An in vivo examination of the stability of venom from the Australian box jellyfish Chironex fleckeri.

We have previously characterised the pharmacological activity of a number of jellyfish venoms with a particular emphasis on the profound cardiovascular effects. It has been suggested that jellyfish venoms are difficult to work with and are sensitive to pH, temperature and chemical changes. The current study aimed to examine the working parameters of the venom of the Australian box jellyfish Chironex fleckeri to enable fractionation and isolation of the toxins with cardiovascular activity. C. fleckeri venom was made up fresh each day and subjected to a number of different environments (i.e. a pH range of 5-9 and a temperature range of 4-30 degrees C). In addition, the effect of freeze drying and reconstituting the venom was investigated. Venom (50 microg/kg, i.v.) produced a transient hypertensive response followed by cardiovascular collapse in anaesthetised rats. This biphasic response was not significantly effected by preparation of the venom at a pH of 5, 7 or 9. Similarly, venom (50 microg/kg, i.v.) did not display a loss of activity when exposed to temperatures of 4, 20 or 30 degrees C for 1.5h. However, the cardiovascular activity was abolished by boiling the venom. Freeze drying, and then reconstituting, the venom did not significantly affect its cardiovascular activity. However, repeated freeze drying and reconstituting of extracted venom resulted in a significantly loss of activity. This study provides a more detailed knowledge of the parameters in which C. fleckeri venom can be used and, while supporting some previous studies, contradicts some of the perceived problems of working with the venom.

Epidemiology of insulin dependent diabetes before age 20 in Wisconsin, with particular reference to seasonality.

Physicians in the State of Wisconsin were contacted by mail and asked to report all cases of diabetes in patients under 20 yr diagnosed between 1 July 1982 and 30 June 1984 in order to study factors associated with seasonality in insulin-dependent diabetes mellitus (IDDM). Wisconsin's population is fairly homogeneous and is primarily middle socioeconomic class, small-town or rural, and of northern European Caucasian descent. The incidence of IDDM in winter was higher than in summer during the first year of the study, similarly to results of other studies. However, there was no significant winter peak in diagnosis during the second year. When monthly incidence rates from both years were combined, the increased evidence of IDDM in winter vs. summer was evident in males, but not in females. There appeared to be a spike in the number of new cases of IDDM in the first year of the study which was more evident in males. Such a spike is consistent with spikes in the incidence of IDDM occurring about the same time in Europe and in North America. The percentage of patients with antibody titres to Coxsackie virus and mycoplasma pneumoniae diagnosed during the first winter's peak were equal to those in nondiabetic controls. The distributions of HLA DR types of patients diagnosed in winter were no different from diabetics diagnosed in other seasons. The distribution of HLA DR types (5% DR2, 55% DR3, 82% DR4 and 38% DR3DR4) were similar to those of other groups of Caucasian subjects with IDDM. Also similarly to other studies of IDDM, 14% of the patients had thyroid microsomal antibody titers. The results of this study support the previously-advanced idea that winter might precipitate overt carbohydrate intolerance in individuals in whom insulin cell destruction is already well established (Diabetes, 36, 265-268, 1987). If this is true, studies of seasonality in IDDM might not be informative about the causation of IDDM.

HLA-DR4 in insulin-dependent diabetic parents and their diabetic offspring: a clue to dominant inheritance.

Insulin-dependent diabetes mellitus (IDDM) susceptibility determinants are known to be associated with both HLA-DR3 and -DR4. We monitored the inheritance of HLA-DR alleles in 37 families in which IDDM affected one parent and at least one offspring in order to try to learn more about the modes of inheritance of IDDM determinants. Ninety-seven insulin-dependent diabetics whose parents did not have diabetes and 158 nondiabetics were used as control groups for estimates of DR allele frequencies in the overall diabetic and general populations. The proportion of diabetic parents who transmitted DR4 to diabetic offspring (78%) was significantly higher (P less than 0.001) than the gene frequency of DR4 in the overall diabetic population (43%). The proportion of nondiabetic parents who transmitted DR4 to diabetic offspring (22%) was not significantly different from the gene frequency of DR4 in the nondiabetic population (16%), but it was significantly lower (P less than 0.05) than the gene frequency in the overall IDDM population. These proportions suggest that inheritance of the DR4-associated IDDM susceptibility determinant is not recessive, because in recessive inheritance expression of a trait depends on each parent contributing a susceptibility determinant. The proportions of diabetic and nondiabetic parents who transmitted the DR allele associated with the susceptibility determinant would then equal one another. The transmission of predominantly DR4 from affected parents to affected offspring suggests that susceptibility to IDDM is inherited primarily via a single dose of a potent determinant associated with DR4, as in dominant inheritance. When DR3 was transmitted at all it was usually by the nondiabetic parent. Only 8% of diabetic parents transmitted DR3 but 35% of nondiabetic parents transmitted DR3. The proportion of nondiabetic parents who transmitted DR3 was similar to the gene frequency of DR3 in the overall diabetic population (29%), but it was significantly higher than the gene frequency of DR3 in the nondiabetic population (15%; P less than 0.005). The percentage of diabetic offspring with the genotype DR3DR4 (35%) was identical to the percentage of individuals in the overall IDDM population with this genotype (35%). Numerous population data indicate that the DR3DR4 genotype carries a higher relative risk for IDDM than any other genotype, which suggests synergism between the DR3- and DR4-associated determinants. The family data reported here support this synergism but suggest that the DR4-associated determinant can give substantial susceptibility independent of the DR3-associated determinant and that the DR3-associated determinant is often expressed as enhancing susceptibility in the presence of the dominant DR4- associated determinant.