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1.
Interv Neuroradiol ; 28(4): 381-385, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34515565

RESUMO

Transradial access is widely used in cardiological adult interventions and less in pediatrics. In recent years, this access has become more popular in the neuroradiological community in adult patients since it has fewer complications and is more comfortable for the patient after the procedure. We present a single-center case series of 52 transradial access neurointerventions (43 angiographies and 9 therapeutic procedures) in pediatric patients, with a failure of 4 cases (7.7%) in which we could not puncture the artery, crossing over to transfemoral access. Since in five cases we did angiography followed by therapeutic intervention, thus doing only one puncture access for both procedures, then our access failure rate was 10.6%. The 34 successful transradial access solely angiographies had a median radiation exposure of 887 mGy (interquartile range 628-1352), median fluoroscopy time of 9.5 min (interquartile range 7.5-15.3), and median procedure time of 28 min (interquartile range 24-33 min) Therapeutic procedure diagnosis were: one ruptured saccular aneurysm, two juvenile nasopharyngeal angiofibromas, and five arteriovenous malformations. The transradial access neurointerventions for pediatric population older than 11 years is safe and feasible, having previous experience in adults. Younger population should be considered on a case-to-case basis, depending on ultrasound measurement of the arterial diameter and the materials available.


Assuntos
Procedimentos Neurocirúrgicos , Artéria Radial , Adolescente , Angiografia/efeitos adversos , Angiografia/métodos , Estudos de Viabilidade , Humanos , Procedimentos Neurocirúrgicos/efeitos adversos , Procedimentos Neurocirúrgicos/métodos , Artéria Radial/diagnóstico por imagem , Artéria Radial/cirurgia , Resultado do Tratamento
2.
ISME J ; 15(1): 78-92, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32879462

RESUMO

Helicobacter pylori is a common component of the human stomach microbiota, possibly dating back to the speciation of Homo sapiens. A history of pathogen evolution in allopatry has led to the development of genetically distinct H. pylori subpopulations, associated with different human populations, and more recent admixture among H. pylori subpopulations can provide information about human migrations. However, little is known about the degree to which some H. pylori genes are conserved in the face of admixture, potentially indicating host adaptation, or how virulence genes spread among different populations. We analyzed H. pylori genomes from 14 countries in the Americas, strains from the Iberian Peninsula, and public genomes from Europe, Africa, and Asia, to investigate how admixture varies across different regions and gene families. Whole-genome analyses of 723 H. pylori strains from around the world showed evidence of frequent admixture in the American strains with a complex mosaic of contributions from H. pylori populations originating in the Americas as well as other continents. Despite the complex admixture, distinctive genomic fingerprints were identified for each region, revealing novel American H. pylori subpopulations. A pan-genome Fst analysis showed that variation in virulence genes had the strongest fixation in America, compared with non-American populations, and that much of the variation constituted non-synonymous substitutions in functional domains. Network analyses suggest that these virulence genes have followed unique evolutionary paths in the American populations, spreading into different genetic backgrounds, potentially contributing to the high risk of gastric cancer in the region.


Assuntos
Infecções por Helicobacter , Helicobacter pylori , América , Europa (Continente) , Variação Genética , Genoma Bacteriano , Helicobacter pylori/genética , Humanos , Estados Unidos , Virulência/genética
3.
Molecules ; 25(23)2020 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-33260521

RESUMO

Cryptocarya alba (Peumo; CA) and Laurelia sempervirens (Laurel; LS) are herbs native to the Chilean highlands and have historically been used for medicinal purposes by the Huilliches people. In this work, the essential oils were extracted using hydrodistillation in Clevenger apparatus and analyzed by GC-MS to determine their composition. The antioxidant capacity (AC) was evaluated in vitro. The cytotoxicity was determined using cell line cultures both non tumoral and tumoral. The toxicity was determined using the nematode Caenorhabditis elegans. The antimicrobial activity was evaluated against 52 bacteria using the agar disc diffusion method and the minimum inhibitory concentrations (MICs) were determined. The principal compounds found in C. alba essential oil (CA_EO) were α-terpineol (24.96%) and eucalyptol (21.63%) and were isazafrol (91.9%) in L. sempervirens essential oil (LS_EO). Both EOs showed antioxidant capacity in vitro. Both EO showed antibacterial activity against bacteria using. LS_EO showed more inhibitory effect on these cell lines respect to CA_EO. Both EOs showed toxicity against the nematode C.elegans at 3.12-50 mg/mL. The essential oils of CA and LS have an important bioactive potential in their antioxidant, antibacterial and cytotoxicity activity. Both essential oils could possibly be used in the field of natural medicine, natural food preservation, cosmetics, sanitation and plaguicides among others.


Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Cryptocarya/química , Óleos Voláteis/farmacologia , Extratos Vegetais/farmacologia , Animais , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/crescimento & desenvolvimento , Proliferação de Células , Humanos , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Células Tumorais Cultivadas
4.
Sex Transm Dis ; 47(5S Suppl 1): S61-S65, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32004258

RESUMO

BACKGROUND: In a recent sexually transmitted disease surveillance report from the Centers for Disease Control and Prevention, Miami-Dade County had the nation's fourth highest rate of infectious syphilis, with rates of congenital syphilis on the rise. As a public health response, Homestead Hospital, in collaboration with the Florida Department of Health, enhanced their emergency department's routine HIV/HCV "opt-out" screening infrastructure to include a syphilis smart screening algorithm. The purpose of this article is to describe the development, implementation, and evaluation of the algorithm. METHODS: A retrospective evaluation of patient records prompted the development of the algorithm. Homestead Hospital's electronic health record system automatically triggers a syphilis test based on the reason for medical visit (e.g., rash, penile discharge, a positive pregnancy test, historical or present sexually transmitted disease result). If a patient tests positive, he/she is counseled and linked to care. RESULTS: Since implementation (April 2018 to August 2019), the smart screening algorithm triggered 4806 syphilis tests: 122 patients tested positive (2.5% seropositivity). After confirmatory testing, 59 patients were positive for syphilis, of which 27 were pregnant. CONCLUSIONS: The Homestead Hospital and Department of Health-Miami-Dade's response to Miami-Dade County's syphilis problem is innovative and replicable. The program embraces technology, enhances the routine opt-out screening model, and does not affect preexisting workflows. Ultimately, implementation of this algorithm allows patients to get treatment, receive comprehensive prevention services, and, in some cases, avert congenital syphilis.


Assuntos
Epidemias/prevenção & controle , Programas de Rastreamento/estatística & dados numéricos , Sorodiagnóstico da Sífilis/métodos , Sífilis/diagnóstico , Algoritmos , Feminino , Florida/epidemiologia , Humanos , Masculino , Programas de Rastreamento/métodos , Estudos Retrospectivos , Sífilis/epidemiologia , Sífilis/prevenção & controle
5.
Hum Vaccin Immunother ; 11(3): 776-88, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25750999

RESUMO

The Neisseria meningitidis outer membrane protein PorA from a Chilean strain was purified as a recombinant protein. PorA mixed with AbISCO induced bactericidal antibodies against N. meningitidis in mice. When PorA was fused to the Helicobacter pylori HpaA antigen gene, the specific response against H. pylori protein increased. Splenocytes from PorA-immunized mice were stimulated with PorA, and an increase in the secretion of IL-4 was observed compared with that of IFN-γ. Moreover, in an immunoglobulin sub-typing analysis, a substantially higher IgG1 level was found compared with IgG2a levels, suggesting a Th2-type immune response. This study revealed a peculiar behavior of the purified recombinant PorA protein per se in the absence of AbISCO as an adjuvant. Therefore, the resistance of PorA to proteolytic enzymes, such as those in the gastrointestinal tract, was analyzed, because this is an important feature for an oral protein adjuvant. Finally, we found that PorA fused to the H. pylori HpaA antigen, when expressed in Lactococcus lactis and administered orally, could enhance the antibody response against the HpaA antigen approximately 3 fold. These observations strongly suggest that PorA behaves as an effective oral adjuvant.


