RESUMEN
Remote Infrared Matrix Assisted Laser Desorption Ionization (Remote IR MALDI) system (SpiderMass) with endogenous water as matrix allows to perform real-time DMPK in vivo. In this work, SpiderMass was used to analyze the impact on metabolite production or release of invalidated pro-protein PC1/3 macrophages by Short RNA (shRNA) versus scramble shRNA with Paclitaxel. Time course in vivo experiments were then performed on the inner and outer faces of patients' forearms or comedo treated with Melascreen (Ducray) containing ascorbyl glucoside. Finally, the impact of car pollution (emitted soot) on skin was also investigated. Taken together, we demonstrate that the SpiderMass instrument opens the door to clinical, pharmaceutical and environmental domains for real-time, in vivo pharmacokinetic (Drug Metabolism and PharmacoKinetics, DMPK) analysis.
Asunto(s)
Macrófagos/metabolismo , Piel/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Animales , Línea Celular , Contaminantes Ambientales/farmacología , Femenino , Humanos , Rayos Láser , Metabolismo de los Lípidos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Masculino , Paclitaxel/farmacología , ARN Interferente Pequeño/genética , Ratas , Piel/efectos de los fármacos , Piel/efectos de la radiación , Envejecimiento de la Piel , Hollín/farmacología , Protectores Solares/farmacología , Urtica dioicaRESUMEN
Endosomal TLR9 is considered as a potent anti-tumoral therapeutic target. Therefore, it is crucial to decipher the mechanisms controlling its trafficking since it determines TLR9 activation and signalling. At present, the scarcity of molecular information regarding the control of this trafficking and signalling is noticeable. We have recently demonstrated that in macrophages, proprotein convertase 1/3 (PC1/3) is a key regulator of TLR4 Myd88-dependent signalling. In the present study, we established that PC1/3 also regulates the endosomal TLR9. Under CpG-ODN challenge, we found that PC1/3 traffics rapidly to co-localize with TLR9 in CpG-ODN-containing endosomes with acidic pH. In PC1/3 knockdown macrophages, compartmentalization of TLR9 was altered and TLR9 clustered in multivesicular bodies (MVB) as demonstrated by co-localization with Rab7. This demonstrates that PC1/3 controls TLR9 trafficking. This clustering of TLR9 in MVB dampened the anti-inflammatory STAT3 signalling pathway while it promoted the pro-inflammatory NF-kB pathway. As a result, macrophages from PC1/3 KO mice and rat PC1/3-KD NR8383 macrophages secreted more pro-inflammatory cytokines such as TNF-α, IL6, IL1α and CXCL2. This is indicative of a M1 pro-inflammatory phenotype. Therefore, PC1/3 KD macrophages represent a relevant mean for cell therapy as "Trojan" macrophages.
Asunto(s)
Proproteína Convertasa 1/metabolismo , Transducción de Señal , Receptor Toll-Like 9/metabolismo , Animales , Citocinas/biosíntesis , Endosomas/metabolismo , Activación Enzimática/efectos de los fármacos , Macrófagos Alveolares/metabolismo , Macrófagos Peritoneales/metabolismo , Ratones , Ratones Noqueados , Ratones Transgénicos , FN-kappa B/metabolismo , Oligodesoxirribonucleótidos/inmunología , Oligodesoxirribonucleótidos/farmacología , Unión Proteica , Transporte de Proteínas , Proteolisis/efectos de los fármacos , Ratas , Factor de Transcripción STAT3/metabolismoRESUMEN
A microcosm study was conducted to evaluate dechlorination of trichloroethene (TCE) to ethene and survival of dechlorinating bacteria after a thermal treatment in order to explore the potential for post-thermal bioremediation. Unamended microcosms containing groundwater and aquifer material from a contaminated site dechlorinated TCE to cis-1,2-dichloroethene (cDCE), while lactate-amended microcosms dechlorinated TCE to cDCE or ethene. A thermal treatment was simulated by heating a sub-set of microcosms to 100 degrees C for 10d followed by cooling to 10 degrees C over 150 d. The heated microcosms demonstrated no dechlorination when unamended. However, when amended with lactate, cDCE was produced in 2 out of 6 microcosms within 300 d after heating. Dechlorination of TCE to cDCE thus occurred in fewer heated (2 out of 12) than unheated (10 out of 12) microcosms. In unheated microcosms, the presence of dechlorinating microorganisms, including Dehalococcoides, was confirmed using nested PCR of 16S rRNA genes. Dechlorinating microorganisms were detected in fewer microcosms after heating, and Dehalococcoides were not detected in any microcosms after heating. Dechlorination may therefore be limited after a thermal treatment in areas that have been heated to 100 degrees C. Thus, inflow of groundwater containing dechlorinating microorganisms and/or bioaugmention may be needed for anaerobic dechlorination to occur after a thermal treatment.
