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1.
J Hazard Mater ; 433: 128764, 2022 07 05.
Article in English | MEDLINE | ID: mdl-35390620

ABSTRACT

Lindane and other 1,2,3,4,5,6-hexachlorocyclohexane (HCH) isomers are persistent organic pollutants highly hydrophobic, which hampers their availability and biodegradation. This work aimed at (i) investigating genes encoding enzymes involved in HCH degradation in the bacterium Sphingobium sp. D4, (ii) selecting strains, from a collection of environmental isolates, able to mobilize HCHs from contaminated soil, and (iii) analysing the biodegradation of HCHs by strain D4 in co-culture with HCH-mobilizing strains or when cultivated with root exudates. Fragments of the same size and similar sequence to linA and linB genes were successfully amplified. Two isolates, Streptomyces sp. M7 and Rhodococcus erythropolis ET54b able to produce emulsifiers and to mobilize HCH isomers from soil were selected. Biodegradation of HCH isomers by strain D4 was enhanced when co-inoculated with HCH mobilizing strains or when cultivated with root exudates. The degrader strain D4 was able to decompose very efficiently HCHs isomers, reducing their concentration in soil slurries by more than 95% (from an average initial amount of 50 ± 8 mg HCH kg-1 soil) in 9 days. The combination of HCH-degrading and HCH-mobilizing strains can be considered a promising inoculum for future soil bioremediation studies using bioaugmentation techniques or in combination with plants in rhizodegradation assays.


Subject(s)
Soil Pollutants , Sphingomonadaceae , Biodegradation, Environmental , Coculture Techniques , Exudates and Transudates/chemistry , Exudates and Transudates/metabolism , Hexachlorocyclohexane/chemistry , Soil/chemistry , Soil Pollutants/metabolism , Sphingomonadaceae/genetics , Sphingomonadaceae/metabolism
2.
Vet Parasitol ; 292: 109399, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33711619

ABSTRACT

Nematodes develop resistance to the most common commercially available drugs. The aim of this study was to identify and evaluate the action of protein exudates from Mimosa caesalpiniifolia, Leucaena leucocephala, Acacia mangium, and Stylosanthes capitata seeds on the gastrointestinal nematode Haemonchus contortus. The exuded proteins were precipitated, dialyzed, lyophilized, and assessed for their effect on egg hatching and artificial larval exsheathment inhibition. Proteome analysis of the protein extracts was also performed. Although no egg-hatching inhibition was observed, all exudates showed efficacy in inhibiting the larval exsheathment of H. contortus larvae with an EC50 varying from 0.61 to 0.26 mg P mL-1. Proteomic analysis revealed the presence of proteases, protease inhibitors, chitinases, and lectins among other proteins in the exudates. Most of the exuded proteins belong to the oxidative stress/plant defense and energy/carbohydrate metabolism functional clusters. This study concluded that the bioactive proteins from different classes exuded by seeds of M. caesalpiniifolia, L. leucocephala, A. mangium, and S. capitata show stage-specific inhibition against H. contortus.


Subject(s)
Exudates and Transudates/chemistry , Fabaceae/chemistry , Haemonchus/drug effects , Plant Proteins/pharmacology , Seeds/chemistry , Animals , Anthelmintics/chemistry , Anthelmintics/pharmacology , Plant Exudates/chemistry
3.
J Parasitol ; 107(1): 23-28, 2021 01 01.
Article in English | MEDLINE | ID: mdl-33498082

ABSTRACT

Gastrointestinal nematode infection of small ruminants causes losses in livestock production. Plant compounds show promises as alternatives to commercial anthelmintics that have been exerting selective pressures that lead to the development of drug-resistant parasites. Soybean (Glycine max) is an economical value crop, with a higher protein content compared to other legumes. The objective of this study was to evaluate whether the protease inhibitors exuded from the G. max mature seeds have anthelmintic activity against Haemonchus contortus. To obtain the soybean exudates (SEX), mature seeds were immersed in 100 mM sodium acetate buffer, pH 5.0, at 10 C, for 24 hr. Then the naturally released substances present in SEX were collected and exhaustively dialyzed (cutoff 12 kDa) against distilled water. The dialyzed seed exudates (SEXD) were heated at 100 C for 10 min and centrifuged (12,000 g, at 4 C for 15 min). The supernatant obtained was recovered and designated as the heat-treated exudate fraction (SEXDH). The protein content, protease inhibitor activity, and the effect of each fraction on H. contortus egg hatch rate were evaluated. The inhibition extent of SEX, SEXD, and SEXDH on H. contortus egg proteases was 31.1, 42.9, and 63.8%, respectively. Moreover, SEX, SEXD, and SEXDH inhibited the egg hatching with EC50 of 0.175, 0.175, and 0.241 mg ml-1, respectively. Among the commercial protease inhibitors tested, only EDTA and E-64 inhibited the H. contortus hatch rate (79.0 and 28.9%, respectively). We present evidence demonstrating that soybean exudate proteins can effectively inhibit H. contortus egg hatching. This bioactivity is displayed by thermostable proteins and provides evidence that protease inhibitors are a potential candidate for anthelmintic use.