Assuntos
Adesinas Bacterianas/imunologia , Adjuvantes Imunológicos/administração & dosagem , Anticorpos Antibacterianos/sangue , Helicobacter pylori/imunologia , Porinas/imunologia , Adesinas Bacterianas/administração & dosagem , Adesinas Bacterianas/genética , Adjuvantes Imunológicos/genética , Administração Oral , Animais , Feminino , Imunoglobulina G/sangue , Interferon gama/metabolismo , Interleucina-4/metabolismo , Lactococcus lactis/genética , Lactococcus lactis/metabolismo , Leucócitos Mononucleares/imunologia , Camundongos Endogâmicos BALB C , Porinas/administração & dosagem , Porinas/genética , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Soro/química , Baço/imunologia
6.
Mucosal Immunol ; 6(5): 950-959, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23299619

RESUMO

Helicobacter pylori induces less gastric inflammation in children than adults. Here we investigated whether this reduced inflammation involves dysregulated T helper type 17 (Th17) responses. H. pylori-infected children and adults in Santiago, Chile had similar levels of H. pylori colonization, proportions of bacteria containing cagA and s1/s2 vacA markers of virulence, and strain genotypes (predominantly hpEurope), but the children had significantly reduced levels of gastric inflammation and neutrophil infiltration. The reduced neutrophil accumulation in the infected children was accompanied by significantly fewer gastric Th17 cells and significantly lower levels of interleukin (IL)-17-specific mRNA and protein compared with the infected adults. The gastric mucosa of H. pylori-infected children also contained higher numbers of IL-10+ cells and increased levels of both IL-10 and Foxp3 mRNA compared with that of the infected adults. Thus, reduced gastric inflammation, including diminished neutrophil accumulation, in H. pylori-infected children compared with infected adults is likely due to downregulated gastric Th17/IL-17 responses as a consequence of enhanced mucosal regulatory T-cell activity in the children.


Assuntos
Mucosa Gástrica/imunologia , Gastrite/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Neutrófilos/imunologia , Linfócitos T Reguladores/imunologia , Células Th17/imunologia , Adolescente , Adulto , Criança , Progressão da Doença , Feminino , Fatores de Transcrição Forkhead/metabolismo , Gastrite/etiologia , Regulação da Expressão Gênica , Infecções por Helicobacter/complicações , Helicobacter pylori/patogenicidade , Humanos , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-17/genética , Interleucina-17/metabolismo , Masculino , Pessoa de Meia-Idade , Fatores de Virulência
7.
Cell Tissue Res ; 342(3): 423-35, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21042919

RESUMO

Hemocyanins are copper-containing glycoproteins in some molluscs and arthropods, and their best-known function is O(2) transport. We studied the site of their biosynthesis in the gastropod Concholepas concholepas by using immunological and molecular genetic approaches. We performed immunohistochemical staining of various organs, including the mantle, branchia, and hepatopancreas, and detected C. concholepas hemocyanin (CCH) molecules in circulating and tissue-associated hemocytes by electron microscopy. To characterize the hemocytes, we purified them from hemolymph. We identified three types of granular cells. The most abundant type was a phagocyte-like cell with small cytoplasmic granules. The second type contained large electron-dense granules. The third type had vacuoles containing hemocyanin molecules suggesting that synthesis or catabolism occurred inside these cells. Our failure to detect cch-mRNA in hemocytes by reverse transcription with the polymerase chain reaction (RT-PCR) led us to propose that hemocytes instead played a role in CCH metabolism. This hypothesis was supported by colloidal gold staining showing hemocyanin molecules in electron-dense granules inside hemocytes. RT-PCR analysis, complemented by in situ hybridization analyses with single-stranded antisense RNAs as specific probes, demonstrated the presence of cch-mRNA in the hepatopancreas; this was consistent with the specific hybridization signal and confirmed the hepatopancreas as the site of CCH synthesis. Finally, we investigated the possibility that CCH catabolism in hemocytes was involved in the host immune response and in the generation of secondary metabolites such as antimicrobial peptides and phenoloxidase.


Assuntos
Gastrópodes/metabolismo , Hemocianinas/biossíntese , Hemócitos/metabolismo , Hepatopâncreas/metabolismo , Animais , Imunofluorescência , Hibridização In Situ , Microscopia Eletrônica , RNA Antissenso , Reação em Cadeia da Polimerase Via Transcriptase Reversa
8.
World J Gastroenterol ; 16(3): 320-9, 2010 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-20082477

