Development of a rapid and accurate quantification method for the detection of microRNAs (miRNAs) has been desired, in particular, when they are differently expressed in normal and pathological conditions. However, various methods for the quantification of small non-coding RNAs as well as miRNAs have been described. These methods mainly include hybridization-based approaches such as primer extension, northern blotting, microarray profiling, and reverse transcription (RT) PCR. Here, we developed a simple and rapid method based on stem-loop primer-based real-time PCR assay for sensitive and accurate detection of mature miRNAs. Initially, a miRNA-specific stem-loop RT primer is used for RT, which is followed by TaqMan real-time PCR assay using specific forward primer in combination with universal reverse primer and TaqMan probe. The assay has shown high sensitivity (≤50 copies/reaction) for miRNA detection in two breast cancer cell lines, MCF-7 and MDA-MB-231. This assay might be implicated as a rapid and cost effective method for the detection of small non-coding RNAs.
The study showed that the consumption of salt among the Iranian population is higher than the level recommended by WHO. To reduce salt intake, it is necessary to adopt a combination of nationwide policies such as food reformulation and food labelling.
The therapeutic potential of bone marrow mesenchymal stem cells (MSCs) in kidney failure has been examined in some studies. However, recent findings indicate that after transplantation, these cells home to kidneys at very low levels. Interaction of stromal derived factor-1 (SDF-1) with its receptor, CXCR4, is of pivotal importance in migration and homing. Recently, CXCR7 has also been recognized as another SDF-1 receptor that interacts with CXCR4 and modulates its functions. In this study, CXCR4 and CXCR7 were separately and simultaneously overexpressed in BALB/c bone marrow MSCs by using a lentiviral vector system and the homing and renoprotective potentials of these cells were evaluated in a mouse model of cisplatin-induced acute kidney injury. Using flow cytometry, immunohistochemistry, and real-time PCR methods for detection of GFP-labeled MSCs, we found that although considerably entrapped in lungs, native MSCs home very rarely to kidneys and bone marrow and this rate cannot be significantly affected by CXCR4 and/or CXCR7 upregulation. Transplantation of neither native nor genetically engineered MSCs ameliorated kidney failure. We concluded that overexpression of CXCR4 and CXCR7 receptors in murine MSCs cannot improve the homing and therapeutic potentials of these cells and it can be due to severe chromosomal abnormalities that these cells bear during ex vivo expansion.
This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
Methicillin-Resistant Staphylococcus aureus (MRSA) is responsible for an increasing number of serious hospital and community acquired infections. Virulence gene expression in Staphylococcus aureus is orchestrated by regulators such as the accessory gene regulator (agr). Staphylococcal strains are divided into four major agr groups (agrI-IV) on the basis of agrD and agrC polymorphisms. The purpose of this study was to define the prevalence of MRSA strains in appointed Tehran's hospitals and then to define and compare the proportion of agr I, II, III, IV polymorphisms between MRSA and Methicillin Sensitive Staphylococcus aureus (MSSA) strains. A total of 235 isolates were evaluated by conventional antibiotic susceptibility tests and PCR for agr and mecA genes. 112 strains were MRSA (47.5%) and the most prevalent agr specific group was agr I followed by agr III, agr II and agr IV, respectively. The prevalence of agr groups amongst MRSA and MSSA strains was not statistically significant (P≥0.05). This study suggests that agr I is not only the most prevalent agr type in MRSAs but also the most common one in Methicillin Sensitive Staphylococcus aureus (MSSA) strains in Iran.
Background: HPV related cervical cancer as one of the most common women cancers in developing countries. Regarding accessibility of commercial vaccines, any long or short term modality for integrating preventive immunization against HPV in a national program needs comprehensive information about HPV prevalence and its genotypes. The important role of selecting most accurate diagnostic technologies for obtaining relevant data is underlined by different assays proposed in the literature. The main objective of the present study was to introduce an in-house HPV typing assay using multiplex real time PCR with reliable results and affordable cost for molecular epidemiology surveys and diagnosis. Materials and Methods
ResultsConclusions: According to acceptable performance, easy access to primers, probes and other consumables, affordable cost per test, this method can be used as a diagnostic assay in molecular laboratories and for further planning of cervical carcinoma prevention programs.
Keywords
Materials and Methods
Patients' SpecimensEmbedded Tissue (FFPE) specimens) from women with known cervical precancerous and cancer referred to Tehran to meet ethical considerations, each patient was informed about the objectives of study and a signed a consent form before entering the study. After cytologic examination of collected Liquid Based Cytology (LBC) and FFPE blocks C and FFPE tissue blocks in a proper storage area until experimental phase. 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68 containing partial length L1 sequence (20 e10 4 copy/ µl) were ® ,Germany, to be used for preparing standard samples necessary for evaluation studies.
HPVs standard plasmidsExtraction of HPV DNA Viral DNA from Thin Prep preservatives and 20-30 mg Germany), according to the manufacturer's instructions.
Design of primers and probesnucleotide). Analysis of extensive bioinformatics was done on primers and probes designed by Oligo7, Primer as internal control primers and probe were considered in inhibition and quality of extracted DNA.
In-house multiplex real time PCR for HPV genotypingas quencher dye at the 3' end of probes sequences. For carried out in a 25µl reaction volume containing 12.5µl 2Xand 5 µl DNA extracted. These reaction mixtures were Orange Channels). Furthermore, all of the four mixtures were tested against known positive specimens of HBV, dilutions of HPV plasmids were prepared from 1 to 1000 copy/ µl as separated and mixed samples.
ResultsThe quality and performance of developed assay was evaluated focusing on analytical sensitivity (detection negative and 20 HPV positive samples (both as single specimens and mixed cocktails with different copy numbers of each genotype per micro liter from 1 to 1000) signal in all samples.Analytical sensitivity high risks and low risks HPV genotypes) were tested and only one sample (genotype 52) was falsely negative.Detection limit prepared (from 1 to 1000 copy /µl) and the assay could detect 5 copy/µl of each HPV genotypes in single and mixed cocktails. Data for these evaluation...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.