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Biomedical subjects

J Gan

Publications and source records attributed to J Gan.

At least 91 records · Page 5Linked to original sources
PubMed1994

Melatonin receptor-mediated inhibition of cyclic AMP accumulation in chick retinal cell cultures.

Melatonin receptors were characterized in cultured neurons and photoreceptors prepared from chick embryo retina. Cultured cells contained high-affinity 2-[125I]iodomelatonin binding sites (KD = 41.6 pM), similar to those in intact retina. The effects of melatonin and related indoles on cyclic AMP accumulation were examined. Melatonin (10(-7) M) had no effect on basal or K(+)-stimulated cyclic AMP accumulation, but inhibited forskolin-stimulated cyclic AMP accumulation by approximately 50%. Melatonin inhibited forskolin-stimulated cyclic AMP accumulation in the presence or absence of the cyclic nucleotide phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, suggesting an effect on cyclic AMP synthesis rather than degradation. Half-maximal inhibition was observed at 5.9 x 10(-10) M melatonin. The relative order of potency among melatonin analogues was 2-iodomelatonin > melatonin approximately 6-chloromelatonin > or = 6-hydroxymelatonin > N-acetylserotonin approximately 5-methoxytryptophol > serotonin. The EC50 value for inhibition of cyclic AMP accumulation by 2-iodomelatonin (36.7 pM) was comparable to the KD value for binding of the radioligand, suggesting that the binding sites represent functional receptors. The inhibitory effect of melatonin was antagonized by the putative melatonin antagonists luzindole, N-acetyltryptamine, and N-(2,4-dinitrophenyl)-5-methoxytryptamine, with estimated KB values of 0.12, 0.17, and 1 microM, respectively. At a concentration of 10 microM, N-(2,4-dinitrophenyl)-5-methoxytryptamine significantly inhibited forskolin-stimulated cyclic AMP accumulation when added alone; at 30 microM, luzindole and N-acetyltryptamine also had significant inhibitory effects. The inhibitory effect of melatonin was blocked by pretreatment with pertussis toxin.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Methyl-3-isobutylxanthine↗
PubMed1994

Treatment of neuroblastoma patients with antiganglioside GD2 antibody plus interleukin-2 induces antibody-dependent cellular cytotoxicity against neuroblastoma detected in vitro.

Therapy of neuroblastoma patients with interleukin (IL)-2 activates effector cells capable of lysing tumor cells in vitro. When tumor cells are pretreated with certain monoclonal antibodies (MoAb), these in vivo activated effectors show augmented tumor lysis via antibody-dependent cellular cytotoxicity (ADCC). This study presents immunological analyses of serial blood samples from two refractory neuroblastoma patients who received combined in vivo therapy with murine anti-ganglioside GD2 monoclonal antibody 14.G2a and IL-2. These studies were designed to determine whether conditions that induce ADCC in vitro can be generated in vivo by combined therapy with IL-2 and MoAb. As shown previously, administration of IL-2 dramatically augments the ability of peripheral blood mononuclear cells (PBMC) to mediate ADCC. In addition, we demonstrate here that sera, obtained 1 h after infusion of 14.G2a, provides an effective source of functional antibody for ADCC mediated by PBMC from healthy donors. Finally, effective ADCC-mediated killing of neuroblastoma target cells was also achieved in vitro following IL-2 plus 14.G2a treatment when patients' effector cells were combined with patients' serum, as the source of 14.G2a antibody. These results indicate that this combination of IL-2 and 14.G2a generates conditions within the peripheral blood of pediatric neuroblastoma patients that enable their own lymphocytes to mediate antibody-dependent cellular cytotoxicity sufficient to effectively kill neuroblastoma cells in vitro.

Antibodies, Monoclonal↗
PubMed1994

Potential to involve multiple effector cells with human recombinant interleukin-2 and antiganglioside monoclonal antibodies in a canine malignant melanoma immunotherapy model.

