RESUMEN
AIMS: To determine whether oral antioxidant therapies, of various types and doses, are able to prevent or treat women with preeclampsia. DATA SYNTHESIS: The following databases were searched: MEDLINE, CENTRAL, LILACS, and Web of Science. Inclusion criteria were: a) randomized clinical trials; b) oral antioxidant supplementation; c) study in pregnant women; d) control group, treated or not with placebo. Papers were excluded if they evaluated antioxidant nutrient supplementation associated with other non-antioxidant therapies. Data were extracted and the risk of bias of each study was assessed. Heterogeneity was analyzed using the Cochran Q test, and I2 statistics and pre-specified sensitivity analyses were performed. Meta-analyses were conducted on prevention and treatment studies, separately. The primary outcome was the incidence of preeclampsia in prevention trials, and of perinatal death in treatment trials. Twenty-nine studies were included in the analysis, 19 for prevention and 10 for treatment. The antioxidants used in these studies were vitamins C and E, selenium, l-arginine, allicin, lycopene and coenzyme Q10, none of which showed beneficial effects on the prevention of preeclampsia (RR: 0.89, CI 95%: [0.79-1.02], P = 0.09; I2 = 39%, P = 0.04) and other outcomes. The antioxidants used in the treatment studies were vitamins C and E, N-acetylcysteine, l-arginine, and resveratrol. A beneficial effect was found in intrauterine growth restriction. CONCLUSIONS: Antioxidant therapy had no effects in the prevention of preeclampsia but did show beneficial effects in intrauterine growth restriction, when used in the treatment of this condition.
Asunto(s)
Antihipertensivos/administración & dosificación , Antioxidantes/administración & dosificación , Presión Sanguínea/efectos de los fármacos , Preeclampsia/tratamiento farmacológico , Preeclampsia/prevención & control , Administración Oral , Adolescente , Adulto , Antihipertensivos/efectos adversos , Antioxidantes/efectos adversos , Medicina Basada en la Evidencia , Femenino , Retardo del Crecimiento Fetal/prevención & control , Humanos , Incidencia , Preeclampsia/mortalidad , Preeclampsia/fisiopatología , Embarazo , Ensayos Clínicos Controlados Aleatorios como Asunto , Factores de Riesgo , Resultado del Tratamiento , Adulto JovenRESUMEN
Several works have shown that triterpenes induce peripheral antinociception by activation of cannabinoid receptors and endocannabinoids; besides, several research groups have reported activation of cannabinoid receptors in peripheral antinociception. The aim of this study was to assess the involvement of the cannabinoid system in the antinociceptive effect induced by tingenone against hyperalgesia evoked by prostaglandin E2 (PGE2) at peripheral level. The paw pressure test was used and the hyperalgesia was induced by intraplantar injection of PGE2 (2 µg/paw). All drugs were injected subcutaneously in the hind paws of male Swiss mice. Tingenone (200 µg/paw) administered into the right hind paw induced a local antinociceptive effect, that was antagonized by AM630, a selective antagonist to CB2 cannabinoid receptor. AM251, a selective antagonist to CB1 cannabinoid receptor, did not alter the peripheral antinociceptive effect of tingenone. MAFP, a fatty acid amide hydrolase (FAAH) inhibitor; VDM11, an anandamide reuptake inhibitor; and JZL184, monoacylglycerol lipase (MAGL) inhibitor did not potentiate the peripheral antinociceptive effect of the lower dose of tingenone (50 µg/paw). The results suggest that tingenone induced a peripheral antinociceptive effect via cannabinoid receptor activation. Therefore, this study suggests a pharmacological potential for a new analgesic drug.
