Search PubMed⌕ Search

Biomedical subjects

J A Roth

Publications and source records attributed to J A Roth.

At least 469 records · Page 26Linked to original sources
PubMed1984

A novel approach to production of antitumor monoclonal antibodies: antibody to a cell surface glycoprotein associated with transformation by a human oncogene.

Transfection is a technique for inducing transformation of normal fibroblasts (NIH 3T3) with DNA (oncogenes) from human tumors. Our goal was to determine if these transformed cells expressed antigens associated with malignancy. NIH 3T3 cells were transfected with DNA fragments from a human acute lymphocytic leukemia (ALL 1-69), and transformed colonies were selected for growth in soft agar. Transfected cells containing human DNA sequences demonstrated by Southern blot analysis were used to immunize Balb/C mice. Monoclonal antibodies were produced and screened for binding to the parental leukemia (ALL 1-69), transfectant (17(2], and 3T3 cells in an enzyme-linked assay. A monoclonal antibody (IgM kappa) designated 17-9H3 bound to ALL 1-69 and secondary transfectant 17(2) but not to NIH 3T3 plasma membranes. Immunoperoxidase staining confirmed this binding pattern and demonstrated that the antigen was expressed on the cell surface. Expression of the antigen by transfectants directly correlated with the presence of a single 6.1 kilobase human DNA sequence. The antibody binding site of the antigen was inactivated by trypsin, glucosidase, and hyaluronidase. Binding of the 17-9H3 antibody was selective for acute lymphocytic leukemias (5/8) and osteogenic sarcomas (33/36), although other tumor types did demonstrate significant binding by immunoperoxidase staining. The majority of normal tissues did not bind 17-9H3 with the exception of some metabolically active cells (renal tubular epithelium, secretory epithelial cells, and cardiac smooth muscle), germ cells, Leydig cells of the testes, and some lymphoid cells. Monoclonal antibodies to oncogene-associated antigens may be potentially useful for cancer diagnosis and therapy and as probes for oncogene isolation.

Animals↗
PubMed1984

Analysis of antigenic expression by primary and autologous metastatic human sarcomas using murine monoclonal antibodies.

The efficacy of monoclonal antibody therapy depends in part on the expression of the relevant tumor-associated antigens by both primary tumors and their metastases. Antigen expression by paired primary and autologous metastases from surgically excised osteogenic and soft-tissue sarcomas from 15 patients was studied using a panel of murine hybridoma monoclonal antibodies and indirect immunoperoxidase staining of formalin-fixed tissue sections. The panel included three antibodies (B3619, 17-9H3, OST6) recognizing sarcoma-associated antigens and an antibody recognizing an HLA-DR framework determinant (OKla1). In most cases, antibody binding to both primary and metastatic tumors was observed. However, marked heterogeneity of binding intensity between primary and metastatic tumors and of cells expressing antigens within tumors was noted. This occurred even though primary and metastatic tumors demonstrated homogeneous histology and cellular morphology. Differences were noted among patients as well as among metastases taken from an individual. A significant number of both primary and metastatic tumors contained cells that did not bind a particular antibody even in the presence of other cells that demonstrated significant antibody binding. Thus, strategies for single monoclonal antibody therapy may be limited by heterogeneity of intertumor and intratumor antigenic expression.

Animals↗
PubMed1984

Pilot study on the treatment with thioproline of 24 small animals with tumors.

Thioproline (thiazolidine-4-carboxylic acid) was administered in a 16-week trial by either IM or subcutaneous routes to 23 dogs and 1 cat with solid tumors (12 animals) and incompletely excised malignant tumors (12 animals). Acute partial responses were observed in 3 of the 12 animals with solid tumors. In a mammary gland carcinoma and oral fibrosarcoma, there was tumor enlargement by the 4th day of drug therapy followed by partial regression. Drug activity appeared to be directed against tumor blood supply. Tumor progression occurred in 11 of the 12 other animals (incompletely excised tumors). Although 4 of 5 dogs with intranasal tumors were alive and relatively free of tumorous signs at 9 months or more after surgical operation was done, there was no microscopic evidence of reverse transformation or cytostatic activity in the 2 intranasal tumors that were reoperated upon. There was no hematologic or biochemical toxicosis noted. Apparent pain from injection and abscess formation were the most frequent complications from parenteral administration and would preclude using this method on a long-term basis.

Animals↗
PubMed1984

Equine cell-mediated immune response to Rhodococcus (Corynebacterium) equi.

