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Biomedical subjects

C W Evans

Publications and source records attributed to C W Evans.

At least 19 recordsLinked to original sources

A comparison of the technical sustainability of in situ stabilisation/solidification with disposal to landfill.

Sustainability is becoming a very important issue in contaminated land remediation and should form one of the factors used in future selection of treatment technologies. In situ stabilisation/solidification (S/S) is a remediation technique that is increasingly being applied to the treatment of contaminated sites because of numerous advantages over other remediation techniques. This paper assesses and compares aspects of the technical sustainability of in situ S/S with landfilling. Criteria previously established for the assessment of the technical sustainability of the remediation of contaminated land are employed. The comparison is presented in the form of a case study based on a real remediation project in the UK. The analysis indicated that landfilling had a larger impact than S/S in the majority of areas investigated, such as waste production (1000 kg waste/t soil remediated for landfilling compared to none for S/S), transportation (12.9 km/t for landfilling, 0.4 km/t for S/S) and use of raw materials (1005.5 kg/t for landfilling, 88.9 kg/t for S/S), although S/S had high greenhouse gas emissions (12.6 kg/t for landfilling, 40.9 kg/t for S/S). In addition, a multi-criteria/cost-effectiveness analysis gave cost effectiveness scores of -34.2 to S/S and -138.1 to landfill (where more positive is better).

Environmental Restoration and Remediation↗

Analysis of the circadian clock gene period in the sheep blow fly Lucilia cuprina.

We have isolated a homologue of the period (per) gene from the Australian sheep blow fly, Lucilia cuprina, as part of a comparative approach to the analysis of dipteran circadian systems. Sequence analysis of the 4 kb per cDNA revealed the conservation of three functional domains, namely the PAS dimerization motif, and the nuclear and cytoplasmic localization domains. A fourth domain, the threonine-glycine (TG) repeat region, is also conserved in L. cuprina per but has been severely truncated. No length variation was found in the TG repeat of L. cuprina or L. sericata collected from several different latitudinal zones. Expression analysis indicated a diel oscillation in per mRNA in LD 12:12 with a period of 24 h and a peak at Zt 12. PER-immunoreactive protein oscillations were also demonstrated, with peak immunoreactivity lagging approximately 3 h behind peak mRNA levels. These results show the existence of a Drosophila-like circadian system in a calliphorid fly. They also provide evidence for the conservation of per function across the Diptera, and confirm the relevance of the Drosophila system as a model for fly circadian rhythms.

Amino Acid Sequence↗

Antigen recognition and activation of ovine gamma delta T cells.

We have investigated several aspects of gamma delta T cells in sheep. gamma delta T cells of sheep express a unique transmembrane protein termed T19 but lack the expression of particular cell-surface molecules such as CD2, CD4 and CD8 which are typically associated with alpha beta T cells. The majority of gamma delta T cells isolated from animals of all ages examined lacked the expression of CD45RA. A faster rate of activation by gamma delta T cells compared to either CD4 or CD8 T cells was seen in the time-course of IL-2 receptor alpha chain (CD25) cell-surface expression. All gamma delta T cells expressed the CD25 protein within 8 hr of activation whereas the majority of CD4 or CD8 T cells did not express CD25 until 24 hr post-concanavalin A (Con A) stimulation. This difference in the rate of expression of activation molecules was not restricted to CD25, as a similar trend was seen with cell-surface expression of major histocompatibility complex (MHC) class II molecules. We have used the distinct phenotypic profile of ovine gamma delta T cells to purify these cells by positive selection via the T19 molecule to assess their in vitro proliferative response to various antigens. Routinely, cell populations comprising more than 93% gamma delta T cells with yields of approximately 55% were obtained. Purified gamma delta T cells were capable of responding to Mycobacterium tuberculosis antigen in a primary and secondary in vitro proliferation assay and to ovalbumin in a secondary response. Ovine gamma delta T cells showed little, if any, proliferative response to allogeneic stimulator cells.

Animals↗

Different rates of interleukin 2 receptor expression by ovine gamma/delta and alpha/beta T cells.

We have used the monoclonal antibodies IL-A111, CACT116A, to investigate the expression of interleukin 2 receptors (IL-2R) by ovine peripheral blood T cells. Monoclonal antibodies (MAbs) IL-A111 and CACT116A reacted with concanavalin A (Con A) activated ovine lymphocytes and both mAbs did so with similar FACS (fluorescence activated cell sorting) profiles and fluorescence intensity. Following activation by Con A all three major ovine T cell subsets, namely CD4+, CD8+ and gamma/delta T cells, express the IL-2R. All gamma/delta T cells express IL-2R within 12 h following activation by Con A in vitro whereas alpha/beta T cells do not express the IL-2R until 24 h after the start of culture with the mitogen. In freshly isolated peripheral blood a small percentage of lymphocytes, which are almost all CD4+, express the IL-2R.

