The role of hydrogen peroxide in the in vitro cytotoxicity of 3-hydroxykynurenine

Previous studies have indicated that the generation of H₂O₂ may be a key step in the mechanism mediating the in vitro cytotoxicity of 3-hydroxykynurenine (3HK). An exposure protocol resulting in a delayed toxicity was utilized in order to further examine the role of H₂O₂ in the in vitro toxicity of 3HK in a neural hybrid cell line. 3HK-induced cell lysis was significantly attenuated by administration of catalase after termination of 3HK exposure and was abolished when intracellular peroxidase activity was elevated by pretreatment of cultures with horseradish peroxidase. In addition, a dose-dependent attenuation of 3HK toxicity was observed when cultures were exposed to 3HK in the presence of the iron chelator, desferrioxamine (DFO). Pretreatment with DFO also resulted in a significant attenuation of 3HK toxicity. These data suggest a direct role for H₂O₂ and metal ions in the cytotoxic action of 3HK and indicate that cell lysis results from the intracellular accumulation of toxic levels of H₂O₂.     

Clinical, laboratory, and genetic investigations of hypophosphatasia: support for autosomal dominant inheritance with homozygous lethality

This article presents detailed clinical and laboratory investigations of six hypophosphatasia kindreds. Serum alkaline phosphatase and urinary phosphoethanolamine comparisons between the affected population and a normal control population demonstrate these parameters routinely identify the heterozygous individual when age and sex variations are accounted for. Using clinical data from the kindred population and a detailed review of the literature, the type and frequency of clinical findings for both the homozygous and heterozygous genotype are enumerated. The clinical and biochemical phenotypes were subjected to segregation analysis. When the results of these analyses are viewed in light of their mathematical limitations and the genetic precepts of autosomal dominant and recessive inheritance, hypophosphatasia is best described as an autosomal dominant disorder with 85% penetrance and homozygous lethality.     

Influence of phospholipid asymmetry on fusion between large unilamellar vesicles.

The ability of lipid asymmetry to regulate Ca(2+)-stimulated fusion between large unilamellar vesicles has been investigated. It is shown that for 100-nm-diameter LUVs composed of dioleoylphosphatidylcholine, dioleoylphosphatidylethanolamine, phosphatidylinositol, and dioleoylphosphatidic acid (DOPC/DOPE/PI/DOPA; 25:60:5:10) rapid and essentially complete fusion is observed by fluorescent resonance energy transfer techniques when Ca2+ (8 mM) is added. Alternatively, for LUVs with the same lipid composition but when DOPA was sequestered to the inner monolayer by incubation in the presence of a pH gradient (interior basic), little or no fusion is observed on addition of Ca2+. It is shown that the extent of Ca(2+)-induced fusion correlates with the amount of exterior DOPA. Further, it is shown that LUVs containing only 2.5 mol % DOPA, but where all the DOPA is in the outer monolayer, can be induced to fuse to the same extent and with the same rate as LUVs containing 5 mol % DOPA. These results strongly support a regulatory role for lipid asymmetry in membrane fusion and indicate that the fusogenic tendencies of lipid bilayers are largely determined by the properties of the monolayers proximate to the fusion interface.     

Measurements of buoyancy for some Antarctic notothenioid fishes from the South Shetland Islands

Buoyancy was measured for 258 specimens representing 13 species of adult and sub-adult nototheniids, bathydraconids, and channichthyids from the South Shetland Islands. Measurements were expressed as percentage buoyancy (%B)=W_water/W_air᎒². There were no neutrally buoyant species and mean values for %B were 3.07-6.11%, with channichthyids at the low end and benthic nototheniids and bathydraconids at the high end. All species showed an ontogenetic decrease in %B with increasing body weight. With the exception of Champsocephalus gunnari, there was no sexual dimorphism in %B within this sample. With some exceptions, values for %B were consistent with life-history information. Sub-adult Dissostichus mawsoni were not neutrally buoyant, as are large adults. Notothenia rossii had a significantly lower %B than closely related N. coriiceps. Benthic Gobionotothen gibberifrons had a lower %B than semipelagic Lepidonotothen larseni. Although they exhibit some diversification in life history, the four channichthyids in the sample were similar in %B. Neutral buoyancy is rare in notothenioids and may be confined to a single nototheniid clade.     

Monitoring ovarian function in marmosets and tamarins by the measurement of urinary estrogen metabolites

Practical aspects of urinary estrogen analysis were considered with regard to establishing simple and reliable methods for monitoring ovarian function in marmosets and tamarins. Changes in the hormone:creatinine ratio in small volumes of urine from the common marmoset were significantly correlated with changes in 24-h excretion. Comparison of the metabolism and excretion of estrogens during the ovarian cycle in the common marmoset and cottontop tamarin revealed interesting species differences. High concentrations of conjugated estrone were measured in marmoset plasma, but estradiol 17β was the predominant estrogen in urine. In contrast, estrone was the most abundant estrogen measured in tamarin urine. Both species excreted very little estriol. Sulfates and glucuronides were present in urine in similar proportions before ovulation in the marmoset, although after ovulation sulfates were the more abundant. Conversely, most of the estrogens in tamarin urine appeared to be conjugated as glucuronides. Direct assay for estrone sulfate was applied to the measurement of urinary estrogen excretion during the ovarian cycle in a marmoset. The results compared well with those for total estradiol 17β after hydrolysis and ether extraction. The use of direct assays for conjugated estrogens in small volumes of urine is suggested as a practical method for monitoring ovarian function in marmosets and tamarins.     

Buoyancy studies and microscopy of skin and subdermal extracellular matrix of the antarctic snailfish,Paraliparis devriesi

The Antarctic snailfish Paraliparis devriesi (Liparididae) occupies an epibenthic habitat at a depth of 500–650 m in the subzero waters of McMurdo Sound, Antarctica. Although lacking a swim bladder, this species is neutrally buoyant through the combined effects of reduced skeletal ossification and expansion of a watery gelatinous subdermal extracellular matrix (SECM). The SECM serves as a low density buoyancy agent. It comprises a mean of 33.8% of the body weight, the largest known proportion of any adult fish. The SECM is loose connective tissue dominated by ground substance consisting of glycosaminoglycans, especially hyaluronic acid, and immobilized water. Although the SECM is 97% water, elevated levels of NaCl provide an osmotic strength greater than that of other body fluids. Only small amounts of antifreeze compounds have been identified in P. devriesi; therefore, freezing avoidance may result from the combined effects of antifreezes and the elevated osmolality of body fluids. The skin overlying the SECM is thin (85–200 μm) and loose, and unlike most other fishes, the epidermis is several times thicker than the dermis. The midepidermis, has a distinctive layer of vacuolated club cells of unknown function. Light and electron microscopy indicate that the skin is unspecialized for protection against entry of ice. © 1994 Wiley-Liss, Inc.