A lymphocyte blastogenic assay was developed to serve as an in vitro correlate of cell-mediated immunity to Rhodococcus (Corynebacterium) equi (R equi) in the equine species. Lymphocytes obtained from a group of experimental ponies showed no response in cell culture to R equi heat extract or lysozyme extract antigens. Ponies were assigned to groups for experimental inoculation. Three ponies were inoculated subcutaneously with live R equi, 3 were given live R equi by intranasal and intratracheal routes, and 4 ponies were left untreated. Lymphocytes from all inoculated ponies had a mitogenic response to R equi antigens in lymphocyte blastogenic assays performed between the 7th and 40th days after inoculation. Lymphocytes from noninoculated control ponies remained unresponsive to R equi antigens. Delayed-type hypersensitivity reactions developed in all experimentally exposed ponies after intradermal administration of the R equi antigen preparations. In a 2nd phase of experimentation, blastogenesis assays were performed on lymphocytes from horses in herds with endemic R equi infections. Results indicated that many of the animals had significant (stimulation index greater than 2) cell-mediated responses to the bacterium, but there was no distinct correlation between the immune response and clinical history. These data indicated that cell-mediated immunity is involved in the interaction of the equine immune system with R equi.

Animals↗
PubMed1984

Equine humoral immune response to Rhodococcus (Corynebacterium) equi.

An enzyme-linked immunosorbent assay was developed to test equine serum for the presence of antibodies to Rhodococcus (Corynebacterium) equi. Experimental ponies had no detectable antibody to R equi before exposure to the bacterium. After experimental inoculation, animals in groups that received live R equi subcutaneously or intranasally/intratracheally developed high titers to R equi. Noninoculated controls remained seronegative. Serum was also collected from horses of various ages that were naturally exposed to R equi. There was a wide range of anti-R equi titers in these horses. Because experimentally infected horses seroconverted when some naturally infected foals did not seroconvert, the function of antibody in resistance to R equi infection remains unknown.

Animals↗
PubMed1984

Analysis of prognostic factors in patients undergoing resection of pulmonary metastases from soft tissue sarcomas.

The selection of objective criteria that can reliably predict survival in patients undergoing resection of pulmonary metastases remains controversial. Between 1974 and 1982, 487 patients with soft tissue sarcomas presented to the National Cancer Institute. Eighty patients underwent thoracotomy for putative metastases and 67 patients had histologically proved pulmonary metastases. The 3 year tumor-free survival rate was 30% by actuarial analysis. Patients with resectable metastases had significantly prolonged post-thoracotomy survival compared to those patients with unresectable metastases. The most significant preoperative predictors of survival were the tumor doubling time, the number of metastases on preoperative linear chest tomograms, and the disease-free interval. Patients with a tumor doubling time of 20 days or more had a significantly longer post-thoracotomy survival (22 months median) than patients with a tumor doubling time of less than 20 days (6 months median). Those patients with four nodules or less on preoperative tomograms had significantly longer post-thoracotomy survival times (23 months median) than those patients with more than four nodules (6 months median). Patients with a disease-free interval of more than 12 months had a longer post-thoracotomy survival (32 months median) than patients with a disease-free interval of 12 months or less (10 months median). Combining these three prognostic factors significantly increased the predictive ability of this model. These criteria provide an accurate and rapid method to identify preoperatively those patients who will maximally benefit from resection of pulmonary metastases from soft tissue sarcomas.

Adolescent↗
PubMed1984

Functional and ultrastructural evaluation of neutrophils from foals and lactating and nonlactating mares.

Neutrophils from 4 pony foals, 3 lactating pony mares, and 3 nonlactating mares were evaluated ultrastructurally and by in vitro function tests. Neutrophils from foals had significantly (P = 0.05) less random migration than neutrophils from mares; values in tests for iodination and Staphylococcus aureus ingestion were also lower with foal neutrophils. Neutrophils from lactating mares had lower responses to iodination, antibody-dependent cell-mediated cytotoxicity, and random migration tests than did neutrophils from nonlactating mares. Ultrastructurally, granule concentration did not differ significantly among groups. A slight decrease in primary granules and a corresponding increase in granules with a flocculent matrix indicates partial spontaneous neutrophil degranulation in foals and lactating mares.

Animals↗
PubMed1984

Improvement in clinical condition and thymus morphologic features associated with growth hormone treatment of immunodeficient dwarf dogs.

