Peroxynitrite-Induced Cytotoxicity in PC12 Cells: Evidence for an Apoptotic Mechanism Differentially Modulated by Neurotrophic Factors

Abstract: Peroxynitrite is a powerful oxidant formed by the near-diffusion-limited reaction of nitric oxide with superoxide. Large doses of peroxynitrite (>2 m M ) resulted in rapid cell swelling and necrosis of undifferentiated PC12 cells. However, brief exposure to lower concentrations of peroxynitrite (EC₅₀ = 850 µ M ) initially (3–4 h) caused minimal damage to low-density cultures. By 8 h, cytoplasmic shrinkage with nuclear condensation and fragmentation became increasingly evident. After 24 h, 36% of peroxynitrite-treated cells demonstrated these features associated with apoptosis. In addition, 46% of peroxynitrite-treated cells demonstrated DNA fragmentation (by terminal-deoxynucleotide transferase-mediated dUTP-digoxigenin nick end-labeling) after 7 h, which was inhibited by posttreatment with the endonuclease inhibitor aurintricarboxylic acid. Serum starvation also resulted in apoptosis in control cells (23%), the percentage of which was not altered significantly by peroxynitrite treatment. Although peroxynitrite is known to be toxic to cells, the present study provides a first indication that peroxynitrite induces apoptosis. Furthermore, pretreatment of cells with nerve growth factor or insulin, but not epidermal growth factor, was protective against peroxynitrite-induced apoptosis. However, both acidic and basic fibroblast growth factors greatly increased peroxynitrite-initiated apoptosis, to 63 and 70%, respectively. Thus, specific trophic factors demonstrate differential regulation of peroxynitrite-induced apoptosis in vitro.     

Regression of hypoxic hypertension-induced changes in the elastic laminae of rat pulmonary arteries

Liu, S. Q. Regression of hypoxic hypertension-inducedchanges in the elastic laminae of rat pulmonary arteries.J. Appl. Physiol. 82(5):1677-1684, 1997.The elastic laminae of the pulmonary arteries(PAs) undergo a progressive structural change in hypoxic hypertension.This study focused on the reversibility of altered PA elastic laminaeof the rat due to hypoxic hypertension. The structure andcross-sectional area of the PA medial elastic laminae were examined byusing electron-microscopic and image-analytic approaches duringrecovery from 12 h and 10 days of hypoxic hypertension. At 12 h ofhypoxic hypertension, the elastic laminae, which appeared homogeneousin normal control animals, were reorganized into structures composed ofrandomly oriented filaments, with an increase in the cross-sectionalarea of 70%. At 10 days of hypoxic hypertension, the elastic laminaeappeared homogeneous in structure and normal in cross-sectional areadespite continuous exposure to hypoxia. During recovery from 12 h ofhypoxic hypertension, the medial elastic laminae regained theirhomogeneous structure and normal cross-sectional area afterday 2. During recovery from 10 days ofhypoxic hypertension, the medial elastic laminae changed from homogeneous to filamentous structures, with a progressively altered cross-sectional area that increased by 89% from recoveryday 0 to day10 and returned to the normal level onday 30. These changes were associatedwith alterations in the PA wall tensile stress. These results indicatedthat structural changes in the PA elastic laminae were reversible andthat the regression process depended on the duration of exposure tohypoxia, the state of the elastic laminae, and possibly the tensilestress level in the PA wall.

     

Comparative Cytochrome Oxidase and Superoxide Dismutase Analyses on Strains of Azotobacter vinelandii and Other Related Free-Living Nitrogen-Fixing Bacteria

Quantitative N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD) oxidase and superoxide dismutase (SOD) analyses were performed on representative organisms of the family Azotobacteraceae. Azotobacter vinelandii, Azotobacter chroococcum, Azotobacter paspali, and Derxia gummosa exhibited high quantitative TMPD oxidase activities, and their extracts possessed very active and electrophoretically homogeneous (single gel band) Fe-type SODs. Azomonas macrocytogenes extracts had similar single Fe-type SODs, and their cells exhibited no TMPD-dependent cytochrome oxidase activity. Nitrogen-fixing cells of Beijerinckia indica, Beijerinckia derxii, and Beijerinckia mobilis exhibited minimal TMPD oxidation capabilities (rates equivalent to the TMPD autooxidation reaction), and these extracts also possessed very active SODs but only of the Mn metallotype.     

Sesquiterpenes of thalloid liverworts of the genera Conocephalum,Lunularia, Metzgeria and Riccardia

Thalloid liverworts of orders Metzgeriales and Marchantiales elaborate essential oils distinguishable from those of the Jungermanniales by the absence of β-barbatene and anastreptene. Riccardia sinuata elaborates a novel tricyclic exomethylene sesquiterpene of as yet undetermined structure. Conocephalum conicum elaborates cadinene-type sesquiterpenes. β-Cadinene from the latter species is clearly enantiomeric to the same product from vascular plants.     

Notes on the morphology,ecology and life cycles of Fukaurahydra anthoformis and Hataia parva (Hydrozoa,Athecata)

Fukaurahydra anthoformis and Hataia parva are solitary athecate hydroids occurring in northern Japan. New information on the external morphology, nematocysts, ecology, and life cycles of these species is presented. It is noteworthy that H. parva bears stenoteles, which are generally not found among the families of Filifera. Neither species produces free medusae. The eggs are fertilized in the female gonophores, from which unciliated larvae are released. These larvae do not swim and soon attach to a substrate. After attachment the larvae become covered by a sheath to form cysts. The cysts rest on a substrate without any outer change for several months. As the water temperature drops in autumn to early winter the cysts begin to hatch, forming tiny polyps after the larva creeps out from the chitinous sheath. Cyst formation proves to be common also in other solitary hydroids, most of which are inhabitants of cool or cold waters.     

与胎儿珠蛋白基因持续表达有关的(A_γδβ)°—地贫纯合子及杂合子的分子鉴别

我们分子鉴别了一个缺失型中国(A_γδβ)°-地贫家系。先证者为这一缺失的纯合子,具有中度贫血症状。家系的另五个成员均为这一缺失的杂合子,其胎儿血红蛋白(HbF)为16—21％,接近或达到HPFH杂合子的HbF水平,并且几乎不表现贫血症状。限制性内切酶图谱分析证明了β-珠蛋白基因簇内的DNA顺序缺失,缺失的5′端点位于Aγ基因IVSⅡ内,3′端点在β-珠蛋白基因下游区远端,距HPFH-2的3′缺失端点上游区约11kb。缺失的总长度约为80kb。本文讨论了这一缺失导致胎儿血红蛋白在成人中持续活跃表达的可能机制。