Potential target antigens for immunotherapy in human pancreatic cancer.

BACKGROUND: To be effective and selective, immunotherapy ideally targets specifically tumor cells and spares normal tissues. Identification of tumor specific antigens is a prerequisite to establish an effective immunotherapy. Still very little is known about the expression of tumor-related antigens in pancreatic neoplasms. Cancer Testis antigens (CT) are antigens shared by a variety of malignant tumors, but not by normal tissues with the exception of germ cells in testis. Restricted expression in neoplastic tissues and inherent immunogenic features make CT antigens ideal for use in immunotherapy. We analyzed the expression of a selected panel of nine CT antigens that have been proven to elicit an efficient immunogenic response in other malignancies. In addition we analyzed the expression of HERV-K-MEL, an immunogenic antigen of viral origin. METHODS: Pancreatic adenocarcinoma tumor samples (n=130) were obtained intraoperatively, control tissues (n=23) were collected from cadaveric donor and from patients with chronic pancreatitis. Tumor-associated antigen expression of MAGE-A1, MAGE-A3, MAGE-A4, MAGE-A10, LAGE-1, NY-ESO-1, SCP-1, SSX-2, SSX-4 and HERV-K-MEL was assessed by PCR. Sequencing of PCR products were performed to assess the expression of SSX-4 in neoplastic and normal pancreatic tissues. RESULTS: Three of 10 tested antigens were expressed in over 10% of malignant pancreatic tissue samples. SSX-4 was found positive in 30% of cases, SCP-1 in 19% and HERV-K-MEL in 23% of cases. No expression of CT antigens was found in non-malignant pancreatic tissue with the exception of SSX-4 and and SSX-2. CONCLUSIONS: Fifty two percentage of the analyzed tissues expressed at least one CT antigen. The concomitant expression of SSX-4 in both malignant and non-malignant pancreatic tissue is a new finding which may raise concerns for immunotherapy. However, HERV-K-MEL is expressed with a relatively high prevalence and may be a candidate for specific immunotherapy in a large subgroup of pancreatic cancer patients. This study advocates the analysis of patients with regard to their immunogenic profile before the onset of antigen-specific immunotherapy.

Sacramentarium Gelasianum, pars.

F. 1 Poenitentiale Cummeani, c. VI, 22-IX, 10 et c. XIII, II-XIV, 19 (Schmitz, Bussbücher, I, 631-636 et 642-645.) F. 4v « Incipit liber sacramentorum ». F. 5-8v et 16-27v Temporal, commençant à la vig. de Noël. F. 9v-14 Sanctoral. F. 14 Préface, Canon incompl., avec mentions de s. Hilaire et s. Martin (15v). F. 28 Oraisons « super populum ». F. 28v « Incipit liber secundus de extrema parte ». F. 30 Célébration du baptême ; « Ad succurrendum infirmum catecuminum » (Martène, De ant. eccl. rit., I, 182) (32) ; confirmation (33v). F. 34v « Ordo ad paenitentiam dandam » (ibid., 803). F. 35 Bénédictions, oraisons et messes diverses. F. 43v « Oratio ad missam pro pace » (incompl.).

Saturable metabolism of continuous high-dose ifosfamide with mesna and GM-CSF: a pharmacokinetic study in advanced sarcoma patients. Swiss Group for Clinical Cancer Research (SAKK).

BACKGROUND: The aim of this study was to assess the pharmacology, toxicity and activity of high-dose ifosfamide mesna +/- GM-CSF administered by a five-day continuous infusion at a total ifosfamide dose of 12-18 g/m2 in adult patients with advanced sarcomas. PATIENTS AND METHODS: Between January 1991 and October 1992 32 patients with advanced or metastatic sarcoma were entered the study. Twenty-seven patients were pretreated including twenty-three with prior ifosfamide at less than 8 g/m2 total dose/cycle. In 25 patients (27 cycles) extensive pharmacokinetic analyses were performed. RESULTS: The area under the plasma concentration-time curve (AUC) for ifosfamide increased linearly with dose while the AUC's of the metabolites measured in plasma by thin-layer chromatography did not increase with dose, particularly that of the active metabolite isophosphoramide mustard. Furthermore the AUC of the inactive carboxymetabolite did not increase with dose. Interpatient variability of pharmacokinetic parameters was high. Dose-limiting toxicity was myelosuppression at 18 g/m2 total dose with grade 4 neutropenia in five of six patients and grade 4 thrombocytopenia in four of six patients. Therefore the maximum tolerated dose was considered to be 18 g/m2 total dose. There was one CR and eleven PR in twenty-nine evaluable patients (overall response rate 41%). CONCLUSION: Both the activation and inactivation pathways of ifosfamide are non-linear and saturable at high-doses although the pharmacokinetics of the parent drug itself are dose linear. Ifosfamide doses greater than 14-16 g/m2 per cycle appear to result in a relative decrease of the active metabolite isophosphoramide mustard. These data suggest a dose-dependent saturation or even inhibition of ifosfamide metabolism by increasing high dose ifosfamide and suggest the need for further metabolic studies.

