Anti-Inflammatory Potential of Pteropodine in Rodents.

Pteropodine (PT) is a component of some plants with potentially useful pharmacological activities for humans. This compound has biomedical properties related to the modulation of the immune system, nervous system, and inflammatory processes. This study addresses the anti-inflammatory and antioxidant capacity of pteropodin in a murine model of arthritis and induced edema of the mouse ear. To evaluate the anti-inflammatory activity, we used the reversed passive Arthus reaction (RPAR), which includes the rat paw edema test, the rat pleurisy test, and a mouse ear edema model. The antioxidant effect of PT was evaluated by determining the myeloperoxidase enzyme activity. PT showed an anti-inflammatory effect in the different specific and non-specific tests. We found a 51, 66 and 70% inhibitory effect of 10, 20 and 40 mg/kg of PT, respectively, in the rat paw edema test. In the pleurisy assay, 40 mg/kg of PT induced a low neutrophil count (up to 36%) when compared to the negative control group, and 20 mg/kg of PT increased the content of lymphocytes by up to 28% and the pleural exudate volume decreased by 52% when compared to the negative control group, respectively. We also found an 81.4% inflammatory inhibition of the edema ear with 0.04 mg/ear of PT, and a significant myeloperoxidase enzyme inhibition by the three doses of PT tested. We conclude that PT exerted a potent anti-inflammatory effect in the acute inflammation model in rodents.

Detection and expression of SapS, a class C nonspecific acid phosphatase with O-phospho-Ltyrosine- phosphatase activity, in Staphylococcus aureus isolates from patients with chronic osteomyelitis / Detección y expresión de SapS, una fosfatasa ácida no específica de clase C con actividad de fosfatasa O-fosfotirosina, en aislamientos de Staphylococcus aureus de pacientes con osteomielitis crónica

INTRODUCTION: The identity of Staphylococcus aureus virulence factors involved in chronic osteomyelitis remains unresolved. SapS is a class C non-specific acid phosphatase and a well-known virulence factor that has been identified in S. aureus strain 154 but in protein extracts from rotting vegetables. OBJECTIVE: To identify the SapS gene and characterize the activity of SapS from S. aureus strains: 12 isolates from bone infected samples of patients treated for chronic osteomyelitis and 49 from a database with in silico analysis of complete bacterial genomes. MATERIALS AND METHODS: The SapS gene was isolated and sequenced from 12 S. aureus clinical isolates and two reference strains; 49 S. aureus strains and 11 coagulase-negative staphylococci were tested using in silico PCR. Culture media semi-purified protein extracts from the clinical strains were assayed for phosphatase activity with p-nitro-phenylphosphate, O-phospho-L-tyrosine, O-phospho-L-serine, and OphosphoL-threonine in conjunction with various phosphatase inhibitors. RESULTS: SapS was detected in the clinical and in-silico S. aureus strains, but not in the in silico coagulase-negative staphylococci strains. Sec-type I lipoprotein-type N-terminal signal peptide sequences; secreted proteins, and aspartate bipartite catalytic domains coding sequences were found in the SapS nucleotide and amino acid sequence analysis. SapS dephosphorylated with p-nitro-phenyl-phosphate and ophosphoLtyrosine were selectively resistant to tartrate and fluoride, but sensitive to vanadate and molybdate. CONCLUSION: SapS gene was found in the genome of the clinical isolates and the in silico Staphylococcus aureus strains. SapS shares biochemical similarities with known virulent bacterial, such as protein tyrosine phosphatases, suggesting it may be a virulence factor in chronic osteomyelitis.

