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1.
Arq. bras. med. vet. zootec. (Online) ; 73(4): 989-994, Jul.-Aug. 2021. ilus, mapas
Article in English | LILACS, VETINDEX | ID: biblio-1285275

ABSTRACT

Objetivou-se descrever a ocorrência do Bovine alphaherpesvirus 5 (BoHV5) como causa de meningoencefalite não supurativa em bovinos do estado de Pernambuco, Brasil. Para tanto, 32 amostras de sistema nervoso embebidas em parafina foram obtidas de animais acometidos por doenças neurológicas atendidos na Clínica de Bovinos de Garanhuns da Universidade Federal Rural de Pernambuco (CBG-UFRPE), entre 2012 e 2016. As amostras foram analisadas quanto à presença do gene da glicoproteína C do BoHV5 por reação em cadeia da polimerase (PCR). Dois animais (6,25%) tiveram resultado positivo à PCR, e sua análise de sequenciamento indicou 100% de similaridade para o BoHV5. Os resultados histopatológicos desses dois animais revelaram lesões multifocais de meningoencefalite não supurativa associada à polioencefalomalácia, presença de corpúsculos de inclusão basofílicos, infiltração de células de Gitter e presença de manguitos perivasculares. A PCR se mostra uma importante ferramenta para diferenciação das infecções por BoHV5 de outras enfermidades neurológicas de bovinos, especialmente a raiva.(AU)


Subject(s)
Animals , Cattle , Herpesvirus 5, Bovine/isolation & purification , Meningoencephalitis/veterinary , Paraffin , Central Nervous System , Polymerase Chain Reaction/veterinary , Central Nervous System Viral Diseases/epidemiology
2.
Int. j. morphol ; 38(5): 1197-1200, oct. 2020. graf
Article in English | LILACS | ID: biblio-1134424

ABSTRACT

SUMMARY: Histological techniques are the study of animal and human tissues through staining and examining them under a microscope. To demonstrate the axonal degeneration and demyelination in histological studies, the Luxol Fast Blue staining is gold standard techniques. In this study, a new histochemical method based on modified Luxol Fast Blue for the staining of the myelin sheath in sciatic nerve tissues described. The sciatic nerves of rats were removed and then the sciatic nerve was immersed in 10 % formaldehyde for one week and embedded in paraffin block. Next, thin sections (5 µm) were cut, using a microtome and stained with conventional and modified Luxol Fast Blue. Our results showed that a new method of modified Luxol Fast Blue staining can accurately identify the myelin in the sciatic nerve fibers. The current study showed that the Luxol Fast Blue combination with Light Green has a good effect on myelin coloration, and the results of this study are comparable to LFB combination with Sirius red.


RESUMEN: Las técnicas histológicas son el estudio de tejidos animales y humanos mediante tinción y examen bajo un microscopio. Para demostrar la degeneración axonal y la desmielinización en estudios histológicos, la tinción Luxol Fast Blue es una técnica estándar de oro. En este estudio, se describe un nuevo método histoquímico basado en Luxol Fast Blue modificado para la tinción de mielina en los tejidos del nervio ciático. Se seccionaron los nervios ciáticos de ratas y luego el nervio ciático se sumergió en formaldehído al 10 % durante una semana y se fijó en bloque de parafina. Posteriormente, se cortaron secciones delgadas (5 µm) usando un microtomo y se tiñeron con Luxol Fast Blue convencional y modificado. Nuestros resultados mostraron que un nuevo método de tinción Luxol Fast Blue modificado puede identificar con precisión la mielina en las fibras del nervio ciático. El estudio actual mostró que la combinación Luxol Fast Blue con Light Green es un buen efecto sobre la coloración de mielina, y los resultados de este estudio son comparables a la combinación LFB con Sirius red.