Asunto(s)
Analgésicos/farmacología , Agonistas de Receptores de Cannabinoides/farmacología , Triterpenos Pentacíclicos/farmacología , Receptor Cannabinoide CB1/metabolismo , Receptor Cannabinoide CB2/metabolismo , Triterpenos/farmacología , Amidohidrolasas , Animales , Ácidos Araquidónicos/metabolismo , Ácidos Araquidónicos/farmacología , Benzodioxoles/farmacología , Cannabinoides/metabolismo , Endocannabinoides/metabolismo , Hiperalgesia/tratamiento farmacológico , Hiperalgesia/metabolismo , Indoles/farmacología , Masculino , Ratones , Monoacilglicerol Lipasas/metabolismo , Piperidinas/farmacología , Alcamidas Poliinsaturadas/metabolismo , Pirazoles/farmacologíaRESUMEN
The appearance of circulating islet-specific autoantibodies before disease diagnosis is a hallmark of human type 1 diabetes (T1D), and suggests a role for B cells in the pathogenesis of the disease. Alterations in the peripheral B cell compartment have been reported in T1D patients; however, to date, such studies have produced conflicting results and have been limited by sample size. In this study, we have performed a detailed characterization of the B cell compartment in T1D patients (n = 45) and healthy controls (n = 46), and assessed the secretion of the anti-inflammatory cytokine interleukin (IL)-10 in purified B cells from the same donors. Overall, we found no evidence for a profound alteration of the B cell compartment or in the production of IL-10 in peripheral blood of T1D patients. We also investigated age-related changes in peripheral B cell subsets and confirmed the sharp decrease with age of transitional CD19(+) CD27(-) CD24(hi) CD38(hi) B cells, a subset that has recently been ascribed a putative regulatory function. Genetic analysis of the B cell compartment revealed evidence for association of the IL2-IL21â T1D locus with IL-10 production by both memory B cells (P = 6·4 × 10(-4) ) and islet-specific CD4(+) T cells (P = 2·9 × 10(-3) ). In contrast to previous reports, we found no evidence for an alteration of the B cell compartment in healthy individuals homozygous for the non-synonymous PTPN22â Trp(620) T1D risk allele (rs2476601; Arg(620) Trp). The IL2-IL21 association we have identified, if confirmed, suggests a novel role for B cells in T1D pathogenesis through the production of IL-10, and reinforces the importance of IL-10 production by autoreactive CD4(+) T cells.
Asunto(s)
Subgrupos de Linfocitos B/inmunología , Subgrupos de Linfocitos B/metabolismo , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/inmunología , Adolescente , Adulto , Factores de Edad , Autoanticuerpos/inmunología , Estudios de Casos y Controles , Niño , Citocinas/biosíntesis , Diabetes Mellitus Tipo 1/metabolismo , Femenino , Citometría de Flujo , Regulación de la Expresión Génica , Estudios de Asociación Genética , Humanos , Inmunofenotipificación , Masculino , Fenotipo , Proteína Tirosina Fosfatasa no Receptora Tipo 22/genética , Transducción de Señal , Adulto JovenRESUMEN
This study aimed to evaluate the effect of human saliva in vitro and salivary flow in situ on the roughness and mineral content of bleached enamel. Dental specimens were divided into five groups (n=15): not bleached (NB); bleached (35% hydrogen peroxide) and exposed to distilled water (DW); human saliva in vitro (IV); normal salivary flow in situ (NSF); and low salivary flow (LSF) in situ. Enamel roughness (Ra, Rz) and calcium/phosphorus contents were evaluated with laser profilometry and energy-dispersive spectroscopy, respectively, at baseline (T1), after bleaching (T2), and after seven days (T3). Salivary pH and buffer capacity were evaluated with colorimetric strips and salivary calcium and phosphorus with absorbance spectrophotometry. Data were analyzed with non-parametric tests and linear regression (α=0.05). After contact with saliva, Ra and Rz of LSF=DW>IV=NSF=NB was found. For DW and LSF, the roughness of T1Asunto(s)
Blanqueadores Dentales
, Blanqueamiento de Dientes
, Humanos
, Blanqueamiento de Dientes/métodos
, Blanqueadores Dentales/química
, Calcio/análisis
, Peróxido de Hidrógeno/química
, Esmalte Dental
, Minerales/análisis
, Minerales/farmacología
, Fósforo
RESUMEN
Aedes aegypti control is achieved with chemical insecticides that can promote insecticide resistance. In the search for new forms of control, the use of botanical products is currently growing and many tests with oils have already been performed. The plant diversity of Araripe National Forest enables the study of several species against this vector. To evaluate the larvicidal effect of essential oils from plants of this forest, we used field rosemary, copaiba, bay leaf, cashew and pequi. The work was divided into three stages: all oils with the same dosage; the best oil at dosages of 0, 5, 10, 20, 50 and 75 µg/mL; and the best dosage at temperatures of 15, 20, 25, 30 and 35 °C. The oils of field rosemary, copaiba, bay leaf, cashew and pequi were good insecticides when used at dosages above 5 µg/mL. The bay leaf oil showed high larvicidal activity at all dosages tested, showing the highest efficiency at 75 µg/mL. Temperatures of 15 and 35 °C increased the susceptibility of the insect to the effect of the bay leaf oil. The essential oils of field rosemary, copaiba, bay leaf, cashew and pequi, from Araripe National Forest, applied at a dosage of 5µg/mL, showed insecticidal action, although with low efficiency.
