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1.
J Dairy Sci ; 106(4): 2772-2783, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36870844

RESUMEN

We examined whether distinct staphylococcal and mammaliicoccal species and strains trigger B- and T-lymphocyte proliferation and interleukin (IL)-17A and interferon (IFN)-γ production by peripheral blood mononuclear cells in nulliparous, primiparous, and multiparous dairy cows. Flow cytometry was used to measure lymphocyte proliferation with the Ki67 antibody, and specific monoclonal antibodies were used to identify CD3, CD4, and CD8 T lymphocyte and CD21 B lymphocyte populations. The supernatant of the peripheral blood mononuclear cell culture was used to measure IL-17A and IFN-γ production. Two distinct, inactivated strains of bovine-associated Staphylococcus aureus [one causing a persistent intramammary infection (IMI) and the other from the nose], 2 inactivated Staphylococcus chromogenes strains [one causing an IMI and the other from a teat apex), as well as an inactivated Mammaliicoccus fleurettii strain originating from sawdust from a dairy farm, and the mitogens concanavalin A and phytohemagglutinin M-form (both specifically to measure lymphocyte proliferation) were studied. In contrast to the "commensal" Staph. aureus strain originating from the nose, the Staph. aureus strain causing a persistent IMI triggered proliferation of CD4+ and CD8+ subpopulations of T lymphocytes. The M. fleurettii strain and the 2 Staph. chromogenes strains had no effect on T- or B-cell proliferation. Furthermore, both Staph. aureus and Staph. chromogenes strains causing persistent IMI significantly increased IL-17A and IFN-γ production by peripheral blood mononuclear cells. Overall, multiparous cows tended to have a higher B-lymphocyte and a lower T-lymphocyte proliferative response than primiparous and nulliparous cows. Peripheral blood mononuclear cells of multiparous cows also produced significantly more IL-17A and IFN-γ. In contrast to concanavalin A, phytohemagglutinin M-form selectively stimulated T-cell proliferation.


Asunto(s)
Enfermedades de los Bovinos , Mastitis Bovina , Infecciones Estafilocócicas , Femenino , Bovinos , Animales , Fitohemaglutininas , Interleucina-17 , Concanavalina A , Leucocitos Mononucleares , Staphylococcus aureus/fisiología , Infecciones Estafilocócicas/veterinaria , Anticuerpos Monoclonales , Proliferación Celular , Leche
2.
BMC Vet Res ; 17(1): 282, 2021 Aug 25.
Artículo en Inglés | MEDLINE | ID: mdl-34433467

RESUMEN

BACKGROUNDS: The present study explored the viability of bovine milk macrophages, their intracellular production of reactive oxygen and nitrogen species (RONS), and their phagocytosis of Staphylococcus aureus, as well as the profile of lymphocytes, from healthy udder quarters and udder quarters infected by Corynebacterium bovis. The study included 28 healthy udder quarters from 12 dairy cows and 20 udder quarters infected by C. bovis from 10 dairy cows. The percentages of macrophages and lymphocytes were identified by flow cytometry using monoclonal antibodies. Macrophage viability, RONS production, and S. aureus phagocytosis were evaluated by flow cytometry. RESULTS: Milk samples from quarters infected with C. bovis showed a lower percentage of macrophages but an increased number of milk macrophages per mL and a higher percentage of macrophages that produced intracellular RONS and phagocytosed S. aureus. No effect of C. bovis infection on macrophage viability was found. Udder quarters infected by C. bovis showed a higher percentage of T cells and CD4+ T lymphocytes, but no effect was found on the percentage of CD8+ CD4- T, CD8- CD4- T, or B lymphocytes. CONCLUSIONS: Thus, our results corroborate, at least in part, the finding that intramammary infections by C. bovis may offer protection against intramammary infections by major pathogens.