Introducción: Se desconoce la identidad de los factores de virulencia de Staphylococcus aureus implicados en la osteomielitis crónica. Sin embargo, SapS, una fosfatasa ácida no específica de clase C, es un factor de virulencia reconocido y ya fue identificada en la cepa 154 de S. aureus, pero en extractos proteicos de vegetales podridos. Objetivo: Detectar el gen SapS y caracterizar la actividad de la fosfatasa SapS en cepas de S. aureus aisladas de pacientes con osteomielitis crónica y en las reportadas en una base de datos de análisis in silico de genomas bacterianos completos. Materiales y métodos: Se aisló y secuenció el gen SapS en los 12 aislamientos clínicos de S. aureus y en dos cepas de referencia; estas secuencias se analizaron junto con las secuencias de las cepas reportadas en la base de datos de genomas bacterianos: 49 cepas de S. aureus y 11 cepas de estafilococos negativos para coagulasa. Se evalúo la actividad de la fosfatasa SapS, presente en los extractos de los sobrenadantes de los cultivos de las cepas clínicas, mediante la hidrólisis de fosfato p-nitrofenil, O-fosfo-Ltirosina, O-fosfo-L serina y O-fosfo-L treonina junto con varios inhibidores de fosfatasas. Resultados: Se detectó el gen SapS en el genoma de las cepas clínicas y en las 49 cepas de S. aureus analizadas in silico, pero no en las 11 cepas de estafilococos negativos para coagulasa. La secuenciación de SapS reveló un péptido señal presente en el extremo N-terminal de proteínas extracelulares y los dominios bipartitos de aspartato (DDDD) en su sitio catalítico. SapS hidroliza selectivamente el fosfato p-nitrofenil y la O-fosfo-L-tirosina, pero es sensible a vanadato y molibdato. Conclusión: Se encontró SapS en el genoma de S. aureus de las cepas clínicas y de las cepas de simulación computacional. La SapS con actividad específica para la hidrólisis de la O-fosfo-L-tirosina comparte similitudes bioquímicas con las fosfatasas-tirosina bacterianas, por lo que puede formar parte de la red de factores de virulencia de la osteomielitis crónica.

Detection and expression of SapS, a class C nonspecific acid phosphatase with O-phospho-L- tyrosine-phosphatase activity, in Staphylococcus aureus isolates from patients with chronic osteomyelitis / Detección y expresión de SapS, una fosfatasa ácida no específica de clase C con actividad de fosfatasa O-fosfotirosina, en aislamientos de Staphylococcus aureus de pacientes con osteomielitis crónica

Introduction. The identity of Staphylococcus aureus virulence factors involved in chronic osteomyelitis remains unresolved. SapS is a class C non-specific acid phosphatase and a well-known virulence factor that has been identified in S. aureus strain 154 but in protein extracts from rotting vegetables. Objective. To identify the SapS gene and characterize the activity of SapS from S. aureus strains: 12 isolates from bone infected samples of patients treated for chronic osteomyelitis and 49 from a database with in silico analysis of complete bacterial genomes. Materials and methods. The SapS gene was isolated and sequenced from 12 S. aureus clinical isolates and two reference strains; 49 S. aureus strains and 11 coagulase-negative staphylococci were tested using in silico PCR. Culture media semi-purified protein extracts from the clinical strains were assayed for phosphatase activity with p-nitro-phenyl- phosphate, O-phospho-L-tyrosine, O-phospho-L-serine, and OphosphoL-threonine in conjunction with various phosphatase inhibitors. Results. SapS was detected in the clinical and in-silico S. aureus strains, but not in the in silico coagulase-negative staphylococci strains. Sec-type I lipoprotein-type N-terminal signal peptide sequences; secreted proteins, and aspartate bipartite catalytic domains coding sequences were found in the SapS nucleotide and amino acid sequence analysis. SapS dephosphorylated with p-nitro-phenyl-phosphate and ophosphoLtyrosine were selectively resistant to tartrate and fluoride, but sensitive to vanadate and molybdate. Conclusion. SapS gene was found in the genome of the clinical isolates and the in silico S. aureus strains. SapS shares biochemical similarities with known virulent bacterial, such as protein tyrosine phosphatases, suggesting it may be a virulence factor in chronic osteomyelitis.

