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Biomedical subjects

J Hilton

Publications and source records attributed to J Hilton.

At least 55 records · Page 3Linked to original sources
PubMed1996

Graft-versus-myeloma effect: proof of principle.

The presence of a graft-versus-tumor effect has been well established in leukemia but not in multiple myeloma. A 40-year-old patient with myeloma refractory to standard chemotherapy and autologous transplantation received a matched unrelated T-cell-depleted transplant after conditioning with fractionated total-body irradiation, thiotepa, and cyclophosphamide. This procedure resulted in a transient and incomplete response with evidence of rapidly progressive disease within 2.5 months posttransplantation. The patient then received a small number of donor peripheral blood (PB) mononuclear cells (CD3 cells 1.2 x 10(6)/kg) without any further cytotoxic therapy. A complete remission was attained, lasting now for more than 14 months. The procedure was associated with severe acute and subsequently limited chronic graft-versus-host disease (GVHD). This report provides the first direct evidence of a graft-versus-myeloma effect after allogenic transplantation.

Adult↗
PubMed1996

Influence of dibutyl phthalate on dermal sensitization to fluorescein isothiocyanate.

What limited evidence there is indicates that the formulation in which a chemical allergen is encountered on the skin can have a marked impact upon the induction of cutaneous immune responses and the subsequent development of contact sensitization. The purpose of the present investigations was to examine further this phenomenon by analysis of the influence of dibutyl phthalate (DBP) on dermal sensitization to fluorescein isothiocyanate (FITC), a skin sensitizing fluorochrome. Addition of DBP augmented very substantially, in a dose-dependent fashion, the ability of topically applied FITC to stimulate proliferative responses in mice by draining lymph node cells (LNC), a correlate of skin sensitizing potential. Under these conditions, exposure of mice to DBP alone failed to elicit significant LNC responses. The influence of DBP on the accumulation of dendritic cells (DC) induced by FITC was examined also. Although 10% DBP had little effect on the numbers of DC found within draining nodes 18 hr following exposure of mice to FITC, the phthalate did result in a very substantial increase in the frequency of lymph node DC bearing detectable antigen (FITC+ DC). Furthermore, in the presence of DBP the median amount of FITC associated with antigen-bearing DC was higher. In vitro skin absorption studies indicated that DBP was associated with a small increase in percutaneous absorption of FITC. Collectively these data demonstrate that the vehicle formulation can exert a marked influence on dermal sensitization and that one mechanism which may be relevant is the increased acquisition of antigen by DC, associated possibly with altered penetration of the allergen into or through the skin.

Administration, Topical↗
PubMed1996

Experimental assessment of the sensitizing properties of formaldehyde.

Formaldehyde causes upper respiratory tract irritation and has been reported in some investigations to be a cause of occupational allergic asthma. The data are equivocal, however, and it has proved difficult to confirm that exposure to formaldehyde induces respiratory sensitization or provokes the production of specific immunoglobulin E (IgE) antibody. In this study the sensitizing properties of formaldehyde were examined experimentally. This chemical elicited strong positive responses in three independent methods for the prospective identification of contact sensitizing chemicals-the guinea pig maximization test, the occluded patch test of Buehler and the murine local lymph node assay. In contrast, in a novel predictive test method for assessment of respiratory sensitization potential-the mouse IgE test-formaldehyde at the same test concentrations was negative. Furthermore, formaldehyde induced in mice a pattern of cytokine secretion by draining lymph node cells inconsistent with the stimulation of IgE antibody responses or respiratory sensitization. These data indicate that, although formaldehyde is a potent contact allergen, it lacks a significant potential to cause sensitization of the respiratory tract.

Administration, Topical↗
PubMed1996

Predictive testing for respiratory sensitization in the mouse.

Attempts to develop predictive test methods for the identification of chemical respiratory allergens have to date focused almost exclusively on the guinea pig. In recent years there has, however, been a growing interest in the mouse as a model for examination of sensitization potential. In this article two alternative approaches to the toxicological investigation of respiratory sensitization are described. Both are based on an understanding of the nature of immune responses induced in mice by chemical allergens. The mouse IgE test seeks to identify chemicals capable of causing allergic sensitization of the respiratory tract as a function of induced increases in the serum concentration of IgE. The second approach, cytokine fingerprinting, makes use of the observation that chemical allergens of different types provoke in mice qualitatively divergent immune responses characterized by discrete cytokine secretion profiles.

