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1.
Weekly Epidemiological Monitor. 2019; 12 (1): 1
in English | IMEMR | ID: emr-199434

ABSTRACT

On 12th December 2018, an expert meet-ing on one health framework for action on emerging zoonotic infections was held in Amman, Jordan. The meeting, orga-nized by the World Health Organization Regional Office for Eastern Mediterrane-an [WHO EMRO], reviewed the frame-work in the light of improving efficient and coordinated multisectoral "one health" approach among human health, animal and environment sectors to pre-vent, detect and respond to emerging zoonotic infection


Subject(s)
Communicable Diseases , One Health , Mediterranean Region
2.
Cell Journal [Yakhteh]. 2019; 20 (4): 450-458
in English | IMEMR | ID: emr-199613

ABSTRACT

Objective: Mesenchymal stem cells [MSC] from various sources have the potentials to positively affect regenerative medicine.Furthermore, pre-conditioning strategies with different agents could improve the efficacy of cell therapy. This study compares the effects of an anti-inflammatory and antioxidant agent, melatonin, on protection of bone marrow-derived MSCs [BMSCs] and adipose tissue-derived MSCs [ADSCs]


Materials and Methods: In this experimental study, rat BMSCs and ADSCs were isolated and expanded. Pre-conditioning was performed with 5 ìM melatonin for 24 hours. Cell proliferation and viability were detected by MTT assay. Expression of BAX, BCL2, melatonin receptors and osteocalcin genes were evaluated by reverse transcriptase-polymerase chain reaction [RT-PCR]. Also, apoptosis was detected with tunnel assay. Osteogenic differentiation was analyzed using alizarin red staining


Results: No significant increase was found in cell viability between BMSCs and ADSCs after melatonin preconditioning. Following melatonin preconditioning, BAX expression was significantly down-regulated in both ADSCs and BMSCs [P<0.05], with the difference being more significant in ADSCs compared to BMSCs. BCL2 expression was increased significantly in both cell types after preconditioning. Metalothionine 1 and Metalothionine 2 were both upregulated significantly in the two cell types [P<0.05]. Melatonin increased osteogenesis capability through increasing osteocalcin expression. However, expression of osteocalcin in BMSCs before and after preconditioning was higher than that in ADSCs. On the other hand, melatonin expression in ADSCs was in higher levels than in BMSCs. Melatonin also improved alizarin red concentration significantly in both BMSCs and ADSCs [P<0.05]. Alizarin red staining severity increased significantly in ADSCs after preconditioning compared to BMSCs [P<0.05]


Conclusion: Here we have shown that the effects of preconditioning on melatonin expression in ADSCs are higher than those in BMSCs. These findings could be used in adoption of a proper preconditioning protocol based on the sources of MSCs in specific clinical applications, especially in bone regeneration

3.
Cell Journal [Yakhteh]. 2019; 20 (4): 459-468
in English | IMEMR | ID: emr-199614

ABSTRACT

Objective: Human amniotic membrane [HAM] is used as a supporter for limbal stem cell [LSC] expansion and corneal surgery. The aim of study is to use HAM extracts from healthy donors to enhance proliferation of LSCs in vitro and in vivo


Materials and Methods: In this interventional experimental study, the effective and cytotoxic doses of the amniotic membrane extract eye drops [AMEED] was assessed by adding different concentrations of AMEED [0-2.0 mg/ml] to LSC cultures for 14 days. Subsequently, the expression levels of ATP-binding cassette sub-family G member 2 [ABCG2, a putative stem cell marker], cytokeratin 3 [K3, corneal maker], K12 and K19 [corneal-conjunctival cell makers] were assessed by real-time polymerase chain reaction [PCR]. In the second step, the corneal epithelium of 10 rabbits was mechanically removed, and the right eye of each rabbit was treated with 1 mg/ml AMEED [every 2 hours [group 1] or every 6 hours [group 2]]. The left eyes only received an antibiotic. The corneal healing process, conjunctival infection, degree of eyelid oedema, degree of photophobia, and discharge scores were evaluated during daily assessments. Finally, corneal tissues were biopsied for pathologic evidences


