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1.
BMC Complement Med Ther ; 24(1): 234, 2024 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-38879474

RESUMEN

BACKGROUND: This study explored the impact of predicted miRNAs on DNA methyltransferases (DNMTs) and the PODXL gene in Nalm6 cells, revealing the significance of these miRNAs in acute lymphocytic leukemia (ALL). METHODS: A comprehensive approach was adopted, integrating bioinformatic analyses encompassing protein structure prediction, molecular docking, dynamics, and ADMET profiling, in conjunction with evaluations of gene and miRNA expression patterns. This methodology was employed to elucidate the therapeutic potential of catechin compounds in modulating the activity of DNA methyltransferases (DNMTs) and the PODXL gene. RESULTS: The findings from our investigation indicate that catechins possess the capability to inhibit DNMT enzymes. This inhibitory effect is associated with the upregulation of microRNAs miR-200c and miR-548 and a concurrent downregulation of PODXL gene expression. These molecular interactions culminate in an augmented apoptotic response within ALL (Nalm6) cells. CONCLUSION: The study posits that catechins may represent a viable therapeutic avenue for inducing apoptosis in ALL cells. This is achieved through the modulation of epigenetic mechanisms and alterations in gene expression profiles, highlighting the potential of catechins as agents for cancer therapy.


Asunto(s)
Catequina , MicroARNs , Catequina/farmacología , Catequina/análogos & derivados , MicroARNs/genética , Humanos , Línea Celular Tumoral , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Simulación del Acoplamiento Molecular , Metilasas de Modificación del ADN/metabolismo , Simulación por Computador , Apoptosis/efectos de los fármacos
2.
Iran J Basic Med Sci ; 26(6): 708-716, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37275753

RESUMEN

Objectives: The main objective of the current assay was to evaluate the antibacterial and regenerative effects of hydrogel nanocomposite containing pure natural zeolite (clinoptilolite) integrated with alginate (Alg) as wound healing/dressing biomaterials. Materials and Methods: The zeolites were size excluded, characterized by SEM, DLS, XRD, FTIR, and XRF, and then integrated into Alg hydrogel followed by calcium chloride crosslinking. The Alg and alginate zeolite (Alg/Zeo) hydrogel was characterized by swelling and weight loss tests, also the antibacterial, hemocompatibility, and cell viability tests were performed. In animal studies, the burn wound was induced on the back of rats and treated with the following groups: control, Alg hydrogel, and Alg/Zeo hydrogel. Results: The results showed that the hydrodynamic diameter of zeolites was 367 ± 0.2 nm. Zeolites did not show any significant antibacterial effect, however, the hydrogel nanocomposite containing zeolite had proper swelling as well as hemocompatibility and no cytotoxicity was observed. Following the creation of a third-degree burn wound on the back of rats, the results indicated that the Alg hydrogel and Alg/Zeo nanocomposite accelerated the wound healing process compared with the control group. Re-epithelialization, granulation tissue thickness, collagenization, inflammatory cell recruitment, and angiogenesis level were not significantly different between Alg and Alg/Zeo nanocomposite. Conclusion: These findings revealed that although the incorporation of zeolites did not induce a significant beneficial effect in comparison with Alg hydrogel, using zeolite capacity in hydrogel for loading the antibiotics or other effective compounds can be considered a promising wound dressing.

