Farid Azizi Jalilian

Chemical composition and antibacterial and cytotoxic activities of allium hirtifolium boiss

Allium hirtifolium Boiss. known as Persian shallot, is a spice used as a traditional medicine in Iran and, Mediterranean region. In this study, the chemical composition of the hydromethanolic extract of this plant was analyzed using GC/MS. The result showed that 9-hexadecenoic acid, 11,14-eicosadienoic acid, and n-hexadecanoic acid are the main constituents. The antibacterial activity of the shallot extract was also examined by disk diffusion and microdilution broth assays. It was demonstrated that Persian shallot hydromethanolic extract was effective against 10 different species of pathogenic bacteria including methicillin resistant Staphylococcus aureus (MRSA), methicillin sensitive Staphylococcus aureus (MSSA), Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Escherichia coli, Escherichia coli O157:H7, Salmonella typhimurium, Proteus mirabilis, and Klebsiella pneumoniae. Specifically, the minimum concentration of the extract which inhibited bacterial growth (MIC values) was 1.88 mg/mL for most of the gram-positive bacteria. This concentration was not much different from the concentration that was safe for mammalian cells (1.50 mg/mL) suggesting that the hydromethanolic extract of Persian shallot may be a safe and strong antibacterial agent.

Novel molecular, cytotoxical, and immunological study on promising and selective anticancer activity of Mung bean sprouts

BackgroundThe anticancer and immunomodulatory activity of mung bean sprouts (MBS) and the underlying mechanisms against human cervical and hepatocarcinoma cancer cells were explored.MethodsMBS cytotoxicity and MBS-induced anticancer cytokines, TNF-α and IFN-β from cancer cells, and immunological cytokines, IL-4, IFN-γ, and IL-10 from peripheral mononuclear cells (PMNC) were assessed by MTS and ELISA assays. Apoptotic cells were investigated by flow cytometry. The expression level of apoptotic genes (Bax, BCL-2, Capsases 7–9) and cell cycle regulatory genes (cyclin D, E, and A) and tumor suppressor proteins (p27, p21, and p53) was assessed by real-time qPCR in the cancer cells treated with extract IC50.ResultsThe cytotoxicity on normal human cells was significantly different from HeLa and HepG2 cells, 163.97 ± 5.73, 13.3 ± 0.89, and 14.04 ± 1.5 mg/ml, respectively. The selectivity index (SI) was 12.44 ± 0.83 for HeLa and 11.94 ± 1.2 for HepG2 cells. Increased levels of TNF-α and IFN-β were observed in the treated HeLa and HepG2 culture supernatants when compared with untreated cells. MBS extract was shown to be an immunopolarizing agent by inducing IFNγ and inhibiting IL-4 production by PBMC; this leads to triggering of CMI and cellular cytotoxicity. The extract induced apoptosis, in a dose and time dependent manner, in treated HeLa and HepG2, but not in untreated, cells (P < 0.05). The treatment significantly induced cell cycle arrest in G0/G1 in HeLa cells. The percentage of cells in G0/G1 phase of the treated HeLa cells increased from 62.87 ± 2.1%, in untreated cells, to 80.48 ± 2.97%. Interestingly, MBS IC50 induced the expression of apoptosis and tumor suppressor related genes in both HeLa and HepG2 cells. MBS extract succeeded in inducing cdk-inhibitors, p21, p53, and p27 in HeLa cells while it induced only p53 in HepG2 cells (P < 0.05). This is a clue for the cell type- specific interaction of the studied extract. These proteins inhibit the cyclin-cdk complexes apart from the presence of some other components that might stimulate some cyclins such as cyclin E, A, and D.ConclusionMBS extract was shown to be a potent anticancer agent granting new prospects of anticancer therapy using natural products.

Sulfonamide Resistance Genes (sul) M in Extended Spectrum Beta Lactamase (ESBL) and Non-ESBL Producing Escherichia coli Isolated From Iranian Hospitals.

