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RNA Expression Analysis of Mycobacterial Methyltransferases Genes in Different Resistant Strains of Mycobacterium tuberculosis. 结核分枝杆菌甲基转移酶基因在不同耐药菌株中的RNA表达分析
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-05-01 DOI: 10.52547/ibj.26.3.240
Samira Tarashi, Mohammad Saber Zamani, Golnaz Bahramali, Andrea Fuso, Farzam Vaziri, Morteza Karimipoor, Abolfazl Fateh, Seyed Davar Siadat

Background: Tuberculosis infection still represents a global health issue affecting patients worldwide. Strategies for its control may be not as effective as it should be, specifically in case of resistant strains of Mycobacterium tuberculosis (M.tb.) In this regard, the role of mycobacterial methyltransferases (MTases) in TB infection can be fundamental, though it has not been broadly deciphered.

Methods: Five resistant isolates of M.tb were obtained. M.tb H37Rv (ATCC 27249) was used as a reference strain. Seven putative mycobacterial MTase genes (Rv0645c, Rv2966c, Rv1988, Rv1694, Rv3919c, Rv2756c, and Rv3263) and Rv1392 as SAM synthase were selected for analysis. PCR-sequencing and qRT-PCR were performed to compare mutations and expression levels of MTases in different strains. The 2-ΔΔCt method was employed to calculate the relative expression levels of these genes.

Results: Only two mutations were found in isoniazid resistance (INHR) strain for Rv3919c (T to G in codon 341) and Rv1392 (G to A in codon 97) genes. Overexpression of Rv0645c, Rv2756c, Rv3263, and Rv2966c was detected in all sensitive and resistant isolates. However, Rv1988 and Rv3919c decreased and Rv1694 increased in the sensitive strains. The Rv1392 expression level also decreased in INHR isolate.

Conclusion: We found a correlation between mycobacterial MTases expression and resistance to antibiotics in M.tb strains. Some MTases undeniably are virulence factors that specifically hijack the host defense mechanism. Further evaluations are needed to explore the complete impact of mycobacterial MTases within specific strains of M.tb to introduce novel diagnosis and treatment strategies.

背景:结核病感染仍然是一个影响全世界患者的全球性健康问题。控制结核分枝杆菌的策略可能不像应有的那样有效,特别是在耐药结核分枝杆菌菌株的情况下。在这方面,分枝杆菌mtase在结核感染中的作用可能是根本的,尽管它还没有被广泛破译。方法:获得5株耐药结核分枝杆菌。以结核分枝杆菌H37Rv (ATCC 27249)作为参考菌株。选取7个分枝杆菌MTase基因(Rv0645c、Rv2966c、Rv1988、Rv1694、Rv3919c、Rv2756c、Rv3263)和Rv1392作为SAM合成酶进行分析。采用pcr测序和qRT-PCR比较不同菌株mtase的突变和表达水平。2-ΔΔCt法计算这些基因的相对表达量。结果:在INHR株中仅发现Rv3919c(密码子341从T到G)和Rv1392(密码子97从G到A)两个基因突变。在所有敏感和耐药菌株中均检测到Rv0645c、Rv2756c、Rv3263和Rv2966c过表达。而Rv1988和Rv3919c在敏感株中呈下降趋势,Rv1694呈上升趋势。Rv1392在INHR分离株中的表达水平也有所下降。结论:结核分枝杆菌mtase表达与耐药之间存在相关性。不可否认,一些mtase是特异性劫持宿主防御机制的毒力因子。需要进一步的评估来探索分枝杆菌mtase在特定结核分枝杆菌菌株中的全面影响,以引入新的诊断和治疗策略。
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引用次数: 0
Effects of Nanofiber Scaffolds Coated with Nanoparticulate and Microparticulate Freeze Dried Bone Allograft on the Morphology, Adhesion, and Proliferation of Human Mesenchymal Stem Cells 纳米颗粒和微颗粒冻干同种异体骨包被纳米纤维支架对人间充质干细胞形态、粘附和增殖的影响
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-04-27 DOI: 10.52547/ibj.26.3.193
Shabnam Aghayan, E. Seyedjafari, Shadi Hamidi