3.
J Cancer Res Ther ; 19(2): 327-334, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37313910

RESUMEN

Objective: Propolis is a viscous resinous honeybee-produced substance with numerous medicinal functions; its composition and texture varies according to the geographic location. It is considered to be a promising natural source for the management and prevention of various pathological conditions. Although several studies have exhibited the anti-cancer activity of different types of propolis, the tumor-suppressing potential of Kermanian propolis against leukemia cell lines has remained poorly understood. Therefore, the current experiment was aimed to reveal the anti-tumor activity of this bioactive compound both as monotherapy and combined therapy with cytarabine against an acute myeloid leukemia (AML) cell line, NB4. Materials and Methods: Following the treatment of NB4 cells with either Kermanian propolis (5, 10, 20, 40, 80, 160, and 320 µg/mL), cytarabine (0.1, 0.25, 0.5, 0.75, 1, and 2 mM), or their combination (40 and 80 µg/mL of Kermanian propolis along with 0.1, 0.25, and 0.5 mM of cytarabine), colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was employed to measure the viability (%) of the cells. Next, to examine the apoptotic rate and the pattern of corresponding gene expression (Bcl-2, Bax, p53, and p21), Annexin-V/PI staining by flow cytometry and quantitative Real-Time polymerase chain reaction assays were performed, respectively. Results: We perceived significant apoptosis induction in a dose-dependent manner following the treatment with Kermanian propolis, cytarabine, and also their combination in the NB4 cell line. In addition, the combined treatment was associated with lower expression of the anti-apoptotic gene (Bcl-2) and higher expression of the pro-apoptotic genes (p53, Bax, and p21) in comparison to mono treatments. Conclusion: The synergistic anti-tumor activity induced by the combination of Kermanian propolis and cytarabine presents a novel and encouraging option for AML treatment.


Asunto(s)
Leucemia Mieloide Aguda , Própolis , Abejas , Humanos , Animales , Regulación hacia Arriba , Própolis/farmacología , Proteína X Asociada a bcl-2/genética , Proteína p53 Supresora de Tumor/genética , Leucemia Mieloide Aguda/tratamiento farmacológico , Apoptosis , Línea Celular , Citarabina
4.
Indian J Hematol Blood Transfus ; 38(2): 373-380, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-35496977

RESUMEN

The present study aims to investigate the changes in different parameters related to the storage time of red blood cell (RBC) units. Microscopic, flow cytometric, and electrophoretic assessments were employed every few days for 60 days to investigate the alterations in morphology, size, phosphatidylserine (PS) externalization, and membrane proteins over time. Morphological transformation from discocytes to spherocytes progressed as the storage time increased, which was accompanied by an increment of cellular size. However, this storage period did not result in the externalization of significant amounts of PS (p > 0.05). Mean Fluorescence Intensity (MFI) values increased by 11% to 23% between days 21 and 35 compared to the day 1 sample (p < 0.001). By day 60, the MFI decreased to about 70% of the day 1 sample. The analysis of membrane proteins' distribution showed a significant drop in band 3 expression after 35 days (p < 0.05 and 0.001 on days 42 and 60, respectively); however, no significant change was observed up to five weeks (p > 0.05). The inconsistency observed between Eosin-5-Maleimide (5-EMA) binding and the relative band 3 content could be due to additional accessibility of 5-EMA to hidden domains of other membrane proteins on RBCs as a result of increased mean corpuscular volume (MCV) and changes in morphology. Overall, our present study represents a step-wise and time-dependent series of events that progressively affects stored RBCs.

5.
Vet Res Forum ; 12(2): 197-202, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34345386

RESUMEN

There has been a prevailing trend in the application of herbal medicine as cancer therapeutics. Calotropis procera is an ayurvedic plant applied to ameliorate various illnesses. There is no report on the anti-tumor effects of the root of the plant on canine tumors, although it has been used for the treatment of various diseases in human medicine. The objective of the present study was to investigate the antitumor potential of ethanolic root extract of C. procera against canine mammary tumor cell line (CF41-Mg). MTT, western blot, and flow cytometry assays were carried out to evaluate the possible cytotoxicity and apoptosis induction of the extract. MTT results showed that the extract had a potent cytotoxic activity in a dose-dependent manner with an IC50 of 9.00 µg mL-1. Based on the results of flow cytometry and western blotting, IC50 concentration of the extract induced significant apoptosis in the studied cell line, possibly through down-regulation of Bcl-2 expression. The results of the present study clearly indicated that the root extract of C. procera had promising anti-cancer activity and could be considered as a candidate for the treatment of mammary tumors.

