malgre la limite des etudes, retrospectives, d'effectif faible, de prise en charge differente, plusieurs auteurs soulignent une incidence de 30 % de metastases cerebrales chez les patientes avec un cancer du sein metastatique surexprimant HER2, traitees par trastuzumab alors que dans 70 a 80 % des cas la maladie est controlee en systemique. Dans le but d'un meilleur controle de la maladie au niveau central, un depistage en routine des metastases cerebrales chez des patientes a haut risque pourrait etre propose. L'apport du dosage des marqueurs tumoraux en tant qu'aide au diagnostic des metastases centrales pourrait etre interessant notamment quand la cytologie est negative, ce qui represente 30 % des cas, en raison d'une meilleure sensibilite de detection du processus tumoral par les dosages de marqueurs tumoraux.

At the time of hospitalization, it is essential to evaluate the general health status of a patient and to follow up the trends during therapy. Our work is focused on a set of tools for the measurement of patient activity. In this paper, we propose a few indicators of the patient activities of daily living, such as mobility, agitation, repartitions of stays, and displacements. As a result of this work, a diagnostic system was developed that could lead to a deeper knowledge of human activity rhythms in normal situations.

Resume Le cancer du sein est caracterise par son extreme frequence realisant un veritable probleme de sante publique. Une des composantes des processus de cancerisation est le dysfonctionnement des voies de communication cellulaires dont celle empruntant le recepteur Her-2. Il peut etre surexprime dans 30 % des cancers du sein et s'avere etre un facteur de mauvais pronostic, par une diminution de la survie globale et du delai de rechute. L'oncogene Her2/neu intervient dans la cancerogenese mammaire par amplification et/ou surexpression de son produit, la proteine Her-2 qui prend une importance considerable en tant que cible therapeutique. De nombreuses etudes demontrent l'interet d'utiliser le taux de Her-2 circulant comme outil de suivi therapeutique dans le cancer du sein metastatique surexprimant Her-2.

Resume Objectif de l'etude. – L'objectif de cette etude prospective, multicentrique et randomisee de phase III etait de comparer l'association de la fotemustine et d'une irradiation encephalique totale a la fotemustine seule en termes de reponse cerebrale et de duree de survie sans progression cerebrale chez des patients atteints de metastases cerebrales de melanome. Patients et Methodes. – Soixante-seize patients, au lieu des 106 patients prevus car l'essai a ete interrompu apres une analyse intermediaire, ont ete randomises pour recevoir la fotemustine seule (groupe A, n = 39) ou la fotemustine associee a une irradiation encephalique (groupe B, n = 37). La fotemustine a ete administree par voie intraveineuse a la dose de 100 mg m –2 aux jours 1, 8 et 15. Apres un repos therapeutique de 5 semaines, les patients dont le cancer n'a pas progresse ont recu toutes les 3 semaines 100 mg m –2 de fotemustine. Dans le groupe B, une irradiation encephalique a ete realisee concomitamment a la dose totale de 37,5 Gy en 15 fractions de 2,5 Gy et 3 semaines consecutives. Resultats. – Malgre l'existence de facteurs pronostiques plus defavorables chez les patients traites par la fotemustine seule, aucune difference significative n'a ete observee en termes de taux de reponse cerebrale objective (groupe A : 7,4 %, B : 10,0 %), de controle cerebral (reponses objectives et stabilisations) (groupe A : 30 %, B : 47 %) apres 7 semaines et de survie globale (groupe A : 86 j, B : 105 j). La duree de survie sans progression cerebrale etait significativement plus longue dans le groupe B ( p = 0,028, test de Wilcoxon). Conclusion. – L'association de la fotemustine et d'une irradiation encephalique a retarde la progression cerebrale par rapport a la fotemustine seule, mais n'a pas significativement ameliore le taux de controle objectif ou de survie globale chez des patients atteints de metastases cerebrales de melanome.

Since the publication of the Gynecologic Oncology Group (GOG) protocol 111 in 1996, and the results of the Arbeitgemeinschaft Gyna kologische Onkologie (AGO) trial Ovar-3 and the GOG protocol 158, the combination of platinum and paclitaxel has been adopted as the standard therapy in advanced ovarian cancer. One option for achieving further progress in the first-line treatment of advanced ovarian cancer might be the addition of noncross-resistant drugs to the two-drug regimen. Meta-analysis showed a survival benefit for platinum-anthracycline based combinations as compared to platinum-based combinations without anthracyclines. An AGO phase I/II trial compared epirubicin in combination with carboplatin and paclitaxel in untreated patients with gynecological malignancies. Based on the results of this study a randomized phase III trial together with the French GINECO group was conducted. The trial started 11/97 and was closed 11/99. All 1281 patients were randomized. Currently, 1132 end-of-therapy reports have been issued. Nine hundred eighty nine (87%) patients completed six cycles of treatment. Treatment and toxicity data are available for these patients. Three hundred thirty five patients had a measurable residual tumor after initial debulking surgery. Response data of 228 patients (111 ET-Carbo, 117 Carbo-T) are available.

