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1.
Int Endod J ; 51(8): 901-911, 2018 Aug.
Article in English | MEDLINE | ID: mdl-29397005

ABSTRACT

AIM: To evaluate the antimicrobial action of an irrigant containing silver nanoparticles in an aqueous vehicle (AgNp), sodium hypochlorite and chlorhexidine against Enterococcus faecalis biofilm and infected dentinal tubules. METHODOLOGY: Bovine dentine blocks were used for E. faecalis biofilm development for 21 days and irrigated with 94 ppm AgNp solution, 2.5% NaOCl and 2% chlorhexidine for 5, 15 and 30 min. For infection of dentinal tubules with E. faecalis, dentine specimens from bovine incisors were submitted to a contamination protocol over 5 days, with eight centrifugation cycles on every alternate day, and irrigated with the same solutions and time intervals used for the biofilm. The specimens were stained with the Live/Dead technique and evaluated using a confocal laser scanning microscope (CLSM). The bioImage_L software was used for measurement of the total biovolume of biofilm in µm3 and percentage of viable bacteria (green cells) in biofilm and in dentinal tubules found after the irrigation. Statistical analyses were performed using Kruskal-Wallis and Dunn's tests for quantification of viable cells in biofilm, the Friedman test for comparisons of viable bacteria in dentinal tubules in different areas of the root canal and the Mann-Whitney U-test to compare the action of the irrigants between the two methods (P < 0.05). RESULTS: The AgNp solution eliminated fewer bacteria, but was able to dissolve more biofilm compared with chlorhexidine (P < 0.05). NaOCl had the greatest antimicrobial activity and biofilm dissolution capacity. AgNp solution had less antimicrobial action in infected dentinal tubules compared with NaOCl (P < 0.05). The AgNp solution after 5 min was more effective in eliminating planktonic bacteria in dentinal tubules than in biofilm, but at 30 min fewer viable bacteria were observed in the biofilm compared with intratubular dentine (P < 0.05). CONCLUSIONS: AgNp irrigant was not as effective against E. faecalis compared to solutions commonly used in root canal treatment. NaOCl is appropriate as an irrigant because it was effective in disrupting biofilm and in eliminating bacteria in biofilms and in dentinal tubules.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Biofilms/drug effects , Enterococcus faecalis/drug effects , Metal Nanoparticles/therapeutic use , Root Canal Irrigants/pharmacology , Silver/therapeutic use , Animals , Anti-Bacterial Agents/administration & dosage , Cattle , Gram-Positive Bacterial Infections/drug therapy , Metal Nanoparticles/administration & dosage , Root Canal Irrigants/therapeutic use , Silver/administration & dosage , Silver/pharmacology
2.
J Viral Hepat ; 23(2): 116-22, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26400151

ABSTRACT

Hepatitis C virus (HCV) is the main cause of chronic liver disease, cirrhosis and hepatocellular carcinoma (HCC) worldwide. The risk to develop HCC increases with the severity of liver inflammation and fibrosis. Long pentraxin 3 (PTX3) is a soluble pattern-recognition receptor produced by phagocytes and nonimmune cells at sites of inflammation or injury. The aim of the present study was to determine the association of PTX3 polymorphisms and its plasma levels with HCC occurrence among patients with HCV. Samples from 524 patients with chronic hepatitis C were evaluated in this study. Two polymorphisms (rs1840680 and rs2305619) in the PTX3 gene were determined by real-time PCR. PTX3 plasma levels were measured by Enzyme-linked Immunosorbent Assay (ELISA). Our data show a significant association between PTX3 polymorphisms and HCC occurrence in univariate and multivariate analysis (P = 0.024). Patients with HCC had higher PTX3 plasma levels compared to individuals with mild or severe fibrosis (P < 0.0001 and P = 0.002, respectively). In addition, PTX3 rs2305619 polymorphism and plasma levels were correlated with Child-Pugh scores B and C in HCC individuals. PTX3 seems to be a risk factor for HCC occurrence in chronic hepatitis C. This is the first study that evaluates PTX3 in the context of hepatitis C.


Subject(s)
C-Reactive Protein/genetics , C-Reactive Protein/metabolism , Carcinoma, Hepatocellular/blood , Hepatitis C, Chronic/virology , Liver Neoplasms/blood , Serum Amyloid P-Component/genetics , Serum Amyloid P-Component/metabolism , Carcinoma, Hepatocellular/virology , Enzyme-Linked Immunosorbent Assay , Female , Hepacivirus , Humans , Liver Cirrhosis/virology , Liver Neoplasms/virology , Male , Middle Aged , Polymorphism, Single Nucleotide/genetics , Real-Time Polymerase Chain Reaction , Risk Factors
3.
Cytokine ; 62(3): 421-5, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23602201

