RESUMO Estudou-se o efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e na população de células natural killers (NK) na decídua e na glândula metrial de ratas durante a gestação. Avaliou-se a detecção imunoistoquímica de interferon γ (IFNγ), do fator inibidor de migração (MIF), da interleucina 15 (IL15), do óxido nítrico sintase induzível (iNOS), a marcação com lectina DBA para evidenciação das células NK uterinas DBA+ e a expressão gênica de Ifnγ e Nos2. O hipotireoidismo aumentou o iNOS aos sete dias, a IL15 e o MIF aos 10 e 12 dias, o IFNγ e o MIF aos 14 DG e a expressão dos transcritos gênicos para iNos aos 12 e 19 dias e para Ifnγ aos 14 DG. O hipotireoidismo reduziu a imunomarcação de MIF e lectina DBA aos sete dias, lectina DBA aos 10 e 14 DG, IFNγ aos 12 dias, e a expressão de Ifnγ aos 10 e 19 DG e de iNOS aos 12, 14 e 19 DG, bem como reduziu seus transcritos gênicos aos 10 e 14 DG. Conclui-se que o hipotireoidismo compromete o perfil imunológico na interface materno-fetal ao longo da gestação, particularmente por reduzir o fator anti-inflamatório iNOS e a população de células uNK DBA+.

ABSTRACT The goal of this study was to evaluate the effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats. Interferon gamma (IFNγ), migration inhibitory factor (MIF), interleukin 15 (IL15), inducible nitric oxide synthase (iNOS), and DBA-Lectin labeling for evidence of uNK DBA+ cells in decidua and genetic expression of Ifnγ and iNos by real-time RT-PCR were evaluated. Hypothyroidism increased protein expression of iNOS at 7 days, IL15 and MIF at 10 and 12 days, IFNγ and MIF at 14 DG in the decidua and/or metrial gland and the gene transcripts for iNOS at 12 and 19 days and for Inf at 14 DG. In addition, hypothyroidism reduced the protein expression of MIF and DBA-Lectin at 7 days, DBA-Lectin at 10 and 14 DG, IFNγ at 12 days, and the gene transcript to Ifnγ at 10 and 19 DGs. Hypothyroidism also reduced the protein expression of iNOS at 12, 14 and 19 DG and reduced its gene transcripts at 10 and 14 DGs. It is concluded that hypothyroidism compromises the immunology profile at the maternal-fetal interface throughout pregnancy, particularly by reducing the anti-inflammatory factor iNOS and population of uNK DBA+ cells.

RESUMO: Altas doses de nitrogênio favorecem o crescimento vegetativo do milho e a perda no rendimento de grãos devido ao acamamento de plantas. Este efeito pode ser mitigado pelos reguladores de crescimento. Objetivou-se com este estudo avaliar o efeito do regulador de crescimento trinexapac-ethyl na resposta de híbridos de milho as doses de nitrogênio em cobertura. O experimento foi conduzido em Lages, SC, em dois anos agrícolas, testando dois híbridos (P1630 e P30F53) e quatro doses de nitrogênio em cobertura (0, 150, 300 e 450 kg de N ha-1), com e sem o uso de trinexapac-ethyl. O delineamento experimental foi em blocos casualizados dispostos em parcelas sub-subdivididas, com três repetições por tratamento. Avaliaram-se a altura de plantas e inserção de espiga, o acamamento e quebra de plantas, o rendimento de grãos e os componentes do rendimento. O rendimento de grãos aumentou de forma quadrática com o incremento das doses de nitrogênio. Os máximos rendimentos de grãos foram de 15.784 e 15.257 kg ha-1, em 2014/2015 e 2015/2016, respectivamente. Doses maiores do que 150 kg ha-1 de N, independentemente da utilização do regulador de crescimento, promoveram, no máximo, incrementos no rendimento de grãos de 14,7% a 18,1%. A aplicação de trinexapac-ethyl reduziu o comprimento dos entrenós superiores do colmo e a massa de mil grãos, mas não interferiu na porcentagem de plantas acamadas e no rendimento de grãos. Portanto, o uso de trinexapac-ethyl não aumenta a resposta do rendimento de grãos do milho ao incremento na dose de nitrogênio em cobertura.

