A B-cell targeting virus disrupts potentially protective genomic methylation patterns in lymphoid tissue by increasing global 5-hydroxymethylcytosine levels

The mechanisms by which viruses modulate the immune system include changes in host genomic methylation. 5-hydroxymethylcytosine (5hmC) is the catalytic product of the Tet (Ten-11 translocation) family of enzymes and may serve as an intermediate of DNA demethylation. Recent reports suggest that 5hmC may confer consequences on cellular events including the pathogenesis of disease; in order to explore this possibility further we investigated both 5-methylcytosine (5mC) and 5hmC levels in healthy and diseased chicken bursas of Fabricius. We discovered that embryonic B-cells have high 5mC content while 5hmC decreases during bursa development. We propose that a high 5mC level protects from the mutagenic activity of the B-cell antibody diversifying enzyme activation induced deaminase (AID). In support of this view, AID mRNA increases significantly within the developing bursa from embryonic to post hatch stages while mRNAs that encode Tet family members 1 and 2 reduce over the same period. Moreover, our data revealed that infectious bursal disease virus (IBDV) disrupts this genomic methylation pattern causing a global increase in 5hmC levels in a mechanism that may involve increased Tet 1 and 2 mRNAs. To our knowledge this is the first time that a viral infection has been observed to cause global increases in genomic 5hmC within infected host tissues, underlining a mechanism that may involve the induction of B-cell genomic instability and cell death to facilitate viral egress.     

Highly recurrent BRAF p.V595E mutation in canine papillary oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is the most common oral epithelial malignancy in dogs. It exhibits locally aggressive biological behaviour with the potential to metastasize, and a reported 1-year survival rate of 0% when left untreated. Expression studies suggest that aberrant MAPK signalling plays a key role in canine OSCC tumorigenesis, which is consistent with BRAF and HRAS MAPK-activating mutations reported in some tumours. Several morphological subtypes of canine OSCC have been described, with papillary, conventional, and basaloid as the most common patterns. We hypothesized that mutational differences may underlie these phenotypic variations. In this study, targeted Sanger sequencing and restriction fragment length polymorphism assays demonstrate that up to 85.7% of canine papillary OSCC (n = 14) harbour a BRAF p.V595E mutation. Assessment of neoplastic epithelial cell proliferation using Ki67 immunolabelling (n = 10) confirmed a relatively high proliferation activity, consistent with their known aggressive clinical behaviour. These findings underscore a consistent genetic feature of canine papillary OSCC and provide a basis for the development of novel diagnostic and targeted therapeutic approaches that can improve the quality of veterinary care.     

Current Techniques for Evaluation of Spinal Cord Injury

Spinal cord injuries have been evaluated in the past chiefly on the basis of subjective tests. New electronic techniques are now coming into clinical use which allow objective testing of afferent and efferent conduction through long spinal cord tracts. With continued research, it is probable that diagnosis and prognosis of spinal cord injuries can be made with a high degree of confidence.     

Pathology in the medical curriculum in the United Kingdom

In the United Kingdom, the medical students of yesteryear undertook a detailed pathology course, often with its own major examinations. This contributed to the increasing overload of factual information that was common in medical curricula. Following General Medical Council reports on medical education, pathology is now more fully integrated into clinical work and into problem-based learning, albeit with greatly reduced contact time with pathologists. Teaching methods have shifted from didactic presentations, such as lectures, to more varied methods, with emphasis on self-access and interactivity. Technological advances have brought well-illustrated textbooks, computer-assisted learning programs, Web resources, and electronic communication. There has been a decline in the role of museum specimens, microscopy, and, partly following a worldwide trend and partly following adverse media publicity, the traditional autopsy. Assessment methods have evolved, with integration of pathology into wider examinations. The greater drive toward increasing funding through the Research Assessment Exercise has had an adverse effect on teaching throughout universities. Despite these setbacks, the importance of pathology as a clinical discipline will ensure its continued role in medical education.     

THE EFFECTS OF INTRA-ARTICULAR ANESTHESIA ON SOFT TISSUE-AND BONE-PHASE SCINTIGRAPHY IN THE HORSE

Soft tissue- and bone-phase scintigrams were acquired from 4 normal horses before and over a 14-day period after metacarpophalangeal, antebrachiocarpal, tarsocrural and tarsometatarsal joint blocks. Images were evaluated subjectively and quantitatively for increased activity in these regions. The antebrachiocarpal block resulted in obvious focal accumulation of activity on soft tissue-phase scintigrams. This increased activity was greatest 2 to 4 days postanesthesia and persisted up to 14 days postanesthesia. On quantitative analysis of soft tissue-phase images, similar trends were noted after the metacarpophalangeal and tarsocrural blocks, but these increases were relatively mild and were not evident on subjective evaluation of the images. Abnormal soft tissue-phase activity was not associated with the tarsometatarsal block. On bone-phase scintigrams, increased activity was not present following any of these joint blocks.     

Cloning, expression and characterization of monkey (Macaca fascicularis) CD137

CD137 plays an important role as a co-stimulatory molecule in activated T cells. Agonistic CD137 specific antibodies have been investigated as therapeutic agents to promote tumor-specific immune responses by direct activation of T cells. As part of the pre-clinical pharmacological evaluation of cynomolgus monkeys, monkey CD137 was cloned and characterized. The deduced amino acid sequence encoded a full-length gene of 254 amino acids 95% identical to human CD137. Sequence variants identified in monkey CD137 include four splice variants lacking the transmembrane domain. These variants were detectable in human including two previously unreported variants. Two missense single nucleotide polymorphisms were detected present in 42 and 50% of 36 monkeys tested. In both monkey and human, mRNA expression of full-length CD137 and splice variants were significantly increased in peripheral blood mononuclear cells (PBMCs) upon stimulation by anti-CD3 antibodies. Recombinant monkey CD137 protein was bound with high affinity by an agonistic anti-human CD137 antibody but not by an anti-mouse CD137 antibody. In summary, compared to human, monkey CD137 showed distinct extracellular domain amino acid sequence and sequence polymorphisms. Thus, antibodies directed against epitopes in this extracellular domain could have differences in pharmacologic activity between cynomolgus monkeys and human or across individual cynomolgus monkeys.