Background: The balance between cell proliferation and drug-induced cell death by apoptosis or necrosis plays a major role in determining response to chemotherapy. Commonly-used DNA analysis methods cannot study both parameters simultaneously. A new approach described here combines a green fluorescent membrane-intercalating dye (PKH67) with Hoechst 33342 or annexin V and propidium iodide, to allow simultaneous assessment of cell division, cell cycle status, apoptosis, and necrosis, respectively. Methods: To test this approach, we used cultured K562 leukemic cell lines which are drug-sensitive (K562S) or drug-resistant (K562R) by virtue of whether they lack or exhibit expression, respectively, of the gp-170 (PGP) glycoprotein pump involved in multidrug resistance. Results: We found that: 1) PKH67 fluorescence intensity decreases proportionately to number of cell divisions, 2) labeling with PKH67 does not alter either cell cycle distribution, as assessed by vital DNA staining with Hoechst 33342, or cell growth, and 3) using a simple threshold analysis method suitable for real-time sorting decisions, subpopulations of proliferating cells present at initial levels of ≥ 10% can readily be detected after two cell division times, based on decreased PKH67 intensity. Finally, we demonstrated that after treatment of an admixture of K562S and K562R with vincristine, triple-labeling with PKH67, annexin V, and propidium iodide can be used to identify and sort those cells which remain not only viable (nonnecrotic, nonapoptotic) but actively dividing (decreased PKH67 intensity) in the presence of drug. Conclusions: Although the studies described here were carried out in a model system using cells having known drug resistance phenotypes, we expect that the methods described will be useful in ex vivo studies of clinical leukemic specimens designed to identify the role played by specific chemoresistance proteins and mechanisms in therapeutic outcomes for individual patients.Cytometry (Comm. Clin. Cytometry) 42:50–60, 2000 © 2000 Wiley-Liss, Inc.

Despite the generalization of induction chemotherapy and a better outcome for chemosensitive diseases, the prognosis of inflammatory breast cancer (IBC) is still poor. In this work, we evaluate response and toxicity of high-dose sequential chemotherapy with repeated blood stem cell (BSC) transplantation administered as initial treatment in 100 women with non-metastatic IBC. Ninety-five patients (five patients were evaluated as non-eligible) of median age 46 years (range 26–56) received four cycles of chemotherapy associating: cyclophosphamide (C) 6 g m−2 – doxorubicin (D) 75 mg m−2 cycle 1, C: 3 g m−2 – D: 75 mg m−2 cycle 2, C: 3 g m−2 – D: 75 mg m−2 – 5 FU 2500 mg m−2 cycle 3 and 4. BSC were collected after cycle 1 or 2 and reinfused after cycle 3 and 4. rG-CSF was administered after the four cycles. Mastectomy and radiotherapy were planned after chemotherapy completion. Pathological response was considered as the first end point of this trial. A total of 366 cycles of chemotherapy were administered. Eighty-seven patients completed the four cycles and relative dose intensity was respectively 0.97 (range 0.4–1.04) and 0.96 (range 0.25–1.05) for C and D. Main toxicity was haematological with febrile neutropenia ranging from 26% to 51% of cycles; one death occurred during aplasia. Clinical response rate was 90% ± 6%. Eighty-six patients underwent mastectomy in a median of 3.5 months (range 3–9) after the first cycle of chemotherapy; pathological complete response rate in breast was 32% ± 10%. All patients were eligible to receive additional radiotherapy. High-dose chemotherapy with repeated BSC transplantation is feasible with acceptable toxicity in IBC. Pathological response rate is encouraging but has to be confirmed by final outcome. © 1999 Cancer Research Campaign