Immunodeficient dwarfism in Weimaraner dogs was characterized by failure to grow, emaciation, growth hormone (GH) deficiency, decreased lymphocyte blastogenic responsiveness to mitogens, lack of thymus cortex, and recurrent infections usually resulting in death. Affected pups did not respond to conventional supportive therapy, but did respond to treatment with thymosin fraction 5. Response to therapy with bovine GH was monitored by clinical observation, histopathologic examination of thymic biopsy material, lymphocyte blastogenic responsiveness to nonspecific mitogens, and radioimmunoassay of thymosin alpha 1 concentration in the serum. Growth hormone therapy (0.1 mg/kg of body weight/dose, 14 doses) during a 1-month period in 2 immunodeficient dwarf pups resulted in clinical improvement and a marked increase in the thickness and cellularity of the cortex of the thymus. Immunodeficient dwarf pups were not deficient in serum thymosin alpha 1 before GH therapy. Growth hormone therapy was not associated with a consistent increase in serum thymosin alpha 1 concentration or lymphocyte blastogenic responsiveness to mitogens.

Animals↗
PubMed1983

Lymphokine-activated killer cell phenomenon. III. Evidence that IL-2 is sufficient for direct activation of peripheral blood lymphocytes into lymphokine-activated killer cells.

Purified interleukin 2 (IL-2) was found to be sufficient for direct activation of peripheral blood lymphocytes into lymphokine-activated killer (LAK) cells. The LAK activation factor was directly and consistently associated with IL-2 activity using classic protein purification techniques, adsorption to IL-2-dependent cell lines, and inhibition with anti-Tac antibody. As yet, no other cytokines have been found that perform the same role.

Antibodies, Monoclonal↗
PubMed1983

Human brain monoamine oxidase: solubilization and kinetics of inhibition by octylglucoside.

Complete solubilization of both the A and B forms of human brain monoamine oxidase (MAO) occurred when crude mitochondria were incubated in the presence of 50 mM octylglucoside (OG). Upon removal of this nonionic detergent by dialysis, approximately 100% of the starting activity was present in the dialysate. The effects of solubilization were examined by comparison of several properties of the membrane-bound and OG-treated oxidases. The percentage inhibition of phenylethylamine (PEA) and the 5-hydroxytryptamine (5-HT) deamination by deprenyl and clorgyline were identical. The Km values obtained for the deamination of PEA, a B-selective substrate, 5-HT, an A-selective substrate, and tyramine (TYR), a nonselective substrate, were also comparable. OG was found to inhibit type A (I50 = 8.1 mM) and B (I50 = 4.7 mM) MAO activities at concentrations at least 10-fold below those used to solubilize the oxidases. Kinetic studies revealed that OG was an apparent competitive inhibitor of PEA deamination whereas OG produced a mixed-type pattern of inhibition when 5-HT was the variable substrate. Inhibition of TYR deamination by either the A or B form of MAO produced a mixed pattern of inhibition. The findings herein suggest that solubilization of the A and B forms of MAO by OG does not significantly alter the substrate and inhibitor specificity of the oxidases following removal of detergent. However, in the presence of concentrations of OG 50 times less than the critical micellar concentration of this detergent, marked inhibition of deamination by both forms of human brain MAO is observed. Accordingly, the usefulness of OG is limited to situations where the detergent is completely removed before quantitation of MAO activity.

Brain↗
PubMed1983

Activity of neuron-specific enolase in normal and lesioned rat brain.

Rat brain contains 3 forms of enolase, a neuron-specific form (NSE), a hybrid form, and a non-neuronal form (NNE) which were separated by DEAE-cellulose column chromatography. The enolase activity corresponding to each form of the enzyme eluted from the columns was determined spectrophotometrically. Using this assay procedure, an activity ratio (%NNE/%NSE) was calculated for cerebellum, brainstem, sciatic nerve, adrenals and liver. The results indicated excellent agreement between this enzymatically determined ratio and published values of a similar ratio (NNE/NSE) determined by radioimmunoassay for enzyme protein. Following in vivo destruction of neurons by intracerebral injection of the selective neurotoxin, kainic acid, there was a significant decrease in the activity of NSE and hybrid enolase (neuronal forms) and no change in the activity of NNE (glial form). These data indicate that separation and measurement of enolase species is useful to determine levels of these species in normal tissue and to estimate neuronal damage biochemically in brain lesions.

Animals↗
PubMed1983

Detection of a suppressive immunoregulatory factor (IRF) in the sera of sarcoma patients by enzyme-linked immunoassay (ELISA) and correlation with clinical course.