Allergens↗
PubMed1996

Dichloronitrobenzene: a reappraisal of its skin sensitization potential.

Unlike the closely related chemical dinitrochlorobenzene (DNCB), which is a very strong contact allergen, dichloronitrobenzene (DCNB) has been widely regarded as a non-allergen and, as such, a useful control for its strongly sensitizing counterpart. Nevertheless, it is still an organic chemical species readily capable of penetrating skin and, rather than being regarded as completely inert, it has even been suggested to react with the immune system in such a way that it induces specific tolerance to its chemical structure. We investigated whether DCNB was in reality a non-allergen, or rather merely a weak contact sensitizer. In both a rigorously conducted guinea pig maximization test and in a modified murine local lymph node assay, DCNB was demonstrated to possess weak sensitizing activity. On this basis, DCNB cannot be regarded as inert with respect to contact allergic potential, and is therefore inappropriate as a negative control in studies of skin sensitization.

Animals↗
PubMed1996

Minimizing graft rejection in allogeneic T cell-depleted bone marrow transplantation.

Between October 1991 and May 1994, 42 patients were treated with cyclophosphamide, thiotepa, and total body irradiation followed by an allogeneic transplantation of marrow depleted of T cells with soybean agglutinin and E-rosetting. Patients included in this study had acute myelogenous leukemia (13), chronic myelogenous leukemia (12), acute lymphocytic leukemia (nine), Hodgkin's disease or non-Hodgkin's lymphoma (four), multiple myeloma (three), or myelodysplastic syndrome (one). The mean age was 34 (range 8 to 51 years). Nineteen patients had a matched sibling donor and 18 received marrow from 6/6 matched unrelated donors while five received transplants from unrelated donors disparate at one DR locus (5/6 match). Time to granulocyte engraftment (AGC > or = 500/mm3) occurred at a mean of 16.5 days for related and 11.4 days for unrelated transplant recipients, and was related to the increased use of G-CSF in the unrelated population. There was no correlation with number of mononuclear cells, T cells, or CD34-positive cells infused, the rate of engraftment or the incidence of transplant complications. Multivariate analysis determined that G-CSF administration and a diagnosis other than ALL were the only factors associated with a faster rate of engraftment. Patients receiving unrelated donor transplants, those with ALL, or those who had a low T cell number infused (< or = 8.0 x 10(3) cells/kg) experienced delayed hospital discharge. The regimen resulted in excellent rates of engraftment (95.2%) with only one failure to engraft and one graft rejection. The incidence of grade III-IV acute graft-versus-host disease was 0% with sibling and 26.1% with unrelated donors. There were no cases of veno-occlusive disease. Fifty percent of patients are alive with a mean follow-up of 26.4 months. We conclude that this regimen is well tolerated and results in excellent engraftment with a low incidence of severe graft-versus-host disease and few therapy-related toxicities.

Adolescent↗
PubMed1995

An international evaluation of the murine local lymph node assay and comparison of modified procedures.

The murine local lymph node assay is a predictive test for the identification of skin-sensitizing chemicals. The method has been the subject both of national inter-laboratory studies and of extensive comparisons with guinea pig tests. In the investigations reported here, the local lymph node assay has been evaluated further in the context of an international study comprising five independent laboratories. In addition, the influence of minor modifications to the standard assay procedure on the performance of the test has been examined. The modified procedures investigated were exposure of mice for 4 rather than 3 consecutive days, excision of lymph nodes 4 rather than 5 days after the initiation of exposure and the use of an alternative isotope. All five laboratories, irrespective of whether the standard or a modified protocol was used, were able to identify accurately, and with comparable sensitivity, potassium dichromate and 2,4-dinitrochlorobenzene as skin sensitizers. Using standard criteria, none of the laboratories recorded positive responses with methyl salicylate, a non-sensitizer. In the standard protocol, lymph nodes are pooled for each experimental group and the vigor of responses measured as a stimulation index relative to vehicle controls. A stimulation index of 3 or greater is considered to indicate skin-sensitizing potential. One further modification adopted by three of the laboratories was to analyze nodes from individual animals and, thereby, permit statistical evaluation. This allowed a direct comparison of statistical significance with the conventional stimulation index as criteria for a positive response. The data indicate that, while statistical evaluation may provide, in some instances, for small increases in sensitivity, this may be at the expense of some loss of selectivity. There are, however, insufficient data presently to draw firm conclusions regarding the relative value of statistical analysis. These studies demonstrate that the local lymph node assay is sufficiently robust to accommodate minor procedural and technical modifications without material changes in test performance.

