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1.
Braz. j. biol ; 842024.
Article in English | LILACS-Express | LILACS, VETINDEX | ID: biblio-1469262

ABSTRACT

Abstract Mammals have a limited capacity to regenerate their tissues and organs. One of the mechanisms associated with natural regeneration is dedifferentiation. Several small molecules such as vitamin C and growth factors could improve reprogramming efficiency. In this study, the NTERA2-D1 (NT2) cells were induced towards differentiation (NT2-RA) with 10-5 M retinoic acid (RA) for three days and then subjected to various amounts of vitreous humor (VH). Results show that the growth rate of these cells was reduced, while this rate was partly restored upon treatment with VH (NT2-RA-VH). Cell cycle analysis with PI method also showed that the numbers of cells at the S phase of the cell cycle in these cells were increased. The levels of SSEA3 and TRA-1-81 antigens in NT2-RA were dropped but they increased in NT2- RA-VH to a level similar to the NT2 cells. The level of SSEA1 had an opposite pattern. Expression of OCT4 gene dropped after RA treatment, but it was recovered in NT2-RA-VH cells. In conclusion, we suggest VH as a potent mixture for improving the cellular reprogramming leading to dedifferentiation.


Resumo Os mamíferos têm uma capacidade limitada de regenerar seus tecidos e órgãos. Um dos mecanismos associados à regeneração natural é a desdiferenciação. Várias moléculas pequenas, como vitamina C e fatores de crescimento, podem melhorar a eficiência da reprogramação. Neste estudo, as células NTERA2-D1 (NT2) foram induzidas à diferenciação (NT2-RA) com ácido retinóico (RA) 10-5 M por três dias e depois submetidas a várias quantidades de humor vítreo (VH). Os resultados mostram que a taxa de crescimento dessas células foi reduzida, enquanto essa taxa foi parcialmente restaurada após o tratamento com VH (NT2-RA-VH). A análise do ciclo celular com o método PI também mostrou que o número de células na fase S do ciclo celular nessas células estava aumentado. Os níveis de antígenos SSEA3 e TRA-1-81 em NT2-RA diminuíram, mas aumentaram em NT2-RA-VH a um nível semelhante ao das células NT2. O nível de SSEA1 teve um padrão oposto. A expressão do gene OCT4 diminuiu após o tratamento com AR, mas foi recuperado em células NT2-RA-VH. Em conclusão, sugerimos o VH como uma mistura potente para melhorar a reprogramação celular levando à desdiferenciação.

2.
Braz. j. biol ; 84: e250151, 2024. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1350306

ABSTRACT

Abstract Mammals have a limited capacity to regenerate their tissues and organs. One of the mechanisms associated with natural regeneration is dedifferentiation. Several small molecules such as vitamin C and growth factors could improve reprogramming efficiency. In this study, the NTERA2-D1 (NT2) cells were induced towards differentiation (NT2-RA) with 10-5 M retinoic acid (RA) for three days and then subjected to various amounts of vitreous humor (VH). Results show that the growth rate of these cells was reduced, while this rate was partly restored upon treatment with VH (NT2-RA-VH). Cell cycle analysis with PI method also showed that the numbers of cells at the S phase of the cell cycle in these cells were increased. The levels of SSEA3 and TRA-1-81 antigens in NT2-RA were dropped but they increased in NT2- RA-VH to a level similar to the NT2 cells. The level of SSEA1 had an opposite pattern. Expression of OCT4 gene dropped after RA treatment, but it was recovered in NT2-RA-VH cells. In conclusion, we suggest VH as a potent mixture for improving the cellular reprogramming leading to dedifferentiation.


Resumo Os mamíferos têm uma capacidade limitada de regenerar seus tecidos e órgãos. Um dos mecanismos associados à regeneração natural é a desdiferenciação. Várias moléculas pequenas, como vitamina C e fatores de crescimento, podem melhorar a eficiência da reprogramação. Neste estudo, as células NTERA2-D1 (NT2) foram induzidas à diferenciação (NT2-RA) com ácido retinóico (RA) 10-5 M por três dias e depois submetidas a várias quantidades de humor vítreo (VH). Os resultados mostram que a taxa de crescimento dessas células foi reduzida, enquanto essa taxa foi parcialmente restaurada após o tratamento com VH (NT2-RA-VH). A análise do ciclo celular com o método PI também mostrou que o número de células na fase S do ciclo celular nessas células estava aumentado. Os níveis de antígenos SSEA3 e TRA-1-81 em NT2-RA diminuíram, mas aumentaram em NT2-RA-VH a um nível semelhante ao das células NT2. O nível de SSEA1 teve um padrão oposto. A expressão do gene OCT4 diminuiu após o tratamento com AR, mas foi recuperado em células NT2-RA-VH. Em conclusão, sugerimos o VH como uma mistura potente para melhorar a reprogramação celular levando à desdiferenciação.


