ABSTRACT
This work aimed to assess the effect of the probiotic strain, Lactobacillus plantarum, on the levels of leptin, IGF-1 and their receptors on the hepatopancreatic tissues of Nile tilapia (Oreochromis niloticus) and then correlate fish growth performance and gut microbiological parameters. Fish juveniles (±23g) were reared in a recirculation system with constant aeration and temperature (25°C). They were distributed into six polyethylene tanks (45L) and fed twice a day at 5% of the tank biomass with the respective diets: control (commercial diet without probiotic) and supplemented with L. plantarum inoculum (1 x 108 CFU mL-1), both in triplicate. After 30 days of feeding, L. plantarum-fed fishes showed greater weekly growth rate, final weight, and feed conversion rate, in addition to higher count of lactic-acid bacteria and lower count of pathogenic bacteria in the intestinal tract, when compared to the control group. The immunostaining intensity for IGF-1 and leptin hormones was lower after L. plantarum supplementation than in the control group, with no change in the level for receptors. This reduction could implicate important changes in fish metabolism and homeostasis.(AU)
O presente trabalho teve como objetivo avaliar o efeito da cepa probiótica Lactobacillus plantarum sobre os níveis de leptina, IGF-1 e seus receptores no tecido hepatopancreático de tilápia-do-nilo (Oreochromis niloticus) e correlacionar com o desempenho zootécnico e os parâmetros microbiológicos intestinais dos peixes. Juvenis de tilápia-do-nilo (±23g) foram distribuídos em seis tanques de polietileno (45L) conectados a um sistema de recirculação, com aeração e temperatura constantes (25°C). Os peixes foram alimentados duas vezes ao dia, a 5% da biomassa do tanque, com as respectivas dietas: controle (dieta comercial sem probiótico) e suplementada com L. plantarum (1 x 108 UFC mL-1), ambas em triplicata. Após 30 dias de cultivo, os peixes alimentados com L. plantarum apresentaram maiores ganho de peso semanal, peso final e conversão alimentar, bem como maior contagem de bactérias ácido-láticas e menor contagem de bactérias patogênicas no trato intestinal das tilápias alimentadas com dieta probiótica, em comparação ao grupo controle. A intensidade da imunomarcação para os hormônios IGF-1 e leptina foi menor com a suplementação de L. plantarum do que no grupo controle, sem alterar os níveis de seus receptores. Essa redução pode implicar mudanças importantes no metabolismo e na homeostase dos peixes.(AU)
Subject(s)
Animals , Cichlids/growth & development , Hepatopancreas/chemistry , Lactobacillus plantarum , Gastrointestinal Microbiome , Animal Feed , Insulin-Like Growth Factor I , Dietary Supplements , LeptinABSTRACT
Cryptorchidism is associated with infertility in adulthood. Early orchiopexy is suggested to reduce the risk. Information is lacking on the potential link between infant germ cell maturation and the risk of future infertility. The objective of the study was to evaluate age-related germ cell development in cryptorchidism. Immunostaining for markers of germ cell development (octamer-binding transcription factor 3/4 [OCT3/4], placental alkaline phosphatase [PLAP], KIT proto-oncogene [C-KIT], podoplanin [D2-40], Lin-28 homolog A [LIN28], and G antigen 7 [GAGE-7]) was performed in testicular biopsies from 40 cryptorchid boys aged 4-35 months. Germ cell numbers and distributions were evaluated in cross sections of seminiferous tubules, with and without immunostaining. OCT3/4, D2-40, and LIN28 were generally expressed in the early stages of germ cell development, as shown by positive expression in germ cells in the central region of seminiferous tubules. In contrast, PLAP and GAGE-7 were expressed in both central and peripheral parts of the tubules in the early stages of development and expressed mainly in a peripheral position with advancing age. Germ cell maturation was delayed in this study population as compared with that observed in our previous study on germ cell markers in a healthy population. The number of GAGE-7-positive germ cells per tubular cross section obtained by immunostaining was significantly higher than that obtained by standard hematoxylin and eosin staining. Double immunostaining revealed heterogeneity in germ cell development in cryptorchid testes. These results shed light on the pathophysiology of germ cell development in boys with cryptorchidism.