Background: Freeze dried bone allograft nanoparticles on a nanofiber membrane may serve as an ideal scaffold for bone regeneration. This study aimed to assess the biological behavior of human MSCs in terms of proliferation and adhesion to nanoparticulate and microparticulate FDBA scaffolds on PLLA nanofiber membrane. Methods: In this experimental study, PLLA nanofiber scaffolds were synthesized by the electrospinning method. The FDBA nanoparticles were synthesized mechanically. The FDBA nanoparticles and microparticles were loaded on the surface of PLLA nanofiber membrane. A total of 64 scaffold samples in four groups of n-FDBA/PLLA, FDBA/PLLA, PLLA and control were placed in 24-well polystyrene tissue culture plates; 16 wells were allocated to each group. Data were analyzed using one-way ANOVA and Bonferroni test. Results: The proliferation rate of MSCs was significantly higher in the nanoparticulate group compared to the microparticulate group at five days (p = 0.034). Assessment of cell morphology at 24 hours revealed spindle-shaped cells with a higher number of appendages in the nanoparticulate group compared to other groups. Conclusion: MSCs on n-FDBA/PLLA scaffold were morphologically more active and flatter with a higher number of cellular appendages, as compared to FDBA/PLLA. It seems that the nanoparticulate scaffold is superior to the microparticulate scaffold in terms of proliferation, attachment, and morphology of MSCs in vitro.
背景:纳米纤维膜上冷冻干燥的同种异体骨纳米颗粒可能是骨再生的理想支架。本研究旨在评估人骨髓间充质干细胞在增殖和粘附于PLLA纳米纤维膜上的纳米和微粒FDBA支架方面的生物学行为。方法:采用静电纺丝法合成PLLA纳米纤维支架。采用机械法制备了FDBA纳米粒子。将FDBA纳米粒子和微粒负载在PLLA纳米纤维膜表面。将n-FDBA/PLLA、FDBA/PLA、PLLA和对照四组共64个支架样品置于24孔聚苯乙烯组织培养板中;每组分配16口井。数据采用单因素方差分析和Bonferroni检验进行分析。结果:与微粒组相比,纳米颗粒组的MSCs在第五天的增殖率显著更高(p=0.034)。24小时的细胞形态评估显示,与其他组相比,纳米粒子组的纺锤形细胞具有更多的附属物。结论:与FDBA/PLLA相比,n-FDBA/PLA支架上的MSCs在形态上更活跃、更平坦,具有更多的细胞附属物。在体外培养的MSCs的增殖、附着和形态方面,纳米颗粒支架似乎优于微粒支架。
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引用次数: 0
Regulatory Effect of let-7f Transfection in Non-Small Cell Lung Cancer on its Candidate Target Genes let-7f转染非小细胞肺癌癌症对候选靶基因的调控作用
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-04-27 DOI: 10.52547/ibj.26.3.209
V. Zafari, M. Asadi, Nasim Bakhtiyari, Mahsa Sadeghzadeh, M. Khalili, H. Zarredar, Soghra Bornehdeli, E. Seyedrezazadeh
Background: Let-7f has essential impacts on biological processes; however, its biological and molecular functions in lung cancer pathogenesis have yet been remained unclear. We aimed to investigate the expression level of let-7f and its candidate target genes both in lung cancer tissues and A549 cell line. Methods: Bioinformatics databases were first used to select candidate target genes of let-7f. Then the relative gene and protein expressions of let-7f and its target genes, including HMGA2, ARID3B, SMARCAD1, and FZD3, were measured in lung tissues of Non-Small Cell Lung Cancer (NSCLC) patients and A549 cell line using quantitative real-time PCR and Western blotting. The electroporation method was used to transfect A549 cells with let-7f mimic and microRNA inhibitor. The impact of let-7f transfection on the viability of A549 cells was assessed using MTT assay. The expression data of studied genes were analyzed statistically Results: Results indicated significant downregulated expression level of let-7f-5p (p = 0.0013) and upregulated level of the HMGA2 and FZD3 in NSCLC cases (p < 0.05). In A549 cells, after transfection with let-7f mimic, the expression of both mRNA and protein levels of HMGA2, ARID3B, SMARCAD1, and FZD3 decreased. Also, the overexpression of let-7f significantly inhibited the A549 cell proliferation and viability (p = 0.017). Conclusion: Our findings exhibited the high value of let-7f and HMGA2 as biomarkers for NSCLC. The let-7f, as a major tumor suppressor regulatory factor via direct targeting genes (e.g. HMGA2), inhibits lung cancer cell viability and proliferation and could serve as a marker for the early diagnostic of NSCLC.