Results: In comparison to the positive control [10% foetal bovine serum [FBS]], 0.1-1 mg/ml AMEED induced LSC proliferation, upregulated ABCG2, and downregulated K3. There were no remarkable differences in the expression levels of K12 and K19 [P>0.05]. Interestingly, in the rabbits treated with AMEED, the epithelium healing duration decreased from 4 days in the control group to 3 days in the two AMEED groups, with lower mean degrees of eyelid oedema, chemosis, and infection compared to the control group. No pathologic abnormalities were observed in either of the AMEED groups


Conclusion: AMEED increases LSCs proliferation ex vivo and accelerates corneal epithelium healing in vivo without any adverse effects. It could be used as a supplement for LSC expansion in cell therapy

4.
Cell Journal [Yakhteh]. 2019; 20 (4): 469-476
in English | IMEMR | ID: emr-199615

ABSTRACT

Objective: The ability to generate lung alveolar epithelial type II [ATII] cells from pluripotent stem cells [PSCs] enables the study of lung development, regenerative medicine, and modeling of lung diseases. The establishment of defined, scalable differentiation methods is a step toward this goal. This study intends to investigate the competency of small molecule induced mouse embryonic stem cell-derived definitive endoderm [mESC-DE] cells towards ATII cells


Materials and Methods: In this experimental study, we designed a two-step differentiation protocol. mESC line Royan B20 [RB20] was induced to differentiate into DE [6 days] and then into ATII cells [9 days] by using an adherent culture method. To induce differentiation, we treated the mESCs for 6 days in serum-free differentiation [SFD] media and induced them with 200 nM small molecule inducer of definitive endoderm 2 [IDE2]. For days 7-15 [9 days] of induction, we treated the resultant DE cells with new differentiation media comprised of 100 ng/ml fibroblast growth factor [FGF2] [group F], 0.5 micro g/ml hydrocortisone [group H], and A549 conditioned medium [A549 CM] [group CM] in SFD media. Seven different combinations of factors were tested to assess the efficiencies of these factors to promote differentiation. The expressions of DE- and ATII-specific markers were investigated during each differentiation step


Results: Although both F and H [alone and in combination] promoted differentiation through ATII-like cells, the highest percentage of surfactant protein C [SP-C] expressing cells [37%] were produced in DE-like cells treated by F+H+CM. Ultrastructural analyses also confirmed the presence of lamellar bodies [LB] in the ATII-like cells


Conclusion: These results suggest that hydrocortisone can be a promoting factor in alveolar fate differentiation of IDE2- induced mESC-DE cells. These cells have potential for drug screening and cell-replacement therapies

5.
Cell Journal [Yakhteh]. 2019; 20 (4): 477-482
in English | IMEMR | ID: emr-199616

ABSTRACT

Objective: Type 1 diabetes is caused by destruction of beta cells of pancreas. Vildagliptin [VG], a dipeptidyl peptidase IV [DPP IV] inhibitor, is an anti-diabetic drug, which increases beta cell mass. In the present study, the effects of VG on generation of insulin-producing cells [IPCs] from adipose-derived mesenchymal stem cells [ASCs] is investigated


Materials and Methods: In this experimental study, ASCs were isolated and after characterization were exposed to differentiation media with or without VG. The presence of IPCs was confirmed by morphological analysis and gene expression [Pdx-1, Glut-2 and Insulin]. Newport Green staining was used to determine insulin-positive cells. Insulin secretion under different concentrations of glucose was measured using radioimmunoassay method


Results: In the presence of VG the morphology of differentiated cells was similar to the pancreatic islet cells. Expression of Pdx-1, Glut-2 and Insulin genes in VG-treated cells was significantly higher than the cells exposed to induction media only. Insulin release from VG-treated ASCs showed a nearly 3.6 fold [P<0.05] increase when exposed to a highglucose medium in comparison to untreated ASCs. The percentage of insulin-positive cells in the VG-treated cells was approximately 2.9-fold higher than the untreated ASCs


Conclusion: The present study has demonstrated that VG elevates differentiation of ASCs into IPCs. Improvement of this protocol may be used in cell therapy in diabetic patients

