• Less G(2) arrest in irradiated cells of breast cancer patients than in female controls: a contribution to their enhanced chromosomal radiosensitivity?

      Scott, David; Spreadborough, Anne R; Roberts, Stephen A; Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester M20 4BX, UK. dscott@picr.man.ac.uk (2003-06)
      PURPOSE: To determine if the efficacy of G(2) checkpoint control (measured as the degree of mitotic inhibition) was reduced in breast cancer patients (n=129) compared with healthy controls (n=105) after exposure of lymphocytes to X-rays. We had previously shown that the average level of radiation-induced chromosome damage was higher in G(2) lymphocytes of these patients than in the controls, and it was proposed that this was a marker of low penetrance predisposition to cancer. MATERIALS AND METHODS: Proliferating lymphocytes were X-irradiated (50 cGy) and sampled at 90 min post-irradiation, which was the time of maximum mitotic inhibition of G(2) cells, expressed as the extent of reduction in the mitotic index in irradiated compared with unirradiated cells. RESULTS: Repeated measurements on 28 controls showed that there were reproducible differences in mitotic inhibition between individuals. Inhibition was significantly greater in female than in male controls (p=0.014), but less in patients than in female controls (p=0.009). There was a weak inverse correlation between the extent of inhibition and the amount of chromosome damage in all females (r=-0.15, p=0.043). CONCLUSIONS: The lesser mitotic inhibition in patients than in female controls might contribute to their greater mean G(2) chromosomal radiosensitivity. However, this hypothesis is not easily reconciled with other observations that (1) the significant difference in inhibition between the sexes in controls was not accompanied by any gender difference in radiosensitivity and (2) there was an inverse correlation between inhibition and age in controls, yet no age-related increase in radiosensitivity. There might, therefore, be no causal relationship between G(2) mitotic inhibition and chromosomal radiosensitivity.
    • Letter to the editor in response to 'When to apply sunscreen: a consensus statement for Australia and New Zealand'

      Baldwin, L; Olsen, CM; Gordon, L; Green, Adèle C; Aitken, J; Neale, R; Whiteman, D; Janda, M; Institute of Health and Biomedical Research, School of Public Health and Social Work, Faculty of Health, Queensland University of Technology, Australia (2019)
    • Leukocyte migration inhibition in human pulmonary neoplasia.

      Vose, Brent M; Kimber, I; Moore, Michael; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1977-03)
      Peripheral blood leukocytes from 65 lung cancer patients, 69 healthy donors, 33 patients with malignant disease outside the lung, and 24 patients with nonmalignant pulmonary conditions were examined for immune reactivity, as measured by the leukocyte migration inhibition assay, to antigens in homogenates of lung tumor tissue and material derived from tumor-free lung areas and a nonmalignant lung lesion. A low level of reactivity with apparent specificity was detected in the lung cancer patient group, with 17/52 showing migration inhibition with extracts of lung tumor tissue. Reactivity was also detectable in this group against homogenates of tumor-free lung and nonmalignant lung lesion. The data supported the conclusion that sensitization at least in some patients with lung cancer might be directed not only against tumor-specific antigens but also against antigens of less confined disease association.
    • Levels of the DNA adduct, N7-methyldeoxyguanosine, are associated with increased risk of failure of treatment of cervical intraepithelial neoplasia.