Asunto(s)
Macrófagos/fisiología , Mastitis Bovina/microbiología , Leche/citología , Animales , Bovinos , Corynebacterium , Femenino , Linfocitos , Mastitis Bovina/patología , Fagocitosis , Especies de Nitrógeno Reactivo , Especies Reactivas de Oxígeno , Staphylococcus aureus
3.
Cell Microbiol ; 16(10): 1549-64, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24824158

RESUMEN

The last step of Leishmania intracellular life cycle is the egress of amastigotes from the host cell and their uptake by adjacent cells. Using multidimensional live imaging of long-term-infected macrophage cultures we observed that Leishmania amazonensis amastigotes were transferred from cell to cell when the donor host macrophage delivers warning signs of imminent apoptosis. They were extruded from the macrophage within zeiotic structures (membrane blebs, an apoptotic feature) rich in phagolysosomal membrane components. The extrusions containing amastigotes were selectively internalized by vicinal macrophages and the rescued amastigotes remain viable in recipient macrophages. Host cell apoptosis induced by micro-irradiation of infected macrophage nuclei promoted amastigotes extrusion, which were rescued by non-irradiated vicinal macrophages. Using amastigotes isolated from LAMP1/LAMP2 knockout fibroblasts, we observed that the presence of these lysosomal components on amastigotes increases interleukin 10 production. Enclosed within host cell membranes, amastigotes can be transferred from cell to cell without full exposure to the extracellular milieu, what represents an important strategy developed by the parasite to evade host immune system.


Asunto(s)
Leishmania/patogenicidad , Leishmaniasis/transmisión , Proteínas de Membrana de los Lisosomas/metabolismo , Proteína 2 de la Membrana Asociada a los Lisosomas/metabolismo , Macrófagos/parasitología , Animales , Apoptosis , Línea Celular , Membrana Celular/parasitología , Fibroblastos , Interleucina-10/biosíntesis , Leishmaniasis/patología , Proteína 2 de la Membrana Asociada a los Lisosomas/genética , Proteínas de Membrana de los Lisosomas/genética , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL
4.
RSC Adv ; 13(8): 5070-5080, 2023 Feb 06.
Artículo en Inglés | MEDLINE | ID: mdl-36762084

RESUMEN

Perovskite solar cells (PSCs) are an evolving photovoltaic field with the potential to disrupt the established silicon solar cell market. However, the presence of many transport barriers and defect trap states at the interfaces and grain boundaries has negative effects on PSCs; it decreases their efficiency and stability. The purpose of this work was to investigate the effects on efficiency and stability achieved by quaternary theophylline additives in MAPbI3 PSCs with the structure FTO/TiO2/perovskite/spiro-OMeTAD/Ag. The X-ray photoelectron spectroscopy (XPS) and theoretical calculation strategies were applied to study the additive's interaction in the layer. The tetrafluoroborinated additive results in an increase in device current density (J SC) (23.99 mA cm-1), fill factor (FF) (65.7%), and open-circuit voltage (V OC) (0.95 V), leading to significant improvement of the power conversion efficiency (PCE) to 15.04% compared to control devices (13.6%). Notably, films exposed to controlled humidity of 30% using the tetrafluoroborinated additive maintained their stability for more than 600 hours (h), while the control films were stable for less than 240 hours (h).

5.
Vet Microbiol ; 283: 109792, 2023 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-37269712

RESUMEN

The discovery and tracking of antimicrobial resistance genes are essential for understanding the evolution of bacterial resistance and restraining its dispersion. Mammaliicoccus sciuri (formerly Staphylococcus sciuri) is the most probable evolutionary repository of the mecA gene, that later disseminated to S. aureus. In this study, we describe the first double mecA/mecC homologue-positive non-aureus staphylococci and mammaliicocci (NASM) from the American continent, also representing the first report of mecC-positive NASM in Brazil. Two clonally related methicillin-resistant M. sciuri strains co-carrying mecA and mecC genes were isolated from the teat skin swab and milk sample collected from an ewe's left udder half. Both M. sciuri strains belonged to the sequence type (ST) 71. Besides mecA and mecC genes, the M. sciuri strains carried broad resistomes for clinically important antimicrobial agents, including ß-lactams, tetracyclines, lincosamide, streptogramin, streptomycin, and aminoglycosides. Virulome analysis showed the presence of the clumping factor B (clfB), ATP-dependent protease ClpP (ClpP) and serine-aspartate repeat proteins (sdrC and sdrE) virulence-associated genes. Phylogenomic analysis revealed that these M. sciuri strains are part of a globally disseminated branch, associated with farm and companion animals and even with food. Our findings suggest that M. sciuri is likely to emerge as a pathogen of global interest, carrying a broad repertoire of antimicrobial resistance genes with a remarkable co-presence of mecA and mecC genes. Finally, we strongly encourage to monitor M. sciuri under the One Health umbrella since this bacterial species is spreading at the human-animal-environment interface.


