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J Montoya

Publications and source records attributed to J Montoya.

At least 55 records · Page 3Linked to original sources
PubMed1996

Transient activation of mitochondrial translation regulates the expression of the mitochondrial genome during mammalian mitochondrial differentiation.

Regulation of the expression of the nuclear-encoded beta-subunit of H(+)-ATP synthase (beta-F1-ATPase) gene of oxidative phosphorylation during differentiation of liver mitochondria is mainly exerted at two post-transcriptional levels affecting both the half-life [Izquierdo, Ricart, Ostronoff, Egea and Cuezva (1995) J. Biol. Chem. 270, 10342-10350] and translational efficiency [Luis, Izquierdo, Ostronoff, Salinas, Santarén and Cuezva (1993) J. Biol. Chem. 268, 1868-1875] of the transcript. Herein, we have studied the expression of the mitochondrial (mt) genome during differentiation of rat liver mitochondria in an effort to elucidate the mechanisms of nucleo-mitochondrial cross-talk during biogenesis of the organelle. Estimation of the relative cellular representation of met-DNA in liver reveals a negligible increase in mt-DNA copy number during organelle differentiation. Concurrently, the lack of changes in transcription rates of the mt-DNA "in organello', as well as in steady-state levels of the mt-transcripts, suggests that organelle differentiation is not controlled by an increase in transcription of the mt-genome. However, translation rates in isolated mitochondria revealed a transient 2-fold increase immediately after birth. Interestingly, the transient activation of mitochondrial translation at this stage of liver development is dependent on the synthesis of proteins in cytoplasmic polyribosomes. These findings support the hypothesis that the expression of nuclear and mitochondrial genes during biogenesis of mammalian mitochondria is developmentally regulated by a post-transcriptional mechanism that involves concerted translational control of both genomes.

Aging↗
PubMed1996

The synthesis of mRNA in isolated mitochondria can be maintained for several hours and is inhibited by high levels of ATP.

The dependence for the maintenance of the synthesis and maturation of mitochondrial RNA on the supply of nucleo-cytoplasmic factors has been investigated by a novel in organello RNA synthesis system. We found that mitochondrial DNA transcription can be maintained for several hours in isolated mitochondria. Analysis of the individual mitochondrial RNA species revealed that: the processing of the rRNA precursors and the stability of the mature rRNAs, but not the transcription itself, is severely impaired after short periods of incubation, indicating that these processes are strongly dependent on the mitochondrial interaction with the nucleo-cytoplasmic compartment; the events that lead to the synthesis, processing and turnover of the mitochondrial mRNAs do not require the continuous supply of nucleo-cytoplasmic factors, that are accumulated in excess by mitochondria. Furthermore, we present evidence indicating an inhibition of high ATP levels on the mitochondrial RNA polymerase activity, both in organello and in vitro. Consequently, it is proposed that mitochondrial mRNA synthesis can be regulated in response to changes in intramitochondrial ATP levels. This regulation of mitochondrial mRNA synthesis together with their very rapid turnover described here and elsewhere [Gelfand, R. & Attardi, G. (1981) Mol. Cell Biol. 1, 497-511], could represent a mechanism that would allow each individual mitochondrion to adjust its optimal levels of mRNA, and hence its translation capacity, in response to local energetic demands.

Adenosine Triphosphate↗
PubMed1996

Bionomics of Lutzomyia evansi (Diptera: Psychodidae) vector of visceral leishmaniasis in northern Columbia.

