Exogenous coenzyme Q10 modulates MMP-2 activity in MCF-7 cell line as a breast cancer cellular model

<p>Abstract</p> <p>Background/Aims</p> <p>Matrix Metalloproteinases 2 is a key molecule in cellular invasion and metastasis. Mitochondrial ROS has been established as a mediator of MMP activity. Coenzyme Q₁₀contributes to intracellular ROS regulation. Coenzyme Q₁₀beneficial effects on cancer are still in controversy but there are indications of Coenzyme Q₁₀complementing effect on tamoxifen receiving breast cancer patients.</p> <p>Methods</p> <p>In this study we aimed to investigate the correlation of the effects of co-incubation of coenzyme Q10 and N-acetyl-L-cysteine (NAC) on intracellular H2O2 content and Matrix Metalloproteinase 2 (MMP-2) activity in MCF-7 cell line.</p> <p>Results and Discussion</p> <p>Our experiment was designed to assess the effect in a time and dose related manner. Gelatin zymography and Flowcytometric measurement of H2O2 by 2'7',-dichlorofluorescin-diacetate probe were employed. The results showed that both coenzyme Q10 and N-acetyl-L-cysteine reduce MMP-2 activity along with the pro-oxidant capacity of the MCF-7 cell in a dose proportionate manner.</p> <p>Conclusions</p> <p>Collectively, the present study highlights the significance of Coenzyme Q₁₀effect on the cell invasion/metastasis effecter molecules.</p

Collagen-Binding Peptidoglycans Inhibit MMP Mediated Collagen Degradation and Reduce Dermal Scarring

Scarring of the skin is a large unmet clinical problem that is of high patient concern and impact. Wound healing is complex and involves numerous pathways that are highly orchestrated, leaving the skin sealed, but with abnormal organization and composition of tissue components, namely collagen and proteoglycans, that are then remodeled over time. To improve healing and reduce or eliminate scarring, more rapid restoration of healthy tissue composition and organization offers a unique approach for development of new therapeutics. A synthetic collagen-binding peptidoglycan has been developed that inhibits matrix metalloproteinase-1 and 13 (MMP-1 and MMP-13) mediated collagen degradation. We investigated the synthetic peptidoglycan in a rat incisional model in which a single dose was delivered in a hyaluronic acid (HA) vehicle at the time of surgery prior to wound closure. The peptidoglycan treatment resulted in a significant reduction in scar tissue at 21 days as measured by histology and visual analysis. Improved collagen architecture of the treated wounds was demonstrated by increased tensile strength and transmission electron microscopy (TEM) analysis of collagen fibril diameters compared to untreated and HA controls. The peptidoglycan's mechanism of action includes masking existing collagen and inhibiting MMP-mediated collagen degradation while modulating collagen organization. The peptidoglycan can be synthesized at low cost with unique design control, and together with demonstrated preclinical efficacy in reducing scarring, warrants further investigation for dermal wound healing

Distribution of ِHuman Leptospirosis in Guilan Province, Northern Iran

Background: Leptospirosis is the most widespread zoonosis in the world and is more prevalent in tropical and temperate regions. Guilan Province in north of Iran, is an endemic region of human leptospirosis. Since diagnosis of leptospirosis according to clinical symptoms is very difficult due to lack of characteristic pathogonomic sign(s), laboratory support is necessary. Methods: In 2003, we obtained blood samples from patients hospitalized in main general hospitals of Guilan Province and were suspected as having leptospirosis according to their clinical presentations. We examined 995 sera by a commercial IgM and IgG ELISA kit to find positive cases. Results: 62.7% of positive cases were male and about 86% of them were farmer. High distribution rate of leptospirosis was seen in middle-aged people (65% in 20-50 years old). Conclusion: It seems that leptospirosis has a high occurrence in major cities and is mostly distributed in warm months of the year. Demographic analysis of the results indicates that leptospirosis is typically a rural and an occupational disease in the area

The Effects of Arsenic Trioxide and Zoledronic Acid on Malignant Plasma Cells Derived from Bone Marrow Cells of Multiple Myeloma Patients