Human tumors originating from neuroectodermal cells such as malignant melanoma and neuroblastoma express high levels of disialogangliosides GD2 and GD3, making these antigens ideal for targeting by monoclonal antibodies (Mabs). The purpose of this study was to investigate expression and targeting of gangliosides on canine melanoma. Using immunohistochemical methods, we analyzed the expression of disialogangliosides GD2 and GD3 on canine oral malignant melanomas with murine Mabs 14.G2a and R24 that recognize GD2 and GD3 disialogangliosides, respectively, on human tumors. We also assessed the ability of Mab 14.G2a (and its mouse-human chimera, ch 14.18) to mediate antibody-dependent cellular cytotoxicity (ADCC) in vitro against a canine malignant melanoma cell line with human recombinant interleukin-2 (IL-2) activated canine peripheral blood lymphocytes (PBL), or canine neutrophil effector cells. Our data show that Mabs 14.G2a and R24 recognized fresh frozen canine oral melanoma. Mabs 14.G2a or ch 14.18, or IL-2, potentiated lysis of the canine malignant melanoma cell line by canine PBL. The killing effect observed using the combination of either Mab with IL-2 was additive. Mab 14.G2a mediated potent ADCC of canine melanoma by canine neutrophils. These studies indicate that disialogangliosides are expressed on fresh canine melanoma cells. Mabs reactive with these antigens can target and trigger tumor killing by multiple canine effector populations and IL-2 can potentiate these effects by canine lymphocytes. Thus, canine oral malignant melanoma, a spontaneously occurring, metastatic cancer in the dog, may be a relevant animal model to investigate combination immunotherapy using antitumor Mab and IL-2.

Animals↗
PubMed1993

In vitro inactivation of glucose-6-phosphate dehydrogenase from human red blood cells by acrolein: a possible biomarker of exposure.

We have investigated the possibility of utilizing glucose-6-phosphate dehydrogenase (G6PD) as a macromolecular (biological) marker of acrolein exposure. The result showed a dose-dependent inactivation of the erythrocyte G6PD in situ or as a purified enzyme from human erythrocytes or yeast. Amino acid analysis on the chemically modified yeast G6PD showed a formation of a lysine adduct which is probably linked to the inactivation.

Acrolein↗
PubMed1992

Translational options for the pir gene of plasmid R6K: multiple forms of the replication initiator protein pi.

The autogenously controlled pir gene of plasmid R6K was believed to encode a single polypeptide that plays multiple roles in the plasmid's biology. We have isolated an opal (op) mutant at the 18th codon of the pir coding frame which does not totally abolish translation of pir mRNA. In extracts of cells containing this mutation two translational products (35 kDa and 30.2 kDa) have been detected. We propose that the 35-kDa polypeptide produced by the pir18 op mutation contains Trp substituted for Arg18 as the result of an opal readthrough. Translation, which results in the 30.2-kDa polypeptide, originates downstream from the UGA stop signal created by the mutation. Moreover, we realize now that the 30.2-kDa polypeptide is also produced in cells containing a wild-type (wt) pir gene. The shorter variant of the pi protein lacks replication initiation and inhibition functions, as well as autorepressor activity in vivo. We also show that an in-frame fusion of seven N-terminal codons of the trpE gene with a pir gene lacking the first two codons produces two polypeptides which replace the 35-kDa pi protein and are of similar molecular weight. Thus, at least three options exist in the translation of the wt pir mRNA. Start codons are most likely at codon positions 1, 6 or 7, and 36 or 38. Each of these five AUG codons is preceded by a consensus ribosome-binding site (RBS).

Bacterial Proteins↗
PubMed1991

Relationship between fasting plasma insulin and ambulatory blood pressure in young, non-obese normotensive subjects.

In order to investigate the relationship between ambulatory blood pressure values and fasting plasma insulin, non-invasive, 24-h blood pressure monitoring was performed in 32 young normotensive males. Systolic and diastolic blood pressures were averaged for awake and asleep periods. Fasting plasma insulin levels correlated significantly with both asleep (r = 0.61; P less than 0.001) and awake (r = 0.44; P less than 0.02) systolic blood pressure, but not with casual systolic blood pressure (r = 0.27). There were no significant associations between awake, asleep and casual diastolic blood pressure, and fasting plasma insulin levels (r = 0.15, 0.05 and 0.21, respectively). These results support the hypothesis that insulin may be a physiological determinant of blood pressure.

Adult↗
PubMed1991

K(+)-evoked depolarization stimulates cyclic AMP accumulation in photoreceptor-enriched retinal cell cultures: role of calcium influx through dihydropyridine-sensitive calcium channels.

The effect of membrane depolarization on cyclic AMP synthesis was studied in glia-free, low-density, monolayer cultures of chick retinal photoreceptors and neurons. In photoreceptor-enriched cultures prepared from embryonic day 6 retinas and cultured for 6 days, elevated K+ concentrations increased the intracellular concentration of cyclic AMP and stimulated the conversion of [3H]adenine to [3H]cyclic AMP. The K(+)-evoked increase of cyclic AMP accumulation was blocked by omitting CaCl2 from the incubation medium, indicating a requirement for extracellular Ca2+. Stimulation of cyclic AMP accumulation was also inhibited by nifedipine, methoxyverapamil, Cd2+, Co2+, and Mg2+, and was enhanced by the dihydropyridine Ca2+ channel agonist Bay K 8644. The enhancement of K(+)-evoked cyclic AMP accumulation by Bay K 8644 was antagonized by nifedipine. Thus, Ca2+ influx through dihydropyridine-sensitive channel is required for depolarization-evoked stimulation of cyclic AMP accumulation in photoreceptor-enriched cultures.

