Jean-François Bernaudin

Pulmonary complications occurring after allogeneic bone marrow transplantation. A study of 130 consecutive transplanted patients

This report deals with 81 pulmonary episodes occurring in 130 consecutive patients who underwent allogeneic bone marrow transplantation for hematologic malignancy in the same unit over a 5‐year period. These episodes observed in 69/130 patients (53%) were mostly of infectious origin, and were investigated by bronchoalveolar lavage (BAL). The main causes of pneumonia were: cytomegalovirus (CMV) (n = 25), bacterial pneumonia (n = 17), invasive aspergillosis (n = 11) and pulmonary hemorrhage (n = 9). The overall mortality due to or associated with pneumonia was 26/130 (20%). Graft‐versus‐host disease clearly increased the incidence of infectious pneumonia and the mortality due to or associated with pneumonia. Granulocyte transfusions did not influence the incidence of CMV pneumonitis. The main causes and risk factors for pneumonia are discussed. The role of BAL as a noninvasive procedure is stressed. Cancer 58:1047‐1054, 1986.

Bronchoalveolar lavage during neutropenic episodes: diagnostic yield and cellular pattern

Few data are available concerning the relationship between alveolar and blood cell populations during neutropenia. We wanted to compare the value of pulmonary endoscopic procedures with lavage in neutropenic (polymorphonuclear (PMN) count < or = 1,000.mm-3) and non-neutropenic settings. We therefore, retrospectively, reviewed the results of 118 investigations for pneumonia in patients with malignant haematological diseases. All had bronchoalveolar lavage (BAL), and some had additional studies with protected bacteriological samples. Each BAL specimen was studied after cytocentrifugation by cytological examination for opportunistic infections, haemorrhage, virus, legionellae, and bacteriological cultures. The diagnostic yield of all endoscopic procedures (BAL, telescoping plugged catheter and protected specimen brush) was 53% in neutropenic (Group 1) and 61% in non-neutropenic (Group 2) patients. The aetiological pattern of pneumonia was nearly the same in the two groups, except for more alveolar proteinosis in Group 1 and more cytomegalovirus (CMV) in Group 2. The absolute number of alveolar cells recovered through BAL (total number, macrophages, lymphocytes and PMNs) was significantly lower in neutropenic patients. We conclude that: 1) neutropenic patients with pneumonia require the same investigative approach as non-neutropenic patients; 2) profound neutropenia may be concomitant with a decreased cellularity of alveoli, which may reflect the consequences of marrow aplasia on the pulmonary cell population and/or direct effect of chemotherapy on the lung.

Rat pleural mesothelial cells in culture

SummaryA culture system has been developed for long-term maintenance of rat pleural mesothelial cells. Mesothelial cells were isolated from the parietal pleura of rats and cultured in NCTC 109 medium supplemented with 10% fetal bovine serum. The cell explants attached to the dish and formed a confluent monolayer of polygonal cells within 10 to 15 days. Subcultures were made in the same medium. The mean population doubling time was approximately 30 hr. The ultrastructure of the mesothelial cells in culture was studied by light and electron microscopy and was compared with that of cells obtained from submesothelial components.

Expression of the c-fos proto-oncogene in bone, cartilage and tooth forming tissues during mouse development

The c‐fos proto‐oncogene is the cellular homologue of v‐fos identified as the bone transforming gene of the FBJ and the FBR murine osteosarcoma viruses. We show here, using a sensitive in situ hybridization method, that the c‐fos proto‐oncogene is expressed in the cartilage, bone and tooth forming tissues during mouse development. This result suggests that the tumors observed after infection by the FBJ viral complex and c‐fos overexpression in transgenic mice occur in those tissues in which c‐fos is expressed during development.

Effects of Infused Intralipids on Neutrophil Chemotaxis during Total Parenteral Nutrition

A number of previous studies have suggested that the fat emulsion, Intralipid, might compromise human host defenses, due mainly to impairment of neutrophil functions. The aim of this study was to evaluate the effects of Intralipid on neutrophil chemotaxis in cancer patients receiving total parenteral nutrition including 500 ml of 20% Intralipid over 6 hours (83 ml/hr). No impairment of neutrophil chemotaxis was found during or after lipid infusion. Further investigations are necessary to determine whether, in routine clinical practice, intralipids are responsible for impairment of other neutrophil functions and whether side treatments have a protective effect for neutrophil functions.