Asunto(s)
Aedes , Insecticidas , Aceites Volátiles , Animales , Aceites Volátiles/farmacología , Insecticidas/farmacología , Larva , Mosquitos Vectores , Aceites de Plantas/farmacologíaRESUMEN
Onychomycosis is the most common disease affecting the nail unit and accounts for at least 50% of all nail diseases. In addition, Candida albicans is responsible for approximately 70% of onychomycoses caused by yeasts. This study investigated the antifungal effect of (R) and (S)-citronellal enantiomers, as well as its predictive mechanism of action on C. albicans from voriconazole-resistant onychomycoses. For this purpose, in vitro broth microdilution and molecular docking techniques were applied in a predictive and complementary manner to the mechanisms of action. The main results of this study indicate that C. albicans was resistant to voriconazole and sensitive to the enantiomers (R) and (S)-citronellal at a dose of 256 and 32 µg/mL respectively. In addition, there was an increase in the minimum inhibitory concentration (MIC) of the enantiomers in the presence of sorbitol and ergosterol, indicating that these molecules possibly affect the integrity of the cell wall and cell membrane of C. albicans. Molecular docking with key biosynthesis proteins and maintenance of the fungal cell wall and plasma membrane demonstrated the possibility of (R) and (S)-citronellal interacting with two important enzymes: 1,3-ß-glucan synthase and lanosterol 14α-demethylase. Therefore, the findings of this study indicate that the (R) and (S)-citronellal enantiomers are fungicidal on C. albicans from onychomycoses and probably these substances cause damage to the cell wall and cell membrane of these micro-organisms possibly by interacting with enzymes in the biosynthesis of these fungal structures.
Asunto(s)
Antifúngicos , Onicomicosis , Voriconazol , Candida albicans , Simulación del Acoplamiento MolecularRESUMEN
In this study, a pharmacological approach, together with the paw pressure test, was used to investigate the role of dopamine and its receptors in the peripheral processing of the nociceptive response in mice. Initially, the administration of dopamine (5, 20, and 80 ng/paw) in the hind paw of male Swiss mice (30-40 g) promoted antinociceptive effects in a dose-dependent manner. This was considered a peripheral effect, as it did not produce changes in the nociceptive threshold of the contralateral paw. The D2, D3, and D4 dopamine receptor antagonists remoxipride (4 µg/paw), U99194 (16 µg/paw), and L-745,870 (16 µg/paw), respectively, reversed the dopamine-mediated antinociception in mice with PGE2-induced hyperalgesia. The D1 and D5 dopamine receptor antagonists SKF 83566 (2 µg/paw) and SCH 23390 (1.6 µg/paw), respectively, did not alter dopamine antinociception. In contrast, dopamine at higher doses (0.1, 1, and 10 µg/paw) caused hyperalgesia in the animals, and the D1 and D5 receptor antagonists reversed this pronociceptive effect (10 µg/paw), whereas the D2 receptor antagonist remoxipride did not. Our data suggest that dopamine has a dual effect that depends on the dose, as it causes peripheral antinociceptive effects at small doses via the activation of D2-like receptors and nociceptive effects at higher doses via the activation of D1-like receptors.