Subject(s)
Exudates and Transudates/chemistry , Glycine max/chemistry , Haemonchus/drug effects , Plant Extracts/pharmacology , Protease Inhibitors/pharmacology , Seeds/chemistry , Animals , Haemonchiasis/parasitology , Haemonchiasis/veterinary , Haemonchus/enzymology , Haemonchus/physiology , Hydrogen-Ion Concentration , Peptide Hydrolases/isolation & purification , Plant Extracts/isolation & purification , Protease Inhibitors/isolation & purification , Sheep , Sheep Diseases/parasitology , Soybean Proteins/chemistry
4.
Expert Rev Proteomics ; 15(12): 967-982, 2018 12.
Article in English | MEDLINE | ID: mdl-30348024

ABSTRACT

INTRODUCTION: Metalloproteinases play key roles in health and disease, by generating novel proteoforms with variable structure and function. Areas covered: This review focuses on the role of endogenous [a Disintegrin and Metalloproteinase (ADAMs), ADAMs with thrombospondin motifs (ADAMTS), and matrix metalloproteinases (MMPs)] and exogenous metalloproteinases in various disease conditions, and describes the application of mass spectrometry-based proteomics to detect qualitative and quantitative changes in protein profiles in tissues and body fluids in disease. Emphasis is placed on the proteomic analysis of exudates collected from affected tissues, including methods that enrich newly generated protein fragments derived from proteolysis in cells, stroma, or extracellular matrix. The use of proteomic analysis of exudates in the study of the local tissue damage induced by metalloproteinases derived from viperid snake venoms is discussed, particularly in relation to extracellular matrix degradation and to the overall pathology of these envenomings. Expert commentary: The information provided by these proteomics approaches is paving the way for the identification of biomarkers based on particular proteolytic signatures associated with different pathologies. Together with other methodological approaches, a comprehensive view of the mechanisms and dynamics of diseases can be achieved. Such basis of knowledge allows for the design of novel diagnostic and therapeutic approaches within the frame of 'precision' or 'personalized' medicine.


Subject(s)
Metalloproteases/analysis , Molecular Diagnostic Techniques/methods , Proteomics/methods , Snake Bites/metabolism , Biomarkers/analysis , Biomarkers/metabolism , Exudates and Transudates/chemistry , Exudates and Transudates/metabolism , Humans , Metalloproteases/metabolism , Snake Bites/pathology
5.
Acta sci. vet. (Online) ; 46: 1-8, 2018. tab
Article in English | VETINDEX | ID: vti-19134

ABSTRACT

Background: Two classifications are used to categorize cavitary effusions using total nucleated cell count (TNCC): protein concentration and pathophysiology of its formation. The aims of the present study were to evaluate the correlation between the TNCC values of cavitary effusions obtained in the automatic and the manual method, and also evaluating the classification methodology.Materials, Methods & Results: Cavitary effusions were analyzed for physical, chemical and cytological aspects, as well as manual and automatic cell counts for the correlation between the traditional methods and those suggested by Stockham & Scott. Bland-Altman regression and Spearman correlation analysis were performed. Of the total, 44 were abdominal effusions (73.3%), 15 thoracic (25%) and 1 pericardial (1.7%). According to the traditional classification, most of the effusions were classified as modified transudates (40%) and according to the classification of Stockham and Scott, as transudates poor in protein (31.7%). The correlation between cell counting techniques between pure, modified and exudate transudates was 0.94, 0.97 and 0.94, respectively, indicating an excellent correlation between the parameters (P = 0.95%).Discussion: Considering the concentration of proteins and CCNT, the effusions classified as modified transudate were mainly caused by neoplastic processes (carcinomas/adenocarcinomas), since there are several mechanisms of their formation, such as large variation of protein concentration. According to the Stockham & Scott classification a unique classification is considered for exfoliative neoplastic effusions, the variation of the protein concentration of the effusion does not alter its classification. In neoplastic effusions, classified as exudates, lymphomas were the most prevalent, and hypercellularity (approximately 150,000 cells / μL) allowed this classification.[...](AU)


Subject(s)
Animals , Cats , Dogs , Exudates and Transudates/chemistry , Cell Count/methods , Flow Cytometry/methods , Horses
6.
Acta sci. vet. (Impr.) ; 46: 1-8, 2018. tab
Article in English | VETINDEX | ID: biblio-1457853

ABSTRACT

Background: Two classifications are used to categorize cavitary effusions using total nucleated cell count (TNCC): protein concentration and pathophysiology of its formation. The aims of the present study were to evaluate the correlation between the TNCC values of cavitary effusions obtained in the automatic and the manual method, and also evaluating the classification methodology.Materials, Methods & Results: Cavitary effusions were analyzed for physical, chemical and cytological aspects, as well as manual and automatic cell counts for the correlation between the traditional methods and those suggested by Stockham & Scott. Bland-Altman regression and Spearman correlation analysis were performed. Of the total, 44 were abdominal effusions (73.3%), 15 thoracic (25%) and 1 pericardial (1.7%). According to the traditional classification, most of the effusions were classified as modified transudates (40%) and according to the classification of Stockham and Scott, as transudates poor in protein (31.7%). The correlation between cell counting techniques between pure, modified and exudate transudates was 0.94, 0.97 and 0.94, respectively, indicating an excellent correlation between the parameters (P = 0.95%).Discussion: Considering the concentration of proteins and CCNT, the effusions classified as modified transudate were mainly caused by neoplastic processes (carcinomas/adenocarcinomas), since there are several mechanisms of their formation, such as large variation of protein concentration. According to the Stockham & Scott classification a unique classification is considered for exfoliative neoplastic effusions, the variation of the protein concentration of the effusion does not alter its classification. In neoplastic effusions, classified as exudates, lymphomas were the most prevalent, and hypercellularity (approximately 150,000 cells / μL) allowed this classification.[...]


