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1.
Chinese Journal of Biotechnology ; (12): 192-203, 2023.
Article in Chinese | WPRIM | ID: wpr-970368

ABSTRACT

As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163. Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.


Subject(s)
Animals , Swine , Induced Pluripotent Stem Cells/metabolism , Receptors, Cell Surface/genetics , Antigens, CD/metabolism , Porcine respiratory and reproductive syndrome virus/genetics
2.
Int. j. morphol ; 37(2): 606-611, June 2019. tab, graf
Article in English | LILACS | ID: biblio-1002265

ABSTRACT

The purpose of this study was to examine the expression levels of the dental pulp to elucidate the role of Vascular Endothelial Growth Factor (VEGF) and CD68 on vascular angiogenesis, inflammation and odontoblast differentiation in the pulp tissue of diabetic rats depending on the effect of possible damage induced by diabetes. Wistar rats were used in the study, divided into two groups. Control group was fed with standard rat chow and drinking water ad libitum for 8 weeks. Single dose of streptozotocin (STZ) (55 mg/kg), was disolved in sodium citrate buffer and administered by intraperitoneal injection. Blood glucose concentration of rats exceeding 250 mg/dl were accepted as diabetic. Rats were sacrificed under anesthesia. Tissues were immediately dissected, fixed and embedded in paraffin and cut with a microtome then examined under light microscope. In the cross-sections of pulp tissue of diabetic group; the dilation of blood vessels besides hemorrhage and a significant increase in inflammatory cells were seen. The expression of VEGF in the blood vessel endothelial cells of the pulp was increased. VEGF showed positive reaction for degenerative odontoblast cells in the pulp. In this study, increase in VEGF and CD68 expressions in pulp tissue due to the effect of diabetes was thought to delay pulp treatment by inducing soft tissue damage and hypoxia.


El propósito de este estudio fue examinar los niveles de expresión en la pulpa dental para dilucidar el papel del Factor de Crecimiento Endotelial Vascular (VEGF) y el CD68 en la angiogénesis, la inflamación y la diferenciación de odontoblastos en el tejido pulpar de ratas diabéticas, dependiendo del efecto de daño inducido por la diabetes. Se utilizaron ratas Wistar divididas en dos grupos. El grupocontrol se alimentó con comida estándar para ratas y agua potable ad libitum durante 8 semanas. Se administró mediante inyección intraperitoneal dosis única de estreptozotocina (STZ) (55 mg / kg), se disolvió en tampón de citrato de sodio. La concentración de glucosa en sangre de ratas que excedían los 250 mg / dl se aceptó como diabética. Las ratas fueron sacrificadas bajo anestesia. Los tejidos se disecaron de inmediato, se fijaron en parafina y se cortaron para luego ser examinados con un microscopio óptico. En las secciones transversales del tejido pulpar del grupo diabético se observó la dilatación de los vasos sanguíneos además de hemorragia y un aumento significativo de células inflamatorias. La expresión de VEGF se incrementó en las células endoteliales de los vasos sanguíneos de la pulpa. VEGF mostró una reacción positiva para las células odontoblásticas degenerativas en la pulpa. El aumento en la expresión de VEGF y CD68 en el tejido de la pulpa debido al efecto de la diabetes puede retrasar el tratamiento de la pulpa al inducir hipoxia y daños en los tejidos blandos.


Subject(s)
Animals , Male , Rats , Dental Pulp/metabolism , Dental Pulp/pathology , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Experimental/pathology , Immunohistochemistry , Antigens, CD/metabolism , Blotting, Western , Rats, Wistar , Vascular Endothelial Growth Factor A/metabolism , Inflammation , Neovascularization, Pathologic
3.
Braz. j. med. biol. res ; 51(10): e7579, 2018. graf
Article in English | LILACS | ID: biblio-951716