RESUMO

AIM: To evaluate how widely Helicobacter pylori (H. pylori) HopE and HopV porins are expressed among Chilean isolates and how seroprevalent they are among infected patients in Chile. METHODS: H. pylori hopE and hopV genes derived from strain CHCTX-1 were cloned by polymerase chain reaction (PCR), sequenced and expressed in Escherichia coli AD494 (DE3). Gel-purified porins were used to prepare polyclonal antibodies. The presence of both genes was tested by PCR in a collection of H. pylori clinical isolates and their expression was detected in lysates by immunoblotting. Immune responses against HopE, HopV and other H. pylori antigens in sera from infected and non-infected patients were tested by Western blotting using these sera as first antibody on recombinant H. pylori antigens. RESULTS: PCR and Western blotting assays revealed that 60 and 82 out of 130 Chilean isolates carried hopE and hopV genes, respectively, but only 16 and 9, respectively, expressed these porins. IgG serum immunoreactivity evaluation of 69 H. pylori-infected patients revealed that HopE and HopV were infrequently recognized (8.7% and 10.1% respectively) compared to H. pylori VacA (68.1%) and CagA (59.5%) antigens. Similar values were detected for IgA serum immunoreactivity against HopE (11.6%) and HopV (10.5%) although lower values for VacA (42%) and CagA (17.4%) were obtained when compared to the IgG response. CONCLUSION: A scarce expression of HopE and HopV among Chilean isolates was found, in agreement with the infrequent seroconversion against these antigens when tested in infected Chilean patients.


Assuntos
Infecções por Helicobacter/metabolismo , Helicobacter pylori/isolamento & purificação , Helicobacter pylori/metabolismo , Porinas/metabolismo , Anticorpos Antibacterianos/sangue , Chile , Genótipo , Humanos , Fenótipo , Porinas/genética , Estudos Retrospectivos
9.
J Cell Biochem ; 107(6): 1083-96, 2009 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-19479899

RESUMO

The mechanisms of peroxisomal biogenesis remain incompletely understood, specially regarding the role of the endoplasmic reticulum (ER) in human cells, where genetic disorders of peroxisome biogenesis lead to Zellweger syndrome (ZS). The Pex3p peroxisomal membrane protein (PMP) required for early steps of peroxisome biogenesis has been detected in the ER in yeast but not in mammalian cells. Here, we show that Pex3p-GFP expressed in a new ZS cell line (MR), which lacks peroxisomes due to a mutation in the PEX3 gene, localizes first in the ER and subsequently in newly formed peroxisomes. Pex3p bearing an artificial N-glycosylation site shows an electrophoretic shift indicative of ER targeting while en route to preformed peroxisomes in normal fibroblast. A signal peptide that forces its entry into the ER does not eliminate its capability to drive peroxisome biogenesis in ZS cells. Thus, Pex3p is able to drive peroxisome biogenesis from the ER and its ER pathway is not privative of ZS cells. Cross-expression experiments of Pex3p in GM623 cells lacking Pex16p or Pex16p in MR cells lacking Pex3p, showed evidence that Pex3p requires Pex16p for ER location but is dispensable for the ER location of Pex16p. These results indicate that Pex3p follows the ER-to-peroxisomal route in mammalian cells and provides new clues to understand its function.


Assuntos
Retículo Endoplasmático/metabolismo , Lipoproteínas/fisiologia , Proteínas de Membrana/fisiologia , Peroxissomos/metabolismo , Aciltransferases , Estudos de Casos e Controles , Catalase , Retículo Endoplasmático/enzimologia , Fibroblastos/citologia , Humanos , Lipoproteínas/genética , Lipoproteínas/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mutação , Peroxinas , Transporte Proteico , Síndrome de Zellweger
10.
Biol Res ; 40(2): 231-49, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-18064360

RESUMO

Peroxisomes are thought to be formed by division of pre-existing peroxisomes after the import of newly synthesized proteins. However, it has been recently suggested that the endoplasmic reticulum (ER) provides an alternative de novo mechanism for peroxisome biogenesis in some cells. To test a possible role of the ER-Golgi transit in peroxisome biogenesis in mammalian cells, we evaluated the biogenesis of three peroxisomal membrane proteins (PMPs): ALDRP (adrenoleukodystrophy related protein), PMP70 and Pex3p in CHO cells. We constructed chimeric genes encoding these PMPs and green fluorescent protein (GFP), and transiently transfected them to wild type and mutant CHO cells, in which normal peroxisomes were replaced by peroxisomal membrane ghosts. The expressed proteins were targeted to peroxisomes and peroxisomal ghosts correctly in the presence or absence of Brefeldin A (BFA), a drug known to block the ER-Golgi transit. Furthermore, low temperature did not disturb the targeting of Pex3p-GFP to peroxisomes. We also constructed two chimeric proteins of PMPs containing an ER retention signal "DEKKMP": GFP-ALDRP-DEKKMP and myc- Pex3p-DEKKMP. These proteins were mostly targeted to peroxisomes. No colocalization with an ER maker was found. These results suggest that the classical ER-Golgi pathway does not play a major role in the biogenesis of mammalian PMPs.


