A limited antibody panel can distinguish B-precursor acute lymphoblastic leukemia from normal B precursors with four color flow cytometry: implications for residual disease detection.

Multiparameter flow cytometry may be used to detect minimal residual disease in acute leukemia because leukemic cells often display aberrant phenotypes when compared to normal cells. One limitation of this approach in B-precursor ALL is that leukemic phenotypes are often qualitatively similar to normal marrow B progenitors, though it has long been recognized that the latter show a predictable pattern of antigen expression with differentiation. In this study we used four-color flow cytometry to define precisely the patterns of normal antigen expression on a series of normal bone marrows using two different four-color combinations of antibodies: CD19-APC/CD45-perCP/CD20-PE/CD10-FITC; and CD19-APC/CD45-perCP/CD9-PE/CD34-FITC. A series of dual parameter displays were created in which normal B precursors occupied predictable regions. We then tested these antibody combinations on a series of 82 cases of B-precursor ALL and found that in 76/82 cases (93%) the first combination demonstrated an abnormal population on at least one of the dual parameter displays, and that 72/77 cases tested (94%) showed an abnormality with the second combination. When taken together, 81/82 cases (99%) showed an abnormality. When purified blasts were serially diluted into normal marrows we found a sensitivity of detection of 1 cell in 10(4) normal marrow cells provided sufficient CD19+ cells were acquired to visualize the abnormal population as a discrete cluster. Because the pattern of antigen expression in normals is very reproducible, it is possible to create a fixed set of geometrical regions to define the normal; this makes analysis of an unknown sample very straightforward. We conclude that our approach could be employed as a simple method for the detection of minimal residual disease in B-precursor ALL, and unlike many other methods should prove applicable to virtually all cases of this malignancy.

Multiparameter flow-cytometric analysis of bcl-2 and Fas expression in normal and neoplastic hematopoiesis.

Apoptosis (programmed cell death) is an important regulatory mechanism in hematopoiesis, and is thought to be a principal mechanism of action of cytotoxic chemotherapy. Proteins that modulate apoptosis include bcl-2, which inhibits apoptosis, and Fas (CD95, also known as APO-1), which induces apoptosis in susceptible cells bound by Fas ligand (FasL). To characterize precisely the expression of these apoptosis-regulatory proteins in normal and neoplastic hematopoiesis, the authors have performed multiparameter flow cytometric (FCM) analysis in a series of normal and abnormal marrow specimens. Among normal hematopoietic elements, bcl-2 expression was highest in myeloblasts (29 [+/- 9] x 10(3) molecules of equivalent soluble fluorochrome [MESF]), and lymphocytes (28[+/- 7] x 10(3) MESF). bcl-2 was essentially undetectable in granulocytes and nucleated red blood cells, whereas monocytes and B-cell precursors expressed intermediate levels of bcl-2 (11[+/- 4] x 10(3) and 7[+/- 1] x 10(3) MESF, respectively). Fas expression increased with granulocytic and monocytic differentiation; myeloblasts expressed 8(+/- 2) x 10(3) MESF, whereas granulocytes (15 [+/- 2] x 10(3) MESF) and monocytes (28[+/- 5] x 10(3) MESF) displayed relatively greater intensity of staining for Fas. Among lymphoid cells, Fas expression was heterogeneous. B cells expressed lower intensity Fas staining than both CD4+ and CD8+ T cells. Myeloblasts in 30 cases of myeloid leukemia and myelodysplasia studied for bcl-2 and/or Fas expression manifested variable levels of these molecules (range 9-105 x 10(3) MESF for bcl-2 and 3-33 x 10(3) MESF for Fas). In addition, intraclonal heterogeneity of bcl-2 and Fas expression was seen in certain cases of AML, which correlated with extent of differentiation. Among 28 cases of B-precursor ALL studied for bcl-2 and/or Fas expression, bcl-2 ranged from 22 to 60 x 10(3) MESF (P < .001 versus normal marrow B-cell precursors), and Fas varied between essentially undetectable levels and 6 x 10(3) MESF. In summary, normal marrow subsets display characteristic levels of the apoptosis-regulatory molecules, bcl-2 and Fas. In hematopoietic neoplasms, expression of bcl-2 and Fas varies among cases, and in some instances, within leukemic blast populations. Further study is required to understand the potential significance of this heterogeneous expression of bcl-2 and Fas in hematologic neoplasia.

Comparison of conventional and ThinPrep preparations of mucoid cytology samples.

Cytological specimens containing large amounts of mucus and/or blood are often a considerable problem as far as preparation and especially screening are concerned. The ThinPrep method, which uses a mucolytic and hemolysing solution and prepares slides automatically, eliminates difficulties in both areas. Comparing ThinPreps to conventionally prepared samples reveals equivalent or even superior cellular preservation. Cells are spread in a thin layer, mostly with only a minimal amount of residual mucus or blood. Screening is consequently much easier and faster. The time-saving effect does not compromise the diagnostic accuracy, as we could demonstrate with an 92% overall agreement in both procedures. The difference of 8% could possibly be attributed to inadequate splitting of the samples.

[Tumor of the maxillary sinus with orbital invasion]. / Tumeur du sinus maxillaire avec envahissement orbitaire.

The authors report the case of an epidermoid carcinoma, originating from the maxillary sinus, with orbital extension. The first harmless symptoms can mislead the diagnosis. Other signs will appear a few weeks later leading to a tumoral process. The further examinations will confirm the fears.

The versatility of the sub-mental flap: a case report.

The sub-mental flap has been used in four elderly patients (mean age: 83 years) for reconstruction of defects after oncological resection: three had basal cell carcinoma (cheek, temporal region and fronto-temporal region). One had a squamous cell carcinoma of the hard palate. We believe that the latter example is original and present it in this article. This case shows that the sub-mental flap in addition to its intrinsic qualities is a reliable flap which may be useful in difficult repairs. It can be used to repair wide palatal fistula which occurs after oncological resections.

[Traumatic facial arteriovenous fistula]. / Fistules artério-veineuses faciales traumatiques.

This paper gives a brief description of the aetiology, pathophysiology, diagnosis and treatment of traumatic arteriovenous communications. Traumatic arteriovenous communications of the face and scalp are rare lesions characterized by multiple endothelial-lined channels between the arterial and venous system. A swelling of the face with pulsatile tinnitus comprise the main presenting symptoms. Lesions should be delimited by arteriography unless small and localized. They are managed by complete excision and ligation of arterial feeding vessels. One case of traumatic arteriovenous communication of the face is reported.