New Biomarkers in Early Diagnosis of Acute Kidney Injury in Children

en 36504568 New Biomarkers in Early Diagnosis of Acute Kidney Injury in Children Acute Kidney Injury (AKI) is a common condition with a high risk of mortality and morbidity, so, early diagnosis and management of AKI is very important in clinical practice. Despite significant progress in the management of AKI, it still carries high morbidity and mortality. BUN and serum creatinine are not very sensitive nor specific for the diagnosis of AKI because they are affected by many renal and non-renal factors that are independent of kidney injury or kidney function and change significantly only after significant kidney injury and with a substantial time delay. Detection of biomarkers of AKI made predominantly by the injured kidney tissue are essential for the early diagnosis of AKI. An ideal biomarker should be one that could be easily measured, with no interference with other biologic variables, and be able to clarify early phases of kidney damage. The most common biomarkers studied are Neutrophil Gelatinase-Associated Lipocalin (NGAL), Interleukin-18 (IL-18), Kidney Injury Molecule-1 (KIM-1), Cystatin-C, L type Fatty Acid-Binding Protein (L-FABP), N-Acetyl- β-D Glucosaminidase (NAG), netrin-1, vanin-1, and Monocyte Chemoattractant Protein-1 (MCP-1) and calprotectin. Acute kidney injury, Biomarker, Calprotectin, Cystatin C, Interleukin-18, KIM-1, NGAL 264 269 https://www.ajmb.org/En/Article.aspx?id=60513 https://www.ajmb.org/PDF/En/FullText/60513.pdf BehnazBazargani Pediatric Chronic Kidney Disease Research Center, Department of Pediatric Nephrology, Children Medical Center Hospital, Tehran University of Medical Sciences, Tehran, Iran91908MastanehMoghtaderi 91909

en 36504571 Antibodies Produced Toward Recombinant RBD and Nucleocapsid Neutralize SARS-COV-2 Background: The highly contagious SARS-COV-2 virus spread rapidly from China and formed a global pandemic. The virus has infected over 509 million people worldwide and killed about 6.32 million up to date. Up on invasion, the Receptor Binding Domain (RBD) of Spike protein plays a crucial role in the entry of the virus into the host cell. The virus N protein is another protein that has a critical role for genome packaging. Methods: As bioinformatics approaches, the cassette design, codon adaptation, and protein stability were investigated in this study. Synthetic genes of RBD and N were cloned separately in pET28a + expression vector. They were transferred into Escherichia coli (E. coli) BL21 DE3 host cell, and expression of recombinant proteins was induced with IPTG. The recombinant proteins were purified by column chromatography and approved by Western blotting. Animal immunization was performed with each of the recombinant proteins individually and in combination of the two. The antibody titer of the blood serum from control and immunized mice groups was determined by ELISA technique. Finally, the anti-spike neutralization test was performed. Results: The expression and purification of RBD protein were monitored on SDS-PAGE, two bands of about 28 and 45 kDa for RBD and N appeared on gel distinctly, which were further validated by Western blotting. According to ELISA results, related antibodies were traced to a dilution of 1/64000 in immunized sera. The neutralization test exhibited produced antibodies' potency to bind the virus proteins. Using SPSS software, statistical analysis was performed by Duncan's test and T-test. Conclusion: According to the present study, recombinant proteins, either RBD alone or in combination with N adequately stimulated the immune response, and the raised antibodies could neutralize the virus in in vitro test. Coronavirus, Nucleocapsid, Recombinant vaccines, SARS-CoV-2, Spike glycoprotein 270 277 https://www.ajmb.org/En/Article.aspx?id=60514 https://www.ajmb.org/PDF/En/FullText/60514.pdf AmirRezaeiDepartment of Biology, Shahed University, Tehran, Iran91910ShahramNazarian11429HosseinSamiei AbianehDepartment of Biology, Imam Hussein University, Tehran, Iran91912EmadKordbachehDepartment of Biology, Imam Hussein University, Tehran, Iran91903ZahraAlizadehDepartment of Biology, Shahed University, Tehran, Iran91914Seyed LatifMousavi Gargari91915

