Assay of effect of (2'-5')-oligoadenylate on macrophages.
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Biomedical subjects
Publications and source records attributed to N Wang.
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A patient presented with a primary melanoma, Level IV, 2.5 mm thick; 30 excised lymph nodes were all negative for tumor. Four local recurrences followed in the ensuing 17 months. Tumor cells cultured at this time were epithelioid. Autoimmunization was followed by a disease-free interval of 15 months. Postimmunization, the patient's lymphocytes destroyed his melanoma cells in culture and were stimulated in mixed cell culture by his irradiated tumor cells. Cells grown from the relapsing tumor were spindle/dendritic with bizarre morphology and were not attacked by his lymphocytes in culture. Using a C' fixation technique, DR antigen profiles of the patient's B-cells and both tumor cell types showed that the immunizing tumor was positive for DR antigens 3, 5, and 8, none of which were present on his B-cells which had DR 2 and 4. Both B-cells and immunizing tumor cells were positive for DQ antigens. The relapsing tumor cells were positive for DR2 and negative for all other D region antigens tested. The evidence suggests that given a melanoma of heterogeneous cell population, autoimmunization against the predominant immunogenic cell inhibits tumor growth but allows the ascendance of a nonimmunogenic tumor cell type.
It is paradoxical that the inactivated X is the only chromosome that can be identified in the interphase nucleus, yet in metaphase, it is indistinguishable from its genetically active homolog unless special culture and staining procedures are employed. A specific inactivation-associated fold in proximal Xq resolves that paradox. We describe here how the fold in the proximal long arm can be used as a simple and reliable marker to identify the inactivated X in G-, Q-, or R-banded preparations. Several examples are given, including localization of the inactivation center to band Xq13 or q21.1, identification of nonrandom inactivation in X-chromosome rearrangements, identification of multiple active X chromosomes in tumor cell lines, analysis of X-inactivation patterns in female carriers of the fragile site at Xq27, and comparison of X-inactivation patterns among primate species.
Sister chromatid exchanges (SCE) were studied in lymphocyte cultures of 12 adult male epileptic patients on long-term monotherapy with phenytoin (PHT) and of matched controls. Significantly increased frequency of SCE was observed in the epileptic patients as a group and in almost all individuals, indicating a detectable chromosome damaging effect of PHT therapy on its human users.
Two prepubertal sisters of American Indian origin developed osteosarcoma at 8 and 12 years of age. This familial occurrence, tumor onset prior to puberty, unusual tumor location in one who also had short stature, and ethnic background all suggest an inborn predisposition to bone cancer rather than a chance occurrence. Rearrangements involving chromosomes #13 and #14 were found in both the surviving proband and mother. Comparison of the arm ratio and prometaphase G-banding patterns of the rearranged chromosomes suggests either deletion of band 14q11.2 or pericentric inversion (with breakpoints at 13q12 and 14q11.2) in the proband's rearranged chromosome, but not in her mother's. Her mother, who had no malignancy, had a typical Robertsonian translocation [t(13;14)(p11;q11)]. Three previously reported children with chromosomal abnormalities developed osteosarcoma at unusually young ages, younger even than in reported sibships with osteosarcoma. The most frequently detected cytogenetic abnormalities in sarcoma tumor cells involve chromosomes #13 and #14. In addition, some cases of bilateral retinoblastoma and familial unilateral retinoblastoma, which are known to be at increased risk for osteosarcoma, are associated with tiny deletions on chromosome #13. Thus, there may be a causal relationship between constitutional loss or rearrangement of genetic material at these breakpoints on chromosomes #13 or #14 and development of osteosarcoma in this family that is similar to that seen in patients with small constitutional chromosomal deletions who develop Wilms' tumor and retinoblastoma.
It has recently been shown that 5-azacytidine (5-azaC) can induce altered replication patterns of the late-replicating X chromosome in normal female cells. This has been demonstrated by bromodeoxyuridine labelling of cells late in the S phase. In the present study the same method was applied to the lymphocytes of a Klinefelter patient (48,XXXY/47,XXY). Significant 5-azaC-induced changes in the replication of the entire inactive X chromosome, from late to early, were found in the lymphocytes of this patient. These results indicate that hypomethylating agents can not only alter the replication of individual bands, but also change the gross replication schedule of multiple inactive X chromosomes in the presence of a Y chromosome.
The amino acid sequence of the precursor to the second-most-abundant serum antifreeze protein (B) in the winter flounder has been determined by a combination of protein and DNA sequencing. The precursor is an 82-residue preproprotein which differs in only three positions from the amino acid sequence of the precursor for the most-abundant serum antifreeze protein (A). The base changes responsible for these substitutions, as well as several silent changes, are all clustered within the DNA coding for the mature protein portion. Among the post-translational modifications that the precursor undergoes is the removal of the c-terminal glycine residue. Cloning of full-length antifreeze protein cDNA has enabled us to identify the transcription initiation site which occurs 49 nucleotides upstream from the initiation codon.
