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Biomedical subjects

D Sun

Publications and source records attributed to D Sun.

At least 379 records · Page 21Linked to original sources
PubMed1992

Prediction of recurrence in giant cell bone tumors by DNA cytometry.

The most interesting therapeutic aspect of giant cell bone tumors is which patients can be cured without a risk of recurrence by intralesional surgery (curettage). To find out the suitability of some DNA cytometric and morphometric parameters for showing differences between this group of patients (n = 9) and those with recurrence (n = 12), the parameters mean ploidy, 2cDI (mean square deviation of the tumor cell DNA content from the normal 2c value), mean nuclear area and its variability were calculated from cytologic specimens prepared by a cell separation technique from formalin-fixed, paraffin-embedded tissues, measuring the values of 100 stromal cells per case by a TV image analysis system. Further measurements were performed on 19 cases of different diseases of the bone and on an additional 17 cases of giant cell tumors without follow-up. The 2cDI allowed us to distinguish the two groups of patients, with and without recurrence, without overlap; even the lowest value for patients with recurrence was higher than the highest value for cured ones. Mean ploidy analysis resulted in a less convincing discrimination of the patients. Mean nuclear area and its variability failed to predict recurrence. Single-cell DNA cytometry provided a parameter, 2cDI, that was able to predict recurrence in patients with giant cell bone tumors with high sensitivity.

Adult↗
PubMed1992

Inhibition of T4 DNA ligase activity by (+)-CC-1065: demonstration of the importance of the stiffening and winding effects of (+)-CC-1065 on DNA.

Non-denaturing gel electrophoresis analysis demonstrates that the stiffening and winding effects of (+)-CC-1065 produce unusual proximal and distal inhibition of T4 DNA ligase-catalysed ligation of covalently modified DNA. (+)-CC-1065 is a potent antitumor antibiotic produced by Streptomyces zelensis. This drug selectively bonds through N3 of adenine in DNA and lies in the minor groove of DNA, reacting in a highly sequence-selective manner. Previous studies (Lee et al., 1991) have shown that (+)-CC-1065 produces bending and winding of DNA. The DNA bending and sequence specificity is mediated by the alkylating 'A' subunit of (+)-CC-1065, while the close van der Waals contacts between the non-alkylating B and C subunits of (+)-CC-1065 and the floor of the minor groove of DNA are responsible for the winding of DNA. Covalent modification of oligomers with (+)-CC-1065 and structurally related drugs leads to preferential inhibition of T4 DNA ligase on the non-covalently modified strand to the 5' side of the covalent adduct site, but enhanced ligation of the covalently modified strand. We speculate that the differential effect on proximal strand ligation is due to a drug-induced winding and helix-stabilizing effect which occurs predominantly to the 5' side of the adduct. In addition to the proximal inhibition of ligation, we also describe a distal inhibition of T4 DNA ligase activity which occurs exclusively with drug-modified oligomers and that, if successful, would result in 180 degrees out-of-phase bent DNA following ligation. In this case, two 25 mers or a 21 plus a 29 mer are inhibited from ligation when modified with (+)-CC-1065. This distal ligation is unique to (+)-CC-1065 and its analogs that cause stiffening of the DNA helix. The (+)-CC-1065-induced stiffening effect was demonstrated using a circularization assay and was found to be associated with the close van der Waals contacts between the inside edge of (+)-CC-1065 and the floor of the minor groove, and also to the benzofuran moiety of (+)-ABC" (Adozelesin), a (+)-CC-1065 analog. We conclude from these studies that the DNA-winding and helix-stabilizing effects of these drug molecules can dramatically affect the efficiency of proximal ligation mediated by T4 DNA ligase, and the unusual helix-stiffening effect of (+)-CC-1065, (+)-AB'C' and (+)-ABC" can stabilize the structure of bent DNA formed by drug modification, which results in distal ligase inhibition.

Antibiotics, Antineoplastic↗
PubMed1991

Analysis of T cell receptor beta chains in Lewis rats with experimental allergic encephalomyelitis: conserved complementarity determining region 3.

