Adenosine Deaminase 1 as a Biomarker for Diagnosis and Monitoring of Patients with Acute Lymphoblastic Leukemia

Background: Acute lymphoblastic leukemia (ALL) is known as the most prevalent pediatric malignancy all around the world. Identification of specific biomarker is necessary for early diagnosis and effective therapy. It is believed that Adenosine deaminase (ADA) as an enzyme involved in the purine salvage pathway increases in ALL patients. Herein, the quantity and pattern of ADA isoenzymes were surveyed among ALL patients in comparison to healthy subjects. Methods: Serum and RBC samples of three different groups of ALL patients, including newly diagnosed cases without any drugs administration, subjects with the relapsed disease, patients in the remission stage after therapy, and the healthy subjects were enrolled in the study. Then, the activity and pattern of ADA1, ADA2 and ADA1+cp were determined using ADA kit and electrophoresis on SDS-PAGE, respectively. To confirm the presence of ADA enzyme, the fresh serums, extractions from erythrocytes, JM cell line as a human T lymphocyte line and J774 A.1 as mouse monocyte line were electrophoresed on 1.2 agarose gel and stained with the specific dye. Results: The activities of ADA1 isoenzyme and total ADA in new cases and subjects with the relapsed disease were significantly higher than their activities in the patients in the remission stage and healthy controls (p<0.001). The unbounded ADA1 isoenzyme was found to exist in the erythrocyte, lymphocyte and monocyte. But in serum, all the ADA1 was bounded to the cp protein. Conclusions: ADA1 is the key isoenzyme elevating in ALL patients, therefore this isoenzyme could be a useful biomarker to diagnose ALL patients and monitor their therapies. © 2017 Mina Ebrahimi-Rad et al

Adenosine Deaminase 1 as a Biomarker for Diagnosis and Monitoring of Patients with Acute Lymphoblastic Leukemia

Background: Acute lymphoblastic leukemia (ALL) is known as the most prevalent pediatric malignancy all around the world. Identification of specific biomarker is necessary for early diagnosis and effective therapy. It is believed that Adenosine deaminase (ADA) as an enzyme involved in the purine salvage pathway increases in ALL patients. Herein, the quantity and pattern of ADA isoenzymes were surveyed among ALL patients in comparison to healthy subjects. Methods: Serum and RBC samples of three different groups of ALL patients, including newly diagnosed cases without any drugs administration, subjects with the relapsed disease, patients in the remission stage after therapy, and the healthy subjects were enrolled in the study. Then, the activity and pattern of ADA1, ADA2 and ADA1+cp were determined using ADA kit and electrophoresis on SDS-PAGE, respectively. To confirm the presence of ADA enzyme, the fresh serums, extractions from erythrocytes, JM cell line as a human T lymphocyte line and J774 A.1 as mouse monocyte line were electrophoresed on 1.2 agarose gel and stained with the specific dye. Results: The activities of ADA1 isoenzyme and total ADA in new cases and subjects with the relapsed disease were significantly higher than their activities in the patients in the remission stage and healthy controls (p<0.001). The unbounded ADA1 isoenzyme was found to exist in the erythrocyte, lymphocyte and monocyte. But in serum, all the ADA1 was bounded to the cp protein. Conclusions: ADA1 is the key isoenzyme elevating in ALL patients, therefore this isoenzyme could be a useful biomarker to diagnose ALL patients and monitor their therapies. © 2017 Mina Ebrahimi-Rad et al

Evaluation of serum adenosine deaminase and its isoenzymes in patients with ovarian cancer

Ovarian cancer is the most lethal gynecological cancer worldwide. There are great relationships between the activities of adenosine deaminase (ADA), one of the enzymes in purine nucleotide pathway and carcinogenic process. In the present study the activities of the total ADA, ADA1 and ADA2 were measured in the sera of the patients with ovarian cancer. In this study, activities of tADA, ADA1 and ADA2 were assessed in sera of 30 patients with ovarian cancer and 30 normal control individuals, using a modified Ellis method in which only ADA2 activity was measured in the present of a specific inhibitor, erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA). Our results showed that the tADA, ADA1, and ADA2 serum activities of patients were found to be significantly increased (P < 0.05) than those of healthy control group. Although, ADA and its isoenzymes were not the specific markers for diagnosis of ovarian cancer, measurement of their activities may be used as a diagnostic means in ovarian cancer as well as the other analytical procedures