Introducción. Se desconoce la identidad de los factores de virulencia de Staphylococcus aureus implicados en la osteomielitis crónica. Sin embargo, SapS, una fosfatasa ácida no específica de clase C, es un factor de virulencia reconocido y ya fue identificada en la cepa 154 de S. aureus, pero en extractos proteicos de vegetales podridos. Objetivo. Detectar el gen SapS y caracterizar la actividad de la fosfatasa SapS en cepas de S. aureus aisladas de pacientes con osteomielitis crónica y en las reportadas en una base de datos de análisis in silico de genomas bacterianos completos. Materiales y métodos. Se aisló y secuenció el gen SapS en los 12 aislamientos clínicos de S. aureus y en dos cepas de referencia; estas secuencias se analizaron junto con las secuencias de las cepas reportadas en la base de datos de genomas bacterianos: 49 cepas de S. aureus y 11 cepas de estafilococos negativos para coagulasa. Se evalúo la actividad de la fosfatasa SapS, presente en los extractos de los sobrenadantes de los cultivos de las cepas clínicas, mediante la hidrólisis de fosfato p-nitrofenil, O-fosfo-L- tirosina, O-fosfo-L serina y O-fosfo-L treonina junto con varios inhibidores de fosfatasas. Resultados. Se detectó el gen SapS en el genoma de las cepas clínicas y en las 49 cepas de S. aureus analizadas in silico, pero no en las 11 cepas de estafilococos negativos para coagulasa. La secuenciación de SapS reveló un péptido señal presente en el extremo N-terminal de proteínas extracelulares y los dominios bipartitos de aspartato (DDDD) en su sitio catalítico. SapS hidroliza selectivamente el fosfato p-nitrofenil y la O-fosfo-L-tirosina, pero es sensible a vanadato y molibdato. Conclusión. Se encontró SapS en el genoma de S. aureus de las cepas clínicas y de las cepas de simulación computacional. La SapS con actividad específica para la hidrólisis de la O-fosfo-L-tirosina comparte similitudes bioquímicas con las fosfatasas-tirosina bacterianas, por lo que puede formar parte de la red de factores de virulencia de la osteomielitis crónica.

Characterization of Redox Environment and Tryptophan Catabolism through Kynurenine Pathway in Military Divers' and Swimmers' Serum Samples.

Endurance and resistance exercises, alone or in combination, induce metabolic changes that affect tryptophan (Trp) catabolism. The kynurenine pathway (KP) is the main route of Trp degradation, and it is modulated by the inflammatory and redox environments. Previous studies have shown that KP metabolites work as myokines that mediate the positive systemic effects related to exercise. However, it is poorly understood how different exercise modalities and intensities impact the KP. The aim of this study was to characterize the effect of two different exercise modalities, military diving and swimming, on the KP and the redox environment. A total of 34 healthy men from the Mexican Navy were included in the study, 20 divers and 14 swimmers, who started and stayed in military training consistently during the six months of the study; 12 Mexican men without fitness training were used as the control group. Physical fitness was determined at the beginning and after 6 months of training; criteria included body composition; serum levels of Trp, kynurenine (KYN), kynurenic acid (KYNA) and 3-hydroxykynurenine (3-HK); the glutathione ratio (GSH/GSSG); and malondialdehyde (MDA).. Results showed a significant loss of body fat in both the diver and swimmer groups. Compared with the control group, divers showed a decrease in Trp and 3-HK levels, but no changes were observed in the KYN/Trp, KYNA/Trp or 3-HK/Trp ratios, while swimmers showed a decrease in KYN levels and an increase in the KYNA and 3-HK levels. Additionally, divers showed a decrease in the GSH/GSSG ratio and an increase in MDA levels, in contrast to the swimmers, who showed a decrease in MDA levels and an increase in GSH/GSSG levels. Our findings suggest a differential shift in the KP and redox environment induced by diving and swimming. Swimming promotes an antioxidant environment and a peripheral overactivation of the KP.