ABSTRACT

Hepatitis C virus (HCV) is the main cause of chronic liver disease, cirrhosis and hepatocellular carcinoma (HCC) worldwide. The risk for the development of HCC increases with the severity of liver inflammation and fibrosis. The hepatic inflammation caused by HCV involves host regulatory immune response, which is mediated by cytokines with anti-viral role upon the interaction of viral polypeptides with innate and adaptive immunity. Two cytokines; tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) play key roles in the regulation of cellular immune response in HCV infection. The aim of the present study was to determine the levels of IL-10 and TNF-α, as well as the ratio of TNF-α and IL-10 serum levels in patients with HCV and HCC caused by HCV (HCC-HCV). The study included 173 patients with chronic HCV. TNF-α and IL-10 serum levels were measured by ELISA (R&D Systems, Inc.). In the present study, 54 patients presented liver mild fibrosis, 68 had severe fibrosis and 51 patients had HCC. After adjustment in the multivariate regression analysis, the following variables remained significantly associated with HCC-HCV occurrence: diabetes (p=0.012 OR 10.44 CI 1.66-65.60), IL-10 lower levels (p<0.0001 OR 0.83 CI 0.78-0.89) and TNF-α higher levels (p<0.0001 OR 1.19 CI 1.11-1.28). Individuals with HCC presented higher TNF-α/IL-10 ratio than those with fibrosis grade F4, F3 or F0+F1+F2 (p=0.0003, p<0.0001, p<0.0001, respectively). Patients with HCC were associated to higher index TNF-α/IL-10 ratio, suggesting that the unbalanced production of these cytokines may represent progression to the liver disease severity in HCV infected patients.


Subject(s)
Carcinoma, Hepatocellular/blood , Carcinoma, Hepatocellular/complications , Hepatitis C, Chronic/blood , Hepatitis C, Chronic/complications , Interleukin-10/blood , Liver Neoplasms/blood , Tumor Necrosis Factor-alpha/blood , Carcinoma, Hepatocellular/genetics , Female , Hepatitis C, Chronic/genetics , Humans , Liver Cirrhosis/blood , Liver Cirrhosis/genetics , Liver Neoplasms/complications , Liver Neoplasms/genetics , Male , Middle Aged , Multivariate Analysis , Risk Factors
4.
Blood Cells Mol Dis ; 44(4): 224-8, 2010 Apr 15.
Article in English | MEDLINE | ID: mdl-20172753

ABSTRACT

Vasoocclusive crisis (VOC) is the major cause of morbidity and mortality in sickle cell anemia (SCA), which is caused by the occlusion of blood vessels, followed by ischemia or infarct, resulting in progressive damage to organs. However, this clinical manifestation is variable, indicating that this process could be influenced by modifier genes. The gene MBL2 which codes for mannose-binding lectin (MBL) has been associated with modifications in the progression of infectious and inflammatory vascular diseases. The aim of this study was to determine the frequency of the polymorphisms of exon 1 (alleles A/O) and promoter region -221 (alleles Y/X) of MBL2 in children with SCA and to verify their association with VOC. The determination of the polymorphism of exon 1 and the promoter region of MBL2 was performed by SYBR GREEN((R)) and Taqman((R)) system, respectively. In the patients with SCA, the frequency of the genotype related to high production of MBL was 0.46 (YA/YA) and for intermediate/low production was 0.54 (YA/XA, XA/XA, YA/YO, XA/YO, YO/YO). The frequency of the genotypes and haplotypes of MBL2 in patients with SCA did not differ from control individuals. The populations were in Hardy-Weinberg equilibrium. The patients were divided into two groups. The groups were separated by the frequency of VOC, which was defined by the total of VOC episodes divided by the age of the children at the end of this study. Since, we choose a cut point in FVOC <1 (n=48) (which we considered of mild presentation of disease) and FVOC >or=1 (n=39) (higher severity). In children with SCA, the frequency of the genotypes of MBL2 of intermediate/low expression for MBL was associated with FVOC >or=1 (p=0.0188 OR=3.15 CI=1.19-8.50). The results suggest that MBL2 polymorphism at promoter and first exon of MBL2 associated with low serum levels and structural alterations of MBL could modify the phenotype of the child with SCA related to VOC.


Subject(s)
Anemia, Sickle Cell/complications , Mannose-Binding Lectin/genetics , Vascular Diseases/etiology , Alleles , Anemia, Sickle Cell/genetics , Child , Child, Preschool , Exons/genetics , Female , Genetic Predisposition to Disease , Genotype , Haplotypes/genetics , Humans , Infant , Infarction/etiology , Infarction/genetics , Ischemia/etiology , Ischemia/genetics , Male , Mannose-Binding Lectin/physiology , Promoter Regions, Genetic/genetics , Severity of Illness Index , Vascular Diseases/genetics
5.
Int J Immunogenet ; 36(6): 377-81, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19703233

ABSTRACT

We studied the association of the mannose-binding lectin-2 (MBL2) polymorphism with anti-thyroid antibodies (ATA) in hepatitis C virus (HCV)-infected Brazilian patients (n = 162) and 124 healthy volunteers screened for ATA. Our results showed that patients with ATA had higher frequency of genotype 00 than controls. MBL may play a role as disease modifier in HCV infection.