Summary Background Platinum compounds are the most active drugs in ovarian cancer treatment; cisplatin and carboplatin demonstrated similar efficacies but different toxicity profiles. Paclitaxel combined with cisplatin as first-line treatment improved overall survival when compared to a cisplatin–cyclophosphamide combination, but generated higher rates of neutropenia, febrile neutropenia and neurotoxicity. The paclitaxel–carboplatin combination may be better tolerated than cisplatin–paclitaxel. Design The objective of the present study was to assess the efficacy and safety of the combination of paclitaxel and carboplatin in previously treated advanced ovarian cancer patients. Patients and methods During or after platinum-based chemotherapy, 73 patients with progressive advanced epithelial ovarian carcinoma were enrolled to receive every four weeks a three-hour infusion of paclitaxel 175 mg/m2 followed by a 30-minute carboplatin infusion. The carboplatin dose was calculated to obtain the recommended area concentration versus-time under the curve of 5 mg·ml− 1·min. Results Toxicity and response could be evaluated for 72 and 62 patients, respectively. Eleven complete and 15 partial responses gave an overall response rate of 42% (95% CI: 30%–54%). Response rates for platinum-refractory patients and those with early (≥ 3 and 12 months) relapses were 24%, 33% and 70%, respectively. The respective median response duration, the median progression-free survival and median overall survival were 8, 6 and 14 months. Myelosuppression was the most frequent and severe toxicity. Grade 3 and 4 neutropenia occurred, respectively in 30% and 23% of the cycles; 6% of the cycles benefited from medullary growth factors. Only one episode of febrile neutropenia was observed. Grade 3 and 4 thrombocytopenia occurred, respectively during 3% and 1% of the cycles. Alopecia was frequent. Transient peripheral neuropathy developed in 47% of patients but was severe in only one patient. One early death was attributed to progressive disease and possibly to therapy. Conclusion This combined paclitaxel–carboplatin therapy is effective and can be safely administered to ovarian cancer patients who relapse after one or two regimens of platinum-based chemotherapy.

The wide discrepancies in the frequency of ‘positive’ samples for multidrug resistance (MDR) phenotype within the same type of tumor observed in the literature justified the need for the definition of consensus recommendations. To define standard techniques of MDR phenotype measurement, we ran a large multicentric evaluation of the different methods available. Thirty-six French centers participated in the study, and 742 samples of 2–10 × 106 viable cells were sent by overnight express mail between December 1993 and February 1996. The same batches of MRK16, 4E3 and UIC2 were used. Nineteen samples of leukemia (12 AML, 1 ALL, 6 lymphoproliferative syndromes) and six leukemic cell lines with different levels of MDR expression were tested. Five meetings reached agreement concerning the guidelines for each technique, except immunocytochemistry. The 19 fresh samples were tested by each center using one to four techniques among cytofluorometry, immunocytochemistry, functional tests and RT-PCR. Five samples were diagnosed as ‘negative’ according to local criteria, with few discordant results (0 to 16% of ‘positive’ results). For all the 14 remaining samples, large discrepancies were observed from center to center, and from one technique to another. No correlations could be found between techniques. Flow cytometric analysis of cells already exposed to MRK16 or control IgG2A, fixed in paraformaldehyde and sent to centers did not reduce the discrepancies between centers in two of the four samples with moderate expression, emphasizing the role of histogram interpretation. The use of alternative monoclonal antibodies (4E3 and UIC2) did not reduce the discrepancies observed. In a second step, the K562 parental cell line, a low resistant subline (K562/HHT100, ×7 resistance index to DNR) and a high resistant subline (K562/HHT300, ×125 resistance index to DNR) were sent blindly three times, with an increasing level of recommendations for flow cytometry. Dramatic improvements were observed in cytometric results when the result was expressed as the ratio of arithmetic mean of fluorescence of antibody (10 μg of MRK16)/arithmetic mean of fluorescence of control (10 μ g IgG2A): the proportion of expected results increased from 61 to 100% for K562, and from 37 to 85% for K562/HHT100. For uptake and drug efflux measurements, the use of 1 h uptake of 0.1μ M of rhodamine, followed by 1 h efflux ±10 μ M of verapamil, permitted an increased reproducibility of the technique from 71 to 100% for K562 and K562/HHT100. Whatever the technique used, concordant results were obtained for K562/HHT300. The immunocytochemistry, using several antibodies (MRK16, JSB1 and C219) gave many non-interpretable results (44%), due to a frequent high background and discordant results between antibodies in the same centers, and discordant conclusions between centers. The group does not recommend this technique for circulating tumoral cells.