ABSTRACT
Introduction: In the present era, FNAC has proved to be anessential primary diagnostic procedure for soft tissue lesions.To correlate its efficacy and to further subtype,histopathological examination is done aided by IHC if required.The present study aimed to evaluate the epidemiologicaldistribution of soft tissue lesions with reference to age, sex andsite and to assess the utility of FNAC in terms of sensitivity,specificity, positive and negative predictive values, and overallhistological correlation percentage of cytology in diagnosingvarious types of soft tissue lesions.Materials and Methods: Prospective study was carried outduring the period of Sept 2017 to May 2019 for FNAexamination of soft tissue lesions. Cytopathological andhistopathological examination was carried out in all cases withimmunostaining done in few cases.Results: Of 463 soft tissue lesions that could be successfullyfollowed up, 347 were benign lesions and rest were malignant.Most common age group affected were 31-40 years with slightmale preponderance (M:F=1.37:1). Most common site beingLower extremities. Lipomas were the most common soft tissuelesions (169 cases) and spindle cell sarcomas were the mostcommon malignant lesions. The cytological andhistopathological diagnosis correlated well in almost all casesexcept discordance was seen in 5 cases. The sensitivity andspecificity of the procedure were 97.4% and 99.4%respectively.Conclusion: FNAC was found to be a highly specific andsensitive tool in diagnosing soft tissue lesions and can be fairlyimplemented as it is well tolerated and cost effective forpatients.
ABSTRACT
@#We present a case of an undifferentiated subtype of non-keratinizing squamous cell carcinoma (NK-SCC) with sarcomatoid features in the nasopharynx in a 69-year-old man who was difficult to diagnose due to spindle-shaped malignant cells. He was admitted because of a right nasal obstruction and right headache, and imaging revealed a heterogeneously enhanced irregularly shaped mass at the nasopharynx. Histopathologically, the tumour was partially organised, and the tumour cells were epithelioid or spindle-shaped. Initially, we erroneously diagnosed the tumour as an angiosarcoma owing to its false-negative immunoreaction for cytokeratins and a mistaken interpretation for CD31. After in situ hybridization for Epstein-Barr virus was positive, a consultation and additional immunostaining (including re-staining for cytokeratin with varying dilutions) were performed, and the diagnosis was revised to NK-SCC with sarcomatoid features. We believe that sarcomatoid features may be observed in nasopharyngeal carcinoma and in this case, immunostaining using various epithelial markers is necessary and careful attention should be paid to the interpretation of immunostaining.
Subject(s)
Nasopharyngeal CarcinomaABSTRACT
Leprosy is a chronic granulomatous disease affecting skin, peripheral nerves and other tissues. On histopathology leprosy mimics other infectious and non-infectious lesions like tuberculosis, sarcoidosis and fungal infections, which are also common in our country. In tuberculoid and indeterminate forms, where Acid Fast Bacilli cannot be demonstrated, the diagnosis becomes more difficult. Mycobacterium leprae is the only bacterium which has the ability to infiltrate peripheral nerves leading to Schwann cell disintegration. On routine Hematoxylin and Eosin stains (H&E), the nerve fibers may not be easily identifiable in some cases , hence S-100 immunostaining is used to highlight the nerve elements and to demonstrate and compare the nerve changes in spectrum of leprosy. With widespread use of multi-drug treatment, there has been changes in the profile of disease. The aim of the present study was to observe different patterns of cutaneous nerve involvement in leprosy and to correlate these with the clinical and histopathological findings in currently referred cases for histopathological opinion. The study was conducted in the Department of Pathology, Himalayan Institute of Medical Sciences, Swami Rama Himalayan University, Dehradun, over a period of 12 months (July 2016 - July 2017) Subjects were recruited from patients presenting in Dermatology OPD. A total 35 consecutive cases with clinical suspicion / diagnosis of leprosy were included in the study. Biopsies were processed and stained by H&E, Fite-Faraco as well as S100 immunostaining. It was observed that on S-100 immunostaining, 43.7% cases showed granulomas infiltrating the dermal nerves whereas these changes could not be demonstrated in 16.6% cases of Borderline leprosy on H&E staining alone. Thus S-100 staining appears to serve as an important tool to diagnose leprosy from other granulomatous diseases of skin even in current scenario of leprosy.