Asunto(s)
Analgesia , Dopamina , Analgésicos/efectos adversos , Animales , Antagonistas de Dopamina/farmacología , Hiperalgesia/tratamiento farmacológico , Masculino , Ratones , Nocicepción , Dolor/inducido químicamente , Dolor/tratamiento farmacológico , Receptores de Dopamina D1 , Remoxiprida/efectos adversosRESUMEN
Interest in antiviral plant species has grown exponentially and some have been reported to have anti-HIV properties. This research aims to perform the bio-guided phytochemical fractionation by antiretroviral activity of Lafoensia pacari stem barks. This in vitro experimental study involved the preparation of plant material, obtention of ethanolic extract, fractionation, purification, identification and quantification of fractions, acid-base extraction, nuclear magnetic resonance, HIV-1 RT inhibition test and molecular docking studies. From the bio-guided fractionation by the antiretroviral activity there was a higher activity in the acetanolic subfractions, highlighting the acetate subfraction - neutrals with 60.98% of RT inhibition and ellagic acid with 88.61% of RT inhibition and absence of cytotoxicity. The macrophage lineage cytotoxicity assay showed that the chloroform fraction was more toxic than the acetate fraction. The analysis of the J-resolved spectrum in the aromatic region showed a singlet at 7.48 and 6.93 ppm which was identified as ellagic acid and gallic acid, respectively. The 5TIQ enzyme obtained better affinity parameter with the ellagic acid ligand, which was confirmed by the HSQC-1H-13C spectra. Gallic acid was also favorable to form interaction with the 5TIQ enzyme, being confirmed through the HSQC-1H-13C spectrum. From the PreADMET evaluation it was found that ellagic acid is a promising molecule for its RT inhibition activity and pharmacokinetic and toxicity parameters.
Asunto(s)
Infecciones por VIH , Lythraceae , Acetatos , Ácido Elágico/farmacología , Ácido Gálico/farmacología , Lythraceae/química , Simulación del Acoplamiento Molecular , Extractos Vegetales/toxicidadRESUMEN
Kahweol is a compound derived from coffee with reported antinociceptive effects. Based on the few reports that exist in the literature regarding the mechanisms involved in kahweol-induced peripheral antinociceptive action, this study proposed to investigate the contribution of the endocannabinoid system to the peripheral antinociception induced in rats by kahweol. Hyperalgesia was induced by intraplantar injection of prostaglandin E2(PGE2) and was measured with the paw pressure test. Kahweol and the drugs to test the cannabinoid system were administered locally into the right hind paw. The endocannabinoids were purified by open-bed chromatography on silica and measured by LC-MS. Kahweol (80 µg/paw) induced peripheral antinociception against PGE2-induced hyperalgesia. This effect was reversed by the intraplantar injection of the CB1 cannabinoid receptor antagonist AM251 (20, 40, and 80 µg/paw), but not by the CB2 cannabinoid receptor antagonist AM630 (100 µg/paw). Treatment with the endocannabinoid reuptake inhibitor VDM11 (2.5 µg/paw) intensified the peripheral antinociceptive effect induced by low-dose kahweol (40 µg/paw). The monoacylglycerol lipase (MAGL) inhibitor, JZL184 (4 µg/paw), and the dual MAGL/fatty acid amide hydrolase (FAAH) inhibitor, MAFP (0.5 µg/paw), potentiated the peripheral antinociceptive effect of low-dose kahweol. Furthermore, kahweol increased the levels of the endocannabinoid anandamide, but not of the other endocannabinoid 2-arachidonoylglycerol nor of anandamide-related N-acylethanolamines, in the plantar surface of the rat paw. Our results suggested that kahweol induced peripheral antinociception via anandamide release and activation of CB1 cannabinoid receptors and this compound could be used to develop new drugs for pain relief.