ABSTRACT

Glucocorticoid insensitivity is an important barrier to the treatment of several inflammatory diseases, including acute lung injury (ALI). Saquinavir (SQV) is an inhibitor of the human immunodeficiency virus protease, and the therapeutic effects of SQV in ALI accompanied with glucocorticoid insensitivity have not been previously investigated. In this study, the effects of SQV on lipopolysaccharide (LPS)-mediated injury in human pulmonary microvascular endothelial cells (HPMECs), human type I alveolar epithelial cells (AT I), and alveolar macrophages were determined. In addition, the effects of SQV on an LPS-induced ALI model with or without methylprednisolone (MPS) were studied. In LPS-stimulated HPMECs, SQV treatment resulted in a decrease of high mobility group box 1 (HMGB1), phospho-NF-κB (p-NF-κB), and toll-like receptor 4 (TLR4), and an increase of VE-cadherin. Compared to MPS alone, MPS plus SQV attenuated the decrease of glucocorticoid receptor alpha (GRα) and IκBα in LPS-stimulated HPMECs. HMGB1, TLR4, and p-NF-κB expression were also lessened in LPS-stimulated alveolar macrophages with SQV treatment. In addition, SQV reduced the injury in human AT I with a decrease of HMGB1 and p-NF-κB, and with an increase of aquaporin 5 (AQP 5). SQV ameliorated the lung injury caused by LPS in rats with reductions in vascular permeability, myeloperoxidase activity (MPO) and histopathological scores, and with lowered HMGB1, TLR4, and p-NF-κB expression, but with enhanced VE-cadherin expression. By comparison, SQV plus MPS increased GRα and IκBα in lung tissues of rats with ALI. This study demonstrated that SQV prevented experimental ALI and improved glucocorticoid insensitivity by modulating the HMGB1/TLR4 pathway.


Subject(s)
Animals , Male , Rats , Methylprednisolone/administration & dosage , Saquinavir/administration & dosage , Acute Lung Injury/drug therapy , Signal Transduction/drug effects , Antigens, CD/drug effects , Antigens, CD/metabolism , Cadherins/drug effects , Cadherins/metabolism , Lipopolysaccharides , Rats, Sprague-Dawley , HMGB1 Protein/drug effects , HMGB1 Protein/metabolism , Disease Models, Animal , Toll-Like Receptor 4/drug effects , Toll-Like Receptor 4/metabolism , Acute Lung Injury/chemically induced
4.
An. bras. dermatol ; 91(5): 634-637, Sept.-Oct. 2016. graf
Article in English | LILACS | ID: biblio-827744

ABSTRACT

Abstract: Rosai-Dorfman disease is a benign histiocytic proliferative disorder of unknown etiology. The disease mainly affects lymph node tissue, although it is rarely confined to the skin. Here, we describe a 53-year-old woman with purely cutaneous Rosai-Dorfman disease. The patient presented with a large pigmented plaque on her left leg, and sparse erythematous papules on her face and arms. A complete clinical response was achieved with thalidomide, followed by recurrence at the initial site one year later. The histological examination displayed the typical features of Rosai-Dorfman disease in the recent lesions but not in the older lesions. In the setting of no lymphadenopathy, the histopathological features of Rosai-Dorfman disease are commonly misinterpreted. Therefore, awareness of the histological aspects present at different stages, not always featuring the hallmark microscopic signs of Rosai-Dorfman disease, is particularly important for a correct diagnosis of this rare disorder.


Subject(s)
Humans , Female , Adolescent , Skin Diseases/pathology , Histiocytosis, Sinus/pathology , Arm , Antigens, Differentiation, Myelomonocytic/metabolism , S100 Proteins/metabolism , Antigens, CD/metabolism , Diagnosis, Differential , Histiocytes/pathology , Leg
5.
Braz. j. med. biol. res ; 48(8): 676-682, 08/2015. graf
Article in English | LILACS | ID: lil-753058

ABSTRACT

The function of the visceral yolk sac (VYS) is critical for embryo organogenesis until final fetal development in rats, and can be affected by conditions such as diabetes. In view of the importance of diabetes during pregnancy for maternal and neonatal health, the objective of this study was to assess fetal weight, VYS cell markers, and viability in female Wistar rats (200-250 g) with induced diabetes (alloxan, 37 mg/kg) on the 8th gestational day (gd 8). At gd 15, rats from control (n=5) and diabetic (n=5) groups were anesthetized and laparotomized to remove the uterine horns for weighing of fetuses and collecting the VYS. Flow cytometry was used for characterizing VYS cells, and for determining mitochondrial activity, cell proliferation, DNA ploidy, cell cycle phases, and caspase-3 activity. Fetal weight was reduced in the diabetic group. Expression of the cell markers CD34, VEGFR1, CD115, CD117, CD14, CCR2, CD90, CD44, STRO-1, OCT3/4, and Nanog was detected in VYS cells in both groups. In the diabetic group, significantly decreased expression of CD34 (P<0.05), CCR2 (P<0.001), and OCT3/4 (P<0.01), and significantly increased expression of CD90 (P<0.05), CD117 (P<0.01), and CD14 (P<0.05) were observed. VYS cells with inactive mitochondria, activated caspase-3, and low proliferation were present in the rats with diabetes. Severe hyperglycemia caused by maternal diabetes had negative effects on pregnancy, VYS cell viability, and the expression of cell markers.