Subject(s)
Animals , Cats , Dogs , Flow Cytometry/methods , Cell Count/methods , Exudates and Transudates/chemistry , Horses
7.
J Vet Intern Med ; 30(2): 536-42, 2016.
Article in English | MEDLINE | ID: mdl-26813037

ABSTRACT

BACKGROUND: Pleural effusion is a common cause of dyspnea in cats. N-terminal pro-B-type natriuretic peptide (NT-proBNP) measurement, using a first-generation quantitative ELISA, in plasma and pleural fluid differentiates cardiac from noncardiac causes of pleural effusion. HYPOTHESIS/OBJECTIVES: To determine whether NT-proBNP measurements using second-generation quantitative ELISA and point-of-care (POC) tests in plasma and pleural fluid distinguish cardiac from noncardiac pleural effusions and how results compare to the first-generation ELISA. ANIMALS: Thirty-eight cats (US cohort) and 40 cats (UK cohort) presenting with cardiogenic or noncardiogenic pleural effusion. METHODS: Prospective cohort study. Twenty-one and 17 cats in the US cohort, and 22 and 18 cats in the UK cohort were classified as having cardiac or noncardiac pleural effusion, respectively. NT-proBNP concentrations in paired plasma and pleural fluid samples were measured using second-generation ELISA and POC assays. RESULTS: The second-generation ELISA differentiated cardiac from noncardiac pleural effusion with good diagnostic accuracy (plasma: sensitivity, 95.2%, specificity, 82.4%; pleural fluid: sensitivity, 100%, specificity, 76.5%). NT-proBNP concentrations were greater in pleural fluid (719 pmol/L (134-1500)) than plasma (678 pmol/L (61-1500), P = 0.003), resulting in different cut-off values depending on the sample type. The POC test had good sensitivity (95.2%) and specificity (87.5%) when using plasma samples. In pleural fluid samples, the POC test had good sensitivity (100%) but low specificity (64.7%). Diagnostic accuracy was similar between first- and second-generation ELISA assays. CONCLUSIONS AND CLINICAL IMPORTANCE: Measurement of NT-proBNP using a quantitative ELISA in plasma and pleural fluid or POC test in plasma, but not pleural fluid, distinguishes cardiac from noncardiac causes of pleural effusion in cats.


Subject(s)
Cat Diseases/diagnosis , Heart Diseases/veterinary , Natriuretic Peptide, Brain/metabolism , Peptide Fragments/metabolism , Pleural Effusion/veterinary , Point-of-Care Systems , Animals , Body Fluids/chemistry , Cats , Cohort Studies , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/veterinary , Exudates and Transudates/chemistry , Female , Heart Diseases/complications , Heart Diseases/diagnosis , Male , Natriuretic Peptide, Brain/chemistry , Peptide Fragments/chemistry , Pleural Effusion/diagnosis , Pleural Effusion/etiology , Sensitivity and Specificity
8.
J Endod ; 40(2): 177-81, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24461400

ABSTRACT

INTRODUCTION: This clinical study was conducted to measure the endotoxin levels in infected root canals (RCs) and exudates related to acute apical abscesses (AAAs). In addition, the effectiveness of RC procedures in reducing the endotoxin levels in RCs was monitored. METHODS: Paired samples of infected RCs and exudates from AAAs were collected from 10 subjects by using paper points. RCs samples were collected before (RCS1) and after chemomechanical preparation (CMP) (RCS2), after 17% EDTA (RCS3), and after 30 days of intracanal medication (Ca[OH]2 + chlorhexidine) (RCS4). A turbidimetric kinetic limulus amebocyte lysate assay was used for the measurement of endotoxins. RESULTS: Endotoxins were detected in 100% of the baseline samples of AAAs and RCs (RCS1) with median values of 175 EU/mL and 41.5 EU/mL, respectively (P < .05). After CMP (RCS2), endotoxins were reduced to a median value of 0.54 EU/mL (P < .05). Subsequent irrigation with EDTA (RCS3) failed to present a significant effectiveness in reducing the endotoxin levels (median= 0.37 EU/mL) (P = .07). However, intracanal medication for 30 days (RCS4) reduced endotoxins to median values of 0.03 EU/mL (P < .01). CONCLUSIONS: The present study revealed a strong association between the high levels of endotoxins found in AAAs and RCs collected from the same tooth. Moreover, the effectiveness of CMP in reducing the endotoxin levels from RCs in acute endodontic infection was improved by the use of RC medication.