The feeding behavior, seasonality, and natural infection rate of Lutzomyia evansi (Nuñez-Tovar) with Leishmania chagasi (Cuna & Chagas) was studied during a 12-mo period at 2 hamlets, El Contento and Vidales. Sand fly abundance in extra-, peri-, and intradomestic habitats was evaluated with sticky traps and CDC light traps, whereas human bait and Shannon trap collections were made only in peridomestic habitats. All trapping methods showed a clear predominance of L. evansi throughout the year. Sand flies were present during most of the year, with the exception of the driest months (February and March). Although the total number of sand flies was higher in El Contento than in Vidales, a larger proportion of L. evansi was found in intradomestic habitat than in the peri- and extradomestic habitats at Vidales. Also, sand flies from Vidales had a higher infection rate with L. chagasi than did those from El Contento. Although 2 of 9 promastigote infections detected in L. evansi were identified as L. chagasi, the difficulty of isolating and propagating leishmania strains from this visceral leishmaniasis focus precluded characterization of most parasite samples. Parous and infected sand flies were most abundant toward the end of the rainy season (October-December). For this reason, control strategies based on reducing sand fly populations or avoiding human-vector contact should be concentrated during the October-December period.

Animals↗
PubMed1995

mtDNA-depleted U937 cells are sensitive to TNF and Fas-mediated cytotoxicity.

It has been proposed that TNF cytotoxicity is mediated by reactive oxygen intermediates generated by uncoupling of mitochondrial respiration. We have compared sensitive U937 cells and derived cell lines depleted of mtDNA for their ability to undergo TNF- and Fas-induced apoptosis. Cells lacking around 98% of mtDNA were still sensitive to TNF-induced apoptosis. U937 cells devoid of mtDNA (U937-rho degree) were resistant to TNF, but this was due to the loss of its 55 kDa receptor. U937-rho degree cells were also resistant to docosahexaenoic acid, which causes U937 cell death by lipid peroxidation. These cells were sensitive to anti-Fas toxicity. The results indicate that TNF and Fas-induced toxicity occurs by a mechanism mostly independent of mitochondrial free radical generation.

Amobarbital↗
PubMed1995

[Progressive external ophthalmoplegia and the Kearns-Sayre syndrome: a clinical and molecular study of 6 cases].

The Kearns-Sayre syndrome (KSS) associates progressive external ophthalmoplegia initiating prior to the age of 20 years and pigmentary retinitis with a series of other heterogeneous clinical manifestations. The incomplete syndrome is usually denominated progressive external ophthalmoplegia (PEO)-plus which is a sporadically appearing mitochondrial cytopathy associated with large deletions of a variable proportion of mitochondrial DNA (mtDNA) molecules. Six patients with PEO-plus/KSS in whom muscle biopsy was performed following a complete clinical study are described. The muscle was processed by conventional histochemical techniques, electron microscopy, and genetic study (Southern transference, polymerase chain reaction, restriction cartography and both manual and automatic sequencing). The percentage of mutated mtDNA molecules for each patient was obtained by densitometry. The 6 patients presented multiorganic clinical manifestations characteristics of most mitochondrial diseases. The presence of destructured red fibers were observed in all the biopsies. All the patients presented a deletion in the mtDNA of a size between 4,861 to 7,437 base pairs (bp). All the deletions appeared flanked by direct repetitions from 4 to 13 bp and one also presented inverse repetitions from 5 to 6 bp in the zone next to the rupture point. In the 6 cases heteroplasmia was observed with a variable percentage of deleted molecules from 23 to 56%. The molecular basis of progressive external ophthalmoplegia-plus/Kearns-Sayre syndrome appears to be the existence of sole, large deletions in the mitochondrial DNA with the varying in location and percentage conditioning the appearance of different phenotypes similar among themselves. The 7,437 base pair deletion was the most frequently observed in the patients analyzed.

Adolescent↗
PubMed1995

Analysis of the transcription products of the rainbow trout (Oncorynchus mykiss) liver mitochondrial genome: detection of novel mitochondrial transcripts.

We have isolated and, by electrophoresis, using agarose slab gels in the presence of methylmercury hydroxide, analyzed the mitochondrial RNA content of the liver of rainbow trout. The RNAs corresponding to most of the mitochondrial DNA-encoded genes have been identified. Furthermore, among the transcription products we have also identified the nature of the RNA 8 previously described in human mitochondria, and detected a novel transcript that may represent the mRNA for ND6.

Animals↗
PubMed1995

Nucleotide sequence of the sheep mitochondrial DNA D-loop and its flanking tRNA genes.