Analysis of Variance↗
PubMed1995

BCR-ABL-mediated inhibition of apoptosis with delay of G2/M transition after DNA damage: a mechanism of resistance to multiple anticancer agents.

A critical determinant of the efficacy of antineoplastic therapy is the response of malignant cells to DNA damage induced by anticancer agents. The p53 tumor-suppressor gene is a critical component of two distinct cellular responses to DNA damage, the induction of a reversible arrest at the G1/S cell cycle checkpoint, and the activation of apoptosis, a genetic program of autonomous cell death. Expression of the BCR-ABL chimeric gene produced by a balanced translocation in chronic myeloid leukemia, confers resistance to multiple genotoxic anticancer agents. BCR-ABL expression inhibits the apoptotic response to DNA damage without altering either the p53-dependent WAF1/CIP1-mediated G1 arrest or DNA repair. BCR-ABL-mediated inhibition of DNA damage-induced apoptosis is associated with a prolongation of cell cycle arrest at the G2/M restriction point; the delay of G2/M transition may allow time to repair and complete DNA replication and chromosomal segregation, thereby preventing a mitotic catastrophe. The inherent resistance of human cancers to genotoxic agents may result not only by the loss or inactivation of the wild-type p53 gene, but also by genetic alterations such as BCR-ABL that can delay G2/M transition after DNA damage.

Antineoplastic Agents↗
PubMed1995

Effect of stereochemistry on the oxidative metabolism of the cyclophosphamide metabolite aldophosphamide.

31P NMR and cell perfusion techniques were used to investigate the conversion of the individual enantiomers of aldophosphamide (AP) to carboxyphosphamide (CBP) as catalyzed by aldehyde dehydrogenase in human erythroleukemia K562 cells. R- and S-cyclophosphamides (CPs) were treated with ozone and hydrogen peroxide to yield Rp- and Sp-cis-4-hydroperoxycyclophosphamides (Rp- and Sp-cis-4-HO2-CP); reduction of each hydroperoxide gave the corresponding enantiomer of AP [along with its tautomer 4-hydroxycyclophosphamide (4-HO-CP)]. In separate experiments, K562 cells embedded in agarose gel threads were perfused at pH 7.4, 21 +/- 1 degrees, with solutions of 1.4 mM Rp- and Sp-4-HO-CP/AP, both with and without added mesna (an acrolein scavenger). A comparison of the 31P NMR spectral data derived from the experiments revealed little statistical difference (+/- 10-20% error limits) in the normalized intensities of the CBP peaks arising from the individual AP enantiomers [with added mesna, the ratio Rp-CBP:Sp-CBP was 1.00:1.24 +/- 0.13 (average deviation); without mesna, the same ratio was 1.00:1.35]. Using conventional methods for evaluating the in vitro drug toxicities, CP-resistant L1210 cells were treated in separate experiments with Rp- and Sp-cis-4-HO2-CP; there were no significant differences between the toxicities exhibited by the stereoisomers.

Aldehyde Dehydrogenase↗
PubMed1995

Assessment of aldehyde dehydrogenase in viable cells.