Phenotypic and Genotypic Characterization of ESBL-, AmpC-, and Carbapenemase-Producing Klebsiella pneumoniae and Escherichia coli Isolates

Objectives: Drug resistance among gram-negative bacteria is a worldwide challenge. Due to the importance of drug-resistant Klebsiella pneumoniae and Escherichia coli strains in hospital-acquired infections, we aimed to determine the phenotypic and genotypic characteristics of ESBL-, AmpC-, and carbapenemase-producing isolates obtained from hospitalized patients in Tehran and Ilam (Iran). Materials and Methods: In total, 90 K. pneumoniae isolates and 65 E. coli isolates were collected from various infections. Phenotypic identification of bacterial isolates was performed using standard methods. Phenotypic screening of ESBL, AmpC, and carbapenemase enzymes was carried out. Detection of ESBL, AmpC, and carbapenemase genes was also performed by the PCR method. Results: Phenotypic detection tests showed that 36 (40) K. pneumoniae and 23 (35.4) E. coli isolates were ESBL producers. Moreover, 18 (20) and 6 (9.2) K. pneumoniae and E. coli isolates were AmpC producers, respectively. Modified Hodge test results indicated that 39 (43.3) K. pneumoniae and 18 (27.7) E. coli isolates produced carbapenemase. Molecular tests showed that 40 of K. pneumoniae and 36.9 of E. coli isolates were ESBL positive. AmpC was detected in 24.4 and 13.8 of K. pneumoniae and E. coli isolates. Carbapenemase was detected in 34 (37.8) K. pneumoniae and 13 (20) E. coli isolates. -Conclusion: In this study, 3 K. pneumoniae isolates simultaneously carried ESBL, AmpC, and carbapenemase genes. Up-to-date strategies such as combination therapy or utilization of new antimicrobial agents might help to combat such drug-resistant organisms. © 2019 The Author(s) Published by S. Karger AG, Basel

The rationale behind antibiotic resistance pattern in Klebsiella pneumoniae

Presently, there is an increase in antibiotic resistance in bacteria, due to relax prescription of antibiotics, especially in Iran. Undoubtedly, in toxin antitoxin (TA) system, a toxin neutralized by antitoxin, which known as a potent antimicrobial target; but there is no extensive survey on the prevalence of TA loci in large scale of Klebsiella pneumoniae. Therefore, this study aims to determine the prevalence of different TA loci in clinical and environmental K. pneumoniae isolates. For this reason, 48 K. pneumoniae clinical isolates and 49 K. pneumoniae environmental isolates were subjected for evaluation of different TA loci. The results of current study indicated that there is no association between antibiotic resistances and presence of TA loci in clinical and environmental K. pneumoniae. The role of TA loci as a potent target in antibiotic resistant K. pneumoniae has been complicated. Therefore, more studies should be performed to explain why TA loci are presented in K. pneumoniae and what is the rationale behind antibiotic resistant K. pneumoniae

Genomic Diversity and Virulence Genes among Clinical Isolates of Pseudomonas aeruginosa

Background: Typing of nosocomial pathogens is necessary to determine the source of an outbreak. The aim was to determine the genomic variability among Pseudomonas aeruginosa (P. aeruginosa) by random amplification of polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus (ERIC) methods. Methods: Fifty P. aeruginosa isolates were obtained from the hospitals. The source of these isolates were burn wound and urinary tract infections. After detection of P. aeruginosa by biochemical methods, chromosomal deoxyribonucleic acid (DNA) was extracted by a DNA extraction kit. ERIC-PCR and RAPD- PCR was done by standard methods. The polymerase chain reaction (PCR) products were run and visualized in 1.5 agarose gels stained with ethidium bromide. Results: Fifty P. aeruginosa isolates were analyzed by ERIC-PCR and RAPD-PCR methods. Multiple PCR fragment sizes generated by two PCR methods and PCR product size were between 200 - 3500 bp, and 10 and 7 different PCR patterns were detected by ERIC-PCR and RAPD-PCR, respectively. Eleven isolates were not detected by ERIC-PCR method. Fifteen isolates were typed to a single genotype by the RAPD-PCR method. Conclusions: We suggested that ERIC and RAPD PCR are equally suitable, inexpensive, fast, reproducible, and discriminatory as rapid DNA typing tools for effective epidemiological surveillance of P. aeruginosa isolates. Our results suggest that these DNA typing tools could be used in routine epidemiological surveillance, outbreak surveillance, and in the identification of the source of transmission of P. aeruginosa