Neoadjuvant chemotherapy and radiotherapy followed by surgery in selected patients with stage IIIB non-small-cell lung cancer: a multicentre phase II trial.

BACKGROUND: Stage IIIB non-small-cell lung cancer (NSCLC) is usually thought to be unresectable, and is managed with chemotherapy with or without radiotherapy. However, selected patients might benefit from surgical resection after neoadjuvant chemotherapy and radiotherapy. The aim of this multicentre, phase II trial was to assess the efficacy and toxicity of a neoadjuvant chemotherapy and radiotherapy followed by surgery in patients with technically operable stage IIIB NSCLC. METHODS: Between September, 2001, and May, 2006, patients with pathologically proven and technically resectable stage IIIB NSCLC were sequentially treated with three cycles of neoadjuvant chemotherapy (cisplatin with docetaxel), immediately followed by accelerated concomitant boost radiotherapy (44 Gy in 22 fractions) and definitive surgery. The primary endpoint was event-free survival at 12 months. Efficacy analyses were done by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00030810. FINDINGS: 46 patients were enrolled, with a median age of 60 years (range 28-70). 13 (28%) patients had N3 disease, 36 (78%) had T4 disease. All patients received chemotherapy; 35 (76%) patients received radiotherapy. The main toxicities during chemotherapy were neutropenia (25 patients [54%] at grade 3 or 4) and febrile neutropenia (nine [20%]); the main toxicity after radiotherapy was oesophagitis (ten patients [29%]; nine grade 2, one grade 3). 35 patients (76%) underwent surgery, with pneumonectomy in 17 patients. A complete (R0) resection was achieved in 27 patients. Peri-operative complications occurred in 14 patients, including two deaths (30-day mortality 5.7%). Seven patients required a second surgical intervention. Pathological mediastinal downstaging was seen in 11 of the 28 patients who had lymph-node involvement at enrolment, a complete pathological response was seen in six patients. Event-free survival at 12 months was 54% (95% CI 39-67). After a median follow-up of 58 months, the median overall survival was 29 months (95% CI 16.1-NA), with survival at 1, 3, and 5 years of 67% (95% CI 52-79), 47% (32-61), and 40% (24-55). INTERPRETATION: A treatment strategy of neoadjuvant chemotherapy and radiotherapy followed by surgery is feasible in selected patients. Toxicity is considerable, but manageable. Survival compares favourably with historical results of combined treatment for less advanced stage IIIA disease. FUNDING: Swiss Group for Clinical Cancer Research (SAKK) and an unrestricted educational grant by Sanofi-Aventis (Switzerland).

Identificação e quantificação de fungos micorrízicos arbusculares autóctones em municípios produtores de citros no Rio Grande do Sul

O objetivo deste trabalho foi identificar e quantificar os esporos de FMA presentes em viveiros e pomares de citros no Rio Grande do Sul. Foram coletadas amostras de solo e de raízes de dez viveiros e doze pomares de oito municípios produtores de citros, nas quais avaliaramse conteúdo nutricional, número de esporos por 100 g de solo seco e colonização das radicelas, e determinaramse as espécies autóctones. As espécies, em ordem decrescente de ocorrência, foram: Glomus macrocarpum > Scutellospora heterogama > Acaulospora scrobiculata = Acaulospora birreticulata > Glomus invermaium = Glomus occultum = Entrophospora colombiana > Glomus claroideum = Glomus constrictum > Scutellospora persica.

Cooperation of human tumor-reactive CD4+ and CD8+ T cells after redirection of their specificity by a high-affinity p53A2.1-specific TCR.

