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1.
Iran J Basic Med Sci ; 22(1): 58-64, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30944709

RESUMO

Objectives: Pseudomonas aeruginosa infections such as keratitis are considered among the major health problems worldwide due to the complexity of pathogenesis and antibiotic resistance crisis, thus, finding new effective approaches for prevention and treatment of the infections seem to be still vital. In this report, we aimed to investigate the therapeutic effects of topical administration of the antibodies against type a and b-flagellin (FLA and FLB) in Pseudomonas keratitis model of infection in mice. Materials and Methods: Scratched corneas of mice were treated with approximately 107 CFUs/eye of PAK and/or PAO1 strains of P. aeruginosa. Specific IgG to FLA, FLB or divalent flagellin were topically applied to the infected corneas for 20 min, 24, and 36 hr post-infection. The bacterial burden and myeloperoxidase activity (as a marker for polymorphonuclears (PMNs) infiltration) were determined in the corneas. The biological activity of the anti-FLA and FLB IgG was evaluated in vitro by opsonophagocytosis test. Results: Compared to other treated corneas, divalent anti-flagellin IgG treatment showed a significant decrease in the bacterial CFUs and myeloperoxidase activity in the infected corneas (P<0.05). Results of opsonophagocytosis revealed that the specific antibodies raised against FLA and FLB had more potent opsonic killing activity on their homologous strains as compared with control group (P<0.05). Conclusion: It appears that in P. aeruginosa keratitis, topical administration of the combined antibodies likely via decreasing the bacterial load, and PMNs infiltration as well as increasing opsonophagocytosis could lead to dramatic improvement of the infected corneas.

2.
BMC Res Notes ; 12(1): 28, 2019 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-30646938

RESUMO

OBJECTIVE: The present study aimed to determine the prevalence of virulence factors and antimicrobial resistance profile of Pseudomonas aeruginosa strains isolated from Iranian burn patients. RESULTS: This cross-sectional study performed on 100 P. aeruginosa isolates which were recovered from burn wound specimens in 2014-2015. All presumptive isolates were identified by standard microbiologic tests. Antimicrobial susceptibility test was carried out by disk diffusion method. The presence of virulence genes was determined by PCR method. Antibiotic susceptibility results revealed that the isolates were mostly susceptible to amikacin (61%), ceftazidime (60%), and imipenem (55%). Moreover, 59% of the isolates were multi-drug resistance (MDR). The most prevalent MDR pattern was aminoglycosides-penicillins-fluoroquinolones-carbapenems (15%). The presence of exoT, exoY, exoS and exoU genes was detected in 100%, 100%, 59%, and 41% of the tested isolates, respectively. Results points out the pattern of MDR and genetic diversity of type III secretion system among P. aeruginosa strains isolated from the burn population. Overall, the association of MDR and the presence of the specific virulence genes can be a predictive marker for the persistence of these isolates in the hospitals and subsequently a worse clinical condition for the affected patients.


Assuntos
ADP Ribose Transferases/genética , Antibacterianos , Proteínas de Bactérias/genética , Toxinas Bacterianas/genética , Unidades de Queimados , Queimaduras/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Proteínas Ativadoras de GTPase/genética , Glucosiltransferases/genética , Pseudomonas aeruginosa/genética , Sistemas de Secreção Tipo III , Fatores de Virulência/genética , Estudos Transversais , Humanos , Irã (Geográfico) , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/isolamento & purificação
3.
Mol Biol Rep ; 46(1): 207-215, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30415442

RESUMO

Recently, much attention has been focused on the use of miRNAs in cancer treatment. The role of proto-oncogene Janus kinase-2 (JAK-2) in proliferation and survival of gastric cancer has been previously documented. The aim of this study was to evaluate the effect of a chimera consisted of nucleolin specific aptamer (NCL-Apt) and miRNA let-7d on JAK2 expression level and activity in gastric cancer cells. NCL-Apt-miRNA let-7d chimera was prepared by two methods. Gastric cancer (MKN-45) cell line and control cell line of human dermal fibroblast (HDF) were treated with the chimera and the changes in JAK2 expression and activity were determined using real-time PCR and ELISA techniques, respectively. In MKN-45 cells, the chimera caused significant decrease in JAK2 expression level and activity compared to the aptamer alone and miRNA mimic negative control. Nevertheless, transfected miRNA let-7d showed remarkable reduction in the expression level of JAK2 in comparison with control state in both MKN-45 and HDF, confirmed unspecific effect of let-7d on normal and cancerous cells. With regard to the synergic effect of this chimera on JAK2 activity, it might be viewed as a therapeutic candidate in gastric cancer. However, further studies are warranted to prove it.