背景:Let-7f对生物过程具有重要影响;然而,其在癌症发病机制中的生物学和分子功能尚不清楚。我们旨在研究let-7f及其候选靶基因在癌症组织和A549细胞系中的表达水平。方法:首先利用生物信息学数据库筛选let-7f的候选靶基因。然后,使用定量实时PCR和Western印迹法测量let-7f及其靶基因(包括HMGA2、ARID3B、SMARCAD1和FZD3)在非小细胞肺癌癌症(NSCLC)患者的肺组织和A549细胞系中的相对基因和蛋白质表达。采用电穿孔法用let-7f模拟物和microRNA抑制剂转染A549细胞。用MTT法评估let-7f转染对A549细胞活力的影响。结果:let-7f-5p的表达水平在NSCLC患者中显著下调(p=0.0013),HMGA2和FZD3的表达水平显著上调(p<0.05)。在A549细胞中,用let-7f模拟物转染后,HMGA1、ARID3B、SMARCAD1和FZD3 mRNA和蛋白水平均下降。此外,let-7f的过表达显著抑制了A549细胞的增殖和活力(p=0.017)。结论:let-7f和HMGA2作为NSCLC的生物标志物具有很高的价值。let-7f作为一种主要的肿瘤抑制调节因子,通过直接靶向基因(如HMGA2)抑制癌症细胞的生存能力和增殖,并可作为NSCLC早期诊断的标志物。
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引用次数: 1
Reporting Two Novel Mutations in Two Iranian Families with Cystic Fibrosis, Molecular and Bioinformatic Analysis 报道两个伊朗囊性纤维化家族中的两个新突变,分子和生物信息学分析
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-04-26 DOI: 10.52547/ibj.3713
Amin Hosseini Nami, M. Kabiri, S. Zeinali
Background: Cystic fibrosis is the most common heredity disease among the Caucasian population. More than 350 known pathogenic variations in the CFTR gene (NM_000492.4) cause CF. Herein, we report the outcome of our investigation in two unrelated Iranian families with CF patients. Methods: We conducted phenotypic examination, segregation, linkage analysis, and CFTR gene sequencing to define causative mutations. Results: We found two novel mutations in the present study. The first one was a deletion causing frameshift, c.299delT p.(Leu100Profs*7), and the second one was a missense mutation, c.1857G>T, at nucleotide binding domain 1 of the CFTR protein. Haplotype segregation data supported our new mutation findings. Conclusion: Findings of this study expand the spectrum of CFTR pathogenic variations and can improve prenatal diagnosis and genetic counseling for CF.
