Guidelines for the clinical diagnosis of Lyme disease.
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False Negative Reactions↗
Biomedical subjects
J Kochevar
Publications and source records attributed to J Kochevar.
Isolation of brefeldin A.
1. A chromatographic method for the isolation of brefeldin A (BFA) from the culture medium of Penicillium brefeldianis has been developed. BFA is adsorbed on a reversed-phase (C18) silica, desorbed by methanol, and the crystalline BFA precipitated from the concentrated aqueous solution. The yield of crystalline BFA is 121 mg/l of culture medium from a sample containing 204 mg/l determined by analytical HPLC. 2. The detection limit of BFA is 50 ng by analytical HPLC and BFA in the mother liquor of crystallization is recovered by preparative HPLC.
Anti-Bacterial Agents↗
Blockage of autonomous growth of ACHN cells by anti-renal cell carcinoma monoclonal antibody 5F4.
A cultured renal carcinoma cell line, ACHN, continued to proliferate in the absence of exogenous growth factors supplied by fetal calf serum. ACHN cells had been previously used as the immunogen for the production of monoclonal antibody 5F4. Subsequent immunohistochemical studies showed that 5F4 strongly and selectively reacted with renal carcinoma cells compared with normal renal epithelium, suggesting that expression of the substance to which it was bound was amplified as a result of malignant transformation. Because renal carcinoma cells are relatively differentiated with regard to function except for elements to ensure sustained growth, the immunoreactivity patterns led to the hypothesis that 5F4 reacted with some component of a growth-stimulating pathway. The hypothesis was supported in the present work by showing that 5F4 specifically inhibited the growth of ACHN cells; ACHN cells prominently secreted a protein with a molecular mass of 178 kilodaltons that was immunoreactive with 5F4; the protein was partially purified by simple lyophilization of serum-free conditioned medium; both ACHN conditioned medium and the lyophilizate stimulated the growth of quiescent human fibroblasts and BALB/3T3 cells as well as serum-deprived human renal carcinoma cell lines ACHN, A498, Caki-1, and Caki-2. Specific immunoadsorption of the protein by affinity chromatography with 5F4 removed the growth-stimulating activity. The results demonstrated a novel growth-promoting substance secreted by ACHN cells with autologous activity as well as activity for human and murine fibroblastic cell lines and other renal carcinoma cell lines. The growth-promoting activities were specifically blocked by monoclonal antibody 5F4.
Animals↗
Low level amplification of c-sis and c-myc in a spontaneous osteosarcoma model.
Canine and human osteosarcoma are very similar clinically, radiologically and pathologically. DNA extracted from canine osteosarcomas (n = 9) and normal canine control tissues (n = 17) was examined for amplification of the c-sis, c-myc, N-myc and c-H-ras protooncogenes. Statistically significant amplification of the c-sis and c-myc protooncogenes was evident in the tumor tissues as compared to the normal control tissues (P less than 0.05). DNA and total cellular RNA from cultured canine and human osteosarcoma and fibroblast cell lines were examined for amplification or enhanced expression of c-sis and c-myc. Very low levels of c-myc and c-sis DNA amplification were noted in canine osteosarcoma cells as compared to canine fibroblasts. Immunostaining of sections of human and canine osteosarcoma for the sis gene product, PDGF B, showed similar levels and patterns of expression in both populations of tumors.
Animals↗
Differential immunostaining of oncocytic renal tumors with an anti-renal cell carcinoma monoclonal antibody.
Formalin-fixed, paraffin embedded tissue sections from twenty three oncocytic renal neoplasms were stained with hematoxylin and eosin, retrospectively examined, and graded according to the criteria reported by Lieber and associates. Additional sections were stained by the avidin-biotin immunoperoxidase technique with anti-renal cell carcinoma monoclonal antibody 5F4. The results showed that cytological heterogeneity was the most prominent feature of the tumors. Four cases were composed predominantly of grade 1 cells, but also had foci of grade 2 cells. Seventeen cases were composed predominantly of grade 2 cells. Two cases were composed predominantly of grade 3 cells. Immunostaining with 5F4 showed differential reactivity between grade 1 and grades 2 and 3 cells. The antibody highlighted the foci of atypical cells which were difficult to detect by routine hematoxylin and eosin staining and thus could be useful in the differential diagnosis of oncocytic renal neoplasms.
Adenoma↗
A renal cell carcinoma neoplastic antigen detectable by immunohistochemistry is defined by a murine monoclonal antibody.
BALB/c mice were hyperimmunized with ACHN (ATCC CRL 1611, American Type Culture Collection, Rockville, Maryland), a stable in vitro cell line derived from a malignant pleural effusion in a 22-year-old man with renal cell carcinoma. The hyperimmune spleen cells were fused with NS-1 murine myeloma cells using polyethylene glycol. Hybridoma supernatants were screened for the presence of IgG reactive with detergent extracts of ACHN and nonreactive with detergent extracts of normal kidney tissue. A stable, rapidly growing clone named 5F4 was isolated. Supernatant from 5F4 was used as a primary antibody preparation for avidin-biotin complex immunoperoxidase staining of multiple cases of renal cell carcinoma, normal tissues, and other tumors. 5F4 produced IgG which reacted with a cytoplasmic structure in paraffin-embedded sections of all renal cell carcinomas tested. There was occasional, weak, granular, cytoplasmic staining of isolated tubular lining cells in adjacent normal kidney.
Animals↗
Adenocarcinoid tumor, goblet cell type, arising in a ureteroileal conduit: a case report.
The adenocarcinoid tumor is a potentially malignant neoplasm usually arising in the appendix. We describe a 55-year-old woman with a ureteroileal conduit in place for 12 years who suffered obstruction of the ileal segment of the conduit, with recurrent pyelonephritis and abdominal pain. Pathological examination of a segmentally resected portion of the conduit revealed a submucosal proliferation of nests of argentaffin/argyrophil cells and large, mucin-distended goblet cells. The patient showed no evidence of metastasis and has done well to date following excision. A discussion of neoplasms arising in the ureteroileal conduit is included.
Carcinoid Tumor↗
Perceived Somatic Response Inventory: three scales developed by factor analysis.
Studies of autonomic reactivity and of voluntary control of autonomic responses have suggested the importance of individual differences. An inventory of scales was developed to improve the assessment of different dimensions of somatic functioning. In order to enhance their specificity and incremental validity, the scales were constructed to be relatively independent of already known general personality variables such as Stress Reaction (neuroticism), Locus of Control, Hypochondriasis, and Absorption. Three scales, derived from orthogonal factor analyses, were developed and cross-validated: (1) Autonomic Response Frequency, assessing spontaneous fluctuations in autonomic responding; (2) Autonomic Response to Stress, assessing the amount of autonomic arousal perceived by a subject in stress situations; and (3) Somatic Response Control, a self-report measure of capacity to control both autonomic and fine muscle responses. The hypothesis that sympathetic and parasympathetic awareness might define two distinct dimensions was not supported by the data. The new scales will be useful in testing hypotheses of possible relationships among the factors measured and behavioral and psychophysiological indicators of the response to stress.
Adult↗