scholarly journals The dual inhibitor of lipoxygenase and cyclooxygenase ML3000 decreases the expression of CXCR3 ligands

2007 ◽  
Vol 67 (4) ◽  
pp. 524-529 ◽  
Author(s):  
C Ospelt ◽  
M Kurowska-Stolarska ◽  
M Neidhart ◽  
B A Michel ◽  
R E Gay ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Polymerase Chain Reaction ◽  
Molecular Mechanisms ◽  
Synovial Fibroblasts ◽  
Target Cells ◽  
Chain Reaction ◽  
Dual Inhibitor ◽  
Protein Levels ◽  
Polymerase Chain ◽  
The Impact

Objective:To find previously unknown properties of ML3000, a competitive inhibitor of the cyclooxygenase and the lipoxygenase (LO) pathway.Methods:Gene expression of ML3000 treated and untreated rheumatoid arthritis synovial fibroblasts were measured with Affymetrix gene arrays. Downregulation of chemokine (C-X-C motif) ligands CXCL9, CXCL10 and CXCL11 was verified with Real-time polymerase chain reaction, CXCL10 protein levels were determined with ELISA. Rheumatoid arthritis synovial fibroblasts were treated with the cyclooxygenase inhibitor naproxen, the 5-LO inhibitor BWA4C and the 5-lipoxygenase-activating protein (FLAP) inhibitor MK886, and consecutive changes in CXCL10 protein levels measured. 5-LO expression was determined by polymerase chain reaction and Western blot.Results:In synovial fibroblasts and monocyte-derived macrophages ML3000 inhibited the tumour necrosis factor induced expression of CXCL9, CXCL10 and CXCL11, which are all ligands of the chemokine receptor CXCR3. No effect was observed in monocytes. Whereas inhibition of the cyclooxygenase pathway or the FLAP protein showed no effect, blockade of 5-LO significantly downregulated CXCL10 protein levels. 5-LO mRNA was detected in monocytes and in monocyte-derived macrophages. All tested cell types expressed 5-LO protein.Conclusions:ML3000 effectively downregulates CXCR3 ligands. This study confirms that a thorough analysis of the impact of a drug on its target cells cannot only reveal unexpected properties of a substance, but also helps to understand the underlying molecular mechanisms. Accordingly, our data provide the basis for further clinical studies testing the application of ML3000 in diseases such as rheumatoid arthritis or multiple sclerosis.

scholarly journals Macrophage migration inhibitory factor may play a protective role in osteoarthritis

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Ming Liu ◽  
Zikun Xie ◽  
Guang Sun ◽  
Liujun Chen ◽  
Dake Qi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Migration Inhibitory Factor ◽  
Macrophage Migration Inhibitory Factor ◽  
Protective Role ◽  
Macrophage Migration ◽  
Chain Reaction ◽  
Protein Levels ◽  
Tnf Α ◽  
Polymerase Chain

Abstract Background Osteoarthritis (OA) is the most prevalent form of arthritis and the major cause of disability and overall diminution of quality of life in the elderly population. Currently there is no cure for OA, partly due to the large gaps in our understanding of its underlying molecular and cellular mechanisms. Macrophage migration inhibitory factor (MIF) is a procytokine that mediates pleiotropic inflammatory effects in inflammatory diseases such as rheumatoid arthritis (RA) and ankylosing spondylitis (AS). However, data on the role of MIF in OA is limited with conflicting results. We undertook this study to investigate the role of MIF in OA by examining MIF genotype, mRNA expression, and protein levels in the Newfoundland Osteoarthritis Study. Methods One hundred nineteen end-stage knee/hip OA patients, 16 RA patients, and 113 healthy controls were included in the study. Two polymorphisms in the MIF gene, rs755622, and -794 CATT5-8, were genotyped using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and PCR followed by automated capillary electrophoresis, respectively. MIF mRNA levels in articular cartilage and subchondral bone were measured by quantitative polymerase chain reaction. Plasma concentrations of MIF, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) were measured by enzyme-linked immunosorbent assay. Results rs755622 and -794 CATT5-8 genotypes were not associated with MIF mRNA or protein levels or OA (all p ≥ 0.19). MIF mRNA level in cartilage was lower in OA patients than in controls (p = 0.028) and RA patients (p = 0.004), while the levels in bone were comparable between OA patients and controls (p = 0.165). MIF protein level in plasma was lower in OA patients than in controls (p = 3.01 × 10−10), while the levels of TNF-α, IL-6 and IL-1β in plasma were all significantly higher in OA patients than in controls (all p ≤ 0.0007). Multivariable logistic regression showed lower MIF and higher IL-1β protein levels in plasma were independently associated with OA (OR per SD increase = 0.10 and 8.08; 95% CI = 0.04–0.19 and 4.42–16.82, respectively), but TNF-α and IL-6 became non-significant. Conclusions Reduced MIF mRNA and protein expression in OA patients suggested MIF might have a protective role in OA and could serve as a biomarker to differentiate OA from other joint disorders.