Subject(s)
Animals , Male , Female , Pregnancy , Rats , Diabetes Mellitus, Experimental/physiopathology , Pregnancy in Diabetics/physiopathology , Yolk Sac/physiopathology , Antigens, CD/metabolism , Biomarkers/metabolism , Cell Cycle/physiology , Cell Proliferation , Cell Survival , Fetal Weight , Rats, Wistar
6.
Int. j. morphol ; 33(2): 522-526, jun. 2015. ilus
Article in English | LILACS | ID: lil-755504

ABSTRACT

Placental angiogenesis, is essential for embryonic and fetal development. In this study, 18 gestational diabetes mellitus and 22 control pregnancies were included. Gestational diabetes mellitus (GDM) groups compared to the control group significantly higher values were detected (p<0.01). The following histological results were assessed; villous immaturity, chorangiosis, presence of, sncytial knots,mononuclear cell infiltration ischemia and fibrinoid necrosis. To evaluate and compare the placental histology of normal and GDM pregnancies. placentas of pregnant women with gestational diabetes also in terms of angiogenesis and macrophages and ultratructural revealed by examining the possible relationship between fetal complications were investigated.


La angiogénesis de la placenta es esencial para el desarrollo embrionario y fetal. En este estudio, se incluyeron 18 casos de diabetes mellitus gestacional (DMG) y 22 embarazos de control. En grupos los de DMG en comparación con el control, se detectaron valores significativamente mayores (p<0,01) en los siguientes parámetros histológicos que fueron evaluados: inmadurez vellosa, chorangiosis, presencia de nodos sincicial, infiltración celular isquémica mononuclear y necrosis fibrinoide. La investigación de las placentas de mujeres con DMG, reveló mediante el examen en términos de angiogénesis, macrófagos y ultraestructural, la posible relación entre las complicaciones fetales.


Subject(s)
Humans , Female , Pregnancy , Antigens, CD/metabolism , Diabetes, Gestational/metabolism , Diabetes, Gestational/pathology , Placenta/ultrastructure , Vascular Endothelial Growth Factor A/metabolism , Antigens, Differentiation, Myelomonocytic , Immunohistochemistry , Microscopy, Electron , Placenta/metabolism
7.
Biol. Res ; 48: 1-9, 2015. ilus, graf
Article in English | LILACS | ID: biblio-950823

ABSTRACT

BACKGROUND: Theoretically human embryonic stem cells (hESCs) have the capacity to self-renew and differentiate into all human cell types. Therefore, the greatest promise of hESCs-based therapy is to replace the damaged tissues of patients suffering from traumatic or degenerative diseases by the exact same type of cells derived from hESCs. Allo-graft immune rejection is one of the obstacles for hESCs-based clinical applications. Human leukocyte antigen (HLA) II leads to CD4+ T cells-mediated allograft rejection. Hence, we focus on optimizing hESCs for clinic application through gene modification. RESULTS: Transcription activator-like effector nucleases (TALENs) were used to target MHC class II transactivator (CIITA) in hESCs efficiently. CIITA(-/-)hESCs did not show any difference in the differentiation potential and self-renewal capacity. Dendritic cells (DCs) derived from CIITA(-/-)hESCs expressed CD83 and CD86 but without the constitutive HLA II. Fibroblasts derived from CIITA(-/-)hESCs were powerless in IFN-γ inducible expression of HLA II. CONCLUSION: We generated HLA II defected hESCs via deleting CIITA, a master regulator of constitutive and IFN-γ inducible expression of HLA II genes. CIITA(-/-)hESCs can differentiate into tissue cells with non-HLA II expression. It's promising that CIITA(-/-)hESCs-derived cells could be used in cell therapy (e.g., T cells and DCs) and escape the attack of receptors' CD4+ T cells, which are the main effector cells of cellular immunity in allograft.