Subject(s)
Animals , Rats , Sciatic Nerve/anatomy & histology , Staining and Labeling/methods , Myelin Sheath , Paraffin , Histological Techniques , Formaldehyde , Microscopy/methods
3.
Article in English | WPRIM | ID: wpr-719428

ABSTRACT

PURPOSE: With the emergence of next-generation sequencing (NGS) technology, profiling a wide range of genomic alterations has become a possibility resulting in improved implementation of targeted cancer therapy. In Asian populations, the prevalence and spectrum of clinically actionable genetic alterations has not yet been determined because of a lack of studies examining high-throughput cancer genomic data. MATERIALS AND METHODS: To address this issue, 1,071 tumor samples were collected from five major cancer institutes in Korea and analyzed using targeted NGS at a centralized laboratory. Samples were either fresh frozen or formalin-fixed, paraffin embedded (FFPE) and the quality and yield of extracted genomic DNA was assessed. In order to estimate the effect of sample condition on the quality of sequencing results, tissue preparation method, specimen type (resected or biopsied) and tissue storage time were compared. RESULTS: We detected 7,360 non-synonymous point mutations, 1,164 small insertions and deletions, 3,173 copy number alterations, and 462 structural variants. Fifty-four percent of tumors had one or more clinically relevant genetic mutation. The distribution of actionable variants was variable among different genes. Fresh frozen tissues, surgically resected specimens, and recently obtained specimens generated superior sequencing results over FFPE tissues, biopsied specimens, and tissues with long storage duration. CONCLUSION: In order to overcome, challenges involved in bringing NGS testing into routine clinical use, a centralized laboratory model was designed that could improve the NGS workflows, provide appropriate turnaround times and control costs with goal of enabling precision medicine.


Subject(s)
Academies and Institutes , Asian Continental Ancestry Group , DNA , Humans , Korea , Methods , Paraffin , Point Mutation , Precision Medicine , Prevalence
4.
Article in English | WPRIM | ID: wpr-764548

ABSTRACT

OBJECTIVES: Human epidermal growth factor receptor-2 (HER2) and 3 (HER3) belong to the epidermal growth factor receptor (EGFR) family of transmembrane receptor tyrosine kinases. In this study, we assessed HER2/HER3 expression levels in specimens of epithelial ovarian cancer and determined their correlation with clinical features of ovarian cancer. METHODS: Tissue microarrays (TMAs) were prepared from paraffin blocks of 105 ovarian tumour samples. HER2, HER3, PI3K, Akt, p-Akt, mTOR, p-mTOR, S6, and p-S6 expression levels were investigated using immunohistochemistry (IHC). HER2 and HER3 amplifications were determined using in situ hybridization (ISH). The correlation between HER2/3 expression and disease outcome of the patients including surgical outcome, progression-free survival (PFS) and overall survival (OS) was analysed. RESULTS: HER2 positivity was 3.8% by IHC and 5.7% by ISH, whereas that of HER3 was 12.4% and 8.6%, respectively. HER2 status by either IHC or ISH was not related to PFS (p=0.128, 0.168, respectively) and OS (p=0.245, 0.164, respectively). However, the HER3 status determined using fluorescence ISH was associated with poor PFS (p=0.035 on log rank test), which was a significant risk factor even after adjusting other possible risk factors in multivariate analysis (hazard ratio=2.377 [1.18–7.49], p=0.021). Expressions of Akt, p-mTOR, and S6 were also related with poor progression (p=0.008, 0.049, 0.014, respectively). CONCLUSION: HER3 is possibly an independent marker for poor prognosis in individuals with ovarian cancer, as the HER3 signalling pathway is distinct from that of HER2. The possibility of targeted therapy for patients with HER3 alteration in ovarian cancer should be evaluated.


Subject(s)
Disease-Free Survival , Epidermal Growth Factor , Fluorescence , Humans , Immunohistochemistry , In Situ Hybridization , Multivariate Analysis , Ovarian Neoplasms , Paraffin , Phosphotransferases , Prognosis , ErbB Receptors , Risk Factors , Tyrosine
5.
Safety and Health at Work ; : 219-223, 2019.
Article in English | WPRIM | ID: wpr-761349

ABSTRACT

BACKGROUND: Ice cooling vests can cause tissue damage and have no flexibility. Therefore, these two undesirable properties of ice cooling vest were optimized, and the present study was aimed to compare the impact of the optimized ice cooling vest and a commercial paraffin cooling vest on physiological and perceptual strain under controlled conditions. METHODS: For optimizing, hydrogel was used to increase the flexibility and a layer of the ethylene vinyl acetate foam was placed into the inside layer of packs to prevent tissue damage. Then, 15 men with an optimized ice cooling vest, with a commercial paraffin cooling vest, and without a cooling vest performed tests including exercise on a treadmill (speed of 2.8 km/hr and slope of %0) under hot (40℃) and dry (40 %) condition for 60 min. The physiological strain index and skin temperature were measured every 5 and 15 minutes, respectively. The heat strain score index and perceptual strain index were also assessed every 15 minutes. RESULTS: The mean values of the physiological and perceptual indices differed significantly between exercise with and without cooling vests (P 0.05). CONCLUSIONS: The optimized ice cooling vest was as effective as the commercial paraffin cooling vest to control the thermal strain. However, ice has a greater latent heat and less production cost.