en 36504565 A Panel of Circulating microRNAs as a Potential Biomarker for the Early Detection of Gastric Cancer Background: The high mortality rate of Gastric Cancer (GC) is a consequence of delayed diagnosis. The early diagnosis of GC could increase the five-year survival rate among patients. We aimed to find a panel of microRNAs (miRNA) for the detection of GC in the early stages. Methods: In this case-control study, we selected consistently upregulated miRNAs from the results of 12 high-throughput miRNA profiling studies in GC. In the profiling phase, the differential expressions of 13 candidate miRNAs were analyzed by quantitative reverse-transcription PCR (qRT-PCR) in two pooled RNA samples prepared from the plasma of eight GC patients and eight matched controls. In the validation phase, significantly upregulated miRNAs from the profiling phase were further evaluated in the plasma samples of 97 patients with stage I-IV gastric adenocarcinoma and 100 healthy controls. Results: In the profiling phase, six miRNAs (miR-18a, 21, 25, 92a, 125b and 221) were significantly upregulated in the GC patients compared to the controls (p<0.05). However, in the validation phase, only significant up-regulation of miR-18a, 21 and 125b was confirmed (p<0.05). A panel of miR-18a/21/125b was able to detect GC patients with stage I-IV from the controls (p<0.001; AUC=0.92, sensitivity=86%; specificity=85%). In addition, the panel could distinguish the early-stage GC (I+II) from the control group with an AUC of 0.83, a sensitivity of 83%, and a specificity of 75%. Conclusion: A panel of circulating miR18a/21/125b could be suggested as a potential biomarker for the early detection of GC. Biomarker, Circulating microRNA, Detection, Gastric cancer, Gene expression 278 286 https://www.ajmb.org/En/Article.aspx?id=60515 https://www.ajmb.org/PDF/En/FullText/60515.pdf KioomarsSaliminejadReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran393HabibollahMahmoodzadehDepartment of Surgery, Cancer Institute, Imam Khomeini Hospital, Tehran University of Medical Sciences, Tehran, Iran31506ShahrzadSoleymani FardHematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran92MarjanYaghmaeiHematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran373Hamid RezaKhorram KhorshidGenetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran42Seyed AsadollahMousaviHematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran31507MohammadVaezi Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran91920Seyed HamidollahGhaffari31570

en 36504564 Bioactive Materials Derived from Menstrual Blood Stem Cells Enhance the Quality of In Vitro Bovine Embryos Backgrounds: The aim of this study was to determine whether the addition of bioactive materials derived from Menstrual Blood Stem Cells (MenSCs) to the oocyte maturation medium may improve the quality of bovine embryos in vitro. Methods: MenSCs were collected from 6 healthy women (between 26 and 36 years old) and after 3 days of culture, their bioactive materials were frozen. The bovine Cumulus-Oocyte-Complexes (COCs) were aspirated from ovarian slaughterhouse and the oocytes with more than three layers of cumulus cells were cultured in vitro in media supplemented with (treatment) and without (control) 10% MenSCs’ bioactive materials. After IVM/IVF, the presumptive zygotes were cultured for 8 days. Results: The blastocyst rate on day 8 in treatment group was higher than control (40.2±1.9 vs. 23±4.2.3, p=0.001). The ratio of Trophectoderm (TE) and Inner Cell Mass (ICM) (ICM/TE) cells was also greater in treatment group compared to control (30.3±2 vs. 14.9±1; p=0.001). The re-expansion of vitrified blastocysts, 24 hours after warming, in treatment group was higher than control (93.3±2.5 vs. 66.2±8.8; p=0.01). The expression of some genes related to pluripotency and implantation (OCT4, CDX2, and IFNT) were increased in treatment group compared to control (p<0/05). Conclusion: In conclusion, the addition of MenSCs’ bioactive materials during in vitro maturation of bovine oocytes could improve the quantity and quality of bovine IVP embryos. Also, the expression of some genes associated with pluripotency and implantation in the blastocyst would be increased. Bovine, Embryo, In vitro production, IVM, Menstrual blood stem cells 287 293 https://www.ajmb.org/En/Article.aspx?id=60516 https://www.ajmb.org/PDF/En/FullText/60516.pdf Mohammad SobhanAmini Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran91922Mohammad MehdiNaderi334AbolfazlShiraziReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran82MehdiAminafsharDepartment of Animal Science, Science and Research Branch, Islamic Azad University, Tehran, Iran91923SaraBorjian BoroujeniReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran337MostafaPournourali Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran91924AliMalekpour Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran91925