Background: Extensive use of cotrimoxazole has been associated with increasing level of Escherichia coli resistance. Objectives: In the current study, we focused on assessing the prevalence of E. coli resistance to cotrimoxazole and frequency of its associated genes. Materials and Methods: One-hundred and forty-four E. coli isolates were identified during March 2007 to April 2012 at Ilam hospitals and Milad (Tehran) hospital. Antibiotic susceptibility for screening of resistance isolates was done by the Kirby-Bauer method. The sul1, sul2, sul3, dfrA1, dfrA5, int1, blaTEM, blaSHV and CTX-M genes were detected by polymerase chain reaction (PCR) amplification. Plasmid curing was done for identifying correlations between resistance genes and plasmids. Results: Amongst the 144 E. coli isolates, seventy-two (50%) Extended Spectrum Beta Lactamase (ESBL)-producing and seventy-two (50%) non-ESBL-producing E. coli isolates were identified; eighty-seven isolates (60.41%) were resistant to cotrimoxazole. Frequencies of sul1, sul2 and sul3, were 81% (116 isolates), 67% (96 isolates) and 2.29% (three isolates), respectively. Furthermore, 50.57% (72 isolates) had sul1 and sul2, 2.29% (3 isolates) contained sul2 and sul3, and 2.29% (three isolates) contained sul1, sul2 and sul3 genes, simultaneously. Thirty-four (39.1%) of the isolates had the dfrA1 gene. Five (5.7%) of the isolates had the dfrA5 gene. Sixty-eight (78.2%) strains contained the int1 gene. Furthermore, dfrA1 and dfrA5 were present in three (3.4%) of the isolates. The results showed that of the ESBL-producing isolates, 85.2% (n = 122), 53.2% (n = 76) and 26.1% (n = 37) were blaTEM, blaSHV and CTX-M harboring isolates, respectively. Conclusions: Our study indicated a high frequency of cotrimoxazole resistance gene in E. coli isolates from Ilam and Tehran (Milad) hospitals, and sul genes had a major role in cotrimoxazole resistance of these isolates.

The Frequency of HBB Mutations Among β-Thalassemia Patients in Hamadan Province, Iran

Abstract β-Thalassemia (β-thal) caused by mutations on the HBB gene is the most common single-gene disorder in the world. In this study, the HBB gene mutation was investigated in Hamadan province, Iran. Forty-one patients referred to a referral hospital were admitted to the study. DNA samples were extracted from peripheral blood. The HBB gene was sequenced in all recruited patients. Eleven mutations and eight polymorphisms were found in the studied patients. IVS-II-1 (G>A) (HBB: c.315+1 G>A) was the most common mutation, accounting for 25.61% of mutant alleles. Other mutations included codon 8 (–AA) (HBB: c.25−26delAA); IVS-I-110 (G>A) (HBB: c.93−21 G>A); codons 8/9 (+G) (HBB: c.27−28insG); IVS-I-1 (G>A) (HBB: c.92 G>A); codon 44 (–C) (HBB: c.135delC); codons 25/26 (+T) (HBB: c.78−79insT); IVS-I-130 (G>C) (HBB: c.93−1 G>C); –28 (A>C) (HBB: c.−78 A>C); codons 36/37 (–T) (HBB: c.112delT) and IVS-I-6 (T>C) (HBB: c.92+6 T>C). According to our findings, the IVS-II-1 mutation has the highest prevalence in Hamadan Province. It was found that the total frequency of the IVS-II-1, codons 25/26 (+T), codons 8/9 (+G), IVS-I-110 and IVS-I-1 mutations was 82.92%. Therefore, given these findings, it is recommended that these five mutations are screened for as a first step in laboratories without sequencing instruments, and that the rest of the gene is subsequently examined.

Antimicrobial effect of Pistacia atlantica leaf extract

The antimicrobial effect of the mastic tree (Pistacia atlantica) under in vitro conditions has been reported. Therefore, it is of interest to evaluate the effect of the plant leaf extract (aqueous) on bacterial load in mouth and saliva. The leaf of the Pistacia atlantica plant was collected and cleaned, dried at 40⁰c and then powdered. The extraction was carried out using the maceration method in vacuum with the rotary evaporator device. Bacterial inhibition (Streptococcus species) by the leaf extract was studied using the disc diffusion and embedding sink diffusion methods. The values of MIC and MBC were determined. The collected data was further analyzed using t-test and repeated measure statistical tests. The disc diffusion technique showed a significant inhibitory effect for Pistacia atlantica leaf extract on S. mutans (ATCC 35668) and S. mitis (ATCC 49456) with inhibition zones of 19 and 25 millimeters, respectively. This is for the highest leaf extract concentration used in this study (p<0.01). The values of MIC and MBC for S.mutans was 60, 90 μg/ml and for S. mitis was 75, 110 μg/ml (p<0.01 significance). The leaf extract has no significant effect on S. salivarius (ATCC 13419). Thus, the antimicrobial properties of the aqueous leaf extract from Pistacia atlantica is demonstrated in this study.