Amino Acid Sequence↗

Expression of T19 (WC1) molecules by ovine lymphocytes.

Workshop clusters WC1 and WC2 monoclonal antibodies (mAbs) were tested for their reactivity with ovine peripheral blood mononuclear cells (PBMC). The populations of ovine lymphocytes defined by these mAbs were found to be part of the population of T19+ (WC1+) cells, and the gamma/delta T cell receptor (TCR) (WC2) expressing T cells. The expression of cell surface WC1 antigens following mitogen stimulation of ovine PBMC was studied. Whilst the size of the WC1 populations increased during culture with concanavalin A, the changes in the ratio of WC1:gamma/delta TCR percentages observed suggested either a loss of WC1 molecules or a selective expansion of WC1- cells.

Animals↗

Gap junction formation during development of the mouse lens.

We have identified a 60 kDa membrane protein (MP60) as a component of the mouse cortical lens fiber gap junction and a monoclonal antibody recognizing this protein has been used to establish the temporal and spatial patterns of gap junction formation during development of the mouse lens. The initial expression of MP60 during embryonic development of the mouse lens correlates with primary fiber elongation and is first seen on the luminal aspect of the extending cells. About 2 days after birth, the relatively large, antibody-positive macular structures characteristic of late embryonic fiber cells begin to disperse into progressively smaller structures within a centrally located region of the lens. This change in the staining pattern with antibody directed against MP60 is consistent with the dispersion of gap junction plaques as confirmed by freeze fracture analysis. Around 5 days after birth, the 60 kDa gap junctional protein in this central region of the lens undergoes a modification resulting in the alteration to the epitope for the monoclonal antibody and a consequent loss of immunorecognition. Our results suggest that gap junctions in the central region of the developing mouse lens undergo sequential changes in immunoreactivity which may reflect potentially distinct functional phases of intercellular communication.

Age Factors↗

Cell adhesion and metastasis.

There is no single phenotypic trait whose exclusive expression correlates universally with metastatic behaviour. Nevertheless, several lines of evidence point towards adhesive phenomena as mediating some crucial steps in the malignant spread of particular tumour types. These steps include detachment from the primary, invasion of the ECM including include detachment from the primary, invasion of the ECM including structures such as the basement membrane and organ capsules, and lodgement in the vessels of remote organs. Furthermore, there is evidence that adhesive phenomena can contribute at least in part to the selective patterns of spread shown by a variety of tumour cell types. The molecular dissection of the adhesive events involved in these processes has only just begun. It is clear already, however, that several different determinants are likely to be involved and the clinical application of this knowledge is unlikely to be immediate.

Amino Acid Sequence↗

A genetic basis for metastasis.

The progression of a normal cell to one that is malignant is characterized by at least four progressive but potentially separable behavioural patterns identifying the immortal, tumorigenic, invasive and metastatic phenotypes. A multitude of steps appear to be involved in both transformation from normality and progression to malignancy as characterized by the acquisition of metastatic behaviour. Consequently, it seems unlikely that a single gene can directly manifest expression of the metastatic phenotype in normal cells unless it can induced pleiotropic effects. Indeed, a single trait uniquely characterizing the metastatic phenotype has never been identified. The possibility of a single gene suppressing the metastatic phenotype seems much greater. One possible candidate for such a gene is nm23, the expression of which correlates with reduced metastatic potential in several tumours including breast cancer in humans. Although the numbers involved are still small, the correlation of nm23 expression with breast cancer outcome offers potential in using this system as a prognostic aid in clinical diagnosis of this disease. Its possible role as an indicator of metastatic potential in other human tumours remains to be evaluated, although current evidence suggests that it is unlikely to be of use in colon cancer. Further significant progress requires molecular dissection of the mode of action of its product.

Humans↗

Molecular portrait of lens gap junction protein MP70.