1-Methyl-3-isobutylxanthine↗
PubMed1989

A unifying parameter to describe the clinical mechanics of hydrogel contact lenses.

We studied the relation of postlens squeeze pressure to the clinical fit of hydrogel contact lenses. This relation was statistically significant (p less than 0.01) for a variety of contact lenses currently prescribed over a range of thickness, water content, back central optic radius, diameter, and back vertex power. The squeeze pressure provided a useful parameter to describe and compare the clinical fit of hydrogel contact lenses and a model with which the fit of a lens to an eye could be predicted. There was virtually no movement of the contact lenses across the eye when the squeeze pressure was more than -14 mm H2O (-1370 dynes/cm2). However, the mechanics of the lens motion when the squeeze pressure was less than this critical pressure appeared to be related to a more complicated combination of the squeeze pressure and the other forces in the eye-lens system.

Contact Lenses, Hydrophilic↗
PubMed1987

Application of monoclonal antibodies for affinity electrophoresis.

Monoclonal antibodies (MoAbs) to human alpha-fetoprotein (AFP) produced by 13 different murine hybridoma clones were used in affinity electrophoresis. The binding of MoAbs to AFP was studied in different experimental systems: crossed affinity immunoelectrophoresis, rocket affinity immunoelectrophoresis and zone affinity electrophoresis in agarose. Experimental data indicated that MoAb-AFP and AFP-MoAb-AFP complexes were formed during electrophoresis. Affinity electrophoresis seems to be a convenient and sensitive method for studies on the reaction of monoclonal antibodies with their antigens. In addition, the technique offers the possibility of calculating the dissociation constants of MoAb-AFP and of AFP-MoAb-AFP complexes and evaluating the reaction of MoAbs with different antigenic determinants.

Antibodies, Monoclonal↗
PubMed1987

The influence of thymus factor X (TFX) and of anti-TFX globulin on the growth of Lewis lung carcinoma in mice.

The influence of thymus factor X--TFX (Polfa) and an anti-TFX rabbit gammaglobulin (RATFX) on the growth of Lewis lung carcinoma in mice was studied. The preparations were administered subcutaneously into the peritumoral region. Tumor growth was significantly retarded in the RATFX-treated groups, while a low dose TFX therapy was ineffective. No significant differences in peritumoral inflammatory reaction in treated and untreated mice were found.

Animals↗
PubMed1985

Differential binding of soybean agglutinin to human neuroblastoma cell lines: potential application to autologous bone marrow transplantation.

Normal human bone marrow cells were mixed with neuroblastoma cells from four different human cell lines, and the cell mixtures were separated by differential agglutination with soybean agglutinin (SBA). The unagglutinated cell fraction, previously shown to be highly enriched for the hematopoietic pluripotential stem cells and capable of reconstituting lethally irradiated adult patients with acute leukemia, was further fractionated by affinity chromatography on the lectin conjugated to Sepharose 6MB beads. Two independent assays, one using radiolabeling of the tumor cells and the other based on cloning of the neuroblastoma cells on agar, showed that the agglutination step alone removes 64-76% of the radiolabeled neuroblastoma cells and 85-98% of the clonogenic cells from the tumor/bone marrow cell mixture. Passage of the unagglutinated radiolabeled cells through SBA-Sepharose columns results in further purging of 28-53% of the neuroblastoma cells. Thus a combination of the two methods affords only one-log depletion for the neuroblastoma cells, compared to a three-log depletion achieved for a T-cell leukemia line CEM tested in parallel. It seems therefore that the agglutination technique, or the use of SBA-Sepharose columns, can be used only as a preliminary step for the purging of neuroblastoma cells from involved human bone marrow preparations. Staining with fluorescein isothiocyanate-conjugated SBA of nine different neuroblastoma cell lines, including the four tested in the fractionation studies, showed that more than 98% of the cells, of all the cell lines tested, specifically bind to the lectin, whereas no specific binding can be detected on the stem cell-enriched bone marrow cell fraction. However, the total number of receptors on the neuroblastoma cells is small compared to that of line CEM or normal granulocytes, which are strongly agglutinated by SBA. It seems therefore that the quantitative difference in the total number of SBA receptors is a crucial factor for purging by the agglutination technique or by affinity chromatography. Although these results show limitations to the use of both methods, this study establishes that all neuroblastoma cell lines tested express receptors for the lectin. Improved purging of neuroblastoma cells may possibly be achieved by targeting SBA-bound toxins or magnetic spheres to these receptors.

Agglutination↗