6.
Cell Journal [Yakhteh]. 2019; 20 (4): 483-495
in English | IMEMR | ID: emr-199617

ABSTRACT

Objective: Using mesenchymal stem cells [MSCs] is regarded as a new therapeutic approach for improving fibrotic diseases.The aim of this study to evaluate the feasibility and safety of systemic infusion of autologous adipose tissue-derived MSCs [AD-MSCs] in peritoneal dialysis [PD] patients with expected peritoneal fibrosis


Materials and Methods: This study was a prospective, open-label, non-randomized, placebo-free, phase I clinical trial. Case group consisted of nine eligible renal failure patients with more than two years of history of being on PD. Autologous AD-MSCs were obtained through lipoaspiration and expanded under good manufacturing practice conditions. Patients received 1.2 +/- 0.1×106 cell/kg of AD-MSCs via cubital vein and then were followed for six months at time points of baseline, and then 3 weeks, 6 weeks, 12 weeks, 16 weeks and 24 weeks after infusion. Clinical, biochemical and peritoneal equilibration test [PET] were performed to assess the safety and probable change in peritoneal solute transport parameters


Results: No serious adverse events and no catheter-related complications were found in the participants. 14 minor reported adverse events were self-limited or subsided after supportive treatment. One patient developed an episode of peritonitis and another patient experienced exit site infection, which did not appear to be related to the procedure. A significant decrease in the rate of solute transport across peritoneal membrane was detected by PET [D/P cr=0.77 vs. 0.73, P=0.02]


Conclusion: This study, for the first time, showed the feasibility and safety of AD-MSCs in PD patients and the potentials for positive changes in solute transport. Further studies with larger samples, longer follow-up, and randomized blind control groups to elucidate the most effective route, frequency and dose of MSCs administration, are necessary [Registration Number: IRCT2015052415841N2]

7.
Cell Journal [Yakhteh]. 2019; 20 (4): 496-504
in English | IMEMR | ID: emr-199618

ABSTRACT

Objective: Cardiovascular progenitor cells [CPCs] are introduced as one of the promising cell sources for preclinical studies and regenerative medicine. One of the earliest type of CPCs is cardiogenic mesoderm cells [CMCs], which have the capability to generate all types of cardiac lineage derivatives. In order to benefit from CMCs, development of an efficient culture strategy is required. We aim to explore an optimized culture condition that uses human embryonic stem cell [hESC]-derived CMCs


Materials and Methods: In this experimental study, hESCs were expanded and induced toward cardiac lineage in a suspension culture. Mesoderm posterior 1-positive [MESP1+] CMCs were subjected to four different culture conditions: i. Suspension culture of CMC spheroids, ii. Adherent culture of CMC spheroids, iii. Adherent culture of single CMCs using gelatin, and iv. Adherent culture of single CMCs using Matrigel


Results: Although, we observed no substantial changes in the percentage of MESP1+ cells in different culture conditions, there were significantly higher viability and total cell numbers in CMCs cultured on Matrigel [condition iv] compared to the other groups. CMCs cultivated on Matrigel maintained their progenitor cell signature, which included the tendency for cardiogenic differentiation


Conclusion: These results showed the efficacy of an adherent culture on Matrigel for hESC-derived CMCs, which would facilitate their use for future applications

8.
Cell Journal [Yakhteh]. 2019; 20 (4): 505-512
in English | IMEMR | ID: emr-199619

ABSTRACT

Objective: Non-obstructive azoospermia is mostly irreversible. Efforts to cure this type of infertility have led to the application of stem cells in the reproduction field. In the present study, testicular cell-mediated differentiation of male germ-like cells from bone marrow-derived mesenchymal stem cells [BM-MSCs] in an in vitro indirect co-culture system is investigated


Materials and Methods: In this experimental study, mouse BM-MSCs were isolated and cultured up to passage three. Identification of the cells was evaluated using specific surface markers by flow-cytometry technique. Four experimental groups were investigated: control, treatment with retinoic acid [RA], indirect co-culture with testicular cells, and combination of RA and indirect co-culture with testicular cells. Finally, following differentiation, the quantitative expression of germ cell-specific markers including Dazl, Piwil2 and Stra8 were evaluated by real-time polymerase chain reaction [PCR]


