Formation of compound I and compound II ferryl species in the reaction of hemoglobin I from Lucina pectinata with hydrogen peroxide

The formation of ferryl heme (Fe(IV) = O) species, i.e., compound I and compound II, has been identified as the main intermediates in heme protein peroxidative reactions. We report stopped-flow kinetic measurements which illustrate that the reaction of hemoglobin I (HbI) from Lucina pectinata with hydrogen peroxide produce ferryl intermediates compound I and compound II. Compound I appears relatively stable displaying an absorption at 648 nm. The rate constant value (k'(2)) for the conversion of compound I to compound II is 3.0 x 10(-2) s(-1), more than 100 times smaller than that reported for myoglobin. The rate constant value for the oxidation of the ferric heme (k'(12) + k'(13)) is 2.0 x 10(2) M(-1) s(-1). These values suggest an alternate route for the formation of compound II (by k'(13)) avoiding the step from compound I to compound II (k'(2)). In HbI from L. pectinata the stabilization of compound I is attribute to the unusual collection of amino acids residues (Q64, F29, F43, F68) in the heme pocket active site of the protein.     

ATM and the catalytic subunit of DNA-dependent protein kinase activate NF-kappaB through a common MEK/extracellular signal-regulated kinase/p90(rsk) signaling pathway in response to distinct forms of DNA damage

We have identified a novel pathway of ataxia telangiectasia mutated (ATM) and DNA-dependent protein kinase (DNA-PK) signaling that results in nuclear factor kappaB (NF-kappaB) activation and chemoresistance in response to DNA damage. We show that the anthracycline doxorubicin (DOX) and its congener N-benzyladriamycin (AD 288) selectively activate ATM and DNA-PK, respectively. Both ATM and DNA-PK promote sequential activation of the mitogen-activated protein kinase (MAPK)/p90(rsk) signaling cascade in a p53-independent fashion. In turn, p90(rsk) interacts with the IkappaB kinase 2 (IKK-2) catalytic subunit of IKK, thereby inducing NF-kappaB activity and cell survival. Collectively, our findings suggest that distinct members of the phosphatidylinositol kinase family activate a common prosurvival MAPK/IKK/NF-kappaB pathway that opposes the apoptotic response following DNA damage.     

Presence of Cochlodinium catenatum (Gymnodiniales: Gymnodiniaceae) in red tides of Bahía de Banderas, Mexican Pacific

The evolution of an ichthiotoxic algal bloom caused by the dinoflagellate Cochlodinium catenatum was studied from July to December 2000. The abnormal multiplication of this dinoflagellate occurred in the form of a discoloration spreading between a temperature and salinity interval of 25-32 degrees C and 33-35 ups, respectively. The density of C. catenatum reached 10 841 cells ml(-1). The event was observed in large areas of Banderas Bay affecting 13 fish species, whose massive killing was due to suffocation (gill obstruction and excessive mucus production). The human population around the area did not present respiratory affections or skin irritation. The C. catenatum measurements suggest a hologamic and heterothalic reproduction. Their morphological characteristics suggest that C. polykrikoides, C. heterolobatum and C. catenatum are the same species. It is estimated that the species could be a recent introduction in the Mexican Pacific.     

Discoloration of microalgal blooms as a bioindicating parameter in the Bahia de Mazatlán, México

In order to verify whether harmful algal blooms increased since 1979, a long-term project for the study of harmful algal blooms in Mazatlán Bay began 25 years ago (1979-2003). Observations were done daily and results indicate five trend types: (I) years without discoloration related to El Ni?o events; (II) years with days of sudden occurrence of discoloration, related to the release of accumulated energy after El Ni?o events; (III) years with similar number of discoloration events (days); (IV) years with a progressive decrease of the occurrence (days) of discoloration events; (V) years with gradual increase of discoloration events. The annual mean average of discoloration is of 25.3 days. Ninety-six discoloration events were recorded during year 2000, and represent the highest number of events in the last 25 years. Discoloration events are more common in March (174 days), harmful and toxic blooms included. This indicates continuity in the occurrence of the highest content of microalgae, and therefore, it is concluded that they play an important role for a better understanding of the environmental deterioration undergone by the coastal zone due to an increase of harmful and toxic microalgae blooms.     

Benzodiazepine-based selective inhibitors of mitochondrial F1F0 ATP hydrolase

A series of benzodiazepine-based inhibitors of mitochondrial F(1)F(0) ATP hydrolase were prepared and evaluated for their ability to selectively inhibit the enzyme in the forward direction. Compounds from this series showed excellent potency and selectivity for ATP hydrolase versus ATP synthase, suggesting a potentially beneficial profile useful for the treatment of ischemic heart disease.     

N-[1-Aryl-2-(1-imidazolo)ethyl]-guanidine derivatives as potent inhibitors of the bovine mitochondrial F1F0 ATP hydrolase

A series of substituted guanidine derivatives were prepared and evaluated as potent and selective inhibitors of mitochondrial F(1)F(0) ATP hydrolase. The initial thiourethane derived lead molecules possessed intriguing in vitro pharmacological profiles, though contained moieties considered non-drug-like. Analogue synthesis efforts led to compounds with maintained potency and superior physical properties. Small molecules in this series which potently and selectivity inhibit ATP hydrolase and not ATP synthase may have utility as cardioprotective agents.     

