Influence of culture conditions on the technological properties of Carnobacterium maltaromaticum CNCM I-3298 starters.

AIM: The aim of this study is to investigate the effect of a broad spectrum of culture conditions on the acidification activity and viability of Carnobacterium maltaromaticum CNCM I-3298, the main technological properties that determine the shelf-life of biological time-temperature integrator (TTI) labels. METHODS AND RESULTS: Cells were cultivated at different temperatures (20-37°C) and pH (6-9·5) according to a modified central composite design and harvested at increasing times up to 10 h of stationary phase. Acidification activity and viability of freeze-thawed concentrates were assessed in medium mimicking the biological label. Acidification activity was influenced by all three culture conditions, but pH and harvest time were the most influential. Viability was not significantly affected by the tested range of culture conditions. CONCLUSIONS: Carnobacterium maltaromaticum CNCM I-3298 must be cultivated at 20°C, pH 6 and harvested at the beginning of stationary phase to exhibit fastest acidification activities. However, if slower acidification activities are pursued, the recommended culture conditions are 30°C, pH 9·5 and a harvest time between 4-6 h of stationary phase. SIGNIFICANCE AND IMPACT OF THE STUDY: Quantifying the impact of fermentation temperature, pH and harvest time has led to a predictive model for the production of biological TTI covering a broad range of shelf-lives.

[Perforated jejunal tuberculosis in a patient with HIV-infection]. / Tuberculosis yeyunal perforada en paciente con infección por VIH.

Patients with AIDS are particularly susceptible to tuberculosis infection with a high incidence of extrapulmonary disease and surgical complications. Authors describe a 38-year-old male infected with the human immunodeficiency virus who presented intestinal perforation due to mycobacterium tuberculosis. A resection of jejunum was performed with primary anastomosis. The postoperative course was further compromised by hepatic failure and the patient died 16 days after the initial surgery.

[Mortality related to alcohol consumption in Spain: 1981-1990]. / Mortalidad relacionada con el consumo de alcohol en España: 1981-1990.

OBJECTIVE: The aim of this study was to assess the overall contribution of alcohol to Spanish mortality during 1981 to 1990, as well as the impact on the premature death. METHODS: To this purpose we have used the sources of data furnished by the 'Movimiento Natural de la Población' that provides data of causes of death. Figures of proportional mortality, adjusted mortality and years of potential life lost were calculated, as well as trend analysis. RESULTS: 6.3% (mean in the ten years period) of the mortality was due to alcohol. This mortality was higher among males than females. Adjusted mortality show a light increase during the period. The most important category referring to years of potential life lost was unintentional injuries. In this category, motor vehicle accidents were responsible for the majority of premature death. CONCLUSION: This study shows the importance of alcohol related mortality in our country and the large premature death.

[Painful erections related to sleeping]. / Erecciones dolorosas relacionadas con el sueño.

Case report of sleep-related painful erections in a 34 year-old male with grade C3 HIV infection. Due to severe impairment of the patient's general condition, no proper diagnostic studies were performed to gain deeper knowledge of the symptom's pathological etiology. Empirical therapy was started based on evidence from the literature consulted, and the results seen were optimal. This paper contributes a brief review of a condition infrequently seen by the vast majority of urologists.

[Chorioretinitis caused by toxoplasmosis: study of 65 cases]. / Coriorretinitis por toxoplasmosis: estudio de sesenta y cinco casos.

Sixty five patients with an ophthalmoscopic diagnosis of chorioretinitis who, underwent complement fixation tests and intradermoreaction using Toxoplasma antigens were studied. 95,4% of them disclosed antibody titers with the former and 58,5% reactors were found with the latter. Sixty five individuals with neither backgrounds nor clinical symptoms of toxoplasmosis were used as control, and 27,7% of reactors in both tests were likewise found. The differences among results are highly statistically significant; this points out toxoplasmosis as an important cause of chorioretinitis in our patients. Complement fixation tests were more sensitive than intradermoreactions as well as specific as the latter. Results were correlated to age, sex and animal contacts.

Permeability to alpha sarcin in virus-infected cells.