      Acladious, N N; Harrison, Kathryn L; Sutton, C J; Povey, Andrew C; Mandal, D; Kitchener, Henry C; Department of Genito-Urinary Medicine, Manchester Royal Infirmary, Manchester, UK. (2004-06)
      OBJECTIVE: To determine whether exposure to methylating agents was a risk factor for treatment failure in women undergoing colposcopic examination. METHODS: Nine hundred fifty-eight women attending for colposcopic examination after abnormal cervical smear test results were recruited into the study cohort. Information on demographic factors, smoking and other risk factors was obtained and a pre-treatment biopsy was taken and stored at -70 degrees C. After follow-up, cases who had treatment failure of cervical intraepithelial neoplasia (CIN) within 2 years following treatment were identified (n = 77) and matched to women with no treatment failure of CIN in this time period (controls, n = 154). DNA was extracted from the pre-treatment biopsies and levels of N7-methyl-deoxyguanosine (N7-MedG), a marker of exposure to methylating agents, were quantified as the ring-opened form of the base damage by a validated immunoslotblot assay. RESULTS: Sufficient DNA for N7-MedG analysis was extracted from 61 subjects corresponding to 20 matched case control pairs. N7-MedG was detected in cervical DNA with levels ranging from non-detected (<0.1 micromol/mol dG) to 4.83 micromol/mol dG. N7-MedG levels were significantly higher in cases (geometric mean 0.99 micromol/mol dG) than controls (0.33 micromol/mol dG; P = 0.01). There were no associations between N7-MedG levels and HPV or smoking status. Log N7-MedG content, after adjustment for HPV status at time of treatment, was found to be significantly associated with increased risk of treatment failure (OR 5.74, 95% CI 1.05-31.23). CONCLUSIONS: The association between pre-treatment levels of DNA damage induced by methylating agents and subsequent treatment failure implicates methylating agent exposure as a causative factor in treatment failure.
    • Li-Fraumeni syndrome--a molecular and clinical review.

      Varley, Jennifer; Evans, D Gareth R; Birch, Jillian M; CRC Department of Cancer Genetics, Paterson Institute for Cancer Research, Manchester, UK. (1997)
    • Light and scanning electron microscopy of the same human metaphase chromosomes.

      Harrison, Christine J; Jack, E; Allen, Terence D; Harris, R; Paterson Laboratories, Christie Hospital & Holt Radium Institute, Manchester, M20 9BX, UK. (1985-08)
      A technique has been developed to examine the same G-banded human metaphase chromosomes, first in the light microscope and then in the scanning electron microscope (SEM). A structural involvement in chromosome banding was confirmed by a positional correlation between the G-positive bands observed in the light microscope and the circumferential grooves between the quaternary coils of the metaphase chromosomes, observed in the SEM. In further support of this the regions between the grooves showed a positional relationship with the G-negative or reverse (R) bands. The examination of slightly extended metaphase chromosomes in the light microscope demonstrated that the G-banding pattern corresponded to that described by the Paris nomenclature for metaphase chromosomes. The arrangement of the circumferential grooves of the same chromosomes, observed in the SEM, was shown to relate to that described by the Paris nomenclature for prometaphase chromosomes. Therefore, using the SEM it is possible to demonstrate the details of prometaphase banding in metaphase chromosomes.
    • Limitations of immune complex measurements in colorectal disease.

      Hobbiss, J; Cooper, K M; Moore, Michael; Gowland, E; Schofield, P F; Department of Surgery, Univerity Hospital of South Manchester, Withington, Manchester (1983-08)
      Three techniques (Clq, Raji and L1210 binding assays) alleged to measure circulating immune complexes (ICs) were applied to the sera of 101 patients with colorectal disease (54 carcinoma; 23 inflammatory; 13 benign tumour and 11 miscellaneous) at the time of diagnostic or definitive surgery, and 58 healthy adult controls. Elevated levels in the pathological sera were observed by all 3 methods in order of sensitivity: Raji greater than Clq greater than L1210. However, none of them differentiated between benign, inflammatory and neoplastic conditions nor, in the case of colorectal carcinoma, was there any correlation with stage of disease. With the exception of Raji v. L1210 (r = 0.43, P less than 0.001), correlations between the various assays were poor and levels of serum carcinoembryonic antigen (CEA) did not correlate with ICs measured by any of the techniques. Indeed, the IC assays were even less discriminatory than CEA, which was elevated mainly in the serum of carcinoma patients and which was positively correlated with serum gamma-glutamyl transpeptidase (gamma GT) (r = 0.42, P less than 0.005). The data suggest that the lack of concordance between the IC assays is a reflection of heterogeneity among ICs, interfering factors present in pathological sera, or both. Thus the IC assays deployed here have neither diagnostic nor prognostic utility in colorectal disease at this time, and immunochemical characterization of the serum reactive material detected by the different assays is required.
    • The lineage commitment of haemopoietic progenitor cells.

      Cross, Michael A; Enver, T; Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital National Health Service Trust, Manchester, UK. exhmc@picr.cr.man.ac.uk (1997-10)
      Multipotent haemopoietic progenitor cells appear to be 'primed' for commitment by co-expression of a multiplicity of genes characteristic of different lineages. Lineage commitment proceeds as the consolidation of a distinct pattern of gene expression out of this milieu.
    • Lineage restriction of the RARalpha gene expression in myeloid differentiation.

