Interpretation of survival-curve data for Chinese hamster cells, line V-79 using the multi-target, multi-target with initial slope, and alpha, beta equations.
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Biomedical subjects
Publications and source records attributed to B C Millar.
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Three biradical nitroxyl compounds have been shown to modify the slope and shoulder region of the hypoxic cell survival curve of V79 cells to different degrees. The amount of sensitization produced by these compounds is dependent on the cell-drug contact time at both 20 degree and 37 degree C whereas sensitization by the monoradical nitroxyl, NPPN, is independent of these factors. The results suggest that biradicals may modify cellular biochemistry in such a way as to change the repair capacity of the cells. This could be responsible for changes in the shape of survival curves when cell-drug contact times are increased.
The biradical nitroxyl, Ro.03-6061, sensitizes two lines of mouse L cell to ionizing radiation when the cells are rendered hypoxic. Although the biradical reduces the Do value of the hypoxic cell-survival curve in each instance, it has no significant effect on the shoulder region. A hybrid line produced from these two strains is more radioresistant than either parent. In this instance, the biradical suppresses the shoulder region of the hypoxic cell-survival curve, but has no effect on the Do value. In a second system, the biradical selectivity sensitizes hypoxic cells of a diploid and a tetraploid clone of Syrian hamster cells (BHK21/C15). The survival-curve characteristics of both clones are similar. The biradical reduces the Do value but does not significantly change the shoulder region of the hypoxic cell-survival curve. An aneuploid line sub-cultured from the tetraploid clone is much more resistant to radiation. In this instance, there is a decrease in the Do value of hypoxic cells in the presence of the biradical, but the extrapolation number is increased to a value similar to that for cells irradiated in air.
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A study of 32 patients receiving cyclophosphamide (CY) and verapamil (VER) in addition to the drug combination vincristine, adriamycin and methyl prednisolone (VAMP) was made in which the clinical response and growth of clonogenic myeloma cells (MY-CFUc) from bone marrow aspirates were compared. At presentation, MY-CFUc were grown from 72 per cent (23/32) of the patients. After treatment with CY-VAMP or VERCY-VAMP, MY-CFUc were grown from 25 per cent (8/32) of patients of whom only 50 per cent responded clinically. The overall clinical response rate for patients receiving CY-VAMP and VERCY-VAMP was 64 per cent (9/14) and 72 per cent (13/18) respectively of whom 14 per cent in each group achieved complete remission. There was no concomitant increase in normal tissue toxicity as measured by granulocyte-macrophage colony (GM-CFUc) formation. Comparison of these data with our previous study of patients receiving VAMP alone, suggests that the addition of CY to the regimen may increase the tumour cell kill. Further clinical studies will determine whether there is a significant increase in the complete remission rate.
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BACKGROUND: Patients with cystic fibrosis (CF) are at high risk from atypical mycobacterial infections. There have been few attempts to delineate the intensity of mycobacterial infection in CF patients in Ireland. AIMS: To examine the incidence of mycobacterial DNA in an archived collection of genomic DNA extracted from the sputa of CF patients within the Northern Ireland population. METHODS: One hundred and eighty-two CF patients (66 adults and 116 children) were examined for the presence of mycobacterial DNA in their sputum by a genus specific PCR assay based on 16S rRNA, followed by direct automated sequencing of the PCR amplicons. RESULTS: One of 116 (0.9%) children and 2 of 66 adults were positive. Sequence identity revealed Mycobacterium xenopi in the paediatric patient and M. xenopi and M. chelonei in the two adult patients. False-positive results occurred in 11 patients (four adults), mainly due to Corynebacterium spp. CONCLUSIONS: There was a low prevalence of Mycobacterium spp in the CF patient population. All PCR positive results should be confirmed by direct automated sequencing and an alternative specific assay employed. Enhanced molecular screening will contribute in understanding their role as opportunistic pathogens in patients with worsening lung function.