Since there is no consensus on the techniques for multidrug resistance (MDR) phenotype evaluation, many discrepancies concerning the importance and frequency of mdr1 gene expression in leukemias and solid tumors are observed in the literature. In order to establish an inter-laboratory consensus in France, a multicenter study was carried out to propose further guidelines for MDR phenotype evaluation. The techniques used by the 38 laboratories participating in the trial were: immunodetection (immunohisto and/or cytochemistry, flow cytometry), functional tests, reverse transcription-polymerase chain reaction (RT-PCR) or Northern blot. We present the results obtained by 19 laboratories concerning the measurement of mdr1 gene expression assessed by RT-PCR or Northern blot in: (1) 19 samples of tumor cells obtained from leukemic patients; (2) six solid tumor samples obtained at surgery; (3) eight cell lines exhibiting variable levels of resistance, and; (4) 10 preparations of RNA and of cDNA obtained from solid tumors. Standardization of the RT-PCR technique and preliminary results comparing RT-PCR with immunohistochemistry in solid tumors are also reported.

PURPOSE The purpose of this study was to evaluate the efficacy and tolerance of recombinant human interferon gamma (rIFN-gamma) as second-line treatment in patients with persistent disease at second-look laparotomy. PATIENTS AND METHODS One hundred eight patients with residual disease at second-look laparotomy were treated with rIFN-gamma (20 x 10(6) IU/m2) administered intraperitoneally (IP) twice a week for 3 to 4 months. In the absence of clinically assessable disease, response to rIFN-gamma was assessed with a third-look laparotomy. RESULTS Of 98 assessable patients, 31 (32%) achieved a surgically documented response, including 23 patients (23%) with a complete response (CR). The age and size of residual tumor were significant prognostic factors for the response to rIFN-gamma. A 41% CR rate was observed in 41 patients younger than 60 years and with residual tumor less than 2 cm. The probability of response was independent of previous response to first-line chemotherapy. The median duration of response was 20 months and the 3-year survival rate in responders was 62%. Response to rIFN-gamma was the most significant prognostic factor for survival of patients with residual disease. Adverse events included fever, flu-like syndrome, neutropenia, and liver enzyme disturbances. No significant peritoneal fibrosis was noted. CONCLUSION These results support the potential interest of IP rIFN-gamma as adjuvant treatment in ovarian cancer. Controlled prospective trials are required to determine its place in the therapeutic strategy of this malignancy.

Le trastuzumab (Herceptin®) est un composant cle du traitement standard des patientes (pts) avec cancer du sein (CS) precoce HER2 positif [1] et est autorise en administration intraveineuse (IV). La dose de charge initiale (8 mg/kg) est administree en IV en 90 minutes, suivie par des doses d’entretien de 6 mg/kg/3 sem en 30 minutes. Une nouvelle formulation sous-cutanee (SC) de trastuzumab, avec une dose fixe de 600 mg et pour excipient une hyaluronidase recombinante humaine (rHuPH20), a ete developpee comme alternative au traitement IV. Cette formulation SC permet une administration rapide (< 5 minutes) ameliorant potentiellement la praticabilite et la compliance du traitement.

To evaluate the feasibility and the efficacy of an intensive alternating chemotherapy regimen with hematopoietic growth factor support in the late salvage treatment of patients with metastatic nonseminomatous germ cell tumors (NSGCT), 14 patients with refractory or recurrent disease were treated with a combination regimen of vinblastine and bleomycin (VB) followed by three weekly cycles of BOP (bleomycin + vincristine + cisplatin) and subsequent CISCA (cyclophosphamide + doxorubicin + cisplatin) cycles. All patients experienced a grade 3 or 4 neutropenia despite prophylactic growth factor support; II patients required empiric antimicrobial therapy during the chemotherapy program. No toxic death or other significant adverse effect was seen. Eight patients achieved a complete remission and three patients had a partial remission with normalization of serum tumor markers at completion of chemotherapy. Four patients received high-dose chemotherapy with autologous bone marrow transplantation as consolidation therapy and one patient underwent a salvage surgery. Three patients remain free of disease at over 18, 26, and 36 months after therapy. We demonstrate here the feasibility of an intensive alternating chemotherapy schedule in the late salvage treatment of progressive NSGCT. Our results also suggest that the combination of such an approach with high-dose chemotherapy (HDCT) and/or surgery can be used in a curative attempt after the failure of first line salvage therapy.