Asunto(s)
Staphylococcus aureus Resistente a Meticilina , Enfermedades de las Ovejas , Infecciones Estafilocócicas , Femenino , Ovinos , Animales , Humanos , Staphylococcus aureus/genética , Ganado , Brasil/epidemiología , Proteínas Bacterianas/genética , Antibacterianos/farmacología , Infecciones Estafilocócicas/epidemiología , Infecciones Estafilocócicas/veterinaria , Infecciones Estafilocócicas/microbiología , Pruebas de Sensibilidad Microbiana/veterinaria
6.
Front Microbiol ; 14: 1283738, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38173670

RESUMEN

Guinea pigs have historically been used as a food source and are also an important model for studying the human intestines. Fasting is the act of temporarily stopping the intake of food. This process can alter the microbiota of various animals. This study is the first to investigate the impact of fasting on the cecum microbiome of three guinea pig breeds. We investigated the impact of fasting on the microbiome population structure in the cecum of three guinea pig breeds. This was done by sequencing and analyzing the V4 hypervariable region of the 16S rRNA gene in bacterial communities found in cecum mucosa samples. To achieve this, we established two treatment groups (fasting and fed), for each of the three guinea pig breeds: Andina, Inti, and Peru. The study involved twenty-eight guinea pigs, which were divided into the following groups: Andina-fed (five), Andina-fasting (five), Inti-fed (four), Inti-fasting (five), Peru-fed (five), and Peru-fasting (four). The results indicated a significant difference in beta diversity between the treatment groups for the Peru breed (P-value = 0.049), but not for the treatment groups of the Andina and Inti breeds. The dominant phyla across all groups were Firmicutes and Bacteroidetes. We observed variations in the abundance of different taxa in the cecum microbiota when comparing the treatment groups for each breed. Additionally, there was a higher number of unique taxa observed in the fasting groups compared to the fed groups. We discovered that the genus Victivallis was the only one present in all fasting groups across all breeds. Despite the findings, the resilience of the gut microbiome was not challenged in all three breeds, which can lead to disruptive changes that may affect the overall maintenance of the cecum microbiome. Based on the observed differences in the treatment groups of the Peru breed, it can be suggested that fasting has a greater impact on this particular breed.

7.
Vet Immunol Immunopathol ; 253: 110508, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36327943

RESUMEN

Staphylococcus aureus mastitis constitutes a serious threat to dairy cows. The reasons why available vaccines are not fully effective remain poorly understood; thus, in the present study, we investigated CD4+ and CD8+ T lymphocyte proliferation in dairy cows vaccinated with a polyvalent mastitis vaccine that had distinct precedent Staphylococcus aureus mastitis. We studied 17 S. aureus-infected dairy cows (11 vaccinated and six unvaccinated) and eight vaccinated healthy dairy cows with no previous S. aureus mastitis infections. Flow cytometry was used to assess lymphocyte proliferation using an anti-Ki67 antibody, and monoclonal antibodies were used to identify T cell subsets. S. aureus-infected cows exhibited reduced overall lymphocyte proliferation, including CD4+ T lymphocyte proliferation, and memory lymphocyte proliferation in response to S. aureus isolate stimulus. Immunization did not influence the expansion of blood lymphocyte populations. Furthermore, CD8+ T cells, memory CD8+ T lymphocytes, and effector memory CD8+ T lymphocytes displayed reduced proliferation 21 days after the third vaccine dose compared with before vaccination at time zero. The present data demonstrates an overall negative regulation of the T-cell response suggesting its detrimental impact leading to the persistence of S. aureus intramammary infections. Furthermore, the lack of vaccination effect on T-cell mediated immunity (e.g., proliferation) may be related to poor vaccine efficacy.