Subject(s)
Exercise , Hot Temperature , Humans , Hydrogels , Ice , Male , Paraffin , Pliability , Skin Temperature
6.
Article in English | WPRIM | ID: wpr-786126

ABSTRACT

BACKGROUND: Microsatellite instability (MSI) analysis is becoming increasingly important in many types of tumor including colorectal cancer (CRC). The commonly used MSI tests are either time-consuming or labor-intensive. A fully automated MSI test, the Idylla MSI assay, has recently been introduced. However, its diagnostic performance has not been extensively validated in clinical CRC samples.METHODS: We evaluated 133 samples whose MSI status had been rigorously validated by standard polymerase chain reaction (PCR), clinical next-generation sequencing (NGS) cancer panel test, or both. We evaluated the diagnostic performance of the Idylla MSI assay in terms of sensitivity, specificity, and positive and negative predictive values, as well as various sample requirements, such as minimum tumor purity and the quality of paraffin blocks.RESULTS: Compared with the gold standard results confirmed through both PCR MSI test and NGS, the Idylla MSI assay showed 99.05% accuracy (104/105), 100% sensitivity (11/11), 98.94% specificity (93/94), 91.67% positive predictive value (11/12), and 100% negative predictive value (93/93). In addition, the Idylla MSI assay did not require macro-dissection in most samples and reliably detected MSI-high in samples with approximately 10% tumor purity. The total turnaround time was about 150 minutes and the hands-on time was less than 2 minutes.CONCLUSIONS: The Idylla MSI assay shows good diagnostic performance that is sufficient for its implementation in the clinic to determine the MSI status of at least the CRC samples. In addition, the fully automated procedure requires only a few slices of formalin-fixed paraffin-embedded tissue and might greatly save time and labor.


Subject(s)
Colorectal Neoplasms , Microsatellite Instability , Microsatellite Repeats , Paraffin , Polymerase Chain Reaction , Sensitivity and Specificity
7.
Anatomy & Cell Biology ; : 93-96, 2019.
Article in English | WPRIM | ID: wpr-738807

ABSTRACT

Thymus is an encapsulated organ having its bilateral origin from the third pharyngeal pouch. It appears to be a single organ but actually it is bilobed. It attains its maximum development at puberty and then it begins to involute. The parenchyma is replaced by adipocytes and lymphocyte production declines. Here we present a large thymus with a small area of persistent active tissue in it which was obtained during routine undergraduate dissection class. Tissues taken from different quadrants of the large thymic mass were processed, embedded in paraffin and sections were taken for hematoxylin and eosin staining which showed presence of thymic tissue in only one quadrant. Further sections from that quadrant was treated with cytokeratin to confirm its epithelial origin. Therefore knowledge of a large persistent thymus will be helpful to the radiologists and surgeons for making differential diagnosis and in avoiding unnecessary surgical intervention.


Subject(s)
Adipocytes , Adolescent , Aged , Cadaver , Diagnosis, Differential , Eosine Yellowish-(YS) , Hematoxylin , Humans , Immunohistochemistry , Keratins , Lymphocytes , Paraffin , Puberty , Surgeons , Thymus Gland
8.
Article in English | WPRIM | ID: wpr-714688