Efficient immune attack of malignant disease requires the concerted action of both CD8+ CTL and CD4+ Th cells. We used human leukocyte antigen (HLA)-A*0201 (A2.1) transgenic mice, in which the mouse CD8 molecule cannot efficiently interact with the alpha3 domain of A2.1, to generate a high-affinity, CD8-independent T cell receptor (TCR) specific for a commonly expressed, tumor-associated cytotoxic T lymphocyte (CTL) epitope derived from the human p53 tumor suppressor protein. Retroviral expression of this CD8-independent, p53-specific TCR into human T cells imparted the CD8+ T lymphocytes with broad tumor-specific CTL activity and turned CD4+ T cells into potent tumor-reactive, p53A2.1-specific Th cells. Both T cell subsets were cooperative and interacted synergistically with dendritic cell intermediates and tumor targets. The intentional redirection of both CD4+ Th cells and CD8+ CTL by the same high-affinity, CD8-independent, tumor-specific TCR could provide the basis for novel broad-spectrum cancer immunotherapeutics.

Características vegetativas, fenológicas e produtivas do pessegueiro cultivar Chimarrita enxertado em diferentes porta-enxertos

O objetivo deste trabalho foi avaliar o efeito de diferentes porta-enxertos no desenvolvimento vegetativo, fenológico e produtivo do pessegueiro [Prunus persica (L.) Batsch] cv. Chimarrita. Os porta-enxertos utilizados foram 'Capdeboscq', 'Tsukuba 1', 'Okinawa', 'Aldrighi' e 'GF 305'. As variáveis analisadas foram: diâmetro do tronco, volume da copa, material retirado da poda, época de brotação, época de floração, quantidade de clorofila, área foliar específica, comprimento dos ramos principais, produtividade, massa dos frutos, coloração do fruto, firmeza de polpa, teor de sólidos solúveis totais (SST), acidez titulável (TA), relação SST/AT, fenóis totais e época de maturação. O delineamento experimental foi o de blocos ao acaso, com quatro repetições com cinco plantas cada. Não houve efeito dos porta-enxertos em: início da brotação, início da queda de folha, área foliar específica e maturação e massa de fruto. O diâmetro de tronco, comprimento de ramos, volume de copa, massa verde da poda e quantidade de clorofila foram influenciados mais pelos porta-enxertos 'Capdeboscq', 'Okinawa' e 'Tsukuba 1'. O maior número de flores por ramo foi obtido com o porta-enxerto 'Capdeboscq'. Frutos mais avermelhados foram produzidos com o porta-enxerto 'GF 305', e os vermelho-amarelados com 'Okinawa'.

Prognostic factors affecting long-term outcomes in patients with resected stage IIIA pN2 non-small-cell lung cancer: 5-year follow-up of a phase II study.

The aim was to investigate the efficacy of neoadjuvant docetaxel-cisplatin and identify prognostic factors for outcome in locally advanced stage IIIA (pN2 by mediastinoscopy) non-small-cell lung cancer (NSCLC) patients. In all, 75 patients (from 90 enrolled) underwent tumour resection after three 3-week cycles of docetaxel 85 mg m-2 (day 1) plus cisplatin 40 or 50 mg m-2 (days 1 and 2). Therapy was well tolerated (overall grade 3 toxicity occurred in 48% patients; no grade 4 nonhaematological toxicity was reported), with no observed late toxicities. Median overall survival (OS) and event-free survival (EFS) times were 35 and 15 months, respectively, in the 75 patients who underwent surgery; corresponding figures for all 90 patients enrolled were 28 and 12 months. At 3 years after initiating trial therapy, 27 out of 75 patients (36%) were alive and tumour free. At 5-year follow-up, 60 and 65% of patients had local relapse and distant metastases, respectively. The most common sites of distant metastases were the lung (24%) and brain (17%). Factors associated with OS, EFS and risk of local relapse and distant metastases were complete tumour resection and chemotherapy activity (clinical response, pathologic response, mediastinal downstaging). Neoadjuvant docetaxel-cisplatin was effective and tolerable in stage IIIA pN2 NSCLC, with chemotherapy contributing significantly to outcomes.

Decreased pyramidal neuron size in Brodmann areas 44 and 45 in patients with autism.

Autism is a neurodevelopmental disorder characterized by deficits in social interaction and social communication, as well as by the presence of repetitive and stereotyped behaviors and interests. Brodmann areas 44 and 45 in the inferior frontal cortex, which are involved in language processing, imitation function, and sociality processing networks, have been implicated in this complex disorder. Using a stereologic approach, this study aims to explore the presence of neuropathological differences in areas 44 and 45 in patients with autism compared to age- and hemisphere-matched controls. Based on previous evidence in the fusiform gyrus, we expected to find a decrease in the number and size of pyramidal neurons as well as an increase in volume of layers III, V, and VI in patients with autism. We observed significantly smaller pyramidal neurons in patients with autism compared to controls, although there was no difference in pyramidal neuron numbers or layer volumes. The reduced pyramidal neuron size suggests that a certain degree of dysfunction of areas 44 and 45 plays a role in the pathology of autism. Our results also support previous studies that have shown specific cellular neuropathology in autism with regionally specific reduction in neuron size, and provide further evidence for the possible involvement of the mirror neuron system, as well as impairment of neuronal networks relevant to communication and social behaviors, in this disorder.