背景:囊性纤维化是高加索人群中最常见的遗传性疾病。超过350种已知的CFTR基因(NM_000492.4)致病性变异导致CF。在此,我们报告了我们对两个无血缘关系的伊朗CF患者家庭的调查结果。方法:通过表型检查、分离、连锁分析和CFTR基因测序来确定致病突变。结果:我们在本研究中发现了两个新的突变。第一个是导致移码的缺失,c.299delT p.(Leu100Profs*7),第二个是CFTR蛋白核苷酸结合域1的错义突变,c.1857G>T。单倍型分离数据支持我们新的突变发现。结论:本研究结果扩大了CFTR致病变异谱,可改善CF的产前诊断和遗传咨询。
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引用次数: 1
Electrophysiological Recordings from Embryonic Mouse Motoneurons Cultured on Electrospun Poly-Lactic Acid (PLA) and Polypyrrole-Coated PLA Scaffolds 聚乳酸(PLA)和聚吡咯包被PLA支架培养小鼠胚胎运动神经元的电生理记录
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-04-03 DOI: 10.52547/ibj.26.3.183
Esmeralda Zuñiga, Odin Ramírez, Carlos Martínez
Background: Biomaterials used as cell growth stimulants should be able to provide adequate cell adhesion with no alteration in cell function. In this work, we developed a 3D model of mouse spinal cord motoneurons on scaffolds composed of electrospun PLA fibers and plasma-polymerized PPy-coated PLA fibers. Methods: The functionality of the cultured motoneurons was assessed by evaluating both the electrophysiological response (i.e., the whole-cell Na+ and K+ currents and the firing of action potentials) and also the expression of the VAChaT by immunostaining techniques. While the expression of the VAChaT was confirmed on motoneurons cultured on the fibrous scaffolds, the electrophysiological responses indicated Na+ and K+ currents with lower amplitude and slower action potentials when compared to the response recorded from spinal cord motoneurons cultured on Poly-DL-Ornithine/Laminin- and plasma-polymerized PPy-coated coverslips. Results: From a morphological viewpoint, motoneurons cultured on PLA and PPy-coated PLA scaffolds did not show the development of dendritic and/or axonal processes, which were satisfactorily observed in the bidimensional cultures. Conclusion: We hypothesize that the apparently limited development of dendritic and/or axonal processes could produce a deleterious effect on the electrophysiological response of the cells, which might be due to the limited growth surface available in the fibrous scaffolds and/or to an undesired effect of the purification process.
背景:用作细胞生长刺激剂的生物材料应该能够在不改变细胞功能的情况下提供足够的细胞粘附。在这项工作中,我们在由电纺PLA纤维和等离子体聚合PPy涂层PLA纤维组成的支架上开发了小鼠脊髓运动神经元的3D模型。方法:通过免疫染色技术评估电生理反应(即全细胞Na+和K+电流以及动作电位的激发)和VAChaT的表达来评估培养的运动神经元的功能。虽然VAChaT在纤维支架上培养的运动神经元上的表达得到了证实,但与在聚DL-鸟氨酸/层粘连蛋白和血浆聚合PPy涂层盖玻片上培养的脊髓运动神经元记录的反应相比,电生理反应显示Na+和K+电流具有较低的振幅和较慢的动作电位。结果:从形态学角度来看,在PLA和PPy涂层的PLA支架上培养的运动神经元没有显示出树突和/或轴突突起的发育,这在二维培养中得到了令人满意的观察。结论:我们假设树突和/或轴突突起明显有限的发育可能会对细胞的电生理反应产生有害影响,这可能是由于纤维支架中可用的生长表面有限和/或纯化过程的不良影响。
{"title":"Electrophysiological Recordings from Embryonic Mouse Motoneurons Cultured on Electrospun Poly-Lactic Acid (PLA) and Polypyrrole-Coated PLA Scaffolds","authors":"Esmeralda Zuñiga, Odin Ramírez, Carlos Martínez","doi":"10.52547/ibj.26.3.183","DOIUrl":"https://doi.org/10.52547/ibj.26.3.183","url":null,"abstract":"Background: Biomaterials used as cell growth stimulants should be able to provide adequate cell adhesion with no alteration in cell function. In this work, we developed a 3D model of mouse spinal cord motoneurons on scaffolds composed of electrospun PLA fibers and plasma-polymerized PPy-coated PLA fibers. Methods: The functionality of the cultured motoneurons was assessed by evaluating both the electrophysiological response (i.e., the whole-cell Na+ and K+ currents and the firing of action potentials) and also the expression of the VAChaT by immunostaining techniques. While the expression of the VAChaT was confirmed on motoneurons cultured on the fibrous scaffolds, the electrophysiological responses indicated Na+ and K+ currents with lower amplitude and slower action potentials when compared to the response recorded from spinal cord motoneurons cultured on Poly-DL-Ornithine/Laminin- and plasma-polymerized