Molecular analysis of uropathogenic E.coli isolates from Urinary tract infections in Ilam

The aim of the current study was to investigate the prevalence virulence genes profile in UPEC isolates in Ilam. For this purpose, a total of 8o UPEC isolates for patients with UTIs were collected during 6 months period. The multiplex polymerase chain reaction (Multiplex PCR) was used for detection of the papEF, fimH, iucD, hlyA, fyuA, and ompT genes. The prevalence of genes the fimH, papEF, iucD, fyuA, hlyA, hlyA and ompT were 87.5, 47.5, 60, 67.5, 27.5, 47.5 and 71.2, respectively. Among all of isolates, 27 gene profiles were obtained. The number isolates related to 4 virulence factor simultaneous that were 25 isolates. Considering the different antibiotic patterns, will be proposed develop and an appropriate program for antibiotics in each region and even in each hospital. Also, investigation about the profile of different virulence genes which capable of inducing various pathogens is will be done in the future

Molecular characterization of AmpC β-lactamases among Klebsiella pneumoniae isolated from Ilam and Tehran hospitals, from Iran

The common use of beta lactam antibiotics for tratment of bacteria infections leads to increase the world wide microbial resistance by producing beta lactamase enzyme among clinical isolates. In recent years, the production of broad-spectrum beta-lactamase enzymes in clinical isolates, especially E.coli and Klebsiella bacteria are common. Typical Ampc enzymes (class C- ESBLs) confirm resistance to most oxyimino cephalosporins. The aim of this study is to determine the prevalence of AmpC type extended spectrum beta lactamases genes in clinical isolates of Klebsiella pneumoniae. 108 clinical sample of K.pneumoniae, isolated from hospitalized patients procured from two hospitals in Ilam and Tehran. To identify Ampc genes, PCR method was used. 95/3 percent of isolates were resistant to Cefoxitin, 49 isolates were positive for FOXM cluster genes, 35 were positive for DHAM cluster genes and 6 were positive for CITM cluster genes. Our results showed that among of clinical isolates of Klebsiella pneumoniae, prevalence broad-spectrum beta-lactamase enzymes and Ampc genes are relatively high

Usnic acid improves memory impairment after cerebral ischemia/ reperfusion injuries by anti-neuroinflammatory, anti-oxidant, and anti-apoptotic properties

Objective(s): Cerebral ischemia/reperfusion causes complex pathological mechanisms that lead to brain tissue damage. Usnic acid is a lichen secondary metabolite that has many different biological properties including anti-inflammatory and anti-oxidant activities. Therefore, the objective of the current study was to investigate the neuroprotective effects of usnic acid on apoptotic cell death, neuroinflammation, anti-oxidant enzyme activities, and oxidative stress levels after transient cerebral ischemia/reperfusion. Materials and Methods: Forty-two male Wistar rats were randomly assigned to three groups (sham, ischemia/reperfusion, and ischemia/reperfusion+usnic acid). Ischemia was induced by 20 min occlusion of common carotid arteries. Injection of usnic acid (25 mg/kg, intraperitoneally) and saline was done at the beginning of reperfusion time. Morris water maze was applied to assess spatial memory. The protein expression amount was measured using immunohistochemical and immunofluorescence staining. Spectrophotometric assay was performed to determine the levels of anti-oxidant enzymes. Results: Usnic acid significantly reduced caspase-3, glial fibrillary acidic protein-positive and ionized calcium-binding adaptor molecule 1-positive cells (P<0.001) and enhanced spatial memory disorders (P<0.05) due to brain ischemia. In addition, treatment with usnic acid improves effects in the antioxidant system following cerebral ischemia (P<0.05). Conclusion: Our findings indicate that usnic acid has neuroprotective properties, which possibly is applicable as a promising candidate for cerebral injuries caused by ischemia. © 2020 Mashhad University of Medical Sciences. All rights reserved