en 36504569 Evaluation of PLGA-Encapsulated Recombinant GroEL of S. typhi immune Responses Against Enterohaemorrhagic and Enteropathogenic Escherichia coli Background: Heat Shock Proteins (HSPs) elicit humoral and cellular immune responses. Due to their high sequence homology, they can be developed as a new immunogen for cross prophylactic and vaccination effects against infectious agents such as Enteropathogenic and Enterohemorrhagic Escherichia coli (EPEC and EHEC). This study aimed to evaluate the immunogenicity and cross-protective efficacy of rGroEL of Salmonella typhi (S. typhi) encapsulated in poly lactic-co-glycolic acid (PLGA) nanoparticles against EPEC and EHEC. Methods: Recombinant GroEL was expressed in Escherichia coli (E. coli) and purified using Ni-NTA affinity chromatography. The protein was encapsulated in PLGA by the double emulsion method, and the nanoparticles were characterized physicochemically. BALB/c mice were immunized, and the efficacy of the protein to elicit immune responses was assessed. Results: Over-expression in E. coli led to corresponding 64.5 kDa protein bands in Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Non-ag-gregated nanoparticles had a spherical shape with a mean diameter of 194.3±3 nm and encapsulation efficiency of 89.5±2.5%. Antibody isotyping revealed that GroEL immunization induced both IgG1 and IgG2a antibodies. Moreover, immunization of the mice with recombinant GroEL protein conferred 80 and 60% protection against lethal infections by EPEC and EHEC, respectively. Furthermore, organ burden studies revealed a significant reduction in infection in the immunized mice compared to the non-immunized ones. Passive immunization with anti-GroEL sera also protected 50% of the mice against the lethal doses of EHEC and EPEC strains. Conclusion: The findings indicated that immunization of the mice with recombinant GroEL of S. typhi elicited cross-protection against other bacterial infections. This represented the immense potential of GroEL to be developed as a single vaccine against multiple pathogens Heat-shock proteins, Immunogenicity, Nanoparticles, Salmonella typhi, Vaccines 294 302 https://www.ajmb.org/En/Article.aspx?id=60517 https://www.ajmb.org/PDF/En/FullText/60517.pdf MiladParvane Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran91926ShahramNazarian11429EmadKordbachehBiology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran91903JavadFathiStudent Research Committee, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran91902Mohamad EbrahimMinae 91929Mohammad RezaRamezani Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran91930

en 36504570 Comparative Antioxidant and Anti-gout Activities of Citrullus colocynthis loaded Fruit Silver nanoparticles with its Ethanolic extract Background: The biological synthesis of silver nanoparticles (AgNPs) using plant materials is a rapidly developing method with several alternative medical applications. This comparative study of ethanolic fruit extract of Citrullus colocynthis (C. colocynthis) (EFECC) and synthesized silver nanoparticles (CC-AgNPs) were carried out for antioxidants and anti-gout arthritic activities. Methods: The AgNPs were synthesized using C. colocynthis fruit and its characterization was done by UV-visible spectroscopy, TEM, XRD and FT-IR. The 90% ethanol was used for extract preparation. Antioxidant activity was analyzed by DPPH and the Hydrogen Peroxide (H2O2) method. In vitro anti-arthritic activity was tested by xanthine oxidase inhibition, protein denaturation and HRBC membrane stabilization assay. Results: The synthesized CC-AgNPs were confirmed by UV-vis spectroscopy and TEM images displayed spherical shapes with 10-45 nm size range. Furthermore, the functional groups and crystalline structure of CC-AgNPs were determined by FT-IR and XRD analysis. The biosynthesized CC-AgNPs exhibited an excellent free radical scavenging ability than EFECC. In anti-arthritic activity, the CC-AgNPs showed effective inhibition of xanthine oxidase production, protein denaturation, and damaged RBC membranes compared to EFECC. Conclusion: The antioxidant activities and in vitro anti-arthritic assays revealed that CC-AgNPs are better anti-gout agents than EFECC. This research suggested that biosynthesized silver nanoparticles from C. colocynthis fruit are an important target in the field of anti-gout drug discovery. Citrullus colocynthis, Metal nanoparticles, Silver, Spectrophotometry, Ultraviolet, Xanthine oxidase 303 309 https://www.ajmb.org/En/Article.aspx?id=60518 https://www.ajmb.org/PDF/En/FullText/60518.pdf SuganyaKarunakaranDepartment of Biotechnology, Dr. MGR Educational & Research Institute, deemed to be University, Maduravoyal, Chennai, India91946RajeswaryHari91947