ABSTRACT: High nitrogen rates favor maize vegetative growth and grain yield losses due to stem lodging. The use of growth regulators can mitigate such effect. This work was carried out aiming to evaluate the effect of trinexapac ethyl on maize hybrids response to side-dress nitrogen rates. The experiment was carried out in Lages, Santa Catarina State, South of Brazil, during two growing seasons with two hybrids (P1680 and P30F53), and four nitrogen rates (0, 150, 300 and 450 kg N ha-1) were tested, with and without the presence of trinexapac ethyl. The experimental design was in randomized blocks arranged in split-split plots, with three replications per treatment. The plants height and ear insertion height, plant lodging and breaking, grain yield and yield components were evaluated. Grain yield increased in a quadratic way with increased nitrogen rate. The highest grain yields were 15,784 and 15,257 kg ha-1, in 2014/15 and 2015/16, respectively. Application rates higher than 150 kg of N ha-1, regardless of the use of the plant growth regulator, promoted maximum grain yield increases, ranging from 14.7 to 18.1%. The application of trinexapac-ethyl reduced the upper stem internode length, 1000-grain dry weight and did not affect the percentage of lodged plants and grain yield of both hybrids. Therefore, the use of trinexapac-ethyl did not enhance the maize grain yield response to increased rates of side-dress nitrogen.

RESUMO: Na cultura do milho há poucos relatos sobre o uso de trinexapac-ethyl (TE) visando o manejo do acamamento de plantas. Este estudo teve por objetivo avaliar diferentes doses sequenciais de TE aplicadas em diferentes estádios fenológicos da planta de milho, utilizando o híbrido simples P30F53HR e a variedade SCS 154 Fortuna. Os experimentos foram realizados em casa de vegetação em dois anos de cultivo: 2013 e 2014. Foram empregados vasos com capacidade para 5 L, preenchidos com 75% de solo e 25% de substrato comercial, com uma planta de milho por vaso. O delineamento experimental foi de blocos ao acaso com quatro repetições. Os tratamentos consistiram de: (T1) testemunha (sem aplicação de regulador); (T2) aplicação em V2 (100 g i.a. ha-1); (T3) aplicação de 100 g i.a. ha-1 em V2+V3; (T4) aplicação de 100 g i.a. ha-1 em V2+V3+V4; (T5) única aplicação de 300 g i.a. ha-1 em V4; (T6) aplicação de 100 g i.a. ha-1 em V2+V3+V4+V5; (T7) aplicação de 100 g i.a. ha-1 em V2+V3+V4+V5+V6; e (T8) aplicação de 100 g i.a. ha-1 em V2+V3+V4+V5+V6+V7. A partir do estádio V4 as plantas de milho apresentaram sinais de sensibilidade ao regulador de crescimento; a aplicação de V2 a V7 (T8), comparada à de V2 a V6 (T7), evidenciou que as plantas foram mais sensíveis ao TE, com redução superior a 45% na estatura de planta - resposta esta similar entre os dois genótipos de milho. Os resultados indicam que após o estádio V6 as plantas são altamente sensíveis ao trinexapac-ethyl.