This study explores the usage of T cell antigen receptor (TCR) beta chain elements in Lewis rats with experimentally induced allergic encephalomyelitis (EAE). TCRs from 15 different T cell clones and hybridomas derived from animals immunized with myelin basic protein (MBP), and all having specificity for the 21-mer encephalitogenic fragment MBP 68-88, utilized V beta 8.2. In addition, there was a marked conservation of the first two amino acid residues of the junctional complementarity determining region 3 (CDR3) associated with the V beta 8.2 receptors. 12 of 15 contained an aspartic acid followed by serine regardless of the associated J beta element. At the nucleotide level, this conservation of AspSer residues was accomplished with few or no nongermline-encoded nucleotide (N) additions. A similar pattern of AspSer usage and N region nucleotide additions was observed in a number of V beta 8.2 isolates derived from MBP-immunized lymph nodes. In contrast, V beta 8.2 polymerase chain reaction amplified isolates from Lewis T cells activated with concanavalin A or from lymph nodes of complete Freund's adjuvant-immunized animals showed no AspSer utilization (0/31) in the CDR3, and four to nine N region nucleotide additions. We conclude from this finding that AspSer residues in the CDR3, limited N region nucleotide additions, along with V beta 8.2 sequences, contribute to TCR specificity for MBP 68-88. This raises the possibility that encephalitogenic, disease-causing T cells either represent a population that derives from late fetal life or alternatively, that they are rare cells with this particular TCR phenotype contributed to the T cell pool throughout adulthood and are selected by antigen. In either case, the CDR3 AspSer sequences as well as V beta 8.2 sequences are candidates for the receptor target structures recognized by regulator T cells in recovery from and resistance to active EAE. In this respect, a preliminary analysis of TCR utilization in three T cell clones specific for MBP 68-88 isolated from animals recovered from active EAE indicates that while all three use V beta 8.2, only one contains AspSer in the CDR3.

Amino Acid Sequence↗
PubMed1991

CD4-CD8- splenic T cells from Lewis rats recovered from experimental autoimmune encephalomyelitis respond to encephalitogenic T cells that mediate this disorder.

We previously demonstrated that encephalitogenic CD4+ T lymphocytes from the long-term cultured line S1, specific for myelin basic protein, induce a CD8+ T cell population in vivo that protects naive Lewis rats against experimental autoimmune encephalomyelitis caused by S1 cells. In order to determine the contribution of individual T cell population in the development of induced resistance, we have analyzed the in vitro proliferative capacity of phenotypically distinct T cell populations isolated from S1-immunized rats. We found that both CD8+ and CD8-CD4- T cells show striking proliferative responses when stimulated with S1 cells, whereas CD4+ T cells show only minimal responses. In addition, a significant proportion of the CD8-CD4- cells, after stimulated by S1 cells, became CD8+ and had a strong cytolytic activity toward S1 cells. These results suggest a contribution of double-negative splenic T cells in the regulatory circuit associated with autoimmune encephalomyelitis.

Animals↗
PubMed1991

A subset of mouse splenic macrophages can constitutively present alloantigen directly to CD8+ T cells.

About a third of mouse splenic macrophage (M phi) progenitors give rise to cloned progeny that constitutively induce the selective proliferation of naive allogeneic CD8+ T cells in a CD4+ helper cell-independent manner--a response that is inhibited by mAb to the MHC class I molecules present on the M phi. Colony-mixing experiments indicated that the failure of most M phi clones to present allo-Ag was not due to their suppression of the ability of CD8+ cells to respond, nor did the nonpresenting clones interfere with the activity of the allo-Ag presenting M phi. The allo-Ag presenting phenotypes were found to be a stable characteristic in a panel of cell lines derived from individual clones of M phi. Analysis of the cell lines revealed that the differential expression of allo-APC activity could not be attributed to the levels of MHC class I molecules; rather, the cell lines and the primary M phi clones differ in their expression of a cell-associated costimulator molecule that likely functions to induce the expression of the IL-2R on and the secretion of IL-2 from the T cells.