Cytosolic aldehyde dehydrogenase (ALDH), an enzyme responsible for oxidizing intracellular aldehydes, has an important role in ethanol, vitamin A, and cyclophosphamide metabolism. High expression of this enzyme in primitive stem cells from multiple tissues, including bone marrow and intestine, appears to be an important mechanism by which these cells are resistant to cyclophosphamide. However, although hematopoietic stem cells (HSC) express high levels of cytosolic ALDH, isolating viable HSC by their ALDH expression has not been possible because ALDH is an intracellular protein. We found that a fluorescent aldehyde, dansyl aminoacetaldehyde (DAAA), could be used in flow cytometry experiments to isolate viable mouse and human cells based on their ALDH content. The level of dansyl fluorescence exhibited by cells after incubation with DAAA paralleled cytosolic ALDH levels determined by Western blotting and the sensitivity of the cells to cyclophosphamide. Moreover, DAAA appeared to be a more sensitive means of assessing cytosolic ALDH levels than Western blotting. Bone marrow progenitors treated with DAAA proliferated normally. Furthermore, marrow cells expressing high levels of dansyl fluorescence after incubation with DAAA were enriched for hematopoietic progenitors. The ability to isolate viable cells that express high levels of cytosolic ALDH could be an important component of methodology for identifying and purifying HSC and for studying cyclophosphamide-resistant tumor cell populations.

Aldehyde Dehydrogenase↗
PubMed1995

Molecular pharmacology of hepsulfam, NSC 3296801: identification of alkylated nucleosides, alkylation site, and site of DNA cross-linking.

We have determined that hepsulfam, in common with its structural homologue busulfan, alkylates both free guanosine and GMP in DNA at the 7 nitrogen. Mass spectral analysis of the products of the reaction of hepsulfam with guanosine has identified the mono- and bis-alkylated guanosine adducts. UV spectrophotometry and mass spectrometry were used to confirm that alkylation occurred at the 7 nitrogen by following the formation of the formamidopyrimidyl form of the hepsulfam-guanosine adduct at high pH. We have also isolated and identified 1-guanyl,7-hydroxyheptane, 1-guanyl,7-sulfamylheptane, and 1,7-bis(guanyl)heptane from in vitro reaction mixtures of hepsulfam and calf thymus DNA. We have isolated bis-(7-formamidopyrimidyldeoxyguanosinyl)-heptane from an enzymatic digest of DNA treated with hepsulfam. Finally, we have found that hepsulfam forms interstrand cross-links at 5'-GXC-3' sites in model oligonucleotides.

Alkylation↗
PubMed1995

Resonance Raman spectroscopic characterization of the molybdopterin active site of DMSO reductase.

Resonance Raman spectra are compared for Rhodobacter sphaeroides dimethyl sulfoxide reductase, an enzyme containing a molybdopterin cofactor, and two model compounds, I and II, which have pterin and quinoxaline, respectively, attached to a Cp2Mo[IV]-dithiolene chelate [Cp = cyclopentadienyl]. The effect of 34S incorporation was also determined. Several bands in the 200-500 cm-1 region show remarkably similar patterns of frequencies and isotope shifts between protein and models: a band at 351 cm-1 shifts 6-8 cm-1, and bands at lower and higher frequencies show smaller shifts upon 34S substitution. A normal coordinate analysis on II indicates the 351 cm-1 mode to be the symmetric Mo-S[dithiolene] stretch and the remaining low-frequency modes to contain contributions from deformations of the quinoxaline ring as well as from Mo-S stretching. The similarity in the low-frequency spectra between the model compounds and the enzyme strongly supports a dithiolene chelate as the mode of Mo-pterin interaction in the cofactor. Resonance enhancement of both high- and low-frequency quinoxaline or pterin modes is observed for both model compounds, implicating the heterocyclic rings as part of the electronic system involved in the Mo-dithiolene charge transfer transitions. RR spectra of 6-methylpterin and biopterin are reported and used to identify the pterin and quinoxaline high-frequency bands in the model compound spectra. The dithiolene C = C stretch is tentatively assigned to bands at 1506 cm-1 in I and 1515 cm-1 in II.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding Sites↗
PubMed1995

Differences in diaphragm fiber types in SIDS infants.