Detection of Aspergillus fumigatus by Polymerase Chain Reaction (PCR)

2019 ◽  
Vol 10 (04) ◽  
pp. 655-661
Author(s):  
Zainab H Abood AL-Asadi
Keyword(s):  
Polymerase Chain Reaction ◽  
Aspergillus Fumigatus ◽  
Oligonucleotide Primer ◽  
Rrna Gene ◽  
Fungal Culture ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Internal Transcribed Spacer Region ◽  
Polymerase Chain ◽  
The Impact

Aspergillosis refers to fungi infections of the respiratory tract caused by Aspergillus species, especially Aspergillus fumigatus. Infection of A. fumigatus was increased in the last few years due to either resistances to antibiotics or the influence of other factors such as other fungal infections. The present study aimed to review the impact of Aspergillus fumigatus in Aspergillosis cases, and study the role of Singleplex PCR for amplification of ITS1, ITS4 of rRNA gene in the detection of fungal isolate. In this study, One hundred sputum samples were collected from patients admitted to the specialize chest and respiratory diseases center / Baghdad who were suffering from respiratory problems. During these studied, molds were isolation and identification based on Conventional method (Direct microscopy by using 10% KOH, and fungal culture was done on Sabouraud Dextrose agar supplemented with chloramphenicol and on Czapek-Dox agar incubated at 37°C and examined for 3-7 days then macroscopic, microscopic examination of the colony by(lactophenol cotton blue stain )and molecular methods by using Polymerase chain reaction (PCR)technique for identification. The 10% KOH examination was positive for 35 cases, while laboratory culturing was positive for 53 cases. Aspergillus sp were isolated from 44(83%) patients; A. fumigatus was isolated in 23 (42. 4%) patients while A. flavus, A. niger, and A. terreus were isolated from 11 (20. 08%), (13. 2%) and 3 (5. 7%) patients respectively, also isolated Penicillium spp. at percentage 1(1. 9%). In this study. The ages of participants ranged from 10-70years with a mean age of 34years, the males were more susceptible to fungal infection, were recorded 35/53 (66. 3), compared to females were 18/53 (33. 96). The infection of fungi was more prevalent in ages 30-40recorded 26(53. 06%) followed by ages 40-50, 13(26. 5), while the lowest infection recorded in the age group 10- 20 years was 2(2. 04%). DNA isolated from twenty-three A. fumigatus isolates was used as a template, and the specific of oligonucleotide primer sequences were used in conventional PCR to detect the presence of internal transcribed spacer region ( ITS) region of the rRNA gene for Aspergillus fumigates. The results of the PCR amplification of the rRNA gene showed that this gene was present in 19 samples out 23 positive samples which isolation with a PCR product size of approximated 385 bp, while 4 samples out 23 positive samples showed negative results for the presence of this gene as indicated by the absence of the PCR products in their relevant lanes. Statistical analysis revealed that the PCR to have a sensitivity of 95. 1% in the detection of Aspergillus fumigatus in Aspergillosis cases. Polymerase chain reaction (PCR) is a rapid, specific, and sensitive method to detect Aspergillus fumigatus in aspergillosis cases of humans.

scholarly journals The impact of COL1A1 and COL6A1 expression on hypospadias and penile curvature severity

BMC Urology ◽  
2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Prahara Yuri ◽  
Gunadi ◽  
Rahmadani Puji Lestari ◽  
Firly Putri Fardilla ◽  
Wiwit Ananda Wahyu Setyaningsih ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Polymerase Chain Reaction ◽  
External Genitalia ◽  
Penile Curvature ◽  
Moderate Correlation ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Distal Hypospadias ◽  
Male External Genitalia ◽  
The Impact

Abstract Background Hypospadias, the most frequent congenital male external genitalia abnormality, is usually associated with curvature of the ventral penis, i.e. chordee. Abnormality of darto tissue has been suggested as the pathophysiology of chordees. Collagen is one of the most abundant fibrous proteins within the extracellular matrix. In this study, we determined the expression of collagen 1 (COL1A1) and COL6A1 in patients with hypospadias and associated them with the severity of penile curvature. Methods We included 60 children < 18 years old, consisting of 20 distal hypospadias, 20 proximal hypospadias patients, and 20 controls in our institution from 2017 – 2020. The expression of COL1A1 and COL6A1 in darto tissue was determined by reverse-transcriptase polymerase chain reaction (qPCR). The penile curvature severity was classified as mild (< 30 degrees), moderate (30–60 degrees), and severe (> 60 degrees). Results qPCR showed that COL1A1 and COL6A1 expression was significantly downregulated in the distal (0.88 (0.38–2.53) and 0.54 (0.16–4.35), respectively) and proximal 0.76 (0.33–2.57) and 0.57 (0.18–1.38), respectively) hypospadias groups compared to controls (1.85 (0.24–4.61) and 0.93 (0.17–4.06), respectively) with p-values of 0.024 and 0.018, respectively. Furthermore, there was a moderate correlation between COL1A1 and COL6A1 expression (r = 0.458, p < 0.0001). Interestingly, COL1A1 and COL6A1 were also significantly downregulated in the moderate and severe chordee groups compared to the mild chordee groups, with p-values of 0.003 and 0.037, respectively. Conclusions Aberrant COL1A1 and COL6A1 expression might affect abnormalities in darto tissue and penile curvature severity in hypospadias patients.

