Community-acquired pneumonia due to Acinetobacter lwoffii in a patient infected with the human immunodeficiency virus.
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Biomedical subjects
Publications and source records attributed to R Muñoz.
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We report the cloning of the gyrB gene from Streptococcus pneumoniae 533 that carries the nov-1 allele. The gyrB gene codes for a protein homologous to the gyrase B subunit of archaebacteria and eubacteria. The same amino acid substitution (Ser-127 to Leu) confers novobiocin resistance on four isolates of S. pneumoniae. This amino acid position is equivalent to Val-120 of Escherichia coli GyrB, a residue that lies inside the ATP-binding domain as revealed by the crystal structure of the protein.
Melon (Cucumis melo L.) dry seeds contain melonin, a protein that strongly inhibits ribosomes from different prokaryotic and eukaryotic sources including those from melon. The protein was purified by a new method to yield highly active and stable protein preparations that involves chromatography through S-Sepharose Fast Flow, CM-Sepharose, Superdex 75 and Mono-S. Melonin shows important functional properties: 1) its inhibitory effects on translation were irreversible; 2) it is a single unglycosylated polypeptide chain with an apparent M(r) of 22000; 3) it degrades RNA in a dose-dependent way without affecting DNA. In the light of present results melonin can be considered as a new plant RNase of unusual properties.
Dry seeds of Cucumis sativus L. were found to contain a heat-sensitive endoribonuclease of a novel type which we have named cusativin. It was purified to apparent electrophoretic homogeneity by chromatography through S-Sepharose Fast Flow, Sephadex G-75, CM-Sepharose, Superdex 75-FPLC (fast protein liquid chromatography) and Mono S-FPLC. It is a single unglycosylated polypeptide chain with an apparent molecular mass (M(r)) of 22900. Polyclonal anti-cusativin antibodies raised in rabbits only reacted with melonin, the translation inhibitor from Cucumis melo L. Functional, Western blot and enzyme-linked immunosorbent assay (ELISA) analyses indicated that cusativin is present in the coat and cotyledons of dry seeds, but not in embryonic axes. Cusativin is accumulated in maturing seeds. By contrast, after seed germination there is degradation of the cusativin present in cotyledons but not that present in the seed coat. The preference of cusativin for polynucleotide cleavage was poly(C) >> poly(A) acids, poly(U) and poly(G) being unaffected by cusativin. Under the denaturing conditions used for RNA sequencing, cusativin acted only on poly(C). Cusativin proved to be useful for RNA sequencing, in particular, complementing the data obtained with RNase CL3. Cusativin represents a new class of plant RNase and, as far as we are aware, is the first plant enzyme that shows cleavage specificity for cytidine under the denaturing conditions of RNA sequencing.
Two new N-glycosidase type-1 ribosome-inactivating proteins (RIPs), denoted petroglaucin 1 and petrograndin, respectively, were isolated from the plants Petrocoptis glaucifolia (Lag.) Boiss sp. viscosa (Rothm.) Lainz and Petrocoptis grandiflora Rothm. These new RIPs do not share H2N-terminal amino-acid sequence homology with petroglaucin (now denoted as petroglaucin 2), the only other type-1 RIP to be isolated from P. glaucifolia (Arias et al. (1992) Planta 186, 532-540). Petroglaucin 1 shares amino-acid sequence homology with RIPs from Cucurbitaceae while petroglaucin 2 and petrograndin do so with saporins and dianthin 30 (Caryophyllaceae). The new RIPs strongly inhibited protein synthesis at subnanomolar concentrations in rabbit reticulocyte lysates and other eukaryotic cell-free systems, but they were inactive on bacterial ribosomes.