Animals↗
PubMed1991

Constitutive antigen presentation by mouse splenic macrophages is restricted to the progeny of a distinct progenitor population.

Mouse splenic macrophage progenitors differ in their ability to give rise to cloned progeny that constitutively present complex protein antigens to T-cell hybridomas. To determine if the constitutive presentation of diverse antigens is restricted to cells derived from the same subpopulation of progenitors, we expanded macrophage clones into multiple subcultures and compared them for the ability to present different antigens to their respective antigen-specific T-cell hybridomas. Only subcultures derived from the same minority fraction of splenic macrophage progenitors were capable of constitutively presenting the antigens, and the activity of these subcultures was unaffected by the addition of recombinant murine IFN-gamma. This suggests that a specialized sub-population of constitutive antigen-presenting macrophages exists in the spleens of mice.

Animals↗
PubMed1991

The causes and complications of late diagnosis of foreign body aspiration in children. Report of 210 cases.

Late diagnoses of foreign body aspiration were defined as occurring beyond 3 days between the aspiration of the foreign body, or onset of symptoms, and correct diagnosis. We reviewed a total of 210 children with late diagnosis of foreign body aspiration. The causes creating late diagnosis of foreign body aspiration in children were as follows: (1) parental negligence (106/210, 50%); (2) misdiagnosis by the fellow professionals and pediatricians (39/210, 19%); (3) the normal chest roentgenographic findings (29/210, 14%); (4) lack of typical symptoms and signs (26/210, 12%); (5) mismanagement (8/210, 4%); and (6) a negative bronchoscopic finding (2/210, 1%). The most common complications encountered were obstructive emphysema (77/186, 41%), mediastinal shift (63/186, 34%), pneumonia (43/186, 24%), and atelectasis (33/186, 18%) in 186 patients who underwent a chest roentgenographic examination. The incidence of major complications was 64% (48/75) in the children who were diagnosed within 4 to 7 days; however, the complication rate was 70% (39/56) in those with a delay in diagnosis of 15 to 30 days, and 95% (20/21) in the cases with a delay in diagnosis of over 30 days after aspirating the foreign bodies.

Child, Preschool↗
PubMed1991

Enhanced interferon-gamma-induced Ia-antigen expression by glial cells after previous exposure to this cytokine.

Encephalitogenic T cells appear capable of destroying class II major histocompatibility complex (MHC) antigen (Ia)-positive glial cells in the brain, thus accounting for the pathologic activity of these lymphocytes in experimental autoimmune encephalomyelitis (EAE). However, glial cells do not generally express Ia molecules, suggesting that regulation of Ia expression figures prominently in autoimmune diseases of the central nervous system. In studies to understand the regulatory mechanisms involved in Ia expression, a glial cell clone generated from the brains of neonatal Lewis rats (F10 clone) readily expressed class II major histocompatibility (Ia) antigens after stimulation by interferon-gamma (IFN-gamma) at doses from 10 to 100 units/ml. Level of the antigen decreased gradually within 5-7 days after cultures were depleted of the cytokine. Reexposure of the cells to the IFN-gamma at 100-fold lower doses induced a stronger Ia response than did the initial exposure. F10 cells also expressed Ia when they were cultured with small numbers of syngeneic T lymphocytes, either proliferating or nonproliferating. Proliferating T cells had direct Ia-inducing activity, whereas nonproliferating T cells had this effect only when they were added to cultures with small amounts of T cell-specific antigen. Moreover, Ia-inducing effects of IFN-gamma on F10 cells were also greatly enhanced when these cells were preexposured to T cells. Our results suggest that initial exposure to IFN-gamma or T cells enhances the Ia responsiveness of glial cells to further stimulation with the cytokine.

Animals↗
PubMed1991

Characterization of monoclonal antibodies against alpha-hemolysin of Escherichia coli.

