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Nanomedicine has emerged as a promising therapeutic approach, but its translation to the clinic has been hindered by the lack of cellular models to anticipate how tumor cells will respond to therapy. Three-dimensional (3D) cell culture models are thought to more accurately recapitulate key features of primary tumors than two-dimensional (2D) cultures. Heterotypic 3D tumor spheroids, composed of multiple cell types, have become more popular than homotypic spheroids, which consist of a single cell type, as a superior model for mimicking in vivo tumor heterogeneity and physiology. The stromal interactions demonstrated in heterotypic 3D tumor spheroids can affect various aspects, including response to therapy, cancer progression, nanomedicine penetration, and drug resistance. Accordingly, to design more effective anticancer nanomedicinal therapeutics, not only tumor cells but also stromal cells (e.g., fibroblasts and immune cells) should be considered to create a more physiologically relevant in vivo microenvironment. This review aims to demonstrate current knowledge of heterotypic 3D tumor spheroids in cancer research, to illustrate current advances in utilizing these tumor models as a novel and versatile platform for in vitro evaluation of nanomedicine-based therapeutics in cancer research, and to discuss challenges, guidelines, and future directions in this field.
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Nanomedicina , Esferoides Celulares , Linhagem Celular Tumoral , FibroblastosRESUMO
Objectives: Autism is a complicated neurodevelopmental disorder characterized by social interaction deficiencies, hyperactivity, anxiety, communication disorders, and a limited range of interests. The zebrafish (Danio rerio) is a social vertebrate used as a biomedical research model to understand social behavior mechanisms. Materials and Methods: After spawning, the eggs were exposed to sodium valproate for 48 hr, after which the eggs were divided into eight groups. Except for the positive and control groups, there were six treatment groups based on oxytocin concentration (25, 50, and 100 µM) and time point (24 and 48 hr). Treatment was performed on days 6 and 7, examined by labeling oxytocin with fluorescein-5-isothiocyanate (FITC) and imaging with confocal microscopy and the expression levels of potential genes associated with the qPCR technique. Behavioral studies, including light-dark background preference test, shoaling behavior, mirror test, and social preference, were performed on 10, 11, 12, and 13 days post fertilization (dpf), respectively. Results: The results showed that the most significant effect of oxytocin was at the concentration of 50 µM and the time point of 48 hr. Increased expression of shank3a, shank3b, and oxytocin receptor genes was also significant at this oxytocin concentration. Light-dark background preference results showed that oxytocin in the concentration of 50 µM significantly increased the number of crosses between dark and light areas compared with valproic acid (positive group). Also, oxytocin showed an increase in the frequency and time of contact between the two larvae. We showed a decrease in the distance in the larval group and an increase in time spent at a distance of one centimeter from the mirror. Conclusion: Our findings showed that the increased gene expression of shank3a, shank3b, and oxytocin receptors improved autistic behavior. Based on this study some indications showed that oxytocin administration in the larval stage could significantly improve the autism-like spectrum.
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BACKGROUND: Oral targeted small molecules, including sphingosine 1 phosphate receptor (S1PR) modulators and tyrosine kinase inhibitors (TKIs), seem to revolutionize the management of inflammatory bowel disease (IBD). To select the most effective treatment, there is an unmet need to comparatively study their mechanism of action, efficacy, and toxicity in the preclinical stage and further translate it into clinical practice. METHODS: Using 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced adult zebrafish colitis model, LC50 of fingolimod and tofacitinib were determined based on the acute toxicity test to compare aquatic toxicity potential. Subsequently, the efficacy of different concentrations of tofacitinib and fingolimod was compared using flow cytometry, qPCR, and histopathology analyses. RESULTS: TNBS significantly reduced the length of villi, and the number of goblet cells increased the level of TNF-α, MyD88, and NF-κB2, the thickness of villi and necrosis, and induced histopathological changes. All of these parameters were reversed almost dose-dependently with both medications, with the highest concentration of fingolimod being superior to other groups. Additionally, results from qPCR analysis suggested that these medications might suppress canonical and non-canonical NF-κB pathways by targeting toll-like receptors and MyD88. LC50 of tofacitinib and fingolimod was 0.9014 and 0.36 mg/L, respectively. Hence, both are in the cory 1 of the Global Harmonization System (GHS) aquatic toxicity and are toxic to adult zebrafish life. CONCLUSION: Given the better efficacy of fingolimod, it is worth translating the effectiveness and safety of S1PR modulators into IBD patients and comparing them with TKIs in head-to-head studies; albeit, their toxicity should not be overlooked.