Asunto(s)
Diterpenos , Endocannabinoides , Analgésicos/farmacología , Animales , Café , Hiperalgesia/inducido químicamente , Hiperalgesia/tratamiento farmacológico , Ratas , Receptor Cannabinoide CB1 , Receptor Cannabinoide CB2RESUMEN
Trypanosoma (Megatrypanum) theileri from cattle and trypanosomes of other artiodactyls form a clade of closely related species in analyses using ribosomal sequences. Analysis of polymorphic sequences of a larger number of trypanosomes from broader geographical origins is required to evaluate the clustering of isolates as suggested by previous studies. Here, we determined the sequences of the spliced leader (SL) genes of 21 isolates from cattle and 2 from water buffalo from distant regions of Brazil. Analysis of SL gene repeats revealed that the 5S rRNA gene is inserted within the intergenic region. Phylogeographical patterns inferred using SL sequences showed at least 5 major genotypes of T. theileri distributed in 2 strongly divergent lineages. Lineage TthI comprises genotypes IA and IB from buffalo and cattle, respectively, from the Southeast and Central regions, whereas genotype IC is restricted to cattle from the Southern region. Lineage TthII includes cattle genotypes IIA, which is restricted to the North and Northeast, and IIB, found in the Centre, West, North and Northeast. PCR-RFLP of SL genes revealed valuable markers for genotyping T. theileri. The results of this study emphasize the genetic complexity and corroborate the geographical structuring of T. theileri genotypes found in cattle.
Asunto(s)
Enfermedades de los Bovinos/epidemiología , Bovinos/parasitología , Filogenia , ARN Lider Empalmado/genética , Trypanosoma/clasificación , Trypanosoma/genética , Tripanosomiasis , Animales , Secuencia de Bases , Brasil/epidemiología , Búfalos/parasitología , Enfermedades de los Bovinos/parasitología , ADN Protozoario/análisis , ADN Protozoario/genética , ADN Ribosómico/análisis , ADN Ribosómico/genética , ADN Espaciador Ribosómico/análisis , ADN Espaciador Ribosómico/genética , Evolución Molecular , Genotipo , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , ARN Ribosómico/genética , ARN Ribosómico 5S/genética , Análisis de Secuencia de ADN , Trypanosoma/aislamiento & purificación , Tripanosomiasis/epidemiología , Tripanosomiasis/parasitología , Tripanosomiasis/veterinariaRESUMEN
Science teaching in most Brazilian Universities tends to focus mainly on lectures and provides few opportunities for the development of modern teaching skills. Our group developed an online tool called Adopt a Bacterium, which consists on a Facebook group where teacher assistants (TAs) can interact with students and have a first contact with student-focused learning approaches. This work shows the TAs' own assessment of how the tool could be further explored to help them develop skills and become better teachers.
Asunto(s)
Educación a Distancia/métodos , Ciencia/educación , Formación del Profesorado/métodos , Enseñanza , Brasil , Curriculum , Humanos , UniversidadesRESUMEN
In this study, using a combined data set of SSU rDNA and gGAPDH gene sequences, we provide phylogenetic evidence that supports clustering of crocodilian trypanosomes from the Brazilian Caiman yacare (Alligatoridae) and Trypanosoma grayi, a species that circulates between African crocodiles (Crocodilydae) and tsetse flies. In a survey of trypanosomes in Caiman yacare from the Brazilian Pantanal, the prevalence of trypanosome infection was 35% as determined by microhaematocrit and haemoculture, and 9 cultures were obtained. The morphology of trypomastigotes from caiman blood and tissue imprints was compared with those described for other crocodilian trypanosomes. Differences in morphology and growth behaviour of caiman trypanosomes were corroborated by molecular polymorphism that revealed 2 genotypes. Eight isolates were ascribed to genotype Cay01 and 1 to genotype Cay02. Phylogenetic inferences based on concatenated SSU rDNA and gGAPDH sequences showed that caiman isolates are closely related to T. grayi, constituting a well-supported monophyletic assemblage (clade T. grayi). Divergence time estimates based on clade composition, and biogeographical and geological events were used to discuss the relationships between the evolutionary histories of crocodilian trypanosomes and their hosts.