6.
Genes Environ ; 43(1): 36, 2021 Aug 11.
Artículo en Inglés | MEDLINE | ID: mdl-34380574

RESUMEN

BACKGROUND: Pentavalent antimonial compounds are currently used to treat leishmaniasis and resistance to these drugs is a serious problem. Multidrug resistance protein is an efflux pump of the cell membrane that expels foreign compounds. This study designed to evaluate the mutations in the multi-drug resistance 1 (MDR1) gene, in biopsy specimens of Leishmania tropica, with high resolution melting (HRM) method. In this experimental study, genomic DNA was extracted from 130 patients with skin leishmaniasis. Then, nucleotide changes were investigated throughout the gene using HRM and sequencing methods. The samples categorized in 5 groups by differences in the melting temperature (Tm). RESULT: The nucleotide changes analysis showed that 61% of the samples of different groups that were unresponsive to drug had mutations in the MDR1 gene, which were also confirmed by the sequencing method. These mutations can be one of the factors responsible for non-responsiveness to the treatment. CONCLUSION: According to the findings, it seems that mutation in MDR1 gene could be responsible for drug resistance to pentavalent antimonial compounds. Furthermore, HRM method can be used to diagnose drug resistance in leishmaniasis. It is also recommended that further studies be done regarding the importance of drug resistance in the leishmania affected patients.

7.
Transfus Apher Sci ; 60(5): 103188, 2021 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-34144875

RESUMEN

OBJECTIVES AND BACKGROUND: In December 2019, the first case of COVID-19 was reported in Wuhan, China. Its causative virus, is a novel strain of RNA viruses with high mortality rate. There is no definitive treatment, but among available approaches the use of recovered patients' plasma containing specific antibodies can enhance the immune response against coronavirus. However, the dearth of eligible donors and also ABO incompatibility in plasma transfusion, have limited this therapeutic method. Therefore, it is highly desirable to introduce a simple procedure that allows efficient reduction or even removal of natural ABO antibodies. Accordingly, we aimed to evaluate a RBC-mediated adsorption technique that reduces the titer of the mentioned antibodies in plasma. METHODS/MATERIALS: This experimental study was conducted in Kerman University of Medical Sciences, Kerman, Iran. The pre- and post-incubation antibody titers of 168 plasma samples were determined. For incubation, each plasma sample was exposed (60 min) to different percentages of RBCs at room temperature or 4 °C. RESULTS: The results evidenced that both the concentration of RBCs and temperature had significant decreasing effects on antibody titer (P < 0.001) and all concentrations significantly reduced titer. Compared to RT, 4 °C further reduced the antibody titer. Overall, the best incubation condition for reducing antibody titer in all blood groups was 4 °C and 2% RBCs concentration. CONCLUSION: The presented adsorption procedure is able to produce universal plasma (we call it Ubiquitous Convalescent Plasma) with a non-immunogenic level of ABO mismatch antibodies which can be used for COVID-19 patients with any type of blood group with desirable simplicity, feasibility, and efficacy.


Asunto(s)
COVID-19/terapia , Técnicas de Inmunoadsorción , Isoanticuerpos/sangre , Plasma , SARS-CoV-2 , Sistema del Grupo Sanguíneo ABO/inmunología , Adsorción , Antígenos de Grupos Sanguíneos , COVID-19/sangre , Frío , Convalecencia , Recuento de Eritrocitos , Eritrocitos/inmunología , Humanos , Inmunización Pasiva/métodos , Isoanticuerpos/inmunología , Sueroterapia para COVID-19
8.
Vox Sang ; 116(1): 106-115, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32770852

