CD5 negative diffuse mantle cell lymphoma with splenomegaly and bone marrow involvement.

We report the case of a 78-year-old man in whom routine physical examination revealed cervical adenopathy and splenomegaly. Peripheral blood showed a normal white blood cell count with an absolute lymphocytosis, which included a population with slightly indented nuclei. Lymph node biopsy showed morphology compatible with mantle cell lymphoma. Bone marrow biopsy showed replacement by a lymphoid proliferation composed of lymphocytes with features similar to those found in the peripheral blood. Immunophenotypic analysis of both peripheral blood and lymph node showed positivity for CD19, CD20 and CD22, with lambda light chain restriction. Tests for CD5 and CD10 were negative. Cytogenetic analysis and polymerase chain reaction studies confirmed the presence of t(11,14) supporting a diagnosis of mantle cell lymphoma. This unusual case of CD5-negative mantle cell lymphoma exemplifies the importance of combined molecular, cytogenetic, and morphologic evaluation when confronted with a lymphoma having an atypical phenotype.

Isodicentric Y chromosome: cytogenetic, molecular and clinical studies and review of the literature.

Dicentrics are among the most common structural abnormalities of the human Y chromosome. Predicting the phenotypic consequences of different duplications and deletions of dicentric Y chromosomes is usually complicated by varying degrees of mosaicism (45,X cell lines), which may, in some cases, remain undetected. Molecular studies in patients with dicentric Y chromosomes have been few, and only two studies have attempted to determine the presence of SRY (the putative testis-determining factor gene). We report an 18-year-old female with short stature, amenorrhea, hirsutism, hypoplastic labia minora, and clitoromegaly who has a 45,X/46,X,idic(Y)(p11.32)/47,X,idic(Y)(p11.32),idic(Y) (p11.32) karyotype. Southern analysis using Y-specific probes (Y97, 2D6, 1F5, pY3.4) and polymerase chain reaction (PCR) analysis using primers for ZFY and SRY were positive for all loci tested, indicating that almost all of the Y chromosome was present. Our findings and an extensive review of the literature emphasize the importance of molecular analyses of abnormal Y chromosomes before any general conclusions can be reached concerning the relative effects of the Y-chromosome abnormality and mosaicism on sexual differentiation.

De novo dup (5p) in a patient with congenital hypoplasia of the adrenal gland.

We report on a black male child with congenital hypoplasia of the adrenal gland (CHA) with a de novo duplication of 5p [dir dup(5) (p13.3-->p15.1)], confirmed by fluorescence in situ hybridization (FISH). In addition to a characteristic clinical course, the patient has hyperpigmentation (melanoderma) since birth, normal external genitalia, marked elevation of ACTH, and absent response to an IV ACTH challenge. To the best of our knowledge, this is the first case of congenital hypoplasia of the adrenal gland associated with a chromosome abnormality. Reviews of dup (5p) and of our patient suggest that duplication of 5p13.3-pter has only minor phenotypic effect, while duplication of the relatively small critical segment p11-p13.2 apparently causes far more deleterious changes. The concurrence of CHA and dup(5p) in our patient may indicate the possible gene localization of an autosomal form of CHA to either at or near 5p13.3 or 5p15.1.

Confirmation of proximal 1q duplication using fluorescence in situ hybridization.

We report on a boy with excessively wrinkled skin, mild micro/brachycephaly with mild hydrocephalus and slightly small temporal lobes, apparently low-set ears, retro/micrognathia and cleft soft palate (Pierre-Robin anomaly), patent ductus arteriosus and foramen ovale, pulmonary hypoplasia, eventration of the left hemidiaphragm, right cryptorchidism, a sacral dimple, flexion contractures of fingers and knees, and equinovarus deformities of both feet. The infant had a de novo dir dup(1)(pter-->q25::q12-->qter). Partial duplications involving proximal 1q have rarely been reported. Furthermore, this is the first case of proximal duplication of chromosome 1q with unequivocal identification using fluorescence in situ hybridization (FISH) with a chromosome 1 painting probe.

Stimulation of growth of a colon cancer cell line by gastrin.

