I am a senior academic and Executive Dean, with a 45year history and professional background in service pathology and medical research. A specialist in the field of oxidative damage to biomolecules and its significance for disease process.
Monocytes play a central role in inflammatory responses through systemic antigen presentation and... more Monocytes play a central role in inflammatory responses through systemic antigen presentation and cytokine secretion. Regulation of monocyte adhesion molecule and inflammatory gene expression is via redox sensitive transcription factors. Therefore we have investigated the hypothesis that dietary antioxidant supplementation with vitamins C (250mg/d) or E (400iU/d) for six weeks can modulate monocyte ICAM-1 expression in healthy male subjects with low plasma vitamin C at baseline. In a randomised, double-blind, crossover study, ICAM-1 mRNA and protein was analysed using quantitative RTPCR with ELISA measurement of PCR products and by flow cytometry and ELISA respectively. Monocyte numbers were unaltered by supplementation. Subjects with low plasma vitamin C (<50uM) prior to supplementation expressed higher levels of monocyte ICAM-1 mRNA, and showed a significant (50%) reduction in ICAM-1 mRNA expression after 6 weeks of 250mg/d vitamin C supplementation compared to subjects with normal plasma vitamin C. This was paralleled by a reduction in plasma sICAM-1. Vitamin E supplementation had no effect on ICAM-1 expression. For the first time, these results show that dietary vitamin C can modulate monocyte ICAM-1 gene expression in vivo, where regulation of gene expression represents a novel mechanism for benefit from dietary antioxidants.
The repair of oxidatively damaged DNA is integral to the maintenance of genomic stability, and he... more The repair of oxidatively damaged DNA is integral to the maintenance of genomic stability, and hence prevention of a wide variety of pathological conditions, such as aging, cancer and cardiovascular disease. The ability to non-invasively assess DNA repair may provide information regarding repair pathways, variability in repair capacity, and susceptibility to disease. The development of assays to measure urinary DNA lesions offered this potential, although it rapidly became clear that possible contribution from diet and cell turnover may influence urinary lesion levels. Whilst early studies attempted to address these issues, up until now, much of the data appears conflicting. However, recent work from our laboratories, in which human volunteers were fed highly oxidatively modified 15N-labelled DNA demonstrates that diet does not appear to contribute to urinary levels of 8-hydroxyguanine and 7,8-dihydro-8-oxo-2'-deoxyguanosine. Furthermore, we propose that a number of literature r...
Page 1. 222 ANALYTICAL PROCEEDINGS, AUGUST 1990, VOL 27 Protein Fluorescence: its Generation and ... more Page 1. 222 ANALYTICAL PROCEEDINGS, AUGUST 1990, VOL 27 Protein Fluorescence: its Generation and Measurement Helen R. Griffiths and Joseph Lunec Wolfson Research Laboratories, Queen Elizabeth Medical Centre, Birmingham B 75 2TH ...
This review is concerned with the tumour necrosis factor receptor and ligand superfamilies, with ... more This review is concerned with the tumour necrosis factor receptor and ligand superfamilies, with particular reference to their roles in the immunopathogenesis of Systemic Lupus Erythematosus (SLE). The tumour necrosis factor receptor and ligand superfamilies are well characterized as the molecular controllers of the immune system, acting as &#39;judges&#39;, &#39;juries&#39;, and, where necessary, &#39;executioners&#39; to determine the fate of immune cells during development, proliferation and differentiation. However, these molecules exert extreme immunopathological effects when unregulated, or dysfunctional. The importance of these molecules in the pathogenesis of autoimmunity is now apparent, and has been considered in detail. Finally, specific consideration has been given to their clinical significance and potential diagnostic and therapeutic applications.