Production and Evaluation of an Avian IgY Immunotoxin against CD133+ for Treatment of Carcinogenic Stem Cells in Malignant Glioma: IgY Immunotoxin for the Treatment of Glioblastoma.

BACKGROUND: Glioblastoma is the most common malignant tumor of Central Nervous System. Despite the research in therapeutics, the prognosis is dismal. Malignant glioma stem cells (MGSCs) are a major cause of treatment failure and increasing tumor recurrence. In general, cancer stem cells (CSCs) express prominin-1 (CD133), considered as a potential therapeutic target. In this study, we produced an avian immunotoxin directed against the subpopulation of CD133+ CSCs within a malignant glioma. We used the avian IgY because it has various advantages as increased affinity to mammal antigens and inexpensive obtention of large amounts of specific antibodies (approximately 1 mg/per egg). The design, production, purification and use of IgY anti CD133 immunotoxin constitute an original goal of this research. METHODS: The immunodominant peptide of CD133 was designed to immunize hens; also, the extracellular domain of CD133 was cloned to probe the IgY antibodies. In parallel, a recombinant abrin A chain was produced in E. coli in order to join it to the Fc domain of the anti-CD133 IgY to conform the immunotoxin. This anti-CD133 IgY anti-tumor immunotoxin was tested in vitro and in vivo. Results. The cytotoxicity of the immunotoxin in vitro showed that IgY-abrin immunotoxin reduced 55% cell viability. After subcutaneous MGSCs implantation, the animals treated intraperitoneally or intratumorally with the IgY-abrin immunotoxin showed more than 50% decrease of tumor volume. CONCLUSION: Results showed that the IgY-abrin immunotoxin had cytotoxic activity against CD133+ MGSCs and provides a novel approach for the immunotherapy of glioblastoma.

Análisis molecular por PCR múltiple anidada para virus de la familia herpes en la parálisis facial periférica idiopática / Molecular analysis by multiplex nested PCR for herpes virus family in idiopathic peripheral facial paralysis

En el Instituto Nacional de Rehabilitación, la parálisis facial periférica idiopática (PFPI) ocupa uno de los diez primeros lugares de atención. Su etiología aún es desconocida; sin embargo, se han identificado los virus de la familia herpes (HSV) como posibles agentes causales. Objetivo: Detectar el ADN de virus HSV-1 y HVS-2, citomegalovirus (CMV) y varicela zóster (VZV) en pacientes con PFPI y correlacionar su presencia con la presentación clínica de la enfermedad. Métodos: Se extrajo el ADN de la fracción leucocitaria de 62 muestras de pacientes con PFPI. La amplificación del ADN viral se realizó por PCR múltiple anidada con oligonucleótidos específicos para cada virus. La determinación de IgG e IgMse realizó por el método de ELISA. Resultados: La PCR mostró 22 (35.5%) casos positivos para HSV-1,1(1.6%) para HSV-2, 1 (1.6%) para VZV, 3 (4.8%) para CMV. La seroprevalencia mostró que 55 (88.7%) casos presentaron niveles altos de IgG para HSV-1 y 2, 2 (3.22%) para IgG-VZV y 5 (8.1%) para IgMCMV. Tanto los casos positivos como los negativos para HSV-1 no establecieron diferencias significativas con la edad, sexo, lateralidad, síntomas, grado de parálisis facial y la temporada en la que se presentó la parálisis. Conclusión: El virus más frecuente encontrado en nuestros pacientes fue el HSV-1 lo que sugiere una fuerte asociación entre la presencia de HSV-1 y la aparición de PFPI...

[Phenotypic characterization of Staphylococcus epidermidis isolated from patients with endophthalmitis]. / Caracterización fenotípica de Staphylococcus epidermidis aislado de pacientes con endoftalmitis.