RESUMEN

BACKGROUND AND OBJECTIVES: Frequent platelet transfusion may lead to the formation of alloantibodies and immune-mediated platelet destruction. Currently, identifying economic and effective screening methods is necessary for the management of platelet transfusion while different tests were recommended. The present study aims to challenge the performance of slot blotting (SB) and flow cytometry (FC) assays in detecting immune platelet refractoriness. MATERIALS AND METHODS: Sera from 118 patients who received blood components and were clinically suspected of platelet refractoriness were enrolled. Platelet-reactive antibodies were explored in parallel by SB, FC and monoclonal antibody-specific immobilization of platelet antigens (MAIPA) techniques. In a further study, chloroquine-treated platelets were incubated with MAIPA-positive serum, and then, the results of the SB and FC techniques were compared. RESULTS: Using MAIPA as a reference, antibodies were detected in 51 sera, with specificity for human leucocyte antigens (HLA), human platelet antigens (HPA) or both HLA/HPA, in 27, 18 and 6 patients, respectively. The sensitivity and specificity of SB and FC were 86·3%, 88·1%, 82·4% and 95·5%, respectively. The Spearman correlation revealed significant (P < 0·001) correlations between FC (r = 0·763) and SB (r = 0·738) with MAIPA. In respect to HPA antibody detection, SB had 83·3% sensitivity and 92·6% specificity compared to 91·7% and 96·3% for FC while both approaches are acceptable (P < 0·001, r = 0·69; P < 0·001, r = 0·773) and can be recommended. CONCLUSIONS: The present study acknowledges that among the used methods, the flow cytometry's performance is the most appropriate, but slot blotting, with acceptable sensitivity, can be used as an acceptable and convenient procedure for platelet antibody screening.


Asunto(s)
Plaquetas/inmunología , Citometría de Flujo/métodos , Isoanticuerpos/sangre , Trombocitopenia/diagnóstico , Adulto , Antígenos de Plaqueta Humana/inmunología , Femenino , Humanos , Masculino , Transfusión de Plaquetas , Sensibilidad y Especificidad , Trombocitopenia/sangre , Trombocitopenia/inmunología
9.
Cytotechnology ; 72(4): 551-567, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32601794

RESUMEN

As a widely used cell culture supplement, fetal bovine serum (FBS) harbor high content of growth, proliferation, and adhesion factors. However, high cost, bio-safety, possible xenogeneic agent transmission, finite accessible, and ethical controversy are major obstacles that discourage the use of this additive. Accordingly, novel alternatives have been proposed with various pros and cons. Still, caution should be taken in choosing suitable substitute given that the alteration in the main aspects of cultured cells can be biased the consequences of clinical applications. Herein, the authors evaluated the impact of cord blood serum harvesting by hydroxyethyl starch (CBS-HES), as an enriched source of growth factors, on the basic mesenchymal stem cells (MSCs) characteristics. In the present experiment, umbilical cord-derived MSCs were isolated and continuously nourished with Dulbecco's Modified Eagle Medium containing either 10, 15, and 20% CBS-HES or FBSs to compare their morphology, immunophenotype, growth and proliferation rate, death rate, cell cycle, and gene expression profiles. Although all enriched media supported the expansion of MSCs with comparable morphology, cell surface markers, death rate, c-MYC and p16 expression, and growth rate, CBS-HES treated cells significantly (P < 0.05) expressed more hTERT gene in a concentration-dependent manner. Yet no significant shift was observed in the cell cycle of cultured cells using the same concentrations of additives, a finding which further confirmed by Ki-67 immunostaining. CBS-HES as an available and affordable additive, seems to be an optimal, relatively safe, and promising FBS alternative for cultivation, propagation, and subsequent clinical applications of MSCs.

10.
Arch Iran Med ; 23(3): 207-210, 2020 03 01.
Artículo en Inglés | MEDLINE | ID: mdl-32126790

RESUMEN

Hereditary spherocytosis (HS), a familial defect involving red blood cell (RBC) membrane proteins, is associated with reduced deformability, increased fragility, and progressive destruction of spherical cells. The present study focuses on three subjects of a family showing a history of repeated episodes of lethargy and pallor of unknown etiology. All patients displayed reticulocytosis and spherocytosis and one of them had anemia and splenomegaly. The patients underwent screening tests to rule in/out possible underlying disorders, and deficiency/dysfunction of RBC membrane proteins was suspected. Definitive diagnosis can be made on the basis of membrane protein analysis by quantitative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Interestingly, all patients showed marked decrease in the protein 4.2 expression and therefore, HS was confirmed. This case report highlights the simultaneous occurrence of protein 4.2-dependent "typical" and "atypical" HS in a family and serves as a reminder to clinicians to consider RBC membrane disorders in patients presenting with suspicious and unexplained clinical signs.