The oxidation of hydroquinone by two peroxidase (EC 1.11.1.7) fractions obtained from the cells and spent medium of cell cultures of grapevine (Vitis vinifera cv Monastrell) has been studied, and their comparative efficacy (kcat/KM ratio) studied in both the H2O2-consuming and hydroquinone-consuming reactions. While the efficacy in the H2O2-consuming reaction is practically identical for both enzyme fractions, the cellular peroxidase has five-fold more efficacy in the hydroquinone-consuming reaction than the peroxidase located in the spent medium. Screening of cellular peroxidases capable of oxidizing hydroquinone on polyacrylamide gels, by means of a staining reaction based on the nucleophilic attack of 4-aminoantipyrine on p-benzoquinone in acidic media, reveals that all the cellular peroxidase isoenzymes are capable of oxidizing hydroquinone, probably yielding a quinone-diimine as a product of the staining reaction. Since isoperoxidases found in cellular fractions are also present in the spent medium, the values found for the different efficacies in the hydroquinone-consuming reaction must be considered as the results of the different proportions in which each peroxidase isoenzyme was found in the two fractions. The localization of a benzoquinone-generating system of high efficacy inside the plant cell, and probably located in vacuoles, is discussed with respect to the harmful role which the quinone/semiquinone pair might play in cell death, as part of the hypersensitive response expressed within the mechanism of plant disease resistance.
Resistance to third-generation cephalosporins in a clinical isolate of Streptococcus pneumoniae was shown to be due to the production of altered forms of penicillin-binding proteins (PBPs) 2X and 1A. The cloned PBP2X gene from the resistant strain was able to transform a susceptible strain to an intermediate level of resistance. The resulting transformant could be transformed to the full level of resistance of the clinical isolate using the cloned PBP1A gene from the latter strain. Chromosomal DNA from the resistant strain (and from other resistant strains) could readily transform a susceptible strain to the full level of resistance to third-generation cephalosporins (greater than 250-fold for cefotaxime; greater than 100-fold for ceftriaxone) in a single step (transformation frequency of about 10(-5)). The resistant transformants obtained with chromosomal DNA were shown by gene fingerprinting to have gained both the PBP1A and PBP2X genes from the DNA donor.
The inhibitory effect of indole-3-acetic acid, and of its peroxidase-mediated degradation products of an indole nature, on the oxidation rate of coniferyl alcohol catalyzed by cell wall peroxidases has been studied. The results show that the inhibitory effect of indole-3-acetic acid and indole-3-carbinol may be explained, in part, by their properties as peroxidase substrates. However, I50 values for a series of indole compounds not regarded as peroxidase substrates show a good correlation with the electron-donating or electron-withdrawing nature of the 3-substituents, as judged by the linearity of the Hammett rho sigma plot. These results suggest that although the properties of indole compounds as peroxidase substrates may be responsible, in part, for their inhibitory effects on the peroxidase-mediated oxidation of coniferyl alcohol, the inhibitory effect appears to be mainly determined by the acidity of the imino group of the indole nucleus.
The present screening work was devoted to the search for new ribosome-inactivating proteins (RIPs) in 21 plant species. Eight plants proved to be very active, inhibiting protein synthesis in eukaryotic in vitro systems (rat liver, Vicia sativa and wheat germ). They fulfil the major requirements for consideration as type 1 RIPs. Also, eight plants were found to contain haemagglutinating activity of human red cells but this was not related to the simultaneous presence of RIPs.
An approximation of Parkinson's disease in Spain was carried out based on the data of mortality by this disease over the period 1980-1985. The annual number of deaths by Parkinson's disease as well as its distribution by sex, age groups and Spanish provinces was obtained from information published annually by the National Institute of Statistics in the Natural Movement of the Spanish Population. The global mortality rate during the period studied was of 2.14 per 100,000 inhabitants. The specific mortality rate by Parkinson's disease in males was slightly higher than that of females. A specific mortality rate was also observed by age groups being higher in the older age groups. From the point of geographical distribution, higher rates were observed in the northern provinces with respect to the south and in rural areas compared to provinces capitals. Following the analysis of the results obtained and upon comparing the findings published in other countries it was concluded that place of residence may be related to the development of Parkinson's disease. From these data new epidemiological studies are required oriented to the identification of the environmental factors which may play a role in the etiology of this entity.
