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Biomedical subjects

G G Miller

Publications and source records attributed to G G Miller.

At least 91 records · Page 5Linked to original sources
PubMed1991

[Interactions between Mycoplasma and human and animal viruses. Ultrastructural analysis].

Presented are the data on the ultrastructural analysis of interaction between mycoplasma and certain cancerogenic and infectious viruses in humans and animals. Revealed are spontaneous associations of mycoplasma with viruses of cattle leukemia, T-cell human leukemia and with a representative of Bunyaviruses. Immediate interaction of these agents is found possible. Simulated complexes of mycoplasma with infectious viruses are developed. Electron microscopy on supramolecular levels revealed immediate interaction of different agents in membranes. Some methodological procedures help to reveal that the interaction of M. pneumoniae and A. laidlawii with orthomyxo-, paramyxo and togavirus is of specific character and is realized as receptor ligand form due to the affinity in the receptor requirements of these pathogens. This property as well as a bequeath distribution and frequent association in respiratory infections enable one to suggest the possibility of their immediate interactions in a host body.

Animals↗
PubMed1991

Effect of fibroblast implants on wound healing of irradiated skin: assay of wound strength and quantitative immunohistology of collagen.

The role of dermal fibroblasts in the expression of radiation-induced damage to the skin was studied. Fibroblasts from neonatal mice were cultured, harvested, and injected into full-depth surgical incisions in the dorsal area of mouse skin, which had been previously locally irradiated by 18 Gy X rays. As a control, cells irradiated with a dose of 20 Gy were also injected. The effect of radiation and fibroblast implants on the gain of skin wound strength was assayed. In an additional experiment freshly isolated cells were implanted. Two weeks following wounding the irradiated skin had reached only about a third of the strength of unirradiated skin. A significant increase of wound strength in irradiated skin was observed when 1.5-2 x 10(6) cultured fibroblasts or freshly isolated fibroblasts were injected into the 20-mm-long wound bed. Irradiated cells had significantly less effect. This suggests that implanting isolated syngeneic cells may "rescue" wounds from the effect of prior irradiation. Semiquantitative immunohistology of types I and III collagen was performed in parallel using a video image digitizing system. Levels of both types I and III collagen were altered in the dermis and the wound tissues in irradiated skin, but the implant of cultured fibroblasts did not affect notably the total levels and the disposition of the two collagen isotypes.

Animals↗
PubMed1989

Insulin B chain functions as an effective competitor of antigen presentation via peptide homologies present in the thymus.

The B chain of mammalian insulins contains appropriately spaced amino acids that predict recognition by T cells. However, all T cell clones from an HLA-DR1, Dw6 diabetic donor recognize epitopes associated with the A chain, and the B chain was found to inhibit these responses. Effective intramolecular competition at the level of the APC, not a direct effect on the T cell, is responsible for the inhibition. Insulin B chain contains two clusters of amino acid homology with the TCR beta chain and B chain peptides lacking these clusters do not compete for antigen presentation. A hole in the repertoire for T cells that recognize this portion of the insulin molecule may arise in the thymus by deletion of T cells that recognize similar peptides.

Amino Acid Sequence↗
PubMed1989

Quantitation of hypoxia in multicellular spheroids by video image analysis.

Polyclonal antibodies to a series of radiation chemically produced 2-nitroimidazole-protein adducts have been used in the indirect immunofluorescent detection of hypoxic cells in EMT6/Ed spheroids. The spheroids were incubated with the radioactive 2-nitrioimidazoles under atmospheres of air, several intermediate oxygen concentrations, and nitrogen. The fluorescence intensity of the marker across radii of spheroids labeled at various oxygen tensions, as measured by video image analysis, paralleled the autoradiographic grain density. Thus, we conclude that the immunohistochemical approach provides a quantitative measure of the distribution of 2-nitroimidazole adducts in spheroids. The advantages of speed, technical simplicity, economy, and independence from the requirement of radiolabeled precursor render the fluorescent marker of potential use in the clinical detection of cellular hypoxia and tissue ischemia.

Autoradiography↗
PubMed1989

Immunohistochemical quantitation of three collagen isotypes in perfused areas and nonperfused foci of the lungs of irradiated mice.