Asunto(s)
Bacterias/metabolismo , Calor , Tricloroetileno/química , Contaminantes Químicos del Agua/aislamiento & purificación , Purificación del Agua/métodos , ADN Bacteriano/análisis , Sedimentos Geológicos/microbiología , Oxidación-Reducción , Reacción en Cadena de la Polimerasa , Tricloroetileno/metabolismoRESUMEN
Interferon alpha (IFNalpha), a type I interferon, can be considered as a viral infection marker because this cytokine is induced during many viral infections. However, it is quite difficult to detect IFNalpha in sera. Investigations are interested in various intra-cellular IFNalpha-induced proteins as viral infection markers. However the activity of these enzymes increased not only in response to type I IFNs but also to type II IFN. MxA protein can be detected in the cytoplasm of IFNalpha/beta-treated cells, whereas other cytokines, including IFNgamma, are poor inducers. Using an immunochemiluminescent assay, we studied MxA protein in whole blood of 34 patients with various viral infections. The whole blood was drawn into sterile vacuum tubes containing heparin or EDTA. MxA values were relatively similar in heparin-treated samples and EDTA-treated samples, with differences not exceeding 1 ng/ml. The levels of MxA protein were compared in whole blood obtained by using two different lysis procedures. A correlation was found between the MxA levels obtained by using procedure I and procedure II, but higher amounts of MxA protein were found with procedure II. The second procedure is rapid and more convenient than the other and it is carried out in one step which reduce technical problems. High levels of MxA protein were found in peripheral blood cells of patients with acute viral infections (Rotavirus, Adenovirus, RSV, CMV), but MxA protein was not elevated in bacterial infections. The MxA levels were also studied in peripheral blood of 32 HCV positive patients. MxA protein was not found in most of IFNalpha-untreated patients, even those with high viral load. In contrast, high levels of MxA protein were found in IFNalpha-treated patients. MxA quantitation can be considered as a specific marker of acute viral infections, and could be useful in the management of treatment with IFNalpha.
Asunto(s)
Proteínas de Unión al GTP , Proteínas/aislamiento & purificación , Virosis/sangre , Hepatitis C/sangre , Humanos , Proteínas de Resistencia a MixovirusRESUMEN
Platelia-Toxo IgA and IMx Toxo IgA assays were used with 260 serum samples, of which 93 were from seroconverted patients, 58 were from 21 congenitally infected children, and 109 were from uninfected patients, to detect anti-P30 immunoglobulin A antibodies. Because of its enhanced sensitivity, Platelia-Toxo IgA is more efficient in diagnosing acute or congenital toxoplasmosis. IMx Toxo IgA must not be used to diagnose congenital toxoplasmosis.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Inmunoensayo/métodos , Inmunoglobulina A/sangre , Toxoplasma/inmunología , Toxoplasmosis/diagnóstico , Animales , Estudios de Evaluación como Asunto , Femenino , Humanos , Inmunoensayo/estadística & datos numéricos , Lactante , Recién Nacido , Embarazo , Complicaciones Parasitarias del Embarazo/diagnóstico , Complicaciones Parasitarias del Embarazo/inmunología , Sensibilidad y Especificidad , Toxoplasmosis/inmunología , Toxoplasmosis Congénita/diagnóstico , Toxoplasmosis Congénita/inmunologíaRESUMEN
It was hypothesized that chronic psychiatric patients who had quit smoking would be more functional and have lower Brief Psychiatric Rating Scale (BPRS) scores than those who continued to smoke. We interviewed 300 chronic psychiatric patients followed in the community. Fourteen percent were former smokers and nearly 11% had never smoked. Fifty-six percent of the sample were current smokers who had no intention of quitting, 13% were considering quitting, and 6% were seriously preparing to quit or had actually quit for a short period. When compared with current smokers, former smokers were more likely to live independently (p < .026) and less likely to have a drug or alcohol problem (p < .013). A random sample of current smokers were compared with former smokers on the BPRS. Former smokers had lower total BPRS scores (p < .03), and lower withdrawal/retardation subscale scores (p < .0058) than current smokers. We concluded that better functioning patients who smoked would be more likely to quit.
