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Chromosome 15 trisomy in spontaneous and carcinogen-induced murine lymphomas of B-cell origin.

G-banding analyses of 14 independently derived B-cell lymphomas showed the frequent occurrence of chromosome 15 trisomy. It was present in seven of nine spontaneous B-cell lymphomas, but in company with other trisomies, monosomies and marker chromosomes. In five carcinogen-induced primary B-cell leukemias, trisomy 15 was the dominating change. Taken together with the previously demonstrated importance of chromosome 15 trisomy for T-cell leukemogenesis and of the 12;15 translocation in plasmacytogenesis in the mouse, it appears likely that the distal part of chromosome 15 carries a cluster of genes, perhaps a supergene region, that may play an important role in the differentiation and/or the normal responsiveness of various lymphoreticular cell types to growth control.

9,10-Dimethyl-1,2-benzanthracene↗

Cystine uptake by cultured cells originating from dog proximal tubule segments.

Large numbers of kidney epithelial cells were cultured successfully from isolated dog proximal tubule segments. Cells in primary culture and in first passage retained the cystine-dibasic amino acid co-transporter system which is found in vivo and in freshly isolated proximal tubule segments. In contrast to other cultured cells, the cystine-glutamate anti-porter was absent in primary cultures. However, this anti-porter system seemed to be developing in cells in first passage. The intracellular ratio of cysteine:reduced glutathione (CSH:GSH) was maintained at 1:36 in both primary cultures and in low passage cells. Incubation of cells in primary culture for 5 min at 37 degrees C with 0.025 mM [35S]L-cystine resulted in incorporation of approximately 36 and 8.5% of the label into intracellular CSH and GSH, respectively. These cultured cells, therefore, seem to be an excellent model system for the eventual elucidation of a) the inticacies of cystine metabolism and b) regulation of 1) the cystine-dibasic amino acid co-transporter system and 2) the development of the cysteine-glutamate anti-porter system.

Amino Acids↗

T-cell origin of Lennert's lymphoma.

The arrangement of the T-cell receptor and immunoglobulin genes has been analysed in five cases of Lennert's lymphoma. All cases showed rearrangement of the gene coding for the beta chain of the T-cell receptor and a germline configuration of the immunoglobulin genes. This provides strong evidence that Lennert's lymphoma is a T-cell lymphoma.

Antibodies, Monoclonal↗

Secretion of newly taken up ascorbic acid by adrenomedullary chromaffin cells originates from a compartment different from the catecholamine storage vesicle.

Chromaffin cells in primary culture take up [14C]ascorbic acid from the incubation medium. Cells, stimulated immediately after a short labeling period with [14C]ascorbate, secrete ascorbic acid concomitantly with catecholamines (CA) through a nicotinic receptor-mediated Ca2+-dependent process. A proportional release of CA and [14C]ascorbic acid was observed through a large range of secretion rates obtained by varying the concentration of nicotine or by changing the concentrations of Ca2+ and Na+ in the external medium. However, under the same conditions of stimulation, different cell preparations secrete 2-10 times more CA than [14C]ascorbate (as percentage of cell content). Furthermore, a different time course of secretion was observed for CA and [14C]ascorbate for each of several secretagogues. In addition, Ba2+ is a much more potent stimulus for CA secretion than for secretion of [14C]ascorbate, and Ca2+ channel blockers are more potent in inhibiting CA secretion than [14C]ascorbate secretion. These data suggested the possibility that newly taken up ascorbate was being secreted from a compartment altogether distinct from the chromaffin vesicle. This hypothesis was confirmed by subcellular distribution studies, where only a minor fraction of newly taken up [14C]ascorbate was found in the vesicular fraction (P2) from homogenates of chromaffin cells prepared after a short incubation with [14C]ascorbate. However, the subcellular distribution of [14C]ascorbate follows that of endogenous ascorbate when a short pulse with the label is chased by a prolonged equilibration period in the absence of ascorbate, indicating that a transfer has occurred from the extravesicular compartment(s) to the CA storage organelle. Endogenous ascorbate, which is found both inside and outside the chromaffin vesicle, was also found to be secreted from chromaffin cells, indicating that ascorbic acid could be released simultaneously from two different subcellular compartments.

Adrenal Medulla↗

The clinicopathological and prognostic relevance of cytokeratin 7 and 19 expression in hepatocellular carcinoma. A possible progenitor cell origin.