The effects of elevated atmospheric CO(2) (700 micro mol mol(-1)) and fertilization were investigated on 2-year-old sweet chestnut (Castanea sativa Mill.) seedlings grown outdoors in pots in constantly ventilated open-sided chambers. Plants were divided into four groups: fertilized controls (+F/-CO(2)), unfertilized controls (-F/-CO(2)), fertilized + CO(2)-treated plants (+F/+CO(2)) and unfertilized + CO(2)-treated plants (-F/+CO(2)). Dry matter accumulation and allocation were measured after one growing season and CO(2) exchange of whole shoots was measured throughout the growing season. Shoot growth and total leaf area of unfertilized plants were not affected by elevated CO(2), whereas both parameters were enhanced by elevated CO(2) in fertilized plants. Elevated CO(2) increased total biomass by about 20% in both fertilized and unfertilized plants; however, biomass partitioning differed. In unfertilized plants, elevated CO(2) caused an increase in root growth, whereas in fertilized plants, it stimulated aboveground growth. At the whole-shoot and leaf levels, photosynthetic activity of both fertilized and unfertilized plants increased in response to elevated CO(2), but the seasonal pattern of this enhancement varied with nutrient treatment. In unfertilized plants, a downward acclimation of photosynthesis was observed early in the season (June), and was related to reductions in nitrogen and chlorophyll content and to starch accumulation. The decrease in the slope of the A/C(i) curve suggested a decrease in Rubisco activity. In both fertilized and unfertilized plants, shoot respiration decreased during the night in response to elevated CO(2) until mid-July. The decrease was not related to changes in sugar concentration.

Two-year-old sweet chestnut trees were grown outside in normal or double CO2 atmospheric concentration. In spring and in autumn of two growing seasons, a six day labelling pulse of14C labelled CO2 was used to follow the carbon assimilation and distribution in the plant-soil system. Doubling atmospheric CO2 had a significant effect on the tree net carbon uptake. A large proportion of the additional C uptake was ‘lost’ through the root system. This suggests that increased C uptake under elevated CO2 conditions increases C cycling without necessarily increasing C storage in the plant. Total root derived material represented a significant amount of the ‘extra-assimilated’ carbon due to the CO2 treatment and was strongly correlated with the phenological stage of the tree. Increasing root rhizodeposition led to a stimulation of microbial activity, particularly near the end of the growing season. When plant rhizodeposition was expressed as a function of the root dry weight, the effect of increasing CO2 resulted in a higher root activity. The C to N ratios were significantly higher for trees grown under elevated CO2 except for the fine root compartment. An evaluation of the plant-soil system nitrogen dynamics showed, during the second season of CO2 treatment, a decrease of soil N mineralization rate and total N uptake for trees grown at elevated CO2 levels.

Resume L'apparition de HAMA ( human anti-mouse antibodies ) par allo-immunisation apres injection d'un anticorps murin radiomarque est le principal inconvenient de l'immunoscintigraphie. La presence de HAMA peut notamment perturber les immunodosages et entrainer des resultats errones dont il est absolument necessaire de s'affranchir. Notre experience a propos de cinq patientes ayant une telle alloimmunisation nous amene a conclure que le traitement systematique du serum par chromatographie sur proteine A est actuellement le moyen le plus fiable, a la fois pour detecter les HAMA et se debarrasser de l'interference.

We investigated four mechanisms of intrinsic chemoresistance in a series of 67 human brain tumours including 31 gliomas (one grade I ganglioglioma, nine grade II and 10 grade III astrocytomas, 11 glioblastomas), 13 cerebral metastases, one medulloblastoma, one malignant teratoma, three ependymomas and 18 meningiomas. We studied four genes by northern blotting: multidrug-resistance (MDR 1), glutathione-s transferase (GST pi), dihydrofolate reductase (DHFR), and topoisomerase II (Topo II). The Topo II gene was absent in the normal adult brain (100%) and in 64% of the tumour samples tested. A second gene, GST pi, was found to be overexpressed in 38% of brain tumours. The two other chemoresistance-related genes were occasionally overexpressed in brain tumours (2% for MDR1, 9% for DHFR). Our results provide evidence that chemoresistance is intrinsic to the brain tissue and seems likely to be a multifactorial process.