Assuntos
Retículo Endoplasmático/fisiologia , Complexo de Golgi/fisiologia , Proteínas de Membrana/metabolismo , Mutação , Peroxissomos/metabolismo , Animais , Células CHO , Cricetinae , Cricetulus , Retículo Endoplasmático/metabolismo , Proteínas de Membrana/genética
11.
Cancer Epidemiol Biomarkers Prev ; 16(4): 662-7, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17416755

RESUMO

BACKGROUND AND AIMS: Chile ranks fifth in the world among countries with the highest incidence of gastric cancer. The aim was to quantify the association between Helicobacter pylori infection and gastric cancer mortality at the county of residence. METHODS: A cross-sectional household survey, a probability sample of the Chilean adult population, provided 2,615 participants in whom serum H. pylori IgG antibodies were measured (ELISA). The spatial pattern of 48,367 deaths due to gastric cancer which occurred from 1985 to 2002 was analyzed using a hierarchical Poisson regression model; 333 counties were categorized as low, medium, and high gastric cancer mortality with median gastric cancer death rates of 11.4, 19.1, and 26.0 per 100,000 inhabitants, respectively. The association between H. pylori positivity and gastric cancer mortality in the county of residence was assessed by multivariate Poisson regression for complex samples. RESULTS: H. pylori prevalence was 73.0% [95% confidence intervals (CI), 70.0-76.0], higher in men [prevalence rate ratio (PRR), 1.1 (95% CI, 1.01-1.20)], peaked at ages 45 to 64, and dropped after age 65. It was higher among residents in counties with high gastric cancer mortality (79.7%; 95% CI, 76.4-82.6) compared to counties with low gastric cancer mortality (62.3%; 95% CI, 53.8-70.2; corresponding PRR, 1.3; 95% CI, 1.1-1.5); under age 24, H. pylori infection was 79.7% (95% CI, 72.2-85.6) versus 39.8% (95% CI, 19.6-64.2) among residents in counties with high and low gastric cancer mortalities, respectively (PRR, 2.0; 95% CI, 1.1-3.7). CONCLUSIONS: The high prevalence of H. pylori at younger ages was associated with high gastric cancer mortality in the base population.


Assuntos
Infecções por Helicobacter/epidemiologia , Helicobacter pylori , Neoplasias Gástricas/microbiologia , Neoplasias Gástricas/mortalidade , Adolescente , Adulto , Idoso , Distribuição de Qui-Quadrado , Chile/epidemiologia , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Feminino , Helicobacter pylori/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Distribuição de Poisson , Prevalência , Sensibilidade e Especificidade
12.
Microbes Infect ; 9(4): 428-34, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17336120

RESUMO

Helicobacter pylori infection is highly prevalent in Chile (73%). Usually a minority of infected patients develops complications such as ulcers and gastric cancer that have been associated with the presence of virulence factors (cagA, vacA) and host T helper response (Th1/Th2). Our aim was to evaluate the relationship between strain virulence and host immune response, using a multiple regression approach for the development of a model based on data collected from H. pylori infected patients in Chile. We analyzed levels of selected cytokines determined by ELISA (interleukin (IL)-12, IL-10, interferon (IFN)-gamma and IL-4) and the presence of cagA and vacA alleles polymorphisms determined by PCR in antral biopsies of 41 patients referred to endoscopy. By multiple regression analysis we established a correlation between bacterial and host factors using clinical outcome (gastritis and duodenal ulcer) as dependent variables. The selected model was described by: clinical outcome=0.867491 (cagA)+0.0131847 (IL-12/IL-10)+0.0103503 (IFN-gamma/IL-4) and it was able to explain over 90% of clinical outcomes observations (R(2)=96.4). This model considers that clinical outcomes are better explained by the interaction of host immune factors and strain virulence as a complex and interdependent mechanism.