      Zhu, Jun; Heyworth, Clare M; Glasow, Annegret; Huang, Qiu-Hua; Petrie, Kevin; Lanotte, Michael; Benoit, Gerard; Gallagher, Robert; Waxman, Samuel; Enver, Tariq; et al. (2001-10-15)
      To better understand the role of retinoids in myelopoiesis, expression of the retinoid receptor genes (retinoic acid receptors [RARs] and retinoid X receptors [RXRs]) were examined during differentiation of factor-dependent cell-Paterson (FDCP)-mixA4 murine progenitor cells. The major receptor expressed in undifferentiated A4 cells was RARalpha (primarily the RARalpha1 isoform). Following induction of myelomonocytic differentiation with granulocyte and granulocyte-macrophage colony-stimulating factors, a dramatic increase in RARalpha expression (particularly the RARalpha2 isoform) was seen. In contrast, expression of both RARalpha isoforms was rapidly extinguished upon induction of erythroid differentiation with erythropoeitin (EPO). A modest induction of RXRalpha expression was seen, particularly during differentiation in the myelomonocytic lineage. Low expression levels of RARgamma2 and RXRbeta remained unchanged, irrespective of differentiation pathway. Consistent with the gene expression patterns, RARalpha agonists and antagonists stimulated myelomonocytic and erythroid differentiation of FDCP-mixA4 cells, respectively. Taken together, these results suggest that erythropoiesis and granulopoiesis require diminished and enhanced RARalpha activities, respectively, which at physiological all-trans-retinoic acid (RA) concentrations may be accomplished by reciprocal effects of EPO and myelomonocytic growth factors on its expression. This hypothesis is corroborated by data showing that RA, which positively regulates RARalpha2 expression, can exert inhibitory effects on erythroid differentiation.
    • Linkage studies in a Li-Fraumeni family with increased expression of p53 protein but no germline mutation in p53.

      Birch, Jillian M; Heighway, Jim; Teare, M Dawn; Kelsey, Anna M; Hartley, Ann L; Tricker, K J; Crowther, Derek; Lane, D P; Santibanez-Koref, Mauro F; University of Manchester, CRC Paediatric & Familial Cancer Research Group, Christie Hospital, UK. (1994-12)
      We report a family with the Li-Fraumeni syndrome (LFS) in whom we have been unable to detect a mutation in the coding sequence of the p53 gene. Analysis of linkage to three polymorphic markers within p53 enabled direct involvement of p53 to be excluded. This is the first example of a LFS family in whom exclusion of p53 has been possible. Four affected members of the family with sarcoma or premenopausal breast cancer showed increased expression of p53 protein in their normal tissues as detected by immunohistochemistry. It therefore appears that the LFS phenotype has been conferred by an aberrant gene, showing a dominant pattern of inheritance, which may be acting to compromise normal p53 function rather than by a mutation in p53 itself. In order to try to determine the chromosomal location of this putative gene, we have carried out studies of linkage to candidate loci. By these means we have excluded involvement of Rb1 and BRCA1 on chromosomes 13q and 17q respectively. The MDM2 oncogene on chromosome 12q was considered to be the prime candidate as MDM2 is amplified in sarcomas and the MDM2 product binds to p53. Furthermore, p53 mutation and amplification of MDM2 have been shown to be mutually exclusive events in tumour development. Linkage analysis to two polymorphic markers within MDM2 yielded a three-point LOD score of -5.4 at a recombination fraction theta equal to zero. Therefore MDM2 could be excluded. It is possible that the gene which is responsible for cancer susceptibility in this family, possibly via interaction with p53, will be important in the histogenesis of breast cancer in general. We are now carrying out further studies to locate and identify this gene.
    • A linkage study in seven breast cancer families.