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Colite , Doenças Inflamatórias Intestinais , Animais , Cloridrato de Fingolimode/uso terapêutico , Cloridrato de Fingolimode/efeitos adversos , Peixe-Zebra/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , Colite/induzido quimicamente , Ácido Trinitrobenzenossulfônico , Doenças Inflamatórias Intestinais/tratamento farmacológicoRESUMO
Enhancers are distal cis-acting elements that are commonly recognized to regulate gene expression via cooperation with promoters. Along with regulating gene expression, enhancers can be transcribed and generate a class of non-coding RNAs called enhancer RNAs (eRNAs). The current discovery of abundant tissue-specific transcription of enhancers in various diseases such as cancers raises questions about the potential role of eRNAs in disease diagnosis and therapy. This review aimed to demonstrate the current understanding of eRNAs in cancer research with a focus on the potential roles of eRNAs as prognostic and diagnostic biomarkers in cancers.
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Gene therapy is regarded as a valuable strategy for efficient cancer treatment. However, the design of effective delivery systems that can deliver gene materials such as siRNA specifically to the tumour tissues plays a pivotal role in cancer therapy. For this reason, a targeted cationic liposome for melanoma treatment was developed. This system consists of cyclic RGD peptide conjugated to DSPE-PEG2000, cholesterol, DOTAP and DSPC as cationic and neutral lipids, respectively. Cyclic RGD was selected based on speculation that cyclic RGD would effectively transport anti-signal transducer and activator of transcription 3 (STAT3) siRNA into melanoma cell via integrin receptors. The prepared liposomes provided excellent stability against electrolyte and serum nucleases. Targeted liposomes remarkably exhibited higher cellular internalisation in comparison with the non-targeted system in flow cytometry and confocal microscopy. Furthermore, incorporating peptide on the surface of liposomes resulted in considerably high cytotoxicity, a 2.1-times raise in apoptosis induction, and a significantly enhanced STAT3 gene suppression as compared with the corresponding non-targeted formulation on B16F10 murine melanoma cells. Whole-body imaging confirmed the more significant tumour accumulation of targeted liposomes in B16F10 melanoma xenograft tumour-bearing mice. Consequently, c-RGD peptide modified liposome suggests a promising option for specific siRNA delivery into melanoma cells.
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Lipossomos , Melanoma , Animais , Cátions , Linhagem Celular Tumoral , Humanos , Melanoma/tratamento farmacológico , Camundongos , Oligopeptídeos , Peptídeos , Peptídeos Cíclicos , RNA Interferente PequenoRESUMO
Colorectal cancer (CRC) is one of the most malignant cancer types, characterized by elevated mortality rate and treatment resistance. Despite the progress achieved in the explanation of the molecular basis of the disease as well as introducing potential biomarkers in the clinical practice, further investigation is essential to identify innovative molecules that contribute to colorectal carcinogenesis. Circular RNAs (circRNAs) are a novel and unexplored RNA type, associated with various human pathological conditions. Recently, circRNAs have been identified to be enriched and stable in exosomes and can exert their functions when exosomes reach neighboring or distant cells. Increasing evidence indicates that these so called exosomal circRNAs (exo-circRNAs) act as signaling molecules to regulate cancer proliferation, metastasis, and sensitivity to radio- and chemotherapy. This review aims to discuss the latest progress in exo-circRNAs studies in CRC with an emphasis on their potential as promising diagnostic molecular markers and therapeutic targets.