Asunto(s)
Caimanes y Cocodrilos/parasitología , Evolución Biológica , ADN Ribosómico/genética , Gliceraldehído-3-Fosfato Deshidrogenasa (Fosforilante)/genética , Filogenia , Trypanosomatina/clasificación , África , Animales , América del Sur , Trypanosomatina/citología , Trypanosomatina/aislamiento & purificaciónRESUMEN
The class Cephalopoda (Phylum Mollusca), encompassing squids and octopuses, contains multiple species that are characterized by the presence of specialized organs known to emit light. These complex organs have a variety of morphological characteristics ranging from groups of simple, light-producing cells, to highly specialized organs (light organs) with cells surrounded by reflectors, lenses, light guides, color filters, and muscles. Bacteriogenic light organs have been well characterized in sepiolid squids, but a number of species in the family Loliginidae are also known to contain bacteriogenic light organs. Interest in loliginid light organ structure has recently arisen because of their potential as ecological niches for Vibrio harveyi, a pathogenic marine bacterium. This also implies the importance of loliginid light organs as reservoirs for V. harveyi persistence in the ocean. The present study utilized transmission and scanning electron microscopy to characterize the morphology of loliginid light organs and determined the location of bacterial symbiont cells within the tissue. It was determined that the rod-shaped loliginid symbionts lack flagella, as similarly observed in other light organ-associated bacteria. Also, the interaction of individual cells to light organ tissue is not as defined as reported for other squid-Vibrio systems. In addition, SEM observations show the presence of two pores leading to the bacterial chamber. Data presented here offer support for the hypothesis of environmental transfer of bacterial symbionts in loliginid squids.
RESUMEN
No effective vaccine or immunotherapy is presently available for patients with the hemolytic uremic syndrome (HUS) induced by Shiga-like toxin (Stx) produced by enterohaemorragic Escherichia coli (EHEC) strains, such as those belonging to the O157:H7 serotype. In this work we evaluated the performance of Bacillus subtilis strains, a harmless spore former gram-positive bacterium species, as a vaccine vehicle for the expression of Stx2B subunit (Stx2B). A recombinant B. subtilis vaccine strain expressing Stx2B under the control of a stress inducible promoter was delivered to BALB/c mice via oral, nasal or subcutaneous routes using both vegetative cells and spores. Mice immunized with vegetative cells by the oral route developed low but specific anti-Stx2B serum IgG and fecal IgA responses while mice immunized with recombinant spores developed anti-Stx2B responses only after administration via the parenteral route. Nonetheless, serum anti-Stx2B antibodies raised in mice immunized with the recombinant B. subtilis strain did not inhibit the toxic effects of the native toxin, both under in vitro and in vivo conditions, suggesting that either the quantity or the quality of the induced immune response did not support an effective neutralization of Stx2 produced by EHEC strains.
RESUMEN
OBJECTIVE: To investigate the epidemiology of hepatitis B virus (HBV) infection and viral genotypes in long distance truck drivers in Brazil. METHODS: We interviewed 641 long distance truck drivers to gather data on sociodemographics and risk factors and collected blood samples to test for HBV markers (HBsAg, anti-HBs and anti-HBc) by ELISA. HBsAg-positive samples were submitted to HBV DNA detection and genotyped. RESULTS: We found a global HBV prevalence of 18.9% (95% CI 15.9 to 22.2). Thirty truck drivers (4.7%) had only anti-HBs, suggesting that they had a previous HBV vaccination. More than 20 years in the profession, more than 15 days away from home and a history of sexual transmitted infections were independently and positivity associated with HBV markers. HBV DNA was detected in nine samples in which genotypes A (n = 5), D (n = 2) and F (n = 2) were found. CONCLUSIONS: These findings confirm that truck drivers are at high risk for hepatitis B infection and highlight the importance of having a public health policy that addresses this population and is based on the characteristics of HBV acquisition and dissemination.