Subject(s)
Humans , Animals , Mice , Nuclear Proteins/genetics , Trans-Activators/genetics , Cell Differentiation/genetics , Gene Deletion , Deoxyribonucleases/metabolism , Human Embryonic Stem Cells/metabolism , Teratoma , Dendritic Cells/metabolism , Immunoglobulins/metabolism , Immunohistochemistry , Membrane Glycoproteins/metabolism , Tumor Cells, Cultured , Histocompatibility Antigens Class II/genetics , Antigens, CD/metabolism , Interferon-gamma/metabolism , Mice, SCID , Reverse Transcriptase Polymerase Chain Reaction , Deoxyribonucleases/classification , B7-2 Antigen/metabolism , Embryoid Bodies/metabolism , Real-Time Polymerase Chain Reaction , Karyotype , Fibroblasts/metabolism , Cell Self Renewal , Antigen-Presenting Cells/metabolism
9.
Acta cir. bras ; 29(9): 596-602, 09/2014. tab, graf
Article in English | LILACS | ID: lil-722126

ABSTRACT

PURPOSE: To assess the evolution profile of the immunohistochemical expression of stromal constituents over the time-course of wound healing in a murine model. METHODS: Surgical wounds were performed in the back of 24 Wistar rats. After three, seven, 14 and 21 days, six rats were euthanized and the wounded histologically processed to assess the immunohistochemical expression of CD3, CD20, CD31, α-SMA and type-I collagen. Non-injured skin samples (NSS) were used as control. Data were subjected to statistical analysis using ANOVA and Tukey test. RESULTS: The mean of CD3 and CD20 positive cells in the wounds was significantly higher than in NSS at seven and 14 days (p<0.001). The blood vessels content was significantly lower than in NSS (p<0.05) at three days, but increased at seven and 14 days (p<0.01). The mean of α-SMA positive cells at seven, 14 and 21 days was higher than in NSS (p<0.05). The relative content of type I collagen increased from three to 21 days, but remained lower than in NSS (p<0.05). CONCLUSIONS: Lymphoid cells, myofibroblasts and microvessels contents varied over the time-course of wound healing, with peak at seven days and progressive reduction until 21 days. The type I collagen content increased over time. .


Subject(s)
Animals , Male , Actins/metabolism , Antigens, CD/metabolism , Collagen Type I/metabolism , Disease Models, Animal , Lymphocytes/pathology , Skin/injuries , Wound Healing/physiology , /metabolism , /metabolism , /metabolism , Immunohistochemistry , Myofibroblasts/pathology , Rats, Wistar , Skin/metabolism , Stromal Cells/metabolism , Stromal Cells/pathology , Time Factors
10.
Rev. Esc. Enferm. USP ; 48(spe): 137-144, 08/2014. tab, graf
Article in English | LILACS, BDENF | ID: lil-731282

ABSTRACT

Objective To describe the profile of Hospitalizations by Amulatory Care Sensitive Conditions (HACSC), in the Municipality of Cotia, from 2008 to 2012. Method ecological, exploratory, longitudinal study with a quantitative approach. Data on HACSC, by age group and sex, were obtained from the Department of the Unified Health System. For data analysis descriptive statistics were used. Results During the period, there were 46,676 admissions, excluding deliveries, 7,753 (16.61%) by HACSC. The main causes were cerebrovascular diseases, 16.96%, heart failure, 15.50%, hypertension, 10.80% and infection of the kidney and urinary tract, 10.51%. Regarding gender, HACSC occurred predominantly in males. There was a greater number of HACSC at extreme age ranges, especially in the elderly. Conclusion Chronic diseases predominate among the leading causes of HACSC and there was no significant difference between sex.



 .


Objetivo Describir el perfil de las Hospitalizaciones por Condiciones Sensibles de la Atención Primaria (HCSAP), en el municipio de Cotia, entre 2008 y 2012. Método Estudio ecológico, exploratorio, longitudinal con un enfoque cuantitativo. Los datos sobre HCSAP, por grupo de edad y sexo, se obtuvieron del Departamento del Sistema Único de Salud. Para el análisis de los datos se utilizaron estadísticas descriptivas. Resultados Durante el período, hubo 46.676 admisiones, excluyendo entregas, 7.753 (16,61%) por HCSAP. Las principales causas fueron las enfermedades cerebrovascular, 16,96%, insuficiencia cardíaca, 15,50%, hipertensión arterial 10,80% y infección del riñón y las vías urinarias, el 10,51%. Cuanto al género, HCSAP ocurrió mayormente en los hombres. Un mayor número de HCSAP en grupos de edades extremas, especialmente en los ancianos. Conclusión Las enfermedades crónicas predominan entre las principales causas de HCSAP y no hubo diferencia significativa entre los sexo.
 .


