Jane MacKenzie

Detection of Epstein-Barr virus (EBV) genomes in the serum of patients with EBV-associated Hodgkin's disease

DNA from malignant cells is present in the serum/plasma of cancer patients and DNA from this source is amenable to analysis by polymerase chain reaction (PCR). In the present study, we evaluated whether Epstein‐Barr virus (EBV) DNA is present in the serum of patients with EBV‐associated Hodgkins disease (HD). Using conventional PCR, EBV DNA was detected in serum from 30/33 patients with EBV‐associated HD but in only 6/26 patients with non‐EBV‐associated disease (p < 0.001). Samples from healthy individuals were negative and only 5/12 infectious mononucleosis samples were positive. Real‐time quantitative PCR was subsequently employed to determine the concentration of EBV DNA present in serum; among positive samples the level ranged from 1 to 705 copies per 125 μl of serum. Post‐treatment samples from 5/14 cases with EBV‐associated HD contained detectable EBV DNA; analysis of this small group of cases suggests that positivity in post‐treatment samples correlates with risk factors indicative of a poor prognosis. Overall, our results are consistent with the notion that DNA from Reed‐Sternberg cells is present in the serum of HD patients, and further suggest that serum EBV should be evaluated as a prognostic marker. Int. J. Cancer (Pred. Oncol.) 84:442–448, 1999.

Screening for herpesvirus genomes in common acute lymphoblastic leukemia

There is epidemiological evidence that infection may play a role in the etiology of childhood leukemia in particular common B cell precursor acute lymphoblastic leukemia. A panel of 20 leukemic samples (panel 1) was examined for the presence of four lymphotropic herpesviruses using conventional molecular techniques. A second independent panel of 27 leukemic samples (panel 2), along with 28 control peripheral blood samples from children with other forms of cancer, was tested for the presence of the same four viruses using sensitive real-time quantitative PCR. While herpesvirus genomes were detected, they were present at very low levels; detection rates and levels were similar in the leukemic and control panels. In addition we surveyed 18 leukemic samples (five from panel 1, six from panel 2 and a further seven samples not previously analyzed) using a degenerate PCR assay capable of detecting the genomes of known herpesviruses plus putative new members of the family. No novel herpesvirus genomes were detected suggesting that a herpesvirus is unlikely to be etiologically involved as a transforming agent in common acute lymphoblastic leukemia.

JC and BK virus sequences are not detectable in leukaemic samples from children with common acute lymphoblastic leukaemia

Epidemiological evidence suggests that childhood leukaemia, and possibly common acute lymphoblastic leukaemia in particular, may have an infectious aetiology. Smith (1997 J Immunother: 89–100) recently suggested that the critical infectious event occurs during pregnancy, and identified the polyoma virus JC as a candidate agent. In the present study we investigated whether genomes from the JC virus, and closely related BK virus, could be detected in leukaemic cells. No positive results were obtained suggesting that JC virus is unlikely to play a direct role in leukaemogenesis.

Viruses and Hodgkin disease: No evidence of novel herpesviruses in non-EBV-associated lesions

The Epstein‐Barr virus (EBV) is associated with a proportion of cases of Hodgkin disease (HD) and this association is believed to be causal. Epidemiological studies suggest that an infectious agent is involved in the aetiology of young adult HD, however, cases in this age group are less likely to have EBV‐associated disease than cases diagnosed in early childhood or older adult years. Molecular studies have failed to find a consistent association between HD and other candidate viruses, and the aetiology of non‐EBV‐associated cases remains obscure. We looked for evidence of herpesvirus infection in samples of non‐EBV‐associated HD using a highly sensitive, degenerate PCR assay. Despite exhaustive sequence analysis of PCR products, no novel herpesviruses were identified. These results suggest that it is extremely unlikely that a novel herpesvirus is involved in the pathogenesis of non‐EBV‐associated HD.

Analysis of Epstein–Barr virus (EBV) nuclear antigen 1 subtypes in EBV-associated lymphomas from Brazil and the United Kingdom

EBNA-1 is the only viral protein consistently expressed in all cells latently infected by Epstein-Barr virus (EBV). There is a high frequency of sequence variation within functionally important domains of EBNA-1, with five subtypes identified. Individuals may be infected with multiple EBV strains (classified according to EBNA-1 subtype), but Burkitts lymphoma (BL) tumours carry a single subtype and exhibit some subtype preference. Subtype variation has also been related to geographical location. In the present study EBNA-1 polymorphisms were examined in a series of haematological malignancies from two distinct geographical regions, Brazil and the United Kingdom. Nucleotide sequence analysis of the carboxy-terminal region of EBNA-1 in 34 cases revealed six distinct sequences, some of which are novel. A new subtype, named V-Ala, was identified. EBNA-1 subtype in tumours differed markedly according to geographical location. In contrast to previous studies, we found evidence of EBNA-1 sequence variation within individual BL tumour samples.