Assuntos
Citocinas/imunologia , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/virologia , Helicobacter pylori/imunologia , Helicobacter pylori/patogenicidade , Fatores de Virulência/genética , Adolescente , Adulto , Alelos , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Criança , Pré-Escolar , Úlcera Duodenal/imunologia , Úlcera Duodenal/virologia , Feminino , Gastrite/imunologia , Gastrite/virologia , Helicobacter pylori/genética , Humanos , Masculino , Pessoa de Meia-Idade , Células Th1/imunologia , Células Th2/imunologia
13.
Biol. Res ; 40(2): 231-249, 2007. ilus
Artigo em Inglês | LILACS | ID: lil-468194

RESUMO

Peroxisomes are thought to be formed by division of pre-existing peroxisomes after the import of newly synthesized proteins. However, it has been recently suggested that the endoplasmic reticulum (ER) provides an alternative de novo mechanism for peroxisome biogenesis in some cells. To test a possible role of the ER-Golgi transit in peroxisome biogenesis in mammalian cells, we evaluated the biogenesis of three peroxisomal membrane proteins (PMPs): ALDRP (adrenoleukodystrophy related protein), PMP70 and Pex3p in CHO cells. We constructed chimeric genes encoding these PMPs and green fluorescent protein (GFP), and transiently transfected them to wild type and mutant CHO cells, in which normal peroxisomes were replaced by peroxisomal membrane ghosts. The expressed proteins were targeted to peroxisomes and peroxisomal ghosts correctly in the presence or absence of Brefeldin A (BFA), a drug known to block the ER-Golgi transit. Furthermore, low temperature did not disturb the targeting of Pex3p-GFP to peroxisomes. We also constructed two chimeric proteins of PMPs containing an ER retention signal "DEKKMP": GFP-ALDRP-DEKKMP and myc- Pex3p-DEKKMP. These proteins were mostly targeted to peroxisomes. No colocalization with an ER maker was found. These results suggest that the classical ER-Golgi pathway does not play a major role in the biogenesis of mammalian PMPs.


Assuntos
Animais , Cricetinae , Retículo Endoplasmático/fisiologia , Complexo de Golgi/fisiologia , Mutação , Proteínas de Membrana/metabolismo , Peroxissomos/metabolismo , Células CHO , Cricetulus , Retículo Endoplasmático/metabolismo , Proteínas de Membrana/genética
14.
J Clin Endocrinol Metab ; 89(2): 946-51, 2004 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-14764819

RESUMO

Steroidogenic acute regulatory protein (StAR) plays a crucial role in the transport of cholesterol from the cytoplasm to the inner mitochondrial membrane, facilitating its conversion to pregnenolone by cytochrome P450scc. Its essential role in steroidogenesis was demonstrated after observing that StAR gene mutations gave rise to a potentially lethal disease named congenital lipoid adrenal hyperplasia, in which virtually no steroids are produced. We report here a 2-month-old female patient, karyotype 46XY, who presented with growth failure, convulsions, dehydration, hypoglycemia, hyponatremia, hypotension, and severe hyperpigmentation suggestive of adrenal insufficiency. Serum cortisol, 17OH-progesterone, dehydroepiandrosterone sulfate, testosterone, 17OH-pregnenolone, and aldosterone levels were undetectable in the presence of high ACTH and plasma renin activity levels. Immunohistochemical analysis of testis tissues revealed the absence of StAR protein. Molecular analysis of StAR gene demonstrated a homozygous G to T mutation within the splice donor site of exon 1 (IVS1 + 1G>T). Her parents and one brother were heterozygous for this mutation. In vitro analysis of the mutation was performed in COS cells transfected with minigenes coding regions spanning exon-intron 1 to 3 carrying the mutant and the wild-type sequences. RT-PCR analyses of the mutant gene showed an abnormal mRNA transcript of 2430 bp (normal size 433 bp). Sequence analysis of the mutant mRNA demonstrated the retention of intron 1. Immunolocalization of the StAR minigene product detected the peptide in the mitochondria of COS cells transfected with the wild-type minigene but not in those transfected with the mutant minigene. We conclude that this mutation gives rise to a truncated StAR protein, which lacks an important N-terminal region and the entire lipid transfer domain.


