Nitroarginine,an inhibitor of nitric oxide synthetase,attenuates ammonia toxicity and ammonia-induced alterations in brain metabolism

We have proposed that acute ammonia toxicity is mediated by activation of the N-methyl-D-aspartate type of glutamate receptors. MK-801, a selective antagonist of these receptors, prevents death of animals induced by acute ammonia intoxication as well as ammonia-induced depletion of ATP. It seems therefore that, following activation of the N-methyl-D-aspartate receptors, the subsequent events in ammonia toxicity should be similar to those involved in glutamate neurotoxicity. As it has been shown that inhibitors of nitric oxide synthetase such as nitroargnine prevent glutamate toxicity, we have tested whether nitroarginine prevents ammonia toxicity and ammonia-induced alterations in brain energy and ammonia metabolites. It is shown that nitroarginine prevents partially (50%), but significantly death of mice induced by acute ammonia intoxication. Nitroarginine also prevents partially ammonia-induced depletion of brain ATP. It also prevents completely the rise in glucose and pyruvate and partially that in lactate. Injection of nitroarginine alone, in the absence of ammonia, induces a remarkable accumulation of glutamine and a decrease in glutamate. The results reported indicate that nitroarginine attenuates acute ammonia toxicity and ammonia-induced alterations in brain energy metabolites. The effects of MK-801 and of nitroarginine are different, suggesting that ammonia can induce nitric oxide synthetase by mechanisms other than activation of N-methyl-D-aspartate receptors.     

Airborne pollen concentrations,solid particle content in the air and allergy symptoms in Córdoba (Spain)

During 1991 and 1992, hourly measurements were taken of solid particulates (size, 22–25 μm).Olea and grass pollen and spores. The results were then compared with symptoms recorded in patients consulting the Allergy Unit during the period of highest pollen concentrations (April–June). The spore-trap slides were studied by spectrophotometry in order to analyse the total solid particulates content in the air as a possible synergic agent of the pollen. This allowed the volume of material present in the air to be determined and expressed as a percentage of total optical density (OD). Allergy symptom data were obtained from the study of subjective clinical records completed by patients, who were required to note down the severity of a selected series of symptoms every 4 h using a scale from 0–3. The results showed a clear link during the season between hourly peaks and pollen concentrations, and the different patterns of response associated with the dominant pollen types. An attempt has been made to determine whether a relationship exists between the increment of the optical density and the symptomatic response. The positive relationship encountered seems to indicate that a synergic agent takes an active part in the effects produced by pollen in the patients.     

Presence ofAspergillus Mich. ex Fr. in several different biotopes in Córdoba (Spain)

The medical, veterinary and saprophytic importance ofAspergillus has been acknowledged by many authors. This paper reports on the occurrence ofAspergillus species in the air and dust of Córdoba (Spain). Four studied were done: first, on the air of three rooms and the outside area of 14 homes over 2 years; second, outdoor air in the west of the central district, with three samples daily over 1 year; third, the air inside and outside four of the largest silos and commercial grain warehouses in the province, over 1 year: and fourth, the dust of 12 primary schools in Córdoba city, for two consecutive school years. For air-sampling, volumetric and gravimetric methods were used, both with a culture medium and with 20-min exposure. For dust-sampling, a customised vacuum cleaner was used; samples collected were inoculated onto Petri dishes containing culture medium. A total of 38Aspergillus species were identified (we also detected four ascosporic states). A comparison of the results obtained at each of the sampling sites was made and an analysis of variance revealed quantitatively significant differences between sampling times.     

Effect of tert-butyl hydroperoxide addition on spontaneous chemiluminescence in brain

It is well known that light emission is related to lipid peroxidation in biological material, and that this process occurs spontaneously in the brain. tert-Butyl hydroperoxide (tBHP) is an organic peroxide widely used as initiator of free radical production in several biological systems. However, the prooxidant capacity of this compound remains unclear. To clarify its role in brain spontaneous autooxidation, rat brain homogenates were incubated with and without tBHP. Light emission and lipid peroxidation were measured by luminometry and the TBARs test, respectively. Several inhibitors of free radical-induced lipid peroxidation were also used. These inhibitors included ascorbate, EDTA, and desferrioxamine. Our results indicate that the pattern of light emission spontanously produced in brain was different from that observed after the addition of tBHP to the homogenates, and that these differences depended on the tBHP concentration. The main difference was that tBHP caused a rapid light emission that reached its maximum more quickly than in the case of spontaneous emission. Addition of ascorbate resulted in an increase in chemiluminescence in presence of tBHP. In contrast, EDTA and desferrioxamine inhibited light emission in homogenates both with and without tBHP. The results of MDA determination were similar to those described, including the effect of inhibitors. A common feature in MDA and luminometric determinations was the dispersion of data. In conclusion, these results suggest that tBHP, under specific conditions, modify the kinetic pattern of brain spontaneous autooxidation.     