Results: Molecular analysis revealed a significant increase in Dazl expression in the indirect co-culture with testicular cells group in comparison to the control group. Quantitative expression level of Piwil2 was not significantly changed in comparison to the control group. Stra8 expression was significantly higher in RA group in comparison to other groups


Conclusion: Indirect co-culture of BM-MSCs in the presence of testicular cells leads to expression of male germ cell-specific gene, Dazl, in the induced cells. Combination of co-culture with testicular cells and RA did not show any positive effect on the specific gene expressions

9.
Cell Journal [Yakhteh]. 2019; 20 (4): 513-520
in English | IMEMR | ID: emr-199620

ABSTRACT

Objective: In vitro transplantation [IVT] of spermatogonial stem cells [SSCs] is one of the most recent methods in transplantation in recent decades. In this study, IVT and SSCs homing on seminiferous tubules of host testis in organ culture have been studied


Materials and Methods: In this experimental study, human SSCs were isolated and their identities were confirmed by tracking their promyelocytic leukemia zinc finger [PLZF] protein. These cells were transplanted to adult azoospermia mouse testes using two methods, namely, IVT and in vivo transplantation as transplantation groups, and testes without transplantation of cells were assigned in the control group. Then histomorphometric, immunohistochemical and molecular studies were done after 2 weeks


Results: After two weeks, histomorphometric studies revealed that the number of subsided spermatogonial cells [SCs] and the percentage of tubules with subsided SCs in IVT and in vivo groups were significantly more than those in the control group [P<0.05]. Immunohistochemical studies in the transplantation groups confirmed that the PLZF protein was expressed in the cells subsided on the seminiferous tubule. Quantitative reverse-transcription polymerase chain reaction [qRT-PCR] demonstrated that the PLZF gene expression was only positive in the transplantation groups, but it was not significantly different between the IVT group and the in vivo group [P>0.05]


Conclusion: Testicular tissue culture conditions after SSC transplantation can help these cells subside on the seminiferous tubule basement membrane

10.
Cell Journal [Yakhteh]. 2019; 20 (4): 521-526
in English | IMEMR | ID: emr-199621

ABSTRACT

Objective: The incidence rate of testicular cancer among young males is high. Co-administration of bleomycin, etoposide and cisplatin [BEP] has increased survival rate of patients with testicular cancer. Although BEP is one of the most effective treatment for testicular cancer, but it severely affects the reproductive system that ultimately leads to infertility. In addition to its antioxidant activity, zinc has an important role in progression of spermiogenesis. This study aimed to evaluate the effect of zinc on sperm parameters, chromatin condensation and testicular structure after BEP treatment


Materials and Methods: In this experimental study, 40 male rats were divided into 4 groups [control, BEP, BEP+ zinc and zinc] and examined for 2 spermatogenesis periods [i.e. 18 weeks]. The rats in BEP and BEP+ zinc group were treated with BEP at appropriate doses [0.75, 7.5, and 1.5 mg/kg] for three cycles of three weeks. Zinc at a dose of 10 mg/kg/day was administered to BEP+ zinc and zinc groups. After 18 weeks, we assessed sperm parameters, and excessive histone in sperm chromatin using aniline blue staining, as well as testicular structure and germ line cells using periodic acid-Schiff staining


Results: After BEP treatment, significant decreases were observed in normal sperm morphology, motility, and concentration, as well as alterations in rat sperm chromatin condensation and testicular tissue [P<0.001]. Furthermore, after zinc consumption for 9 weeks, we observed significant improvements of sperm parameters and chromatin condensation as well as a significant retrieval of spermatogonia, leydig cells and tubular architecture [P<0.05]


Conclusion: Zinc administration after chemotherapy with BEP in testicular cancer might be potentially useful in declining the off target consequence associated with oxidative stress

11.
Cell Journal [Yakhteh]. 2019; 20 (4): 527-536
in English | IMEMR | ID: emr-199622

ABSTRACT

Objective: In this study, we have examined human theca stem cells [hTSCs] in vitro differentiation capacity into human oocyte like cells [hOLCs]