Objetivo Descrever o perfil das Internações por Condições Sensíveis à Atenção Primária (ICSAP), no Município de Cotia, entre 2008 e 2012. Método Estudo ecológico, exploratório, longitudinal, de abordagem quantitativa. Dados sobre as ICSAP, segundo a faixa etária e sexo, foram obtidos no Departamento de Informática do Sistema Único de Saúde. Para a análise dos dados foi utilizada a estatística descritiva. Resultados No período, houve 46.676 internações, excluindo os partos, sendo 7.753 (16,61%) por ICSAP. As principais causas foram: doenças cerebrovasculares, 16,96%; insuficiência cardíaca, 15,50%; hipertensão, 10,80%; e infecção do rim e trato urinário, 10,51%. Quanto ao sexo, as ICSAP ocorreram predominantemente nos homens. Houve maior número de ICSAP nos extremos das faixas etárias, especialmente nos idosos. Conclusão As doenças crônicas predominaram entre as principais causas de ICSAP e não houve diferença importante entre os sexos. .


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Antigens, CD/metabolism , /metabolism , Neoplasms/metabolism , Protein-Tyrosine Kinases/metabolism , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/pathology , Focal Adhesion Kinase 1 , Focal Adhesion Protein-Tyrosine Kinases , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Neoplasms/pathology , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , Uterine Neoplasms/metabolism , Uterine Neoplasms/pathology
11.
Journal of Korean Medical Science ; : 846-851, 2014.
Article in English | WPRIM | ID: wpr-163317

ABSTRACT

The purpose of this study was to elucidate the origin and cellular composition of retrocorneal membranes (RCMs) associated with chemical burns using immunohistochemical staining for primitive cell markers. Six cases of RCMs were collected during penetrating keratoplasty. We examined RCMs with hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) staining and immunohistochemical analysis using monoclonal antibodies against hematopoietic stem cells (CD34, CD133, c-kit), mesenchymal stem cells (beta-1-integrin, TGF-beta, vimentin, hSTRO-1), fibroblasts (FGF-beta, alpha-smooth muscle actin), and corneal endothelial cells (type IV collagen, CD133, VEGF, VEGFR1). Histologic analysis of RCMs revealed an organized assembly of spindle-shaped cells, pigment-laden cells, and thin collagenous matrix structures. RCMs were positive for markers of mesenchymal stem cells including beta-1-integrin, TGF-beta, vimentin, and hSTRO-1. Fibroblast markers were also positive, including FGF-beta and alpha-smooth muscle actin (SMA). In contrast, immunohistochemical staining was negative for hematopoietic stem cell markers including CD34, CD133 and c-kit as well as corneal endothelial cell markers such as type IV collagen, CD133 except VEGF and VEGFR1. Pigment-laden cells did not stain with any antibodies. The results of this study suggest that RCMs consist of a thin collagen matrix and fibroblast-like cells and may be a possible neogenetic structure produced from a lineage of bone marrow-derived mesenchymal stem cells.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Antigens, CD/metabolism , Cornea/cytology , Cytokines/metabolism , Endothelial Cells/cytology , Fibroblasts/cytology , Hematopoietic Stem Cells/cytology , Immunohistochemistry , Intercellular Signaling Peptides and Proteins/metabolism , Mesenchymal Stem Cells/cytology , Stem Cells/cytology
12.
An. bras. dermatol ; 88(6): 1005-1007, Nov-Dec/2013. graf
Article in English | LILACS | ID: lil-699011

ABSTRACT

Granular cell tumor is a rare benign neoplasm of neural origin. We report the case of a female patient, 27 years old presenting a brown-red nodule in the right arm, which pathological examination showed to be formed by polygonal cells with eosinophilic granular cytoplasm and immunohistochemistry positive for S100 protein and CD68. Granular cell tumor is usually solitary and in half the cases located in the head and neck areas, 30% of these in the tongue. It is most frequent between the third and fifth decades of life in women and people of African-American ethnicity. Its origination is controversial, including the possible origins in muscle, fibroblasts, neural crest, neural sheath or histiocytes. The positivity for S-100 and CD68 suggest the neural origin.


O tumor de células granulares é uma neoplasia benigna rara, de origem neural. Relatamos caso de paciente feminina, 27 anos, com nódulo de superfície acastanhada no braço direito, cujo exame anatomopatológico evidenciou densa proliferação de células, com amplo citoplasma contendo grânulos eosinofílicos, e imuno-histoquímica positiva para proteínas S100 e CD68. O tumor de células granulares é geralmente solitário e, em metade dos casos, localiza-se em cabeça e pescoço, dos quais 23% na língua. É mais frequente entre a terceira e a quinta décadas de vida, em mulheres e pessoas de etnia negra. A positividade para S-100 e CD68 favorece origem neural.