ABSTRACT
Given the limited studies and conflicting findings, the transport character of ginsenosides crossing the blood-brain barrier (BBB) remains unclear. The present study was designed to qualitatively determine the distribution of ginsenosides in brain tissues after oral administration of ginseng total saponins, using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) combined with immunohistochemistry. In brain tissue homogenates, ginsenoside Rg1 was detectable and no other ginsenosides or their metabolites were found. No ginsenosides were detected in cerebrospinal fluid. Immunohistochemistry staining of brain tissue sections by using anti-ginsenoside polyclonal antibodies revealed the localization of ginsenosides in brain tissues. Furthermore, immunofluorescence double staining revealed that ginsenosides widely existed in vascular endotheliocytes and astrocytes, and in few neurons. These results indicated that Rg1 was the main component that entered the brain after oral administration of ginseng total saponins and that ginsenosides could cross the BBB, although the transport capability of ginsenosides through the BBB may be poor.
Subject(s)
Animals , Male , Mice , Rats , Administration, Oral , Antibodies , Blood-Brain Barrier , Metabolism , Brain , Metabolism , Brain Chemistry , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Metabolism , Ginsenosides , Metabolism , Mice, Inbred C57BL , Panax , Chemistry , Rats, Sprague-Dawley , Tandem Mass SpectrometryABSTRACT
Inhibitory GABAergic interneurons are fundamental elements of cortical circuits and play critical roles in shaping network activity. Dysfunction of interneurons can lead to various brain disorders, including epilepsy, schizophrenia, and anxiety. Based on the electrophysiological properties, cell morphology, and molecular identity, interneurons could be classified into various subgroups. In this study, we investigated the density and laminar distribution of different interneuron types and the co-expression of molecular markers in epileptic human cortex. We found that parvalbumin (PV) and somatostatin (SST) neurons were distributed in all cortical layers except layer I, while tyrosine hydroxylase (TH) and neuropeptide Y (NPY) were abundant in the deep layers and white matter. Cholecystokinin (CCK) neurons showed a high density in layers IV and VI. Neurons with these markers constituted ~7.2% (PV), 2.6% (SST), 0.5% (TH), 0.5% (NPY), and 4.4% (CCK) of the gray-matter neuron population. Double- and triple-labeling revealed that NPY neurons were also SST-immunoreactive (97.7%), and TH neurons were more likely to express SST (34.2%) than PV (14.6%). A subpopulation of CCK neurons (28.0%) also expressed PV, but none contained SST. Together, these results revealed the density and distribution patterns of different interneuron populations and the overlap between molecular markers in epileptic human cortex.