ABSTRACT: In Brazilian agriculture there are few reports on trinexapac-ethyl (TE) effects on maize plant lodging management. This study aimed to evaluate the effects of sequential application of TE sprayed on different plant growth stages of maize using the simple hybrid P30F53HR and the variety SCS 154 Fortuna. Experiments were carried out in a greenhouse in the 2013 and 2014 harvests. Plants were grown singly in 5L pots filled with 75% soil and 25% substrate. The experimental design was a randomized complete block with four replications. The following treatments were performed: (T1) control (no application of growth retardant); (T2) application at V2 (100 g a.i. ha-1); (T3) sequential application of 100 g a.i. ha-1 at V2 plus V3; (T4) sequential application of 100 g a.i. ha-1 at V2+V3+V4; (T5) single application of 300 g a.i. ha-1 at V4; (T6) sequential application of 100 g a.i. ha-1 at V2+V3+V4+V5; (T7) sequential application of 100 g a.i. ha-1 at V2+V3+V4+V5+V6 and (T8) sequential application of 100 g a.i. ha-1 at V2+V3+V4+V5+V6+V7. The some morphological characteristics of the two maize genotypes changed in response to TE treatment. From stage V4 onwards, maize plant height showed signs of sensitivity to the growth retardant. Plants in T8 (TE applied over V2 to V7) were more sensitive to TE with over 45% reduction in plant height compared with application T7 (applied from V2 to V6). This response was similar in both maize genotypes, thus indicating that plants are highly sensitive to TE after the V6 stage.

RESUMO Indução e padrão de crescimento de calos de folhas de Piper permucronatum. Piper permucronatum é um arbusto perene, uma planta medicinal native da Floresta Amazônica. Tradicionalmente, o chá de suas folhas é usado em casos de cólicas menstruais e intestinais, dores de estômago, problemas digestivos, diarreia, hemorragia e náusea. O óleo essencial das folhas é efetivo contra a larva de Aedes aegypti; suas flavonas e flavanonas têm efeito fungicida contra Clamidosporium cladosporioides e C. sphaerospermum; seu extrato hexânico é efetivo contra Leishmania amazonensis. O objetivo deste trabalho foi determinar um protocolo para indução de calos em folhas de P. permucronatum e identificar o padrão de crescimento dos calos, com foco na fase de desaceleração, quando as células de calo devem ser subcultivadas em meio líquido para produzir culturas de células em suspensão. Explantes foliares foram inoculados em meio MS sólido suplementado com combinações fatoriais de 2,4-D, BAP, ANA e GA3. A formação de calos foi avaliada semanalmente até o 49º dia. Posteriormente, novos explantes foram inoculados na combinação hormonal que resultou na maior proliferação de células de calo e, a cada sete dias durante 70 dias, amostras foram secas e pesadas para determinar o padrão de crescimento dos calos. ANA e GA3 não foram efetivas para a indução de calos. Combinações de 2,4-D e BAP resultaram em indução e proliferação de calos. A maior porcentagem de indução de calos foi observada com a combinação de 4,52 µM de 2,4-D e 4,44 µM de BAP. Os calos produzidos eram friáveis e esbranquiçados. O crescimento dos calos seguiu um padrão sigmoide. A fase de desaceleração iniciou no 56º dia de cultivo.

ABSTRACT Piper permucronatum is a perennial shrub, a medicinal plant native to the Amazon Rainforest. Traditionally, the tea of its leaves is used to combat menstrual and intestinal cramps, stomach pain, digestive problems, diarrhea, hemorrhage, and nausea. Its leaf’s essential oil is effective against Aedes aegypti larvae; its flavones and flavanones have a fungicidal effect against Clamidosporium cladosporioides and C. sphaerospermum; its hexanic extract is effective against Leishmania amazonensis. The objective of this study was to provide a protocol for callus induction from P. permucronatum leaves and an identification of the callus growth pattern, focusing on the deceleration phase, when the callus cells must be subcultured into liquid medium in order to produce a cell suspension cultures. Leaf explants were inoculated in a solid MS medium supplemented with factorial combinations of 2,4-D, BA, NAA and GA3. Callus formation was evaluated weekly until the 49th day. Subsequently, new explants were inoculated at the hormonal combination that resulted in the highest callus cell proliferation and, every seven days during a period of 70 days, samples were dried and weighed to determine the callus growth pattern. NAA and GA3 were not effective for callus induction. Combinations of 2,4-D and BA resulted in callus induction and proliferation. The highest percentage of callus induction was observed with the combination of 4.52 µM 2,4-D and 4.44 µM BA. The calluses thereby produced were friable and whitish. The callus growth pattern followed a sigmoid shape. The deceleration phase started on the 56th day of culture.