Asunto(s)
Mastitis Bovina , Infecciones Estafilocócicas , Vacunación , Animales , Bovinos , Femenino , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/inmunología , Proliferación Celular , Mastitis Bovina/inmunología , Mastitis Bovina/prevención & control , Leche , Infecciones Estafilocócicas/prevención & control , Infecciones Estafilocócicas/veterinaria , Staphylococcus aureus , Vacunas Bacterianas/inmunología , Vacunación/veterinaria
8.
Antibiotics (Basel) ; 11(12)2022 Dec 16.
Artículo en Inglés | MEDLINE | ID: mdl-36551488

RESUMEN

BACKGROUND: Staphylococcus aureus is one of the most frequently major mastitis pathogens that cause clinical and subclinical mastitis worldwide. Current antimicrobial treatments are usually ineffective, and the commercially available vaccines lack proven effectiveness. The immunological response elicited by the recombinant S. aureus-cure-associated proteins phosphoglycerate kinase (PGK), enolase (ENO), and elongation factor-G (EF-G) in combination with the granulocyte-macrophage colony-stimulating factor (GM-CSF) DNA vaccination was studied in this work. METHODS: Here, twenty-three C57BL/6 mice were divided into four groups and vaccinated with: G1: none (control); G2: GM-CSF DNA plasmid DNA vaccine; G3: the combination of EF-G+ENO+PGK; and G4: the combinations of EF-G+ENO+PGK proteins plus GM-CSF plasmid DNA vaccine. After 44 days, spleen cells were collected for immunophenotyping and lymphocyte proliferation evaluation by flow cytometry upon S. aureus stimulus. RESULTS: Immunization with the three S. aureus recombinant proteins alone resulted in a higher percentage of IL-17A+ cells among CD8+ T central memory cells, as well as the highest intensity of IL-17A production by overall lymphocytes indicating that the contribution of the combined lymphocyte populations is crucial to sustaining a type 3 cell immunity environment. CONCLUSION: The immunization with three S. aureus-cure-associated recombinant proteins triggered type 3 immunity, which is a highly interesting path to pursue an effective bovine S. aureus mastitis vaccine.

9.
Chem Soc Rev ; 39(8): 3210-39, 2010 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-20577662

RESUMEN

The development of high-temperature PEM fuel cells (working at 150-200 degrees C) is pursued worldwide in order to solve some of the problems of current cells based on Nafion (CO tolerance, improved kinetics, water management, etc.). Polybenzimidazole membranes nanoimpregnated with phosphoric acid have been studied as electrolytes in PEMFCs for more than a decade. Commercially available polybenzimidazole (PBI) has been the most extensively studied and used for this application in membranes doped with all sorts of strong inorganic acids. In addition to this well-known polymer we also review here studies on ABPBI and other polybenzimidazole type membranes. More recently, several copolymers and related derivatives have attracted many researchers' attention, adding variety to the field. Furthermore, besides phosphoric acid, many other strong inorganic acids, as well as alkaline electrolytes have been used to impregnate benzimidazole membranes and are analyzed here. Finally, we also review different hybrid materials based on polybenzimidazoles and several inorganic proton conductors such as heteropoly acids, as well as sulfonated derivatives of the polymers, all of which contribute to a quickly-developing field with many blooming results and useful potential which are the subject of this critical review (317 references).