ABSTRACT

OBJECTIVE: To investigate the expression of androgen receptor (AR) and its correlation with disease status and survival outcome in uterine leiomyosarcoma with other hormone receptors. METHODS: The medical records and paraffin blocks of 42 patients were reviewed. The immunohistochemical expression of AR, estrogen receptor (ER), progesterone receptor (PR), gonadotropin releasing hormone (GnRH), and cytochrome P450, family 19, subfamily A, polypeptide 1 (CYP19A1) were assessed using tissue microarray. RESULTS: In total, AR expression was observed in 11 patients (26.2%). International Federation of Gynecology and Obstetrics (FIGO) stage and AR were independent factors for disease-free survival (DFS) in multivariate regression analysis (odds ratio [OR]=5.8; 95% confidence interval [CI]=1.2–28.4 and OR=0.2; 95% CI=0.05–0.90; p=0.029 and 0.032, respectively). There were no deaths in the AR expression group, whereas the 5-year overall survival (OS) was 54.8% in the no expression group (p=0.014). Co-expression of ER and/or PR with AR was associated with significantly better 5-year DFS and OS than those with negative AR (72.7% vs. 28.6% and 100% vs. 64.3%; p=0.020 and 0.036, respectively). AR may be an independent prognostic marker regardless of ER/PR. CONCLUSION: AR can be a potential prognostic biomarker in uterine leiomyosarcoma.


Subject(s)
Cytochrome P-450 Enzyme System , Disease-Free Survival , Estrogens , Gonadotropin-Releasing Hormone , Gynecology , Humans , Immunohistochemistry , Leiomyosarcoma , Medical Records , Obstetrics , Paraffin , Receptors, Androgen , Receptors, Progesterone
9.
Article in English | WPRIM | ID: wpr-716902

ABSTRACT

OBJECTIVE: To determine whether fragment removal on in vitro fertilization (IVF) day 2 improved the subsequent development and pregnancy outcomes of fragmented embryos compared to similar-grade embryos without fragment removal. METHODS: This study was a retrospective analysis involving 191 IVF cycles in which all embryos had over 10% fragmentation (grade 3 or 4) on day 2 of the IVF-embryo transfer cycle from March 2015 to December 2017. IVF cycles were divided into the fragment removal group (n=87) and the no fragment removal group (n=104) as a control cohort. Before fragment removal, embryos with fragmentation on day 2 were incubated in Ca2+- and Mg2+-free biopsy medium under paraffin oil for 30 minutes. Microsurgical fragment removal was performed with later-assisted hatching and a handmade suction micropipette that had an outer diameter of 30 µm. RESULTS: There were no significant differences in the characteristics of the patients between the control and the fragment removal groups. After fragment removal and subsequent in vitro culture for 24 hours, the number of blastomeres (7.1±1.7 vs. 6.9±1.6) was comparable between the transferred embryos in the two groups, but the morphological grade of the embryos in the fragment removal group (1.9±0.7) was significantly higher than that of the control group (3.1±0.5, p < 0.01). The clinical pregnancy (43.7%) and implantation rates (25.8%) in the fragment removal group were significantly higher than those in the control group (28.8% and 14.0%, respectively; p < 0.05). CONCLUSION: Early fragment removal on day 2 significantly improved the subsequent development and pregnancy outcomes of fragmented embryos.


Subject(s)
Biopsy , Blastomeres , Cohort Studies , Embryonic Structures , Female , Fertilization in Vitro , Humans , In Vitro Techniques , Paraffin , Pregnancy , Pregnancy Outcome , Retrospective Studies , Suction
10.
Article in English | WPRIM | ID: wpr-716864

ABSTRACT

“Paraffinoma” is a well-recognized complication of paraffin oil injection into various body parts for an aesthetic purpose. After a variable latency phase, paraffinoma can present as a wide range of clinical symptoms. This paper is a case report of surgical excision of the paraffinoma and subsequent reconstruction of the associated skin defect on bilateral preauricular cheeks, manifesting 50 years after a primary injection.


Subject(s)
Cheek , Foreign Bodies , Granuloma , Human Body , Paraffin , Skin
11.
Article in Chinese | WPRIM | ID: wpr-775307

ABSTRACT

OBJECTIVE@#To investigate paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples.@*METHODS@#p16 immunohistochemical staining was performed in 150 cases of chronic cervicitis, 126 cases of low-grade squamous intraepithelial lesions(LSIL), 96 cases of high-grade squamous intraepithelial lesions (HSIL) and 78 cases of cervical cancer from January 2014 to March 2018 in Zhongshan Boai Hospital. The results of p16 protein expression in paraffin sections with thickness of 2, 3, 4, 5 and 6 μm were compared using Logistic regression analysis.@*RESULTS@#With the increase of slice thickness, the staining of p16 protein in nucleus gradually increased. The thickness of cervical slices in chronic cervicitis and cervical cancer samples had no significant effect on the positive rate of p16 protein(=7.817 and 1.332, both >0.05), while the thickness of slices in LSIL and HSIL samples had significant effect on the positive rate of p16 protein (=17.688 and 10.182, <0.05 or <0.01). The stable and reliable results were obtained when the slices were between 3 and 5 μm thick.@*CONCLUSIONS@#Paraffin sections with thickness of 3.0-5.0 μm are recommended for immnohistochemical staining of p16 protein in cervical tissue samples.