Subject(s)
Dental Pulp Cavity/microbiology , Dental Pulp Necrosis/microbiology , Endotoxins/analysis , Periapical Abscess/microbiology , Root Canal Preparation/methods , Calcium Hydroxide/therapeutic use , Chlorhexidine/therapeutic use , Dexamethasone/therapeutic use , Drug Combinations , Edetic Acid/therapeutic use , Exudates and Transudates/chemistry , Follow-Up Studies , Formaldehyde/therapeutic use , Gutta-Percha/therapeutic use , Humans , Hydrocortisone/therapeutic use , Limulus Test , Nephelometry and Turbidimetry/methods , Root Canal Filling Materials/therapeutic use , Root Canal Irrigants/therapeutic use , Thymol/analogs & derivatives , Thymol/therapeutic use
9.
West Indian med. j ; West Indian med. j;62(9): 803-807, Dec. 2013. tab
Article in English | LILACS | ID: biblio-1045760

ABSTRACT

OBJECTIVE: Pleural effusion is a common diagnostic and clinical problem. Neoplasms and tuberculosis are the most frequent diagnostic causes of such effusions. Conventional laboratory methods for diagnosis of such effusion are inefficient because tubercle bacilli are rarely seen in direct examinations of pleural fluid. The present study evaluates interleukin-6 (IL-6), gamma interferon (IFN-γ) and adenosine deaminase (ADA) as diagnostic tools in pleural effusion. METHODS: Interleukin-6, IFN-γ and ADA were measured in pleural fluid from the patients, with exudative pleural effusion from tuberculous, malignant and postpneumonic origin and transudative pleural effusion ofsystemic origin in order to evaluate the diagnostic utility ofthese. RESULTS: The three markers were detectable in all effusions with significantly high levels in exudative as compared to transudative effusions. There was a statically significant difference noticed in tuberculous as compared to malignant andpostpneumonic origin and transudative pleural effusion. CONCLUSION: We concluded that IL-6, IFN-γ and ADA levels in pleural effusion are sensitive parameters to differentiate an exudate from a transudate and they can also differentiate exudates of different aetiology. Finally, the results suggest that there is a remarkable difference in production of these three markers in exudative pleural effusions as compared to transudative pleural effusions.


OBJETIVO: El derrame pleural es un problema diagnóstico y clínico común. Las neoplasias y la tuberculosis son las causas más frecuentes en los diagnósticos de tales derrames. Los métodos de laboratorio convencionales para el diagnóstico de tales derrames son ineficientes, porque los bacilos de la tuberculosis raramente se ven en los exámenes directos del líquido pleural. El presente estudio evalúa la interleucina-6, el interferón gamma (IFN-γ) y la adenosina desaminasa (ADA) como herramientas de diagnóstico en el derrame pleural. MÉTODOS: La interleucina-6, el IFN-γ, y la ADA fueron medidos en el líquido pleural de los pacientes con derrame pleural exudativo de origen tuberculoso, maligno y post-pneumónico, y el derrame pleural trasudativo de origen sistémico, con el fin de evaluar la utilidad diagnóstica de éstos. RESULTADOS: Los tres marcadores eran observables en todos los derrames, con niveles significativamente altos en los exudativos en comparación con los trasudativos. Se notó una diferencia estadísticamente significativa en los derrames de origen tuberculoso en comparación con los de origen maligno y postpneumónico, y los derrames pleurales trasudativos. CONCLUSIÓN: Llegamos a la conclusión de que los niveles de IL-6, IFN-Correspondence: Dr M Marie, College of Applied Medical Sciences, Clinical Laboratory Department, King Saud University, PO Box 10219, Riyadh 11433, Kingdom of Saudi Arabia. E-mail: drmmarie.2000@ gmail.com *Equally contributed to the manuscript YADA en el derrame pleural, son parámetros sensibles para diferenciar un derrame pleural exudado de uno trasudado, pudiendo por otra parte ayudar también a distinguir exudados de diferentes etiologías. Finalmente, los resultados sugieren que existe una diferencia notable en la forma en que se producen estos tres marcadores en los derrames efusiones pleurales exudativos en comparación con los derrames pleurales trasudativos.


Subject(s)
Humans , Pleural Effusion/diagnosis , Adenosine Deaminase/analysis , Interleukin-6/analysis , Interferon-gamma/analysis , Exudates and Transudates/chemistry , Biomarkers/analysis
10.
Clinics (Sao Paulo) ; 67(11): 1259-63, 2012 Nov.
Article in English | MEDLINE | ID: mdl-23184200

ABSTRACT

OBJECTIVE: Pleural tuberculosis is the most frequently occurring form of extra pulmonary disease in adults. In up to 40% of cases, the lung parenchyma is concomitantly involved, which can have an epidemiological impact. This study aims to evaluate the pleural and systemic inflammatory response of patients with pleural or pleuropulmonary tuberculosis. METHODS: A prospective study of 39 patients with confirmed pleural tuberculosis. After thoracentesis, a high resolution chest tomography was performed to evaluate the pulmonary involvement. Of the 39 patients, 20 exhibited only pleural effusion, and high resolution chest tomography revealed active associated-pulmonary disease in 19 patients. The total protein, lactic dehydrogenase, adenosine deaminase, vascular endothelial growth factor, interleukin-8, tumor necrosis factor-α, and transforming growth factor-ß(1) levels were quantified in the patient serum and pleural fluid. RESULTS: All of the effusions were exudates with high levels of adenosine deaminase. The levels of vascular endothelial growth factor and transforming growth factor-ß(1) were increased in the blood and pleural fluid of all of the patients with pleural tuberculosis, with no differences between the two forms of tuberculosis. The tumor necrosis factor-α levels were significantly higher in the pleural fluid of the patients with the pleuropulmonary form of tuberculosis. The interleukin-8 levels were high in the pleural fluid of all of the patients, without any differences between the forms of tuberculosis. CONCLUSION: Tumor necrosis factor-α was the single cytokine that significantly increased in the pleural fluid of the patients with pulmonary involvement. However, an overlap in the results does not permit us to suggest that cytokine is a biological marker of concomitant parenchymal involvement. Although high resolution chest tomography can be useful in identifying these patients, the investigation of fast acid bacilli and cultures for M. tuberculosis in the sputum is recommended for all patients who are diagnosed with pleural tuberculosis.