References I11 F. HmATA, E. SCH[FFMANN, K. VENgATASUBRAMANIAN, D. SALOMON and J. AXELROD. A phosp... more References I11 F. HmATA, E. SCH[FFMANN, K. VENgATASUBRAMANIAN, D. SALOMON and J. AXELROD. A phospholipase A 2 inhibitory protein in rabbit neutrophils induced by glucocorticoids. Proc. natn. Acad. Sci. USA 77, 2533-2536 (1980). I21 G.J. BLACKWELL, R. CARNUCCIO, M. DI ROSA, R.J. FLOWER, C.S.J. LANGI-IAM, L. PARENTE, P. PERSICO, N.C. RUSSELL SMITH and D. STONE, Glucocorticoids induce the formation and release of anti-inflammator.i, and anti-phospholipase proteins into the peritoneal ca~,ity of the rat, Br. J. Pharmac. 76, 185-194 (1982). 131 F. RUSSO-MARIE, M. PAING and D. DUVAL, Im, olvement of glucocorticoid receptors in steroidI41
“There can be no more important goal in a clinical laboratory than that of ensuring that the resu... more “There can be no more important goal in a clinical laboratory than that of ensuring that the results produced have the precision and accuracy necessary to aid diagnosis and treatment.”T.P. Whitehead 1976
Global gene expression profiles of livers from mice, fed diets differing in alpha-tocopherol cont... more Global gene expression profiles of livers from mice, fed diets differing in alpha-tocopherol content, were compared using DNA microarray technology. Three hundred and eighty nine genes were found to significantly differ in their expression level by a factor of 2 or higher between the high and the low alpha-tocopherol group. Functional clustering using the EASE software identified 121 genes involved in transport processes. Twenty-one thereof were involved in (synaptic) vesicular trafficking. Up-regulation of syntaxin 1C (Stx1c), vesicle-associated membrane protein 1 (Vamp1), N-ethylmaleimide-sensitive factor (Nsf) and syntaxin binding protein 1 (Stxbp1, Munc18-1) was verified by real time PCR. At a functional level, alpha-tocopherol increased the secretory response in RBL and PC12 cells. Although here detected in liver, the alpha-tocopherol-responsive pathways are also relevant to neurotransmission. A role of alpha-tocopherol in the vesicular transport might not only affect its own absorption and transport but also explain the neural dysfunctions observed in severe alpha-tocopherol deficiency.
Six patients undergoing vascular reconstructive surgery were examined for evidence of oxygen-deri... more Six patients undergoing vascular reconstructive surgery were examined for evidence of oxygen-derived free radical (ORF) damage to the protein, immunoglobulin G (IgG). OFR damage was determined as an increase in the fluorescence (ex 360 nm em 454 nm) to ultraviolet absorption (280 nm) ratio of IgG, representing N-Formyl kynurenine and other as yet unidentified fluorophores. The IgG ratio was found to increase slightly during ischaemia and to undergo marked elevation upon reperfusion (275 +/- 405% baseline value at 40 min post-clamp; mean +/- sd). A high ratio was maintained post-reperfusion, even after 60 min reperfusion. Determination of thromboxane B2, (TXB2), leukotriene B4, (LTB4) and 6-keto prostaglandin F1 alpha, (PGF1a), revealed a decrease in their concentrations during ischaemia and a transient, marked increase on reperfusion. Only TXB2 concentrations were found to correlate with the IgG ratio (negative correlation, p &lt; 0.05). No correlation was observed between von Willebrand antigen factor, a marker of endothelial cell damage and fluorescent IgG ratio. However, levels of the factor increased slightly during ischaemia and more sharply upon reperfusion. These preliminary results therefore suggest that a more likely source of the OFRs responsible for IgG damage is endothelial cell xanthine oxidase, rather than cyclo-oxygenase or lipoxygenase.
In this research we developed a real-time SYBR green assay to detect both Mycoplasma gallisepticu... more In this research we developed a real-time SYBR green assay to detect both Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) in a single reaction. A total of 30,000 samples from broiler breeder flocks were screened using traditional serology (plate agglutination, enzyme-linked immunosorbent assay, hemagglutination inhibition) and polymerase chain reaction (PCR; traditional and real-time). It was determined that the real-time SYBR green PCR assay developed in this research was more rapid than all three methods tested and more sensitive and specific than culturing or serology. The SYBR green assay was optimized and could detect as few as 30 template copies of DNA per sample. In addition, the SYBR green assay was less expensive than traditional culturing and serology. MG and MS are infectious bacteria that can rapidly spread and infect commercial chicken flocks. These diseases can cause a significant loss to the poultry industry and especially to broiler breeders because infected flocks are destroyed under the National Poultry Improvement Plan MG and MS clean programs. The real-time SYBR green assay developed in this research has the potential to reduce the time it takes to reach a correct diagnosis and to arrest outbreaks of MG and MS.