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BACKGROUND: The continuous accessibility of local animals for sustainable use is being eroded annually. Thus, a strategic vision for the conservation of biodiversity is of far-reaching emphasis to deal with unprecedented challenges in the local population extension facing in the future. This study aimed to establish and cryopreserve endangered Markhoz goat (Capra hircus) fibroblast cell lines in vitro. METHODS AND RESULTS: These primary fibroblast cells were isolated from 58 Iranian Markhoz goats and individually cultured by explant technique in DMEM medium supplemented with 10% FBS and 2 mM L-Glutamine, in the presence of Penicillin (200 U/ml)-Streptomycin (200 mg/ml) during the first passage number. The extracted cell lines were confirmed morphologically as fibroblast cells. The population doubling time for DMEM-cultured cells was 23 ± 0.5 h. Chromosomal analysis indicated a total chromosome number of 2n = 60 with > 95% frequency. The cultured cells were checked for bacteria, fungi, yeast, and mycoplasma contaminations and the results were reported negative. The efficiencies of the fluorescent protein encoded by VSV-G (pMDG) and lentiviral pCSGW vectors reported in a range of 65% value. According to the species identification analysis, the goat cell lines were banked and confirmed without any miss- and cross-contamination. CONCLUSIONS: The significant issue in this paper can be concluded about the first report of the establishment of endangered Markhoz goat cell banking inside the country. This study demonstrated the successful establishment of a genetically stable fibroblast bank as a valuable genetic resource for the endangered Iranian Markhoz goat breed.
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Conservação dos Recursos Naturais/métodos , Criopreservação/métodos , Espécies em Perigo de Extinção , Fibroblastos , Cabras/genética , Animais , Cruzamento/métodos , Técnicas de Cultura de Células , Linhagem Celular , Cromossomos/genética , Irã (Geográfico) , Cariótipo , Cariotipagem/métodos , Mycoplasma/genética , Reação em Cadeia da Polimerase/métodosRESUMO
Accurate relative gene expression analysis by reverse transcription-quantitative polymerase chain reaction relies on the usage of suitable reference genes for data normalization. The RNA content of small extracellular vesicles including exosomes is growingly considered as cancer biomarkers. So, reliable relative quantification of exosomal messenger RNA (mRNA) is essential for cancer diagnosis and prognosis applications. However, suitable reference genes for accurate normalization of a target gene in exosomes derived from cancer cells are not depicted yet. Here, we analyzed the expression and stability of eight well-known reference genes namely GAPDH, B2M, HPRT1, ACTB, YWHAZ, UBC, RNA18S, and TBP in exosomes-isolated from the liver (Huh7, HepG2, PLC/PRF/5) and breast (SK-BR-3) cancer cell lines using five different algorithms including geNorm, BestKeeper, Delta Ct, NormFinder, and RefFinder. Our results showed that ACTB, TBP, and HPRT1 were not expressed in exosomes-isolated from studied liver and breast cancer cell lines. The geNorm and BestKeeper algorithms indicated GAPDH and UBC as the most stable candidates. Moreover, Delta Ct and NormFinder algorithms showed YWHAZ as the most stable reference genes. Comprehensive ranking calculated by the RefFinder algorithm also pointed out GAPDH, YWHAZ, and UBC as the first three stable reference genes. Taken together, this study validated the common reference genes stability in exosomal mRNA derived from liver and breast cancer cell lines for the first time. We believe that this study would be the first step in finding more stable reference genes in exosomes that triggers more accurate detection of exosomal biomarkers.