Subject(s)
Humans , Vitreous Body , Cell Proliferation , Cell Dedifferentiation , Tretinoin , Tumor Cells, Cultured , Cell Differentiation , Cell Division , Cell Line
3.
Braz J Biol ; 84: e250151, 2021.
Article in English | MEDLINE | ID: mdl-34817042

ABSTRACT

Mammals have a limited capacity to regenerate their tissues and organs. One of the mechanisms associated with natural regeneration is dedifferentiation. Several small molecules such as vitamin C and growth factors could improve reprogramming efficiency. In this study, the NTERA2-D1 (NT2) cells were induced towards differentiation (NT2-RA) with 10-5 M retinoic acid (RA) for three days and then subjected to various amounts of vitreous humor (VH). Results show that the growth rate of these cells was reduced, while this rate was partly restored upon treatment with VH (NT2-RA-VH). Cell cycle analysis with PI method also showed that the numbers of cells at the S phase of the cell cycle in these cells were increased. The levels of SSEA3 and TRA-1-81 antigens in NT2-RA were dropped but they increased in NT2- RA-VH to a level similar to the NT2 cells. The level of SSEA1 had an opposite pattern. Expression of OCT4 gene dropped after RA treatment, but it was recovered in NT2-RA-VH cells. In conclusion, we suggest VH as a potent mixture for improving the cellular reprogramming leading to dedifferentiation.


Subject(s)
Cell Dedifferentiation , Cell Proliferation , Vitreous Body , Cell Differentiation , Cell Division , Cell Line , Humans , Tretinoin , Tumor Cells, Cultured
4.
Allergol. immunopatol ; 48(6): 711-719, nov.-dic. 2020. tab
Article in English | IBECS | ID: ibc-199262

ABSTRACT

BACKGROUND: Common Variable Immunodeficiency (CVID) is characterized by an impaired antibody production and a higher susceptibility to encapsulated bacterial infections. Lung disease is considered to be the most important cause of morbidity and mortality. METHODS: We analyzed clinical, radiological and functional characteristics in 80 patients with CVID assisted in the Unidad Inmunologia e Histocompatibilidad at Durand Hospital from 1982 to 2018. RESULTS: Of the 80 patients, 55 showed pathologic lung Computed Tomography (CT). Twenty of them (36.4%) showed bronchiectasis; 26 (47.3%) interstitial involvement associated with nodules and adenopathies called GLILD (granulomatous-lymphocytic interstitial lung disease); and nine patients (16.3%) showed other lesions. Nine percent of patients with lung disease showed CT progression; none of them had spirometry worsening. GLILD patients had normal and restrictive patterns in lung function tests, in equal proportions. Two patients - one with GLILD and the other one with bronchiectasis - had an increase in spirometric pattern severity without CT progression. Lung biopsy was performed in 19% of GLILD patients, all of whom had histopathologic diagnosis of Lymphoid Interstitial Pneumonia (LIP). CONCLUSIONS: GLILD is the major cause of lung disease in CVID. Computed tomography is useful for diagnosis but not necessary in follow-up, in which functional tests should have better correlation with clinical evolution, reducing radiation exposure. Biopsy should be indicated when the clinical diagnosis is unclear. Treatment should be considered whenever there is clear evidence of disease progression


No disponible


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Immunologic Deficiency Syndromes/diagnosis , Delayed Diagnosis/statistics & numerical data , Immunologic Deficiency Syndromes/pathology , Immunologic Deficiency Syndromes/epidemiology , Severity of Illness Index , Risk Factors , Cross-Sectional Studies , Statistics, Nonparametric , Logistic Models , Iran/epidemiology , Prevalence
5.
Allergol Immunopathol (Madr) ; 48(6): 711-719, 2020.
Article in English | MEDLINE | ID: mdl-32404246