Monocytes play a central role in inflammatory responses through systemic antigen presentation and... more Monocytes play a central role in inflammatory responses through systemic antigen presentation and cytokine secretion. Regulation of monocyte adhesion molecule and inflammatory gene expression is via redox sensitive transcription factors. Therefore we have investigated the hypothesis that dietary antioxidant supplementation with vitamins C (250mg/d) or E (400iU/d) for six weeks can modulate monocyte ICAM-1 expression in healthy male subjects with low plasma vitamin C at baseline. In a randomised, double-blind, crossover study, ICAM-1 mRNA and protein was analysed using quantitative RTPCR with ELISA measurement of PCR products and by flow cytometry and ELISA respectively. Monocyte numbers were unaltered by supplementation. Subjects with low plasma vitamin C (<50uM) prior to supplementation expressed higher levels of monocyte ICAM-1 mRNA, and showed a significant (50%) reduction in ICAM-1 mRNA expression after 6 weeks of 250mg/d vitamin C supplementation compared to subjects with normal plasma vitamin C. This was paralleled by a reduction in plasma sICAM-1. Vitamin E supplementation had no effect on ICAM-1 expression. For the first time, these results show that dietary vitamin C can modulate monocyte ICAM-1 gene expression in vivo, where regulation of gene expression represents a novel mechanism for benefit from dietary antioxidants.
The repair of oxidatively damaged DNA is integral to the maintenance of genomic stability, and he... more The repair of oxidatively damaged DNA is integral to the maintenance of genomic stability, and hence prevention of a wide variety of pathological conditions, such as aging, cancer and cardiovascular disease. The ability to non-invasively assess DNA repair may provide information regarding repair pathways, variability in repair capacity, and susceptibility to disease. The development of assays to measure urinary DNA lesions offered this potential, although it rapidly became clear that possible contribution from diet and cell turnover may influence urinary lesion levels. Whilst early studies attempted to address these issues, up until now, much of the data appears conflicting. However, recent work from our laboratories, in which human volunteers were fed highly oxidatively modified 15N-labelled DNA demonstrates that diet does not appear to contribute to urinary levels of 8-hydroxyguanine and 7,8-dihydro-8-oxo-2'-deoxyguanosine. Furthermore, we propose that a number of literature r...
Page 1. 222 ANALYTICAL PROCEEDINGS, AUGUST 1990, VOL 27 Protein Fluorescence: its Generation and ... more Page 1. 222 ANALYTICAL PROCEEDINGS, AUGUST 1990, VOL 27 Protein Fluorescence: its Generation and Measurement Helen R. Griffiths and Joseph Lunec Wolfson Research Laboratories, Queen Elizabeth Medical Centre, Birmingham B 75 2TH ...
This review is concerned with the tumour necrosis factor receptor and ligand superfamilies, with ... more This review is concerned with the tumour necrosis factor receptor and ligand superfamilies, with particular reference to their roles in the immunopathogenesis of Systemic Lupus Erythematosus (SLE). The tumour necrosis factor receptor and ligand superfamilies are well characterized as the molecular controllers of the immune system, acting as &#39;judges&#39;, &#39;juries&#39;, and, where necessary, &#39;executioners&#39; to determine the fate of immune cells during development, proliferation and differentiation. However, these molecules exert extreme immunopathological effects when unregulated, or dysfunctional. The importance of these molecules in the pathogenesis of autoimmunity is now apparent, and has been considered in detail. Finally, specific consideration has been given to their clinical significance and potential diagnostic and therapeutic applications.