Subject(s)
Humans , Female , Adult , Skin Neoplasms/pathology , Granular Cell Tumor/pathology , Immunohistochemistry , Antigens, Differentiation, Myelomonocytic/metabolism , S100 Proteins/metabolism , Biomarkers, Tumor/metabolism , Antigens, CD/metabolism
13.
Indian J Exp Biol ; 2013 Feb; 51(2): 107-115
Article in English | IMSEAR | ID: sea-147573

ABSTRACT

Skin stem cells are very important in cosmetics, pharmacological and regenerative medicine and burn cases. Foreskin samples surgically removed after circumcision from boys below 7 years were collected and primary epidermal cells were prepared by enzymatic and mechanical tituration method. Selecting CD133 (prominin-1) multipotent stem cell marker, enriched stem cells were analyzed by MACS using CD133 antibodies conjugated with magnetic beads. CD133 positive and negative cells with specific skin stem cells markers like - CD34 (Universal stem cells marker), CD29 (integrin beta-1) and CD49f (integrin alpha-6) immunophenotypes were screened and sorted in flowcytometer. Further the expression of four embryonic genes or transcription factors of pluripotent stem cells were analyzed for pluripotent character of sorted cells. It was found that skin stem cell markers associated with CD133 cells, differentially expressed CD34, CD29 and CD49f immunophenotyes on both positive and negative CD133 cells in FACS analysis. The embryonic stem cell markers (induced pluripotent stem cell markers) like Oct4, SOX2, Notch-2 and K19 genes were expressed in CD133 positive epidermal cells. It is therefore evident that foreskin derived epidermal stem cells showed pluripotent or multipotent nature. This finding opens up avenues for new uses of these stem cells for direct cell seeding in wound healing, surgical suturing and drug screening.


Subject(s)
Antigens, CD/metabolism , Biomarkers/metabolism , Cell Lineage/genetics , Cell Separation , Cell Survival/genetics , Child , Epidermis/cytology , Flow Cytometry , Gene Expression Profiling , Gene Expression Regulation , Glycoproteins/metabolism , Humans , Immunophenotyping , Male , Peptides/metabolism , Pluripotent Stem Cells/cytology , Pluripotent Stem Cells/metabolism , Propidium/metabolism , Skin/cytology , Staining and Labeling
15.
Gut and Liver ; : 629-641, 2013.
Article in English | WPRIM | ID: wpr-162817

ABSTRACT

Abnormal glycosylation due to dysregulated glycosyltransferases and glycosidases is a key phenomenon of many malignancies, including colorectal cancer (CRC). In particular, increased ST6 Gal I (beta-galactoside alpha 2, 6 sialyltransferase) and subsequently elevated levels of cell-surface alpha 2, 6-linked sialic acids have been associated with metastasis and therapeutic failure in CRC. As many CRC patients experience metastasis to the liver or lung and fail to respond to curative therapies, intensive research efforts have sought to identify the molecular changes underlying CRC metastasis. ST6 Gal I has been shown to facilitate CRC metastasis, and we believe that additional investigations into the involvement of ST6 Gal I in CRC could facilitate the development of new diagnostic and therapeutic targets. This review summarizes how ST6 Gal I has been implicated in the altered expression of sialylated glycoproteins, which have been linked to CRC metastasis, radioresistance, and chemoresistance.


Subject(s)
Humans , Antigens, CD/metabolism , Colorectal Neoplasms/metabolism , Drug Resistance, Neoplasm , Glycoproteins/metabolism , Liver Neoplasms/secondary , Lung Neoplasms/secondary , Radiation Tolerance , ErbB Receptors/metabolism , Sialic Acids/metabolism , Sialyltransferases/metabolism
16.
Experimental & Molecular Medicine ; : e9-2013.
Article in English | WPRIM | ID: wpr-199829