Subject(s)
Biomarkers, Tumor , Genetics , Metabolism , Cervical Intraepithelial Neoplasia , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , Metabolism , Female , Histocytological Preparation Techniques , Reference Standards , Humans , Immunohistochemistry , Paraffin , Squamous Intraepithelial Lesions of the Cervix , Uterine Cervical Neoplasms
12.
Article in English | WPRIM | ID: wpr-787097

ABSTRACT

In the personalized medicine era, utilizing paraffin blocks in pathology archives for investigating human diseases has come into the limelight. This archived material with clinical data will reduce the research time and could prevent new patient recruitment to obtain tissue for research. However, the clause indicating the necessity of consent from human material providers in the Korean Bioethics and Safety Act has made the Institutional Review Board (IRB) deny permission to use paraffin blocks for research without consent, and alternatively to get the same before starting an experiment. Written consent may be waived off in studies using paraffin blocks with anonymous status or conditions not linked to personal information by applying the paragraph 3, article 16 of the current Bioethics and Safety Act. Also, the IRB should recommend researchers to preserve the blocks as medical records of patients in long-term archives.


Subject(s)
Anonyms and Pseudonyms , Bioethics , Ethics Committees, Research , Humans , Medical Records , Paraffin , Pathology , Patient Selection , Precision Medicine
13.
Article in English | WPRIM | ID: wpr-742113

ABSTRACT

Angiomyolipoma (AML) is a rare benign mesenchymal tumor in the liver, which is composed of blood vessels, smooth muscle, and adipose cells. The proportion of each component varies, making a diagnosis difficult. This paper reports a case of AML in the liver without adipose tissue, mimicking a hepatocellular carcinoma (HCC), which was diagnosed by a surgical tissue biopsy. A 65-year-old woman was admitted for an evaluation of a hepatic mass that had been detected by ultrasonography. The serologic markers of viral hepatitis B and C were negative. The liver function tests and alpha fetoprotein level were within the normal limits. Magnetic resonance imaging revealed a 1.9 cm sized mass in segment 6 of the liver with early arterial enhancement and washout on the delayed phase accompanied by a rim-like enhancement, which is similar to the imaging findings of HCC. A frozen section examination during surgery indicated a hepatocellular neoplasm and suggested the possibility of HCC. On the other hand, the final pathologic diagnosis was epithelioid myoid type of AML with no adipose tissue component. The tumor cells were positive for human melanocyte B-45 and negative for cytokeratin and hepatocyte paraffin 1. This paper reports a very rare case of AML without adipose tissue in the liver mimicking HCC that was diagnosed by a surgical tissue biopsy.


Subject(s)
Adipose Tissue , Aged , alpha-Fetoproteins , Angiomyolipoma , Biopsy , Blood Vessels , Carcinoma, Hepatocellular , Diagnosis , Female , Frozen Sections , Hand , Hepatitis B , Hepatocytes , Humans , Keratins , Liver Function Tests , Liver , Magnetic Resonance Imaging , Melanocytes , Muscle, Smooth , Paraffin , Ultrasonography
14.
Article in English | WPRIM | ID: wpr-741173