The nucleotide sequence of the sheep mitochondrial DNA displacement-loop (D-loop) region and its flanking tRNA genes has been determined. Several conserved motifs among mammals have been identified along the 1189-bp sequence of the sheep control region: ten termination-associated sequences (TASs) and one conserved sequence block (CSB-1). CSB-2 and CSB-3, which are frequently found in most species, are not present in the sheep D-loop, which shows instead a short direct repeat at their usual localization. A long polypyrimidine tract between CSB-1 and the tRNA(Phe) gene is also present. Furthermore, the sheep mtDNA D-loop region displays tandem repeats in the left domain (adjacent to the tRNA(Pro) gene) comprising three different termination-associated sequences (TAS-5, TAS-6 and TAS-7).

Animals↗
PubMed1995

Phlebotomine sandflies associated with a focus of cutaneous leishmaniasis in Valle del Cauca, Colombia.

A survey was made of the phlebotomine sandfly fauna of La Guaira, a village with coffee plantations near Cali, Colombia, from which cases of American cutaneous leishmaniasis had been reported due to Leishmania (Viannia) panamensis and Le. (V.) braziliensis. Among six species of sandfly collected on human bait, Lutzomyia youngi was most important in terms of biting nuisance. Lu.columbiana, Lu.lichyi and Lu.scorzai as well as Lu.youngi adults occurred throughout the year. Sandfly man-biting activity occurred throughout the night and was highest within 2 h of sunset. Despite its abundance in nocturnal samples, Lu.youngi was rarely taken in diurnal resting site collections. In contrast, Lu.lichyi was collected on tree-trunks during the day in large numbers and was the only species biting in daylight. The implications of these and other findings for leishmaniasis control measures in La Guaira are considered.

Animals↗
PubMed1994

Highly efficient DNA synthesis in isolated mitochondria from rat liver.

We have developed a highly efficient DNA-synthesizing system with isolated intact rat liver mitochondria. The ATP requirements for this in organello DNA synthesis are provided by endogenous synthesis in the presence of exogenous ADP and an oxidizable substrate. In this system, mitochondrial DNA synthesis strikingly proceeds at a constant rate for about 5 h at 37 degrees C. Gel electrophoresis, hybridization and restriction enzyme analyses show that intact mitochondria synthesize nucleic acids with a size of 16.5 kb, that correspond to mitochondrial DNA, and that both DNA strands are replicated. This in organello DNA synthesis requires the supply of dNTPs and decreases at high ADP concentration in the incubation medium.

Adenosine Diphosphate↗
PubMed1994

Multiple deletions of mtDNA in two brothers with sideroblastic anemia and mitochondrial myopathy and in their asymptomatic mother.

Two brothers presented with a clinical picture characterized by sideroblastic anemia, mild pancreatic insufficiency and progressive muscle weakness. The presence of an associated permanent basal lactic acidemia raised the suspicion of a mitochondrial disease. A muscle biopsy performed in both siblings proved the presence of a significant number of ragged-red fibers, and respiratory chain enzymatic determinations demonstrated a reduced activity of complexes I, III and IV. Mitochondrial DNA studies disclosed the presence of multiple deletions both in skeletal muscle and, to a lesser extent, in leukocytes. Similar, but not identical deletions were also present in the leukocytes and muscle from their mother. Deletions were flanked by short direct repeats. We conclude that such patients suffer from a familial form of mitochondrial disease clinically resembling Pearson's syndrome, with a probably autosomal dominant inheritance.

Adult↗
PubMed1994

Vector competence of some neotropical sandflies for the Leishmania (Viannia) braziliensis complex.