10.
Vaccines (Basel) ; 9(8)2021 Aug 13.
Artículo en Inglés | MEDLINE | ID: mdl-34452024

RESUMEN

Staphylococcus aureus mastitis remains a major challenge for dairy farming. Here, 24 mice were immunized and divided into four groups: G1: control; G2: Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) DNA vaccine; G3: F0F1 ATP synthase subunit α (SAS), succinyl-diaminopimelate (SDD), and cysteinyl-tRNA synthetase (CTS) recombinant proteins; and G4: SAS+SDD+CTS plus GM-CSF DNA vaccine. The lymphocyte subpopulations, and the intracellular interleukin-17A (IL-17A) and interferon-γ production in the draining lymph node cells were immunophenotyped by flow cytometry. The immunophenotyping and lymphocyte proliferation was determined in spleen cells cultured with and without S. aureus stimulus. Immunization with S. aureus recombinant proteins generated memory cells in draining lymph nodes. Immunization with the three recombinant proteins plus GM-CSF DNA led to an increase in the percentage of IL-17A+ cells among overall CD44+ (memory), T CD4+, CD4+ T CD44+ CD27-, γδ TCR, γδ TCR+ CD44+ CD27+, and TCRVγ4+ cells. Vaccination with S. aureus recombinant proteins associated with GM-CSF DNA vaccine downregulated TH2 immunity. Immunization with the three recombinant proteins plus the GM-CSF DNA led to a proliferation of overall memory T, CD4+, and CD4+ TEM cells upon S. aureus stimulus. This approach fostered type 3 immunity, suggesting the development of a protective immune response against S. aureus.

11.
Vet Immunol Immunopathol ; 196: 53-59, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29695326

RESUMEN

The present study aimed to validate the use of R-phycoerythrin (R-PE)-labeled Mannheimia haemolytica to simultaneously stimulate phagocytosis and intracellular production of reactive oxygen species (ROS) by blood phagocytes in bronchoalveolar lavage (BAL) fluid. Initially, R-PE-labeled M. haemolytica was inactivated using a water bath at 60 °C for 60 min. Afterwards, R-PE labelling of bacteria was confirmed by flow cytometry. The geometric mean fluorescence intensity of R-PE-labeled bacteria (FL2 detector, 585 ±â€¯42 nm) was analyzed by flow cytometry and was 41.5-fold higher than the respective unlabeled controls, confirming the success of bacterial conjugation to R-PE. Phagocytosis and intracellular production of ROS by blood neutrophils and monocytes, and by BAL CD14+ macrophages, in 12 healthy 6-month-old male calves were then performed using R-PE-labeled bacteria and 2',7'-dichlorofluorescein diacetate (DCFH-DA) as probes. Confocal microscopy was used to confirm phagocytosis of R-PE-labeled M. haemolytica by phagocytes and to exclude erroneous measurements of bacteria adhering to the leukocyte membrane. The present study showed that there is no difference in the ROS production without stimulus and in the presence of M. haemolytica by peripheral blood neutrophils and monocytes, in contrast to the increased ROS production by local alveolar macrophages upon stimulation by M. haemolytica. This emphasizes the importance of alveolar macrophages in the maintenance of homeostasis and health of the respiratory system, which can be supported during the inflammatory process by the rapid recruitment of neutrophils with high microbicidal and phagocytic capacity. The method described here provides an easy and feasible tool to measure phagocytosis and intracellular ROS production by phagocytes, especially when commonly used probes for intracellular ROS production were used, such as DCFH-DA and dihydrorhodamine 123.


Asunto(s)
Líquido del Lavado Bronquioalveolar/citología , Macrófagos/metabolismo , Mannheimia haemolytica/metabolismo , Monocitos/metabolismo , Neutrófilos/metabolismo , Fagocitosis , Ficoeritrina/uso terapéutico , Especies Reactivas de Oxígeno/metabolismo , Animales , Bovinos , Citometría de Flujo/veterinaria , Macrófagos/química , Macrófagos/inmunología , Macrófagos Alveolares/química , Macrófagos Alveolares/inmunología , Macrófagos Alveolares/metabolismo , Masculino , Mannheimia haemolytica/inmunología , Microscopía Confocal/veterinaria , Monocitos/química , Monocitos/inmunología , Neutrófilos/química , Neutrófilos/inmunología , Especies Reactivas de Oxígeno/análisis
12.
Blood Transfus ; 16(1): 17-25, 2018 01.
Artículo en Inglés | MEDLINE | ID: mdl-27893347