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Perfilação da Expressão Gênica/métodos , Genes Essenciais/genética , Estabilidade de RNA/genética , Proteínas 14-3-3/genética , Algoritmos , Mama/patologia , Neoplasias da Mama/genética , Linhagem Celular , Exossomos/genética , Feminino , Expressão Gênica/genética , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/genética , Humanos , Fígado/patologia , Neoplasias Hepáticas/genética , Estabilidade de RNA/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Padrões de Referência , Ubiquitina C/genéticaRESUMO
AIMS: Exosomes hold great promise as bio-inspired delivery vehicles. Mesenchymal stem cells (MSCs) are recognized for their potential to yield huge quantities of exosomes. We aimed to investigate the potential use of modified exosomes derived from genetically modified dental pulp MSCs (DPSCs) as a carrier to deliver tumor suppressor miR-34a to repress proliferation of breast carcinoma cells. MATERIALS AND METHODS: miR-34a-overexpressing DPSCs were prepared using XMIRXpress-34a lentivectors. The anticancer effects of the miR-34a-loaded exosomes were evaluated on breast carcinoma cells through apoptosis, migration, and invasion assays. Given the structural similarity between exosomes and liposomes, we compared the exosome-mediated miRNA delivery efficiency with that of liposomes. KEY FINDINGS: Our data demonstrated that genetically modified DPSCs were capable of secretion of exosomes enriched with therapeutic miRNAs and presented the feasibility of application of exosome-based vehicle for gene delivery. SIGNIFICANCE: We showed the potential of MSC-derived exosomes as a tool for delivery of miRNAs in vitro. Nevertheless, optimizing gene-loading approaches is required before exosomes can be intended as a miRNA carrier for therapeutic applications.
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Neoplasias da Mama/terapia , Polpa Dentária/citologia , Sistemas de Liberação de Medicamentos , Exossomos/genética , Células-Tronco Mesenquimais/citologia , MicroRNAs/administração & dosagem , Transplante de Células-Tronco , Apoptose , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Movimento Celular , Proliferação de Células , Polpa Dentária/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , Células Tumorais CultivadasRESUMO
Bisphenol A (BPA) as a pervasive endocrine-disrupting compound (EDC) has been shown to cause multiple detrimental effects including cardiovascular disorders, pregnancy complications, obesity, glucose metabolism disorders, and reproductive toxicity even at a concentration as low as tolerable daily intake (TDI) (4 µg/kg/day). In the present study, a novel ultra-sensitive, electrochemical aptasensor was designed using a screen-printed carbon electrode (SPCE) modified by gold nanoparticles (Au NPs) conjugated to thiolated aptamers for accurate determination of BPA in biological, industrial and environmental samples. To characterize the electrochemical properties of the aptasensor, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were implemented. Detection of BPA was also performed through differential pulse voltammetry (DPV) in [Fe(CN)6]3-/4- electrolyte solution. Under optimum condition, the present electrochemical aptasensor demonstrated an outstanding linear response in the concentration range of 1 pM to 10 nM with a remarkably low limit of detection of 0.113 pM. Due to the superb affinity between anti-BPA aptamers and BPA molecules, the designed aptasensor did not show any significant interaction with other analytes in real samples. Also, fabricated biosensor remained perfectly stable in long-term storage. The analytical results of the fabricated aptasensor are well compatible with those obtained by the ELISA method, indicating the trustworthiness and reasonable accuracy of the application of aptasensor in real samples. Overall, the proposed aptasensor would be a credible and economical method of precise, reproducible, and highly selective detection of minimum levels of BPA in food containers and clinical samples. This would be a promising strategy to enhance the safety of food products and reduce the risk of BPA daily exposure.