Monoclonal antibodies (MAbs) were raised against native and denatured alpha-hemolysin (HlyA) of Escherichia coli. Binding of the MAbs to native, denatured, and erythrocyte-complexed active wild-type hemolysin and mutant derivatives was tested. All 15 MAbs analyzed bound to native hemolysin, even when the toxin was complexed with human erythrocytes. While some MAbs were unable to bind to a specific native mutant hemolysin, others could not even bind to mutant hemolysin carrying deletions remote from their actual binding sites. A rough determination of the binding sites of 15 MAbs on HlyA was performed by Western immunoblot analysis using CNBr fragments of HlyA and mutant hemolysin proteins. Interestingly, the binding sites of the MAbs against native hemolysin seem to be more randomly distributed on HlyA than are those of MAbs against denatured hemolysin. Three MAbs inhibited the hemolytic activity significantly. Two of these MAbs bound to the hydrophobic region, and the other one bound to the repeat domain of HlyA. The use of synthetic peptides from these regions allowed determination of the linear epitopes for two of these MAbs.

Animals↗
PubMed1991

Analysis of transcriptional control of the gerD spore germination gene of Bacillus subtilis 168.

The gerD locus of Bacillus subtilis comprises a single gene whose function is essential for the germination of B. subtilis spores in media containing asparagine, glucose, and fructose. The expression of gerD has been characterized by using a chromosomal lacZ fusion to the gerD promoter. The promoter is switched on at the same time as the synthesis of glucose dehydrogenase, 2.5 h after sporulation has been initiated in the developing forespore. The gerD gene is not expressed in spoIIB or spoIIIA, -IIIB, -EIII, -FIII, or -IIIG mutants, but it is expressed in spoIIIC and -IIID and spoIVA mutant backgrounds. The in vivo transcriptional start point of the gene has been mapped by primer extension analysis, and sequences upstream from the start point show considerable homology with the promoter consensus sequences recognized by RNA polymerase containing the forespore-specific sigma factor sigma G (E sigma G). gerD is transcribed in vitro by E sigma G with a similar if not identical start point to that found in vivo, and expression of the gene can be rapidly induced in vegetative cells following the induction of sigma G synthesis. These results indicate that gerD is another member of the sigma G regulon, which includes a number of genes expressed only in the forespore compartment of sporulating cells of B. subtilis.

Bacillus subtilis↗
PubMed1991

Effect of chromosome location of Bacillus subtilis forespore genes on their spo gene dependence and transcription by E sigma F: identification of features of good E sigma F-dependent promoters.

Translational lacZ fusions to forespore genes of Bacillus subtilis were not expressed in spoIIAC (sigma F) or spoIIIE mutants when the lacZ fusions were integrated at the loci of the same genes or at the SP beta locus. However, some of these genes, including gerA, gpr, spoIIIG (sigma G), and sspE, were expressed in spoIIIE mutants and spoIIIE spoIIIG double mutants (but not in spoIIAC mutants) when the lacZ fusions were integrated at the amyE locus. When tested, the beta-galactosidase made in these mutants was found only in the forespore, and the 5' ends of the mRNAs produced in these mutants were identical to those in a Spo+ background. Analysis of the in vitro transcription of forespore genes by RNA polymerase containing sigma F (E sigma F) revealed a direct correlation between good in vitro transcription by E sigma F and expression at the amyE locus in spoIIIE mutants. This result suggests that forespore genes are transcribed by E sigma F in spoIIIE and spoIIIE spoIIIG mutants. Comparison of the promoter regions of genes transcribed well and poorly by E sigma F in vivo and in vitro showed that good transcription by E sigma F was correlated with G residues at positions -15 and -16, a purine residue at position -13, and a T residue at position -7 relative to the start site of transcription. The importance of these residues in sigma F recognition was confirmed by analysis of the E sigma F-dependent transcription in vivo and in vitro of mutant ssp genes.

Bacillus subtilis↗
PubMed1991

Inhalation of foreign bodies in Chinese children: a review of 400 cases.