ABSTRACT

BACKGROUND: The tumor microenvironment including immune surveillance affects malignant melanoma (MM) behavior. Nuclear factor κB (NF-κB) stimulates the transcription of various genes in the nucleus and plays a role in the inflammatory process and in tumorigenesis. CD8⁺ T cells have cytotoxic properties important in the elimination of tumors. However, inhibitory receptors on the cell surface will bind to programmed death-ligand 1 (PD-L1), causing CD8⁺ T cells to lose their ability to initiate an immune response. This study analyzed the association of NF-κB and PD-L1 expression levels and CD8⁺ T-cell counts with depth of invasion of acral MM, which may be a predictor of aggressiveness related to an increased risk of metastasis. METHODS: A retrospective cross-sectional study was conducted in the Department of Anatomical Pathology, Faculty of Medicine, Universitas Padjadjaran/Hasan Sadikin Hospital using 96 cases of acral melanoma. Immunohistochemical staining was performed on paraffin blocks using anti–NF-κB, –PD-L1, and -CD8 antibodies and invasion depth was measured using dotSlide-imaging software. RESULTS: The study showed significant associations between the individual expression of NF-κB and PD-L1 and CD8⁺ T-cell number, with MM invasion depth. NF-κB was found to be a confounding variable of CD8⁺ T-cell number (p < .05), but not for PD-L1 expression (p = .154). Through multivariate analysis it was found that NF-κB had the greatest association with the depth of invasion (p < .001), whereas PD-L1 was unrelated to the depth of invasion because it depends on the number of CD8⁺ T cells (p = .870). CONCLUSIONS: NF-κB plays a major role in acral MM invasion, by decreasing the number of CD8⁺ T cells in acral MM.


Subject(s)
Antibodies , Carcinogenesis , Cross-Sectional Studies , Melanoma , Multivariate Analysis , Neoplasm Metastasis , Paraffin , Pathology , Retrospective Studies , T-Lymphocytes , Tumor Microenvironment
15.
Article in English | WPRIM | ID: wpr-740152

ABSTRACT

Foreign body injections into breasts may produce foreign body reactions, fibrosis, and local swelling of involved lymph nodes, which can be misdiagnosed as metastasis or malignancy. Here, the authors report MR imaging, PET-CT imaging, and pathologic findings of contralateral internal mammary lymphadenopathy suspicious of breast cancer metastasis in a 58-year-old woman with history of left breast cancer, and previous interstitial mammoplasty by paraffin injection in both breasts.


Subject(s)
Breast Neoplasms , Breast , Female , Fibrosis , Foreign Bodies , Humans , Lymph Nodes , Lymphatic Diseases , Magnetic Resonance Imaging , Mammaplasty , Middle Aged , Neoplasm Metastasis , Paraffin
16.
Arq. bras. med. vet. zootec. (Online) ; 69(6): 1443-1450, nov.-dez. 2017. tab
Article in English | LILACS, VETINDEX | ID: biblio-909839

ABSTRACT

The purpose of the present work was to evaluate the accuracy of quantitative polymerase chain reaction (qPCR) performed on samples of fresh frozen tissue (FT) and formalin-fixed, paraffin-embedded (FFPE) healthy skin. This is a validation study conducted with samples from 46 dogs from an endemic area in Brazil. After sample collection, DNA extractions were conducted using commercial kits and qPCR was oriented to kinetoplast DNA (kDNA) targets of the Leishmania infantum species. The results obtained for the FFPE samples showed 63.6% sensitivity and 77.1% specificity, whereas those obtained for the FT samples showed 100% and 48.6%, respectively. Poor agreement was observed for the results of the qPCR technique with FT and FFPE samples. Our results suggest freezing as the most suitable conservation method for the formation of sample databases considering DNA recovery.(AU)


O objetivo deste trabalho foi avaliar a acurácia da PCR quantitativa (qPCR) realizada em amostras de pele íntegra congelada (FT) e parafinada (FFPE). Trata-se de um estudo de validação, com amostras provenientes de 46 cães de uma área endêmica no Brasil. Após as coletas de amostras, as extrações de DNA foram realizadas utilizando-se kits comerciais, e a qPCR foi orientada para alvos do kDNA da espécie Leishmania infantum. Os resultados obtidos para as amostras FFPE foram 63,6% de sensibilidade e 77,1% de especificidade; para as amostras FT, 100% e 48,6%, respectivamente. A concordância dos resultados da técnica de qPCR com amostras FT e FFPE foi pobre. Os resultados sugerem que o congelamento é o método mais adequado de conservação para banco de amostras para recuperação de DNA.(AU)


Subject(s)
Animals , Dogs , Data Accuracy , Leishmania infantum/isolation & purification , Leishmaniasis, Visceral/diagnosis , Polymerase Chain Reaction/veterinary , Skin/microbiology , Paraffin
17.
Appl. cancer res ; 37: 1-8, 2017. tab, ilus
Article in English | LILACS, Inca | ID: biblio-915391