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Oocyte maturation is an important phase in fertility and any disorder in this process could lead to infertility. The most common disorder during folliculogenesis is polycystic ovary syndrome (PCOS). Due to the secretive activity of granulosa cells (GCs), they play a vital role in folliculogenesis. Although scientists use various cellular and molecular methods to have a better understanding of the mechanism of these cells, some limitations still exist in GC culture such as low primary cell yield and proliferation capability. Therefore, immortalization of primary cells is an approach to overcome these limitations. In the current study, GCs were obtained from two females, one with PCOS and one with normal folliculogenesis. In the first stage, we established two human GC (hGC) lines by immortalizing them through retrovirus-mediated transfer of the human telomerase reverse transcriptase (hTERT) and c-Myc genes. Subsequently, the normal and PCOS cell lines were characterized and were investigated for their growth features. The cell lines were also examined in terms of immortal markers of hTERT, follicle stimulating hormone receptor (FSHR), aromatase, anti-Müllerian hormone (AMH), growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), estrogen, and progesterone. Our results indicated that the normal and PCOS cell lines both showed similar characteristics to GCs during the follicular stage in normal and PCOS women. The normal and PCOS cell lines demonstrate molecular mechanisms similar to that of GCs such as folliculogenesis, oogenesis, and steroidogenesis, which enable researchers to perform further investigations in future.
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Endocrine-disrupting chemicals (EDCs) such as bisphenol A (BPA), which is widely used in the plastic industry, have recently been considered to be involved in the pathogenesis of metabolic disorders, including obesity and diabetes. The present study aimed to examine the potentially detrimental effects of BPA on glucose and energy metabolism at the epigenetic level. The blood glucose profile of Wistar rats receiving different oral doses of BPA over 28 days was assessed. At the end of the treatment, the islets of Langerhans were isolated and purified, and their RNA content was extracted. MicroRNA (miRNA) profiling was evaluated using the next generation sequencing (NGS) method. After performing bioinformatic analysis of the NGS data, the gene ontology and data enrichment in terms of significantly disturbed miRNAs were evaluated through different databases, including Enrichr and DIANA tools. Additionally, the DNA methylation and the level of expression of two critical genes in glucose metabolism (PPARγ, Pdx1) were assessed. Subchronic BPA exposure (406 mg/kg/day) disturbed the blood glucose profile (fasting blood glucose and oral glucose tolerance) of Wistar rats and resulted in considerable cytotoxicity. NGS data analyses revealed that the expression of some crucial miRNAs involved in ß-cell metabolism and diabetes occurrence and development, including miR-375, miR-676, miR-126-a, and miR-340-5p, was significantly disrupted. According to the DNA methylation evaluation, both PPARγ and Pdx1 genes underwent changes in the methylation level at particular loci on the gene's promoter. The expression levels of these genes were upregulated and downregulated, respectively. Overall, subchronic BPA exposure could cause epigenetic dysregulation at the gene level and interfere with the expression of key miRNAs and the methylation process of genes involved in glucose homeostasis. Understanding the exact underlying mechanisms by which BPA and other EDCs induce endocrine disturbance could be of great importance in the way of finding new preventive and therapeutic approaches.
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Compostos Benzidrílicos/farmacologia , Epigênese Genética/efeitos dos fármacos , Ilhotas Pancreáticas/efeitos dos fármacos , Fenóis/farmacologia , Administração Oral , Animais , Compostos Benzidrílicos/administração & dosagem , Biologia Computacional , Metilação de DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Epigênese Genética/genética , Ilhotas Pancreáticas/metabolismo , Masculino , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Fenóis/administração & dosagem , Ratos , Ratos WistarRESUMO
Erythropoietin (EPO), a hematopoietic factor, is one of the promising neuroprotective candidates in neurodegenerative disorders such as Alzheimer's disease (AD). Due to the high molecular weight, hydrophilicity and rapid clearance from circulation, EPO could not completely pass the blood-brain barrier in the case of systemic administration. To overcome this limitation, EPO-loaded Solid Lipid Nanoparticle (EPO-SLN) was developed in this study using a double emulsion solvent evaporation method (W1/O/W2). Glycerin monostearate (GMS), span®80/span®60, Dichloromethane (DCM) and tween®80 were chosen as lipid, internal phase surfactants, solvent, and external aqueous phase surfactant, respectively. After physicochemical evaluations, the effect of EPO-SLN on the beta-amyloid-induced AD-like animal model was investigated. In vivo evaluations, it was demonstrated that the memory was significantly restored in cognitive deficit rats treated with EPO-SLN compared to the rats treated with native drug using the Morris water maze test. In addition, EPO-SLN reduced the oxidative stress, ADP/ATP ratio, and beta-amyloid plaque deposition in the hippocampus more effectively than the free EPO. Hence, the designed SLN can be regarded as a promising system for safe and effective delivery of EPO in the AD.