ABSTRACT

INTRODUCTION: The prevalence of undiagnosed primary immunodeficiency diseases is remarkably high and contributes to increasing the rate of morbidity and mortality among this group of patients. OBJECTIVE: To examine the 10 warning sign scoring system in patients suspected of primary immune deficiency and also estimate the diagnostic delay in patients with proven disease. METHODS: This descriptive cross-sectional study was carried out during the years 2015-2016 in Ali Asghar (AS) Clinic and Hospital. Two hundred patients with suspected primary immune deficiency disease were eligible for inclusion in the study. Multivariable logistic regression analysis was used to determine the relation between findings. RESULTS: In this study, the majority of suspected cases of immunodeficiency were males (57%) with a mean age of 3.33±2.89 years. Twenty-one (10.5%) patients were diagnosed with immunodeficiency disease. The mean diagnostic delay among primary immune deficient patients was 2.05±1.7 years. There was a significant relationship between having parental consanguinity (OR=2.68, 95% CI: 1.07-6.70), allergies (OR=5.03, 95% CI: 1.13-22.31), vaccine adverse effects (OR=9.31, 95% CI: 1.24-69.96) and primary immune deficiency diagnosis. No association was observed between age (OR=0.98, 95% CI: 0.84-1.14), gender (OR=0.99, 95% CI: 0.39-2.47), immune deficiency scoring (OR=0.68, 95% CI: 0.31-1.45) and primary immune deficiency diagnosis. CONCLUSION: Ten warning sign scoring system is of less value to consider a patient suspected of having primary immune deficiency. There is a meaningful delay in diagnosis of primary immune deficiencies especially in antibody deficiency defects which seeks further upgrading of knowledge in physicians.


Subject(s)
Consanguinity , Delayed Diagnosis/statistics & numerical data , Primary Immunodeficiency Diseases/epidemiology , Vaccines/adverse effects , Child , Child, Preschool , Cross-Sectional Studies , Female , Humans , Infant , Iran/epidemiology , Logistic Models , Male , Multivariate Analysis , Prevalence , Primary Immunodeficiency Diseases/diagnosis , Primary Immunodeficiency Diseases/immunology , Risk Factors
6.
Arch Razi Inst ; 74(1): 1-6, 2019 03.
Article in English | MEDLINE | ID: mdl-31013002

ABSTRACT

Brucella bacterium causes Brucellosis, an infectious disease spreading from animals to human. Brucella lumazine synthase (BLS) is a highly immunogenic protein with adjuvant properties, which has been introduced as an effective protein carrier for vaccine development. This protein also plays a significant role in inducing immune system. This study aimed to clone, express, and purify the BLS gene from Brucella melitensis Rev1. The BLS gene was amplified by particular primers with the restriction enzyme sites as a linker and it was inserted into pTZ57R/T vector. Subsequently, it was ligated into pET32(a)+ expression vector. Recombinant expression vector containing coding sequence of BLS was transformed into E. coli BL21 (DE3) host gene expression and stimulated by 0.1mM IPTG. The results of sequencing showed that there were not any mutations in BLS encoding sequence. The expression results were set by sequencing and endorsed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses and western blotting that showed 35 kDa protein band appropriately.


Subject(s)
Bacterial Proteins/immunology , Brucella Vaccine/immunology , Brucella melitensis/immunology , Brucellosis/veterinary , Genes, Bacterial , Multienzyme Complexes/immunology , Blotting, Western/veterinary , Brucellosis/prevention & control , Electrophoresis, Polyacrylamide Gel/veterinary , Escherichia coli/genetics , Microorganisms, Genetically-Modified/genetics , Recombinant Proteins/immunology , Vaccines, Subunit/immunology
7.
Rev Sci Instrum ; 90(12): 125107, 2019 Dec 01.
Article in English | MEDLINE | ID: mdl-31893769

ABSTRACT

An experimental flow control technique is given in this paper to study the jet effect on the coflow jet's airfoil with injection and suction and compared with the jet-off condition. The airfoil is CFJ0025-065-196, and the Reynolds number based on the airfoil's chord length is 105. To measure the turbulence components of flow, a hot wire anemometry apparatus in a wind tunnel has been used. In this paper, the effect of the average velocity and boundary layer thickness on the coflow jet's airfoil is analyzed. The test is done for two different coflow velocities and for different angles of attack. It is also shown that, by increasing the velocity difference between the jet and the main flow, separation is delayed, and this delay can be preserved by raising coflow velocity at higher angles of attack. So, this flow control method has a good efficiency, and it is possible to reach higher numbers of lift and lower numbers of drag coefficients.