Assuntos
Hiperplasia Suprarrenal Congênita/genética , Mutação , Fosfoproteínas/genética , Splicing de RNA , Hiperplasia Suprarrenal Congênita/metabolismo , Hiperplasia Suprarrenal Congênita/patologia , Animais , Sequência de Bases/genética , Células COS , Chlorocebus aethiops , Enzimas de Restrição do DNA , Feminino , Expressão Gênica , Humanos , Imuno-Histoquímica , Lactente , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Distribuição Tecidual
15.
Biol Res ; 35(3-4): 333-8, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12462985

RESUMO

Intra-oviductal administration of RNA obtained from oviducts of estradiol-treated rats resulted in accelerated egg transport (Ríos et al., 1997). It is probable that estradiol-induced messenger RNA (mRNA) entered oviductal cells and was translated into the proteins involved in accelerated egg transport. In order to test this interpretation we deposited in vivo 50 micrograms of pure beta-galactosidase (beta-gal) mRNA, 50 micrograms of pure DNA from the reporter gene beta-gal under SV40 promoter or the vehicle (control oviducts) into the oviductal lumen of rats. Twenty four hours later the beta-gal activity was assayed in oviductal tissue homogenates using o-nitrophenyl-beta-D-galactopyranoside as a substrate. The administration of beta-gal mRNA and pSVBgal plasmid increased beta-gal activity by 71% and 142%, respectively, over the control oviducts. These results indicate that naked DNA and mRNA coding for beta-gal can enter oviductal cells and be translated into an active enzyme. They are consistent with the interpretation that embryo transport acceleration caused by the injection of estradiol-induced RNA in the oviduct involves translation of the injected mRNA.


Assuntos
Tubas Uterinas/enzimologia , RNA Mensageiro/metabolismo , Transcrição Gênica , Transfecção/métodos , beta-Galactosidase/metabolismo , Animais , Feminino , Transporte do Óvulo , Plasmídeos/administração & dosagem , RNA Mensageiro/administração & dosagem , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Ribonucleases/análise , Estatísticas não Paramétricas , beta-Galactosidase/genética
16.
Biol Res ; 35(1): 67-84, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12125208

RESUMO

We have cloned and sequenced ten Helicobacter pylori genes from a Chilean strain (CH-CTX1) including: a cytotoxin VacA fragment, a CagA fragment (A17), a species-specific protein (TsaA), urease subunits (UreA, UreB), a flagellin subunit (FlaB), heat shock proteins (HspA and HspB), adhesin (HpaA) and a lipoprotein (Lpp20). We compared their deduced amino acid sequences with the corresponding sequences from three unrelated H. pylori strains, including fully sequenced strains 26695(UK) and J99(USA), and found that eight of them (UreA, UreB, FlaB, HspA, HspB, Lpp20, TsaA and HpaA) presented more than 97.3% identity. In contrast, VacA partial sequence showed lower identity values (93.2-94.9%). Moreover, we found major differences in the A17 region respect to the number and arrangement of the internal repeated elements when sequences from different strains were aligned. The A17 regions from strains CH-CTX1 and 26695 are very similar (91.8% identity) but lacked 6 repeated elements when compared to the Australian strains ATCC 43526 and NCTC 11637. The CCUG 17874 A17 region showed the largest deletion involving 9 repeats. A17 size differences between strains CCUG 17874 and CH-CTX1 were verified by PCR and polypeptide size. Such differences may explain variations in virulence among H. pylori strains as well as diversity in serum immunoreactivity.


Assuntos
Antígenos de Bactérias , Proteínas de Bactérias/genética , Variação Genética , Helicobacter pylori/genética , Alelos , Sequência de Aminoácidos , Sequência de Bases , Western Blotting/métodos , Proteínas de Transporte/genética , Clonagem Molecular , Primers do DNA , Genes , Helicobacter pylori/patogenicidade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Virulência/genética
17.
Biol. Res ; 35(3/4): 333-338, 2002. ilus, graf
Artigo em Inglês | LILACS | ID: lil-339726