Schizosaccharomyces pombe Pmr1p is essential for cell wall integrity and is required for polarized cell growth and cytokinesis

The cps5-138 fission yeast mutant shows an abnormal lemon-like morphology at 28 degrees C in minimal medium and a lethal thermosensitive phenotype at 37 degrees C. Cell growth is completely inhibited at 28 degrees C in a Ca2+-free medium, in which the wild type is capable of growing normally. Under these conditions, actin patches become randomly distributed throughout the cell, and defects in septum formation and subsequent cytokinesis appear. The mutant cell is hypersensitive to the cell wall-digesting enzymatic complex Novozym234 even under permissive conditions. The gene SPBC31E1.02c, which complements all the mutant phenotypes described above, was cloned and codes for the Ca2+-ATPase homologue Pmr1p. The gene is not essential under optimal growth conditions but is required under conditions of low Ca2+ (<0.1 mM) or high temperature (>35 degrees C). The green fluorescent protein-tagged Cps5 proteins, which are expressed under physiological conditions (an integrated single copy with its own promoter in the cps5Delta strain), display a localization pattern typical of endoplasmic reticulum proteins. Biochemical analyses show that 1,3-beta-D-glucan synthase activity in the mutant is decreased to nearly half that of the wild type and that the mutant cell wall contains no detectable galactomannan when the cells are exposed to a Ca2+-free medium. The mutant acid phosphatase has an increased electrophoretic mobility, suggesting that incomplete protein glycosylation takes place in the mutant cells. These results indicate that S. pombe Pmr1p is essential for the maintenance of cell wall integrity and cytokinesis, possibly by allowing protein glycosylation and the polarized actin distribution to take place normally. Disruption and complementation analyses suggest that Pmr1p shares its function with a vacuolar Ca2+-ATPase homologue, Pmc1p (SPAPB2B4.04c), to prevent lethal activation of calcineurin for cell growth.     

Solution structure of ribosomal protein S28E from Methanobacterium thermoautotrophicum

The ribosomal protein S28E from the archaeon Methanobacterium thermoautotrophicum is a component of the 30S ribosomal subunit. Sequence homologs of S28E are found only in archaea and eukaryotes. Here we report the three-dimensional solution structure of S28E by NMR spectroscopy. S28E contains a globular region and a long C-terminal tail protruding from the core. The globular region consists of four antiparallel beta-strands that are arranged in a Greek-key topology. Unique features of S28E include an extended loop L2-3 that folds back onto the protein and a 12-residue charged C-terminal tail with no regular secondary structure and greater flexibility relative to the rest of the protein. The structural and surface resemblance to OB-fold family of proteins and the presence of highly conserved basic residues suggest that S28E may bind to RNA. A broad positively charged surface extending over one side of the beta-barrel and into the flexible C terminus may present a putative binding site for RNA.     

Synthesis and in vitro trypanocide activity of several polycyclic drimane-quinone derivatives

The Diels-Alder reaction between two polygodial-derived dienes and simple quinones to yield substituted naphtho- and anthraquinones, is described. The in vitro trypanocide activity for the series was determined. Two of the new compounds showed an activity ten and two times higher, respectively, than nifurtimox and benznidazole, the medicines of choice for the treatment of the acute Chagas' disease.     

Feeding habits of guanacos<Emphasis Type="Italic">Lama guanicoe</Emphasis> in the high Andes of north-central Chile

The guanacoLama guanicoe Muller, 1776 has a wide distribution along the Andes and Patagonia. We studied the feeding behaviour of a guanaco population that lives over 4100 m altitude in the Andes of north-central Chile. By contrasting the diet of guanacos during a dry year with that of a wet year and comparing it with the plant availabilities in the field, we tested the hypothesis that the guanaco is a generalist herbivore. We predicted that under such extreme habitat conditions guanacos should consume whatever plant species are available in the environment, especially in a dry year, when vegetation is scarcer. In addition, we compared its diet at three different age classes. We estimated the diet through the microhistological analysis of plant remains found in guanaco pellets collected during January of 1997 (ie after a dry year) and 1998 (ie after a wet year; 41 vs 495 mm, respectively). Then, we computed dietary preferences, food niche-breadth, and food-niche overlap between years and among age classes. Vegetation cover and plant species richness were higher during the wet than during the dry year. The most common plants in the environment were perennial graminoids and legumes. Contrary to our prediction, the guanaco preferred a few plant species, showing a relatively narrow diet breadth that changed little between years differing in plant abundances. The diet proportions differed among the three age classes, however. Our data indicate that at least in this high-elevation population, guanacos are selective and non-opportunistic herbivores. This specialized feeding behaviour is puzzling given the energetic demands of living in a harsh environment with low availabilities of resources. The hypothesis that this is due to the lower palatability of the plants not eaten, remains to be tested.