Asunto(s)
Trastornos Mentales/diagnóstico , Cese del Hábito de Fumar/psicología , Fumar/epidemiología , Adulto , Anciano , Alcoholismo/epidemiología , Enfermedad Crónica , Comorbilidad , Femenino , Estudios de Seguimiento , Humanos , Masculino , Trastornos Mentales/epidemiología , Trastornos Mentales/psicología , Persona de Mediana Edad , Escalas de Valoración Psiquiátrica , Índice de Severidad de la Enfermedad , Trastornos Relacionados con Sustancias/epidemiologíaRESUMEN
Using a highly discriminatory DNA typing technique, based on the polymerase chain reaction and reverse dot blot hybridization, more refined results were obtained on the association of particular HLA class II alleles, haplotypes and genotypes with insulin-dependent diabetes mellitus in the Belgian population. The previously reported predisposing effect for the DRB1*0301 encoded DR3 serologic specificity was confirmed and could be assigned to the DRB3*0200 encoded DR52b serologic specificity. A second high risk haplotype, DRB1*0401-DQB1*0302 encoding the DR4-DQ8 serologic specificity, accounted for increased susceptibility both in the total insulin-dependent diabetic population and among DR4-positive patients. Moreover, we found that these DR4 associated DRB1 and DQB1 alleles act as independent risk factors. A possible role for the DPB1 locus can be rejected since the observed predisposing effect for DPB1*0202 probably occurred due to linkage disequilibrium of this allele with DRB1*0301. Particular extended haplotypes accounted for the decreased relative risk observed for the DR2, DR11 and DR13 serologic specificities. The highest relative risk was observed for those DQA1/DQB1 genotypes, allowing for the formation of 4SS (DQ alpha Arg52+/DQ beta Asp57-) heterodimers.
Asunto(s)
Diabetes Mellitus Tipo 1/inmunología , Antígenos HLA-D/genética , Adolescente , Adulto , Alelos , Secuencia de Bases , Bélgica , Niño , Preescolar , Cartilla de ADN , Diabetes Mellitus Tipo 1/epidemiología , Diabetes Mellitus Tipo 1/genética , Susceptibilidad a Enfermedades , Femenino , Antígenos HLA-DQ/genética , Antígenos HLA-DR/genética , Haplotipos , Humanos , Lactante , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Valores de Referencia , Factores de RiesgoRESUMEN
A reverse-hybridization assay, the line probe assay (LiPA), based on variations found in the 5' untranslated regions of the different hepatitis C virus (HCV) genotypes was developed, permitting simple and fast determination of four HCV genotypes and their subtypes. Using this assay, 61 PCR-positive Brazilian HCV sera were typed. Of the sera, 33% had a type 1 HCV infection, 38% had type 1b (related to HCV-J), 1.5% had type 2a (related to HC-J6), 24.5% had type 3 (related to E-b1 and HCV-T), and 3% of the sera were co-infected. This assay format was further evaluated using 13 sera from Belgium and the Netherlands, and all of these could be classified. Two pools of Japanese sera were classified as either type 2a or were co-infected with types 1b and 2a, but no type 2b sequences were detected. Another eight PCR-positive sera were obtained from Burundi and Gabon. The sequence of the 5' untranslated region of these African viruses was strongly divergent from the three previously described types. Therefore, these isolates were tentatively classified as type 4. These and some of the other non-type 1 sera often demonstrated weaker reactivities than type 1 isolates in currently used second generation antibody confirmation assays.
Asunto(s)
Sondas de ADN , Hepacivirus/clasificación , Secuencia de Bases , ADN Viral/sangre , Variación Genética , Hepacivirus/genética , Hepacivirus/aislamiento & purificación , Hepatitis C/epidemiología , Humanos , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico/métodos , Reacción en Cadena de la Polimerasa , Sensibilidad y Especificidad , Homología de Secuencia de Ácido Nucleico , Serotipificación/métodos , Terminología como AsuntoRESUMEN
BioArgos (Sanofi Diagnostics Pasteur, Marnes-la-Coquette, France) is a fully automated blood culture system that detects carbon dioxide production by infrared spectroscopy through a glass bottle. This hands-off system was compared with the BACTEC NR-660 system (Becton Dickinson Diagnostic Instrument Systems, Towson, Md.). A total of 336 microorganisms belonging to 74 taxa were tested in simulated blood cultures by both systems. Experimental data showed no significant differences between the two systems. The inclusive detection times (+/- the standard deviations) were 33.2 +/- 28.7 and 35.0 +/- 30.6 h with BioArgos and BACTEC, respectively. Anaerobes were detected earlier with BioArgos, whereas detection of some organisms that need oxygen to grow was slightly delayed. In conclusion, BioArgos is as reliable and accurate as BACTEC NR-660 and shows better practicability owing to noninvasive detection, reduction of vial manipulation, and absence of daily maintenance.