Subject(s)
Biomarkers/analysis , Pleural Effusion/metabolism , Tuberculosis, Pleural/metabolism , Adenosine Deaminase/analysis , Adult , Cytokines/analysis , Disease Progression , Enzyme-Linked Immunosorbent Assay , Exudates and Transudates/chemistry , Humans , Middle Aged , Oxidoreductases/analysis , Pleural Effusion/diagnostic imaging , Prospective Studies , Radiography , Transforming Growth Factor beta1/analysis , Tuberculosis, Pleural/diagnostic imaging , Tuberculosis, Pulmonary/metabolism , Tumor Necrosis Factor-alpha/analysis , Vascular Endothelial Growth Factor A/analysis , Young Adult
11.
Clinics ; Clinics;67(11): 1259-1263, Nov. 2012. ilus, tab
Article in English | LILACS | ID: lil-656714

ABSTRACT

OBJECTIVE: Pleural tuberculosis is the most frequently occurring form of extra pulmonary disease in adults. In up to 40% of cases, the lung parenchyma is concomitantly involved, which can have an epidemiological impact. This study aims to evaluate the pleural and systemic inflammatory response of patients with pleural or pleuropulmonary tuberculosis. METHODS: A prospective study of 39 patients with confirmed pleural tuberculosis. After thoracentesis, a high resolution chest tomography was performed to evaluate the pulmonary involvement. Of the 39 patients, 20 exhibited only pleural effusion, and high resolution chest tomography revealed active associated-pulmonary disease in 19 patients. The total protein, lactic dehydrogenase, adenosine deaminase, vascular endothelial growth factor, interleukin-8, tumor necrosis factor-α, and transforming growth factor-β1 levels were quantified in the patient serum and pleural fluid. RESULTS: All of the effusions were exudates with high levels of adenosine deaminase. The levels of vascular endothelial growth factor and transforming growth factor-β1 were increased in the blood and pleural fluid of all of the patients with pleural tuberculosis, with no differences between the two forms of tuberculosis. The tumor necrosis factor-α levels were significantly higher in the pleural fluid of the patients with the pleuropulmonary form of tuberculosis. The interleukin-8 levels were high in the pleural fluid of all of the patients, without any differences between the forms of tuberculosis. CONCLUSION: Tumor necrosis factor-α was the single cytokine that significantly increased in the pleural fluid of the patients with pulmonary involvement. However, an overlap in the results does not permit us to suggest that cytokine is a biological marker of concomitant parenchymal involvement. Although high resolution chest tomography can be useful in identifying these patients, the investigation of fast acid bacilli and cultures for M. tuberculosis in the sputum is recommended for all patients who are diagnosed with pleural tuberculosis.


Subject(s)
Adult , Humans , Middle Aged , Young Adult , Biomarkers/analysis , Pleural Effusion/metabolism , Tuberculosis, Pleural/metabolism , Adenosine Deaminase/analysis , Cytokines/analysis , Disease Progression , Enzyme-Linked Immunosorbent Assay , Exudates and Transudates/chemistry , Oxidoreductases/analysis , Prospective Studies , Pleural Effusion , Transforming Growth Factor beta1/analysis , Tuberculosis, Pleural , Tuberculosis, Pulmonary/metabolism , Tumor Necrosis Factor-alpha/analysis , Vascular Endothelial Growth Factor A/analysis
12.
J Endod ; 37(5): 602-7, 2011 May.
Article in English | MEDLINE | ID: mdl-21496656

ABSTRACT

INTRODUCTION: Root canal contents are potent stimuli for proinflammatory cytokines involved in apical periodontitis. This study investigated target gram-negative bacterial species and endotoxins in primary endodontic infection with apical periodontitis, determined their antigenicity against macrophages through the levels of PGE(2), and evaluated their relationship with clinical findings. METHODS: Samples were taken from 21 root canals with primary infection and apical periodontitis by using paper points. Polymerase chain reaction (16S rDNA) was used for bacterial detection and limulus amebocyte lysate assay for endotoxin measurement. Levels of prostaglandin E2 (PGE(2)) were measured by enzyme-linked immunosorbent assay (Duoset Kit; R&D, Minneapolis, MN). RESULTS: Prevotella nigrescens (13/21), Fusobacterium nucleatum (6/21), and Porphyromonas endodontalis (6/21) were the most frequently observed species. A positive association was found between F. nucleatum and P. endodontalis (P < .05). A correlation was found between the number of gram-negative bacterial species and the levels of endotoxins, such as PGE(2) (P < .05). Higher levels of endotoxin were detected in teeth with exudation, whereas elevated levels of PGE(2) were found in teeth with tenderness to percussion and pain on palpation. CONCLUSIONS: Our findings imply an additive effect between the number of gram-negative bacterial species involved in endodontic infection regarding the induction of proinflammatory cytokine by macrophage cells. Moreover, teeth with clinical symptomatology were related to higher levels of endotoxins and PGE(2) secretion.