Annona mucosa é uma árvore frutífera da família Annonaceae, produtora de importantes metabólitos secundários de interesse medicinal, como lignanas, acetogeninas e alcaloides. A cultura in vitro de calos representa um importante recurso para a produção contínua de metabólitos, viabilizando a conservação da biodiversidade química e a obtenção controlada de material para estudos biológicos e fitoquímicos. O objetivo deste trabalho foi otimizar a produção de calos friáveis de A. mucosa, avaliando o efeito de diferentes meios nutritivos e fitorreguladores. Segmentos de folha e de hipocótilo de plântulas germinadas in vivo foram utilizados como explantes e inoculados nos meios de cultura MS, WPM e B5 suplementados com picloram (2 - 20µM) isolado ou combinado com as citocininas BAP, KIN ou TDZ (0,2 - 1µM). As culturas foram mantidas a 26±2ºC, no escuro, com subcultivos mensais. A produção de calos foi avaliada por aferição do peso dos calos, após 90 dias. Em todos os tratamentos na presença da auxina picloram, o cultivo de hipocótilos resultou em maior porcentagem de formação de calos, sobretudo no meio de cultura WPM. A associação com TDZ produziu massa calogênica friável altamente proliferativa e ausente de oxidação, alcançando valores superiores àqueles obtidos em trabalhos prévios com a espécie. Os resultados viabilizam o uso do material em suspensões celulares e posterior caracterização fitoquímica para a exploração da produção in vitro de metabólitos da espécie.

The Annona mucosa is a fruit tree of the Annonaceae family that produces a range of secondary metabolites of medicinal interest, such as lignans, acetogenins and alkaloids. The callus culture represents a renewable source of valuable medicinal compounds and controlled supply of material for biological and phytochemical studies. Therefore, this study was carried out to investigate the effects of three nutrient media, different concentrations of picloram and cytokinin types, in order to optimize the biomass yield and friability of calluses of A. mucosa. Leaf and hypocotyl segments from seedlings produced from in vivo seed germination were used as explants, which were inoculated in MS, WPM and B5 culture media supplemented with picloram (2-20µM) only or in addition to the cytokinins BAP, KIN or TDZ (0,2 - 1µM ). Cultures were maintained at 26±2ºC in the dark, with monthly subcultures. After 90 days, biomass production was evaluated. In all treatments, hypocotyl explants provided the highest percentage of callus formation, particularly in WPM. The association with TDZ produced highly proliferative friable callus, with no oxidation, reaching higher values than the previous works with this species. The results enable the use of the calluses produced in cell suspensions and the subsequent phytochemical characterization, in order to explore the in vitro production of metabolites of the species.

A presente pesquisa teve por objetivo estudar a influência de diferentes densidades de plantas e genótipos de soja Roundup ReadyTM (RR) em algumas características morfoagronômicas da soja, semeada em duas safras distintas. O experimento foi conduzido na região dos Campos de Cima da Serra, no Estado do Rio Grande do Sul, nas safras agrícolas 2006/07 e 2007/08, com os cultivares CD 213RR e BRS 255RR, nas densidades de 12, 18, 24, 30 e 36 plantas aptas por m². O delineamento experimental utilizado foi de blocos ao acaso, com quatro repetições. Para isso, foram avaliados altura de plantas, diâmetro da haste, número de ramificações por planta, número de vagens por planta, número de grãos por planta, número de grãos por vagem, massa de mil grãos e rendimento de grãos por hectare. O manejo das plantas daninhas com glyphosate em duas etapas (pré-semeadura e em V4) evitou eficientemente a matocompetição. O aumento da densidade de plantas de soja RR promoveu incremento linear da altura das plantas (0,5 a 0,9 cm por planta m-2), influenciada pelo genótipo e pelo ano de cultivo. O diâmetro da haste foi diminuído com o aumento da densidade de plantas de soja RR de forma linear (-0,03 a -0,2 mm por planta m-2). O número de ramos por planta reduziu com o aumento da densidade (-0,05 a -0,19 ramos por planta). Na segunda safra, o rendimento de grãos e a massa de mil grãos foram menores, afetados por um período de estiagem e também por uma geada no final do ciclo, mais acentuadamente para o CD 213RR. A combinação de maior número de plantas m-2 levou a uma redução do número de vagens por planta e de grãos por vagem, embora de magnitudes distintas entre as cultivares, o que foi discutido pela interação entre os fatores; no entanto, o rendimento de grãos não foi alterado nas diferentes densidades testadas, e sim sofreu influência da safra de cultivo.