Asunto(s)
Esferocitosis Hereditaria/sangre , Electroforesis , Membrana Eritrocítica/metabolismo , Familia , Femenino , Humanos , Irán , Masculino , Persona de Mediana Edad , Linaje , Esferocitosis Hereditaria/diagnóstico
11.
BMC Res Notes ; 12(1): 803, 2019 Dec 12.
Artículo en Inglés | MEDLINE | ID: mdl-31831065

RESUMEN

OBJECTIVE: The resistance to antimony-containing glucantime is a major obstacle to successful treatment, especially in endemic areas. Looking the molecular mechanisms involved in this drug resistance will help in choosing the best treatment. The aim of this study was to evaluate the expression of multidrug-resistance 1 (MDR1) and multidrug-resistance protein A (MRPA) genes in acute, chronic non-lupoid, and chronic lupoid forms of dry type cutaneous leishmaniasis (DTCL). RESULTS: MDR1 gene was over-expressed as 14.4- and 1.56-folds in the chronic lupoid and acute forms compared with the chronic non-lupoid form, respectively. Results comparison showed P < 0.05 between the chronic non-lupoid and acute groups, P < 0.01 between acute and chronic lupoid groups, and P < 0.001 between the chronic non-lupoid and chronic lupoid groups. MRPA gene was over-expressed as 266 and 17.7-fold in the chronic lupoid and chronic non-lupoid forms compared with the acute form, respectively. Statistical analysis showed P < 0.01 between the chronic non-lupoid and chronic lupoid groups, P < 0.05 between acute and chronic non-lupoid groups, and P < 0.001 between the acute and chronic lupoid groups.


Asunto(s)
Leishmaniasis Cutánea/metabolismo , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/metabolismo , Subfamilia B de Transportador de Casetes de Unión a ATP/genética , Subfamilia B de Transportador de Casetes de Unión a ATP/metabolismo , Enfermedad Aguda , Enfermedad Crónica , Expresión Génica , Humanos , Leishmaniasis Cutánea/genética , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Piel/metabolismo , Regulación hacia Arriba/genética
12.
BMC Res Notes ; 12(1): 615, 2019 Sep 23.
Artículo en Inglés | MEDLINE | ID: mdl-31547842

RESUMEN

OBJECTIVE: Histopathological studies suggest that parasite load is different between acute and chronic forms of cutaneous leishmaniasis (CL). However, highly sensitive detection methods are still needed to distinguish different forms of leishmaniasis. In the present study, we developed a quantitative real-time polymerase chain reaction (PCR) to detect and quantify Leishmania tropica parasites in paraffin-embedded tissue samples. RESULTS: The ability of real-time PCR for leishmania detection was higher than histopathological evaluation. The quantitative real-time PCR (qPCR) quantified parasite loads were highly correlated with microscopic results (r = 0.598; P < 0.001). Among patients, the parasite load was inversely correlated with disease duration (acute CL lesions had very higher parasite load than chronic CL lesions), but there was no difference in the parasite load according to the patients' age and sex as well as location of the lesions. In contrast to Ridley scoring system (P < 0.001), there were no statistically significant differences in the relative number of parasites among the lupoid and non-lupoid forms of chronic lesions in real-time PCR (P = 0.549), which indicates the superiority of histopathological evaluation for chronic forms differentiation.


Asunto(s)
ADN Protozoario/genética , Leishmania tropica/genética , Leishmaniasis Cutánea/diagnóstico , Piel/parasitología , Enfermedad Aguda , Adolescente , Adulto , Anciano , Niño , Enfermedad Crónica , ADN Protozoario/clasificación , Diagnóstico Diferencial , Femenino , Histocitoquímica , Humanos , Leishmania tropica/clasificación , Leishmaniasis Cutánea/clasificación , Leishmaniasis Cutánea/parasitología , Leishmaniasis Cutánea/patología , Masculino , Persona de Mediana Edad , Adhesión en Parafina , Carga de Parásitos , Reacción en Cadena en Tiempo Real de la Polimerasa , Piel/patología
13.
Biomater Sci ; 7(5): 1949-1961, 2019 Apr 23.
Artículo en Inglés | MEDLINE | ID: mdl-30793722