PPy-coated coverslips. Results: From a morphological viewpoint, motoneurons cultured on PLA and PPy-coated PLA scaffolds did not show the development of dendritic and/or axonal processes, which were satisfactorily observed in the bidimensional cultures. Conclusion: We hypothesize that the apparently limited development of dendritic and/or axonal processes could produce a deleterious effect on the electrophysiological response of the cells, which might be due to the limited growth surface available in the fibrous scaffolds and/or to an undesired effect of the purification process.","PeriodicalId":14500,"journal":{"name":"Iranian Biomedical Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41876100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Evaluation of the Prognostic Value of CD56 (140 kDa Isoform) Expression in Breast Cancer Tissues: an Eight-Year Retrospective Study 乳腺癌组织中CD56 (140 kDa亚型)表达的预后价值评估:一项8年回顾性研究
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-04-03 DOI: 10.52547/ibj.26.3.175
Kianoush Niknam, A. Safaei, A. Ghaderi
Background: Identification of specific antigens is highly beneficial for early detection, diagnosis, staging, and outcome prediction of cancer. This study aimed to evaluate the expression and prognostic value of CD56 (140 kDa isoform) in IDC. Methods: Sixty-five patients with IDC who underwent radical surgery or mastectomy as the primary treatment were included. Proper formalin-fixed and paraffin embedded tissue blocks of the patients were prepared and stained by IHC for CD56 (140 kDa isoform) molecule. Chi-square and fisher exact tests were used to compare the results against the clinicopathologic data of patients. Kaplan-Meier and log-rank test were employed to study the prognostic value of the target antigen. Results: The expression pattern of CD56 was granular and cytoplasmic. There were significant associations between the intensity of CD56 expression in invasive cells and carcinoma in situ (p = 0.005) and normal ducts (p = 0.010). Among all clinicipathologic parameters, there was only a significant association between the expression of ER and CD56 (p = 0.023). Neither OS (p = 0.356) nor DFS (p = 0.976) had significant correlation with CD56 expression. Conclusion: Our data indicated that the CD56 marker offers no prognostic value in terms of predicting the OS or DFS for up to eight years after primary surgery. Furthermore, the intensity of its expression is similar between normal, non-invasive, and invasive cells. Considering the generally better outcome of ER+ BC patients than their ER-counterparts, the CD56 marker may be indirectly associated with a more favorable prognosis among IDC patients.
背景:特异性抗原的鉴定对癌症的早期发现、诊断、分期和预后预测非常有益。本研究旨在评估CD56 (140 kDa亚型)在IDC中的表达及其预后价值。方法:65例以根治性手术或乳房切除术为主要治疗方法的IDC患者。制备合适的福尔马林固定组织块和石蜡包埋组织块,免疫组化染色CD56 (140 kDa亚型)分子。采用卡方检验和fisher精确检验将结果与患者的临床病理资料进行比较。采用Kaplan-Meier检验和log-rank检验研究靶抗原的预后价值。结果:CD56的表达呈颗粒状和细胞质状。浸润细胞中CD56表达强度与原位癌(p = 0.005)和正常导管(p = 0.010)有显著相关性。在所有临床病理参数中,只有ER与CD56表达有显著相关性(p = 0.023)。OS (p = 0.356)和DFS (p = 0.976)与CD56表达均无显著相关性。结论:我们的数据表明,CD56标志物在预测原发性手术后长达8年的OS或DFS方面没有预后价值。此外,其表达强度在正常、非侵袭性和侵袭性细胞之间相似。考虑到ER+ BC患者的预后通常比ER+ BC患者好,CD56标记物可能与IDC患者更有利的预后间接相关。
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引用次数: 0
A Green Approach for the Biosynthesis of Gold Nanoparticles Using Cuminum cyminum L. Seed and Its Application for Pain Management in Rats 利用菊芋种子生物合成金纳米粒子的绿色方法及其在大鼠疼痛管理中的应用
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-14 DOI: 10.52547/ibj.26.3.219
S. Golabi, M. Adelipour, A. Mohammadi, Kosar Omidian, A. Rastqar, M. Naghashpour