A 70-kDa membrane protein (MP70) is a component of the lens fiber gap junctions. Its membrane topology and its N-terminal sequence are similar to those of the connexin family of proteins. Some features of MP70 containing fiber gap junctions are, however, distinct from gap junctions in other mammalian tissues: (i) Lens connexons form crystalline arrays only after cleavage of junctional proteins in vitro. These hexagonal arrays have a periodicity of 13.6 nm which is significantly larger than the 8- 9-nm spacing of liver and heart gap junctions. (ii) Lens fiber gap junctions dissociate in low concentrations of nonionic detergent and this provides an avenue to purify MP70 directly from a membrane mixture. Isolated MP70 in the form of 17 S structures has an appearance consistent with connexon pairs. (iii) The C-terminal half of MP70 is cleaved in situ by a lens endogenous calcium-dependent protease. The processed from MP38 remains in the membrane and is abundant in the central region of the lens. A testable hypothesis for MP70 function is presented.

Connexins↗

Plasma catecholamines and their physiologic thresholds during the first ten days of life in sheep.

We studied serial plasma catecholamine levels in healthy newborn sheep over the first ten days of life. The results show that plasma norepinephrine values in newborn sheep are 3-4 fold higher, and plasma epinephrine values are two-fold higher than values in term fetal sheep. These elevations are sustained over the first 10 days of life. Cardiovascular (heart rate and blood pressure) and metabolic parameters (glucose and free fatty acids) are also significantly elevated above fetal levels. We performed graded catecholamine infusions in newborn animals and adult ewes to determine the minimum plasma catecholamine concentrations necessary for discernible physiologic effects. In response to step-wise increases in epinephrine or norepinephrine infusion rates, there were immediate increases in blood pressure and other physiologic responses. This pattern was seen in both newborn and adult animals, and differed from previous observations in fetal sheep where log-linear, dose response curves characteristic of a threshold response were seen. These results suggest that during the first two weeks of life plasma catecholamine levels are elevated above the threshold value for physiologic responses. These sustained elevations in circulating catecholamines are important in the maintenance of physiologic homeostasis.

Animals↗

Neutrophil chemotactic behaviour in patients with early-onset forms of periodontitis (I). Leading front analysis in Boyden chambers.

Despite some reports to the contrary, it is generally assumed that early-onset forms of periodontal disease (including both juvenile and rapidly progressive periodontitis) are associated with a defect in neutrophil (PMN) chemotactic behaviour. Using the Boyden chamber technique and N-formyl-methionyl-leucylphenylalanine (FMLP) to assess locomotion by the leading front method, we have failed to show any evidence for such depressed PMN locomotion. Indeed, when PMN chemotaxis and chemokinesis are considered in response to a range of chemoattractant doses our results indicate significant enhancement of all but random locomotion. When taken together with other studies, our results suggest that PMNs from patients with early-onset periodontitis may show abnormal locomotory behaviour which can either be enhanced or reduced in nature. The extent to which these results reflect in vitro methodology in uncertain and, furthermore, their in vivo significance is unclear.

Adolescent↗

Neutrophil chemotactic behaviour in patients with early-onset forms of periodontitis (II). Assessment using the under agarose technique.

The locomotory behaviour of peripheral blood neutrophils (PMNs) from patients with juvenile (JP) and rapidly progressive (RPP) forms of early-onset periodontal disease was studied using the under agarose technique and n-formyl-methionyl-leucyl-phenylalanine (FMLP) as the chemotractant. PMNs from experimental patients showed normal random, chemotactic and chemokinetic locomotory behaviour when compared with control subjects. Further investigation of single-cell movements using time-lapse video analysis also failed to show any significant differences in locomotory behaviour between the PMNs of experimental and control individuals. We conclude that differences in technique may account for much of the variation which exists in the literature with respect to PMN locomotion in periodontal disease. In the final analysis, it is difficult to dispute direct observation of moving cells, and using this approach, we have been unable to confirm the presence of any PMN locomotory defect in our series of patients with early-onset periodontal disease.

Adolescent↗

Depressed helper-to-suppressor T-cell ratios in early-onset forms of periodontal disease.

Helper (CD4+)-to-suppressor (CD8+) T-cell ratios were determined by flow cytometry for 12 patients with early-onset forms of periodontal disease and their appropriate controls. When considered as a group, patients with early-onset periodontal disease showed a markedly depressed CD4+/CD8+ ratio relative to controls (means 0.92 and 1.50 respectively). This difference was significant at the 0.0015% level as determined by the Mann-Whitney test. Separate analysis of patients with either the juvenile or rapidly progressive types of early-onset periodontal disease showed both to have reduced CD4+/CD8+ ratios (means 0.91 and 0.92) relative to their controls (means 1.57 and 1.45) which were both statistically significant (p less than 0.05).

Adolescent↗