Incubation of HeLa cells with Encephalomyocarditis virus (EMC) induces permeability of the cell membrane to protein toxins, such as alpha sarcin. To induce permeability to this toxin only 5 min incubation of cells with virus is needed. On the other hand, less than 1 min exposure of the susceptible cells to alpha sarcin produces maximal inhibition of protein synthesis. EMC virus treated with UV-light, although unable to replicate, can still induce the entrance of alpha sarcin into HeLa cells, but the virion loses this capacity after heating at 60 degrees C for 10 min. These findings suggest that an integral viral genome is not necessary to make the cells permeable to alpha sarcin, and that a virion protein might be involved in this phenomenon. Although human interferon prevents productive EMC infection, it does not affect the virus-induced entrance of alpha sarcin into the cells. The plasma membrane of cells that have been treated with virion particles can recover its initial lack of permeability to alpha sarcin after 2 h at 37 degrees C. Poliovirus modifies membrane permeability in human HeLa cells, but it has no effect on mouse L cells. This fact suggests that viral attachment to specific cell surface receptors is necessary to induce permeability, since receptors to poliovirus are only present in primate cells.

Permeability to inhibitors of protein synthesis in virus infected cells.

Infection of HeLa cells with different viruses induces permeabilization of the cell membrane to protein toxins such as alpha-sarcin. This phenomenon occurs with HeLa, KB, BHK-21 and L929 cells and EMC, SFV, VSV and Polio virus and is dependent on the ability of the virus to infect the cells. Inhibitors of endocytosis and lysosomotropic agents do not affect this process. Cells become sealed to the toxin approximately four hours after the infection. Sulfhydryl reagents impair cellular permeabilization to alpha-sarcin.

Dissociative flashbacks after right frontal injury in a vietnam veteran with combat-related posttraumatic stress disorder.

A Vietnam veteran with a combat-related posttraumatic stress disorder developed recurrent dissociative flashbacks (related to the atrocities of a specific war incident) several months after suffering a traumatic brain injury. CT disclosed a small lesion in the right dorsolateral prefrontal cortex. SPECT demonstrated more extensive functional changes in prefrontal and anterior paralimbic brain regions, mainly in the right hemisphere. This case further implicates the provocative effect of physical stimuli (brain damage) in reawakening old dormant memories and the preferential role of the right hemisphere for the storage of traumatic memories.

Effects of ricin on the ribosomal sites involved in the interaction of the elongation factors.

The effects of ricin on the different steps of the elongation cycle of protein synthesis in a rabbit reticulocyte cell-free system are studied in this paper. The toxin most probably acts by catalytically inactivating the ribosomes, since a single molecule of the toxin can inactivate 300 ribosomes for poly(U)-directed phenylalanine incorporation. The effect of the toxin on the ribosome is irreversible. Ricin specifically inhibits elongation-factor-1-dependent aminoacyl-tRNA binding to ribosomes but has no effect on the non-enzymic binding of aminoacyl-tRNA. Ricin also inhibits formation of the complex elongation-factor-2 - ribosome - nucleotide with GTP, GDP or GMP-P(CH2)P. However, the toxin has no effect on translocation. These apparently conflicting results are discussed in this study.

Antibiotics and compounds affecting tanslation by eukaryotic ribosomes. Specific enhancement of aminoacyl-tRNA binding by methylaxnthines.

The mode and site of action of inhibitors of translation (initiation, elongation and termination of protein synthesis) in eukaryotic systems is reviewed. The isolation and characterization of a factor is described that binds Ac-Phe-tRNA to form a complex made up of binding factors, Ac-Phe-tRNA, and ribosome. The binding of Ac-Phe-tRNA probably occurs at the ribosomal site involved in the binding of the initiator substrate Met-tRNAF. The effect of inhibitors of the intitiation phase of protein synthesis on the nonenzymic Ac-Phe-tRNA binding to ribosomes is investigated. The two sites translocation model for translation in eukaryotic cells is presented and the effects of inhibitors on the various steps of protein synthesis are determined empirically. The site of action of inhibitors of peptide bond formation at the ribosomal peptidyl transferase center is elucidated. The action of inhibitors of translocation is sutdied in model cell-free systems from human cells. In addition, a number of methylxanthines are shown to enhance the elongation phase in polypeptide synthesis by stimulating the enzymic binding of aminoacyl-tRNA. The effect of caffeine, theophylline and its derivatives are shown to be fairly specific and dependent on the ribosome concentration. Aminophylline is shown to have a similar effect but also enhances aminoacyl-tRNA synthetase activity at low Mg++ concentrations, probably displacing the optimal concentration of Mg++ in the reaction. This second effect of aminophylline appears to be due to the ethylenediamine moiety of aminophylline since it is also observed in the presence of different polyamines but not in the presence of caffeine or theophylline.