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Doença de Alzheimer/tratamento farmacológico , Eritropoetina/uso terapêutico , Aprendizagem em Labirinto/efeitos dos fármacos , Transtornos da Memória/tratamento farmacológico , Nanopartículas/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Doença de Alzheimer/induzido quimicamente , Peptídeos beta-Amiloides , Animais , Modelos Animais de Doenças , Eritropoetina/administração & dosagem , Hipocampo/efeitos dos fármacos , Masculino , Nanopartículas/administração & dosagem , Fármacos Neuroprotetores/farmacologia , Ratos , Ratos Wistar , Resultado do TratamentoRESUMO
Mesenchymal stem cells (MSCs) are multipotent stromal cells present in various adult tissues. Several studies suggest that MSCs secrete exosomes that perform as mediators in the tumor niche and play several roles in tumorigenesis, angiogenesis, and metastasis. In contrast, there are other studies supporting the tumor-suppressing effects of MSC-derived exosomes. Therefore, the exact association of MSC exosomes and tumor cells remains open to debate. This review aimed to demonstrate the present knowledge of MSC-derived exosomes in cancer research and to illustrate current approaches to make use of modified exosomes as a platform in therapeutic strategies in cancer.
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Exossomos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Neoplasias/terapia , Animais , Ensaios Clínicos como Assunto , Sistemas de Liberação de Medicamentos , Humanos , Neoplasias/irrigação sanguínea , Neovascularização Patológica/terapiaRESUMO
Solid lipid nanoparticle (SLN) is a promising approach for delivery of various drugs including proteins and peptides. However, the loading of hydrophilic drugs into the lipoid matrix of SLNs is challenging. The statistical design is a potential method facilitating the optimization of nanoparticles characteristics. In this study, the Box-Behnken design was conducted to optimize the preparation of Erythropoietin (EPO) loaded SLNs. Circular dichroism, size exclusion chromatography, SDS-PAGE, and ELISA tests were used to prove the compatibility of the process with the stability of EPO. In the controlled situation, EPO preserved its conformation and activity during the SLN preparation. Regarding the particle size, entrapment efficiency, and polydispersity index, an optimum formulation was obtained with 130 mg Span®80, 152.5 µl EPO, and 1.9 min high-shear homogenization. Using the optimum condition, 280 nm sized SLNs with the narrow size distribution of 0.282 and entrapment efficiency of 43.4% were acquired. The in vitro cytotoxicity of optimum SLN formulation was conducted using MTT assay to show its safety on the evaluated cell line. The in vivo studies demonstrated that 2500 U EPO loaded SLN has similar or even better effects on elevating the RBC, hemoglobin, and hematocrit level compared to the 5000 U EPO solution. Generally, this study proposed a suitable EPO-loaded SLN preparation method as a potential drug delivery system for proteins.