RESUMEN

The application of conventional approaches to diabetic wound regeneration has some limitations. Thus, skin substitutes could be a new therapeutic possibility. In this regard, fibrin scaffolds are promising materials due to their desirable characteristics. Since defective fibroblasts caused by diabetes can disrupt regeneration, it seems that the use of living cells can improve the healing process. Thus, based on this fact, a cellular fibrin membrane was used to evaluate the diabetic wound healing in rats. The fibrin membrane was fabricated using fresh frozen plasma on which isolated fibroblasts were cultured. The wound model was created on 36 diabetic rats that were randomly divided into three groups: control, membrane, and cellular fibrin membrane (CM). Wound photogramography and immuno-histopathological staining were performed during consecutive days after treatment. Macroscopic evaluation of the wounds indicated a noteworthy enhancement of wound closure in the CM group. In the CM group, the re-epithelialization rate on day 7, 10 (p < 0.001), and 14 (p < 0.05), the fibroblast percentage on day 3 (p < 0.01) and 7 (p < 0.05) and the collagenization in all days were significantly higher than those of other groups (p < 0.001). The fibroblast number in the CM group on day 10 was significantly (p < 0.01) lower than that in the other groups. Contrary to the neutrophil and angiogenesis percentages that had no significant difference among the groups at different points of time (p > 0.05), the macrophage percentage on day 7 (P < 0.01), 10, and 14 (p < 0.05) was significantly lower in the CM group as compared to that in other groups. Overall, it seems that the use of a fibroblast-loaded fibrin membrane is an attractive strategy to promote diabetic wound healing.


Asunto(s)
Células Alogénicas/citología , Materiales Biocompatibles/farmacología , Diabetes Mellitus Experimental/fisiopatología , Fibroblastos/citología , Membranas Artificiales , Cicatrización de Heridas/efectos de los fármacos , Animales , Materiales Biocompatibles/metabolismo , Recuento de Células , Colágeno/metabolismo , Epitelio/efectos de los fármacos , Epitelio/metabolismo , Fibrina/metabolismo , Macrófagos/citología , Macrófagos/efectos de los fármacos , Masculino , Neovascularización Fisiológica/efectos de los fármacos , Neutrófilos/citología , Neutrófilos/efectos de los fármacos , Ratas , Ratas Wistar , Piel/irrigación sanguínea , Piel/efectos de los fármacos , Piel/inmunología , Piel/patología
14.
Arch Iran Med ; 21(8): 362-367, 2018 08 01.
Artículo en Inglés | MEDLINE | ID: mdl-30113858

RESUMEN

BACKGROUND: The mesenchymal stem cells (MSCs) of peripheral blood (PB) have been recognized as a promising source for allogeneic cell therapy. The objective of the present study was to isolate and characterize MSCs derived from non-mobilized PB, and evaluate their differentiation potential. METHODS: The buffy coat mononuclear fractions of the PB were concentrated using the Ficoll-Paque density gradient centrifugation and were grown on primary and secondary culture media, respectively. The isolated cells were characterized using a multidisciplinary approach which was based on morphology, immunophenotyping, gene expression, and multipotentiality. Flow cytometry and Reverse transcription polymerase chain reaction (RT-PCR) were used to identify the expression of different MSC markers. Finally, after culturing in osteogenic and adipogenic induction media, the isolated cells were stained by Alizarin red and Oil-Red O. RESULTS: In spite of absence of any bone marrow stimulating factor, the isolation approach in this study yielded a rather homogeneous and spindle-shaped mononuclear cell population (the yield of passage 0 was 0.65 ± 0.15) that stained positive for CD90, CD105, and CD73, and were negative for CD45 and CD34. These cells have high proliferative capacity (confirmed by the expression of Oct-4, Nucleostemin, and Nanog genes) and were able to differentiate into lineage-specific committed cells, when exposed to the appropriate medium. CONCLUSION: Overall, it can be concluded that conventional, labour-intensive and time-consuming approaches are not necessary in isolating MSCs from PB. This relatively accessible and minimally invasive source, PB, represents a good alternative reservoir of homogeneous MSCs that could open a new era for practical exploitation in regenerative medicine.