Asunto(s)
Conducción de Automóvil/estadística & datos numéricos , Hepatitis B Crónica/epidemiología , Enfermedades Profesionales/epidemiología , Adulto , Brasil/epidemiología , Femenino , Antígenos de Superficie de la Hepatitis B/sangre , Virus de la Hepatitis B/aislamiento & purificación , Humanos , Masculino , Prevalencia , Análisis de Regresión , Factores SocioeconómicosRESUMEN
Blood examination by microhaematocrit and haemoculture of 459 snakes belonging to 37 species revealed 2.4% trypanosome prevalence in species of Viperidae (Crotalus durissus and Bothrops jararaca) and Colubridae (Pseudoboa nigra). Trypanosome cultures from C. durissus and P. nigra were behaviourally and morphologically indistinguishable. In addition, the growth and morphological features of a trypanosome from the sand fly Viannamyia tuberculata were similar to those of snake isolates. Cross-infection experiments revealed a lack of host restriction, as snakes of 3 species were infected with the trypanosome from C. durissus. Phylogeny based on ribosomal sequences revealed that snake trypanosomes clustered together with the sand fly trypanosome, forming a new phylogenetic lineage within Trypanosoma closest to a clade of lizard trypanosomes transmitted by sand flies. The clade of trypanosomes from snakes and lizards suggests an association between the evolutionary histories of these trypanosomes and their squamate hosts. Moreover, data strongly indicated that these trypanosomes are transmitted by sand flies. The flaws of the current taxonomy of snake trypanosomes are discussed, and the need for molecular parameters to be adopted is emphasized. To our knowledge, this is the first molecular phylogenetic study of snake trypanosomes.
Asunto(s)
Colubridae/parasitología , Filogenia , Trypanosoma/clasificación , Trypanosoma/genética , Tripanosomiasis/veterinaria , Viperidae/parasitología , Animales , Bothrops/clasificación , Bothrops/parasitología , Colubridae/clasificación , Crotalus/clasificación , Crotalus/parasitología , ADN Protozoario/análisis , ADN Ribosómico/análisis , Evolución Molecular , Interacciones Huésped-Parásitos , Datos de Secuencia Molecular , Psychodidae/parasitología , Análisis de Secuencia de ADN , Trypanosoma/fisiología , Trypanosoma/ultraestructura , Tripanosomiasis/parasitología , Tripanosomiasis/transmisión , Viperidae/clasificaciónRESUMEN
The oligopeptide-binding protein, OppA, binds and ushers oligopeptide substrates to the membrane-associated oligopeptide permease (Opp), a multi-component ABC-type transporter involved in the uptake of oligopeptides expressed by several bacterial species. In the present study, we report the cloning, purification, refolding and conformational analysis of a recombinant OppA protein derived from Xanthomonas axonopodis pv. citri (X. citri), the etiological agent of citrus canker. The oppA gene was expressed in Escherichia coli BL21 (DE3) strain under optimized inducing conditions and the recombinant protein remained largely insoluble. Solubilization was achieved following refolding of the denatured protein. Circular dichroism analysis indicated that the recombinant OppA protein preserved conformational features of orthologs expressed by other bacterial species. The refolded recombinant OppA represents a useful tool for structural and functional analyses of the X. citri protein.
Asunto(s)
Proteínas Bacterianas/aislamiento & purificación , Proteínas Bacterianas/metabolismo , Proteínas Portadoras/metabolismo , Proteínas de Transporte de Membrana/aislamiento & purificación , Proteínas de Transporte de Membrana/metabolismo , Pliegue de Proteína , Xanthomonas axonopodis/genética , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Secuencia de Bases , Proteínas Portadoras/genética , Proteínas Portadoras/aislamiento & purificación , Dicroismo Circular , Clonación Molecular , Biología Computacional/métodos , Escherichia coli/genética , Proteínas de Transporte de Membrana/química , Proteínas de Transporte de Membrana/genética , Datos de Secuencia Molecular , Operón , Plásmidos , Conformación Proteica , Desnaturalización Proteica , Renaturación de Proteína , Señales de Clasificación de Proteína , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Solubilidad , Especificidad por Sustrato , Xanthomonas axonopodis/metabolismoRESUMEN
The analysis of transcriptional temporal noise could be an interesting means to study gene expression dynamics and stochasticity in eukaryotes. To study the statistical distributions of temporal noise in the eukaryotic model system Saccharomyces cerevisiae, we analyzed microarray data corresponding to one cell cycle for 6200 genes. We found that the temporal noise follows a lognormal distribution with scale invariance at the genome, chromosomal and sub-chromosomal levels. Correlation of temporal noise with the codon adaptation index suggests that at least 70% of all protein-coding genes are a noise minimization core of the genome. Accordingly, a mathematical model of individual gene expression dynamics was proposed, using an operator theoretical approach, which reveals strict conditions for noise variability and a possible global noise minimization/optimization strategy at the genome level. Our model and data show that minimal noise does not correspond to genes obeying a strictly deterministic dynamics. The natural strategy of minimization consists in equating the mean of the absolute value of the relative variation of the expression level (alpha) with noise (eta). We hypothesize that the temporal noise pattern is an emergent property of the genome and shows how the dynamics of gene expression could be related to chromosomal organization.
