Epidermal cell-shape regulation and subpopulation kinetics during butyrate-induced terminal maturation of normal and SV40-transformed human keratinocytes: epithelial models of differentiation therapy

Recent data indicate that malignant human epidermal cells may be appropriate targets for sodium butyrate (NaB)-mediated differentiation therapy. The response of pre- and post-crisis populations of SV40-transformed human keratinocytes (SVKs) to this differentiation-inducing agent was assessed, therefore, within the framework of NaB-directed normal human keratinocyte (NHK) maturation. NaB augmented cornified envelope (CE) production in NHK and pre-crisis SVK cultures; the time-course and efficiency of induced maturation were similar in the 2 cell systems. In NHKs, the percentage of amplifying ("B" substate) cells decreased with time in NaB correlating with increases in both "C" stage keratinocytes and CEs. The latter formed over one or 2 layers of nucleated basal-like cells. Inductions were accompanied by immediate cell cycle blocks (in both the G1 and G2/M phases), reorganization within the actin cytoskeleton, and transient early increases in cellular actin content. Increased NHK and pre-crisis SVK cytoskeletal-associated actin reached a maximum approximately 48 hr after NaB addition and preceded development of CEs. The CE precursors, thus, probably reside in the "B" substate. Post-crisis SVKs, in contrast, were refractive to NaB-induced terminal maturation or cell-cycle perturbation, failed to initiate actin filament rearrangements, and retained a basal cell-like phenotype. Stable transformation of human SVKs in post-crisis phase, therefore, appears to be associated with loss of maturation "competence" within the "B" keratinocyte subpopulation.     

Detection of antibodies to caprine arthritis-encephalitis virus using recombinant gag proteins

Summary The coding sequences of the core proteins p17 and p28 of caprine arthritis-encephalitis virus (CAEV) were amplified using the polymerase chain reaction and cloned into the plasmid expression vector p-GEX-2T. Both p17 and p28 were expressed as fusion proteins with glutathione S-transferase. The recombinant proteins were affinity purified from induced bacterial lysates using glutathione-agarose beads. The purified proteins were used in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies against CAEV in goat sera and milk samples. Three different ELISA tests were developed based on p17, p28 or the combination of these two recombinant proteins (p17+p28). A comparison was made to an ELISA based on purified whole virus particles and to agar immunodiffusion test (AGID). Sera with conflicting results in the different ELISA tests were examined by Western blotting. There was a high correlation between the ELISA tests based on p17+p28 recombinant proteins and whole virus ELISA, with an estimated value of 0.92. Only 72–75% of the sera that tested positive in these two ELISA tests were positive in AGID. Antibodies to CAEV were detected in significantly more animals when serum samples were tested compared to milk samples. Based on the time and materials required to prepare the reagents, the recombinant based ELISA test was less expensive than the whole virus ELISA.     

A season of football injuries

All rugby and soccer players presenting to the Accident & Emergency department during the football season 1992-1993 (a total of 871) were prospectively studied to compare the injuries sustained in the two sports. The nature and site of injury, treatment required, age, fitness, experience and position of the player, situation giving rise to injury, and medical attention at the grounds were all analysed. The results show that rugby and soccer players had the same number of injuries, and while there were some differences in the nature of the injuries, there was no difference in overall severity. Rugby flankers and soccer goalkeepers are particularly at risk. Competitive matches produce more injuries than training sessions. Experience or fitness did not appear to be a factor and 45% of rugby injuries and 15% of soccer injuries were from school matches. Law changes (e.g. the rugby scrum and the use of gum-shields) have reduced some injuries, but other areas (e.g. jumping for the ball in soccer, rucks and mauls in rugby) also warrant consideration. There was one death, but no spinal cord injuries. Medical attention at the grounds was limited. Rugby injuries, therefore, do not appear to be more numerous or severe than soccer injuries. Law changes have been of benefit but they need to be enforced and perhaps more should be considered. Medical attention at sports grounds could be improved and Registers of injuries kept by the sporting bodies would be of benefit.     

Release and actions of adenosine in the central nervous system

Adenosine is released from active neurons into the extracellular fluid at a concentration of about 1mol/l. Neither the precise cellular origin nor the biochemical form of release has been firmly established, though the nucleotide is probably released partly directly, as a result of raised intracellular levels, and partly as nucleotides, which are subsequently hydrolysed. Once in the extracellular medium, adenosine markedly inhibits the release of excitatory neurotransmitters and modulatory peptides and has direct inhibitory effects on postsynaptic excitability via A₁ receptors. A population of A₂ receptors may mediate depolarization and enhanced transmitter release. Adenosine also modulates neuronal sensitivity to acetylcholine and catecholarnines, all these effects probably contributing to the behavioural changes observed in conscious animals. As a result of their many actions, adenosine analogues are being intensively investigated for use as anticonvulsant, anxiolytic, and neuroprotective agents.     

Awake oral fibreoptic intubation for caesarean section

A 35-week pregnant patient with ankylosing spondylitis and a known previous failed intubation required an elective caesarean section for intrauterine growth retardation. Regional anaesthesia was prevented by extensive spinal fusion. The anaesthetic management involved an awake oral fibreoptic intubation followed by induction and maintenance of general anaesthesia allowing delivery of a live infant without harm to the mother.     

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