Coregulator Control of Androgen Receptor Action by a Novel Nuclear Receptor-Binding Motif

The androgen receptor (AR) is a ligand-activated transcription factor that is essential for prostate cancer development. It is activated by androgens through its ligand-binding domain (LBD), which consists predominantly of 11 α-helices. Upon ligand binding, the last helix is reorganized to an agonist conformation termed activator function-2 (AF-2) for coactivator binding. Several coactivators bind to the AF-2 pocket through conserved LXXLL or FXXLF sequences to enhance the activity of the receptor. Recently, a small compound-binding surface adjacent to AF-2 has been identified as an allosteric modulator of the AF-2 activity and is termed binding function-3 (BF-3). However, the role of BF-3 in vivo is currently unknown, and little is understood about what proteins can bind to it. Here we demonstrate that a duplicated GARRPR motif at the N terminus of the cochaperone Bag-1L functions through the BF-3 pocket. These findings are supported by the fact that a selective BF-3 inhibitor or mutations within the BF-3 pocket abolish the interaction between the GARRPR motif(s) and the BF-3. Conversely, amino acid exchanges in the two GARRPR motifs of Bag-1L can impair the interaction between Bag-1L and AR without altering the ability of Bag-1L to bind to chromatin. Furthermore, the mutant Bag-1L increases androgen-dependent activation of a subset of AR targets in a genome-wide transcriptome analysis, demonstrating a repressive function of the GARRPR/BF-3 interaction. We have therefore identified GARRPR as a novel BF-3 regulatory sequence important for fine-tuning the activity of the AR.

Desempenho de mudas de mirtilo obtidas por micropropagação ou estaquia

O objetivo deste trabalho foi avaliar o desenvolvimento vegetativo e o início do período produtivo de mirtilos (Vaccinium virgatum) propagados por estaquia e micropropagação. No ano de 2009, foi implantado um pomar de mirtilo com mudas de um ano de idade, em espaçamento de 1,3x4,0 m. Utilizaram-se as cultivares Bluegem, Briteblue e Woodard, do grupo "rabbiteye". Foram avaliados os seguintes parâmetros: altura de planta, diâmetro e número de brotações, diâmetro médio dos frutos, frutos colhidos por planta, produção média por planta, produtividade estimada por hectare, massa de matéria fresca por fruto, teor de sólidos solúveis, acidez titulável e pH. Plantas obtidas pela técnica de micropropagação apresentaram maior crescimento vegetativo inicial, em razão do rejuvenescimento causado por este tipo de propagação. O método de propagação não influenciou a qualidade dos frutos. Ao contrário do esperado, as plantas micropropagadas não mostraram atraso no início da produção de frutos e apresentaram produtividade e qualidade de frutos semelhantes às de plantas propagadas por estaquia.

Hábito de frutificação e produção de pereiras sobre diferentes porta‑enxertos

O objetivo deste trabalho foi avaliar o hábito de frutificação e produção de cultivares de pereira (Pyrus communis) enxertadas sobre porta‑enxertos de marmeleiro (Cydonia oblonga) e de pereira (P. calleryana). O experimento foi realizado durante o inverno de 2009 e 2010, em pomar de pereiras conduzido em líder central, em espaçamento 1,0x5,0 m com três cultivares de pereira (Carrick, Packham e Williams) combinadas com um porta‑enxerto de pereira (Clone D6 de P. calleryana) e 15 de marmeleiro. Foram avaliadas as seguintes variáveis: percentagem de dardos, lamburdas, brindilas vegetativas, brindilas floríferas e de bolsas; número total de gemas floríferas; produção por planta; e eficiência produtiva. O hábito de frutificação das cultivares de pereira é influenciado pelos porta‑enxertos, principalmente no que se refere à formação de lamburdas. Em geral, observou-se relação inversa entre percentagem de dardos e de lamburdas, e entre eficiência produtiva e vigor, para todas as combinações entre copa e porta‑enxerto. Para potencializar a produção, o manejo cultural nos pomares das cultivares avaliadas, principalmente por ocasião da poda, deve ser orientado de acordo com o porta‑enxerto utilizado e o hábito de frutificação de cada combinação.