This work studied the influence of different plant densities and Roundup ReadyTM (RR) soybean genotypes, on some morphoagronomic characteristics of soybean sown in two distinct crops. The experiment was carried out in Campos de Cima da Serra in the state of Rio Grande do Sul, Brazil, in the 2006/07 and 2007/08 growing seasons with CD 213RR and BRS 255RR cultivars, at densities of 12, 18, 24, 30 and 36 suitableplantsm-2. The experimental layout used was randomized blocks with four repetitions. Plant height, stem diameter, number of ramifications per plant, number of pods per plant, number of grains per plant, number of grains per pod, mass of a thousand grains and yield of grains per hectare were evaluated. Weed plant management with glyphosate in two applications (pre-sowing and V4) efficiently prevented weed competition. Increasing RR soybean plant density led to a linear increase in plant height (0.5 to 0.9 cm per plant m-2), influenced by genotype and crop year. Stem diameter reduced linearly with an increase in plant density (-0.03 to -0.2 mmper plantm-2). The number of ramifications per plant decreased with an increase in density (-0.05 to -0.19 branch per plant). In the second harvest, grain yield and mass of a thousand grains were less affected by drought and frost at the end of the crop cycle, more accentuated in cultivar CD 213RR. A combination of a greater number of plantsm-2 reduced the number of pods per plant and grains per pod, though by different magnitudes between cultivars, explained by the interaction among various factors. However, grain yield was not affected by the different densities tested, being rather influenced by the growing season.

Interleukin-18 (IL-18) is a cytokine that belongs to the IL-1 family. Endometriosis is strongly associated with sub-fertility, and affects about 15% of women of reproductive age. IL-18 may favor the progression of endometriosis. The objective of the present study was to determine the concentration of IL-18 in the serum and peritoneal fluid of infertile women with endometriosis. Forty infertile and 25 fertile women were screened in a teaching hospital. Thirty-four infertile patients with minimal or mild endometriosis and 22 fertile controls were enrolled in the study. The primary outcome was the estimate of IL-18 levels and the secondary outcome was the correlation between serum and peritoneal levels of IL-18. There were no differences between the two groups regarding age, body mass index and levels of peritoneal fluid IL-18 (mean ± SD): 290.85 ± 173.02 pg/mL for infertile women vs 374.21 ± 330.15 pg/mL for controls; or serum IL-18: 391.07 ± 119.71 pg/mL for infertile women vs 373.42 ± 129.11 pg/mL for controls. However, a positive association was found between serum and peritoneal IL-18 levels in patients with endometriosis: r = 0.794, P = 0.0001. All measurements were carried out at the same time by the Human IL-18 Immuno Assay ELISA kit (MBL Co. Ltd., Japan). The present study did not find evidence supporting the hypothesis that IL-18 levels are associated with infertility in women with minimal and mild endometriosis, although a positive correlation was detected in these women between peritoneal and serum levels of IL-18.