8.
Gene Ther ; 25(2): 93-103, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29523878

ABSTRACT

The use of immunomodulatory cytokines has been shown effective in regressing a wide range of tumors. However, systemic delivery of recombinant cytokines results in serious, potentially life-threatening, adverse effects. By contrast, nucleic acid transfer via electroporation (EP) is a safe and effective method of delivering plasmid-encoded cytokines to tumors. Intratumoral delivery of IL-12 plasmid DNA by electroporation (IT-pIL12-EP) produced objective response rates in Phase 2 clinical trials in metastatic melanoma. However, only 17.9% of patients receiving IT-pIL12-EP show a complete therapeutic response. Here, we sought to improve the antitumor efficacy of our clinical IT-pIL12-EP plasmid electroporation platform. We evaluated multiple plasmid designs for IL-12 expression. IL-12 expression from a plasmid incorporating a picornavirus-derived co-translational P2A site was the most effective in expressing IL-12p70. In addition, modifying the electroporation parameters improved transfection efficiency and expression of plasmid-derived IL-12p70, as well as its downstream effector IFN-γ in vivo. Finally, using a murine melanoma model that is representative of the intended target patient population, we show that combining modified electroporation conditions with the pIL12-P2A plasmid expression enhances the systemic antitumor response. These improvements to the IT-pIL12-EP platform may improve patient clinical response rates and survival when translated to clinical trials.


Subject(s)
Electroporation/methods , Gene Transfer Techniques , Interleukin-12/genetics , Melanoma, Experimental/therapy , Plasmids , Animals , CD4-CD8 Ratio , Enzyme-Linked Immunosorbent Assay , HEK293 Cells , Humans , Injections, Intralesional , Interferon-gamma/blood , Interferon-gamma/metabolism , Interleukin-12/biosynthesis , Internal Ribosome Entry Sites , Melanoma, Experimental/immunology , Mice , Picornaviridae/genetics
9.
Cell Mol Biol (Noisy-le-grand) ; 62(10): 26-31, 2016 Aug 31.
Article in English | MEDLINE | ID: mdl-27609470

ABSTRACT

Variegation in flower color is commonly observed in many plant species and also occurs on Petunia (Petunia hybrida) as an ornamental plant. Variegated plants are of highly valuable in the floricultural market. Agroinfiltration is an Agrobacterium-mediated transient assay for the analysis of gene function and genetic modification in leaves, flowers and fruit tissues of various plants. Transient RNAi-induced silencing by agroinfiltration has been developed in leaves and fruits of several plant species. Here we report the establishment of a transient hairpin RNAi-induced silencing system for color modification assay in floral tissues of Petunia with different colors. chiRNAi construct was cloned into the pBI121 vector under the control of 35S promoter. Transient RNA silencing of chi in the floral tissues of Petunia was induced by delivering 530 bp chi hairpin RNAs (hpRNAs) into the petals of flowers using agroinfiltration. Impaired anthocyanin accumulation and reduction of endogenous mRNAs of the corresponding targets were observed in the infiltrated areas of the petals of four colors of Petunia. Silencing of the endogenous chi mRNAs was highly effective in reduction of chi gene and anthocyanin accumulation. This transient silencing system is a prototype for modification of the anthocyanin biosynthetic pathway in Petunia through chi gene suppression.


Subject(s)
Agrobacterium/metabolism , Flowers/enzymology , Flowers/genetics , Genetic Techniques , Intramolecular Lyases/genetics , RNA Interference , Base Pairing/genetics , Gene Expression Regulation, Plant , Phenotype , Pigmentation , Real-Time Polymerase Chain Reaction
10.
Cell Mol Biol (Noisy-le-grand) ; 62(10): 68-74, 2016 Aug 31.
Article in English | MEDLINE | ID: mdl-27609477

ABSTRACT

Mesenchymal stem cells (MSCs) have been considered promising tools for new clinical concepts in supporting cellular therapy and regenerative medicine. More recently, Ream/Irrigator/Aspirator (RIA) was introduced as a source of MSCs. In this study we compared MSCs derived from three different sources (iliac crest bone marrow (ICBM), adipose tissue (AT), and (RIA)) regarding the morphology, the success rate of isolating MSCs, colony frequency, expansion potential, osteogenic and chondrogenic differentiation capacity. MSCs were isolated from three different sources and flow cytometric analyses were performed for cell characterization. Colony-forming unit-fibroblast (CFU-F) assay and population doubling time (PDT) were evaluated for MSCs derived from three different sources and differentiation potential of RIA, ICBM-, and AT-MSCs were determined by staining. Additionally, gene expression profiles for tissue specific markers corresponding to osteogenesis and chondrogenesis were analyzed using real time polymerase chain reaction (RT-PCR). Cultured with the appropriate condition, osteogenic and chondrogenic differentiation could be confirmed in all MSC preparations. Flow cytometry analysis indicated that RIA- and AT-derived MSCs have more homogenous populations than ICBM-MSCs. A comparison of the colonogenic ability in different tissues by CFU-F assay after 10 days showed that more colonies are formed from RIA-MSCs than from ICBM-MSCs, and AT-MSCs. AT-MSCs, were dispersed with no obvious colonies. The RIA-MSCs underwent osteogenesis and chondrogenesis at a faster rate than ICBM and AT-MSCs. Direct comparisons of RIA- to ICBM- and AT-MSCs have shown the RIA-MSCs have higher differentiation toward osteoblast and chondrocytes compared to other sources of MSCs. Hence, RIA-MSCs may be recommended as a more suitable source for treating orthopedic disorders.