Materials and Methods: In this interventional experiment study, hTSCs were isolated from the theca layer of small antral follicles [3-5 mm in size]. Isolated hTSCs were expanded and cultured in differentiation medium, containing 5% human follicular fluid, for 50 days. Gene expressions of PRDM1, PRDM14, VASA, DAZL, OCT4, ZP1, 2, 3 GDF9, SCP3 and DMC1 were evaluated by quantitative reverse transcription polymerase chain reaction [qRT-PCR] on days 0, 18, and 25 after monoculture as well as one week after co-culture with human granulosa cells [hGCs]. In addition, GDF9, OCT4, DAZL, VASA, and ZP3 proteins were immune-localized in oocyte-like structures


Results: After 16-18 days, the color of the medium became acidic. After 25 days, the cells started to differentiate into round-shaped cells [20-25 ìm diameter]. One week after co-culturing with hGCs, the size of the round cells increased 60 to70 ìm and convert to hOLCs. However, these growing cells expressed some primordial germ cell [PGC]- and germ cell genes [PRDM1, PRDM14, VASA, DAZL, and OCT4] as well as oocyte specific genes [ZP1, 2, 3 and GDF9], and meiotic-specific markers [SCP3 and DMC1]. In addition, GDF9, OCT4, DAZL, VASA, and ZP3 proteins were present in hOLCs


Conclusion: To sum up, hTSCs have the ability to differentiate into hOLCs. This introduced model paved the way for further in vitro studies of the exact mechanisms behind germ cell formation and differentiation. However, the functionality of hOLCs needs further investigation

12.
Cell Journal [Yakhteh]. 2019; 20 (4): 537-543
in English | IMEMR | ID: emr-199623

ABSTRACT

Objective: A recent innovative approach, based on induction of sublethal oxidative stress to enhance sperm cryosurvival, has been applied before sperm cryopreservation. The purpose of this study was to investigate the effects of different induction times of sublethal oxidative stress before cryopreservation on human post-thawed sperm quality


Materials and Methods: In this experimental study, we selected semen samples [n=20] from normozoospermic men according to 2010 World Health Organization [WHO] guidelines. After processing the samples by the density gradient method, we divided each sample into 5 experimental groups: fresh, control freezing, and 3 groups exposed to 0.01M sodium nitroprusside [SNP] [nitric oxide [NO] donor] for 30 [T30], 60 [T60], or 90 minutes [T90] at 37C and 5% CO2 before cryopreservation. Motion characteristics [computer-assisted sperm analyser], viability, apoptosis [annexin V/propidium iodide [PI] assay], DNA fragmentation [sperm chromatin structure assay [SCSA]], and caspase 3 activity [FLICA Caspase Detection Kit] were assessed after thawing. The results were analysed by using one-way ANOVA and Tukey's test. The means were significantly different at P<0.05


Results: Cryopreservation significantly decreased sperm viability and motility parameters, and increased the percentage of apoptosis, caspase 3 activity, and DNA fragmentation [P<0.01] compared to the fresh group. The T60 group had a higher significant percentage of total motility [TM] and progressive motility compared with other cryopreserved groups [P<0.05]. We observed a significantly lower percentage of apoptotic rate and caspase 3 activity in the T60 group compared to the other cryopreserved groups [P<0.05]. DNA integrity was not significantly affected by this time of sublethal stress induction [P>0.05]


Conclusion: Our results have demonstrated that the application of sublethal oxidative stress by using 0.01 MuM NO for 60 minutes before the freezing process can be a beneficial approach to improve post-thawed human sperm quality

13.
Cell Journal [Yakhteh]. 2019; 20 (4): 544-551
in English | IMEMR | ID: emr-199624

ABSTRACT

Objective: In the present study, we investigated the possible epigenotoxic effect of dimethyl sulfoxide [DMSO] on buffalo fibroblast cells and on reconstructed oocytes during buffalo-bovine interspecies somatic cell nuclear transfer [iSCNT] procedure and its effect on rate and quality of blastocyst which derived from these reconstructed oocytes