Collagen isotypes I, III, and IV were quantitated by video image analysis of fluorescent-antibody-stained lung tissue sections from control and irradiated C57L/J and BALB/c mice. The perfusion status of lungs was determined by injecting colloidal carbon into the hepatic vein immediately prior to sacrificing the animals. Well-perfused parenchymal regions turned black, whereas nonperfused areas remained pale. Previous histological studies indicated substantial differences in the types of lesions found in the lungs of these two strains. C57L/J mice develop extensive and persistent contracted fibrosis. In lung sections of C57L/J mice examined 28 weeks after a dose of 11 Gy X rays, all three collagen isotypes were significantly elevated to levels 37-51% higher than age-matched control values in perfused regions of lung. In nonperfused areas, which had the histological appearance of contracted scar tissue, the three collagen isotype levels were further elevated to values 83-90% greater than controls. This finding suggests that in C57L/J mice, an elevation of each or all of the three collagen isotypes to levels approximately 45% greater than controls is consistent with continued pulmonary function during the intermediate phase of lung damage, whereas areas of parenchyma containing isotype levels in excess of 185% of control values coincide with functionally deficient regions. BALB/cCr//Alt. mice examined 28 weeks subsequent to 14.5 Gy X rays had a variety of visible lesion, most of which were nonperfused. In addition, one-quarter of nonperfused acini had no visible lesion. In perfused areas, the three isotypes were increased to 119-132% of control levels, with a further, significant (P less than 0.05) increase to 128-144% of control values in nonperfused parenchyma. Nonperfused areas were not characterized by contracted fibrosis; however, it would appear that the threshold level for collagen elevation associated with functional compromise during intermediate phase lies in the region of 130%. For BALB/c/J mice, 1 year after 9 Gy X rays, perfused areas of lung contained control levels of the three collagen isotypes, while nonperfused areas had isotype levels 119-131% of control values. Two of seven animals died at 41 weeks, but we were unable to ascertain collagen levels, since the lungs were not infused with colloidal carbon.

Animals↗
PubMed1988

Antigen processing and the human T cell receptor repertoire for insulin.

Three human T cell lines specific for the A loop of beef insulin were studied to determine the requirements for Ag processing. The data show that the conformation of the A loop of insulin is required for recognition and that the B chain of insulin per se is not necessary for this response. Processing of native insulin was required for responses of all three T cell lines; however, each displayed a different pattern of sensitivity to inhibition of processing and aldehyde fixation of APC. A peptide comprised of two disulfide-linked A chains was partially stimulatory when presented by fixed APC whereas A chain monomers and disulfide-linked A and B chain peptides were not. The response to native insulin, peptides, and A chain dimers was sensitive to chloroquine suggesting that none of these moieties is the terminal processed peptide recognized by insulin immune T cells. The unique patterns of fine specificity, processing requirements, and recognition of aldehyde-fixed antigen-MHC for each T cell line suggest the hypothesis that Ag processing leads to heterogeneity of the T cell repertoire for a single epitope of insulin.

Amino Acids↗
PubMed1988

Functional versus phenotypic analysis of T cells in subjects seropositive for the human immunodeficiency virus: a prospective study of in vitro responses to Cryptococcus neoformans.

We performed a prospective study of 50 subjects at high risk for human immunodeficiency virus (HIV) infection to determine if assays of antigen-specific T cell function provide an earlier indication of future progression to AIDS or a better assessment of immune function than do current methods of evaluation. We measured in vitro T cell responses to Cryptococcus neoformans and tetanus toxoid, response to mitogens, HIV p24 antigenemia, and clinical parameters. Progression to AIDS was significantly associated with loss of T cell response to cryptococci (P = .015), HIV antigenemia (P = .001), and low CD4+ cell numbers (P = .001). Most importantly, we found that loss of antigen-specific responses to cryptococci and tetanus can occur before changes in CD4 cell number. Abnormal response to mitogens and marked depletion of CD4+ cells were late signs of progressive HIV infection. Measurement of antigen-specific T cell function may be useful for assessing the efficacy of antiviral therapy in HIV infection before onset of symptoms.