Background: This study investigated the antinociceptive effect of cumin and its biosynthesized AuNPs. Methods: Cumin extract (E) and cumin-AuNPs (GN) were prepared and administered intraperitoneally at the concentrations of 200, 500, and 1000 mg/ml to 27 male rats. UV–Vis spectroscopy and AFM were applied for AuNPs synthesis confirmation. The nociceptive behavior was assessed, and IL-6 serum levels were measured. Results: Cumin-AuNPs showed a peak absorption of 515 nm, and a size of about 40 nm. Three different concentrations of extract had no significant effect on acute and chronic nociceptive behavior. GN + E200 (46.00 ± 10.59) showed a significant acute anti-nociceptive effect compared to the control (98.66 ± 4.91; p = 0.029) and SS300 (98.33 ± 20.30; p = 0.029) groups. Also, GN + E500 (42.00 ± 11.84) significantly reduced acute nociceptive behavior compared to the control (98.66 ± 4.91; p = 0.019), SS300 (98.33 ± 20.30; p = 0.020), and GN + E1000 (91.00 ± 26.00; p = 0.040) groups. IL-6 serum levels reduced significantly in GN + E500 (24.65 ± 10.38; p = 0.002) and SS300 (33.08 ± 1.68; p = 0.039) compared to the controls (46.24 ± 3.02). Chronic nociceptive behavior was significantly lower in the SS300 (255.33 ± 26.30) compared to E200 (477.00 ± 47.29; p = 0.021), E500 (496.25 ± 46.29; p = 0.013), and GN + E500 (437.00 ± 118.03; p = 0.032) groups. Conclusion: Our findings suggest the potential effects of cumin-AuNPs on formalin-induced nociceptive behavior, which is independent of IL-6serum levels.
背景:本研究探讨了孜然及其生物合成AuNPs的抗伤性作用。方法:制备孜然提取物(E)和孜然- aunps (GN),分别以200、500、1000 mg/ml的浓度腹腔注射27只雄性大鼠。利用紫外可见光谱和原子力显微镜对AuNPs的合成进行了验证。评估伤害性行为,测定血清IL-6水平。结果:孜然- aunps的吸收峰为515 nm,尺寸约为40 nm。三种不同浓度的提取物对急性和慢性伤害性行为均无显著影响。GN + E200(46.00±10.59)较对照组(98.66±4.91)有显著的急性抗伤害性作用;p = 0.029)和SS300(98.33±20.30;P = 0.029)组。GN + E500(42.00±11.84)显著降低了急性伤害性行为,而对照组(98.66±4.91;p = 0.019), SS300(98.33±20.30;p = 0.020), GN + E1000(91.00±26.00;P = 0.040)组。GN + E500组血清IL-6水平显著降低(24.65±10.38;p = 0.002)和SS300(33.08±1.68;P = 0.039),高于对照组(46.24±3.02)。SS300组的慢性伤害性行为(255.33±26.30)显著低于E200组(477.00±47.29);p = 0.021), E500(496.25±46.29;p = 0.013), GN + E500(437.00±118.03;P = 0.032)组。结论:我们的研究结果表明孜然- aunps对福尔马林诱导的伤害性行为的潜在影响与il -6血清水平无关。
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引用次数: 0
Bacterial DNA Detection in the Blood of Healthy Subjects 健康人血液细菌DNA检测
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-13 DOI: 10.52547/ibj.26.3.230
Javad Raeisi, M. Oloomi, M. Zolfaghari, S. Siadat, Mohsen Zargar, Z. Pourramezan
Background: The presence of microbiome in the blood samples of healthy individuals has been addressed. However, no information can be found on the healthy human blood microbiome of Iranian subjects. The current study is thus aimed to investigate the existence of bacteria or bacterial DNA in healthy individuals. Methods: Blood samples of healthy subjects were incubated in BHI broth at 37 °C for 72 h. The 16S rRNA PCR and sequencing were performed to analyze bacterial isolates. The 16S rRNA PCR was directly carried out on DNA samples extracted from the blood of healthy individuals. NGS was conducted on blood samples with culture-positive results. Results: Fifty blood samples were tested, and six samples were positive by culture as confirmed by Gram staining and microscopy. The obtained 16S rRNA sequences of cultured bacterial isolates revealed the presence of Bacilli and Staphylococcus species by clustering in the GeneBank database (≥97% identity). The 16S rRNA gene sequencing results of one non-cultured blood specimen showed the presence of Burkholderia. NGS results illustrated the presence of Romboutsia, Lactobacillus, Streptococcus, Bacteroides, and Staphylococcus in the blood samples of positive cultures. Conclusion: The dormant blood microbiome of healthy individuals may give the idea that the steady transfer of bacteria into the blood does not necessarily lead to sepsis. However, the origins and identities of blood-associated bacterial rDNA sequences need more evaluation in the healthy population.