Assuntos
MicroRNAs/genética , Fosfoproteínas/fisiologia , Proteínas de Ligação a RNA/fisiologia , Neoplasias Gástricas/genética , Aptâmeros de Nucleotídeos/genética , Aptâmeros de Nucleotídeos/farmacologia , Linhagem Celular Tumoral , Quimera , Regulação da Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Janus Quinase 2/genética , Janus Quinase 2/fisiologia , MicroRNAs/fisiologia , Fosfoproteínas/genética , Dados Preliminares , Proteínas de Ligação a RNA/genética
4.
Iran J Pharm Res ; 17(4): 1537-1549, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30568710

RESUMO

miRNAs as one of the potential therapeutic agents have been recently considered for cancer treatment. AS1411 (aptNCL) is a DNA aptamer specifically binding to nucleolin protein on the cancer cell surface with antiproliferative effect. The aim of the study was to develop a conjugate consisting of aptNCL (as targeted delivery of therapeutic agent) and miRNA let-7d (as a tumor suppressor) using two different linking methods and also to evaluate the potential effect of the conjugates on the proliferation of gastric cancer (MKN-45) cell line compared to negative control cell line of human dermal fibroblast (HDF). Conjugation was performed covalently by SM (PEG)2 as a bifunctional crosslinker (conjugate-1) and noncovalently, using 19bp complementary sticky end sequences (conjugate-2). Nucleolin positive MKN-45 and nucleolin negative HDF cells were cultured and treated with the conjugates. Then, the changes in let-7d expression and cell proliferation were determined using Real-time PCR and MTT methods, respectively. In MKN-45 cells, the conjugates caused significant increase in let7-d uptake compared with HDF cells (P = 0.0001). The conjugate-1, likely due to its higher stability compared with the conjugate-2, led to significantly more increase in intracellular let-7d in MKN-45 cells (30 fold versus 15 fold, respectively, P = 0.0001). The conjugates revealed more potent antiproliferative effect against gastric cancer cells compared with aptNCL alone (P = 0.0001). It was found that the aptNCL-let-7d conjugate efficiently carried let-7d into the cancer cells. Also, it appears that in the setting of aptNCL-let-7d conjugate, let-7d and aptNCL moieties could cooperate and synergistically exhibit the antiproliferative effect on cancer cells.

5.
Avicenna J Med Biotechnol ; 10(1): 34-40, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29296265

RESUMO

Background: Type 4 pili (T4P) is an important virulence factor of Pseudomonas aeruginosa (P. aeruginosa). T4P pass the outer membrane through a large oligomeric channel made of a single PilQ protein that is most highly conserved at their C-termini. To develop a functional vaccine that can be used in clinical application, the secretin domain of the PilQ (PilQ380-706) was produced as a recombinant protein. Methods: A 981 bp fragment of C-terminal of the pilQ secretin (pilQ1138-2118) from was designed into the prokaryotic expression vector pET28a. The presence of the pilQ1138-2118 gene in the recombinant construct (pET28a/pilQ) was assessed by double digestion and PCR. After transformation, expression of the recombinant PilQ was induced by addition of IPTG. The expressed recombinant protein was purified by a modified method using a HisTrap affinity column and finally confirmed by SDS-PAGE. The functional activities of the produced PilQ380-706 confirmed by Western blot analysis and twitching inhibition assay. Results: The PCR and enzymatic digestion results showed the presence of the pilQ1138-2118 gene in the construct. The protein electrophoresis showed that the molecular weight of the recombinant PilQ380-706 is approximately 37 kDa. The Western blot analysis confirmed the specificity of specific IgG against the PilQ380-706 protein. The PilQ380-706 protein showed high biological activity in all of these standard assays. Conclusion: Since, the PilQ380-706 protein plays an important role in the biogenesis of pili; and thus, the primary establishment of P. aeruginosa; it seems that it can be used as a candidate vaccine or an adjuvant in the future studies.

6.
J Infect Dev Ctries ; 12(2): 109-114, 2018 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-31825912

RESUMO

INTRODUCTION: Leptospirosis is a widespread zoonotic disease which is endemic in Guilan province, Iran. Besides economic losses in the dairy industry, leptospirosis is also considered an important public health problem. This study aimed to evaluate two serological techniques, MAT and IgM-ELISA for detection of leptospiral antibodies. METHODOLOGY: A total of 185 samples were collected from individuals in Guilan province suspected of having leptospirosis from April 2016 to December 2016. Sera from participants were analyzed for Leptospira IgM antibodies using an available ELISA test and the MAT method. The specificity and sensitivity of the tests were calculated and compared. RESULTS: Of the 185 serum samples examined 114 (61.6%) and 94 (50.8%) samples were determined to be positive by MAT and IgM-ELISA, respectively. The results also showed that 17.5% of the sera that reacted positive in MAT were negative by IgM-ELISA, and 20.2% of IgM-ELISA positive sera were negative by MAT. We also showed that the MAT had specificity and sensitivity of 100%, when compared to leptospirosis-positive and negative serum samples. The specificity and sensitivity of IgM-ELISA was calculated as 78.8% and 82.4% respectively when compared with MAT. Bivariate analysis showed high correlation between the season, community of residence, possible reasons of pollution and leptospirosis (P < 0.1). CONCLUSION: Rural areas of Guilan, especially rice farming areas, are endemic for leptospirosis. Rice farmers have a high risk of infection with leptospirosis; infection is associated with direct exposure to rodent urine, gender (male) and season (spring).

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