To evaluate the vector competence of some Lutzomyia spp. (Diptera: Psychodidae) for Leishmania (Viannia) spp. (Kinetoplastida: Trypanosomatidae), experimental infections of anthropophilic sandflies from the Colombian Pacific coast were performed, through membrane feeding and xenodiagnosis on hamsters infected with Le. (V.)braziliensis or Le. (V.)panamensis. Wild-caught or F1 generation females of Lutzomyia gomezi, Lu. hartmanni, Lu.panamensis and Lu.trapidoi were allowed to feed on hamster lesions and then maintained at 26 degrees C and > 80% r.h. on a sugar-water diet until dissection on the fifth day post-infection (p.i.). Despite similar infection rates (range 37-44%) in both Lu.gomezi and Lu.trapidoi, infections were heavier (> 100 parasites) in the latter species. Infections of Lu.trapidoi with Le.braziliensis (n = 21) and Le.panamensis (n = 27) showed parasite migration toward the foregut, with promastigote colonization of the stomodeal valve and appearance of infective forms. In contrast, infections of Lu.gomezi with Le.braziliensis (n = 10) and Le.panamensis (n = 5) were light (< 50 parasites) and usually restricted to the pylorus. In Lu.hartmanni, only a few promastigotes were found in the pylorus and midgut of 3/8 specimens infected with Le.braziliensis, and no Le.panamensis developed (n = 19). By day 5 p.i., promastigote colonization of the hind- and midgut by Le.panamensis was observed in 2/4 Lu.panamensis but not Le.braziliensis (n = 3). It was concluded that Lu.trapidoi is a more efficient vector than Lu.gomezi for both Le.braziliensis and Le.panamensis, and that Lu.hartmanni and Lu.panamensis are of minor importance for Leishmania transmission in this endemic area.

Animals↗
PubMed1994

Genetic Structure of Rhizobium etli biovar phaseoli Associated with Wild and Cultivated Bean Plants (Phaseolus vulgaris and Phaseolus coccineus) in Morelos, Mexico.

The genetic structure of Rhizobium etli biovar phaseoli was determined for five populations in three different locations in the state of Morelos, Mexico, by using starch gel electrophoresis for five to nine polymorphic loci. Two populations were sampled during two different years from nodules of cultivated and wild common bean plants (Phaseolus vulgaris). The three other populations were associated with wild runner beans (P. coccineus) and sampled during 1988. The Rhizobium populations differ genetically both among sites and among populations within the same site in different years, as shown by differences in allelic frequencies, genetic differentiation analysis, and differences in electrotypes. The total genetic diversity for the five populations during 1988 was H = 0.487; there were also high levels of genetic variation within each population. We found the highest linkage disequilibrium in a global analysis for all the populations. At a local scale, we also found significant linkage disequilibrium in two populations, although the distribution of the D' suggest some recombination at a local scale. The other three rhizobium populations exhibit low linkage disequilibrium. A cluster analysis (UPGMA) of pairwise genetic distances showed that bacteria isolated from most wild Phaseolus spp. are grouped by population, whereas those obtained from cultivated P. vulgaris are very heterogeneous. The analysis of the genetic structure of Rhizobium strains may allow the identification of strains that are naturally well adapted to a wide range of different environments, which may be useful for agricultural purposes or as a starting point for developing improved Rhizobium strains.

Journal Article↗
PubMed1994

Didelphis marsupialis, an important reservoir of Trypanosoma (Schizotrypanum) cruzi and Leishmania (Leishmania) chagasi in Colombia.

The role of Didelphis marsupialis as a reservoir of zoonotic hemoflagellates was examined in two ecologically distinct settings in Colombia. While 72% (12 of 18) of the opossums collected in the tropical rain forest harbored Trypanosoma cruzi, other mammals in the area had lower infection rates: 1.3% (Proechymis semispinosus [spiny rat]; 13% Tylomys mirae [climbing rat]; and 6% Rattus rattus). Trypanosoma cruzi isolates from D. marsupialis were similar to zymodeme 1 (Z1), and two of four phenotypes were shared with Tylomys mirae, which is also predominantly arboreal. Terrestrial (P. semispinosus) and peridomestic (R. rattus) animals were infected with Z3 or other Z1 phenotypes, respectively. Schizodeme analysis showed polymorphisms among isolates from mammals, reflecting diverse modes of transmission, and a complex epidemiologic situation. Despite the lower infection rate of the opossum (14%) found in our study in the tropical dry forest as compared with the tropical wet forest, Chagas' disease has been reported only in the former area. This suggests that the lack of alternative blood sources for triatomines of the tropical dry forest, where mammals are less abundant than in the wet forest, may increase the risk of human infection. Among several species of mammals captured in the tropical dry forest, Leishmania chagasi was isolated from 22.7% (5 of 22) D. marsupialis. This finding confirms the important role of opossums in Colombian foci of visceral leishmaniasis, including those where the phlebotomine species involved in transmission is Lutzomyia evansi, an alternative vector to the more common Lutzomyia longipalpis.