en 36504563 Association between PTCH1 and RAD54B Single-Nucleotide Polymorphisms and Non-syndromic Orofacial Clefts in the Northeast Population of Iran Background: Non-Syndromic Cleft Lip with or without cleft Palate (NSCL/P) is a common developmental disorder of the head and neck with a multifactorial etiology. The current study aimed to evaluate the potential association of PTCH1 (rs10512248) and RAD54B (rs12681366) polymorphisms with NSCL/P in the Northeast Iranian population. Methods: In the present study, blood samples were taken from 122 subjects with NSCL/P and 161 healthy controls. Polymerase Chain Reaction (PCR) followed by Restriction Fragment Length Polymorphism (RFLP) were used to conduct genotyping of single-nucleotide polymorphisms. Results: Although differences were observed between cases and controls in rs10512248 and rs12681366, our data did not support a significant association of these polymorphisms with NSCL/P in our population. Conclusion: Our findings suggest that polymorphisms of rs10512248 and rs12681366 may not be potential risk factors for NSCL/P in the Northeast Iranian population due to the multifactorial and multiethnicity characteristics of some genes. Cleft lip, Cleft palate, Polymorphism, PTCH1, RAD54B 310 316 https://www.ajmb.org/En/Article.aspx?id=60519 https://www.ajmb.org/PDF/En/FullText/60519.pdf RezaMorvaridi Farimani Department of Orthodontics, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran91939MohsenAzimi-Nezhad UMR, INSERM U 1122, IGE-PCV, Interaction Géne-Environment Enpathophysiologie Cardiovasculaire, Université De Lorraine, Nancy, France91940Hamid RezaKhorram KhorshidGenetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran42AsgharEbadifar Dentofacial Deformities Research Center Research Institute of Dental Sciences, Faculty of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran91941SabaTohidkhah Tehran University of Medical Sciences, Tehran, Iran91942ZahraJafarian Iranian Research Center on Aging, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran91943KooroshKamaliDepartment of Public Health, Faculty of Public Health, Zanjan University of Medical Sciences, Zanjan, Iran89ZeinabNazari Non-Communicable Diseases Research Center, Neyshabur University of Medical Sciences, Neyshabur, Iran91944RezaEbrahimzadeh-Vesal Pardis Genetic Laboratory, Tehran, Iran91945

en 36504567 Effects of Poly-N-isopropylacrylamide Microgels Containing Antibiofilm Substances on Pseudomonas aeruginosa Isolated from Chronic Wounds Background: Biofilm formation helps Pseudomonas &lrm;aeruginosa (P. &lrm;aeruginosa) survive in various environments. Microgels can be effective in treatment of bacterial infections. The major aim of this study was to investigate effects of poly-N-isopropyl-acrylamide microgels (PNIPAM) on P. aeruginosa. Methods: Totally&lrm;, &lrm;100 P. aeruginosa strains were isolated from chronic wound infections&lrm;. Quantitative assessments of biofilm formation and antibiotic susceptibility were carried out. Furthermore, algD, lasR, and PA2714 genes were amplified to investigate gene frequencies and expression rates. Results: Significant decreases were seen in lasR expression in EDTA-treated samples&lrm;. Significant decreases were observed in expression of algD and lasR treated with xylitol. Decreased &lrm;expression of PA2714 was seen in samples treated with xylitol with no significance.  Conclusion: The PNIPAM containing xylitol &lrm;or EDTA could penetrate biofilms of P. aeruginosa and significantly decrease expression of lasR and algD. This can be a novel strategy in the management of chronic wounds. Biofilms, Microgels, Pseudomonas aeruginosa, Real-time PCR 317 320 https://www.ajmb.org/En/Article.aspx?id=60520 https://www.ajmb.org/PDF/En/FullText/60520.pdf AkramEtemadinia Department of Pathobiology, School of Public Health, Tehran University of Medical Science, Tehran, Iran91934AmirSeyfoori Department of Biomaterials and Tissue Engineering, Breast Cancer Research Center, Motamed Cancer Institute, ACECR, Tehran, Iran91935AbbasRahimi Foroushani Department of Pathobiology, School of Public Health, Tehran University of Medical Science, Tehran, Iran91936RaminMazaheri Nezhad FardFood Microbiology Research Center, Tehran University of Medical Science, Tehran, Iran81819RonakBakhtiari 91938