The diaphragm is a vital respiratory muscle in the sleeping infant. Any changes in diaphragm fiber type number or size could represent either a primary developmental delay or a secondary reaction to increased workload, and could give a clue as to the pathogenesis of sudden infant death syndrome (SIDS). We therefore quantitated by point counting on ATPase histochemistry the numbers and areas of type 1 and 2 fibers in the diaphragm, external intercostal and psoas muscles of 37 SIDS and 20 control infants. The amount of slow, fast and fetal myosin in the diaphragm and psoas muscles was measured by electrophoresis to check the ATPase quantitation. There were fewer type 1 fibers in SIDS (median 30.0%) compared with control (median 40.0%) infants (p < 0.02), whereas the diameter of type 1 fibers in SIDS (median 33.9 microns) was larger than in control (median 30.3 microns) infants (p < 0.007). The total cross-sectional area occupied by type 1 and 2 fibers was similar in both groups. No changes were found in the external intercostal or psoas. The amount of slow and fast myosins correlated well with type 1 and type 2 fibers, respectively. The finding of fewer type 1 (fatigue-resistant) fibers of large diameter in SIDS diaphragms suggests that differences in muscle fiber types may predispose these infants to diaphragm fatigue and respiratory failure.

Diaphragm↗
PubMed1995

Brain stem nuclei in sudden infant death syndrome (SIDS): volumes, neuronal numbers and positions.

It has been suggested that the defect underlying the sudden infant death syndrome (SIDS) lies in brain stem nuclei involved in cardiac and respiratory function. However, most studies have not used rigorous quantitative techniques to assess brain stem nuclear volumes and neuronal numbers. We have measured the volume, neuronal numbers and position of brain stem nuclei in 11 SIDS and 11 aged-matched control infants. Using serial sagittal sections, nuclei involved in maintaining airway patency (hypoglossal, ambiguus and retroambiguus), heart rate (dorsal vagal) and generation of respiratory rhythm (ambiguus and dorsal vagal) were studied. No significant differences were found in nuclear volume increase with age, total neuronal number or nuclear position between SIDS and control cases. These findings support the hypothesis that the nervous system in SIDS may be normal until the final event that kills these infants.

Age Factors↗
PubMed1995

Streptozotocin: inherent but not expressed skin sensitizing activity.

We have shown previously that the alkylating agent streptozotocin (STZ) fails to induce responses in the murine local lymph node assay, a predictive test for contact sensitizing potential. In the present study, we demonstrate that this same compound when injected intradermally into the ears of mice provokes a dose-dependent induction of draining lymph node cell proliferation. These data indicate that the intrinsic immunogenicity of STZ is not displayed in the local lymph node assay under conditions of conventional topical exposure, secondary to a failure to cross the lipophilic stratum corneum in sufficient quantities. This finding has implications for the discernment of structure-activity relationships in contact sensitization.

Animals↗
PubMed1994

Interstitial taxol delivered from a biodegradable polymer implant against experimental malignant glioma.

Taxol is a novel antitumor agent with demonstrated efficacy against ovarian, breast, and non-small cell lung cancers in Phase II clinical trials, but which has been shown not to cross the blood-brain barrier. To adapt taxol as a therapy for brain tumors, we have incorporated it into a biodegradable polyanhydride matrix for intracranial implantation and evaluated this formulation in a rat model of malignant glioma. Fischer 344 rats bearing intracranial 9L glioma tumors were treated with 10 mg poly[bis(p-carboxyphenoxy)propane-sebacic acid] (20:80) copolymer discs, containing 20-40% taxol by weight, 5 days after tumor implantation. The taxol-loaded polymers doubled (38 days, 40% taxol loading, P < 0.02) to tripled (61.5 days, 20% taxol loading, P < 0.001) the median survival of rats bearing tumor relative to control rats (19.5 days). Drug loadings of 20-40% taxol by weight released intact taxol for up to 1000 h in vitro. In rats followed up to 30 days postimplant, the polymer maintained a taxol concentration of 75-125 ng taxol/mg brain tissue (100-150 microM taxol) within a 1-3-mm radius of the disc. At points more distant from the disc (up to 8 mm away, the size limit of the rat brain), the polymer maintained a taxol concentration of greater than 4 ng taxol/mg brain tissue (5 microM). We conclude that taxol shows promise as a therapy for malignant glioma when delivered interstitially from a biodegradable polymer.

Animals↗