RESUMEN

BACKGROUND: Transfusion-transmitted malaria due to asymptomatic Plasmodium infections is a challenge for blood banks. There is a lack of data on the prevalence of asymptomatic infected blood donors and the incidence of transfusion-transmitted malaria in low endemicity areas worldwide. We estimated the frequency of blood donors harbouring Plasmodium in an area in which asymptomatic infections have been reported. MATERIAL AND METHODS: To estimate the frequency of blood donors harbouring Plasmodium we used microscopy and molecular tools. Serological tests were applied to measure the exposure of candidates to Plasmodium antigens. Venous blood was collected from 91 candidates attending the "Pró-Sangue" Blood Centre Foundation in São Paulo, who lived in the municipality of Juquitiba, São Paulo, Brazil, where sporadic autochthonous cases of malaria have been described. Blood samples were used for parasitological, molecular and serological studies. RESULTS: Among the 91 samples examined, rare Plasmodium forms were observed in two donors. Genus real-time polymerase chain reaction analysis demonstrated Plasmodium amplification in three candidates and species-specific nested polymerase chain reaction identified P. malariae in two. ELISA-IgG was reactive in 42.9% of samples for P. vivax (Pv-MSP119) and in 6.6% for P. falciparum (Pf-Zw). ELISA-IgM was reactive in 2.2% of samples for P. vivax and in 4.4% for P. falciparum. An indirect immunofluorescence assay was reactive for P. malariae in 15.4% of cases. DISCUSSION: Reservoirs of Plasmodium represent a challenge for blood banks, since studies have shown that high levels of submicroscopic infections can occur in low transmission areas. The risk of transfusion-transmitted malaria presented here points to the need to conduct molecular investigations of candidate donors with any positive malarial antibody test.


Asunto(s)
Antígenos de Protozoos/sangre , Donantes de Sangre , Selección de Donante/métodos , Malaria/sangre , Plasmodium , Femenino , Humanos , Malaria/transmisión , Masculino
13.
J Pharm Pharmacogn Res ; 6(3): 179-190, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30739984

RESUMEN

CONTEXT: The recombinant human erythropoietin (rHuEPO) stimulates the erythropoiesis process. Because this glycoprotein has a short half-life, it needs to be administrated two to three times a week. One of the technics to solve this issue is the PEGgilation. AIMS: To evaluate the pharmacokinetics (PK) and pharmacodynamics of two new branched PEGylated erythropoietins (i.e., an asymmetric 32 kDa-PEG2-rHuEPO and a symmetric 40 kDa-PEG2-rHuEPO molecule) compared to non-PEGylated ior®EPOCIM and MIRCERA®. METHODS: Serum concentrations of both PEGylated and non-PEGylated erythropoietins were measured at various time points in order to determine PK parameters using non-compartmental analysis approach. The reticulocyte (%), erythrocyte count and hemoglobin levels were ascertained in order to compare the effect of these molecules after administrating a single intravenous dose (10 µg/kg) of each product in male New Zealand rabbits. RESULTS: Both branched PEGylated erythropoietin forms exhibited half-lives that were significantly longer than ior®EPOCIM (p<0.05), but not statistically different to MIRCERA®. The mean elimination half-life increased from 4 h (ior®EPOCIM) to 131 h for the 32 kDa-PEG2-rHuEPO and 119 h for the 40 kDa-PEG2-rHuEPO. Conversely, MIRCERA® exhibits a half-life of 64 h. Both PEGylated erythropoietin products significantly enhanced the stimulating effect on reticulocytes and erythrocytes formation, as well as on hemoglobin levels, when compared to ior®EPOCIM treatment up to 42 days post-dose. CONCLUSIONS: The PEGylation strategy employed in this study is an effective method to modify the pharmacokinetics and pharmacodynamics of rHuEPO molecule achieving higher half-lives and, therefore, longer in vivo bioactivity. Both of the branched PEGylated-EPO forms tested are promising candidates for human testing.


