Enhanced Survival of the Cyanobacterium Oscillatoria terebriformis in Darkness under Anaerobic Conditions

Oscillatoria terebriformis, a thermophilic cyanobacterium, maintained viability in darkness under anaerobic conditions by fermenting exogenous glucose or fructose to lactic acid. The time period of survival was greatly extended when the environmental redox potential was lowered by the addition of sodium thioglycolate or titanium(III) citrate. When exposed to aerobic conditions in darkness, many trichomes underwent lysis in 6 h, and death of all cells occurred in 2 to 3 days. The endogenous aerobic respiration rate was high, and the limited dark aerobic survival period appeared to be due to depletion of stored glycogen. Fructose or glucose did not support or increase aerobic respiration in darkness or lengthen aerobic survival time. Enhanced survival of O. terebriformis in darkness under anaerobic, reducing conditions correlates well with the natural nighttime position of this species within sulfide-rich microbial mats associated with hot springs of western North America.     

Freeze-Thawing of Aquaspirillum magnetotacticum Cells Selectively Releases Periplasmic Proteins

Cells of the gram-negative bacterium Aquaspirillum magnetotacticum, when suspended in buffer and freeze-thawed, produced pinkish orange supernatant fluid. The fluid contained ≤2.0% of total extractable outer membrane component 2-keto-3-deoxyoctonate or of the cytoplasmic membrane marker succinic dehydrogenase. Electrophoretic banding patterns and difference spectra of proteins and hemoproteins released by freeze-thawing cells were distinct from those of membrane-associated substances and similar to those of periplasmic substances obtained by applying conventional fractionation methods to this organism.     

Abnormal subcellular distribution of β-glucuronidase in mice with a genetic alteration in enzyme structure

Liver -glucuronidase is structurally altered in inbred strain PAC so that a peptide subunit with a more basic isoelectric point, GUS-SN, is produced. This allele of -glucuronidase was transferred to strain C57BL/6J by 12 backcross matings to form the congenic line B6 · PAC-Gus ⁿ. Liver -glucuronidase activity was halved in males of the congenic strain compared to normal males. The lowered activity was specifically accounted for by a decrease in the lysosomal component. There was no alteration in the concentration of microsomal activity. This alteration in the subcellular distribution of -glucuronidase in Gus ⁿ/Gus ⁿ mice was confirmed by two independent gel electrophoretic systems which separate microsomal and lysosomal components. -Glucuronidase activity was likewise approximately halved in mutant spleen, lung, and brain, organs which contain exclusively or predominantly lysosomal -glucuronidase. The loss of liver lysosomal -glucuronidase activity was shown by immunotitration to be due to a decrease in the number of -glucuronidase molecules in lysosomes of the congenic strain. The Gus ⁿ structural alteration likely causes the lowered lysosomal -glucuronidase activity since the two traits remain in congenic animals. Heterozygous Gus ⁿ/Gus ^b animals had intermediate levels of liver -glucuronidase. Also, the effect was specific, in that three other lysosomal enzymes were not reproducibly lower in Gus ⁿ/Gus ⁿ mice. Gus ⁿ is, therefore, an unusual example of a mutation which causes a change in the subcellular distribution of a two-site enzyme.This work was supported by National Institutes of Health Grants GM-33559 and GM-33160 and National Science Foundation Grant PCM-8215808.     

Microheterogeneity of agonist and antagonist glucocorticoid receptor complexes detected by isoelectric focusing and modifications induced by receptor activation

Rat-liver glucocorticoid receptor was incubated with either [3H]triamcinolone acetonide or [3H]RU 486, a well known antiglucocorticoid. Once formed, the steroid-receptor complexes were analyzed by isoelectric focusing in agarose gel slabs. A careful slicing of the receptor tracks revealed the presence of three distinct radioactive peaks focused at the following pI values: 5.3 +/- 0.2 (n = 17) and 4.4 +/- 0.1 (n = 17). All these peaks correspond with receptor isoforms as suggested by control experiments. The receptor state was analyzed after focusing by a chromatographic assay on DNA-cellulose, DEAE-trisacryl and hydroxyapatite minicolumns. The peak of pI 4.4 apparently corresponded to the non-transformed receptor and was greatly stabilized in the presence of RU 486, whereas the peaks of pI 4.8 and 5.3 were probably made of transformed receptor and meroreceptor. These results were confirmed by autoradiographic studies after isoelectric focusing of receptor molecules covalently labelled with [3H]dexamethasone mesylate. Thus, the rat-liver glucocorticoid receptor appeared to be a rather acidic protein which became less acidic after transformation by heat, displaying a pI shift which was strongly reduced in case of steroid-receptor complexes formed with the antiglucocorticoid RU 486.     

AIDS, gays, and state coercion

Mohr argues that coercive government policies in response to the AIDS crisis are unjustified and pose a serious threat to the rights of homosexuals. AIDS presently can be transmitted only to those whose actions place them at risk. Paternalistic state coercion is warranted only when a person has diminished capacity or when necessary to prevent someone from ceasing to be an independent agent. Furthermore, an individual may regard sex as a central value. Finally, using public health as a rationale for closing bathhouses or otherwise regulating sexual behavior is a step toward totalitarianism, because coercion is unwarranted when protection from a disease can be achieved by individual action and because the public good served by reducing the size of the pool of AIDS-exposed people is outweighed by the unjust discrimination involved in using coercive measures that affect only certain individuals.     

Effects of pre-treatment with aflatoxin on a second aflatoxin treatment in guinea pigs

Two-hundred guinea pigs, weighing approximately 500 grams each, were placed in 8 groups, 4 of which received 20 g/kg/day of partially purified aflatoxin for 7 days, followed by a 7 day recovery period. Paired groups then received 0,20,35 or 50 g/kg/day of partially purified aflatoxin for 21 days. Animals were sacrificed periodically from all groups and blood was drawn for chemical and immunologic analysis. Weight gains were recorded and histopathologic studies were done on all animals. Pretreatment did not protect guinea pigs from a second exposure, and in fact enhanced mortality and liver toxicity as determined by histopathology. Serum chemistries and immunologic parameters of guinea pigs dosed twice were less conclusive, as neither high nor low doses differed from guinea pigs treated once. Glycocholic acid concentrations were more sensitive than traditional enzymes (aspartate and alanine amino transferase, alkaline phosphatase) for indicating hepatotoxicity.     

Regression of blood vessels precedes cartilage differentiation during chick limb development

We have previously investigated distinct areas of vascular regression in the developing vascular system of the chick limb bud. Avascular areas appear in a characteristic spatial and temporal pattern, and are correlated with the position of developing cartilage. In the present study, we examined limb-bud sections which had been double labeled for endothelial cells and developing cartilage in order to determine the relationship between the appearance of cartilage and the disappearance of capillaries. Endothelial cells, which specifically take up acetylated low-density lipoprotein (acLDL), were labeled by intravenously injecting fluorescent acLDL (DiIacLDL) into chick embryos at Hamburger and Hamilton stages 26-30. Avascular zones, which correspond to the developing digits, were clearly visible within the fluorescently labeled distal vasculature. The same sections were labeled with monoclonal antibodies specific for cartilage. We found that progressing avascularity in the digital regions was followed by increased staining for cartilage antigens in the same areas. Zones of avascularity always developed earlier than morphologically and immunologically detectable cartilage in all planes of section and were always larger than the areas of cartilage. These results demonstrate that blood vessels disappear in predictable areas prior to the overt differentiation of cartilage.