Materials and Methods: In this experimental study, cell viability of buffalo fibroblasts was assessed after exposure to various concentration [0.5, 1, 2 and 4%] of DMSO using MTS assay. The epigenetic effect of DMSO was also assessed in terms of DNA methylation in treated cells by flowcytometry. Reconstructed oocytes of buffalo-bovine iSCNT exposed for 16 hours after activation to non-toxic concentration of DMSO [0.5%] to investigate the respective level of 5-methylcytosine, cleavage and blastocyst rates and gene expression [pluripotent genes: OCT4, NANOG, SOX2, and trophectodermal genes: CDX2 and TEAD4] of produced blastocysts


Results: Supplementation of culture medium with 4% DMSO had substantial adverse effect on the cell viability after 24 hours. DMSO, at 2% concentration, affected cell viability after 48 hours and increased DNA methylation and mRNA expression of DNMT3A in fibroblast cells. Exposure of reconstructed oocytes to 0.5% DMSO for 16 hours post activation did not have significant effect on DNA methylation, nor on the developmental competency of reconstructed oocyte, however, it decreased the mRNA expression of NANOG in iSCNT blastocysts


Conclusion: Depending on the dose, DMSO might have epigenotoxic effect on buffalo fibroblast cells and reconstructed oocytes and perturb the mRNA expression of NANOG in iSCNT blastocysts

14.
Cell Journal [Yakhteh]. 2019; 20 (4): 552-558
in English | IMEMR | ID: emr-199625

ABSTRACT

Objective: Over the last years, vitrification has been widely used for oocyte cryopreservation, in animals and humans; however, it frequently causes minor and major epigenetic modifications. The effect of oocyte vitrification on levels of acetylation of histone H4 at lysine 12 [AcH4K12], and histone acetyltransferase [Hat] expression, was previously assessed; however, little is known about the inhibition of Hat expression during oocyte vitrification. This study evaluated the effect of anacardic acid [AA] as a Hat inhibitor on vitrified mouse oocytes


Materials and Methods: In this experimental study, 248 mouse oocytes at metaphase II [MII] stage were divided in three experimental groups namely, fresh control oocytes [which were not affected by vitrification], frozen/thawed oocytes [vitrified] and frozen/thawed oocytes pre-treated with AA [treatment]. Out of 248 oocytes, 173 oocytes were selected and from them, 84 oocytes were vitrified without AA [vitrified group] and 89 oocytes were pretreated with AA, and then vitrified [treatment group]. Fresh MII mouse oocytes were used as control group. Hat expression and AcH4K12 levels were assessed by using real-time quantitative polymerase chain reaction [PCR] and immunofluoresce staining, respectively. In addition, survival rate was determined in vitrified and treatment oocytes


Results: Hat expression and AcH4K12 modification significantly increased [4.17 +/- 1.27 [P.0.001] and 97.57 +/- 6.30 [P<0.001], respectively] in oocytes that were vitrified, compared to the fresh oocytes. After treatment with AA, the Hat mRNA expression and subsequently H4K12 acetylation levels were significantly reduced [0.12 +/- 0.03 [P.0.001] and 89.79 +/- 3.20 [P.0.05], respectively] in comparison to the vitrified group. However, the survival rate was not significantly different between the vitrified [90.47%] and treatment [91.01%] groups [P>0.05]


Conclusion: The present study suggests that AA reduces vitrification risks caused by epigenetic modifications, but does not affect the quality of vitrification. In fact, AA as a Hat inhibitor was effective in reducing the acetylation levels of H4K12

15.
Cell Journal [Yakhteh]. 2019; 20 (4): 559-563
in English | IMEMR | ID: emr-199626

ABSTRACT

Objective: Innate immunity factors are associated with type 2 diabetes [T2DM] and its complications. Therefore, T2DM has been suggested to be an immune-dependent disease. Elevated fasting glucose level and higher concentrations of innate immunity soluble molecules are not only related with insulin resistance, but inflammation is also an important factor in beta cell dysfunction in T2DM. Toll-like receptor 2 [TLR-2], which has an important role in inducing innate immune cells, is thought to have suppressive roles on immune responses in T2DM. We therefore aimed to investigate the possible role of TLR-2 del -196-174 and Arg753Gln variants in T2DM pathogenesis