AIMS: Cytokeratin (CK) 7 and CK19 expression, present in hepatic progenitor cells (HPCs) and in cholangiocytes but not in normal hepatocytes, has been reported in some hepatocellular carcinomas (HCCs); however, the incidence and relevance of this expression in HCC in Caucasians is not known. Therefore, our aim was to study the occurrence and clinicopathological characteristics of HCC expressing CK7 and/or CK19 in 109 Caucasian patients. METHODS AND RESULTS: The expression of hepatocellular differentiation markers (Hepar, canalicular polyclonal carcinoembryonic antigen), biliary/progenitor cell markers (CK7, CK19), alpha-fetoprotein (AFP), p53 and beta-catenin in HCC was semiquantitatively assessed by immunohistochemistry. Of 109 HCCs, 78 were CK7-/CK19- (72%), 13 CK7+/CK19- (12%), seven CK7-/CK19+ (6%), 11 CK7+/CK19+ (10%). CK19 expression was significantly associated with elevated serum AFP (400 ng/ml) (P = 0.023), tumour AFP expression (P < 0.0001), presence in serum of anti-hepatitis B core (P = 0.016), less fibrosis in non-neoplastic parenchyma (P = 0.009) and less nuclear beta-catenin expression (P = 0.021). CK7 expression was significantly associated with elevated serum bilirubin (> 2 mg/dl) (P = 0.0005) and less nuclear beta-catenin expression (P = 0.003). HCC expressing CK19 had a higher rate of recurrence (P = 0.009, hazard ratio 12.5, n = 31) after liver transplantation compared with CK19- tumours. CONCLUSIONS: In our series, 28% of HCCs contained cells expressing CK7 and/or CK19. They potentially derive from HPCs. The higher recurrence rate of CK19+ HCC after transplantation suggests a worse prognosis for these HCCs compared with CK19- HCC.

Adult↗

The maturation of reticulocytes. II. Life-span of red cells originating from stress reticulocytes.

Life-spans of macroreticuloytes and macrocytic red cells were studied. Rabbits were made anemic by injecting phenylhydrazine. Peripheral blood rich in reticulocytes was drawn, labeled with 3H-amino acids, and injected back into the anemic animal. Autoradiographic observation on circulating red cells revealed that macroreticulocytes matured at nearly the same time as normal-sized reticulocytes but that the macrocytic red cells had a short life-span compared to normal-sized red cells.

Anemia, Macrocytic↗

Correlation of alkaline phosphatase activity with cell population density in tissue culture cells originated from rat liver.

Alkaline phosphatase (ALP) activity of cultured Li-10 cells obtained from rat liver was found to be a function of cell population density. After the cells grew to confluence, the enzyme activity per cell increased about 100 times that at a low population density. The increase of activity was inhibited by the addition of actinomycin D or cycloheximide to the culture medium. When the cells that had gained high ALP activity after confluency were subcultured, ALP activity decreased to a low basal level after about 48 h. Under cytochemical examination using an electron microscope, the induced ALP activity was seen exclusively at the apical surface region of the cells but scarcely at cell-cell and cell-substratum contact regions.

Alkaline Phosphatase↗

Somite subdomains, muscle cell origins, and the four muscle regulatory factor proteins.

We show by immunohistology that distinct expression patterns of the four muscle regulatory factor (MRF) proteins identify subdomains of mouse somites. Myf-5 and MyoD are, at specific stages, each expressed in both myotome and dermatome cells. Myf-5 expression is initially restricted to dorsal cells in all somites, as is MyoD expression in neck somites. In trunk somites, however, MyoD is initially expressed in ventral cells. Myogenin and MRF4 are restricted to myotome cells, though the MRF4-expressing cells are initially less widely distributed than the myogenin-expressing cells, which are at all stages found throughout the myotome. All somitic myocytes express one or more MRFs. The transiently distinct expression patterns of the four MRF proteins identify dorsal and ventral subdomains of somites, and suggest that skeletal muscle cells in somites originate at multiple sites and via multiple molecular pathways.

Animals↗

Overexpression of ecto-protein kinases in prostasomes of metastatic cell origin.

