Decreased expression o ubiquinol-cytochrome c reductase subunits in patients exhibiting mitochondrial myopathy with progressive exercise intolerance

The expression of mitochondrial proteins of two patients suffering from myopathy with progressive exercise intolerance and exhibiting a deficiency in the enzymatic activity of complex III (ubiquinol-cytochrome c reductase) has been analyzed by immunological titration. In both patients, the Fe-S protein, the cytochrome b and the 9.5 kDa protein were decreased while the expression of the other complex III subunits were close to normal values. This data indicates that, in some mitochondrial myopathies, proteins of the respiratory chain complexes can be accumulated in mitochondria without being integrated into a functional complex. This may be explained either by a lack of control of the coordination between the synthesis of subunits of mitochondrial and nuclear origin or by a difference in the degradation rate of the various subunits which are not properly assembled.     

Efficient allelic exchange and transposon mutagenesis in Mycobacterium tuberculosis

A better understanding of Mycobacterium tuberculosis virulence mechanisms is highly dependent on the design of efficient mutagenesis systems. A system enabling the positive selection of insertional mutants having lost the delivery vector was developed. It uses ts-sacB vectors, which combine the counterselective properties of the sacB gene and a mycobacterial thermosensitive origin of replication and can therefore be efficiently counterselected on sucrose at 39°C. This methodology allowed the construction of M. tuberculosis transposition mutant libraries. Greater than 10⁶ mutants were obtained, far exceeding the number theoretically required to obtain at least one insertion in every nonessential gene. This system is also efficient for gene exchange mutagenesis as demonstrated with the purC gene: 100% of the selected clones were allelic exchange mutants. Therefore, a single, simple methodology has enabled us to develop powerful mutagenesis systems, the lack of which was a major obstacle to the genetic characterization of M. tuberculosis.     

Liver and colon DNA oxidative damage and gene expression profiles of rats fed Arabidopsis thaliana mutant seeds containing contrasted flavonoids.

Plant polyphenols, such as flavonoids, comprise many compounds, ranging from simple phenolic molecules (i.e. flavonols, anthocyanins) to polymeric structures with high molecular weight (as proanthocyanidins, PAs). We investigated the effects of flavonoids by feeding Wistar rats Arabidopsis thaliana seeds carrying mutations in key enzymes of the flavonoid biosynthetic pathway (15% w/w seeds for 4 weeks). The seeds used were: Ws-2 wild-type containing flavonols and PAs, tt3-4 mutant containing flavonols only, ban-5 accumulating flavonols and anthocyanins, tt4-8 mutant, deprived of flavonoids. DNA oxidative damage was significantly reduced only in the liver of rats fed tt3-4 mutant seeds. Microarray analysis of the liver revealed down-regulation of genes associated with oxidative stress, Krebs cycle, electron transport and proteasome degradation in all experimental groups compared to the tt4-8-fed reference rats; therefore, these effects were due to the flavonol content and not to high molecular weight compounds. We observed a down-regulation of inflammatory response genes in the colon mucosa in ban-5- fed rats, probably due to anthocyanin content. In conclusion, flavonols exhibited antioxidant effects at systemic level, whereas high molecular weight flavonoids affected only the colon, probably due to their limited absorption.     

Characteristics of the cholera epidemic of 1998 in Ecuador during El Ni?o]

OBJECTIVE: To describe the outbreak of cholera that occurred in Ecuador in 1998 during the El Nino weather phenomenon, to present data on the resistance of the circulating strains of Vibrio cholerae to antimicrobial drugs, and to describe the preventive measures taken by health authorities in order to reduce the impact of the disease. METHODS: The epidemiological data came from three sources: 1) the registry of the National Bureau of Epidemiology of the Ministry of Public Health of Ecuador, 2) the registry of the National Institute of Hygiene and Tropical Medicine, and 3) the final report of the Training Program for the Fight against Cholera and Diarrheal Diseases. Isolation, identification, and serotyping was done of V. cholerae in the feces samples from 10% of the suspected cholera cases that were identified between 1 January and 31 December 1998. The suspected cases were defined by the sudden appearance of watery diarrhea, with or without dehydration, in epidemic areas. The strains that were isolated were submitted to a standard antibiogram by the diffusion method, in which the following antibiotics were tested: amoxicillin, tetracycline, trimethoprimsulfamethoxazole, vibriostatic compound O/129, nalidixic acid, erythromycin, norfloxacin, ciprofloxacin, gentamycin, chloramphenicol, and colistin. RESULTS: In 1998 there were 3 755 cases reported in 17 of the 21 provinces of the country. This corresponds to an incidence rate of 53.96 per 100 000 population. Thirty seven patients died, for a case fatality rate of 0.97%. A total of 301 strains of V. cholerae were isolated in the 637 suspected-cholera samples that were processed; all corresponded to V. cholerae O1, El Tor, subtype Ogawa. All of the strains were sensitive to tetracycline and to quinolones; 5.6% of the strains were resistant to erythromycin. The only strain resistant to amoxicillin was multiresistant. Officials in Ecuador implemented a series of preventive measures, and the surveillance system was strengthened in order to reduce the impact of the disease. CONCLUSIONS: The preventive measures helped to reduce the impact of the 1998 cholera epidemic in Ecuador, in terms of both incidence and the case fatality rate. Given the overall sensitivity of the strains to the antimicrobial drugs, there is no reason to change the current treatment regimens in the country. Taking into account the frequency of natural disasters in Ecuador and the relation that they have to the reappearance of cholera, interventions should be designed that make it possible to prevent and control the reappearance of the disease and its spread to the most vulnerable provinces of the central Sierra mountainous region and the eastern part of the country.     