ABSTRACT

Activated protein C (APC) is a cytoprotective anticoagulant that can promote cutaneous healing. We examined the effect of APC on viability and differentiation of the osteoblastic line, MG63, in the presence and absence of bisphosphonates (BPs). Osteoblasts were cultured and treated for 24 or 48 h with Alendronate (Aln), Zoledronate (Zol) or Pamidronate (Pam) at concentrations ranging from 10-4 to 10-6 M. Cell differentiation was measured using type 1 collagen production, Alizarin red staining and alkaline phosphatase activity, whereas cell viability was assessed using MTT and crystal violet assays. All three BPs induced MG63 cell death in a dose- and time-dependent manner. Pam- and Zol-related cell death was prevented by APC treatment; however, cell death induced by Aln was accelerated by APC. APC induced MG63 cell differentiation that was enhanced by Aln, but inhibited by Pam or Zol. Endothelial protein C receptor (EPCR) was expressed by MG63 cells and mediated the protective effect of APC on Zol-induced viability. In summary, we have demonstrated that (1) APC favorably regulates MG63 viability and differentiation toward bone growth, (2) APC differentially regulates the effects of specific BPs and (3) at least part of the effects of APC is mediated through EPCR. These findings highlight the potential importance of the PC pathway in bone physiology and provide strong evidence that APC may influence bone cells and has potential to be a therapeutic drug for bone regeneration, depending on concurrent BP treatment.


Subject(s)
Humans , Antigens, CD/metabolism , Caspases/metabolism , Cell Differentiation/drug effects , Cell Survival/drug effects , Collagen Type I/metabolism , Diphosphonates/pharmacology , Extracellular Signal-Regulated MAP Kinases/metabolism , Matrix Metalloproteinase 2/metabolism , NF-kappa B/metabolism , Osteoblasts/cytology , Protein C/pharmacology , Receptors, Cell Surface/metabolism , Up-Regulation/drug effects
17.
Braz. j. med. biol. res ; 45(3): 197-204, Mar. 2012. ilus, tab
Article in English | LILACS | ID: lil-618047

ABSTRACT

Our group established a method to culture spheres under serum-free culture condition. However, the biological characteristics and the tumorigenicity of spheres are unknown. Here, we demonstrate that sphere cells expressed high levels of the putative colorectal cancer stem cell markers CD133 and CD44. The CD133-positive rates were 13.27 ± 5.62, 52.71 ± 16.97 and 16.47 ± 2.45 percent in sphere cells, regular Colo205 cells and differentiated sphere cells, respectively, while the CD44-positive rates were 62.92 ± 8.38, 79.06 ± 12.10 and 47.80 ± 2.5 percent, respectively, and the CD133/CD44-double-positive rates were 10.77 ± 4.96, 46.89 ± 19.17 and 12.41 ± 2.27 percent, respectively (P < 0.05). Cancer sphere cells formed crypt-like structures in 3-D culture. Moreover, cells from cancer spheres exhibited more tumorigenicity than regular Colo205 cells in a xenograft assay. The cancer sphere cells displayed much higher oncogenicity than regular Colo205 cells to initiate neoplasms, as assayed by H&E staining, Musashi-1 staining and electron microscopy. Our findings indicated that the sphere cells were enriched with cancer stem cells (CSCs), and exhibited more proliferation capacity, more differentiation potential and especially more tumorigenicity than regular Colo205 cells in vitro and in vivo. Further isolation and characterization of these CSCs may provide new insights for novel therapeutic targets and prognostic markers.


Subject(s)
Animals , Humans , Mice , Antigens, CD/metabolism , /metabolism , Cell Proliferation , Colonic Neoplasms/pathology , Glycoproteins/metabolism , Neoplastic Stem Cells/pathology , Peptides/metabolism , Spheroids, Cellular/pathology , Biomarkers, Tumor , Cell Line, Tumor , Cell Culture Techniques/methods , Colonic Neoplasms/metabolism , Mice, Inbred NOD , Mice, SCID , Neoplastic Stem Cells/metabolism , Spheroids, Cellular/metabolism , Tumor Cells, Cultured , Xenograft Model Antitumor Assays
18.
Mem. Inst. Oswaldo Cruz ; 107(2): 170-177, Mar. 2012. graf
Article in English | LILACS | ID: lil-617061