A retrospective review of 400 Chinese children who had inhaled foreign bodies was undertaken. There has been a yearly increase in the total number of cases of airway foreign bodies removed in our hospital. Fifty-eight percent of the children presenting were from the countryside; 42% were townspeople. Approximately 90% of the patients were under 3 years of age, with the peak incidence of foreign body inhalation occurring between 1 and 2 years of age (57.8%). The male-female ratio was about 1.2:1. About 95% of the removed foreign bodies were organic in origin. The majority of the foreign bodies were found most often in the right bronchial tree (46%). A positive history of foreign body inhalation was obtained in 98% of the cases. Twenty-eight percent of the children presented at the hospital within 24 hours, 71% within 1 week, and 29% more than 1 week after inhaling the foreign body. The most common presenting symptoms of laryngotracheal foreign bodies were cough, wheezing, dyspnea, and hoarseness; those of bronchial foreign bodies were cough, wheezing, decreased air entry, and rhonchi. More than two-thirds of the children with larygotracheal foreign bodies had normal x-ray findings. The most common fluoroscopic findings in those children with bronchial foreign bodies were mediastinal shift (36.8%), obstructive emphysema (35.7%), and normal findings (35%). A total of 348 (87%) bronchial foreign bodies were removed by rigid bronchoscopy (81%), rod-lens bronchoscopy (5%), and spontaneous expulsion (1%); 52 (13%) laryngeal and tracheal foreign bodies were removed by direct laryngoscopy (12%) and tracheotomy (1%). A single endoscopic procedure successfully removed 92.5% of 400 foreign bodies detected in the airways. One child died during bronchoscopy, for a mortality rate of 0.25%.

Adolescent↗
PubMed1990

Establishment of a permanent rat brain-derived glial cell line as a source of purified oligodendrocyte-type 2 astrocyte lineage cell populations.

A permanent glial cell line (L3) has been established from mixed glial cultures obtained from neonatal rat forebrain by repetitive passaging and selection of the process-bearing cells growing on top of a flat cell monolayer. Continuous propagation of the process-bearing cells was supported by the flat cells, of presumed astroglial origin, which were present in negligible amounts following each passage but then grew and formed a basal, feeder layer. Throughout a culture period of over 2 years, the L3 cells have maintained a stable morphological and antigenic phenotype. In serum-containing culture medium, most of the process-bearing cells expressed at the same time features of immature oligodendrocytes (O4 positivity) and of astrocytes [glial fibrillary acidic protein (GFAP) positivity]. A smaller proportion of them was labeled by the monoclonal antibody LB1. LB1+ or O4+ cells were rarely GFAP-, and GFAP+ cells were rarely LB1- or O4-. GalC+ oligodendrocytes were seen only occasionally, but the proportion of these cells increased up to 30% upon culturing in chemically defined medium containing 0.5% fetal calf serum. The L3 process-bearing cells accumulated the neurotransmitter gamma-aminobutyric acid (GABA), expressed the proteoglycan chondroitin sulfate, and responded to the mitogenic action of platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF). All these properties are characteristic of cells belonging to the O-2A (oligodendrocyte-type 2 astrocyte) cell lineage. The L3 flat cells were largely negative for the glial markers tested, but resembled type 1 astrocytes in their ability to support the growth of O-2A lineage cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Variation of secretory antibodies in parotid saliva to human immunodeficiency virus type 1 with HIV-1 disease stage.

The secretory immune response to pathogens of the gut-associated lymphoid tissue is often independent of the systemic response. We investigated and compared the presence of antibodies to human immunodeficiency virus type 1 (HIV-1) antigens in parotid saliva and serum by Western blotting in 22 HIV-1-infected individuals. Antibodies to the HIV-1 envelope antigen gp160 were detected in saliva samples from 21 of 22 individuals and in serum from all individuals who were classified as CDC Group II, III, or IV. Antibody titers to gp160 were approximately 3000 times higher in serum than in saliva. Antibodies to viral core antigen p24 were detected in 6 of 7 Group II individuals in saliva and in 7 of 7 in serum. Antibodies to p24 were not found in the parotid saliva, but were detected in the sera of 3 of 3 Group III and 11 of 12 Group IV patients. The absence of secretory antibodies to HIV-1 core antigen p24 was correlated with CD4+ cell counts of less than 200/mm3. The results suggest that loss of secretory anti-p24 antibodies may be an early sign of progression to higher CDC clinical stages in HIV-1-infected individuals.