ABSTRACT

Background: Gastric carcinoma (GC) is the third leading cause of death among malignant tumors worldwide, causing approximately 900,000 deaths/year. Changes in oncogenes that encode tyrosine kinase receptors play an important role in the pathogenesis of GC. MET gene is a proto-oncogene that encodes a tyrosine kinase receptor c-MET and it is required for embryonic development and tissue repair. The hepatocyte growth factor (HGF) is the only known ligand for c-Met receptor. The MET oncogene activation suppresses apoptosis and promotes the survival, proliferation, migration, differentiation and angiogenesis of cells. Among the angiogenic factors, VEGF is the main regulator. Its biological function includes the promotion of endothelial cells mitosis to stimulate cells proliferation. These biomarkers expression in GC is relatively recent and population-based studies are required to define the expression pattern. The aim of this study was to determine qPCR technical standardization to evaluate quantitatively, in paraffin tissue samples, the presence of gene 23 expression of the MET, HGF and VEGF in diffuse and intestinal GC types. Methods: Twenty GC patients were studied, 10 patients were intestinal-type GC (average age 72.1 years) and 10 diffuse-type (average age 50.1 years). In all patients, tissue samples were analyzed from the tumor and distant areas of the tumor tissue. The relative expressions of the tumor markers c-Met, HGF and VEGF were performed by qPCR technique by comparing tumor and non-tumoral samples and they were normalized with the GAPDH constitutive gene. Statistical analysis was performed through T-test. Results: For c-Met, 18/20 (90%) patients expressed the marker and 9/20 (45%) overexpressed this gene, in which three were intestinal-type GC and six were diffuse-type GC. For HGF, only 7/20 (35%) patients expressed this gene and it was overexpressed in 4/20 (20%), in which two were intestinal-type GC and two were diffuse-type GC. For VEGF, 20/20 (100%) patients expressed this marker and in 12/20 (60%) were observed overexpression, in which eight patients had diffuse-type GC and four had intestinal-type GC. Conclusions: qPCR technique was standardized and suitable for expression analysis of the three biomarkers using paraffin embedded tissue samples. Further studies should be carried out to characterize the expression pattern of these biomarkers in GC in the Brazilian population (AU)


Subject(s)
Humans , Male , Female , Paraffin , Stomach , Stomach Neoplasms/genetics , Proto-Oncogenes , Biomarkers, Tumor , Population Control , Proto-Oncogene Proteins c-met , Vascular Endothelial Growth Factor A , Real-Time Polymerase Chain Reaction
18.
Article in English | WPRIM | ID: wpr-29540

ABSTRACT

BACKGROUND AND OBJECTIVES: Neuroinflammation is involved in the pathogenesis of neurodegenerative disorders. Conditioned medium (CM) derived from bone marrow mesenchymal stem cells (MSCs) revealed substantial benefits due to its rich content of trophic factors. Salidroside (Sal), extracted from Rhodiola rosea, is known for its anti-inflammatory and neuroprotective effects. This study was designed to investigate the effect of Sal pretreated CM (CM-Sal) derived from bone marrow MSCs in lipopolysaccharide (LPS) induced neuroinflammation. MATERIAL AND METHODS: Fifty adult male mice were equally divided into 5 groups: Group I (Normal Control), Group II (LPS): single 0.8 mg/kg LPS intraperitoneally; Group III (LPS-DMEM), Group IV (LPS-CM) and Group V (LPS-CM-Sal): LPS was injected as group II followed, 24 hours later, by intranasal injection of 50 μl of filtered serum-free Dulbecco's Modified Eagle's medium (DMEM), CM or CM-Sal, respectively, twice daily for 4 days. Animals were sacrificed at day 6 and paraffin cerebral sections were subjected to Hematoxylin and Eosin staining and immunohistochemistry with caspase 3 (apoptosis), glial fibrillary acidic protein GFAP (astrocytes) and CD68 (active microglia) followed by quantitative morphometric study. RESULTS: Examination of LPS and LPS-DMEM groups revealed neuronal apoptosis with reactive astrogliosis and increased active microglia. LPS-CM and LPS-CM-Sal groups showed less apoptosis, less astrocytes and less active microglia. The regression in neuroinflammation was more evident in LPS-CM-Sal group and the difference was statistically significant compared to other groups. CONCLUSION: CM-Sal derived from MSCs culture elicited significant histopathological improvement in LPS induced neuroinflammation which could be used as new therapeutic modality.