C02 enrichment of the atmosphere is now well documented and its effect on the growth of world forests is being questioned by the scientific community. The direct effects of increased C02 on tree species are reviewed: the different experimental approaches are described, as well as the principal results already obtained. Short-term experiments have shown an increased photosynthetic rate, as predicted by leaf models. In longer experiments this increase is reduced after a few weeks or months by mechanisms that remain to be found. Elevated C02 seems to decrease the dark respiration rate, but the results are still controversial. Biomass partitioning in elevated C02 is clearly related to the mineral supply of the trees: An increase in root investment in elevated C02 is related to a poor mineral status. The mineral content of trees grown in elevated C02 is generally lowered compared to controls. No general rule has yet been found for the effect of increased C02 on leaf area development. The paper emphasizes large areas of ignorance: the reasons for the different responses of different species, which may be related to their developmental strategies, are largely ignored. Much experimental effort is needed to parameterize all the physiological processes which are susceptible to change with an increase in atmospheric C02, leading to a change in forest tree growth.

On a etudie l'effet d'un doublement de la concentration en CO 2 de l'atmosphere (soit 350 vpm, teneur actuelle et 700 vpm) sur la repartition de la biomasse et du contenu en azote chez de jeunes plants de châtaigniers (Castanea sativa Mill). Les arbres, âges de 2 ans, sont cultives en pots a l'exterieur pendant toute une saison de vegetation sous des tunnels ou miniserres recouvertes de propafilm et ventilees en permanence. Le doublement du CO 2 ambiant est obtenu par addition constante de CO 2 pur d'origine industrielle. Ces jeunes châtaigniers sont cultives sous nutrition minerale contrastee (sol forestier auquel est ajoute ou non de l'engrais NPK en granules)(.)

The dynamic interpretation of the serum concentrations of markers is frequently used to calculate biological indicators of therapeutic efficiency and relapse. But analysis of marker kinetics can also help improve our understanding of the natural history of different types of cancer and their evolution under treatment. This application is illustrated by an analysis of CA 125 kinetics measured in a patient with stage III ovarian cancer treated with multiple lines of chemotherapy and who had a survival time of 9 years.

LV5FU2 with high-dose leucovorin (LV), weekly infusional 5-fluorouracil (5FU) (AIO schedule) and raltitrexed have been demonstrated to be active agents in first-line treatment of colorectal cancer. We performed a 4-arm randomised trial to compare (1) a low-dose intravenous bolus of LV (20 mg/m2), followed by an intravenous bolus of 5FU (400 mg/m2), followed by a 22-hour continuous infusion of 5FU (600 mg/m2) on day 1 and day 2/2 weeks (ldLV5FU2 arm), (2) a weekly continuous infusion of high-dose 5FU (2.6 g/m2/week) for 6 weeks followed by a rest week (HD-FU arm) and (3) raltitrexed (Tomudex arm; 3 mg/m2/3 weeks) to standard LV5FU2. From 1997 to 2001, 294 patients were included. The 4 arms were well balanced for sex ratio, age, WHO performance status, the primary tumour site and prior adjuvant chemotherapy. Treatment was stopped due to low accrual. Two toxicity-related deaths were observed in the Tomudex arm. The treatments gave rise to different rates of grade 3-4 neutropenia (3, 4, 11 and 14% of the patients in the LV5FU2, ldLV5FU2, HD-FU and Tomudex arms, respectively, p = 0.028), leucopenia and vomiting. At least one episode of grade 3-4 toxicity was observed in 27, 25, 38 and 47% of the patients in the LV5FU2, ldLV5FU2, HD-FU and Tomudex arms, respectively (p = 0.016). An objective response was observed in 28, 21, 22 and 10% of the patients in the LV5FU2, ldLV5FU2, HD-FU and Tomudex arms, respectively (p = 0.04). Progression-free survival (PFS) of the patients in the Tomudex arm was statistically lower compared to that of patients treated with LV5FU2 or ldLV5FU2 (combined group; p = 0.013, log rank test). In conclusion, Tomudex is more toxic and yields shorter PFS than infusional 5FU. Despite the early closure of the study and the lack of power of the comparison, it seems that ldLV5FU2 could be considered as an active, easier and less expensive option for the treatment of metastatic colorectal cancer compared to classic LV5FU2 or weekly HD-FU.

SUMMARY A Mw 4.3 earthquake occurred on 2002 September 30, in the Armorican Massif, NW France. Since it was one of the largest events ever recorded in this region, this was the opportunity to improve our seismotectonic knowledge of the Armorican Massif. We performed a post-seismic survey (SISBREIZH), which allowed us to locate accurately 62 aftershocks within 14 days. An analysis of the main shock using broadband records provided a normal fault mechanism with a dextral strike-slip component located at 12-km depth. The aftershock sequence exhibits: (1) a combination of almost pure right-lateral strike-slip and dominant normal faulting similar to the main shock; (2) magnitudes ranging from 0.4 to 1.9 and (3) depths ranging from 11.5 to 13.5 km, that is, close to the main shock hypocenter. The distribution of the aftershocks defines a rupture plane dipping 60° to the south with a fault length of ≈2 km consistent with the source parameters of the main shock. Beside the SISBREIZH survey, a morpho-structural analysis has been conducted: we found fault plane solutions with southward-dipping N120–150 normal fault planes. The stress tensor computed after the aftershock focal mechanisms is a strike-slip regime with a NE–SW extensional direction. The Lorient earthquake appears to reactivate Late Hercynian structures and the whole sequence is reflecting the regional-scale tectonic stress field expressed by a combination of strike-slip and normal faulting.

