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N Delgado

Publications and source records attributed to N Delgado.

14 recordsLinked to original sources

Profiling gene expression in Coccidioides posadasii.

Coccidioides posadasii is a dimorphic fungal pathogen which grows as a filamentous saprobe in the soil and multicellular parasitic form in host lung tissue. Studies of gene expression profiles during saprobic and parasitic phase development can provide clues about morphogenetic regulation and may lead to the discovery of molecular targets for novel antifungal drugs. Suppression-subtractive hybridization (SSH) and quantitative real-time polymerase chain reaction (QRT-PCR) were used to identify and quantify differential gene expression during in vitro growth of Coccidioides. DNA fragments obtained from the subtraction of cDNA pools derived from the saprobic and parasitic phase RNA preparations were each cloned into an appropriate vector and subjected to sequence analysis. Semi-quantitative, reverse transcription polymerase chain reaction (RT-PCR) experiments were first conducted to assess whether these inserts represented differentially expressed genes. Nucleotide sequences of the partial and full-length genes selected by RT-PCR were obtained by genome walking and rapid amplification of cDNA ends (RACE) methods. QRT-PCR analysis of the expression of these genes during saprobic and parasitic cell growth was then conducted using DNA standard curves normalized to a constitutively expressed control gene. Four C. posadasii genes whose expression is essentially restricted to the parasitic cycle were discovered using this approach. These genes include homologues of OPS1 (encodes opsin-related protein), MDR1 (multidrug resistance protein), ALDR1 (aldehyde reductase), and PSP1 (hypothetical lipid transporter/flippase protein). The combined applications of SSH and QRT-PCR permit global analysis of gene expression patterns in C. posadasii.

Coccidioides↗

Presence of a pertussis toxin-sensitive G protein alpha subunit in Sporothrix schenckii.

As an initial step in the study of the role of G proteins in signal transduction in Sporothrix schenckii, we identified a Galphai subunit using different experimental approaches. Western blots of fungal membrane preparations using anti-Galphacommon and anti-Galphai1-Galphai2 antibodies identified a band of approximately 41 kDa. Pertussis toxin-catalyzed adenosine diphosphate (ADP)-ribosylation of these membrane fractions confirmed the presence of a protein substrate of 41 kDa. A 357 bp polymerase chain reaction (PCR) product obtained using fungal DNA as template and primers targeted to conserved Galphai sequences, was used as a probe to isolate a clone from an S. schenckii genomic library. A partial sequence for a Galphai subunit was obtained from this clone. The sequence was completed using the rapid amplification of cDNA ends (RACE) technique with mycelium and yeast cDNA. The cDNA sequence revealed a 1059 bp open reading frame encoding a 353 amino acid Galphai subunit of 41 kDa, more than 90% identical to the CPG-1 of Cryphonectria parasitica, and GNA-1 of Neurospora crassa. The genomic sequence was obtained by PCR using fungal DNA, and revealed a 1250 bp sequence and the presence of three introns. These results provide evidence for the first time of the presence and expression of a Galphai homolog in a pathogenic dimorphic fungus.

Amino Acid Sequence↗

[Stratification of a hyperendemic city in hemorrhagic dengue].

Any effort to control dengue hemorrhagic fever (DHF) faces a number of challenges. Among these are the great environmental heterogeneity of homes and neighborhoods in urban centers where the primary dengue vector, Aedes aegypti, breeds, as well as shortages of resources and of personnel trained in mosquito control. Adequate epidemiological surveillance could serve as a basis to begin to stratify urban communities and identify the areas in them where control efforts should be focused. In this study we stratified Maracay, Venezuela, a city with hyperendemic dengue hemorrhagic fever, using a geographic information system and analyzing the persistence, incidence, and prevalence of dengue, by means of clinical diagnoses reported from 1993 through 1998. Maracay has around one million inhabitants living in some 349 neighborhoods in the six communities that make up the greater Maracay metropolitan area. During that 1993-1998 period the Maracay area reported 10,576 cases of dengue, 2,593 cases of DHF, and 8 deaths. The incidence of DHF was related to the incidence of dengue, the number of inhabitants in an area, and population density. The spatial pattern of dengue incidence was stable over the years that were studied, and significant, positive relationships were found between pairs of years and the incidence of dengue by neighborhood. The persistence of dengue was related directly to monthly incidence by neighborhood. These spatial patterns helped to divide the city into three strata: 68 neighborhoods without apparent dengue, 226 neighborhoods with low persistence and prevalence, and 55 neighborhoods with high persistence and prevalence. We recommend giving the highest priority for control efforts to these 55 neighborhoods, which make up just 35% of the Maracay urban area but had 70% of all the reported dengue cases.

Aedes↗

Clinical effectiveness of essential oil-containing dentifrices in controlling oral malodor.

PURPOSE: To assess the efficacy of two essential oil-containing dentifrices in the reduction of human intrinsic oral malodor ("morning bad breath"). MATERIALS AND METHODS: Two clinical trials were conducted which used observer-blind, negative control, parallel designs. To insure a homogeneous test population with similar oral odor characteristics, subjects followed instructions that included toothbrushing with a standard ADA-accepted fluoride dentifrice the night before. On odor evaluation days, subjects refrained from all oral hygiene procedures and from eating or drinking any food or beverage or smoking prior to and during the odor evaluations. Eighty healthy adult subjects in the first trial and 90 in the second trial with qualifying baseline levels of oral malodor participated. Odor ratings of each subject's breath were made by 4-5 trained judges using a nine point hedonic scale, i.e., 1 = most pleasant, 5 = neutral, 9 = most unpleasant. Qualifying subjects were randomly assigned one of two essential oil dentifrices or a negative control dentifrice. Subjects brushed with their assigned dentifrice for 60 s, rinsed with 20 ml of water for 10 s, and received post-treatment hedonic odor ratings at 30, 60, 90, 120, 180, and 240 min. Data were analyzed using an ANCOVA to adjust for baseline scores. RESULTS: The essential oil dentifrices were significantly more effective (P < or = 0.033) than the control in reducing intrinsic oral malodor from 90 to 120 min.