Subject(s)
Adipose Tissue/cytology , Bone Marrow Cells/cytology , Ilium/cytology , Mesenchymal Stem Cells/cytology , Suction/instrumentation , Biomarkers/metabolism , Cell Differentiation , Cell Lineage , Cell Membrane/metabolism , Cellular Senescence , Chondrocytes/cytology , Chondrogenesis , Clone Cells , Colony-Forming Units Assay , Humans , Osteoblasts/cytology , Osteogenesis
11.
Allergol. immunopatol ; 44(4): 322-330, jul.-ago. 2016. graf, tab
Article in English | IBECS | ID: ibc-154434

ABSTRACT

PURPOSE: The aim was to describe the clinical manifestations, complications and long-term outcome of a cohort of Iranian patients with primary immune deficiency (PID). METHOD: We retrospectively studied the demographic, clinical and immunological characteristics of the PID patients in a single tertiary centre, from January 1989 to July 2014. The patients were classified according to the International Union of Immunological Societies Expert Committee on PID. RESULTS: 98 patients were diagnosed with and followed-up for 15 disorders. The mean age at onset and diagnosis and the diagnostic delay were 8±10, 14.2±13.1 and 6.1±7 years, respectively. Parental consanguinity rate was 57%. Predominantly Antibody Deficiency was the most common diagnosis (n=63), followed by congenital defects of phagocytes (n=16), combined immunodeficiencies (n=12), well defined syndromes (n=4) and defects in innate immunity (n=3). Recurrent sinopulmonary infection was the most common presentation. Active infections were treated appropriately, in addition to prophylactic therapy with IVIG and antimicrobials. Not all the patients were compliant with prophylactic regimens due to cost and unavailability. One SCID patient underwent successful bone marrow transplantation. The total mortality rate was 19% during the follow-up period (7.8±7.6 years). The mean age of living patients at the time of study was 23±11.7 years. CONCLUSIONS: Physicians awareness of PID has been rising dramatically in Iran, ensuring an increasing number of patients being diagnosed and treated. More effective treatment services, including health insurance coverage and drug availability are needed to improve the outcome of PID patients


No disponible


Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Immunologic Deficiency Syndromes/diagnosis , Immunologic Deficiency Syndromes/immunology , Immunologic Deficiency Syndromes/pathology , Immunologic Surveillance , Immunologic Surveillance/immunology , Monitoring, Immunologic/instrumentation , Monitoring, Immunologic/methods , Desensitization, Immunologic , Laboratory Test/methods , Immunoglobulin G/analysis , Immunoglobulin G/immunology , Cohort Studies , Immunocompetence/immunology
12.
Cell Mol Biol (Noisy-le-grand) ; 62(5): 31-7, 2016 Apr 30.
Article in English | MEDLINE | ID: mdl-27188867

ABSTRACT

Targeted homing of transplanted mesenchymal stem cells (MSCs) is a decades old discussion in regenerative medicine. It has been proved that stromal cell-derived factor-1 (SDF-1α) is a potent chemoattractant of MSCs. Therefore, different strategies have been used to increase secretion of SDF-1α in damaged tissues to elevate targeted homing of MSCs. Previous studies have revealed that increased SDF-1α expression in hypoxic necrotic tissues and also low-level laser exposure enhanced angiogenesis in injured tissues. Herein, human skeletal and cardiac muscle cells (HSKM and HCM) were treated with hypoxia and low level laser to see their effects on expression of SDF-1α and on MSCs migration towards these treated cells. The optimal treatment conditions were determined by investigating the cellular viability after treatment. Real-Time PCR and Western blot analysis were done to study the expression of SDF-1α in treated cells. Migration potential of MSCs toward hypoxic and laser treated cells was investigated via migration assay. MTT assay revealed that laser and hypoxia treatment had no effect on the viability of HCM, HSKM compared with Glioblastoma cells. Real-Time PCR showed 16- and 90-fold elevation in mRNA of SDF-1α in HSKM and HCM cells, respectively, in laser treated with 12 J/cm2 intensity. In these two groups, selected as optimal conditions, HIF-1α expression showed maximum fold changes that might be partly because of response to treatments help to SDF-1α expression. It can be concluded that hypoxia and laser treatments may recruit MSCs and applied as a useful strategy for the further targeted stem cell homing.