ABSTRACT

Trichomonas vaginalis is a parasite of the human urogenital tract that causes trichomonosis, the most prevalent non-viral sexually transmitted disease. Ectonucleoside triphosphate diphosphohydrolase (NTPDase) family members, which hydrolyse extracellular ATP and ADP and ecto-5′-nucleotidase, which hydrolyses AMP, have been characterised in T. vaginalis. For trichomonad culture, the growth medium is supplemented with 10 percent serum, which is an important source of nutrients, such as adenosine. Here, we investigated the ATP metabolism of T. vaginalis trophozoites from long-term cultures and clinical isolates under limited bovine serum conditions (1 percent serum). The specific enzymatic activities were expressed as nmol inorganic phosphate (Pi) released/min/mg protein, the gene expression patterns were determined by reverse transcriptase-polymerase chain reaction, the extracellular adenine nucleotide hydrolysis was analysed by high performance liquid chromatography and the cell cycle analysis was assessed by flow cytometry. Serum limitation led to the profound activation of NTPDase and ecto-5'-nucleotidase activities. Furthermore, the levels of NTPDase A and B transcripts increased and extracellular ATP metabolism was activated, which led to enhanced ATP hydrolysis and the formation of ADP and AMP. Moreover, the cell cycle was arrested at the G0/G1 stage, which suggested adenosine uptake. Our data suggest that under conditions of serum limitation, NTPDase and ecto-5'-nucleotidase play a role in providing the adenosine required for T. vaginalis growth and that this process contributes to the establishment of parasitism.


Subject(s)
Animals , Cattle , Female , Humans , /metabolism , Adenosine Triphosphate/metabolism , Antigens, CD/metabolism , Apyrase/metabolism , Trichomonas vaginalis/enzymology , Cell Cycle , Chromatography, High Pressure Liquid , Flow Cytometry , Reverse Transcriptase Polymerase Chain Reaction
19.
The Korean Journal of Gastroenterology ; : 325-329, 2012.
Article in Korean | WPRIM | ID: wpr-11954

ABSTRACT

Collision tumors of the colon are rare. A 54-year-old man was referred to our hospital for the evaluation of hematochezia. Colonoscopy demonstrated the presence of about 3 cm sized mass in the rectosigmoid junction. After surgical resection, the colonic lesion was histologically composed of two discrete lesions: adenocarcinoma in the superficial layer and poorly differentiated neuroendocrine carcinoma in the deeper layer. We report this case of colonic collision tumor (adenocarcinoma and neuroendocrine carcinoma) with a review of the literature.


Subject(s)
Humans , Male , Middle Aged , Adenocarcinoma/diagnosis , Antigens, CD/metabolism , Antigens, Differentiation, T-Lymphocyte/metabolism , Carcinoma, Neuroendocrine/diagnosis , Colonic Neoplasms/diagnosis , Colonoscopy , Positron Emission Tomography Computed Tomography , Synaptophysin/metabolism , Tomography, X-Ray Computed
20.
Journal of Korean Medical Science ; : 893-899, 2011.
Article in English | WPRIM | ID: wpr-31559

ABSTRACT

Thyroid carcinogenesis is accompanied by loss of thyroid-specific functions and refractory to radioiodine and thyroid stimulating hormone (TSH) suppression therapy. Redifferentiating agents have been shown to inhibit tumor growth and improve the response to conventional therapy. Polyphenol phytochemicals (PPs) in fruits and vegetables have been reported to inhibit cancer initiation, promotion, progression and induce redifferentiation in selected types. In this study we examined PPs induce redifferentiation in thyroid cancer cell lines. We investigated the effects of genistein, resveratrol, quercetin, kaempferol, and resorcinol on the F9 embryonal carcinoma cell differentiation model. The thyroid cancer cell lines, TPC-1, FTC-133, NPA, FRO, and ARO, displayed growth inhibition in response to genistein, resveratrol, quercetin. We further demonstrated that genistein decreased the dedifferention marker CD97 in NPA cells and resveratrol decreased CD97 in FTC-133, NPA, FRO cells and quercetin decreased CD97 in all cell lines. We observed increased expression of differentiation marker NIS in FTC-133 cells in response to genistein, and resveratrol but no change in NPA, FRO, ARO cells. Quercetin increased or induced NIS in FTC-133, NPA, FRO cells. These findings suggest that PPs may provide a useful therapeutic intervention in thyroid cancer redifferentiation therapy.


Subject(s)
Humans , Antigens, CD/metabolism , Antineoplastic Agents/pharmacology , Carcinoma, Embryonal/drug therapy , Cell Differentiation/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Flavonoids/pharmacology , Gene Expression Regulation, Neoplastic , Genistein/pharmacology , Kaempferols/pharmacology , Models, Biological , Phenols/pharmacology , Quercetin/pharmacology , Resorcinols/pharmacology , Stilbenes/pharmacology , Symporters/metabolism , Thyroid Neoplasms/drug therapy
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