Acquired Immunodeficiency Syndrome↗
PubMed1988

Radiation-induced changes in collagen isotypes I, III, and IV in the lung of LAF1 mouse: effects of time, dose, and WR-2721.

Alterations in the amount and distribution of pulmonary connective tissue are commonly observed subsequent to thoracic radiotherapy. The extent to which these changes are important in the expression of radiation damage and its repair remains unclear. We have quantitated changes in the parenchymal levels of collagen types I, III, and IV in the lungs of LAF1 mice at intervals to 1 year, following doses of 0-14 Gy, 300 kV X rays, or 0-18 Gy in the presence of the radioprotective compound, WR-2721. The method of quantitation, which involves video image analysis of fluorescent antibody stained, cryostat tissue sections, provides both quantitative and morphological information for the three collagen isotypes. Type I collagen peaked in tissue content at 15 and 30 weeks postirradiation (p.i.), with transient return to control values 20-25 weeks p.i. Type III collagen peaked at 15 and 25 weeks p.i. and declined in tissue content at 20 and 30 weeks. Type IV peaked 15-20 weeks following irradiation, returned to control levels at 25 weeks, and reached a plateau above control values after 30 weeks. Fluctuations in collagen levels in the parenchyma were dose dependent but were not simultaneous, indicating a radiation response characterized by alpha-chain-specific regulation of collagen biosynthesis and breakdown. In general, WR-2721, which enhanced postirradiation survival (DMF, 1.3), reduced the magnitude and altered the timing of collagen fluctuations; again, the effects were type specific. The results clearly demonstrate that the postirradiation response of the connective tissue is dose dependent, is specific to each macromolecule, and involves both deposition and removal of extracellular matrix. These processes are independently influenced by the presence during irradiation of WR-2721.

Amifostine↗
PubMed1988

[Isolation and characteristics of the bacteriophage from the vaccine strain Tohama phase I].

Some properties of bacteriophage phi T isolated from the vaccine strain Bordetella pertussis Tohama phase I and propagated in Bordetella parapertussis 504 cells are presented. Phage phi T belongs to the IV group in accordance with Tikhonenko classification. The diameter of head and length of noncontractile tail sheath are 49.5 +/- 0.5 and 145 +/- 7 nm, respectively. Diameter of the tail sheath is 3.2 +/- 0.6 nm. Molecular mass of the phage DNA is 37 +/- 3 kb. Population of phi T phage is polymorphous and consists of particles the genomes or which vary from each other by the "insert" located 6.8 +/- 0.6 kb from the end of molecule. The blot hybridization has demonstrated that the bacteriophage genome is not inserted into the chromosome of the lysogenic strain. Autonomous location of the phage genome in the host cell is suggested. The temperature and hydrogen ions concentration effects on bacteriophage phi T stability were studied. The conditions for phage suspension storage are described.

Bacteriophages↗
PubMed1987

Insulin-specific human T cells. Epitope specificity, major histocompatibility complex restriction, and alloreactivity to a diabetes-associated haplotype.

T cells from an insulin-treated diabetic (ML, HLA DR1, w6) were stimulated in vitro with insulin, cloned at limiting dilution, and examined for their fine specificity and genetic restriction. T cell lines (TCL) derived from beef insulin stimulation were highly specific for epitopes on beef insulin, whereas pork insulin stimulation generated T cells that recognized determinants shared with beef insulin. Included among TCL reactive with pork insulin is one line (P2/9) that is autoreactive with human insulin. Antigen-presenting cells of known HLA type and monoclonal antibodies directed at class II major histocompatibility complex antigens were used to confirm the role of HLA-DR in restricting the response of insulin immune T cells. No preference or determinant selection within the donor's haplotypes was identified because either DR1 or DRw6 antigen-presenting cells could present the A loop of beef insulin. A TCL that recognized the A loop of beef insulin in association with DR1 was also alloreactive to HLA DR3, or a molecule closely linked to it, in the absence of insulin. A second T cell clone with insulin specificity and alloreactivity was also derived by allo stimulation of the donor's cells with DR3+ cells. When tested with a series of DR3+ stimulator cells, the alloreactivity was directed at diabetes-associated haplotypes. These data show that the T cell repertoire for insulin of a single diabetic donor encompasses that of multiple inbred animal strains and includes fine specificity for one to two amino acids, recognition of autologous insulin, and cross-reactivity with an allogeneic major histocompatibility complex antigen.