The trophic effects of the hormone gastrin-17 were examined on a human colon cancer cell line. LoVo cells were obtained from the American Type Culture Collection and grown in minimal essential medium in the presence of 10% bovine fetal serum. To demonstrate the trophic effect of gastrin, synchronization was necessary. The effect of gastrin was optimal after 26-h exposure to 0.6 mM thymidine. In the presence of serum the optimal dose of gastrin for stimulation of DNA synthesis was 7.2 X 10(-10) M. Under these conditions gastrin caused a 220% increase in [3H]thymidine incorporation. In the absence of serum the optimal dose of gastrin (3.6 X 10(-9) M) increased DNA synthesis approximately 200%. Twenty-four hours after gastrin treatment (1.8 X 10(-10) M gastrin 17) cell numbers increased 50.8% compared with control. At 48 h this increase was maintained at 44%. Maximum stimulation by gastrin occurred 7-8 h after release from synchronization and exposure to gastrin. This corresponded to the S phase of the cell cycle. Significant stimulation occurred a second time at 22-24 h, presumably during the second S phase in a still synchronous or partially synchronous cell population. These data demonstrate that physiological concentrations of gastrin-17 can stimulate the growth of a human cancer cell line and that some degree of synchronization may be necessary to demonstrate similar effects in other cell lines. Such cell lines may provide a source of rapidly growing cells in which the mechanisms of the trophic effect of gastrin can be examined.

Cytogenetic study of a Merkel cell carcinoma.

Banded karyotypes from two metastatic sites of a Merkel cell carcinoma were analyzed. The results indicate very close similarity between the two specimens including an overall tendency to triploidy and the same five marker chromosomes. One of the markers represented by two copies in most of the karyotyped cells was 1q-.

Progressive chromosome changes associated with different sites of one ovarian carcinoma.

Karyotype analyses were done on cells from the primary and metastatic sites, as well as on the ascitic fluid, from 1 patient with serous carcinoma of the ovary. An increase in the proportion of near-tetraploid cells occurred in the following order: cells from the primary site less than cells from the metastatic site less than cells from ascitic fluid. In addition, two new marker chromosomes appeared among the polyploid cells of ascitic fluid.

Improved method for cytogenetic studies of solid tumors.

Solid human tumors were dissociated with collagenase, cultured for 16-48 hours, and harvested for cytogenetic preparation. Of the 19 tumors used, 14 showed sufficient numbers of metaphases to be useful for chromosome analysis.

Induction of high frequencies of endoreduplication in mammalian cell cultures with 33258 Hoechst and rubidazone.

Cells of the mouse L strain and the Chinese hamster CHO line were treated with 33258 Hoechst, rubidazone, and a combination of these. Recovering cell populations following the drug removal exhibited a high frequency (20--50%) of metaphases with diplochromosomes (endoreduplication), especially in the combination treatment series (up to more than 70% in the L strain). Such a procedure should be useful in probing the mechanisms of the endoreduplication process.

Characterization of an ovarian carcinoma cell line.

A long-term ovarian carcinoma cell line is described in terms of its morphology, original histopathology, electron microscopic features and chromosome features before and after transplantation into an athymic nude mouse. The microscopic features of the tumor nodule grown in the nude mouse are compared with the original pathology. Epithelial characteristics of the cells were maintained in culture and after retrieval from the nude mouse. Three markers were identified in all karyotypes and trisomy was noted in chromosomes 1, 2, 3, 6, 11, 12, 16, and X and monosomy of 17 and 21.

Chromosome banding patterns of L1210 leukemia of the mouse.

The chromosomal characteristics of 2 L1210 mouse leukemia lines, 1 grown in vivo and 1 in vitro, were compared. The procedures used were: G-banding, C-banding and silver staining for nucleolar organizer regions. The 2 lines are not exactly the same, but share some cytogenetic characteristics.

Adriamycin-induced chromosome damage: elevated frequencies of isochromatid aberrations in G2 and S phases.

Analysis of chromosomes from cells treated with adriamycin during G2 and S phases showed a high frequency of isochromatid-type of breaks, in addition to the expected chromatid breaks. These are interpreted as independent breaks on sister chromatids because of preferential effects of the drug in specific chromosome regions. The break points are likely to be different, but morphologically such breaks would be indistinguishable from isochromatid or chromosome breaks.