RESUMO

Intra-oviductal administration of RNA obtained from oviducts of estradiol-treated rats resulted in accelerated egg transport (Rios et al., 1997). It is probable that estradiol-induced messenger RNA (mRNA) entered oviductal cells and was translated into the proteins involved in accelerated egg transport. In order to test this interpretation we deposited in vivo 50 micrograms of pure beta-galactosidase (beta-gal) mRNA, 50 micrograms of pure DNA from the reporter gene beta-gal under SV40 promoter or the vehicle (control oviducts) into the oviductal lumen of rats. Twenty four hours later the beta-gal activity was assayed in oviductal tissue homogenates using o-nitrophenyl-beta-D-galactopyranoside as a substrate. The administration of beta-gal mRNA and pSVBgal plasmid increased beta-gal activity by 71% and 142%, respectively, over the control oviducts. These results indicate that naked DNA and mRNA coding for beta-gal can enter oviductal cells and be translated into an active enzyme. They are consistent with the interpretation that embryo transport acceleration caused by the injection of estradiol-induced RNA in the oviduct involves translation of the injected mRNA


Assuntos
Animais , Feminino , Ratos , beta-Galactosidase , Tubas Uterinas , Transporte do Óvulo , RNA Mensageiro , beta-Galactosidase , Tubas Uterinas , Expressão Gênica , Ratos Sprague-Dawley
18.
Biol. Res ; 35(1): 67-84, 2002. ilus, tab
Artigo em Inglês | LILACS | ID: lil-309739

RESUMO

We have cloned and sequenced ten Helicobacter pylori genes from a Chilean strain (CH-CTX1) including: a cytotoxin VacA fragment, a CagA fragment (A17), a species-specific protein (TsaA), urease subunits (UreA, UreB), a flagellin subunit (FlaB), heat shock proteins (HspA and HspB), adhesin (HpaA) and a lipoprotein (Lpp20). We compared their deduced amino acid sequences with the corresponding sequences from three unrelated H. pylori strains, including fully sequenced strains 26695(UK) and J99(USA), and found that eight of them (UreA, UreB, FlaB, HspA, HspB, Lpp20, TsaA and HpaA) presented more than 97.3% identity. In contrast, VacA partial sequence showed lower identity values (93.2-94.9%). Moreover, we found major differences in the A17 region respect to the number and arrangement of the internal repeated elements when sequences from different strains were aligned. The A17 regions from strains CH-CTX1 and 26695 are very similar (91.8% identity) but lacked 6 repeated elements when compared to the Australian strains ATCC 43526 and NCTC 11637. The CCUG 17874 A17 region showed the largest deletion involving 9 repeats. A17 size differences between strains CCUG 17874 and CH-CTX1 were verified by PCR and polypeptide size. Such differences may explain variations in virulence among H. pylori strains as well as diversity in serum immunoreactivity.


Assuntos
Proteínas de Bactérias , Clonagem Molecular , Variação Genética , Helicobacter pylori , Alelos , Sequência de Aminoácidos , Sequência de Bases , Western Blotting , Primers do DNA , Genes , Helicobacter pylori , Reação em Cadeia da Polimerase , Virulência
19.
Rev. Inst. Nac. Enfermedades Respir ; 6(4): 223-7, oct.-dic. 1993. tab, ilus
Artigo em Espanhol | LILACS | ID: lil-134885

RESUMO

La proteinosis alveolar (PA), es una enfermedad pulmonar difusa caracterizada por el depósito de material fosfolipídico en los espacios alveolares, con conservación de la arquitectura pulmonar y sin reacción inflamatoria. La etiología es desconocida, sin embargo, actualmente, se han demostrado alteraciones anatómicas y funcionales de los macrófagos alveolares. Las evolución de la enfermedad es crónica y en muchas ocasiones indistinguible de enfermedades intersticiales difusas, en algunos casos el diagnóstico es siguerido por su asociación con enfermedades infecciosas (micosis). El diagnóstico definitivo siempre es histopatológico. No existe un tratamiento curativo, sin embargo, en la catualidad se ha utilizado el lavado pulmonar total, como una alternativa con buenos resultados en la mayoría de los casos. El presente reporte es con motivo de 2 casos estudiados en el Instituto con diferencias en su presentación clínica, radiológica, funcional y evolución final


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Proteinose Alveolar Pulmonar/etiologia , Líquido da Lavagem Broncoalveolar/terapia , Diagnóstico Diferencial , Macrófagos/imunologia , Proteinose Alveolar Pulmonar/diagnóstico , Proteinose Alveolar Pulmonar/fisiopatologia
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