Sewage as a rich source of phage study against Pseudomonas aeruginosa PAO

Pseudomonas aeruginosa is a ubiquitous organism which has emerged as a major public health threat in hospital environments. Overuse of antibiotics has significantly exacerbated the emergence of multi-drug resistant bacteria such as P. aeruginosa. Phages are currently being utilized successfully for aquaculture, agriculture and veterinary applications. The aim of this study was to isolate and characterize of lytic P. aeruginosa phage from sewage of Ilam, Iran. Phage was isolated from sewage that was added to the enrichment along with the host and subsequently filtered. Plaque assay was done by using an overlay method (also called the double agar layer method). Purified plaques were then amplified for characterization. Finally, RAPD-PCR method was conducted for genotyping and Transition electron micrograph (TEM) recruited to determine the morphology and phage family. The phage had high concentration and tremendous effects against a variety of clinical and general laboratory strains (ATCC15693) of P. aeruginosa. Among a set of primers in RAPD panel, only P2 and RAPD5 primers, were useful in differentiating the phages. TEM images revealed that the isolated phages were members of the Siphoviridae family. The phage effectiveness and specificity towards target bacteria and potential to control biofilm formations will be investigate in our further studies.

Soluble Flt-1 Gene Delivery in Acute Myeloid Leukemic Cells Mediating a Nonviral Gene Carrier

Vascular endothelial growth factor (VEGF) is a potent angiogenic factor involved in angiogenesis-mediated progression of acute myeloid leukemia (AML). Studies have reported the role of soluble form of fms-like tyrosine kinase (sFlT-1) delivery as an antitumor agent by inhibiting VEGF. This study investigates the outcome of delivery of a VEGF165 antagonist, soluble vascular endothelial growth factor receptor, namely sFLT-1, mediating lipofectamine 2000 in acute myeloid leukemic cells. A recombinant plasmid expressing sFLT-1 was constructed and transfected into the K562 and HL60 cells using lipofectamine 2000 transfection reagent. sFLT-1 expression/secretion in pVAX-sFLT-1 transfected cells was verified by RT-PCR and western blot. MTS assay was carried out to evaluate the effect of sFLT-1 on human umbilical vein endothelial cells and K562 and HL60 cells in vitro. Treatment with pVAX-sFLT-1 showed no association between sFLT-1 and proliferation of infected K562 and HL60 cells, while it demonstrated a significant inhibitory impact on the proliferation of HUVECs. The results of the current study imply that the combination of nonviral gene carrier and sFLT-1 possesses the potential to provide efficient tool for the antiangiogenic gene therapy of AML.

Detection of human coronavirus strain HKU1 in a 2 years old girl with asthma exacerbation caused by acute pharyngitis

Respiratory viral infections can trigger asthma attack which may lead to sever morbidity. In this report, using molecular methods, we show the chronological association between human coronavirus - HKU1 infection and asthma exacerbation in a two years and seven months old asthmatic girl who was not under treatment and was otherwise healthy.

Dynamics of the bacterially expressed conserved immunogenic region of the human respiratory syncytial virus G protein

Despite all efforts, there is still no effective vaccine available against the human respiratory syncytial virus (HRSV) that is a major cause of severe lower respiratory tract disease in infants and the elderly. In this review, we examined the potential of the conserved immunogenic region (residues 122–226) of the HRSV glycoprotein G alone as the inducer of neutralizing antibodies against this virus. The Escherichia coli produced recombinant conserved region of G (designated as G domain), which was used for rabbit immunization. Although rabbit is a semipermissive host for HRSV, our result showed that the polyclonal antibodies against the G domain protein could strongly neutralize the virus (69.3%), suggesting that the G immunogenic region of HRSV alone has a great potential in vaccine development. To our knowledge, this is the first report in which neutralizing antibodies to respiratory syncytial virus have been evoked using bacterially expressed G immunogenic domain protein without any adjuvant.