OBJECTIVE: To carry out the phenotypic characterization of Staphylococcus epidermidis isolated from endophthalmitis developed after cataract extraction and implantation of an intraocular lens. This bacteria produces a biofilm, adheres to polystyrene and host proteins such as collagen and fibronectine, significant virulence factors. METHODS: Five S. epidermidis strains were isolated from cases of endophthalmitis, they developed after crystalline extraction and implantation of an intraocular lens. We assessed if these strains adhere to polystyrene, to Type I collagen and to fibronectine and if bacteria produced biofilm. Finally, the bacterial surface proteins were obtained and analyzed using polyacrylamide gel electrophoresis. RESULTS: All five bacterial strains adhered to polystyrene, with a maximum adherence time of 105 min; they also displayed adherence to fibronectine but only two to collagen. Only two strains were weak biofilm producers. We identified proteins that by molecular weight are similar to those identified in the literature as proteins binding to biomaterials. CONCLUSIONS: As the strains that we studied were not biofilm-forming they should be considered as non-pathogenic. Nevertheless, they meet the initial criteria of pathogenicity and adherence, aside from being isolated from an intraocular infectious process and being able to provoke endophtalmitis when inoculated in rabbit eyes.

Caracterización fenotípica de Staphylococcus epidermidis aislado de pacientes con endoftalmitis / Phenotypic characterization of Staphylococcus epidermidis isolated from patients with endophthalmitis

Objetivo: Conocer las características fenotípicas de Staphylococcus epidermidis aislado de endoftalmitis relacionadas con el implante de lente intraocular de metilmetacrilato, su capacidad para formar biofilm y adherencia a proteínas de matriz extracelular y poliestireno. Métodos: Se estudiaron cinco cepas de Staphylococcus epidermidis aisladas de enfermos con endoftalmitis posterior la extracción del cristalino con implante de lente intraocular. Se investigó si estas cepas se adhieren a poliestireno, a colágena tipo I y a fibronectina, así como si las bacterias eran formadoras de biofilm. Al final se extrajeron las proteínas de superficie de las bacterias y se analizaron por electroforesis en gel de poliacrilamida. Resultados: Se encontró que las cinco cepas se unieron al poliestireno y que lo hicieron con mayor eficacia en la fase de crecimiento exponencial, con máxima adherencia a los 105 minutos; las cinco cepas se adhirieron a fibronectina y solo dos (CV y EN) a colágena. Dos cepas (CV y EN) fueron débiles formadoras de biofilm. Se identificaron proteínas que por peso molecular corresponden con las informadas en la literatura como proteínas de unión a biomateriales. Conclusiones: Las cepas estudiadas al no ser formadoras de biofilm tendrían que ser consideradas no patógenas, pero cumplen con el paso inicial de la patogenicidad, la adherencia, además de que fueron aisladas de un proceso infeccioso intraocular y produjeron endoftalmitis cuando fueron inoculadas en ojos de conejo.

OBJECTIVE: To carry out the phenotypic characterization of Staphylococcus epidermidis isolated from endophthalmitis developed after cataract extraction and implantation of an intraocular lens. This bacteria produces a biofilm, adheres to polystyrene and host proteins such as collagen and fibronectine, significant virulence factors. METHODS: Five S. epidermidis strains were isolated from cases of endophthalmitis, they developed after crystalline extraction and implantation of an intraocular lens. We assessed if these strains adhere to polystyrene, to Type I collagen and to fibronectine and if bacteria produced biofilm. Finally, the bacterial surface proteins were obtained and analyzed using polyacrylamide gel electrophoresis. RESULTS: All five bacterial strains adhered to polystyrene, with a maximum adherence time of 105 min; they also displayed adherence to fibronectine but only two to collagen. Only two strains were weak biofilm producers. We identified proteins that by molecular weight are similar to those identified in the literature as proteins binding to biomaterials. CONCLUSIONS: As the strains that we studied were not biofilm-forming they should be considered as non-pathogenic. Nevertheless, they meet the initial criteria of pathogenicity and adherence, aside from being isolated from an intraocular infectious process and being able to provoke endophtalmitis when inoculated in rabbit eyes.