Determination of HLA-A*02 antigen status in Hodgkin's disease and analysis of an HLA-A*02-restricted epitope of the Epstein-Barr virus LMP-2 protein.

There is good evidence for an association between Epstein‐Barr virus (EBV) and Hodgkins disease (HD). In approximately one‐third of cases, the EBV genome is detectable in Reed‐Sternberg (RS) cells and there is expression of the viral nuclear antigen EBNA‐1 and the latent membrane protein LMP‐1. Expression of LMP‐2 has been demonstrated at the mRNA level, and it is presumed that the protein is expressed alongside LMP‐1. The LMP‐2 protein is known to contain an epitope presented to cytotoxic T‐cells which is restricted through the HLA class I antigen A*0201 in healthy seropositive individuals. Since most HLA‐A*02‐positive Caucasians are HLA‐A*0201‐positive, it was hypothesized that HLA‐A*02‐positive individuals would be under‐represented among Caucasians with EBV‐associated HD. HLA‐A*02 status was determined, using flow cytometry and/or the polymerase chain reaction, for 276 individuals including 176 cases of HD. There was no significant difference between the frequency of HLA‐A*02 positivity in HD cases and controls, and between EBV‐associated and non‐associated cases of HD. The A*02 alleles of 14 cases of EBV‐associated HD were further subtyped using nested PCR; all except one case were found to be A*0201‐positive. We therefore investigated whether there was any evidence for mutation of the epitope representing amino acids 426–434 of LMP‐2a which is restricted through HLA‐A*0201. In 10/11 cases the nucleotide sequence encoding this epitope was identical to the published sequence; in the remaining case there was a mutation which would not be expected to alter the conformation of the epitope. Overall, our data suggest that other mechanisms of immune escape must be operative in EBV‐associated HD. Int. J. Cancer 72:614–618, 1997.

The response of Physical Science post‐graduates to training courses and the connection to their PhD studies

Training in both employability and discipline‐specific skills has been provided and expanded over a number of years for post‐graduate research students, (PGRs) in the Faculty of Physical Sciences administered by the Physical Sciences Graduate School (PSGS) at the University of Glasgow. This project explored the training provided in 2005/06 with a view to further developing a programme that students and faculty alike consider appropriate, timely and developmental for the needs of research students. The training provided by the PSGS had grown over a number of years in response to suggestions from academic staff in the Faculty of Physical Sciences. Data were collected from Postgraduate Research students (PGRs) from all the stages of the 3 year PhD process to enable a complete map of views to emerge. In particular, the way PGR students perceive the training they undergo in relation to their core PhD research and career progression was examined. The students in our study also identified clearly where they perceived they were developing such transferable skills, and training sessions are not seen as the sole or even major source; the research group itself would appear to play a major role. The authors believe the finding could inform the provision of PGR training in other UK institutions.

The variation in academics’ experiences of teaching in an intense study centre compared with their traditional university settings.

This article explores how teachers’ experiences of teaching accelerated courses in a residential setting compared with their experiences of teaching in their traditional contexts. It looks at how teachers responded to the opportunities the accelerated format provided and how this caused them to revisit not only what they taught, but how they taught. Teaching in an intense residential setting also allowed teachers to be more aware of their students as learners and caused them to revisit the purpose of higher education. Suggestions are made for how accelerated courses could be seen as part of an academic year as they offer advantages to both teacher and student.

Supporting the development of undergraduates’ experimental design skills and investigating their perceptions of project work

Abstract Project work represents a significant component of most Bioscience degrees. Conscious that students are not necessarily given adequate preparation for their final year project, we have investigated two core elements in the 3rd year of a 4-year Honours programme. One element, an investigative project on aspects of insect biology, has run for several years. The second, an Experimental Design course, was run for the first time in 2003, in order to support the development of key skills for bioscience graduates. The present paper describes a multi-stranded evaluation of these core elements. Significantly increased marks were seen in the Insect Projects in the years after the Experimental Design course was introduced. It is clear from our analysis that the students valued the Experimental Design course, believing that it had made them better able to design experiments and critique the experiments of others. We therefore conclude that the Experimental Design course is, at least in part, responsible for students’ improved performance in the Insect Projects. However, we do not believe that this is the only factor that has made the students engage with the Insect Projects more successfully than in previous years. From in-depth student interviews we identify a number of features of project work that are likely to enhance student engagement and motivation. Finally, we present some of the students’ conceptions of their prior experiences of experimental work that, we believe, give an indication of why students do not always value the opportunities to undertake experimental work that their courses offer them.