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Addition of the L-proline analogue L-azetidine 2-carboxylic acid to growing cultures of Saccharomyces cerevisiae var. ellipsoideus promoted fast deactivation of the general aminoacid permease, measured as L-valine uptake, without an immediate decrease in the growth rate. Cells preincubated with the analogue for 3 h were unable to restore either growth ability or general aminoacid permease activity in analogue-free medium. Eadie-Hofstee plots of L-valine uptake in the presence of the analogue are consistent with a strong reduction in the number of active molecules of the general amino-acid permease located in the plasma membrane. Inhibitory effects on protein synthesis were seen after preincubations of the yeast with the analogue for 3 h although a 30 min preincubation had no effect.
Long-term treatment of rats with isopropanol in the drinking water results in a change or process of adaptation occurring in in vitro brain protein synthesis which increases the resistance of the ribosomal machinery to the acute effect of either ethanol or isopropanol. Such an increase was observed both in the system coded by endogenous messenger and in the system coded by polyuridylic acid. In both translation systems, the adaptation seems to affect the ribosomal step of polypeptide chain elongation. The increase in resistance to the alkanols apparently did not affect the inhibitory action of puromycin, fusidic acid and cycloheximide on the ribosome.
Isolates of serotype 23F Streptococcus pneumoniae with high levels of resistance of penicillin have been commonly recovered in Spain for more than a decade. Recently penicillin-resistant serotype 23F S. pneumoniae strains were also isolated from children attending a day-care center in Cleveland. A number of Spanish and Cleveland isolates were compared by electrophoretic analysis of penicillin-binding protein (PBP) profiles and DNA restriction endonuclease cleavage profiles of the PBP 2X and 2B genes amplified with the polymerase chain reaction and by multilocus enzyme electrophoresis. All strains were identical by these criteria. The findings demonstrate that the Spanish and Cleveland isolates are clonally related and suggest that this antibiotic resistant clone of serotype 23F S. pneumoniae has spread intercontinentally from Spain to the United States.
For several years now, it has been known that the administering of adrenergic beta antagonists, especially of the beta-2 type, induce hypokalemia as a result of the entering of potassium into the skeletal muscle cells. This fall in kalemia occurs independently from the effect of insulin, aldosterone or kidney excretion, is mediated by the beta-2 receptors and require the intervention of cAMP joined at the cell membrane and the subsequent stimulation of the Na-K-ATPase which bring the potassium into the striated muscle cell. Among the most outstanding drugs with beta-2 effect is salbutamol, which maintains the hypokalemic effect whether administered intravenously or inhaled. It has been used in cases of hyperkalemia, in both children and adults. The initially used intravenous dosage (0.5 mg) caused several side-effects, especially rapid heart beat, seen more in children. It has been recently found that the use of doses as low as 4 micrograms/kg lower the kalemia to values averaging 1.4 to 1.6 mEq/L (mmol/L); in addition, using these dosages intravenously in an average of 20 minutes, no side-effects were seen, even when administered to newborns. For the above, we considered that salbutamol, in the suggested dosages, constitutes an efficient and secure therapeutic method for the initial treatment of severe hyperkalemic patients.
Long-term ingestion of sublethal n-butanol doses by rats led to a noteworthy increase in the resistance of in vitro brain ribosomal function to the acute inhibitory action of ethanol and isopropanol. Withdrawal of n-butanol did not change this adaptation process immediately. The step affected seems to be the elongation of polypeptide chains. The dependence of in vitro translation on incubation temperature was affected by the adaptation process, the translation system of chronic animals being less stimulatable than that of control animals at low temperature.
The oxidation of coniferyl alcohol (CA), a lignin precursor, by cell wall peroxidases may take place at the expense of indole-3-acetic acid (IAA) and O2, and in the absence of H2O2. The peroxidase-catalyzed oxidation of CA shows an optimum at an IAA concentration of 0.33 mM, while higher IAA concentrations are inhibitory. The observation that the oxidation of CA by cell wall peroxidase at the expense of IAA and O2 is inhibited by genistein, a putative endogenous inhibitor of lignification in lupin hypocotyls, supports the view that the H2O2-generating system coexists with cell wall peroxidase activities involved in lignification, and that it takes place at the expense of IAA and O2.
Long-term treatment of rats with n-butanol leads to a change in in vitro brain protein synthesis which increases the resistance of this process to either ethanol or isopropanol. The change seems to be related to ribosomal events since the synthesis of aminoacyl-tRNA was not affected in the same conditions.