en 36504566 Mitochondrial Transfer from Menstrual Blood Stromal/Stem Cells Promotes Survival of Cardiomyocytes Following Myocardial Infarction Dear editor, Recently, Mitochondrial Transfer (MT) from stem cells to injured cells has been proposed as a novel therapeutic strategy that could restore the bioenergetics requirement of damaged tissues 1. This organelle is essential for cellular homeostasis, cell survival, cell growth, cell death induction, calcium storage, cell signaling, and energy supply, especially in energy-demanding cells like cardiomyocytes 2,3. Mitochondrial dysfunction is contributed to a majority of pathologic conditions like Myocardial Infarction (MI) and cardiomyopathies 4. Mitochondrial impairment results in reduction of Adenosine Triphosphate (ATP) production, induces the generation of intracellular Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS), and activates the caspase cleavage pathway 5. It has been confirmed that Mesenchymal Stem Cells (MSCs) could transport mitochondria to a range of cells including endothelial cells and cardiomyocytes 6. It appears that healthy mitochondrial donation by MSCs is a unique phenomenon that leads to replacement of dysfunctional mitochondria in injured cells. It has been designated that the transfer of even a small number of Multipotent Mesenchymal Stem Cells (MMSC)-derived mitochondria resulted in enhanced oxidative phosphorylation and promotion of cell proliferation in the recipient damaged cells 7. Although mitochondrial transmission from various sources of stem cells like Bone Marrow Mesenchymal Stem Cell (BM-MSCs), induced Pluripotent Stem Cells (iPSCs), and Dental Pulp Derived Mesenchymal Stem Cell (DP-MSCs) has been stated, there is no report about MT from menstrual blood Stromal/Stem Cells (MenSCs). Recently we have perceived that, transepicardial MenSCs administration could improve cardiac function, prevent metaplastic development, and promote survival of cardiomyocytes following MI conceivably by transfer of their mitochondria to preserved cardiomyocytes and endothelial cells. We tracked the injected MenSCs 28 days’ post-transplantation by anti-human mitochondrial antibody in MI site in rat model and demonstrated successfully transferred human mitochondria from MenSCs into the targeted cells. Researchers have showed that mitochondrial dysfunction plays critical role in the loss of cardiomyocytes during MI 8. Although exact mechanisms of MT have not been clarified yet, it has indicated that the local microenvironment of injured tissue releases physiological signals that trigger MT 9. For instance, mitochondrial DNA (mtDNA) released by damaged cells is engulfed by MSCs and that later, prompts the cytoprotective function of MSCs and boosts mitochondrial biogenesis 10. Furthermore, ROS and RNS can also stimulate mitochondrial donation 11. It is assumed that, transmission of mitochondria derived from MenSCs may lead to maintenance of cellular homeostasis, preservation of aerobic respiration, reduction the level of ROS, prevention of cell death, and upregulation of cardio-protective cytokines in the cardiomyocytes 12. Mitochondria in MSCs like MenSCs is in dormant state due to lesser energy demands. However upon differentiation, increase in levels of respiratory enzymes, greater oxygen consumption rate, augmented levels of intracellular ATP, increase in mtDNA copy number and mRNA levels may occur 13,14. Interestingly, it has been indicated that the MT phenomenon from MSCs not only results in protection of injured targeted cells, but also, it can lead to more MSCs survival due to decreasing their partially depolarized and dysfunctional mitochondria 15. Researchers indicated that transmission of mitochondria from BM-MSCs to cardiomyocytes can inhibit apoptosis and reprogrammed differentiated cardiomyocytes to progenitor-like cells 16. Furthermore, iPSC-MSCs directly protects cardiomyocytes against induced cardiomyopathy through bioenergetic preservation by functional MT 17. Also MT from MSCs to endothelial cell rescued the injured endothelial cell by reducing apoptosis and promoting proliferation 18. Meanwhile, some resident cells in injured site could transfer mitochondria to injured cells; we believe that only endogenous transfer is transient and cannot inhibit progressive injuries following MI. Our studies have shown that; administration of MenSCs post-MI modifies the metabolism and restores the functionality of heart, and also protect myocardium from further subsequent injuries mainly with donation of their healthy mitochondria to cardiomyocytes and endothelial cells. It is likely that the donor mitochondria fuse with mitochondria in the recipient cell, and restore bioenergetic requirements; or the recipient cell removes its injured mitochondria and gets the donated healthy mitochondria 19. Researchers revealed that human mitochondrial DNA from MSCs could be found in mice 28 days after MSC administration. However, MSC nuclear DNA was not detected 3 days post administration and suggesting that the long-term therapeutic effects of MSCs administration may be related to MT 15. So, the stem cell-based mitochondria transfer approach from MenSCs can be considered as a newly effective therapeutic strategy to treat cardiomyopathies. However, more investigation is needed to explore the exact mechanism of the MenSCs-derived mitochondria communication with the recipient cells, restoration of mitochondrial bioenergetics in these cells, cell-signaling pathways involved to this phenomenon, and understand how this organelle donation would lead to regeneration. Donation, Mitochondria, Myocardial infarction, Stem cells 321 322 https://www.ajmb.org/En/Article.aspx?id=60521 https://www.ajmb.org/PDF/En/FullText/60521.pdf MahmoodManshoriNanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran91931SomaiehKazemnejadNanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran91932HannanehGolshahi 91933