Asunto(s)
Técnicas Bacteriológicas , Sangre/microbiología , Aerobiosis , Anaerobiosis , Técnicas Bacteriológicas/estadística & datos numéricos , Estudios de Evaluación como Asunto , Bacterias Gramnegativas/aislamiento & purificación , Bacterias Grampositivas/aislamiento & purificación , Humanos , Micología/métodos , Sensibilidad y Especificidad , Levaduras/aislamiento & purificaciónRESUMEN
Three oligonucleotide probe sequences were inferred from the 16S ribosomal ribonucleic acid (rRNA) and the 16S-23S rRNA spacer sequences of Bordetella pertussis ATCC 10380. These probes were used in hybridization tests with deoxyribonucleic acid from Bordetella species and other relevant bacterial taxa. A probe from the spacer region hybridized exclusively to the B. pertussis strains tested and not to strains from other species. Using a combination of three probes, B. pertussis, B. parapertussis/B. bronchiseptica and B. avium could be specifically identified and differentiated from other taxa. Differentiation between B. parapertussis and B. bronchiseptica was not possible with the probes used. Using the spacer probe, a colorimetric hybridization assay specific for B. pertussis was developed based on enzymatic amplification of the 16S-23S rRNA spacer and reverse hybridization in microtitre wells. As compared with results using agarose gel electrophoresis, and Southern and dot-spot hybridization with a 32P-labelled probe, this assay proved to be faster and easier to perform and was found to be at least as sensitive and specific.
Asunto(s)
Bordetella pertussis/genética , Bordetella/genética , ADN Bacteriano/genética , Hibridación de Ácido Nucleico , Secuencia de Bases , Sondas de ADN , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genética , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: To determine the in vitro antimicrobial susceptibility of gonococcal strains isolated in 1988 among female prostitutes in Kinshasa, Zaire and to characterise strains with high level tetracycline resistance. METHODS: Minimal inhibitory concentrations of 8 antimicrobials were measured by agar dilution technique. Plasmid-profiles and serovars were determined. RESULTS: Two hundred and thirteen strains of Neisseria gonorrhoeae were tested of which 59% were beta-lactamase producers and an additional 21% showed intermediate or chromosomal resistance to penicillin (MIC = 0.5-8 mg/l). Eleven percent of the strains were resistant to the combination sulfamethoxazole-trimethoprim (MIC greater than 8 mg/l) and 57% of the isolates showed decreased susceptibility to thiamphenicol (MIC = 1-4 mg/l). All strains were sensitive to spectinomycin, norfloxacin and ceftriaxone and moderately sensitive to kanamycin. Chromosomal resistance to tetracycline was observed in 45% of strains (MIC = 2-8 mg/l). Ten percent were highly resistant to tetracycline (TRNG, MIC = 16-128 mg/l) and were shown to carry a plasmid borne Tet M determinant; such strains were not found in Kinshasa in 1985. TRNG belonged to 4 different serovars, which were also the dominant serovars in non-TRNG. CONCLUSION: These findings illustrate the high frequency of multiresistant gonococci in Zaire and suggest that high level tetracycline resistant strains of N. gonorrhoeae have become endemic in Central Africa.