Subject(s)
Antigens, Bacterial/immunology , Dental Pulp Necrosis/microbiology , Dinoprostone/immunology , Gram-Negative Bacterial Infections/immunology , Macrophages/immunology , Adolescent , Adult , Aged , Cell Culture Techniques , Cell Survival/physiology , Cytokines/analysis , Cytokines/immunology , Dental Pulp Necrosis/immunology , Dinoprostone/analysis , Endotoxins/analysis , Endotoxins/immunology , Exudates and Transudates/chemistry , Fusobacterium Infections/immunology , Fusobacterium nucleatum/immunology , Humans , Inflammation Mediators/immunology , Middle Aged , Periapical Periodontitis/immunology , Periapical Periodontitis/microbiology , Porphyromonas endodontalis/immunology , Porphyromonas gingivalis/immunology , Prevotella intermedia/immunology , Prevotella nigrescens/immunology , RNA, Bacterial/analysis , Young Adult
13.
J Proteome Res ; 10(4): 1987-2005, 2011 Apr 01.
Article in English | MEDLINE | ID: mdl-21306181

ABSTRACT

Tissue damage analysis by traditional laboratory techniques is problematic. Proteomic analysis of exudates collected from affected tissue constitutes a powerful approach to assess tissue alterations, since biomarkers associated with pathologies can be identified in very low concentrations. In this study we proteomically explore the pathological effects induced by the venom of the viperid snake Bothrops asper in the gastrocnemius muscle of mice. Predominant proteins identified in the exudates included intracellular proteins, plasma proteins, extracellular matrix proteins and cell membrane-associated proteins. The presence of such proteins indicates cytotoxicity, plasma exudation, extracellular matrix degradation and shedding of membrane proteins. Some of these proteins may represent useful biomarkers for myonecrosis and microvascular damage. The effect of fucoidan, an inhibitor of myotoxic phospholipases A(2), and batimastat, an inhibitor of metalloproteinases, on the pathological effects induced by B. asper venom were also investigated. Fucoidan reduced the presence of intracellular proteins in exudates, whereas batimastat reduced the amount of relevant extracellular matrix proteins. The combination of these inhibitors resulted in the abrogation of the most relevant pathological effects of this venom. Thus, proteomic analysis of exudates represents a valuable approach to assess the characteristics of tissue damage in pathological models and the success of therapeutic interventions.


Subject(s)
Biomarkers/analysis , Exudates and Transudates/chemistry , Muscle, Skeletal/drug effects , Muscle, Skeletal/pathology , Proteome/analysis , Snake Bites/pathology , Snake Venoms/toxicity , Animals , Anticoagulants/chemistry , Anticoagulants/metabolism , Bothrops , Female , Male , Mass Spectrometry/methods , Metalloendopeptidases/antagonists & inhibitors , Metalloendopeptidases/metabolism , Mice , Muscle, Skeletal/chemistry , Necrosis/pathology , Phenylalanine/analogs & derivatives , Phenylalanine/chemistry , Phenylalanine/metabolism , Polysaccharides/chemistry , Polysaccharides/metabolism , Protease Inhibitors/chemistry , Protease Inhibitors/metabolism , Proteomics/methods , Snake Venoms/chemistry , Thiophenes/chemistry , Thiophenes/metabolism
14.
Int J Clin Exp Pathol ; 4(1): 74-84, 2010 Dec 20.
Article in English | MEDLINE | ID: mdl-21228929

ABSTRACT

The effect of exogenous Gal-1 on cellular response and adhesion molecule expression was investigated in a classical model of acute inflammation induced by zymosan. C57BL6 mice, treated or not with human recombinant (hr) Gal-1, received i.p. injection of zymosan and peritoneal exudate, blood and mesentery were processed for cellular, biochemical, light and electron microscopic analysis after 4 and 24 h. Zymosan peritonitis provoked the expected signs of inflammation at 4 h, including a significant increase in extravasated PMNs in the mesentery and peritoneal exudate, mirrored by blood neutrophilia. These changes subsided after 24 h. Ultrastructural immunocytochemical analysis of PMNs showed significant Gal-1 expression and co-localization with L-selectin and ß2-integrin in the plasma membrane and cytoplasm. Pharmacological treatment with hrGal-1 at 4 h produced an inhibition of PMN migration, associated with diminished expression of adhesion molecules, particularly ß2-integrin, and TNF-α and IL-1ß release by peritoneal cells. At 24 h, Gal-1 induced an increase in mononuclear phagocytic cell recruitment. In conclusion, our data propose an important mechanism of anti-inflammatory action of Gal-1, initially by modulation of pro-inflammatory cytokine release and PMN migration through an imbalance between adhesion molecule expression and, later, by promoting monocyte-macrophage recruitment.


Subject(s)
Cell Adhesion Molecules/metabolism , Cytokines/metabolism , Galectin 1/pharmacology , Leukocytes/drug effects , Animals , CD18 Antigens/metabolism , Cell Migration Assays, Leukocyte , Cell Movement/drug effects , Disease Models, Animal , Exudates and Transudates/chemistry , Exudates and Transudates/metabolism , Galectin 1/metabolism , Humans , Intercellular Adhesion Molecule-1 , L-Selectin/metabolism , Leukocytes/metabolism , Leukocytes/pathology , Male , Mesentery/drug effects , Mesentery/metabolism , Mesentery/pathology , Mice , Mice, Inbred C57BL , Monocytes/drug effects , Monocytes/metabolism , Monocytes/pathology , Neutrophils/drug effects , Neutrophils/metabolism , Neutrophils/pathology , Peritonitis/immunology , Peritonitis/metabolism , Peritonitis/pathology , Recombinant Proteins
15.
J Proteome Res ; 8(11): 5120-31, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19764775