Materials and Methods: This study was designed as a case-control study. Polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP] technique was used to genotype the two variants in 100 T2DM patients and 98 agematched controls


Results: We found significantly higher frequencies of TLR-2 del -196-174 DD genotype [P=0.003], ID genotype [P=0.009] and D allele [P=0.001] in patients compared with controls. In addition, the II genotype [P=0.001] and the I allele [P=0.003] frequencies were elevated in healthy controls. We did not find any significant differences in frequency distribution for the Arg753Gln variant in study groups


Conclusion: We suggest that carrying the D allele of the TLR-2 del -196-174 variant may be related as a risk factor for T2DM

16.
Cell Journal [Yakhteh]. 2019; 20 (4): 564-568
in English | IMEMR | ID: emr-199627

ABSTRACT

Objective: Considerable research shows that long non-coding RNAs, those longer than 200 nucleotides, are involved in several human diseases such as various cancers and cardiovascular diseases. Their significant role in regulating the function of endothelial cells, smooth muscle cells, macrophages, vascular inflammation, and metabolism indicates the possible effects of lncRNAs on the progression of atherosclerosis which is the most common underlying pathological process responsible for coronary artery disease [CAD]. The aim of present study was to assess whether the expression of the lnc RNA H19 was associated with a susceptibility to CAD by evaluating the expression level of H19 in the peripheral blood


Materials and Methods: A case-control study of 50 CAD patients and 50 age and sex-matched healthy controls was undertaken to investigate whether the H19 lncRNA expression level is associated with a CAD using Taqman Real-Time polymerase chain reaction [PCR]


Results: The subsequent result indicated that the H19 lncRNA was over-expressed in CAD patients in comparison with the controls. However, it was not statistically significant. This overexpression may be involved in coronary artery disease progression


Conclusion: We report here, the up-regulation of H19 lncRNA in the whole blood of CAD patients and suggest a possible role for H19 in the atherosclerosis process and its consideration as novel biomarker for CAD

17.
Cell Journal [Yakhteh]. 2019; 20 (4): 569-575
in English | IMEMR | ID: emr-199628

ABSTRACT

Objective: We sought to apply Shannon's entropy to determine colorectal cancer genes in a microarray dataset


Materials and Methods: In the retrospective study, 36 samples were analysed, 18 colorectal carcinoma and 18 paired normal tissue samples. After identification of the gene fold-changes, we used the entropy theory to identify an effective gene set. These genes were subsequently categorised into homogenous clusters


Results: We assessed 36 tissue samples. The entropy theory was used to select a set of 29 genes from 3128 genes that had fold-changes greater than one, which provided the most information on colorectal cancer. This study shows that all genes fall into a cluster, except for the R08183 gene


Conclusion: This study has identified several genes associated with colon cancer using the entropy method, which were not detected by custom methods. Therefore, we suggest that the entropy theory should be used to identify genes associated with cancers in a microarray dataset

18.
Cell Journal [Yakhteh]. 2019; 20 (4): 576-583
in English | IMEMR | ID: emr-199629

ABSTRACT

Objective: Hemoglobinopathies such as beta-thalassemia and sickle cell disease [SCD] are inherited disorders that are caused by mutations in beta-globin chain. Gamma-globin gene reactivation can ameliorate clinical manifestations of betathalassemia and SCD. Drugs that induce fetal hemoglobin [HbF] can be promising tools for treatment of beta-thalassemia and SCD patients. Recently, it has been shown that Simvastatin [SIM] and Romidepsin [ROM] induce HbF. SIM is a BCL11a inhibitor and ROM is a HDAC inhibitor and both of these drugs are Food and Drug Administration [FDA]-approved for hypercholesterolemia and cutaneous T-cell lymphoma respectively. Our aim was to evaluate the synergistic effects of these drugs in inducing HbF


Materials and Methods: In our experimental study, we isolated CD34+ cells from five cord blood samples that were cultured in erythroid differentiation medium containing ROM and Simvastatin. Then Gamma-globin, BCL11a and HDAC gene expression were evaluated on the 7th and 14th day of erythroid differentiation by real-time polymerase chain reaction [PCR] and immunocytochemistry