      Teare, M Dawn; Santibanez-Koref, Mauro F; Wallace, S A; White, Gavin R M; Evans, D Gareth R; Burnell, Liza D; Harris, Martin; Howell, Anthony; Birch, Jillian M; Cancer Research Campaign Pediatric and Familial Cancer Research Group, Christie Hospital, Manchester, England. (1993-04)
      Seven breast cancer families are examined for evidence of linkage to a site in the region of 17q12-q21, by using five markers. The families constitute a subset of a larger series of familial breast cancer; the seven families were selected because constitutional DNA was available on informative members, either from clinical samples or extracted from paraffin blocks. Two-point lod scores are reported. The maximum lod score, 0.8824, is obtained with marker NM23 at theta = 0. This is clearly not significant in itself; however, when taken in context with evidence from existing reports, it provides support for linkage to this region.
    • Linked parallel synthesis and MTT bioassay screening of substituted chalcones.

      Lawrence, Nicholas J; Rennison, David; McGown, Alan T; Ducki, Sylvie W; Gul, Lubna A; Hadfield, John A; Khan, Nader; Department of Chemistry, University of Manchester Institute of Science and Technology, P.O. Box 88, Manchester, M60 1QD, UK. Lawrencenj1@cardiff.ac.uk (2001)
      A 644-membered library of chalcones was prepared by parallel synthesis using the Claisen-Schmidt base-catalyzed aldol condensation of substituted acetophenones and benzaldehydes. The cytotoxicity of these chalcones was conveniently determined upon the crude products directly in 96-well microtiter test plates by the conventional MTT assay. This method revealed seven chalcones of IC(50) less than 1 microM of which 4'-hydroxy-2,4,6,3'-tetramethoxychalcone (5a) was the most active [IC(50) (K562), 30 nM]; it causes cell cycle arrest at the G(2)/M point and binds to tubulin at the colchicine binding site.
    • The lipid fluidity of rat liver membrane subfractions.

      Whetton, Anthony D; Houslay, M D; Dodd, Nicholas J F; Evans, W H; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Withington, Manchester M20 9BX (1983-09-15)
      1. The lipid fluidity of three major rat liver plasma-membrane subfractions, as well as Golgi apparatus and endocytic fractions, was assessed with a fatty acid spin probe by using e.s.r. techniques. 2. The sinusoidal (blood-facing) plasma-membrane subfraction was the most fluid of the three plasma-membrane regions. Fractions originating from the bile-canalicular and contiguous (lateral) regions were most rigid. Endocytic fractions isolated (endosomes and diacytosomes) were of a similar fluidity to fractions originating from the sinusoidal plasma-membrane region. By far the most fluid fractions examined were derived from the Golgi-apparatus complex. 3. The three plasma-membrane subfractions each showed a different response to the bilayer-fluidizing effect of benzyl alcohol. 4. Arrhenius-type plots of the order parameter S and outer hyperfine splitting, 2T( parallel), identified lipid-phase separations in the plasma-membrane subfractions.
    • Liquid biopsy for advanced non-small cell lung cancer: a consensus statement from the international association for the study of lung cancer (IASLC)

      Rolfo, C.; Mack, P.; Scagliotti, G. V.; Aggarwal, C.; Arcila, M. E.; Barlesi, F.; Bivona, T.; Diehn, M.; Dive, Caroline; Dziadziuszko, R.; et al. (2021)
    • A liquid scintillation counting method that allows recovery of compounds in high yield for further analysis.

      Saffhill, Roy; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester, M20 9BX. United Kingdon (1981-06)
    • The live vector approach-viruses.

      Mackett, Mike (1991-03)
      Recombinant DNA technology has made it possible to express foreign genes in viruses and bacteria. This has led to the idea of engineering live attenuated vaccines to express protective antigens from foreign pathogens. Vaccinia virus, the smallpox vaccine, has been used to ploneer this approach. Several hundred recombinants have been recorded in the literature and many of them have been shown to protect animals against challenge with the appropriate pathogen. Although mixed results have been obtained with experiments in humans and primates, and there is some concern over complications associated with vaccination, recent advances in our under-standing of the basic biology of vaccinia virus should allow these difficulties to be overcome. Whatever the final outcome of proposals to use vaccinia recombinants in humans, vaccinia has proved to be a valuable general purpose expression system and is particularly useful for studying cellular immunity. The success of field trials with a vaccinia recombinant expressing the rabies virus glycoprotein may lead to widespread use of vaccinia recombinants as animal vaccines. Therefore it seems likely that vaccinia recombinants will continue to play a useful part in the development of new vaccines for infectious disease.
    • Liver heparan sulfate structure. A novel molecular design.