Immunoregulatory factor (IRF) is a 70,000 molecular weight glycoprotein produced by human tumors that suppresses lymphocyte function including mitogen-stimulated tritiated leucine (3H-leu) and tritiated thymidine (3H-Tdr) uptake, in vitro immunoglobulin synthesis, induction of allospecific cell-mediated cytotoxicity, and proliferation of T-cell growth factor-dependent lymphocyte cultures. Antisera to IRF were produced by immunization of goats and rabbits with IRF purified by diethylaminoethyl (DEAE)-anion exchange and affinity chromatography. Antisera specificity was demonstrated by binding to IRF but not control muscle extract in an ELISA test and by specific removal of IRF activity by antibody coupled to acrylamide beads. Using these antisera, a double antibody-binding assay was developed to measure circulating levels of IRF and to determine its relationship to tumor growth and relevance for monitoring the clinical course of cancer patients. Quantitative autologous as well as allogeneic binding of sarcoma patients' sera to anti-IRF antibody was demonstrated. Of five tumor extracts tested, two had immunosuppressive activity, and IRF was detected in the preoperative sera of both patients. These extracts were not suppressive and IRF was not detected in the preoperative sera of these patients. In a double-blind study, analysis of serial serum samples from 26 sarcoma patients, 11 normal volunteers, and eight noncancer patients, demonstrated four patterns of circulating IRF: sustained high levels, increasing levels, decreasing levels, and no detectable IRF. Seven of 14 patients who eventually developed metastatic disease demonstrated sustained high or increasing levels of IRF. Eleven of 12 patients clinically free of tumor for five years or more, ten of 11 normal volunteers, and eight of eight noncancer patients had either a pattern of decreasing levels or no detectable IRF. Circulating IRF levels are correlated with the presence of tumor and may be useful in monitoring the clinical course of cancer patients.

Adolescent↗
PubMed1983

Syngeneic humoral immune responses to tumor-associated antigens expressed by K-1735 UV-induced melanoma and its metastases.

An enzyme-linked immunoassay (ELISA) was developed to study syngeneic humoral immune response to a primary tumor and its metastases in the K-1735 ultraviolet light (UV)-induced C3H murine melanoma system. Binding of sera from syngeneic animals previously immunized with primary tumor or metastatic tumor tissue (M-3, M-4) to corresponding 3 M KCl extracts of tumor was significantly greater than binding of control C3H mouse serum. Antibody binding was not significantly reduced by competitive binding with syngeneic murine muscle or liver extracts, indicating the presence of tumor antigen(s) not shared by normal murine tissue. Antibodies to the tumor-associated antigens were selectively removed by competitive binding with syngeneic K-1735 tumor extracts but not by the unrelated 102 murine sarcoma from C57BL/6. However, EL-4 extracts (C57BL/6) did inhibit antibody binding to the primary and both metastases. Further competitive binding studies demonstrated the presence of a common antigen(s) present on the primary tumor and both metastases. We conclude that the K-1735 UV-induced melanoma primary tumor and its metastases express serologically detectable shared antigenic determinate.

Animals↗
PubMed1983

Tumor stasis factor (TSF): a possible mechanism for the regulation of tumor cell proliferation.

A new class of factors that regulates tumor cell division in vitro can be isolated from fresh and cultured tumor cells by 3 M KCl extraction. Tumor stasis factors (TSF) inhibiting cultured tumor cell proliferation were extracted from 8 of 11 fresh human tumors and 2 cultured tumor cell lines. TSF inhibited [3H]Tdr incorporation by allogeneic and autologous cultured tumor cells in a dose-dependent manner. Extracts of normal human tissues and benign tumors did not demonstrate inhibition with the exception of liver. The mechanism of inhibition was cytostatic and not cytotoxic as demonstrated with trypan blue exclusion by tumor cells following TSF treatment, maintenance of intact tumor cell monolayers following addition of TSF, and lack of inhibition of Con A-mediated lymphocyte proliferation by TSF. TSF activity could be reversed by washing for up to 48 hr of incubation and was resistant to heat, pH alterations, reducing agents, proteases, and glycosidases. However, the active moiety bound to lentil lectin and could be purified 80-fold by preparative isoelectric focusing. These factors may represent a novel regulatory mechanism for tumor cell proliferation.

Cell Division↗
PubMed1983

Optical magnification and Doppler ultrasound probe for varicocelectomy.

Varicocelectomy is usually considered a simple urologic operation. However, it is most frequently difficult to visualize and identify the 0.5 mm internal spermatic artery and tiny lymphatic channels when cutting and ligating the internal spermatic veins. The internal spermatic artery and tiny lymphatic channels can be damaged easily, cut, or ligated during this procedure. This occurs surprisingly more often than is realized by the surgeon. By introducing optical magnification or the operating microscope at the time the veins are identified, the surgeon can easily find and dissect off adherent lymphatics or the internal spermatic artery with some microsurgical instruments, and thereby preserve these vessels rather than cut or damage them inadvertently. The sterile Doppler probe also can be utilized to help identify and confirm the location of the internal spermatic artery if it is not easily found in the spermatic cord.

Arteries↗