Protective effect of elongation factor 2 on the inactivation of ribosomes by the toxic lectins abrin and ricin.

The inactivation of rabbit reticulocyte ribosomes by abrin and ricin A-chains was studied by incubating ribosomes with the A-chains and testing, after various periods of time, aliquots of the ribosomes for their ability to polymerize phenylalanine. The presence of elongation factor 2 (EF-2) reduced the rate of inactivation of ribosomes by the A-chains. The protective effect of EF-2 was strongly enhanced by GTP and, to a lesser extent, also by GDP or dGTP. Other nucleotides had no demonstrable effect. Much less protection was found after binding of Phe-tRNA to ribosomes in the presence of EF-1 (enzymic binding) or in the presence of high Mg2+ concentration (non-enzymic binding). The data indicate that when EF-2 binds to the ribosomes it completely or partially covers the target site for abrin and ricin A-chains. The possibility that EF-1 also binds to this site is discussed.

Ribosome inactivation by the toxic lectins abrin and ricin. Kinetics of the enzymic activity of the toxin A-chains.

A sensitive test system for toxin-treated ribosomes was worked out by treating rabbit reticulocyte ribosomes with abrin A-chain, ricin A-chain or ricinus agglutinin A-chain, adding neutralizing amounts of specific antitoxins and testing for polyphenylalanine-synthesizing activity in a system where the concentration of elongation factors and ribosomes were varied. The strongest inhibition was obtained in the presence of low concentrations of elongation factor (EF-2). The activity of the ribosomes decreased with time of incubation with the toxin A-chains. Addition of anti-toxins stopped further inactivation. In systems containing untreated and toxin-treated ribosomes the ability to polymerize phenylalanine was proportional to the concentration of untreated ribosomes. There was a linear relationship between toxin A-chain concentration and the number of ribosomes inactivated per minute. The inactivation rate increased with temperature, and the estimated activation energy was 10.6 kcal (44.3 kJ). Linewaver-Burk plots of the data obtained by incubating various ribosome concentrations with toxins indicated a molecular activity of about 1500 ribosomes/minute for abrin and ricin A-chains and 100 ribosomes/minute for ricinus agglutinin A-chain. The apparent Michaelis constant was 0.1-0.2 muM for all three A-chains. The activity of the A-chains in the intact cell is discussed.

Site of action of ricin on the ribosome.

The extent of the inhibitory effect of ricin in polyphenylalanine synthesis by eukaryotic ribosomes is strongly dependent upon the concentration of ribosomes and the elongation factors EF 1 and EF2. Maximal inhibition by ricin, in this assay is observed when either ribosomes or the two elongation factors are added under limiting conditions, whereas ricin-treated ribosomes support protein synthesis at saturating concentrations of elongation factors and ribosomes. Similarly, the enzymatic binding of Phe-tRNA to ribosomes is drastically blocked in ricin-treated ribosomes when low EF 1 concentrations are added to the reaction mixture, but there is no inhibition when EF 1 is at saturating concentrations. Furthermore, formation of the complex EF 2-guanosine triphosphate-ribosome, using free ribosomes pretreated with ricin, is strongly inhibited at limiting concentrations of EF2, but is not affected at saturating concentrations of this factor. However, ricin does not inhibit the EF 2-dependent translocation of peptidyl-tRNA by polysomes, although the toxin is very active in preventing amino acid incorporation by polysomes. Our results suggest that the damaging effect of ricin on the ribosome causes a decreased affinity for both elongation factors EF 1 and EF 2. Thus, the toxin inhibits the enzymatic binding of aminoacyl-tRNA to ribosomes. The lack of inhibition of translocation by ricin suggests that the toxin cannot interact with ribosomes with substrate bound to the acceptor site. Essentially similar results are observed with ricin, abrin, ricin A chain, abrin A chain, and ricinus agglutinin A chain. A possible effect of the toxins on initiation and/or termination is further discussed.