      Lyon, Malcolm; Deakin, Jon A; Gallagher, John T; Cancer Research Campaign, Christie Hospital, Manchester, United Kingdom. (1994-04-15)
      The structure of rat liver heparan sulfate (HS) has been investigated using a combination of (a) chain scission with specific reagents, (b) disaccharide compositional analysis, and (c) end-referenced sequence analysis of the proximal, protein-linked region of the chain. This study reveals that the liver synthesizes a highly sulfated HS species (1.34 sulfates/disaccharide), particularly high in N-sulfation (60%) and 2-O-sulfate content (36%). Approximately half of the latter is found in trisulfated disaccharides, i.e. IdceA(2-OSO3) alpha 1-4GlcNSO3 (6-OSO3). End-referencing methodology established the existence of an extended, unmodified heparan (GlcUA beta 1-4GlcNAc) sequence, 8-11 disaccharides in length, attached to the linkage tetrasaccharide, similar to that found in a number of other HS species. Directly following this is a mixed HexUA1-4GlcNR(6-OSO3) (where GlcNR represents alpha-D-glucosamine with an unspecified N-substituent)-containing sequence of variable length, culminating in the appearance of the first IdceA(2-OSO3) residue approximately 20 disaccharides from the linkage region, i.e. approximately 40% along the length of the chain. The distal 60% of the polysaccharide is highly sulfated (approximately 2 sulfates/disaccharide) and mainly comprises three heparin-like domains, highly enriched in IdceA(2-OSO3) residues. Overall, liver HS qualifies as an extreme member of the HS family, with a considerable proportion of heparin-like structure asymmetrically concentrated to the distal part of the chain.
    • The local anaesthetic and bilayer fluidising agent, benzyl alcohol decreases the thermostability of the integral membrane protein adenylate cyclase

      Needham, L; Whetton, Anthony D; Houslay, M; Department of Biochemistry, UMIST, PO BOX 88, Manchester M60 1QD. (1982-04)
    • Localisation of granulocyte macrophage colony-stimulating factor in human long-term bone marrow cultures. Biological and immunocytochemical characterisation.

      De Wynter, Erika A; Allen, Terence D; Coutinho, Lucia H; Flavell, D; Flavell, S U; Dexter, T Michael; Cancer Research Campaign Department of Experimental Haematology, Paterson Institute for Cancer Research, Manchester, UK. (1993-11)
      The distribution of granulocyte macrophage colony-stimulating factor (GM-CSF) in human long-term bone marrow cultures (HLTBMC) was examined using two monoclonal antibodies raised using purified recombinant GM-CSF and a third commercially available GM-CSF antibody. The antibodies were able to bind to purified recombinant GM-CSF and showed inhibition of GM-CFC colonies in the presence of both recombinant and native protein. All antibodies displayed similar patterns of distribution in both permeabilised and non-permeabilised stromal cell preparations. Fibroblasts were labelled at their periphery in early cultures and both endothelial cells and fibroblasts showed cytoplasmic labelling with anti-GM-CSF. The fact that GM-CSF appears to be sequestered by cells of the bone marrow stroma raises the possibility that it is synthesized by these cells and may regulate activity of the progenitor cells in the haemopoietic foci. In contrast, early progenitor cells within the foci did not stain with any of the anti-GM-CSF antibodies. Adipocytes, which differentiate from fibroblasts in these cultures, showed a diffuse staining pattern. Two types of macrophage staining were observed in the non-permeabilised cells; those exhibiting only autofluorescence and those that bound the antibody. Intracellular staining was apparent in a small sub-population. Generally, the staining persisted up to eight weeks of culture and thereafter declined, becoming virtually undetectable after 12 weeks. This correlates with the pattern of GM-CFC production in long-term bone marrow cultures.
    • Locally advanced lung cancer radiotherapy in deep inspiration breath hold: dosimetric benefits from a prospective trial

      Josipovic, M; Aznar, Marianne Camille; Rydhog, J; Thomsen, J; Damkjaer, S; Nygard, L; Pohl, M; Langer, S; Specht, L; Persson, G; et al. (2018)