A cysteine-rich motif confers hypoxia sensitivity to mammalian large conductance voltage- and Ca-activated K (BK) channel alpha-subunits

Cellular responses to hypoxia are tissue-specific and dynamic. However, the mechanisms that underlie this differential sensitivity to hypoxia are unknown. Large conductance voltage- and Ca-activated K (BK) channels are important mediators of hypoxia responses in many systems. Although BK channels are ubiquitously expressed, alternative pre-mRNA splicing of the single gene encoding their pore-forming alpha-subunits provides a powerful mechanism for generating functional diversity. Here, we demonstrate that the hypoxia sensitivity of BK channel alpha-subunits is splice-variant-specific. Sensitivity to hypoxia is conferred by a highly conserved motif within an alternatively spliced cysteine-rich insert, the stress-regulated exon (STREX), within the intracellular C terminus of the channel. Hypoxic inhibition of the STREX variant is Ca-sensitive and reversible, and it rapidly follows the change in oxygen tension by means of a mechanism that is independent of redox or CO regulation. Hypoxia sensitivity was abolished by mutation of the serine (S24) residue within the STREX insert. Because STREX splice-variant expression is tissue-specific and dynamically controlled, alternative splicing of BK channels provides a mechanism to control the plasticity of cellular responses to hypoxia.     

Allergy-like reactions to iodinated contrast agents. A critical analysis

Allergy-like reactions may occur following administration of iodinated contrast media (CM), mostly in at-risk patients (patients with history of previous reaction, history of allergy, co-treated with interleukin-2 or beta-blockers, etc.) but remain generally unpredictable. Severe and fatal reactions are very rare events. All categories of CM may induce such reactions, although first generation (high osmolar CM) have been found to induce a higher rate of adverse events than low osmolar CM. However, no differences were found between the two categories of CM with respect to mortality. Delayed reactions can also occur. There are no differences between the various categories of CM except for non-ionic dimers, which are more likely to induce such effect. Numerous clinical studies have evaluated the prophylactic value of drugs (mostly antihistamines and corticosteroids). Results are unclear and highly variable. Any prevention depends upon the mechanism involved. However, the mechanism of CM-induced allergy-like reaction remains disputed. Relatively recent data revived the hypothesis of a type-I hypersensitivity mechanism. Positive skin tests to CM have been reported. However, the affinity of IgE towards CM has been found to be very low in the only study which actually evaluated it. Other pathophysiological mechanisms (involving direct secretory effects on mast cells or basophils, or activation of the complement system associated or not with the plasma contact system) are also much debated. Anaphylaxis and anaphylactoid reactions are, in the end, clinically undistinguishable.     

Is fetal gender a risk factor for severe congenital cytomegalovirus infection?

Cytomegalovirus is the main cause of congenital viral infection and amniotic fluid viral load appears to be the single nonclinical prognostic factor. However, as in other infectious diseases, host genetics may influence the severity of the disease. To test this hypothesis, we looked retrospectively at the fetal gender in cases of severe congenital cytomegalovirus infection in our database. We also analyzed the international English literature covering this subject between 1985 and 2003. The proportion of females with brain abnormalities was statistically different from that of males (62/258: 24% vs 30/251: 12%, p = 0.004). The risk of abnormal brain development in infected fetuses was twice as high in females than in males (Chi(2) = 8.7; OR = 2, IC [1.26-3.21]). In our cases, amniotic fluid CMV DNA load was not significantly higher in males than in females (p = 0.06) and was also similar in severely and non-severely infected fetuses (p = 0.09).