Adult↗

Is fibrin glue useful in preventing dialysate leakage in children on CAPD? Preliminary results of a prospective randomized study.

Children on continuous ambulatory peritoneal dialysis (CAPD) have a higher incidence of dialysate leakage than patients receiving cycler dialysis. To date, fibrin glue has been used to treat dialysate leakage but not as a method of prevention. Therefore, we conducted a prospective study in which 20 catheters were implanted in 19 children, with each catheter randomly assigned to either the treatment group (1 mL of fibrin glue added to peritoneal cuff suture) or the control group. There was no difference in mean patient age, elapsed time between catheter implantation and first catheter usage, and albumin concentration between the two groups. Dialysate leakage was detected in 4 patients in the control group, 3 of whom underwent early initiation of dialysis, but none of the patients in the treatment group (including 4 with early dialysis) had a similar problem. In summary, these preliminary data suggest that fibrin glue may be of benefit when applied at the time of catheter implantation. Further experience is necessary before this method can be routinely recommended.

Adolescent↗

[Several new concepts on the terminal mechanism of mammal fertilization].

We briefly reviewed some ionic mechanisms participating in the fulfillment of the acrosome reaction processes, indicating the results obtained by the simultaneous and dynamic quantitation of sodium, potassium, calcium and hydrogen concentrations in the in incubation medium of human spermatozoa induced to achieve the acrosome reaction through the addition of cAMP and follicular fluid. At the same time, the participation of zinc and some enzymatic activities, like phospholipase A2, in the occurrence of this process, is indicated. A study is done about the sperm nuclear decondensation mechanisms, pointing at the importance of various participants in this event, like the disulfide groups reducing agents, some metalic ions and the glycosaminoglycanes, as well as a proposal of a mechanism which could be physiologically functional for the in vivo occurrence of this phenomenon. Finally we present some results explaining the DNA synthesis activation, which is repressed in the spermatozoa since the spermiogenesis final stages, and is indispensable for the chromosomic duplication required during egg segmentation.

Acrosome↗

[New concepts about the terminal mechanism of fertilization in mammals].

We briefly reviewed some ionic mechanisms participating in the fulfillment of the acrosome reaction processes, indicating the results obtained by the simultaneous and dynamic quantitation of sodium, potassium, calcium and hydrogen concentrations in the in incubation medium of human spermatozoa induced to achieve the acrosome reaction through the addition of cAMP and follicular fluid. At the same time, the participation of zinc and some enzymatic activities, like phospholipase A2, in the occurrence of this process, is indicated. A study is done about the sperm nuclear decondensation mechanisms, pointing at the importance of various participants in this event, like the disulfide groups reducing agents, some metal ions and the glycosaminoglycans, as well as a proposal of a mechanism which could be physiologically functional for the in vivo occurrence of this phenomenon. Finally we present some results explaining the DNA synthesis activation, which is repressed in the spermatozoa since the spermiogenesis final stages, and is indispensable for the chromosomic duplication required during egg segmentation.

Acrosome↗

Effect of cupric ions on the initiation protein synthesis rate in the human endometrium.

The effect of cupric ions on the initiation protein synthesis rate of the human endometrium was studied. Addition of copper to the complete ribosomal system decreased the binding of [3H]Met-tRNA(i) to the isolated ribosomes with a plateau at about 70% inhibition with concentrations higher than 150 microM. The initiation activity was GTP-dependent with a maximum at 2 mM. This activity was very rapid, requiring 5 min to complete the reaction. Incubation of isolated initiation factors with copper (300 microM) inhibited the formation of the ternary complex. When the complete system was reconstituted with salt-washed ribosomes after ternary complex formation, no significant change on the inhibition pattern was observed. Addition of initiation factors to 5-min preincubated salt-washed ribosomes with 300 microM copper, after the elimination of excess copper, induced only a 12% decrease on Met-tRNA(i) binding. This effect was not modified by the presence of Sparsomycin, an elongation inhibitor. It was concluded that copper interferes with the initiation process, probably at the ternary complex formation level.

Adult↗

[Association of Wilms' tumor, pseudohermaphroditism and glomerular nephropathy. Partial and complete forms of the syndrome].

Four patients, followed at the Buenos Aires Cildren's Hospital are reported: one with a nephrotic syndrome and male pseudohermaphroditism, the second with nephrotic syndrome, male pseudohermaphroditism and Wilms' tumor, the third with pseudohermaphroditism and Wilms' tumor and the fourth with a nephrotic syndrome and Wilms' tumor. The cases presenting nephrotic syndrome, all had an early onset and a rapid and fatal course leading to death in renal failure or secondary to a related bacterial infection. The renal histology in all of the cases with nephrotic syndrome, was very similar: diffuse involvement of all glomeruli consisting in a severe increase of the mesangial matrix, with scarce mesengial proliferation.

Adult↗