&quot;nBackground: Multiple myeloma (MM) is a disease of plasma cells that has fatal consequences. New agents associated with mo&amp;shy;lecular targets have prompted clinical investigators to design new treatment strategies initially for advanced MM and later for newly diagnosed MM, with encouraging preliminary results. &amp;nbsp;We devised a project to assess the mechanisms of ac&amp;shy;tion of two drugs, Ar&amp;shy;senic trioxide (ATO) and Zoledronic acid (Zometa) on Bone marrow mononuclear cells (BMMCs) de&amp;shy;rived from patients.&quot;nMethods: Bone marrow samples were collected from 10 patients after receipt of formal consent. BMMCs were collected from samples. In two parallel sets of experiments, BMMCs were treated with 0.5, 2, 6 &amp;micro;M ATO and 0.1, 10, 100 &amp;micro;M Zo&amp;shy;meta, for 72 h. The following analyses were then performed on treated cells as compared to untreated cells (assumed as con&amp;shy;trol): cytotoxicity using Micro culture tetrazolium test (MTT assay); matrix metalloproteinase-2 zymography; comparative gene expression analysis of IL-6, vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-l).&quot;nResults: MTT assay showed significant proliferation inhibition in ATO high dose treatment (6 uM). However, no signifi&amp;shy;cant in&amp;shy;hibitory effect of Zometa was seen. Zymography analyses showed significant decrease in gelatinolytic activity in treated cells. Analyses of gene expression using Real-Time RT-PCR methodology showed significant decrease in IL-6, ICAM-1, and VEGF genes as normalized against Hypoxanthine phosphoribosyltransferase normalizer and as compared with untreated cells.&quot;nConclusion: Both ATO and Zometa could significantly decrease MM cells critical phenotype and genotype. This finding could support the hypothesis that ATO or Zometa could inhibit growth and metastasis of malignant cells

"A PCR-RFLP Method to Identification of the Important Opportunistic Fungi: Candida Species, Cryptococcus neoformans, Aspergillus famigatus and Fusarium solani"

Deep-seated fungal infection present with non specific symptoms and involove a large number of different organisms. DNA-based technology offers for eariler detection of fungal pathogens and then earlier initiation of antifungal therapy. In this study universal primers common to almost all fungi were used to amplification of internal transcribe spacer 1 and 2 region. Subsequent restriction enzyme analysis of PCR products, using HpaII allows us to identify the most medically opportunistic important fungi: Candida albicans, C. glabrata, C. tropicalis, C. kruzei, C. guilliermondi, Cryptococcus neoformans, Aspergillus fumigatus and Fusarium solani, according to sizely different bands in polyacrilamid gel electrophoresis. It seems that this panel of PCR-RFLP could be a rapid and useful molecular approach in diagnostic studies of invasive opportunistic fungal infections

"Attempt to Detect Mycoplasma and Chlamydia by Culture in the Synovial Fluid of Patients with Arthritis"

The objective was to investigate the presence of Mycoplasma and Chlamydia in the synovial fluid of patients with rheumatoid arthritis (RA) and other chronic arthritides. Samples of synovial fluid (SF) were collected from all patients presenting with an articular effusion. Seventy SF samples were subjected to study for Mycoplasma and cultured on standard media for Mycoplasma. The 70 other SF samples were subjected to study for the presence of Chlamydia and cultivated on cell cultures specially MC coy cell lines. All standard cultures for Mycoplasma and Chlamydia remained negative, consistent with the fact that synovial fluid is sterile, despite of many investigations that have indicated DNA of some bacteria in the SF of patients with arthritis. However, as many other attempts to detect the presence of these fastidious organisms in the joints of patients with such arthritides have failed, the question of their possible roll in the pathogenesis of human rheumatic diseases remains controversial and needs to be re-examined

Fabrication of Docetaxel Surfaced Fe3O4 Magnetite Nanoparticles and their Cytotoxicity on 4 T1 Breast Cancer Cells