References I11 F. HmATA, E. SCH[FFMANN, K. VENgATASUBRAMANIAN, D. SALOMON and J. AXELROD. A phosp... more References I11 F. HmATA, E. SCH[FFMANN, K. VENgATASUBRAMANIAN, D. SALOMON and J. AXELROD. A phospholipase A 2 inhibitory protein in rabbit neutrophils induced by glucocorticoids. Proc. natn. Acad. Sci. USA 77, 2533-2536 (1980). I21 G.J. BLACKWELL, R. CARNUCCIO, M. DI ROSA, R.J. FLOWER, C.S.J. LANGI-IAM, L. PARENTE, P. PERSICO, N.C. RUSSELL SMITH and D. STONE, Glucocorticoids induce the formation and release of anti-inflammator.i, and anti-phospholipase proteins into the peritoneal ca~,ity of the rat, Br. J. Pharmac. 76, 185-194 (1982). 131 F. RUSSO-MARIE, M. PAING and D. DUVAL, Im, olvement of glucocorticoid receptors in steroidI41
“There can be no more important goal in a clinical laboratory than that of ensuring that the resu... more “There can be no more important goal in a clinical laboratory than that of ensuring that the results produced have the precision and accuracy necessary to aid diagnosis and treatment.”T.P. Whitehead 1976
Global gene expression profiles of livers from mice, fed diets differing in alpha-tocopherol cont... more Global gene expression profiles of livers from mice, fed diets differing in alpha-tocopherol content, were compared using DNA microarray technology. Three hundred and eighty nine genes were found to significantly differ in their expression level by a factor of 2 or higher between the high and the low alpha-tocopherol group. Functional clustering using the EASE software identified 121 genes involved in transport processes. Twenty-one thereof were involved in (synaptic) vesicular trafficking. Up-regulation of syntaxin 1C (Stx1c), vesicle-associated membrane protein 1 (Vamp1), N-ethylmaleimide-sensitive factor (Nsf) and syntaxin binding protein 1 (Stxbp1, Munc18-1) was verified by real time PCR. At a functional level, alpha-tocopherol increased the secretory response in RBL and PC12 cells. Although here detected in liver, the alpha-tocopherol-responsive pathways are also relevant to neurotransmission. A role of alpha-tocopherol in the vesicular transport might not only affect its own absorption and transport but also explain the neural dysfunctions observed in severe alpha-tocopherol deficiency.
Six patients undergoing vascular reconstructive surgery were examined for evidence of oxygen-deri... more Six patients undergoing vascular reconstructive surgery were examined for evidence of oxygen-derived free radical (ORF) damage to the protein, immunoglobulin G (IgG). OFR damage was determined as an increase in the fluorescence (ex 360 nm em 454 nm) to ultraviolet absorption (280 nm) ratio of IgG, representing N-Formyl kynurenine and other as yet unidentified fluorophores. The IgG ratio was found to increase slightly during ischaemia and to undergo marked elevation upon reperfusion (275 +/- 405% baseline value at 40 min post-clamp; mean +/- sd). A high ratio was maintained post-reperfusion, even after 60 min reperfusion. Determination of thromboxane B2, (TXB2), leukotriene B4, (LTB4) and 6-keto prostaglandin F1 alpha, (PGF1a), revealed a decrease in their concentrations during ischaemia and a transient, marked increase on reperfusion. Only TXB2 concentrations were found to correlate with the IgG ratio (negative correlation, p &lt; 0.05). No correlation was observed between von Willebrand antigen factor, a marker of endothelial cell damage and fluorescent IgG ratio. However, levels of the factor increased slightly during ischaemia and more sharply upon reperfusion. These preliminary results therefore suggest that a more likely source of the OFRs responsible for IgG damage is endothelial cell xanthine oxidase, rather than cyclo-oxygenase or lipoxygenase.
In this research we developed a real-time SYBR green assay to detect both Mycoplasma gallisepticu... more In this research we developed a real-time SYBR green assay to detect both Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) in a single reaction. A total of 30,000 samples from broiler breeder flocks were screened using traditional serology (plate agglutination, enzyme-linked immunosorbent assay, hemagglutination inhibition) and polymerase chain reaction (PCR; traditional and real-time). It was determined that the real-time SYBR green PCR assay developed in this research was more rapid than all three methods tested and more sensitive and specific than culturing or serology. The SYBR green assay was optimized and could detect as few as 30 template copies of DNA per sample. In addition, the SYBR green assay was less expensive than traditional culturing and serology. MG and MS are infectious bacteria that can rapidly spread and infect commercial chicken flocks. These diseases can cause a significant loss to the poultry industry and especially to broiler breeders because infected flocks are destroyed under the National Poultry Improvement Plan MG and MS clean programs. The real-time SYBR green assay developed in this research has the potential to reduce the time it takes to reach a correct diagnosis and to arrest outbreaks of MG and MS.
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Papers by Joseph Lunec