ABSTRACT

In light of the complexity of wound tissue, proteomic analysis may not clearly reveal the nature of the wound or the processes involved in healing. However, exudate associated with wounds may provide a "window" on cellular events leading to the development of the wound and/or its healing. In this investigation we performed proteomic analysis on wound exudates from muscular wounds in mice caused by two very different types of snake venom toxins: BaP1, a snake venom metalloproteinase and Mtx-I, a snake venom phospholipase A2. Proteomic analysis of the exudates associated with these wounds clearly differentiated them and offered new perspectives on functional mechanisms by which these toxins cause tissue damage. In the case of wounds caused by the metalloproteinase, there was evidence of degradation of nonfibrillar collagens whereas the phospholipase wound exudate was noted by the presence of fibrillar collagen type I, apolipoproteins A-I, A-IV, and E, and fibronectin. These results suggest that the hemorrhage caused by snake venom metalloproteinases may be associated with the degradation of specific extracellular matrix proteins which play a role in matrix/capillary stabilization and that release of apolipoproteins from their complexes may be involved with the dysfunctional hemostasis observed following snake envenoming.


Subject(s)
Exudates and Transudates/chemistry , Metalloendopeptidases , Muscle, Skeletal , Phospholipases A2 , Proteomics/methods , Snake Venoms , Wound Healing/physiology , Animals , Bothrops , Chromatography, Liquid/methods , Extracellular Matrix Proteins/metabolism , Keratins/metabolism , Mass Spectrometry/methods , Metalloendopeptidases/pharmacology , Metalloendopeptidases/toxicity , Mice , Molecular Sequence Data , Muscle, Skeletal/drug effects , Muscle, Skeletal/metabolism , Muscle, Skeletal/pathology , Phospholipases A2/pharmacology , Phospholipases A2/toxicity , Skin/cytology , Skin/drug effects , Skin/metabolism , Skin/pathology , Snake Bites , Snake Venoms/pharmacology , Snake Venoms/toxicity , Tandem Mass Spectrometry/methods
16.
Clinics (Sao Paulo) ; 64(9): 891-5, 2009.
Article in English | MEDLINE | ID: mdl-19759883

ABSTRACT

OBJECTIVE: The aim of this study was to identify the participation of the coagulation system in the differential diagnosis of pleural effusions. INTRODUCTION: Imbalance between immunologic and metabolic factors triggers a sequence of events resulting in pleural reactions and accumulation of fluid. The coagulation system, which is fundamental for the maintenance of homeostasis, contributes to the inflammatory process responsible for pleural effusions, and participates in cellular proliferation and migration as well as in the synthesis of inflammatory mediators. METHODS: We evaluated the laboratory profile of coagulation and fibrinolysis in 54 pleural fluids (15 transudates and 39 exudates). RESULTS: The coagulation system acts according to the pathophysiologic mechanisms involved in the development of pleural effusions. In inflammatory effusions (exudates), there is activation of coagulation with increased levels of fragment 1+2 and thrombin-antithrombin complex in addition to reduction of fibrinogen levels due to fibrinolysis and fibrin tissue incorporation. As a consequence, there is activation of the fibrinolytic system with increased levels of fibrin degradation products, including the D-dimer. These changes are not sufficient for differentiation of different subgroups of exudates. In transudates, these events were observed to a lesser degree. CONCLUSION: The coagulation system plays an important role in the development of pleural diseases. Coagulation tests show differences between transudates and exudates but not among exudate subgroups. Understanding the physiopathological mechanisms of pleural disorders may help to define new diagnostic and therapeutic approaches.


Subject(s)
Blood Coagulation/physiology , Exudates and Transudates/chemistry , Fibrinolysin/analysis , Pleural Effusion/diagnosis , Diagnosis, Differential , Humans , Pleural Effusion/blood , Pleural Effusion/etiology
17.
Clinics ; Clinics;64(9): 891-895, 2009. graf
Article in English | LILACS | ID: lil-526329

ABSTRACT

OBJECTIVE: The aim of this study was to identify the participation of the coagulation system in the differential diagnosis of pleural effusions. INTRODUCTION: Imbalance between immunologic and metabolic factors triggers a sequence of events resulting in pleural reactions and accumulation of fluid. The coagulation system, which is fundamental for the maintenance of homeostasis, contributes to the inflammatory process responsible for pleural effusions, and participates in cellular proliferation and migration as well as in the synthesis of inflammatory mediators. METHODS: We evaluated the laboratory profile of coagulation and fibrinolysis in 54 pleural fluids (15 transudates and 39 exudates). RESULTS: The coagulation system acts according to the pathophysiologic mechanisms involved in the development of pleural effusions. In inflammatory effusions (exudates), there is activation of coagulation with increased levels of fragment 1+2 and thrombin-antithrombin complex in addition to reduction of fibrinogen levels due to fibrinolysis and fibrin tissue incorporation. As a consequence, there is activation of the fibrinolytic system with increased levels of fibrin degradation products, including the D-dimer. These changes are not sufficient for differentiation of different subgroups of exudates. In transudates, these events were observed to a lesser degree. CONCLUSION: The coagulation system plays an important role in the development of pleural diseases. Coagulation tests show differences between transudates and exudates but not among exudate subgroups. Understanding the physiopathological mechanisms of pleural disorders may help to define new diagnostic and therapeutic approaches.