Can the centrosome be a marker for DNA ploidy in breast cancer

Background: The role of DNA ploidy in genomic instability of cancer cells and prognosis has been described in a number of studies. The role of the centrosome in cell cycle has also been reported. Aim: In this study, we aimed to investigate the correlation between the centrosome and DNA ploidy in breast cancer in a search for a cytologic predictive and prognostic marker. Materials and Methods: Cell prints were prepared from cell culture of mesothelial cells, fibroblast cell line MRC5 and breast cancer cell lines MCF7 and T47D. Indirect immunofluorescence was used with anti-γ-tubulin and centrosomes were quantified using a fluorescence microscope. DNA ploidy was scored with the DNA index analyzed by flow cytometry. Results: The normal mesothelial cells (94% of the cells with one detected centrosome) and MRC5 diploid cells (68% with two centrosomes) were used as quality controls. A correlation between the number of centrosomes and DNA ploidy was found in MCF7 cell lines (64% of the cells with a number of centrosomes ≥ 3). It was not observed in invasive breast cancer samples; however, the frequency of cells with centrosomes ≥ 3 was found to be slightly higher in DNA aneuploid samples than in DNA diploid samples (15% vs 13.3%). Conclusion: Quantification of centrosome appears to be correlated to DNA ploidy in breast cancer cell lines and slightly associated to DNA aneuploidy in invasive breast cancer. Studies analyzing a larger number of samples as well as morphological abnormalities of the centrosome are needed.

The Pleura: Mesothelial Cells and Mesothelioma

Whereas cell biology and biochemistry of the lung parenchyma is now well documented, little work has been carried out on the pleura. The biological reactivity of pleural cells and the physiology of the pleural space however are of paramount importance, in relation to three main conditions of the pleura: effusions, fibrosis and cancer. The highly significant association of these conditions with past exposure to airborn mineral fibres explains the interest of many workers in pleura, particularly in the field of experimental and human pathology and, more recently that of basic research. According to the work carried out in our laboratory and in others, the structure and function of the pleura will be examined in four parts: 1 - pleural cells in situ and in vitro; 2 - mesothelial barrier, in relation to fluid movement and particulate matters clearance; 3 - inflammatory lesions of the pleura; 4 - pleural carcinogenesis.

Corrélation entre les anomalies centrosomiques et l’ADN-ploïdie dans le cancer du sein

Plusieurs etudes decrivent le role de l’ADN-ploidie dans les instabilites genomiques des cellules cancereuses et le pronostic. Le centrosome jouant un role important dans le cycle cellulaire, de nouveaux facteurs pronostiques pourront etre introduits pour mieux cibler les traitements. L’objectif de ce travail est de rechercher dans le cancer du sein une anomalie centrosomique et la correlation avec l’ADN-ploidie. Des lames sont realisees a partir des echantillons de prelevements mammaires et des suspensions cellulaires (cellules mesotheliales et lignees cellulaires MRC5, MCF7, T47D). L’immunofluorescence indirecte est utilisee avec l’anticorps anti-γ-tubuline puis un fragment F(ab′) 2 conjugue au fluoro-isothyanate. Les centrosomes sont quantifies au microscope a fluorescence. L’ADN-ploidie est definie par l’index d’ADN analysee par cytometrie en flux. Les cellules mesotheliales normales (94 % des cellules avec un centrosome) et les cellules MRC5 diploides cyclantes (68 % avec deux centrosomes) sont des controles de qualite des experiences. Une correlation entre l’ADN-ploidie et le nombre de centrosomes est retrouve dans les lignees MCF7 (64 % des cellules avec un nombre de centrosomes ≥ 3) mais pas dans les lignees T47D et les dix echantillons de carcinome invasif du sein analyses dans cette etude. L’absence de relation entre l’ADN-ploidie et le nombre de centrosomes pourrait etre due au petit nombre d’echantillons mammaires etudies, aux empreintes cellulaires ou au mecanisme tumoral. Un elargissement de l’etude sur un grand nombre d’echantillons et de pathologies mammaires et l’etude des anomalies morphologiques du centrosome sont prevus.ADN ploidy was shown to play a role in genomic instability of cancer cells and prognosis. The implication of the centrosome in the cell cycle was also described. Therefore, new prognostic factors could be suggested for a better-tailored therapy. The purpose of this study is to search for correlation between centrosomal abnormality and ADN ploidy in breast cancer. Cell prints were prepared from cell culture of mesothelial ascitis, fibroblast cell line MRC5 and breast cancer cell lines MCF7 and T47D. Fresh cell prints were also obtained from cases with invasive carcinoma. The centrosome was labelled by an indirect immunofluorescence assay using anti-γ-tubulin antibody and F(ab)(2) FITC before quantification with fluorescence microscopy. ADN ploidy was scored with DNA index obtained by means of flux cytometry. The normal mesothelial cells (94% of cells with only one centrosome) and the diploid cell line MRC5 (68% of cells with two centrosomes) were used as controls. DNA ploidy was found to be correlated with centrosomal abnormality in MCF7 cell line (64% of cells had more than three centrosomes) but not in the 10 cases of invasive ductal carcinoma analysed in this study. The absence of correlation between DNA ploidy and centrosomal abnormality in breast cancer samples may be due to the small numbers of cases, the cell prints or tumorigenesis. Correlation analysis of a larger number of cases and types of breast lesions to numerical and morphological abnormalities of the centrosome are ongoing.