Animals↗
PubMed1987

Fluorescence immunohistochemical detection of hypoxic cells in spheroids and tumours.

Polyclonal antibodies have been raised in rabbits to a haemocyanin adduct of a reductively-activated, fluorinated analogue of misonidazole. Fluorescence immunohistochemical studies show that the polyclonal antibodies bind to spheroid sections and tumour sections in patterns similar to those revealed by autoradiographic studies with a tritium-labelled derivative of the fluorinated misonidazole analogue.

Animals↗
PubMed1986

Computed tomographic assessment of radiation induced damage in the lung of normal and WR 2721 protected LAF1 mice.

LAF1 mice were irradiated with single, graded doses of X rays to the thorax in the range of 0 to 14 Gy unprotected, or 0 to 18 Gy after injecting the radioprotective aminothiol compound, WR 2721. Computed tomographic (CT) scanning of the thorax was performed at intervals for a period of 42 weeks after irradiation. The gravimetric density was determined for both left and right lungs by averaging the CT numerical data within lung slices traced on a magnified video image of the thorax. Significant elevations in CT density occurred at post-irradiation times corresponding to pneumonitis and late phase, as evidenced by the pneumopathic decline in survival. The threshold dose yielding a significant increase in CT density in the pneumonitis phase was 11 Gy, a dose at which only 3% of the animals died. A single peak of increased CT density was observed for the pneumonitis phase for unprotected animals, whereas a transient return of CT density toward control values at 21-22 weeks produced two peaks from the WR 2721 treated group. The CT density of lung increased in a stepwise manner in the dose range of 11-14 Gy. For the isoeffect dose that produced equal animal survival (14 Gy and 18 Gy + WR 2721), the lung density increased by approximately 27% over control values for both treatments, suggesting that CT density is related to survival. Periodic computed tomographic analysis of the lungs of patients sustaining radiotherapy to large pulmonary fields may be of value in assessing the degree and progression of pulmonary complications.

Amifostine↗
PubMed1986

Clustered cases of paravaccinia in milkers.

Clustered cases of a disease in men and cows firstly diagnosed as cowpox has been described. Clinical manifestation, epidemiology and laboratory diagnostics are presented. Virological and serological investigations of the specimens taken from sick persons and animals proved paravaccinia virus (genus Parapoxvirus) to be etiological agent of the illness. Cowpox virus (genus Orthopoxvirus) greatly differs from the latter by phenotypical markers. The disease in humans is called Milkers' modules, in cows--pseudocowpox. The disease is mostly benign and this is a reason why it does not attract sufficient attention of health personnel. Nevertheless this infection can cause a certain economical loss.

Adolescent↗
PubMed1985

Genetic control of immune response to staphylococcal nuclease. XII: Analysis of nuclease antigenic determinants using anti-nuclease monoclonal antibodies.

SJL mice, which are high responders to Staphylococcal nuclease (nuclease), were immunized and used to produce hybridoma cell lines secreting anti-nuclease monoclonal antibodies (mAb). Ten stable clones were derived from a single fusion. Seven of these produced antibodies of the IgG1, kappa isotype and were more precisely characterized for antigenic specificity. Only one hybridoma cell line (54-10-4) produced anti-nuclease antibodies capable of inhibiting enzymatic activity of nuclease. Binding inhibition analyses strongly suggest that the other monoclonal antibodies, which failed to inhibit nuclease activity detect two different antigenic regions, or epitopes, of the molecule: epitope cluster 1 domain is defined by hybridomas 54-2-7, 54-5-2, 54-9-8, and 54-10-8; epitope cluster 2 by 54-5-1 and 54-1-9. Because of its capacity to inhibit nuclease enzymatic activity mAb 54-10-4 was considered specific for a third epitope of the nuclease molecule called epitope 3. Binding studies of these monoclonal antibodies were extended to peptide fragments of the nuclease molecule in order to examine possible cross-reactions with such fragments, as has previously been reported for antibodies purified from polyclonal antisera. Monoclonal antibodies specific for epitope cluster 1 on the native molecule also bound to the fragments 1-126 and 49-149 but failed to bind to fragment 99-149, suggesting that the corresponding epitope(s) is determined by amino acids localized between residues 49 and 99. The epitope clusters 2 and 3 appeared to be expressed only on the native molecule. Monoclonal antibodies of different clusters exhibited very different migration patterns on isoelectric focusing while monoclonal antibodies of the same cluster were indistinguishable, which suggests that they may have originated from the same B cell precursor. Taken together these data suggest that this panel of monoclonal antibodies detects at least three distinct epitopes of the nuclease molecule, one of which could be involved in the determination of the enzymatic site.