Subject(s)
Humans , Blood Coagulation/physiology , Exudates and Transudates/chemistry , Fibrinolysin/analysis , Pleural Effusion/diagnosis , Diagnosis, Differential , Pleural Effusion/blood , Pleural Effusion/etiology
18.
Ginecol Obstet Mex ; 76(1): 3-8, 2008 Jan.
Article in Spanish | MEDLINE | ID: mdl-18798390

ABSTRACT

OBJECTIVE: To evaluate the secretion of interleukin 6 (IL-6) in cervicovaginal fluid in a pseudocohort that emulates the evolution of the labor. MATERIAL AND METHOD: Samples of cervicovaginal fluid of patients with 20 to 40 weeks of gestation were taken, patients were classified in 5 strata of the development of the labor. Each stratum reflects the progressive activation of the uterine activity and the cervical changes. To each sample was made determination of IL-6 by means of the system Multiplex. The samples of patient with infection data were eliminated. RESULTS: 173 samples were included distributed in each one of the five strata. Basal secretion exists of IL-6 to the cervicovaginal fluid along the gestation, that doesn't modify until the active labor appears. Only when the uterine activity is manifested in form to regulate and effective, very significant increase is documented in the concentration of the IL-6 in the cervicovaginal fluid. CONCLUSIONS: The IL-6 it is a proinflammatory cytokine that increases in a specific way in the moment in that the effective uterine activity begins, thus, it is an excellent candidate to be evaluated as marker presage of the event of normal labor and preterm.


Subject(s)
Exudates and Transudates/chemistry , Interleukin-6/analysis , Interleukin-6/metabolism , Labor, Obstetric/metabolism , Cervix Uteri , Female , Humans , Pregnancy , Vagina
19.
Aesthetic Plast Surg ; 31(5): 514-8, 2007.
Article in English | MEDLINE | ID: mdl-17659406

ABSTRACT

BACKGROUND: This study aimed to analyze the composition of postabdominoplasty seroma fluid at different intervals, compare it with blood and lymph, and determine whether it meets the criteria for being considered an exudate. METHODS: The study enrolled 18 female patients with postabdominoplasty seroma diagnosed by clinical and ultrasound evaluation. All the patients had a Matarasso type 4 anterior abdominal wall deformity. None of the patients were overweight, and none had comorbidities. They all underwent a classical abdominoplasty procedure. Fluid samples were taken from the drains between postoperative days 5 and 7, and from needle aspiration between postoperative days 15 and 20. The fluids were assayed in the clinical laboratory at the University of Chile Clinical Hospital for chemical and cellular composition. Blood, lymph, and seroma values were compared by independent group analysis using a Tukey multiple comparison test with an alpha error of 0.05. RESULTS: The total protein, lactate dehydrogenase (LDH), and cholesterol levels for the early and late seroma fluids were lower than in the blood, but higher than in the lymph. The total protein seroma-to-plasma ratio was approximately 0.5; the LDH seroma-to-plasma ratio was approximately 0.6; and the cholesterol seroma-to-plasma ratio was 0.32. The platelet level was very low in the late seroma fluid, showing no statistical differences with the lymph level. The leukocyte level was low in the seroma fluid, with a higher percentage of neutrophils than found in the blood or the lymph. CONCLUSIONS: The serous fluid formed under the flap after an abdominoplasty seems to be an exudate. In the early postoperative period, it is an inflammatory exudate that slowly turns into an exudate with some characteristics similar to those of lymph.


Subject(s)
Exudates and Transudates/chemistry , Lipectomy/adverse effects , Seroma/etiology , Abdominal Wall/surgery , Drainage/methods , Female , Humans , Postoperative Complications/etiology
20.
J Dent Res ; 85(9): 834-8, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16931867

ABSTRACT

Calcium (Ca), inorganic phosphorus (P(i)), and fluoride (F) concentrations are low in the whole plaque biofilm formed under exposure to sucrose. It was hypothesized that this would be reflected in the biofilm fluid, where these low values should greatly influence the de/remineralization process. Dental biofilms were formed in situ over enamel blocks mounted in palatal appliances and exposed 8 times/day to distilled water, glucose+fructose, or sucrose solutions for 14 days. While Ca, P(i), and F concentrations in the whole biofilms were significantly lower in the glucose+fructose and sucrose groups, no effect on biofilm fluid was observed, even after a cariogenic challenge. An increase in whole biofilm mineral ions was observed 24 hrs after the carbohydrate treatments were suspended, but this effect was also not observed in the fluid. These results suggest that there is a homeostatic mechanism that maintains biofilm fluid mineral ion concentration, regardless of its total concentration in the whole biofilm.


Subject(s)
Biofilms , Cariogenic Agents/adverse effects , Dental Plaque/chemistry , Tooth Demineralization/metabolism , Tooth Remineralization , Adult , Analysis of Variance , Calcium/analysis , Cariostatic Agents/therapeutic use , Cross-Over Studies , Dental Plaque/metabolism , Double-Blind Method , Exudates and Transudates/chemistry , Fluorides/analysis , Fructose/adverse effects , Glucose/adverse effects , Homeostasis , Humans , Hydrogen-Ion Concentration , Ion-Selective Electrodes , Phosphorus/analysis , Sodium Fluoride/therapeutic use , Statistics, Nonparametric , Sucrose/adverse effects , Tooth Demineralization/chemically induced
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