Background:In the recent years, there is an increasing attention to the using of Fe3O4 magnetite nanoparticles (MNPs) as drug delivery systems. Application of this nanoparticles could profit advantages of nanomedicine to enhance biological activity of pharmaceutical ingredients. Methods:Fe3O4 MNPs were synthesised by a chemical method and characterized by transmission electron microscopy and energy-dispersive spectroscopy techniques. In the next step, docetaxel-coated Fe3O4 MNPs were prepared, using percipitation method. The surface chemistry of docetaxel-coated Fe3O4 MNPs as well as their thermal decomposition characteristics were examined using fourier transform infrared spectroscopy and thermogravimetric analyzer equipment, respectively. The cytotoxicity assay was conducted on 4 T1 breast cancer carsinoma by MTT assay to evaluate the possible in vitro antiproliferative effects of docetaxel-coated Fe3O4 MNPs. Results:During precipitation process, docetaxel molecules were precipitated on the surface of Fe3O4 MNPs by the ratio of 3:100 w/w which indicates that each milligram of coated Fe3O4 MNPs averagely contained 30 mug pure docetaxel compound. Docetaxel showed aniproliferative effects against mentioned cell line. The higestest concentartion of docetaxel (80 mug/ml) caused about 80% cell death. However, the results demostarted that much lower amounts of docetaxel will be needed in combination of Fe3O4 MNPs to produce the potent antiproliferative effect compared to docetaxel alone. Dose response cytotoxicity assay of docetaxel-coated Fe3O4 MNPs against 4 T1 breast cancer cells showed that lower amount of docetaxel (0.6 mug/ml) can exhibit higher cytotoxic effect against this cancer cell line (90% cell death)

Effect of Tindurin on Immunopathogenesis Mechanism of Collagen-Induced Arthritis

Rheumatoid arthritis is a chronic inflammatory disease characterized by the sequestration of various leukocyte subpopulations within both the developing pannus and synovial space. This study was undertaken to examine the therapeutic potency of tindurin in experimental rheumatoid arthritis. Collagen-induced arthritis (CIA) was induced by intradermally immunization of Lewis rats at the base of the tail. The paws and knees were then removed for histopathology and radiography analysis. Using fibrosarcoma cell line the apoptosis process was measured by Terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method. Our data showed that the i.p. injection of tindurin to arthritic rats induced a significant reduction in paw edema. Histopathological assessment showed reduced inflammatory cells infiltrate, tissue edema and bone erosion in joints of treated rats. Moreover, our results in radiography were in line with histological findings as well as tindurin was found to induce apoptosis of treated cells in comparison with positive, negative and non-treated ones. Our findings revealed the therapeutic effect of tindurin in experimental model of rheumatoid arthritis in comparison with methotrexate as a choice drug

Assessment of Ail Gene Marker Amplicon for Mo­lecular Characterization of Pathogenic Yersinia enterocolitica in Food Samples Collected in Iran

Background: To assess the utility of the chromosomal ail virulence gene sequence for detection of pathogenic Yersinia en&amp;shy;terocolitica in raw meet food products (beef, lamb, and chicken). Methods: This study included 39 Yersinia enterocolitica positive cultures from suspicious food samples, in a working pe&amp;shy;riod of six months. These samples were referred to the &amp;quot;Food-Borne Diseases and Chronic Diarrhea Lab at Research Cen&amp;shy;tre for Gastric and Liver Diseases&amp;quot; of the Taleghani Hospital at Shahid Beheshti University of Medical Sciences, Te&amp;shy;hran, Iran. Isolates from 8 cultured Y. intermedia, Y. aldovi, Y. intermedia type O:45, Y. kristensenii, Y. enterocolitica type O:12/26, Y. enterocolitica type1/7/8, Y. frederiksenii type O:39, and Y. enterocolitica type O:8 samples were in&amp;shy;cluded in the study. Four non-Yersinia species Salmonella typhi, Shigella dysenteriae, Shigella flexeneri, and Proteus mirabi&amp;shy;lis were used for specificity testing. An established Yersinia type O:9 was used as positive control and for sensitiv&amp;shy;ity testing. An in-house real-time PCR assay was designed in order to rapidly and specifically identifies the pres&amp;shy;ence of specific Yersinia species. Results: Out of 39 tested Y. enterocolitica samples, 6(2.3%) showed positive results for the ail gene PCR prod&amp;shy;uct, typed as O:8, and O:9, respectively. PCR products were sent for sequencing. Two sequences were registered with the Na&amp;shy;tional Center for Biotechnology Information (NCBI Genbank) as polymorphic ail gene sequences under the acces&amp;shy;sion numbers of DQ157767 and DQ003329. Conclusions: Collectively, this test is well adapted for definite confirmation of pathogenic Y. en&amp;shy;terocolitica in food sam&amp;shy;ples