The main objective of this prospective multicenter randomised phase III study was to compare a combined regimen of fotemustine plus whole brain irradiation versus fotemustine alone in terms of cerebral response and time to cerebral progression in patients with melanoma brain metastases.Seventy-six patients (instead of the 106 planned patients; study was stopped after the interim analysis) were randomised receiving either fotemustine (arm A, n = 39) or fotemustine and whole brain irradiation (arm B, n = 37). Fotemustine was administered intravenously at 100 mg m(-2) on day 1, 8 and 15, followed by a 5-week rest period, then every 3 weeks in non-progressive patients. In arm B, a concomitant whole brain irradiation was performed at the total dose of 37.5 Gy (2.5 Gy/d(-1), days 1-5, 3 consecutive weeks).Although patients who received fotemustine alone had worse prognostic factors, there was no significant difference in brain response (arm A: 7.4%, B: 10.0%) or control rates (objective response plus stable disease) after seven weeks (arm A: 30%, B: 47%) and overall survival (arm A: 86d, B: 105d). However, there was a significant difference in favour of arm B for the time to brain progression (p = 0.028, Wilcoxon test).Fotemustine plus whole brain irradiation delayed the time to brain progression of melanoma cerebral metastases compared to fotemustine alone but without a significant improvement in terms of objective control or overall survival.

The main objective of this prospective multicentre randomized phase III study was to compare a combined regimen of fotemustine plus whole brain irradiation with fotemustine alone in terms of cerebral response and time to cerebral progression in patients with melanoma cerebral metastases. Seventy-six patients were randomized to receive either fotemustine (arm A, n = 39) or fotemustine plus whole brain irradiation (arm B, n = 37). Fotemustine was administered intravenously at 100 mg/m(2) on days 1, 8 and 15, followed by a 5 week rest period, then every 3 weeks in non-progressive patients. In arm B, concomitant whole brain irradiation was performed at a total dose of 37.5 Gy (2.5 Gy/day on days 1-5 for three consecutive weeks). Although patients who received fotemustine alone had worse prognostic factors, there was no significant difference in cerebral response (arm A, 7.4%, arm B, 10.0%) or control rates (objective responses plus stable disease) after 7 weeks (arm A, 30%; arm B, 47%) or in overall survival (arm A, 86 days; arm B, 105 days). However, there was a significant difference in favour of arm B for the time to cerebral progression (P = 0.028, Wilcoxon test). In conclusion, fotemustine plus whole brain irradiation delayed the time to cerebral progression of melanoma cerebral metastases compared with fotemustine alone but without a significant improvement in terms of objective control or overall survival.

Proliferation and multidrug resistance status are key predictors of therapeutic outcome in acute myeloid leukaemia (AML). Anthracyclines such as daunorubicin (DNR) are typically used to treat AML and can induce drug resistance. The goal of the studies described here was to select a combination of fluorescent probes that could be used in combination with flow cytometry to monitor cell proliferation vs. cell death/necrosis as a function of anthracycline uptake. Propidium iodide (PI), the most commonly used marker of membrane integrity, cannot be used to evaluate necrosis in DNR‐containing cells because of spectral overlap. A membrane integrity probe compatible with the use of a dye dilution method using PKH67 to study cell proliferation was also selected. The results show that DAPI and Cascade Blue (CB), like PI, were able to detect necrotic cells when no DNR was present, although CB gave less resolution between viable and necrotic cells than PI or DAPI. In the presence of DNR, DAPI cannot be used owing to the fluorescence quenching by DNR. However, it was found that a combination of DNR, CB, and PKH67 allows simultaneous identification of chemoresistant cells, based on reduced DNR accumulation, necrotic cells based on CB incorporation, and proliferating cells based on partitioning of PKH67 fluorescence between daughter cells. It was also found that unless a marker of necrosis is used in combination with the dye dilution assay, a moderate decrease of fluorescence as a result of necrosis may be incorrectly interpreted as proliferation.