Animals↗
PubMed1993

RNA synthesis in isolated mitochondria from brain cortex, cerebellum and stem: evidence of different transcriptional rates.

1. A system for studying RNA synthesis in isolated sheep brain mitochondria was set up to investigate the transcriptional activity of different brain regions (cortex, cerebellum and brain stem). In this system, mitochondrial DNA is transcribed and RNA processed in a way that faithfully reproduces the in vivo process. 2. The comparison of the electrophoretic patterns of the mitochondrial DNA transcription products showed that although they were qualitatively similar, there were large differences in the rate of mitochondrial DNA transcription of the three regions studied, cerebellum and brain stem showing transcriptional rates which were 34 and 18% respectively of that of cerebral cortex.

Animals↗
PubMed1993

Measurement of HIV virus load and genotypic resistance by gene amplification in asymptomatic subjects treated with combination therapy.

Quantification of viral load in HIV disease has become increasingly important as a marker of antiviral efficacy. We applied gene amplification techniques in vivo to asses antiretroviral activity of combination therapy. Five HIV-infected subjects, four of whom were drug naive, were administered combination therapy with zidovudine (ZDV) and didanosine (ddI). Plasma and peripheral blood mononuclear cells (PBMC) were obtained twice at baseline and then at 1, 3, 6, 9, and 12 months after the initiation of therapy. Results show that plasma HIV RNA copy number fell from 2,170 +/- 660/ml to undetectable at 1 month, with continued suppression at 12 months. HIV proviral DNA copy number decreased from 3.9 to 3.0 log10/10(6) CD4+ T cells at 12 months. Cell dilution cultures were positive in 4 of 5 subjects at baseline and in only 1 of 5 after 12 months. CD4+ T-cell count increased from 390 +/- 30/mm3 pretherapy, to 505 +/- 66/mm3 after 6 months of therapy, but returned to baseline levels after 12 months of therapy. No mutations were detected from PBMC DNA for codon 215 and 74 in the HIV pol gene from the drug-naive subjects. These findings suggest that gene amplification techniques can be used to study changes in viral load or genotype and can be applied in real time to samples from patients involved in clinical trials.

CD4-Positive T-Lymphocytes↗
PubMed1992

Specific increase of a mitochondrial RNA transcript in chronic ethanol-fed rats.

An in vitro transcription system utilizing isolated mitochondria has been used to study the effect of chronic ethanol consumption on liver mitochondrial DNA transcription. The results obtained showed an overall increase of RNA synthesis and a dramatic accumulation of a discrete polyadenylated RNA species. This effect is a consequence of the chronic ethanol consumption since these changes do not occur when isolated control mitochondria are incubated in the presence of ethanol.

Alcoholism↗
PubMed1992

Determination of valproic acid by high-performance liquid chromatography with photodiode-array and fluorescence detection.

The derivatization of valproic acid and undecylenic acid with 4-bromomethyl-7-methoxycoumarin is described. The derivatives were detected by photodiode-array and fluorescence detectors. The optimum monitoring conditions and the stability of the suspension solution and derivatives were investigated. A high-performance liquid chromatographic (HPLC) method with isocratic or gradient elution has been established for the analysis. This method has been used for the determination of total and free valproic acid in serum. There is a satisfactory correlation between the results obtained by this HPLC method and those measured by the enzyme immunoassay. Some other common anti-epileptic drugs did not interfere with the analysis. The method is simple and fast and has better sensitivity and linearity than enzyme immunoassay. It is suited for routine therapeutic drug monitoring.

Chromatography, High Pressure Liquid↗