Blotting, Western↗
PubMed1990

The regulation of transcription of the gerA spore germination operon of Bacillus subtilis.

The gerA operon of Bacillus subtilis 168 comprises three genes concerned with the triggering of spore germination by L-alanine and its analogues. The expression of this operon has been characterized using chromosomal lacZ fusions to the gerA promoter. The gerA promoter is switched on 2.5-3 hours after the initiation of sporulation, in parallel with glucose dehydrogenase. A high proportion of the gerA-driven beta-galactosidase detected in sporulating cells is found in the mature spore; the gerA promoter is therefore active in the forespore compartment of the sporulating cell. The gerA promoter is not expressed in spoO, spoII or spoIIIA, B, E and G mutant backgrounds, but is expressed in spoIIIC and D and in spoIV and V mutants. The in vivo transcriptional startpoint of the operon has been mapped by primer extension experiments; sequences upstream from this startpoint show significant homology with recognition sequences for RNA polymerase containing sigma G (E sigma G). The gerA operon was transcribed in vitro by E sigma G with a startpoint identical to that used in vivo, and expression of the gerA operon was rapidly induced in vegetative cells by induction of sigma G synthesis. These data indicate that the gerA operon is an additional member of the sigma G regulon, which includes a number of genes expressed in parallel only in the forespore compartment of sporulating B. subtilis cells.

Bacillus subtilis↗
PubMed1990

Salivary antibodies to human immunodeficiency virus type 1 in a phase I AIDS vaccine trial.

HIV-1-specific secretory antibodies may be a desirable outcome in individuals receiving AIDS vaccines. We investigated parotid and whole saliva samples for HIV-specific antibodies collected from five volunteers who received a recombinant HIV-1 envelope glycoprotein (rgp160) vaccine. Ten healthy, adult volunteers received intramuscularly either three doses of rgp160 (40 or 80 micrograms), a hepatitis B vaccine, or a placebo on days 0, 30, and 180. Saliva samples were collected on days 0, 28, 60, 120, 194, and 270 from the volunteers. All volunteers were negative for serum HIV antibodies by ELISA (Abbott). By Western blotting, serum antibodies to envelope antigens were demonstrated in one of three individuals who received the low dose vaccine and two of two who received the high dose. Antibodies to gp160 were detected in whole saliva on day 194 from one of these individuals by Western blotting. Parotid saliva collected on all dates did not contain detectable HIV-specific antibodies. The finding of HIV-1-specific antibodies in whole saliva following vaccination may indicate that development of mucosal immunity is possible.

Blotting, Western↗
PubMed1990

[The effect of the emergent vaccination using DTP vaccine to control the outbreak of pertussis].

This paper reports the results of the effects of the emergent vaccination with DTP vaccine to control the outbreak of pertussis. Objects of observation were 4-6 years old children of kindergarten having pertussis vaccination and exposed history. The children were at random divided into two groups: 76 children were vaccinated with DTP vaccine and 72 children were inoculated with placebo. The results of observation: there was no strong and abnormal reaction in the vaccinated children; two weeks and three months after emergent vaccination with DTP vaccine, GMT antibody of pertussis were increased 18.14 and 4.73 times more than before vaccination respectively. There were no differences of the attack rates average days from vaccination to onset, average days of cough (P greater than 0.2) within 21 days after vaccination between the two groups. The protection rate was 62.11% three months after vaccination. The results showed that the emergent vaccination with DTP to control the outbreak of pertussis was safe and effective.

Child↗