Since the publication of the Gynecologic Oncology Group (GOG) protocol 111 in 1996, and the results of the Arbeitgemeinschaft Gyna kologische Onkologie (AGO) trial Ovar-3 and the GOG protocol 158, the combination of platinum and paclitaxel has been adopted as the standard therapy in advanced ovarian cancer. One option for achieving further progress in the first-line treatment of advanced ovarian cancer might be the addition of noncross-resistant drugs to the two-drug regimen. Meta-analysis showed a survival benefit for platinum-anthracycline based combinations as compared to platinum-based combinations without anthracyclines. An AGO phase I/II trial compared epirubicin in combination with carboplatin and paclitaxel in untreated patients with gynecological malignancies. Based on the results of this study a randomized phase III trial together with the French GINECO group was conducted. The trial started 11/97 and was closed 11/99. All 1281 patients were randomized. Currently, 1132 end-of-therapy reports have been issued. Nine hundred eighty nine (87%) patients completed six cycles of treatment. Treatment and toxicity data are available for these patients. Three hundred thirty five patients had a measurable residual tumor after initial debulking surgery. Response data of 228 patients (111 ET-Carbo, 117 Carbo-T) are available.

Anthracyclin-based chemotherapy is the most efficient chemotherapy for advanced or metastatic soft tissue sarcoma (STS). Development of anthracyclin chemoresistance has been widely documented. In a previous clinical trial, we evaluated a possible reversal of anthracyclin chemoresistance after exposure to subcutaneous IL-2. The current phase II clinical study entered 17 proven metastatic STS patients, refractory to anthracyclin chemotherapy, who received IL-2, and subsequent anthracyclin-based chemotherapy. Subcutaneous IL-2 was administered at 18 million Units/day, 5 days a week for two consecutive weeks. Treatment was administered safely at the full dose for 16 out of 17 patients, and toxicity was mild. One patient had treatment stopped because of rapidly progressive disease. As soon as patients met biological and clinical criteria, chemotherapy was administered. The median delay was 12 days (2-23) from the end of IL-2 administration. Only 13 patients received anthracyclin chemotherapy after IL-2. The other 4 patients did not receive chemotherapy for progressive disease. One partial response was observed out of 13 evaluable patients (7.7% overall response, 95% confidence interval: 0.2 to 36). The overall response rate was 5.9% (95% CI: 0.15 to 29), so the study was stopped due to lack of efficacy. In previous and current studies, a few patients have developed restored anthracyclin chemosensitivity following exposure to IL-2. No conclusive evidence of IL-2 chemoresistance reversal was obtained from this study. Further investigations need to be performed with perhaps a larger group of more carefully selected patients using a different schedule and sequence of combined cytokines and chemotherapy.

The balance between cell proliferation and drug-induced cell death by apoptosis or necrosis plays a major role in determining response to chemotherapy. Commonly-used DNA analysis methods cannot study both parameters simultaneously. A new approach described here combines a green fluorescent membrane-intercalating dye (PKH67) with Hoechst 33342 or annexin V and propidium iodide, to allow simultaneous assessment of cell division, cell cycle status, apoptosis, and necrosis, respectively.To test this approach, we used cultured K562 leukemic cell lines which are drug-sensitive (K562S) or drug-resistant (K562R) by virtue of whether they lack or exhibit expression, respectively, of the gp-170 (PGP) glycoprotein pump involved in multidrug resistance.We found that: 1) PKH67 fluorescence intensity decreases proportionately to number of cell divisions, 2) labeling with PKH67 does not alter either cell cycle distribution, as assessed by vital DNA staining with Hoechst 33342, or cell growth, and 3) using a simple threshold analysis method suitable for real-time sorting decisions, subpopulations of proliferating cells present at initial levels of/= 10% can readily be detected after two cell division times, based on decreased PKH67 intensity. Finally, we demonstrated that after treatment of an admixture of K562S and K562R with vincristine, triple-labeling with PKH67, annexin V, and propidium iodide can be used to identify and sort those cells which remain not only viable (nonnecrotic, nonapoptotic) but actively dividing (decreased PKH67 intensity) in the presence of drug.Although the studies described here were carried out in a model system using cells having known drug resistance phenotypes, we expect that the methods described will be useful in ex vivo studies of clinical leukemic specimens designed to identify the role played by specific chemoresistance proteins and mechanisms in therapeutic outcomes for individual patients.