Asunto(s)
Células de la Médula Ósea/citología , Diferenciación Celular/fisiología , Separación Celular/métodos , Células Madre Mesenquimatosas/citología , Proliferación Celular , Células Cultivadas , Citometría de Flujo , Humanos , Inmunohistoquímica , Inmunofenotipificación , Reacción en Cadena de la Polimerasa
15.
Iran J Pathol ; 13(4): 461-466, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30774686

RESUMEN

BACKGROUND AND OBJECTIVES: The mesenchymal stem cells derived from peripheral blood (PB) have been recognized as a promising source for allogeneic cell therapy. The aim of this study was to investigate the isolation, growth and differentiation ability of peripheral blood-isolated mesenchymal stem cells. METHODS: The mononuclear cells were purified from fresh peripheral blood using density gradient centrifugation then cultured in a suitable medium, expanded and characterized. In the following, these cells were cultured in specific adipogenic and osteogenic differentiation media. RESULTS AND CONCLUSION: In spite of the absence of any stimulating factor, the cells adhered to the flasks and developed a rather homogeneous, spindle-shaped morphology after consecutive passages. The cells were confirmed to have mesenchymal phenotype by expression of specific markers (CD90, CD105, and CD73) and absence of CD45 marker, which is specific for hematopoietic stem cells. They could differentiate into lineage-specific committed cells (osteoblasts and adipocytes).According to the findings, the conventional, labour-intensive and time-consuming approaches are not necessary to obtain an optimal number of cells from peripheral blood. This relatively accessible and minimally invasive source of stem cells may open a new era for practical exploitation in regenerative medicine.

16.
Blood Coagul Fibrinolysis ; 28(8): 681-686, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29084015

RESUMEN

: Quantitative and/or qualitative defects of the platelet membrane glycoprotein IIb/IIIa complex lead to the clinical entity of Glanzmann's thrombasthenia. A large variety of mutations and polymorphisms are responsible for the aberrant expression and defective activity of this heterodimeric complex. The current study aimed to determine the pattern of mutations among Iranian population with Glanzmann's thrombasthenia. A total of 20 patients with Glanzmann's thrombasthenia have been evaluated. All exons and splice sites of ITGA2B and ITGB3 genes were amplified using touchdown PCR. Mutation screening was analyzed using conformation sensitive gel electrophoresis heteroduplex PCR, and DNA sequencing. In addition to finding one previously identified mutation and polymorphism, the experimenters explored 3 and 2 novel mutations and polymorphisms, respectively. One substitution mutation, two deletions of a single nucleotide, one insertion of a single nucleotide, two synonymous polymorphisms, and one missense polymorphism were found using Sanger sequencing method. All detected mutations were homozygous which will most likely contribute to the pathogenesis of Glanzmann's thrombasthenia. Furthermore, it suggested ITGB3 as the mainly affected gene impaired in the patients with Glanzmann's thrombasthenia. As expected, the molecular results were consistent with the phenotypic findings so that GPIIb/IIIa complex was disrupted due to mutations in all type-I Glanzmann's thrombasthenia patients. It is concluded that intronic alterations or epigenetic regulations could be responsible for aberrant expression and/or defective activity of GPIIb/IIIa complex among other patients.