背景:健康个体血液样本中微生物组的存在已经得到了解决。然而,没有找到关于伊朗受试者健康人体血液微生物组的信息。因此,目前的研究旨在调查健康个体中细菌或细菌DNA的存在。方法:健康人血液标本在BHI肉汤中37℃孵育72 h,采用16S rRNA PCR和测序对分离细菌进行分析。直接对健康人血液中提取的DNA样本进行16S rRNA PCR。对培养阳性的血样进行NGS检测。结果:共检测50份血样,革兰氏染色及镜检证实培养阳性6份。培养菌株的16S rRNA序列在GeneBank数据库中聚类,发现Bacilli和Staphylococcus两种细菌存在(一致性≥97%)。一份非培养血液标本的16S rRNA基因测序结果显示存在伯克氏菌。NGS结果显示阳性培养的血液样本中存在Romboutsia, Lactobacillus, Streptococcus, Bacteroides和葡萄球菌。结论:健康个体的休眠血液微生物组可能提示细菌稳定转移到血液中并不一定导致败血症。然而,血液相关细菌rDNA序列的起源和特性需要在健康人群中进行更多的评估。
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引用次数: 3
Fabrication of Silk Scaffold Containing Simvastatin-Loaded Silk Fibroin Nanoparticles for Regenerating Bone Defects 含辛伐他汀负载丝素纳米颗粒的骨缺损再生丝支架的制备
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-01 DOI: 10.52547/ibj.26.2.116
Fatemeh Mottaghitalab, Hamidreza Motasadizadeh, Mohammad Ali Shokrgozar, Shahrokh Shojaei, Mehdi Farokhi

Background: In the present study, a tissue engineered silk fibroin (SF) scaffold containing simvastatin-loaded silk fibroin nanoparticles (SFNPs) were used to stimulate the regeneration of the defected bone.

Methods: At first, the porous SF scaffold was prepared using freeze-drying. Then simvastatin-loaded SFNPs were made by dissolvation method and embedded in the SF scaffold. Afterwards, the scaffold and the NPs were characterized in terms of physicochemical properties and the ability to release the simvastatin small molecule.

Results: The results exhibited that the SF scaffold had a porous structure suitable for releasing the small molecule and inducing the proliferation and attachment of osteoblast cells. SFNPs containing simvastatin had spherical morphology and were 174 ± 4 nm in size with -24.5 zeta potential. Simvastatin was also successfully encapsulated within the SFNPs with 68% encapsulation efficiency. Moreover, the small molecule revealed a sustained release profile from the NPs during 35 days. The results obtained from the in vitro cell-based studies indicated that simvastatin-loaded SFNPs embedded in the scaffold had acceptable capacity to promote the proliferation and alkaline phosphatase production of osteoblast cells while inducing osteogenic matrix precipitation.

Conclusion: The SF scaffold containing simvastatin-loaded SFNPs could have a good potential to be used as a bone tissue-engineered construct.