BACKGROUND: Prostasomes are secretory granules produced, stored, and released by the glandular epithelial cells of the prostate. They express numerous enzymes whose physiological roles have so far not been fully evaluated. In this study, we investigated the expression and function of prostasomal protein kinases and ATPase. METHODS: The protein kinase activities of prostasomes isolated from seminal fluid and malignant prostate cell lines (PC-3, DU145, and LNCaP) were investigated using the model phosphorylation substrates histone and casein, as well as the plasma proteins C3 and fibrinogen, in combination with specific protein kinase inhibitors. The prostasomal ATPase activity was also evaluated. The expression of protein kinases and ATPase on prostasomes was verified by flow cytometry. RESULTS: Prostasomes (intact or solubilized with octylglucoside or saponin) from prostate cancer cells had higher expression of protein kinases A, C, and casein kinase II compared to prostasomes isolated from seminal plasma, resulting in higher phosphorylation of both exogenous and endogenous substrates. Using intact prostasomes, it was found that prostasomes of metastatic origin had lower ATPase activity, resulting in higher residual ATP available for the phosphorylation reaction. Finally, complement component C3 and fibrinogen (two proteins whose activities are modulated by phosphorylation) were identified as physiologically relevant phosphorylation substrates. CONCLUSIONS: These results indicate that prostasomes are capable of modifying proteins possibly involved in the innate response by extracellular phosphorylation mediated by ecto-kinases. This is a novel mechanism by which prostatic malignant cells may interact with their environment.

Adenosine Triphosphatases↗

High-dose therapy with autologous or allogeneic transplantation as salvage therapy for small cleaved cell lymphoma of follicular center cell origin.

Between 1985 and 1996, 51 patients with relapsed or refractory small cleaved cell lymphoma (SCCL) received high-dose chemotherapy +/- TBI in conjunction with autologous (ABMT) (36 patients) or allogeneic transplantation (15 patients). Patients were eligible for ABMT if the bone marrow biopsy done prior to the planned transplant did not reveal microscopic involvement with SCCL. Patients receiving ABMT had a median age of 48 years, had received a median of 2.5 chemotherapy regimens prior to transplantation, and were transplanted a median of 35.5 months from diagnosis. Among patients receiving ABMT, 5 year actuarial survival was 56+/-11%. Median survival was 126+ months, and median survival from diagnosis was 191 months. Univariate and multivariate analysis identified sensitive disease as the best predictor of a favorable response. Five-year actuarial survival was 66+/-12% for patients with sensitive disease at the time of transplant as compared to 29+/-17% for patients with resistant disease, P = 0.015. Median survival in patients with sensitive disease at the time of ABMT was 126+ months. By univariate analysis, survival was significantly better for patients receiving ABMT as compared to patients receiving allogeneic transplants. Median survival following allogeneic transplantation was 5 months; 5 year actuarial survival was 15+/-13%. In a multivariate analysis, which considered autologous vs allogeneic transplantation, sensitive vs resistant disease, <3 vs > or = 3 prior treatments, and prior bone marrow involvement, allogeneic transplantation was significantly associated with poor survival. Treatment-related mortality occurred in eight of 15 patients receiving allogeneic transplantation and limited the effectiveness of this therapy. High-dose therapy in conjunction with ABMT is effective therapy for patients with SCCL whose disease is sensitive to chemotherapy and whose marrows are microscopically free of disease. Because of possible selection bias, it has not been proven that this approach increases survival in these patients. Treatment-related mortality limits the effectiveness of allogeneic transplantation in SCCL.

Adult↗

Ex vivo induction of anti-leukemia cytotoxic T cell effect by dendritic cells from human umbilical cord blood cell origin.

To explore the possibility of in vitro induction of cord blood cell-derived lymphocytes into cytotoxic T lymphocytes (CTL) with anti-leukemia specificity, umbilical cord blood (UCB)-derived mononuclear cells were cultured with multiple cytokines to generate dendritic cells (DC) in vitro. Leukemia cells were irradiated with (137)Cs and activated by premature cytokines. The characteristics of maturation of DC were evaluated through morphology examination and flow cytometry. DC pulsed with leukemic antigens were co-cultured with lymphocytes. Cytotoxicity of the CTL to corresponding leukemic cells was measured with lactate dehydrogenase-release assay. The results showed that UCB-derived monocytes could be induced into typical DC in all of the 12 samples. Expression of immunological markers such as CD1a(+), HLA-DR(+), CD86(+), CD83(+) on DC were significantly up-regulated (P < 0.05). DC presenting leukemic antigens generated leukemia-specific CTL with a killing rate of (44.76 +/- 17.42)% at the E:T ratio of 50:1 against AML cells and a killing rate of (8.50 +/- 4.25)% at the E:T ratio of 50:1 against ALL cells. Whereas, these CTL present almost no killing effect on the mononuclear cells collected from the same patients in complete remission phase. It is concluded that (1) it is possible to induce UCB-derived monocytes into mature DC with typical morphology. (2) Cord blood derived mature DC presenting leukemia antigen can generate leukemia-specific CTL with vigorous cytotoxic activity against the same leukemia blasts and low killing activity against bone marrow cells of the same patients in complete remission phase.