Results: Our results showed that combination of SIM and ROM significantly increased Gamma-globin gene expression and inhibit BCL11a and HDAC expression compared to results of using each of them alone. SIM and ROM lead to 3.09- fold increase in HbF production compared to the control group. Also, SIM inhibited BCL11a expression [0.065-fold] and ROM inhibited HDAC1 expression [0.47-fold] as two important inhibitors of HbF production after birth


Conclusion: We propose combination therapy of these drugs may be ameliorate clinical manifestation in beta-thalassemia and SCD with at least side effects and reduce the need for blood transfusion

19.
Cell Journal [Yakhteh]. 2019; 20 (4): 584-591
in English | IMEMR | ID: emr-199630

ABSTRACT

Objective: Substantial effort has been put into designing DNA-based biosensors, which are commonly used to detect presence of known sequences including the quantification of gene expression. Porous silicon [PSi], as a nanostructured base, has been commonly used in the fabrication of optimally transducing biosensors. Given that the function of any PSi-based biosensor is highly dependent on its nanomorphology, we systematically optimized a PSi biosensor based on reflectometric interference spectroscopy [RIS] detecting the high penetrance breast cancer susceptibility gene, BRCA1


Materials and Methods: In this experimental study, PSi pore sizes on the PSi surface were controlled for optimum filling with DNA oligonucleotides and surface roughness was optimized for obtaining higher resolution RIS patterns. In addition, the influence of two different organic electrolyte mixtures on the formation and morphology of the pores, based on various current densities and etching times on doped p-type silicon, were examined. Moreover, we introduce two cleaning processes which can efficiently remove the undesirable outer parasitic layer created during PSi formation. Results of all the optimization steps were observed by field emission scanning electron microscopy [FE-SEM]


Results: DNA sensing reached its optimum when PSi was formed in a two-step process in the ethanol electrolyte accompanied by removal of the parasitic layer in NaOH solution. These optimal conditions, which result in pore sizes of approximately 20 nm as well as a low surface roughness, provide a considerable RIS shift upon complementary sequence hybridization, suggesting efficient detectability


Conclusion: We demonstrate that the optimal conditions identified here makes PSi an attractive solid-phase DNA-based biosensing method and may be used to not only detect full complementary DNA sequences, but it may also be used for detecting point mutations such as single nucleotide substitutions and indels

20.
Cell Journal [Yakhteh]. 2019; 20 (4): 592-598
in English | IMEMR | ID: emr-199631

ABSTRACT

Objective: Amyotrophic lateral sclerosis [ALS] is the most severe disorder within the spectrum of motor neuron diseases [MND] that has no effective treatment and a progressively fatal outcome. We have conducted two clinical trials to assess the safety and feasibility of intravenous [IV] and intrathecal [IT] injections of bone marrow derived mesenchymal stromal cells [BM-MSCs] in patients with ALS


Materials and Methods: This is an interventional/experimental study. We enrolled 14 patients that met the following inclusion criteria: definitive diagnosis of sporadic ALS, ALS Functional Rating Scale [ALS-FRS] .24, and .40% predicted forced vital capacity [FVC]. All patients underwent bone marrow [BM] aspiration to obtain an adequate sample for cell isolation and culture. Patients in group 1 [n=6] received an IV and patients in group 2 [n=8] received an IT injection of the cell suspension. All patients in both groups were followed at 24 hours and 2, 4, 6, and 12 months after the injection with ALS-FRS, FVC, laboratory tests, check list of side effects and brain/spinal cord magnetic resonance imaging [MRI]. In each group, one patient was lost to follow up one month after cell injection and one patient from IV group died due to severe respiratory insufficiency and infection


Results: During the follow up there were no reports of adverse events in terms of clinical and laboratory assessments. In MRI, there was not any new abnormal finding. The ALS-FRS score and FVC percentage significantly reduced in all patients from both groups


Conclusion: This study has shown that IV and IT transplantation of BM-derived stromal cells is safe and feasible [Registration numbers: NCT01759797 and NCT01771640]

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