Die centro-epigenetische Hypothese und der Einfluß des Centralnervensystems auf embryonale Entwicklung und Regeneration

Ohne Zusammenfassung     

Regiospecificity of Two Multicomponent Monooxygenases from Pseudomonas stutzeri OX1: Molecular Basis for Catabolic Adaptation of This Microorganism to Methylated Aromatic Compounds

The pathways for degradation of aromatic hydrocarbons are constantly modified by a variety of genetic mechanisms. Genetic studies carried out with Pseudomonas stutzeri OX1 suggested that the tou operon coding for toluene o-xylene monooxygenase (ToMO) was recently recruited into a preexisting pathway that already possessed the ph operon coding for phenol hydroxylase (PH). This apparently resulted in a redundancy of enzymatic activities, because both enzymes are able to hydroxylate (methyl)benzenes to (methyl)catechols via the intermediate production of (methyl)phenols. We investigated the kinetics and regioselectivity of toluene and o-xylene oxidation using Escherichia coli cells expressing ToMO and PH complexes. Our data indicate that in the recombinant system the enzymes act sequentially and that their catalytic efficiency and regioselectivity optimize the degradation of toluene and o-xylene, both of which are growth substrates. The main product of toluene oxidation by ToMO is p-cresol, the best substrate for PH, which catalyzes its transformation to 4-methylcatechol. The sequential action of the two enzymes on o-xylene leads, via the intermediate 3,4-dimethylphenol, to the exclusive production of 3,4-dimethylcatechol, the only dimethylcatechol isomer that can serve as a carbon and energy source after further metabolic processing. Moreover, our data strongly support a metabolic explanation for the acquisition of the ToMO operon by P. stutzeri OX1. It is possible that using the two enzymes in a concerted fashion confers on the strain a selective advantage based on the ability of the microorganism to optimize the efficiency of the use of nonhydroxylated aromatic hydrocarbons, such as benzene, toluene, and o-xylene.     

Amos Arthur Heller’s Puerto Rico plant collecting itineraries of 1900 and 1902–1903 and their utility for the historical study of endangered plants

This work reports the Puerto Rico plant collecting itineraries of 1900 and 1902–1903 of Amos Arthur Heller based on the original notebooks recently located at the Herbarium of the University of Washington-Seattle. The utility of historical data for understanding original distributions of rare species is demonstrated for two Puerto Rican species:Buxus vahlii Ball andDaphnopsis helleriana Urb. Urb.     

Massive corals in Paleocene siliciclastic sediments of Chubut (Argentina)

A horizon with large, massive corals in growth position was discovered in the Paleocene, probably upper Danian, part of the Maastrichtian–Paleocene Lefipán Formation of Chubut (Patagonia, Argentina). All corals belong to one species, the cosmopolitan Haimesiastraea conferta Vaughan, which survived the end-Cretaceous mass extinction. The occurrence of massive corals at this site is exceptional both because of the siliciclastic depositional regime and because of the high palaeolatitude setting. An unusual autecology of this coral and strongly reduced sedimentation rates, were probably the prerequisites for coral growth, but a link to palaeoclimate is less likely.     

Fhit interaction with ferredoxin reductase triggers generation of reactive oxygen species and apoptosis of cancer cells

Fhit protein is lost in most cancers, its restoration suppresses tumorigenicity, and virus-mediated FHIT gene therapy induces apoptosis and suppresses tumors in preclinical models. We have used protein cross-linking and proteomics methods to characterize a Fhit protein complex involved in triggering Fhit-mediated apoptosis. The complex includes Hsp60 and Hsp10 that mediate Fhit stability and may affect import into mitochondria, where it interacts with ferredoxin reductase, responsible for transferring electrons from NADPH to cytochrome P450 via ferredoxin. Viral-mediated Fhit restoration increases production of intracellular reactive oxygen species, followed by increased apoptosis of lung cancer cells under oxidative stress conditions; conversely, Fhit-negative cells escape apoptosis, carrying serious oxidative DNA damage that may contribute to an increased mutation rate. Characterization of Fhit interacting proteins has identified direct effectors of the Fhit-mediated apoptotic pathway that is lost in most cancers through loss of Fhit.     

Human-murine transthyretin heterotetramers are kinetically stable and non-amyloidogenic. A lesson in the generation of transgenic models of diseases involving oligomeric proteins

The transthyretin amyloidoses appear to be caused by rate-limiting tetramer dissociation and partial monomer unfolding of the human serum protein transthyretin, resulting in aggregation and extracellular deposition of amorphous aggregates and amyloid fibrils. Mice transgenic for few copies of amyloid-prone human transthyretin variants, including the aggressive L55P mutant, failed to develop deposits. Silencing the murine transthyretin gene in the presence of the L55P human gene resulted in enhanced tissue deposition. To test the hypothesis that the murine protein interacted with human transthyretin, preventing the dissociation and partial unfolding required for amyloidogenesis, we produced recombinant murine transthyretin and human/murine transthyretin heterotetramers and compared their structures and biophysical properties to recombinant human transthyretin. We found no significant differences between the crystal structures of murine and human homotetramers. Murine transthyretin is not amyloidogenic because the native homotetramer is kinetically stable under physiologic conditions and cannot dissociate into partially unfolded monomers, the misfolding and aggregation precursor. Heterotetramers composed of murine and human subunits are also kinetically stable. These observations explain the lack of transthyretin deposition in transgenics carrying a low copy number of human transthyretin genes. The incorporation of mouse subunits into tetramers otherwise composed of human amyloid-prone transthyretin subunits imposes kinetic stability, preventing dissociation and subsequent amyloidogenesis.