背景:在本研究中,组织工程丝素(SF)支架含有辛伐他汀负载的丝素纳米颗粒(SFNPs)来刺激缺损骨的再生。方法:首先采用冷冻干燥法制备多孔SF支架。然后采用溶解法制备辛伐他汀负载的SFNPs,并包埋于SF支架中。然后,对支架和NPs的理化性质和释放辛伐他汀小分子的能力进行表征。结果:SF支架具有适合小分子释放和诱导成骨细胞增殖附着的多孔结构。含辛伐他汀的SFNPs呈球形,尺寸为174±4 nm, zeta电位为-24.5。辛伐他汀也被成功地包封在SFNPs中,包封率为68%。此外,小分子在35天内显示出NPs的持续释放特征。体外细胞实验结果表明,负载辛伐他汀的SFNPs包埋在支架中,具有促进成骨细胞增殖和碱性磷酸酶产生的能力,同时诱导成骨基质沉淀。结论:含有辛伐他汀负载的SFNPs的SF支架具有良好的骨组织工程构建潜力。
{"title":"Fabrication of Silk Scaffold Containing Simvastatin-Loaded Silk Fibroin Nanoparticles for Regenerating Bone Defects","authors":"Fatemeh Mottaghitalab,&nbsp;Hamidreza Motasadizadeh,&nbsp;Mohammad Ali Shokrgozar,&nbsp;Shahrokh Shojaei,&nbsp;Mehdi Farokhi","doi":"10.52547/ibj.26.2.116","DOIUrl":"https://doi.org/10.52547/ibj.26.2.116","url":null,"abstract":"<p><strong>Background: </strong>In the present study, a tissue engineered silk fibroin (SF) scaffold containing simvastatin-loaded silk fibroin nanoparticles (SFNPs) were used to stimulate the regeneration of the defected bone.</p><p><strong>Methods: </strong>At first, the porous SF scaffold was prepared using freeze-drying. Then simvastatin-loaded SFNPs were made by dissolvation method and embedded in the SF scaffold. Afterwards, the scaffold and the NPs were characterized in terms of physicochemical properties and the ability to release the simvastatin small molecule.</p><p><strong>Results: </strong>The results exhibited that the SF scaffold had a porous structure suitable for releasing the small molecule and inducing the proliferation and attachment of osteoblast cells. SFNPs containing simvastatin had spherical morphology and were 174 ± 4 nm in size with -24.5 zeta potential. Simvastatin was also successfully encapsulated within the SFNPs with 68% encapsulation efficiency. Moreover, the small molecule revealed a sustained release profile from the NPs during 35 days. The results obtained from the in vitro cell-based studies indicated that simvastatin-loaded SFNPs embedded in the scaffold had acceptable capacity to promote the proliferation and alkaline phosphatase production of osteoblast cells while inducing osteogenic matrix precipitation.</p><p><strong>Conclusion: </strong>The SF scaffold containing simvastatin-loaded SFNPs could have a good potential to be used as a bone tissue-engineered construct.</p>","PeriodicalId":14500,"journal":{"name":"Iranian Biomedical Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8987414/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39953985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
A Historic Review of the Role of CD4+ T-Cell Subsets in Development of the Immune Responses against Cutaneous and Visceral Leishmaniases CD4+ t细胞亚群在皮肤和内脏利什曼病免疫反应中作用的历史回顾
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-01 DOI: 10.52547/ibj.26.2.99
Mohammad Hossein Alimohmmadian, Soheila Ajdary, Fariborz Bahrami

The heterogeneity of CD4+ T cells has been investigated since the late 1970s, when their Th1 and Th2 subsets were coined. Later studies on the cutaneous form of the Leishmaniasis were focused on the experimental models of Leishmania major infection using the susceptible BALB/c and the resistant C57BL/6 mice. At the early 21st century, the regulatory T-cells subpopulation was introduced and its role in concomitant immunity, responsible for lifelong resistance of the host to the reinfection was proposed. Subsequent studies, mainly focused on the visceral form of the infection pointed to the role of IL-17, produced by Th17 subset of CD4+ T cells that along the neutrophils were shown to have important yet equivocal functions in protection against or exacerbation of the infection. Altogether, the current knowledge indicates that the above four subsets could orchestrate the immune, the regulatory and the inflammatory responses of the host against different forms of leishmaniases.

自20世纪70年代末CD4+ T细胞的Th1和Th2亚群被创造出来以来,人们就开始研究它们的异质性。后来对利什曼病皮肤形式的研究主要集中在利什曼病大感染的实验模型上,使用易感的BALB/c和耐药的C57BL/6小鼠。在21世纪初,调节性t细胞亚群被引入,其在伴随免疫中的作用被提出,负责宿主对再感染的终身抵抗。随后的研究,主要集中在感染的内脏形式,指出IL-17的作用,由CD4+ T细胞的Th17亚群产生,沿着中性粒细胞被证明在保护或加剧感染方面具有重要但模糊的功能。总的来说,目前的知识表明,上述四个亚群可以协调宿主对不同形式的利什曼病的免疫、调节和炎症反应。
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引用次数: 1
期刊
Iranian Biomedical Journal
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