Cells, Cultured↗

Emergence of a B-cell lymphoblastic lymphoma in a patient with B-cell chronic lymphocytic leukemia: evidence for the single-cell origin of the two tumors.

A patient is described who presented with a chronic lymphocytic leukemia (CLL) and later developed a lymphoblastic lymphoma. The cells from the CLL were typical mature B lymphocytes as could be assessed by morphologic, cytochemical, and surface marker analyses. The cells from the lymphoblastic lymphoma were immature B cells that expressed CD10, CD20, and HLA-DR markers, but not surface Ig or cytoplasmic mu chains, and were negative for terminal deoxynucleotidyl transferase (TdT). The cells of two continuous cell lines, obtained from the bone marrow and the peripheral blood of the patient, had the same phenotype as the lymphoblastic lymphoma cells, did not contain the Epstein-Barr virus genome, and displayed malignant features in vitro, including the capacity to form colonies in agar. The two cell lines also shared identical chromosomal abnormalities, a finding which suggests that they derived from the same malignant cell already present in vivo. Such chromosomal abnormalities were not seen in the karyotype of the peripheral blood cells at the onset of the disease. Analysis of the Ig heavy chain genes using a DJ-specific probe showed the very same monoclonal rearrangement in the cells from the B-CLL, the lymphoblastic lymphoma and the two cell lines, thus demonstrating their common clonal origin. By contrast, a monoclonal rearrangement of the lambda chain gene locus was found in the B-CLL cells only, a finding consistent with their exclusive capacity to express surface IgM lambda. This patient represents a rare case in whom a chronic lymphoproliferative disorder with mature malignant cells transforms into a lymphoblastic lymphoma characterized by cells frozen at a very early maturational stage. The possible mechanisms leading to such transformation within the same cell clone are discussed.

Adult↗

[Effect of oxLDL on the uptake and clearance rate of cholesterol in vascular smooth muscle cells originated from human apoAI transgenic mice].

OBJECTIVE: Study on (1) inhibition of oxidized low density lipoprotein (oxLDL) effect on the uptake and clearance of intra-cellular 3H-cholesterol in vascular smooth muscle cells (v-SMC) originated from the human-apoAI transgenic mice (C57BL/6); (2) change of human-apolipoprotein AI (h-apoAI) mRNA expression in v-SMC after oxLDL stimulation and the protective effect of expressed h-apoAI on v-SMC against oxLDL intoxication. METHODS: (1) v-SMC isolated from human apoAI transgenic mice possessing a recombined gene connected beforehand with a mouse metallothionein-I (MT-I) as the promoter; (2) study of h-apoAI mRNA expression from v-SMC of the transgenic mice by RT-PCR and Northern blot. RESULTS: oxLDL (30 micrograms/ml) strongly promoted v-SMC proliferation. No difference found on 3H-cholesterol uptake between smooth muscle from normal mouse aorta (n-SMC) and smooth muscle cells from transgenic mouse aorta (tr-SMC) of the control groups, the uptake rates of both kinds of SMC rose 100% after oxLDL stimulation. The efflux rates of 3H-cholesterol in tr-SMC were much higher than those of n-SMC (40%-50%). After oxLDL stimulation, the clearance rates fell by 28% and 10% respectively for n-SMC and tr-SMC. The result of RT-PCR and Northern blot showed a marked increase of h-apoAI gene expression after oxLDL stimulation. CONCLUSIONS: Expression of h-apoAI gene in C57BL/6 mice enables to decrease the accumulation of cholesterol in v-SMC. tr-SMC are capable to alleviate the harmful effect of oxLDL on v-SMC due to the increase of h-apoAI expression.

Animals↗

Stem cell origins of leukaemia and curability.

It is suggested that most childhood acute lymphoblastic leukaemias and some other paediatric cancers are chemo-curable because they arise in stem cell populations that are functionally transient, chemosensitive and programmed for apoptosis. Most adult acute leukaemias are chemo-incurable at least in part because they originate in relatively drug resistant stem cells with extensive self-renewal capacity. The latter property in turn increases the probability of clones evolving with multi-drug resistance. Particular mutations may superimpose additional adverse features on leukaemic cells.

Adult↗

Cytogenetic studies of a diffuse mixed cell lymphoma of T cell origin.

The chromosome finding obtained from a lymph node of a patient with T cell lymphoma is described. Two different abnormal clones were found; one of them had a t(14;18)(q32;q21), along with other structural and numerical abnormalities, including 1p+q-,1p-,2p+q+, der(11),t(11;?)(q13;?),+20,+21,22p+. The other clone contained der(13),t(13;?)(q22;?).

Adolescent↗