CONTEXTO: La eritropoyetina humana recombinante (rHuEPO) estimula la formación de eritrocitos en la medula ósea. Esta glicoproteína terapéutica presenta rápida eliminación en el organismo. Una de las estrategias tecnológicas para resolver esta problemática es la PEGgilación. OBJETIVOS: Evaluar la farmacocinética y la farmacodinámica de dos nuevas eritropoyetinas PEGiladas ramificadas (una asimétrica de 32 kDa-PEG2-rHuEPO y otra simétrica de 40 kDa-PEG2-rHuEPO) en comparación con ior®EPOCIM y el producto de referencia MIRCERA®. MÉTODOS: Se midieron las concentraciones séricas de eritropoyetina PEGilada y no PEGilada, a diferentes tiempos, para determinar los parámetros farmacocinéticos usando el método de análisis no compartimental. Se determinaron los % de reticulocitos, los niveles de eritrocitos y hemoglobina para comparar el efecto de estas moléculas después de administrar a dosis única 10 µg/kg por vía intravenosa en conejos Nueva Zelanda machos. RESULTADOS: Las eritropoyetinas PEGiladas ramificadas presentaron semividas significativamente superiores a ior®EPOCIM (p<0.05), pero no fueron estadísticamente diferentes a MIRCERA®. El t1/2 aumentó de 4 h (ior®EPOCIM) a 131 h para la 32 kDa-PEG2-rHuEPO y 119 h para la 40 kDa-PEG2-rHuEPO, respectivamente. Ambas eritropoyetinas PEGiladas mejoraron significativamente el efecto estimulante sobre la formación de reticulocitos y eritrocitos, así como los niveles de hemoglobina, en comparación con ior®EPOCIM hasta 42 días después de la dosis. CONCLUSIONES: La estrategia de PEGilación, empleada en este estudio, es un método efectivo para modificar la farmacocinética y farmacodinamia de moléculas de eritropoyetinas. Esta tecnología permitió aumentar la semivida de estas moléculas, así como prolongar su bioactividad in vivo. Ambas formas ramificadas de rHuEPO PEGiladas son candidatos prometedores para su uso clínico.

14.
Ultrason Sonochem ; 36: 95-100, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28069244

RESUMEN

In this work, an easy, fast and environmentally friendly method to obtain Bi2S3 nanostructures with sphere-like morphology is introduced. The promising material was successfully synthesized by a sonochemical route in 20% 1-ethyl-3-methylimidazolium ethyl sulfate [EMIM][EtSO4] ionic liquid solution (IL). Morphological studies by electron microscopy (SEM and TEM) show that the use of IL in the synthesis of Bi2S3 favors the formation of nanocrystals non-agglomerated. Micro Raman and energy dispersive X-ray spectroscopy (EDXS) were used to determine the composition and purity of the synthesized material. X-ray powder diffraction (XRD) and selective area electron diffraction (SAED) revealed that ultrasonic radiation accelerated the crystallization of Bi2S3 into orthorhombic bismuthinite structure. The band gap calculated from the diffuse reflectance spectra (DRS) was found to be 1.5eV.

16.
Vet Immunol Immunopathol ; 151(1-2): 163-7, 2013 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-23177628

RESUMEN

Visceral leishmaniasis (VL) caused in the New World by Leishmania (Leishmania) infantum chagasi has dog as important peridomestic reservoir in its transmission cycle. Since VL in infected animals is similar to human VL, its study is interesting for human pathology. During Leishmania infection, insulin-like growth factor-I (IGF-I) plays a role in the host-parasite interaction, favoring parasite growth, particularly acting directly on Leishmania. We evaluated IGF-I mRNA expression in different organs/tissues, which was differently modulated in dogs naturally infected by L. (L.) infantum chagasi. We also evaluated the hepatic IGF-I mRNA and serum IGF-I levels in infected dogs. Hepatic mRNA IGF-I expression was higher in the infected dogs than in control animals. However, the serum levels of IGF-I, which are related to the production of this factor in the liver, were reduced in the infected dogs compared with the non-infected controls. Thus, we suggest interference in post-transcriptional processing in IGF-I production in active visceral leishmaniasis.