Asunto(s)
Genoma Fúngico , Saccharomyces cerevisiae/genética , Transcripción Genética , Regulación Fúngica de la Expresión Génica , Redes Reguladoras de Genes , Modelos Genéticos , Modelos Estadísticos , Modelos Teóricos , Análisis de Secuencia por Matrices de Oligonucleótidos , Saccharomyces cerevisiae/metabolismo , Factores de TiempoRESUMEN
The oligopeptide-binding protein, OppA, ushers oligopeptide substrates to the membrane-associated oligopeptide permease (Opp), a multi-component ABC-type transporter involved in the uptake of oligopeptides by several bacterial species. In the present study, we report a structural model and an oligopeptide docking analysis of the OppA protein expressed by Xanthomonas axonopodis pv. citri (X. citri), the etiological agent of citrus canker. The X. citri OppA structural model showed a conserved three-dimensional structure, irrespective of the low amino acid identities with previously defined structures of Bacillus subtilis and Salmonella typhimurium orthologs. Oligopeptide docking analysis carried out with the proposed model indicated that the X. citri OppA preferentially binds tri- and tetrapeptides. The present study represents the first structural analysis of an OppA ortholog expressed by a phytopathogen and contributes to the understanding of the physiology and nutritional strategies of X. citri.
Asunto(s)
Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Proteínas Portadoras/química , Proteínas Portadoras/metabolismo , Lipoproteínas/química , Lipoproteínas/metabolismo , Oligopéptidos/metabolismo , Xanthomonas axonopodis/metabolismo , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Sitios de Unión , Proteínas Portadoras/genética , Ligandos , Lipoproteínas/genética , Modelos Moleculares , Datos de Secuencia Molecular , Oligopéptidos/química , Enfermedades de las Plantas/microbiología , Unión Proteica , Conformación Proteica , Xanthomonas axonopodis/genética , Xanthomonas axonopodis/patogenicidadRESUMEN
Bacterial ATP-binding cassette (ABC) transporters play a crucial role in the physiology and pathogenicity of different bacterial species. Components of ABC transporters have also been tested as target antigens for the development of vaccines against different bacterial species, such as those belonging to the Streptococcus genus. Streptococcus mutans is the etiological agent of dental caries, and previous studies have demonstrated that deletion of the gene encoding PstS, the substrate-binding component of the phosphate uptake system (Pst), reduced the adherence of the bacteria to abiotic surfaces. In the current study, we generated a recombinant form of the S. mutans PstS protein (rPstS) with preserved structural features, and we evaluated the induction of antibody responses in mice after sublingual mucosal immunization with a formulation containing the recombinant protein and an adjuvant derived from the heat-labile toxin from enterotoxigenic Escherichia coli strains. Mice immunized with rPstS exhibited systemic and secreted antibody responses, measured by the number of immunoglobulin A-secreting cells in draining lymph nodes. Serum antibodies raised in mice immunized with rPstS interfered with the adhesion of bacteria to the oral cavity of naive mice challenged with S. mutans. Similarly, mice actively immunized with rPstS were partially protected from oral colonization after challenge with the S. mutans NG8 strain. Therefore, our results indicate that S. mutans PstS is a potential target antigen capable of inducing specific and protective antibody responses after sublingual administration. Overall, these observations raise interesting perspectives for the development of vaccines to prevent dental caries.