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Sistemas de Liberação de Medicamentos/métodos , Eritropoetina/química , Lipídeos/química , Nanopartículas/química , Cromatografia em Gel , Dicroísmo Circular , Portadores de Fármacos/química , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção EnzimáticaRESUMO
Some of lizard species have the ability to lose their tail in order to defend against predators and regenerate the new tail. Lizard's regenerated tail has attracted scientists' attention for unraveling the regeneration process, but less information is known about the cellular characterization and cell growth properties of original tail. This research aimed to report cell culture and banking process of rough-tailed gecko or Cyrtopodion scabrum's original tail cell sample from inner tissue without skin using tissue explant technique. For banking reports, it is essential to analyze this cells' potential to proliferate, to investigate biological aspects such as cell culture features, differentiation and chromosome number and to report its species identification and quality control. To achieve optimal growth conditions, three different temperatures for incubation including 18, 23 and 37 °C and two different media including DMEM and L-15 were applied. The expanded cells were studied for their potential to adipose and osteoblast differentiation. Results indicated that lizard's original tail cells could be successfully obtained by explant technique. The cells demonstrated fibroblast like morphology with population doubling times of approximately 24 ± 0.5 h. Karyotyping analysis showed a distribution of 2n = 40 chromosome number for this cell line. The comparison of different incubation media and temperatures showed that cell growth is equally optimal in all mentioned conditions according to growth curves. Adipose and osteoblast differentiation was obviously observed in these cells which confirms the hint of stem-ness in the produced mixed cells. According to cell banking policies, produced cells were also checked for bacterial, fungal, yeast and mycoplasma contaminations and no contamination was observed. Multiplex PCR for identification of species confirmed the species of lizard with no cross-contamination with other cells in the cell bank. Establishment of authenticated and well-characterized lizard's original tail cell line will provide a valuable source for subsequent in vitro regenerative research and molecular studies which are not feasible in in vivo methods. This finding will allow us to get an opportunity to create and preserve a new collection of lizard cell lines in the future.
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Identification of animal species is one of the major concerns in food regulatory control and quality assurance system. Different approaches have been used for species identification in animal origin of feedstuff. This study aimed to develop a multiplex PCR approach to detect the origin of meat and meat products. Specific primers were designed based on the conserved region of mitochondrial Cytochrome C Oxidase subunit I (COX1) gene. This method could successfully distinguish the origin of the pig, camel, sheep, donkey, goat, cow, and chicken in one single reaction. Since PCR products derived from each species represent unique molecular weight, the amplified products could be identified by electrophoresis and analyzed based on their size. Due to the synchronized amplification of segments within a single PCR reaction, multiplex PCR is considered to be a simple, fast, and inexpensive technique that can be applied for identification of meat products in food industries. Nowadays, this technique has been considered as a practical method to identify the species origin, which could further applied for animal feedstuffs identification.
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OBJECTIVES: Alzheimer's disease (AD) is one of the most common forms of neurodegenerative diseases. Despite vast ongoing researches focusing on the area, little is known about novel treatments. In this study, we aimed to survey the effects of Capparis spinosa (C. spinosa) extract on amyloid-beta peptide (Aß)-injected rat. METHODS: For this purpose, hydroalcoholic extracts of caper leaf and fruit were prepared. Total phenolic content, DPPH, and FRAP assay were accomplished to determine antioxidant activity of C. spinosa. HPLC analysis was conducted to measure rutin and quercetin content of selected parts of the plant. Higher levels of flavonoids were observed in leaves of the plant. Twelve male Wistar Aß-induced rats were randomly divided in four groups of (1) Aß-/DW+: Sham-operated group (2) Aß+/DW+: Aß-injected group (3) Aß+/RU+: Standard rutin treatment (4) Aß+/CS+: C. spinosa extract treatment. After 6 weeks of oral administration, real-time qPCR were conducted to determine APP, BACE-1, PSEN-1, and PSEN-2 genes expression in the hippocampus of rats. RESULTS: HPLC analysis showed high levels of rutin and quercetin in leaves of Capparis. Rutin was 16939.2 ± 0.01 and quercetin was 908.93 ± 0.01â µg/g fresh weight. In fruit, 1019.52 ± 0.01 rutin and 97.86 ± 0.01â µg/g FW quercetin were measured. Expression of BACE-1, APP, PSEN-1, and PSEN-2 genes in comparison with the control group showed significant down regulation. DISCUSSION: Results of the study demonstrated that C. spinosa has the potential to down regulate inflammation-involved genes in AD, due to its high levels of flavonoids and could be beneficial as a dietary complement in AD patients.