Subject(s)
Chemokine CXCL12/genetics , Lasers , Muscle Cells/metabolism , Cell Count , Cell Hypoxia/genetics , Cell Movement , Cell Proliferation , Cell Shape , Cells, Cultured , Chemokine CXCL12/metabolism , Gene Expression Regulation , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Myocytes, Cardiac/metabolism , Protein Biosynthesis/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Real-Time Polymerase Chain Reaction , Transcription, Genetic
13.
Cell Mol Biol (Noisy-le-grand) ; 62(5): 44-54, 2016 Apr 30.
Article in English | MEDLINE | ID: mdl-27188869

ABSTRACT

Cutaneous wound healing is a complex type of biological event involving proliferation, differentiation, reprograming, trans/de-differentiation, recruitment, migration, and apoptosis of a number of cells (keratinocytes, fibroblasts, endothelial cells, nerve cells and stem cells) to regenerate a multi-layered tissue that is damaged by either internal or external factors. The exact regeneration mechanism of damaged skin is still unknown but the epithelial and other kinds of stem cells located in skin play crucial roles in the healing process. In this work, a co-culture model composed of adipose derived mesenchymal stem cells and keratinocytes was developed to understand the cellular differentiation behaviour in wound healing. Human mesenchymal stem cells were isolated from waste lipoaspirates. Keratinocytes were isolated from neonatal rats skin as well from human adult skin. Both types of cells were cultured and their culturing behaviour was observed microscopically under regular intervals of time. The identity of both cells was confirmed by flow cytometry and qRT-PCR. Cells were co-cultured under the proposed co-culturing model and the model was observed for 7, 14 and 21 days. The cellular behaviour was studied based on change in morphology, colonization, stratification, migration and expression of molecular markers. Expression of molecular markers was studied at transcriptional level and change in cellular morphology and migration capabilities was observed under the invert microscope regularly. Successfully isolated and characterized mesenchymal stem cells were found to express keratinocyte lineage markers i.e. K5, K10, K14, K18, K19 and Involucrin when co-cultured with keratinocytes after 14 and 21 days. Their expression was found to increase by increasing the time span of cell culturing. The keratinocyte colonies started to disappear after 10 days of culturing which might be due to stratification process initiated by possibly transdifferentiated stem cells. It can be concluded that mesenchymal stem cells can regenerate the damaged skin if transplanted to damaged area but for their successful differentiation and enhanced regeneration, they need a population of keratinocytes in situ which need further experiments for validation of co-culture model and its potential for being used in clinics.


Subject(s)
Adipose Tissue/cytology , Biomarkers/metabolism , Cell Lineage , Coculture Techniques/methods , Keratinocytes/cytology , Mesenchymal Stem Cells/cytology , Adipocytes/cytology , Animals , Animals, Newborn , Cell Differentiation , Cell Proliferation , Cell Separation , Cells, Cultured , Flow Cytometry , Humans , Mesenchymal Stem Cell Transplantation , Osteoblasts/cytology , Rats , Wound Healing
14.
Allergol Immunopathol (Madr) ; 44(4): 322-30, 2016.
Article in English | MEDLINE | ID: mdl-26803694

ABSTRACT

PURPOSE: The aim was to describe the clinical manifestations, complications and long-term outcome of a cohort of Iranian patients with primary immune deficiency (PID). METHOD: We retrospectively studied the demographic, clinical and immunological characteristics of the PID patients in a single tertiary centre, from January 1989 to July 2014. The patients were classified according to the International Union of Immunological Societies Expert Committee on PID. RESULTS: 98 patients were diagnosed with and followed-up for 15 disorders. The mean age at onset and diagnosis and the diagnostic delay were 8±10, 14.2±13.1 and 6.1±7 years, respectively. Parental consanguinity rate was 57%. Predominantly Antibody Deficiency was the most common diagnosis (n=63), followed by congenital defects of phagocytes (n=16), combined immunodeficiencies (n=12), well defined syndromes (n=4) and defects in innate immunity (n=3). Recurrent sinopulmonary infection was the most common presentation. Active infections were treated appropriately, in addition to prophylactic therapy with IVIG and antimicrobials. Not all the patients were compliant with prophylactic regimens due to cost and unavailability. One SCID patient underwent successful bone marrow transplantation. The total mortality rate was 19% during the follow-up period (7.8±7.6 years). The mean age of living patients at the time of study was 23±11.7 years. CONCLUSIONS: Physicians awareness of PID has been rising dramatically in Iran, ensuring an increasing number of patients being diagnosed and treated. More effective treatment services, including health insurance coverage and drug availability are needed to improve the outcome of PID patients.