Asunto(s)
Mutación , Trombastenia/genética , Homocigoto , Humanos , Integrina alfa2 , Integrina beta3 , Irán/epidemiología , Epidemiología Molecular , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/genética , Análisis de Secuencia de ADN
18.
Neurotox Res ; 32(1): 41-49, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28144902

RESUMEN

Despite the importance of this issue, less has been paid to the influence of exercise on the neural side effects of anabolic androgenic steroids and mechanisms. We investigated the effects of two levels of endurance exercise on neurodegeneration side effects of nandrolone. The study period was 8 weeks. Wistar rats were divided into nine groups including the control (CTL) group, mild exercise (mEx) group, and vehicle (Arach) group which received arachis oil intramuscularly, nandrolone (Nan) group which received nandrolone decanoate 5 mg/kg two times weekly, mEx+Arach group which treated with arachis oil along with mild exercise, mEx+Nan group which treated with nandrolone along with mild exercise, severe exercise (sEx) group, sEx+Arach, and sEx+Nan groups. Finally, brain samples were taken for histopathological, biochemical, and western blot analysis. Nandrolone significantly decreased the intact cells of the hippocampus, total antioxidant capacity (TAC) (P < 0.05 versus CTL and Arach groups), TAC to malondialdehyde ratio (TAC/MDA), and Bcl-2. Nandrolone increased the Bax/Bcl-2 ratio of the brain tissue (P < 0.01 versus CTL and Arach groups). Combination of mild exercise and nandrolone rescued the intact cells to some extent, and this effect was associated with the improvement of Bcl-2 level and Bax/Bcl-2 ratio of brain tissue. Combination of severe exercise and nandrolone rescued the intact cells and improved the TAC, TAC/MDA, and Bax/Bcl-2 ratios. The findings suggest that low- and high-intensity endurance exercise decreased the risk of neurodegeneration effect of nandrolone in the hippocampus of rats. This effect can be explained by the regulation of the redox system and cell homeostasis.


Asunto(s)
Anabolizantes/toxicidad , Apoptosis/efectos de los fármacos , Nandrolona/análogos & derivados , Enfermedades Neurodegenerativas/inducido químicamente , Enfermedades Neurodegenerativas/rehabilitación , Oxidación-Reducción/efectos de los fármacos , Condicionamiento Físico Animal/fisiología , Animales , Modelos Animales de Enfermedad , Masculino , Malondialdehído/metabolismo , Nandrolona/toxicidad , Nandrolona Decanoato , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Ratas Wistar , Natación , Proteína X Asociada a bcl-2/metabolismo
20.
Sci Rep ; 6: 31866, 2016 08 18.
Artículo en Inglés | MEDLINE | ID: mdl-27535591

RESUMEN

Traumatic brain injury (TBI) is a major health concern affecting the general public as well as military personnel. However, there is no FDA-approved therapy for the treatment of TBIs. In this work, we investigated the neurotherapeutic effects of the well-known natural Iranian medicine Satureja Khuzistanica Jamzad (SKJ) essential oil (SKEO) on the outcomes of diffused experimental TBI, with particular attention paid to its anti-inflammatory and anti-apoptotic effects. Male Wistar rats were treated with doses of 50, 100 and 200 (mg/kg, i.p) SKEO after induction of diffused TBIs. The results showed that injecting SKEO (200 mg/kg) 30 minutes after TBI significantly reduced brain oedema and damage to the blood-brain barrier (BBB) and limited the post-TBI increase in intracranial pressure. The veterinary coma scale (VCS) scores significantly improved in the treatment group. Also, inflammatory marker assays showed reduced levels of TNF-α, IL-1ß, and IL-6 and increased IL-10 in the treated groups. Moreover, the immunohistochemical results indicated that SKEO not only reduced neuronal death and BBB permeability but also affected astrocytic activation. Overall, our data indicate potential clinical neurological applications for SKEO.


Asunto(s)
Antiinflamatorios/farmacología , Lesiones Traumáticas del Encéfalo/tratamiento farmacológico , Aceites Volátiles/farmacología , Satureja/química , Animales , Antiinflamatorios/química , Barrera Hematoencefálica/metabolismo , Barrera Hematoencefálica/patología , Lesiones Traumáticas del Encéfalo/metabolismo , Lesiones Traumáticas del Encéfalo/patología , Citocinas/metabolismo , Presión Intracraneal/efectos de los fármacos , Masculino , Aceites Volátiles/química , Ratas , Ratas Wistar
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