Apoptotic protease activating factor 1 (APAF-1) has a critical role in the regulation of apoptosis. In the present study, the mRNA expression analysis of different APAF-1 transcripts (APAF-1S, APAF-1LC, APAF-1LN, and APAF-1XL) was analyzed in bone marrow samples from 37 patients with acute myeloid leukemia (newly diagnosed, with no previous treatment). APAF-1XL and APAF-1LN transcripts (with and without an extra WD-40 repeat region, respectively) were detected in all samples, although the major form expressed was APAF-1XL in 65% of the samples (group 1), while 35% of the samples expressed primarily APAF-1LN (group 2). Only 46% of the patients presented complete remission in response to remission induction therapy (represented by less than 5% marrow blasts and hematological recovery), all but 2 cases being from group 1, 21.6% did not attain complete remission (only 1 case from group 1), and 32.4% of the patients died early. Lower expression of APAF-1XL (APAF-1XL/APAF-1LN ratio <1.2) was associated with a poor response to therapy (P = 0.0005, Fisher exact test). Both groups showed similar characteristics regarding white blood cell counts, cytogenetic data or presence of gene rearrangements associated with good prognosis as AML1-ETO, CBFB-MYH11 and PML/RARA. Since it has been shown that only the isoforms with the extra WD-40 repeat region activate procaspase-9, we suggest that low procaspase-9 activation may also be involved in the deregulation of apoptosis and chemotherapy resistance in acute myeloid leukemia.

Urethrostomy for serial sampling of urine was used due to the necessity of one technique that allowed the total urinary collection in definite time periods. Twenty-seven male Holstein calves from 8 to 16-days-age having a mean weight 39.50±4.80kg were used. Urethrostomy was carried out with the use of urethral sounding lead that was made for this purpose. In agreement with the clinical examinations, including seric biochemical, urinalysis and blood test and postmortem examination, the technique achieved 81.5% of satisfactory results.

The feasibility of allogeneic bone marrow transplantation (alloBMT) in a developing country has not yet been demonstrated. Many adverse factors including social and economic limitations may reduce the overall results of this complex and expensive procedure. Our objective was to characterize the most important clinical, social and economic features of candidates for transplantation and their potential donors as well as the influence of these factors on overall survival in a retrospective and exploratory analysis at a university hospital. From July 1993 to July 2001, candidates for BMT were referred to the Bone Marrow Transplantation Unit by Hematology and Oncology Centers from several regions of Brazil. A total of 1138 patients were referred to us as candidates for alloBMT. Median age was 25 years (range: 2 months-60 years), 684 (60.1%) were males and 454 (39.9%) were females. The clinical indications were severe aplastic anemia and hematological malignancies. From the total of 1138 patients, 923 had HLA-typing; 497/923 (53.8%) candidates had full match donors; 352/1138 (30.8%) were eligible for alloBMT. Only 235 of 352 (66.7%) were transplanted. Schooling was 1st to 8th grade for 123/235 (52.3%); monthly family income ranged from US$60 (7%) to more than US$400 (36%). Overall survival for patients with chronic myeloid leukemia, severe aplastic anemia and acute myeloid leukemia was 58, 60 and 30%, respectively. Thus, overall survival rates for the most frequent hematological diseases were similar to those reported in the International Registry, except for acute myeloid leukemia. This descriptive and exploratory analysis suggests the feasibility of alloBMT in a developing country like Brazil.

The purpose of the present study was to evaluate the mixed lymphocyte culture as a predictive assay of acute and chronic graft-versus-host disease (GVHD). We studied 153 patients who received a first bone marrow transplantation from human leukocyte antigen-identical siblings. Acute GVHD was observed in 26 of 128 (20.3%) patients evaluated and chronic GVHD occurred in 60 of 114 (52.6%). One-way mixed lymphocyte culture (MLC) assays were performed by the standard method. MLC results are reported as the relative response (RR) from donor against patient cells. The responses ranged from -47.0 to 40.7%, with a median of 0.5%. The Kaplan-Meier probability of developing GVHD was determined for patients with positive and negative MLC. There was no significant difference in incidence of acute GVHD between the groups studied. However, the incidence of chronic GVHD was higher in recipients with RR >4.5% than in those with RR <=4.5%. The Cox Proportional Hazards model was used to examine the effect of MLC levels on incidence of chronic GVHD, while adjusting for the potential confounding effect of others suspected or observed risk factors. The relative risk of chronic GVHD was 2.5 for patients with positive MLC (RR >4.5%), 2.9 for those who received peripheral blood progenitor cells as a graft, and 2.2 for patients who developed previous acute GVHD. MLC was not useful for predicting acute GVHD, but MLC with RR >4.5% associated with other risk factors could predict the development of chronic GVHD, being of help for the prevention and/or treatment of this late complication.