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BACKGROUND: Cryptosporidium parvum is the most common of the protozoal pathogens associated with gastrointestinal disease in Northern Ireland. Genotyping techniques are valuable in helping to elucidate sources and modes of transmission of this parasite. There have been no reports on the prevalence of genotypes in Northern Ireland, mainly due to a lack of discriminatory genotyping techniques, which recently have become available. AIM: To investigate the genotype of C. parvum oocysts isolated from human faeces in sporadic cases of cryptosporidiosis in Northern Ireland. METHODS: Thirty-nine isolates of C. parvum, representing 79.6% of the total 1998 laboratory reports for the Eastern Health and Social Services Board, were investigated. Following DNA extraction from oocysts the thrombospondin-related adhesive protein 2 (TRAP-C2) locus was amplified by polymerase chain reaction (PCR) and subsequently sequenced. RESULTS: The majority of isolates (87.2%) were classified as bovine genotype II with the remainder (12.8%) being the human genotype I. CONCLUSIONS: There is a high prevalence of the bovine genotype II parasite in sporadic cases around the greater Belfast area. Epidemiologically, this suggests that the most frequent mode of transmission may be from animals to humans, but does not suggest a high proportion of human to human spread.
BACKGROUND: No study has compared Campylobacter isolates from a human source in Northern Ireland over an extended period of time. AIM: To investigate the clonal stability of thermophilic campylobacters isolated from acute bacterial enteritis in Northern Ireland from 1992 to 1999. METHODS: Human isolates (n=272), originating from faeces, were characterised at the sub-species level using enterobacterial repetitive intergenic consensus sequence (ERIC2)--random amplification of polymorphic DNA (RAPD) typing. RESULTS: Thirteen genotypes were identified where three types, namely ERIC A, ERIC C and ERIC I, accounted for 28.3%, 14.3% and 13.6%, respectively. There were no significant associations (p>0.05) between sex, age groupings and year of isolation and ERIC2 genotype, with the exception of ERIC D, which showed a significant decline in isolation with time (p=0.019). CONCLUSIONS: ERIC genotypes were stable over this period, except genotype C which was lost during this time. Previous molecular typing methods showed campylobacters to be heterogeneous but this study suggests that the local human Campylobacter population consists of several common and stable genotypes. This study has established a database of local ERIC2 genotypes. This methodology may allow the establishment of an all-island database of clinical campylobacters that would be valuable in reducing human campylobacteriosis in Ireland.
Fungal endocarditis in the neonate and pediatric population is associated with a high degree of morbidity and mortality, with diagnosis in the majority of cases made postmortem. The aim of this article is to review published literature on fungal endocarditis in children and to discuss the incidence due to a wide range of fungi, primarily Aspergillus and Candida species, as well as its diagnosis, including the importance of molecular-based approaches and various treatment regimes.
Overnight exposure of Chinese hamster cells, V-79-753B, to certain non-steroidal anti-inflammatory agents (NSAIA) including indomethacin (5 x 10(-5)M), benoxaprofen (5 x 10(-5)M) or aspirin (10(-4)M) protected against misonidazole-induced toxicity both in air and in hypoxia at 37 degrees C. In no instance was the radiosensitivity of cells affected by these treatments nor was there any effect on the amount of acute hypoxic cell radiosensitization produced by 1.0 mM misonidazole. There was no protection against misonidazole-induced toxicity when cells were pretreated with theophylline (10(-3)M). Protection against misonidazole-induced toxicity by benoxaprofen was not reversed by the addition of 1 microgram/ml prostaglandin E1 or F1 alpha. The results are discussed in terms of possible mechanisms.
A wild-type isolate with similar morphological and phenotypic properties to Taylorella equigenitalis, the causative bacterial agent of contagious equine metritis (CEM), was referred for molecular identification by PCR amplification of the 16S rRNA gene. A species-specific PCR failed to yield a product compatible with that of T. equigenitalis. The direct sequencing of the universal 16S rRNA PCR amplicon suggested the presence of a Bacteroides sp., probably Bacteroides ureolyticus, with no consequent effects on the movement and transportation of the animal. Adoption of such a molecular means of identification through sequencing may aid in the identification of the atypical forms of Taylorella equigenitalis, as recently described, as well as differentiating this species from Taylorella asinigenitalis.