Asunto(s)
Enfermedades de los Perros/genética , Enfermedades de los Perros/inmunología , Factor I del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Leishmania infantum , Leishmaniasis Visceral/veterinaria , Animales , Secuencia de Bases , Estudios de Casos y Controles , Reservorios de Enfermedades/parasitología , Reservorios de Enfermedades/veterinaria , Enfermedades de los Perros/parasitología , Perros , Femenino , Expresión Génica , Humanos , Leishmaniasis Visceral/genética , Leishmaniasis Visceral/inmunología , Leishmaniasis Visceral/parasitología , Hígado/inmunología , Masculino , Procesamiento Proteico-Postraduccional , ARN Mensajero/genética , ARN Mensajero/metabolismo
17.
J Inorg Biochem ; 123: 11-7, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23499789

RESUMEN

Amino-functionalized luminescent silica particles were investigated for use in immunoassays. The particles were prepared by the Stöber method where the [Eu(TTA)3(H2O)2] complex (TTA: 3-thenoyltrifluoroacetonate) was incorporated into silica particles during the hydrolysis and condensation of TEOS: tetraethylorthosilicate. Then, the amino groups were introduced in the particle surface using APTS: 3-aminopropyltriethoxisilane. The resulting particles were characterized by scanning electron microscopy (SEM), X ray diffraction (XRD) and photoluminescence spectroscopy. In order to demonstrate the viability of the use of luminescent particles as optical markers, an enzyme-substrate reaction was performed using HRP: horseradish peroxidase. It was possible to verify the binding of HRP-oxidized LDL (low density lipoprotein) and anti-oxLDL antibody-luminescent silica particles through the evaluation of the presence of HRP. The bioassay data open a broad field for the development of protein-tagged luminescent particles for use in biomedical sciences.


Asunto(s)
Europio/química , Luminiscencia , Dióxido de Silicio/química , Bioensayo/métodos
18.
Vet Immunol Immunopathol ; 143(1-2): 162-6, 2011 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-21665295

RESUMEN

Bovine leukemia virus (BLV) is among the most widespread livestock pathogens in many countries. Despite advances in understanding the pathogenesis of this disease, little is known about the involvement of oxidative stress. Therefore, this study examined the antioxidant status and the markers of oxidative stress in BLV-infected dairy cows. BLV infection was associated with an increase in triacylglycerol levels, a decrease in glutathione peroxidase (GSH-Px) activity and a tendency toward lower superoxide dismutase activity in the infected animals. No significant difference was observed in other markers of oxidative stress (i.e., conjugated dienes, hydroperoxides and malondialdehyde) in the infected animals compared to controls. A novel method for the analysis of oxidative stress, Z-scan based on the measurement of the mean-value of θ in low density lipoprotein indicated that the infected animals had low-density lipoprotein particles that were slightly less modified than those from the healthy group. Thus, we conclude that BLV infection is associated with a selective decrease in GSH-Px activity without any alteration in the common plasma markers of oxidative stress.


Asunto(s)
Antioxidantes/metabolismo , Leucosis Bovina Enzoótica/sangre , Estrés Oxidativo , Animales , Biomarcadores/sangre , Bovinos , Femenino , Glutatión Peroxidasa/sangre , Lípidos/sangre , Lipoproteínas LDL/sangre
19.
J Phys Chem B ; 114(12): 4271-5, 2010 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-20205443

RESUMEN

In this work, we report the physicochemical properties of a variety of aliphatic phosphonium iodide (API) salts, including thermal degradation. Also, we compared the conductivity, viscosity, and fragility behavior of APIs to similar molten systems and related these properties to their structure. Our investigation has found that APIs have an intermediate fragility behavior. We plotted the conductivity and viscosity data of APIs according to Walden's rule and found that they can be classified as an associated ionic liquid (AIL), an intermediate between a true ionic liquid and a molecular species. Lastly, we correlated the structure and behavior of APIs and similar polarizable anions, such as the diacetamide anion.

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