Subject(s)
Immunoglobulins, Intravenous/therapeutic use , Immunologic Deficiency Syndromes , Immunologic Factors/therapeutic use , Respiratory Tract Infections/epidemiology , Adolescent , Adult , Age of Onset , Child , Child, Preschool , Consanguinity , Delayed Diagnosis , Female , Follow-Up Studies , Humans , Immunoglobulins, Intravenous/administration & dosage , Immunologic Deficiency Syndromes/diagnosis , Immunologic Deficiency Syndromes/epidemiology , Immunologic Deficiency Syndromes/immunology , Immunologic Factors/administration & dosage , Iran/epidemiology , Male , Respiratory Tract Infections/drug therapy , Respiratory Tract Infections/etiology , Respiratory Tract Infections/prevention & control , Retrospective Studies , Tertiary Care Centers/statistics & numerical data , Treatment Outcome , Young Adult
15.
Cell Mol Biol (Noisy-le-grand) ; 61(8): 37-43, 2015 Dec 14.
Article in English | MEDLINE | ID: mdl-26667771

ABSTRACT

Berberine is an isoquinoline alkaloid found in several plant species like famous chinese herb, Rhizoma coptidis which has been used locally as a strong gastrointestinal remedy for thousands of years. The inhibitory effects of berberine on tumor progression properties have been reported before. In this study, we investigated the effect of berberine on an esophageal cancer cell line, KYSE-30 with emphasis on its effects on the expression of certain chemokine receptors. The cytotoxic effect of berberine on KYSE-30 cells was analyzed by MTT assay. In vitro cell migration assay was also applied to the treated cells and the expression levels of the selected chemokine receptors (CXCR4 and CCR7) was measured at mRNA level. A retarded growth, associated with increasing concentrations of berberine, was obvious. On the other hand, the migration rate of the cells was decreased when they were treated with different concentrations of berberine and the expression levels of the two chemokine receptors, involved in the migration and metastasis of esophageal cancer cells, were decreased following the same treatments. With these results, we tend to conclude that berberine might be a proper candidate for further investigations, by targeting the chemokine receptors, and possible applications as anti-metastatic agent in cancer studies.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Berberine/pharmacology , Biomarkers, Tumor/antagonists & inhibitors , Drugs, Chinese Herbal/pharmacology , Gene Expression Regulation, Neoplastic , RNA, Messenger/antagonists & inhibitors , Antineoplastic Agents, Phytogenic/isolation & purification , Berberine/isolation & purification , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/isolation & purification , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Epithelial Cells/pathology , Esophagus/drug effects , Esophagus/metabolism , Esophagus/pathology , Humans , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, CCR7/antagonists & inhibitors , Receptors, CCR7/genetics , Receptors, CCR7/metabolism , Receptors, CXCR4/antagonists & inhibitors , Receptors, CXCR4/genetics , Receptors, CXCR4/metabolism , Signal Transduction
16.
J Postgrad Med ; 61(1): 42-3, 2015.
Article in English | MEDLINE | ID: mdl-25511218

ABSTRACT

Pyoderma gangrenosum (PG) is an uncommon inflammatory ulcerative skin disease. It is characterized by painful progressive necrosis of the wound margins. Rarely, postoperative pyoderma gangrenosum (PPG) manifests as a severe disturbance of wound healing following surgical interventions. Only rare cases of this complication have been reported after appendectomy. We report a case of PPG in a 29-year-old female after appendectomy. She was successfully treated with oral prednisolone. Postoperative pyoderma gangrenosum should be kept in mind in the differential diagnosis of any postoperative delayed wound healing, because this disease is simply distinguished from a postoperative wound.


Subject(s)
Appendectomy , Postoperative Complications/diagnosis , Pyoderma Gangrenosum/complications , Surgical Wound Infection/etiology , Adult , Diagnosis, Differential , Female , Humans , Postoperative Complications/drug therapy , Postoperative Period , Prednisolone/therapeutic use , Pyoderma Gangrenosum/diagnosis , Pyoderma Gangrenosum/drug therapy , Surgical Wound Infection/diagnosis , Surgical Wound Infection/drug therapy , Treatment Outcome
17.
Allergol. immunopatol ; 42(5): 465-471, sept.-oct. 2014. graf, tab
Article in English | IBECS | ID: ibc-127282

