The role of oxidative stress in the pathogenesis of phenylketonuria (PKU)-associated disorders has been implicated. Ischemia modified albumin (IMA) is a modified form of serum albumin, which is produced under the conditions of oxidative stress. The aim of this study was to measure the serum level of IMA in the PKU patients and to investigate its ability in predicting the status of oxidative stress in these patients. Fifty treated-PKU patients and fifty age- and sex-matched healthy subjects were included in the study. The blood samples were obtained and the serum level of phenylalanine (Phe) was measured using reverse phase HPLC method. The levels of IMA, malondialdehyde (MDA), gamma-glutamyl transferase (GGT) activity, and uric acid (UA) were determined using colorimetric methods. The levels of serum Phe, IMA, and MDA were significantly higher (p < 0.001) and the level of UA (p < 0.05) was lower in the PKU patients compared to control group. Serum IMA level was positively correlated with MDA (r = 0.585, p < 0.001) and UA (r = 0.6, p < 0.001). An inverse relationship was observed between the serum level of IMA and Phe (r = - 0.410, p < 0. 01). Results of the present study suggest that serum IMA level could be used as a novel marker for the evaluation of oxidative stress in the PKU patients.
Background: Pseudomonas aeruginosa is a Gram negative ubiquitous opportunistic organism and one of the more problematic drug-resistant pathogens encountered today. Objectives: The aims of the current multicenter research were to assess antibiotic resistance profiles, carbapenemase production, and detection of antibiotic resistance IMP gene as well as virulence factors genes including exoA, algD, lasB, and plcH among the clinical isolates of P. aeruginosa. Methods: A total of 80 nonduplicate isolates of P. aeruginosa were recovered from inpatients. Bacterial identification was done by standard diagnostic tests. Species was confirmed by detection of the exoA gene using the PCR technique. Antimicrobial susceptibility test was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Carbapenemase production among the isolates was determined by the modified Hodge test. Virulence genes were detected by PCR. Results: A total of 42 (52.5%) isolates recovered from wound specimens. Colistin was the most effective antibiotic against isolates (97.5% isolates were susceptible) and levofloxacin was the least effective drug (67.5% isolates were resistant). The most common antibiotic resistance pattern was CIPR-CPMR-GEMR (47 isolates). In total, 47 (58.75%) isolates were identified as multidrug resistance (MDR), while 30% of isolates were carbapenemase producer (MHT+). Among studied isolates, plcH+ and lasB+ genotypes (100% isolates) were the most common virulence gene patterns. Of 80 P. aeruginosa isolates, 39 (48.75%) showed algD+, plcH+, lasB+, and IMP+ genotype. The blaIMP resistance gene was detected in all MHT positive and MDR isolates. Conclusions: In our study, the emergence of potentially highly pathogenic and carbapenem-resistant strains in joining with a MDR phenotype is alarming, as a feasible outcome would be a severe clinical result concomitant with critical restrictions in antibiotic therapy.
Background:The p28 is a small-sized cell-penetrating peptide derived from bacterial protein azurin and can function as a cancerspecific anti-proliferative agent. It can penetrate cancer tissues easily without involving the immune system, and increase the intracellular concentration of p53. Objectives: In this study, we have expressed and purified recombinant p28, then evaluated its anti-proliferative and pro-apoptotic effects on Raji cancer cell line. Methods: The p28 gene was amplified and cloned into pTZ57R cloning vector and was sequenced subsequently. Afterward, it was transformed into E. coli BL21 bacterial host by using pET-28a expression vector. Peptide purification was carried out using Ni-NTA chromatography system. Bradford, SDS-PAGE, and western blotting assays were applied to assess the concentration and expression level of the recombinant peptide. The proficiency of p28 in inhibition of tumor growth and induction of apoptosis in cancerous cells was investigated by evaluating the Raji and HEK-293 cells treated with different concentrations of p28. Results: The overexpression of the p28 peptide in the bacterial host was confirmed by SDS-PAGE and western blotting. Moreover, Bradford assay revealed desirable concentrations of the recombinant p28 before and after dialysis. The MTT and PE-Annexin V apoptosis assays indicated the specific function of p28 in impeding the proliferation of cancerous cells and triggering the apoptosis. Conclusions: The p28 induces apoptosis in cancerous cells but not in normal control cells. In summary, p28 is a non-immunogenic small peptide that can penetrate cancerous cells preferentially, impede the cell proliferation, and induce the apoptosis. Overall, these findings suggest p28 as a promising anticancer drug.
Prostate cancer (PCa) is the most frequently diagnosed cancer and the second most common cause of cancer related mortality in United States male population. ScFv fragments have different usefulness. For example they have small size, high perfusion rate, high yield of production and are non-immunogenic, thus they can be used for therapeutic purposes. In this project we used a synthetic human ScFv library for isolation of ScFv monoclonal antibodies against prostate specific membrane antigen. For this purpose, after five rounds of cell-panning, and also five rounds of antigen-panning with rPSMA specific anti- PSMA ScFv-phage particles were isolated. Phages with high affinity toward PSMA were selected and used for further analysis. Specificity and affinity of both ScFv to PSMA and LnCaP cell line examined by ELISA. Recombinant ScFv antibody isolated from cell-panning had higher specificity and affinity for both the antigen and LNCaP cell line. Our result demonstrated that ScFv antibody obtained by cell-panning can target PSMA antigen and cell lines.
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