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Doença de Alzheimer/genética , Capparis/química , Flavonoides/farmacologia , Extratos Vegetais/farmacologia , Doença de Alzheimer/tratamento farmacológico , Peptídeos beta-Amiloides , Animais , Frutas/química , Masculino , Folhas de Planta/química , Quercetina/farmacologia , Ratos , Ratos Wistar , Rutina/farmacologiaRESUMO
Discovery of biosensors has acquired utmost importance in the field of healthcare. Recent advances in biological techniques and instrumentation involving nanomaterials, surface plasmon resonance, and aptasensors have developed innovative biosensors over classical methods. Integrated approaches provided a better perspective for developing specific and sensitive devices with wide potential applications. Type 2 diabetes mellitus is a complex disease affecting almost every tissue and organ system, with metabolic complications extending far beyond impaired glucose metabolism. Although there is no known cure for Type 2 diabetes, early diagnosis and interventions are critical to prevent this disease and can postpone or even prevent the serious complications that are associated with diabetes. Biomarkers for type 2 diabetes are useful for prediction and intervention of the disease at earlier stages. Proper selection of biomarkers that represent health and disease states is vital for disease diagnosis and treatment by detecting it before it manifests. In this respect, we provide an overview of different types of biosensors being used, ranging from electrochemical, fluorescence-based, nanomonitors, SPR-based, and field-effect transistor biosensors for early detection and management of diabetes with focus on prediabetes. In the future, novel non-invasive technologies combined with blood and tissue-based biomarkers will enable the detection, prevention, and treatment of diabetes and its complications long before overt disease develops.
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Biomarcadores/metabolismo , Técnicas Biossensoriais , Diabetes Mellitus Tipo 2/diagnóstico , Diagnóstico Precoce , Diabetes Mellitus Tipo 2/metabolismo , Humanos , Resistência à Insulina/genética , Ressonância de Plasmônio de SuperfícieRESUMO
Corneal haze, commonly caused by deep physical and chemical injuries, can greatly impair vision. Growth factors facilitate fibroblast proliferation and differentiation, which leads to haze intensity. In this study, the potential effect of chitosan (CS) and thiolated-chitosan (TCS) nanoparticles and solutions on inhibition of fibroblast proliferation, fibroblast to myofibroblast differentiation, neovascularization, extracellular matrix (ECM) deposition, and pro-fibrotic cytokine expression was examined. Transforming growth factor beta-1 (TGFß1) was induced by interleukin-6 (IL6) in human corneal fibroblasts and expression levels of TGFß1, Platelet-derived growth factor (PDGF), α-smooth muscle actins (α-SMA), collagen type I (Col I), fibronectin (Fn) and vascular endothelial growth factor (VEGF) were quantified using qRT-PCR. To assess wound-healing capacity, TCS-treated mice were examined for α-SMA positive cells, collagen deposition, inflammatory cells and neovascularization through pathological immunohistochemistry. The results revealed that CS and TCS could down-regulate the expression levels of TGFß1 and PDGF comparable to that of TGFß1 knockdown experiment. However, down-regulation of TGFß1 was not regulated through miR29b induction. Neovascularization along with α-SMA and ECM deposition were significantly diminished. According to these findings, CS and TCS can be considered as potential anti-fibrotic and anti-angiogenic therapeutics. Furthermore, TCS, thiolated derivative of CS, will increase mucoadhesion of the polymer at the corneal surface which makes the polymer efficient and non-toxic therapeutic approach for corneal injuries.