scholarly journals Ligation-Mediated Polymerase Chain Reaction Detection of 8-Oxo-7,8-Dihydro-2′-Deoxyguanosine and 5-Hydroxycytosine at the Codon 176 of the p53 Gene of Hepatitis C-Associated Hepatocellular Carcinoma Patients

2020 ◽  
Vol 21 (18) ◽  
pp. 6753
Author(s):  
Andrea Galli ◽  
Armelle Munnia ◽  
Filippo Cellai ◽  
Mirko Tarocchi ◽  
Elisabetta Ceni ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Polymerase Chain Reaction ◽  
Hepatitis C ◽  
Molecular Mechanisms ◽  
Active Oxygen Species ◽  
Dietary Exposure ◽  
P53 Gene ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Polymerase Chain

Molecular mechanisms underlying Hepatitis C virus (HCV)-associated hepatocellular carcinoma (HCC) pathogenesis are still unclear. Therefore, we analyzed the levels of 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) and other oxidative lesions at codon 176 of the p53 gene, as well as the generation of 3-(2-deoxy-β-d-erythro-pentafuranosyl)pyrimido[1,2-α]purin-10(3H)-one deoxyguanosine (M1dG), in a cohort of HCV-related HCC patients from Italy. Detection of 8-oxodG and 5-hydroxycytosine (5-OHC) was performed by ligation mediated-polymerase chain reaction assay, whereas the levels of M1dG were measured by chromatography and mass-spectrometry. Results indicated a significant 130% excess of 8-oxodG at –TGC– position of p53 codon 176 in HCV-HCC cases as compared to controls, after correction for age and gender, whereas a not significant increment of 5-OHC at –TGC– position was found. Then, regression models showed an 87% significant excess of M1dG in HCV-HCC cases relative to controls. Our study provides evidence that increased adduct binding does not occur randomly on the sequence of the p53 gene but at specific sequence context in HCV-HCC patients. By-products of lipid peroxidation could also yield a role in HCV-HCC development. Results emphasize the importance of active oxygen species in inducing nucleotide lesions at a p53 mutational hotspot in HCV-HCC patients living in geographical areas without dietary exposure to aflatoxin B1.

Mo1852 The Impact of Polymerase Chain Reaction on Clostridium difficile Detection and Clinical Outcomes

2015 ◽  
Vol 148 (4) ◽  
pp. S-726-S-727
Author(s):  
Mona Akbari ◽  
Victor Novack ◽  
Ciaran P. Kelly ◽  
Daniel A. Leffler
Keyword(s):  
Polymerase Chain Reaction ◽  
Clostridium Difficile ◽  
Clinical Outcomes ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
The Impact

scholarly journals Analysis of the impact of long non-coding RNA gene polymorphisms (ANRIL, MALAT1, HOTAIR) on some clinical and pathologic features in patients with bladder cancer

2021 ◽  
Vol 25 (2(98)) ◽  
pp. 13-21
Author(s):  
A. Volkohon ◽  
V. Harbuzova ◽  
I. Danilishin ◽  
D. Nechyporenko ◽  
O. Ataman
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Tumor Size ◽  
Transitional Cell ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Blood Hemoglobin ◽  
Pathologic Features ◽  
Polymerase Chain ◽  
The Impact

Objective – to analyze the possible association between the SNPs rs4977574 (ANRIL gene), rs3200401 (MALAT1 gene), rs1899663 (HOTAIR gene) and tumor size and some clinical and laboratory testing data in patients with transitional cell carcinoma of the urinary bladder (TCCUB).Material and methods. Whole venous blood of 141 patients with TCCUB was used. The genotyping of rs4977574 and rs3200401 sites was performed by real-time polymerase chain reaction (Real-time PCR). The rs1899663 locus genotyping was done by polymerase chain reaction followed by restriction fragment length polymorphism analysis (PCR-RFLP). Mathematical analysis of the obtained data was performed using the software package SPSS (version 17.0).Results. It was found that individuals with TT-genotype (rs3200401-polymorphism) have a lower blood hemoglobin content ((106.3 ± 23.9) g/l; P = 0.043) and higher blood glucose ((7.1 ± 2.3) mmol/l; P = 0.043) and creatinine ((104.5 ± 33.8) μmol/l; P = 0.022) than patients with CC genotype (respectively: (131.1 ± 21.9) g/l); (5.4 ± 1.5) mmol/l); (83.8 ± 18.5) μmol/l)). TT-homozygotes also have the higher tumor width ((4.2 ± 1.7) cm; P = 0.027) than in CC-homozygotes ((2.9 ± 1.1) cm). No significant association between rs4977574, rs1899663 loci and studied parameters was found.Conclusion. The MALAT1 gene rs3200401 polymorphism is associated with tumor size and blood hemoglobin, glucose, and creatinine levels in patients with TCCUB. No association between rs1899663, rs4977574 loci and clinical and pathological features in patients with TCUUB was detected.

Sign in / Sign up

Export Citation Format

Share Document