Development of a protein phosphatase-based assay for the detection of phosphatase inhibitors in crude whole cell and animal extracts

Diarrhetic shellfish poisoning (DSP) is a serious and globally widespread phytoplankton-related seafood illness. Although DSP is rarely life-threatening, it causes incapacitating diarrhea and vomiting with no known medical treatments. In addition, phytoplankton producing DSP toxins have been identified in temperate coastal waters worldwide, and their numbers may be increasing as a result of coastal eutrophication. The toxic effects of the major DSP toxins, okadaic acid and dinophysistoxin-1 (35-methylokadaic acid), appear to originate from their inhibitory activity against a family of structurally related serine/threonine protein phosphatases (PSPases). In particular, the inhibition of essential PSPases (e.g. PP1 and PP2A) has catastrophic consequences in most eukaryotic cells. Exploiting the potent inhibitory property of the DSP toxins, we have developed an enzyme-based assay (PP2A assay) capable of detecting both okadaic acid and dinophysis- toxin-1 in nanogram amounts. The assay employs purified PP2A, which has an extremely high affinity for both DSP toxins. This provides the PP2A assay with a level of sensitivity comparable to, or surpassing, that of most monoclonal antibody probes. To evaluate the PP2A assay as a means of detecting contaminated shellfish, a series of spike recovery experiments was conducted. The findings from these studies suggest that the PP2A assay has the potential for development into a rapid and relatively simple method for detecting PSPase inhibitors in crude extracts produced from shellfish.     

Changes to the ocular biota with time in extended- and daily-wear disposable contact lens use.

Gram-negative bacteria may play a role in the etiology of certain soft contact lens (SCL)-related diseases. Contact lens (CL) wear may modify the normal ocular biota, providing a more favorable environment for potential pathogens. This study reports temporal changes in ocular biota in daily-wear (DW) and extended-wear (EW) disposable SCL use in experienced and neophyte wearers. Lid margin and bulbar conjunctival biota were sampled prior to CL fitting in 26 previous DW SCL users, 18 previous EW SCL users, and 26 neophytes. Wearers were fitted with an etafilcon A CL in one eye and a polymacon CL in the fellow eye. Lenses were worn on a daily basis by the 26 previous DW SCL wearers and on an EW basis by the remaining 44 subjects. The ocular biota was further sampled after 1, 3, 6, 9, and 12 months of wear. The ocular biota consisted of coagulase-negative staphylococci, Corynebacterium spp., Micrococcus spp., and Propionibacterium spp. Potential pathogens were rarely isolated at baseline. No significant trend of increasing ocular colonization was shown for extended CL wear. Lid and conjunctival colonization increased with DW SCL use (P < 0.001), although this increase occurred for nonpathogenic species only. Fewer potential pathogens were isolated from DW SCL than from EW SCL users (P < 0.05). The lid margin consistently showed greater colonization than the conjunctiva and may be a source of potential pathogens during CL wear. Hydrogel CL wear appears to modify the ocular biota. An increased number of commensal organisms were present in DW SCL use. EW SCL use altered the spectrum of organisms isolated. These alterations may suppress the normal ocular defense mechanisms and may be relevant in the pathogenesis of CL-related disease.     

Changing perspectives in children hospitalized with poststreptococcal acute glomerulonephritis

Changing perspectives in 95 children with poststreptococcal acute glomerulonephritis (PSAGN) in our hospital between 1979 and 1988 are reported. Between 1961 and 1970 an average of 31±6.3 patients/year with PSAGN were treated and 70% had antecedent pyoderma. In the present study antecedent pharyngitis was observed in 59 children und pyoderma in 36. In comparison to the decade ending in 1970 our data show: (1) a marked decline in the prevalence of PSAGN (P=<0.0005), (2) a predominance of antecedent pharyngeal infection (P=0.044), (3) a decline in urban and an increase in rural patients with PSAGN (P=0.0483); and in the last decade: (1) a predominance of antecedent pharyngeal infection in children over 6 years of age (P=0.0009) and (2) a predominance of antecedent pyoderma in black children (P=0.0004).     

Case Report: Giant choledochal cyst

We report an adult female with a rare giant choledochal cyst. The patient presented following a normal pregnancy with the classical triad of an abdominal mass associated with jaundice and right upper quadrant abdominal pain. The cyst was excised using an intramural technique and biliary reconstruction achieved with a Roux-en-Y hepaticojejunostomy. Our patient has remained well with no evidence of malignancy over a 12 year review period. The aetiology and current management of this condition are discussed.     

Malignant melanoma in relation to moles, pigmentation, and exposure to fluorescent and other lighting sources

Interviews were performed on 83 patients with malignant melanoma, being 74% of all new NHS patients over a 33 month period who were resident in a defined area of Nottingham, and on age and sex matched controls chosen from all outpatients and inpatients of the same hospitals with the same area of residence. Significantly increased risks of melanoma were found in subjects with 3 or more raised moles on the upper arms (relative risk = 17.0), in association with heavy freckling of the face and arms, and with a tendency to sunburn easily and tan poorly, these factors having independent effects. While no significant and consistent association with exposure to fluorescent light was seen, the observed risks were higher in subjects with greater exposure, and higher in association with exposure to undiffused than to diffused light. Cases had a significantly greater number of hours' exposure to undiffused light than did controls. The associations with fluorescent light exposure were stronger when based on interview data than on a subsequent postal questionnaire. Twenty-one cases and 11 controls reported exposure to unusual occupational lighting sources which may have had an ultraviolet component; these included various intense lighting sources and lamps used in printing and dyeline copying.     

Prospective study of factors predicting uptake of smoking in adolescents.

Risk factors for the uptake of cigarette smoking were examined prospectively in 2159 non-smoking secondary schoolchildren aged 11-13 who participated in a survey in 1983 and were followed up 30 months later, by which time 35 per cent had taken up smoking. In a multivariate logistic model, the strongest predictors to emerge were prior experimentation with cigarettes and sex, with more girls (41%) than boys (30%) starting to smoke. Other predictors of taking up smoking were being uncertain about smoking in the future, reporting having been drunk, having a boy or girl friend, believing teachers and friends would not mind if they took up smoking, and giving lower estimates of prevalence of smoking among teachers. Parental smoking behaviour and attitudes, beliefs about the effects of smoking on health, opinions about smoking and perceived strictness of parents did not predict take up of smoking when other variables were controlled for. The odds of taking up smoking varied from 0.24 (risk = 0.19) for a child with the most favourable combination of risk factors to 3.49 (risk = 0.78) for a child with the worst prognosis. These results differ from those of many cross sectional studies and hence indicate the importance of a prospective approach to this type of research.