A 3-Methylcholanthrene (MC)-induced fibrosarcoma and three of its clones were investigated for their metastatic potential in normal and tumor immune mice. The growth rates of the four tumors in vivo were similar. However, the mean survival times of the tumor-bearing mice were markedly different. Clone 10, the most immunogenic, showed very high metastatic potential and short survival, while clone 27, the least immunogenic, produced few metastases, resulting in much longer survival. Moderate numbers of metastases were produced by highly immunogenic 3-AM (parental tumor), and poorly immunogenic clone 34. Spleen cells from mice bearing highly immunogenic tumors lost their ability to neutralize tumors by day 28 after tumor inoculation, while those from mice bearing poorly immunogenic tumors remained cytotoxic, indicating that highly immunogenic tumors also induced immune suppression in the hosts. Immunization with specific tumors decreased the number of pulmonary metastases by 3 to 35-fold. Immunization with tumors that shared antigens provided protection against metastatic tumors as well as the local tumors. In contrast, immunization with antigenically different tumors gave no protection.
High resolution prometaphase G-banding analysis was applied to three translocation carriers from the t(3;8) hereditary renal cell carcinoma family. It was clearly illustrated that the chromosomal rearrangement is reciprocal with breakpoints occurring at the subbands 3p14.2 (instead of 3p21) and 8q24.1.
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pppA2'p5'A2'p5'A(2'-5'P3A3) activates macrophages and increases the phagocytosis of macrophages from different species including human beings. This indicates that the activation of macrophages may be a general action of 2'-5'P3A3. This discovery broadens the effect of 2'-5'P3A3 beyond the antiviral field. alpha-Feto-protein (AFP) inhibits the phagocytosis of macrophages and may be involved in the development of hepatoma. Data presented here show that 2'-5'P3A3 can antagonize this suppressive effect of AFP. Methods used so far for introducing 2'-5'P3A3 into the cells were made with the aid of CaCl2, etc. under conditions which may not be the same as those used clinically. It was found that 2'-5'P3A3 can develop its biological effect without the aid of CaCl2, etc.
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Two formaldehyde-induced, homozygous viable ADH-negative mutants, Adhfn4 and Adhfn6, possess no material that cross-reacts with antibody directed against ADH, no mature mRNA of wild-type size, and greatly reduced amounts of RNA that hybridizes with an Adh probe. We have cloned the genomic DNA sequences from these mutants in bacteriophage lambda Charon 4 and subcloned the Adh region into plasmid vector pBR327. Restriction analyses revealed one small deletion in each of these mutants and DNA sequencing showed that the splice junctions of the 65-base pair (bp) intervening sequence (IVS) were altered. Both cloned mutant Adh genes, as well as the wild-type gene, are capable of promoting correct specific transcription initiation in HeLa cell nuclear extracts in vitro. We conclude that Adhfn4 and Adhfn6 are defective in RNA processing. Our results provide evidence for the importance of the splice junction sequences in normal ADH RNA processing and stabilization in Drosophila. We also speculate that splicing of ADH RNA proceeds in a nonrandom manner: mutations in one of the intervening sequences appear to cause accumulation of a large ADH RNA containing at least one other IVS.
Biological characteristics were assessed in GR mouse mammary tumors during 22 serial transplantations. Although unidirectional progression from hormone dependency to independency was observed, other biological markers such as progesterone receptors, polyploid frequency and thymidine kinase activity demonstrated cyclic phenomena every fourth to sixth transplant generation, suggesting the continued presence of regulatory mechanisms among various cells subpopulations.
Regressing atypical histiocytosis (RAH) of skin is a cutaneous noduloulcerative proliferation of atypical neoplastic histiocytes with concomitant polymorphous inflammation, frequently pronounced epidermal hyperplasia, and an unexpectedly indolent biologic course. Spontaneous regression and recurrence without systemic spread were the course in follow-up periods of over six years. Histopathologically, characteristic-appearing atypical mononuclear and multinucleated "RAH" cells showed erythrophagocytosis as well as ultrastructural, surface marker, and enzyme cytochemical features indicating histiocytic differentiation. Cytogenetic analysis showed aneuploidy and several marker chromosomes including 14q+. Its benign biologic course clearly distinguished this entity from malignant histiocytosis, large cell lymphoma, and Hodgkin's disease. The histiocytic atypical cells further distinguished it from the T-cell lesions of the skin, such as mycosis fungoides and lymphomatoid papulosis. This entity is readily confused with malignant lymphoreticular disease, melanoma, or squamous carcinoma.