Subject(s)
Autoimmunity/genetics , Hepacivirus/immunology , Hepatitis C/blood , Mannose-Binding Lectin/genetics , Polymorphism, Genetic , Thyroid Gland/immunology , Adolescent , Adult , Aged , Brazil , Case-Control Studies , Female , Hepatitis C/genetics , Humans , Male , Middle Aged
7.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;42(4): 353-357, Apr. 2009. graf, tab
Article in English | LILACS | ID: lil-509168

ABSTRACT

The clinical heterogeneity observed in leptospirosis may be associated with host factors or bacteria virulence. Human serum mannose-binding lectin (MBL) recognizes many pathogens, and low levels of this lectin are associated with susceptibility to infection. MBL is also implicated in the modulation of the inflammatory process. We determined the levels of serum MBL during leptospirosis infection. A double-antibody sandwich ELISA was used to detect the immunoreactive serum MBL. The ELISA plates were coated with monoclonal antibody to MBL and bound MBL or recombinant human MBL were detected by rabbit anti-human MBL serum. HRPO-conjugated goat anti-rabbit antibody was used for detection of the reaction. Two groups of patients seen at referral hospitals in Recife, PE, Brazil, were divided according to the year of infection, 2001 (N = 61) or 2002 (N = 57) and compared in terms of disease severity and levels of serum MBL. A group of healthy volunteers (N = 97) matched by age, gender, and ethnic background was used as control. Patients infected in 2001 had more severe outcomes than those infected in 2002, including jaundice, hemorrhage, respiratory alteration, and renal complication (P = 0.0009; chi-square test). The frequency of patients producing serum MBL >1000 ng/mL was higher in the 2001 group than in the 2002 and control groups (P < 0.01), suggesting an association of MBL level with disease severity. The involvement of MBL and genetic variation of the MBL2 gene should be further evaluated to establish the role of this lectin in the pathogenesis of leptospirosis.


Subject(s)
Adolescent , Animals , Female , Humans , Male , Rabbits , Young Adult , Leptospirosis/blood , Mannose-Binding Lectin/blood , Antibodies, Monoclonal/blood , Case-Control Studies , Enzyme-Linked Immunosorbent Assay/methods , Leptospirosis/complications , Leptospirosis/immunology , Mannose-Binding Lectin/immunology , Severity of Illness Index , Young Adult
8.
Braz J Med Biol Res ; 42(4): 353-7, 2009 Apr.
Article in English | MEDLINE | ID: mdl-19330263

ABSTRACT

The clinical heterogeneity observed in leptospirosis may be associated with host factors or bacteria virulence. Human serum mannose-binding lectin (MBL) recognizes many pathogens, and low levels of this lectin are associated with susceptibility to infection. MBL is also implicated in the modulation of the inflammatory process. We determined the levels of serum MBL during leptospirosis infection. A double-antibody sandwich ELISA was used to detect the immunoreactive serum MBL. The ELISA plates were coated with monoclonal antibody to MBL and bound MBL or recombinant human MBL were detected by rabbit anti-human MBL serum. HRPO-conjugated goat anti-rabbit antibody was used for detection of the reaction. Two groups of patients seen at referral hospitals in Recife, PE, Brazil, were divided according to the year of infection, 2001 (N = 61) or 2002 (N = 57) and compared in terms of disease severity and levels of serum MBL. A group of healthy volunteers (N = 97) matched by age, gender, and ethnic background was used as control. Patients infected in 2001 had more severe outcomes than those infected in 2002, including jaundice, hemorrhage, respiratory alteration, and renal complication (P = 0.0009; chi-square test). The frequency of patients producing serum MBL >1000 ng/mL was higher in the 2001 group than in the 2002 and control groups (P < 0.01), suggesting an association of MBL level with disease severity. The involvement of MBL and genetic variation of the MBL2 gene should be further evaluated to establish the role of this lectin in the pathogenesis of leptospirosis.


Subject(s)
Leptospirosis/blood , Mannose-Binding Lectin/blood , Adolescent , Animals , Antibodies, Monoclonal/blood , Case-Control Studies , Enzyme-Linked Immunosorbent Assay/methods , Female , Humans , Leptospirosis/complications , Leptospirosis/immunology , Male , Mannose-Binding Lectin/immunology , Rabbits , Severity of Illness Index , Young Adult
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