Asunto(s)
Antibacterianos/farmacología , Neisseria gonorrhoeae/efectos de los fármacos , Plásmidos/genética , Resistencia a la Tetraciclina/genética , Southern Blotting , República Democrática del Congo , Electroforesis en Gel de Agar , Femenino , Humanos , Técnicas In Vitro , Pruebas de Sensibilidad Microbiana , Neisseria gonorrhoeae/genética , Trabajo SexualRESUMEN
With the aim of achieving earlier diagnosis of congenital toxoplasmosis, anti-P30 immunoglobulin A (IgA) antibodies were assayed by using a Platelia-Toxo IgA kit with samples from 72 children born to mothers who seroconverted during pregnancy. A total of 148 serum samples and 1 cerebrospinal fluid samples were from 23 congenitally infected children (2 serum samples were collected from fetuses), and 74 serum samples were from 49 uninfected children. Among the 23 infected children, anti-P30 IgA antibodies were present in all infants either at birth or in the following weeks, whereas anti-P30 IgM antibodies were present in 13 from the 23 infected children either at birth or in the following weeks. Serum samples collected in utero from two infected children were also tested. One of these samples was positive for both anti-P30 IgA and anti-P30 IgM antibodies, whereas both children were negative at birth for these antibodies. Neither anti-P30 IgA nor anti-P30 IgM antibodies were detected in 47 of 49 uninfected children. These results suggest that detection of anti-P30 IgA antibodies by the Platelia-Toxo IgA kit is a very effective method for early diagnosis of congenital toxoplasma infection.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Inmunoensayo/métodos , Inmunoglobulina A/análisis , Toxoplasmosis Congénita/diagnóstico , Animales , Estudios de Evaluación como Asunto , Femenino , Sangre Fetal/inmunología , Humanos , Inmunoglobulina M/análisis , Recién Nacido , Masculino , Embarazo , Diagnóstico Prenatal , Toxoplasma/inmunología , Toxoplasmosis Congénita/inmunologíaRESUMEN
Part of a ribosomal ribonucleic acid (rRNA) cistron of Haemophilus ducreyi was enzymically amplified using conserved primers within the rRNA molecules, cloned in a plasmid vector, and sequenced. From the nucleotide sequence, eight oligonucleotides complementary to different regions in the 16S and 23S rRNA molecules were selected, chemically synthesized, and used as hybridization probes. Hybridization experiments with at least 41 H. ducreyi strains and 13 or 14 non-H. ducreyi strains revealed that all eight oligonucleotide probes were highly reliable and completely specific for H. ducreyi strains. Comparisons of 16S rRNA sequences confirm that H. ducreyi is a member of the Pasteurellaceae though not closely related to other species in this family.
Asunto(s)
Haemophilus ducreyi/clasificación , Sondas de Oligonucleótidos , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genética , Secuencia de Bases , Chancroide/microbiología , Clonación Molecular , Haemophilus ducreyi/genética , Humanos , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , ARN Bacteriano/genética , Alineación de SecuenciaRESUMEN
The reliability of an rRNA-derived oligonucleotide probe for Neisseria gonorrhoeae was tested with 187 N. gonorrhoeae isolates, 81 Neisseria meningitidis isolates, and several strains of other bacterial species. The probe proved to be 100% specific and 100% sensitive. N. gonorrhoeae cells could also be reliably identified in contaminated cultures with the oligonucleotide probe. The 2.6-megadalton cryptic plasmid used as a probe for N. gonorrhoeae was shown to be less sensitive, detecting 179 of 181 N. gonorrhoeae isolates.
Asunto(s)
Sondas de ADN , Neisseria gonorrhoeae/genética , ARN Bacteriano/genética , Estudios de Evaluación como Asunto , Gonorrea/diagnóstico , Humanos , Técnicas de Sonda Molecular , Neisseria gonorrhoeae/aislamiento & purificación , Neisseria meningitidis/genética , Neisseria meningitidis/aislamiento & purificación , Plásmidos , ARN Ribosómico/genética , Especificidad de la EspecieRESUMEN
We report a new case of Chediak-Higashi disease successfully treated by the transplantation of allogeneic bone marrow. Recurrent infections led to the diagnosis of the disease at the age of 15 months. At two and a half years of age, during a phase of accelerated disease activity, the patient received a bone marrow transplant donated by an HLA-identical brother. The patient was conditioned by chemotherapy alone; T-cells were removed from the graft and cyclosporin A was given to prevent graft-versus-host disease. Evidence of acceptance of the transplant was apparent 14 days after the procedure. Two months after the transplant, the blood count was normal, NK activity was satisfactory and no evidence of GVH disease was present. Incomplete hematopoietic chimerism was found (with two erythrocyte and lymphocyte populations). After four years follow-up, the patient is doing well and has no infections or evidence of active disease.
Asunto(s)
Trasplante de Médula Ósea , Síndrome de Chediak-Higashi/terapia , Preescolar , Humanos , MasculinoRESUMEN
Providing preventive services to patients is a priority for most family physicians. In this study, a three-item preventive checklist reminded patients 65 years of age and older to obtain an annual blood pressure check, influenza immunization, and cancer checkup. Over a one-year period, patients who received the checklist (n = 59) obtained 46% of indicated cancer detection services, while those who did not (n = 55) obtained 30%. Both groups had similar rates of obtaining blood pressure measurement (over 90%) and influenza immunization (over 45%). Patients themselves may be an underutilized resource for improving preventive care.