Subject(s)
Adult , Animals , Apoptosis , Astrocytes , Bone Marrow , Caspase 3 , Culture Media, Conditioned , Eosine Yellowish-(YS) , Glial Fibrillary Acidic Protein , Hematoxylin , Humans , Immunohistochemistry , Male , Mesenchymal Stem Cells , Mice , Microglia , Neurodegenerative Diseases , Neurons , Neuroprotective Agents , Paraffin , Rhodiola
19.
Article in English | WPRIM | ID: wpr-17909

ABSTRACT

PURPOSE: This study aimed to analyze G3BP1 and VEZT expression profiles in patients with gastric cancer, and examine the possible relationship between the expressions of each gene and clinicopathological factors. MATERIALS AND METHODS: Expression of these genes in formalin-fixed paraffin embedded (FFPE) tissues, collected from 40 patients with gastric cancer and 40 healthy controls, was analyzed. Differences in gene expression among patient and normal samples were identified using the GraphPad Prism 5 software. For the analysis of real-time polymerase chain reaction products, GelQuantNET software was used. RESULTS: Our findings demonstrated that both VEZT and G3BP1 mRNA expression levels were downregulated in gastric cancer samples compared with those in the normal controls. No significant relationship was found between the expression of these genes and gender (P-value, 0.4835 vs. 0.6350), but there were significant changes associated with age (P-value, 0.0004 vs. 0.0001) and stage of disease (P-value, 0.0019 vs. 0.0001). In addition, there was a direct relationship between VEZT gene expression and metastasis (P-value, 0.0462), in contrast to G3BP1 that did not demonstrate any significant correlation (P-value, 0.1833). CONCLUSIONS: The results suggest that expression profiling of VEZT and G3BP1 can be used for diagnosis of gastric cancer, and specifically, VEZT gene could be considered as a biomarker for the detection of gastric cancer progression.


Subject(s)
Diagnosis , Gene Expression , Humans , Neoplasm Metastasis , Paraffin , Real-Time Polymerase Chain Reaction , RNA, Messenger , Stomach Neoplasms
20.
Asian Spine Journal ; : 863-869, 2017.
Article in English | WPRIM | ID: wpr-102663

ABSTRACT

STUDY DESIGN: Human herniated discs were obtained from discectomy specimens for the immunohistochemical detection of O-GlcNAc and O-GlcNAcase (OGA)/O-GlcNAc transferase (OGT). PURPOSE: This study aimed to quantify the extent of O-GlcNAcylation and its associated enzymes (OGT/OGA) in human degenerated intervertebral discs. OVERVIEW OF LITERATURE: The O-GlcNAcylation of nuclear, cytoplasmic, and mitochondrial proteins as well as the effects of such post-translational modifications are currently the focus of extensive research. O-GlcNAcylation is believed to contribute to the etiology of chronic illnesses by acting as a nutrient and stress sensor in the cellular environment. Mature intervertebral disc cells are chondrocyte-like cells, and O-GlcNAc has been shown to promote chondrocyte apoptosis in vitro. We believe that O-GlcNAcylation is a key regulator of disc degeneration. METHODS: Fifty-six specimens were fixed for 24 hours in a 10% solution of neutral-buffered formaldehyde, dehydrated, and embedded in paraffin. Tissue slices (4-µm-thick) were used for hematoxylin-eosin staining and immunohistochemistry. RESULTS: We found that O-GlcNAcylation of cytoplasmic proteins was less than that of nuclear proteins in both single cells and cell clusters. Cytoplasmic O-GlcNAcylation occurs subsequent to nuclear O-GlcNAcylation and is directly proportional to disc degeneration. OGT and O-GlcNAc expression levels were identical in all specimens examined. CONCLUSIONS: O-GlcNAc and OGA/OGT expression is shown to correlate for the first time with intervertebral disc cell degeneration. Increasing disc degeneration is associated with increasing O-GlcNAcylation in both nuclear and cytoplasmic proteins in human disc cells.


Subject(s)
Apoptosis , Chondrocytes , Chronic Disease , Cytoplasm , Diskectomy , Formaldehyde , Humans , Immunohistochemistry , In Vitro Techniques , Intervertebral Disc Degeneration , Intervertebral Disc Displacement , Intervertebral Disc , Mitochondrial Proteins , Nuclear Proteins , Paraffin , Protein Processing, Post-Translational , Spine , Transferases
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