Animals↗
PubMed1985

An immunochemical probe for 8,5'-cycloadenosine-5'-monophosphate and its deoxy analog in irradiated nucleic acids.

Polyclonal antisera specific for 8,5'-cycloadenosine-5'-monophosphate (8,5'-cyclo-AMP) or its deoxy analog (8,5'-cyclo-dAMP) were elicited by immunizing rabbits with a conjugate prepared by the method of Johnston et al. [Biochemistry 22, 3453-3460 (1983)]. A competitive enzyme-linked immunosorbent assay (ELISA) was developed and used to detect the formation of these products in irradiated solutions of polyadenylic acid [poly(A)] or DNA which were saturated with nitrous oxide, nitrogen, or oxygen. The 8,5'-cyclo-AMP or 8,5'-cyclo-dAMP moieties could be detected in poly(A) at 1.0 krad and in native DNA at 10 krad, respectively. The yield of 8,5'-cyclo-dAMP was found to be two to three times higher in irradiated double-stranded DNA than in single-stranded DNA. The hydroxyl radical appears to initiate 8,5'-cyclonucleotide formation in irradiated nucleic acids, as demonstrated by the inhibition of 8,5'-cyclo-AMP formation in irradiated poly(A) by dimethyl sulfoxide. However, irradiation under nitrous oxide, particularly at low doses, does not lead to the expected increases in the yield of the 8,5'-cyclonucleotide.

Adenosine Monophosphate↗
PubMed1985

[Characteristics of a cell line obtained from a human hypernephroma].

A stable tumorigenic cell line has been obtained from the human hypernephrome. The culture grew as monolayers of large polygonal cells with foamy cytoplasm and 1-2 nucleoli. The cells formed monolayer after the third day after planting into flask and then formed multilayered growth. The electron microscopic studies of hypernephrome sections revealed the dark and light cells. The hypernephrome cells have characteristics of tumor cells. On the 200th passage the cells are aneuploid with a model number of 62 chromosomes; an acrocentric chromosome in D group is noted as a marker one. The hypernephrome cells are free of Mycoplasma contamination, able to support the several virus replication.

Animals↗
PubMed1983

Action of some hydroxyl radical scavengers on radiation-induced haemolysis.

Human and bovine erythrocytes (RBCs) from peripheral blood were gamma-irradiated in vitro to a dose of 500 Gy in the presence of three efficient hydroxyl radical (OH) scavengers: ethanol, ethylene glycol and dimethyl sulphoxide (DMSO). Bovine erythrocytes were strongly protected from radiation induced haemolysis by each of the three scavengers over a concentration range from 10(-4) to 10(-2) molar, presumably as a result of OH scavenging. Human cells were protected as efficiently as bovine RBCs by ethanol and ethylene glycol over the same concentration range, however DMSO failed to protect human cells from haemolysis over a six-decade concentration range up to one molar. Exogenously supplied vitamin E (alpha-tocopherol) protected human RBCs from haemolytic effects of 500 Gy radiation in a dose-dependent fashion; however, bovine cells were not protected over the same concentration range. These preliminary results support evidence from model membrane systems suggesting that secondary radicals of DMSO generated during radiation may be of sufficient reactivity to initiate lipid peroxidation and are suggestive of species differences in the protection of biological membranes from oxidative stress.

Animals↗