ABSTRACT

BACKGROUND: This study was performed to investigate the serum level of interleukin (IL)-13, IL-4, and interferon (IFN)-γ in chronic rhinosinusitis with nasal polyps (CRSwNP) and subsequent inflammation pattern and comorbidities including asthma and aspirin intolerance. METHODS: A case-control study was conducted on 60 adult patients with CRSwNP with mean age of 37.7 ± 12.7 (ranging from 18 to 70) years, and on 20 healthy controls. Serum levels of IL-13, IL-4, and IFN-γ were assessed, using enzyme-linked immunosorbent assay to be compared between case and control groups. Serum level of total immunoglobulin (Ig) E was also assessed in the patients with CRSwNP. RESULTS: Serum level of IL-13 in the patients with CRSwNP was significantly higher than the controls (0.98 ± 1.56 vs. 0.34 ± 0.16 pg/ml, respectively, p = 0.002). IL-4 and IFN-γ did not differ significantly between the two groups. Total IgE level was significantly increased in the patients with CRSwNP, compared to the normal values (301.43 ± 516.54 IU/ml, p = 0.033). Among the patients with CRSwNP, 12/60 (20%) had aspirin intolerance and 44/60 (73.3%) had asthma. IgE was also higher in asthmatics than non-asthmatics patients (364.9 ± 586.6 vs. 126.7 ± 135.7, respectively, p = 0.015). Patients with aspirin intolerance had higher levels of IFN-γ (4.7 ± 1.4 vs. 4.1 ± 0.6, respectively, p = 0.022). CONCLUSIONS: IL-13 with high level of total IgE was observed in the patients with CRSwNP, which predisposes them to have concomitant asthma. IFN-γ seems to be down-regulated in the patients with CRSwNP, but could be over-expressed in the presence of aspirin intolerance


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Subject(s)
Humans , Male , Female , Young Adult , Adult , Middle Aged , Aged , Sinusitis/diagnosis , Sinusitis/immunology , Nasal Polyps/immunology , Nasal Polyps/physiopathology , Asthma/immunology , Interleukins , Interleukins/immunology , Immunoglobulin E/immunology , Case-Control Studies , Lung Volume Measurements , Blood Chemical Analysis/trends
19.
Allergol Immunopathol (Madr) ; 42(5): 465-71, 2014.
Article in English | MEDLINE | ID: mdl-23969075

ABSTRACT

BACKGROUND: This study was performed to investigate the serum level of interleukin (IL)-13, IL-4, and interferon (IFN)-γ in chronic rhinosinusitis with nasal polyps (CRSwNP) and subsequent inflammation pattern and comorbidities including asthma and aspirin intolerance. METHODS: A case-control study was conducted on 60 adult patients with CRSwNP with mean age of 37.7±12.7 (ranging from 18 to 70) years, and on 20 healthy controls. Serum levels of IL-13, IL-4, and IFN-γ were assessed, using enzyme-linked immunosorbent assay to be compared between case and control groups. Serum level of total immunoglobulin (Ig) E was also assessed in the patients with CRSwNP. RESULTS: Serum level of IL-13 in the patients with CRSwNP was significantly higher than the controls (0.98±1.56 vs. 0.34±0.16 pg/ml, respectively, p=0.002). IL-4 and IFN-γ did not differ significantly between the two groups. Total IgE level was significantly increased in the patients with CRSwNP, compared to the normal values (301.43±516.54 IU/ml, p=0.033). Among the patients with CRSwNP, 12/60 (20%) had aspirin intolerance and 44/60 (73.3%) had asthma. IgE was also higher in asthmatics than non-asthmatics patients (364.9±586.6 vs. 126.7±135.7, respectively, p=0.015). Patients with aspirin intolerance had higher levels of IFN-γ (4.7±1.4 vs. 4.1±0.6, respectively, p=0.022). CONCLUSIONS: IL-13 with high level of total IgE was observed in the patients with CRSwNP, which predisposes them to have concomitant asthma. IFN-γ seems to be down-regulated in the patients with CRSwNP, but could be over-expressed in the presence of aspirin intolerance.


Subject(s)
Interferon-gamma/immunology , Interleukin-13/immunology , Interleukin-4/immunology , Nasal Polyps/immunology , Rhinitis/immunology , Sinusitis/immunology , Adolescent , Adult , Aged , Aspirin/adverse effects , Asthma/blood , Asthma/immunology , Case-Control Studies , Drug Hypersensitivity/immunology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoglobulin E/blood , Interferon-gamma/blood , Interleukin-13/blood , Interleukin-4/blood , Male , Middle Aged , Nasal Polyps/blood , Rhinitis/blood , Sinusitis/blood , Young Adult
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