Trials have demonstrated that high-dose escalation followed by autologous transplantation can promote better long-term survival as salvage treatment in malignant lymphomas. The aim of the present nonrandomized clinical trial was to demonstrate the role of high-dose cyclophosphamide (HDCY) in reducing tumor burden and also to determine the effectiveness of HDCY followed by etoposide (VP-16) and methotrexate (MTX) in Hodgkin's disease plus high-dose therapy with peripheral blood progenitor cell (PBPC) transplantation as salvage treatment. From 1998 to 2000, 33 patients with a median age of 33 years (13-65) affected by aggressive non-Hodgkin's lymphoma (NHL) (60.6%) or persistent or relapsed Hodgkin's disease (39.4%) were enrolled and treated using high dose escalation (HDCY + HDVP-16 plus HDMTX in Hodgkin's disease) followed by autologous PBPC transplantation. On an "intention to treat" basis, 33 patients with malignant lymphomas were evaluated. The overall median follow-up was 400 days (40-1233). Thirty-one patients underwent autografting and received a median of 6.19 x 10(6)/kg (1.07-29.3) CD34+ cells. Patients who were chemosensitive to HDCY (N = 22) and patients who were chemoresistant (N = 11) presented an overall survival of 96 and 15%, respectively (P<0.0001). Overall survival was 92% for chemosensitive patients and 0% for patients who were still chemoresistant before transplantation (P<0.0001). Toxicity-related mortality was 12% (four patients), related to HDCY in two cases and to transplant in the other two. HDCY + HDVP-16 plus HDMTX in only Hodgkin's disease followed by autologous PBPC proved to be effective and safe as salvage treatment for chemosensitive patients affected by aggressive NHL and Hodgkin's disease, with acceptable mortality rates related to sequential treatment.

Clinical trials indicate that amifostine may confer protection on various normal tissues without attenuating anti-tumor response. When administered prior to chemotherapy or radiotherapy, it may provide a broad spectrum of cytoprotection including against alkylating drugs. The mechanism of protection resides in the metabolism at normal tissue site by membrane-bound alkaline phosphatase. Toxicity of this drug is moderate with hypotension, nausea and vomiting, and hypocalcemia being observed. We report a phase II study using amifostine as a protective drug against high-dose cyclophosphamide (HDCY) (7 g/m2), used to mobilize peripheral blood progenitor cells (PBPC) and to reduce tumor burden. We enrolled 29 patients, 22 (75.9%) affected by aggressive and 7 (24.1%) by indolent non-Hodgkin's lymphoma (NHL), who were submitted to 58 infusions of amifostine and compared them with a historical group (33 patients) affected by aggressive NHL and treated with VACOP-B followed by HDCY. The most important results in favor of amifostine were the reduction of intensity of cardiac, pulmonary and hepatic toxicity, and a significant reduction of frequency and severity of mucositis (P = 0.04). None of the 29 patients died in the protected group, while in the historical group 2/33 patients died because of cardiac or pulmonary toxicity and 2 patients stopped therapy due to toxicity. Amifostine did not prevent the aplastic phase following HDCY. PBPC collection and hematological recovery were adequate in both groups. The number of CFU-GM (colony-forming units-granulocyte/macrophage) colonies and mononuclear cells in the apheresis products was significantly higher in the amifostine group (P = 0.02 and 0.01, respectively). Side effects were mild and easily controlled. We conclude that amifostine protection should be useful in HDCY to protect normal tissues, with acceptable side effects.