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Brain Chemistry , Genetics , Physiology , Cerebral Cortex , Metabolism , Pathology , Cholecystokinin , Metabolism , Epilepsy , Pathology , Gene Expression Regulation , Physiology , Interneurons , Metabolism , Neuropeptide Y , Metabolism , Parvalbumins , Metabolism , Phosphopyruvate Hydratase , Metabolism , Somatostatin , Metabolism , Tyrosine 3-Monooxygenase , MetabolismABSTRACT
Given the limited studies and conflicting findings, the transport character of ginsenosides crossing the blood-brain barrier (BBB) remains unclear. The present study was designed to qualitatively determine the distribution of ginsenosides in brain tissues after oral administration of ginseng total saponins, using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) combined with immunohistochemistry. In brain tissue homogenates, ginsenoside Rg1 was detectable and no other ginsenosides or their metabolites were found. No ginsenosides were detected in cerebrospinal fluid. Immunohistochemistry staining of brain tissue sections by using anti-ginsenoside polyclonal antibodies revealed the localization of ginsenosides in brain tissues. Furthermore, immunofluorescence double staining revealed that ginsenosides widely existed in vascular endotheliocytes and astrocytes, and in few neurons. These results indicated that Rg1 was the main component that entered the brain after oral administration of ginseng total saponins and that ginsenosides could cross the BBB, although the transport capability of ginsenosides through the BBB may be poor.
Subject(s)
Animals , Male , Mice , Rats , Administration, Oral , Antibodies , Blood-Brain Barrier , Metabolism , Brain , Metabolism , Brain Chemistry , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Metabolism , Ginsenosides , Metabolism , Mice, Inbred C57BL , Panax , Chemistry , Rats, Sprague-Dawley , Tandem Mass SpectrometryABSTRACT
BACKGROUND: BRCA1-associated protein 1 (BAP1) mutations are frequently reported in clear cell renal cell carcinoma (ccRCC); however, very few studies have evaluated the role of these mutations in other renal cell carcinoma (RCC) subtypes. Therefore, we analyzed BAP1 protein expression using immunohistochemistry in several RCC subtypes and assessed its relationship with clinicopathological characteristics of patients. METHODS: BAP1 expression was immunohistochemically evaluated in tissue microarray blocks constructed from 371 samples of RCC collected from two medical institutions. BAP1 expression was evaluated based on the extent of nuclear staining in tumor cells, and no expression or expression in < 10% of tumor cells was defined as negative. RESULTS: Loss of BAP1 expression was observed in ccRCC (56/300, 18.7%), chromophobe RCC (6/26, 23.1%), and clear cell papillary RCC (1/4, 25%), while we failed to detect BAP1 expression loss in papillary RCC, acquired cystic disease-associated RCC, or collecting duct carcinoma. In ccRCC, loss of BAP1 expression was significantly associated with high World Health Organization (WHO)/International Society of Urological Pathology (ISUP) grade (p = .002); however, no significant correlation was observed between loss of BAP1 expression and survival in ccRCC. Loss of BAP1 expression showed no association with prognostic factors in chromophobe RCC. CONCLUSIONS: Loss of BAP1 nuclear expression was observed in both ccRCC and chromophobe RCC. In addition, BAP1 expression loss was associated with poor prognostic factors such as high WHO/ISUP grade in ccRCC.
Subject(s)
Humans , Carcinoma, Renal Cell , Immunohistochemistry , Pathology , World Health OrganizationABSTRACT
CD10, a transmembrane endopeptidase, has been shown to be lost as an early event in prostate cancer. We aimed at evaluating the pattern of expression of CD10 in various Gleason’s grades of prostatic adenocarcinoma in comparison with nodular hyperplasia of prostate. This retrospective study included 30 cases of nodular hyperplasia and 30 of prostatic adenocarcinoma of various Gleason’s grades. Immunohistochemical staining for CD10 was performed on all cases and positivity evaluated as percentage of cells as well as location (membranous or cytoplasmic or both). Of prostatic adenocarcinomas, grade 3 was seen in 10 foci, grade 4 in 28 and grade 5 in 22 foci. CD10 positivity in carcinoma was lower than in nodular hyperplasia, with the lowest positivity in grade 5. The pattern of expression of CD10 also changed from membranous in grade 3 to cytoplasmic in grade 5. Loss of CD10 expression appears to be associated with increasing tumour grade in carcinoma prostate and this can potentially be useful in stratification of such patients.
ABSTRACT
Epidermal growth factor receptor (EGFR) is a 170-kDa tyrosine kinase transmembrane glycoprotein expressed in normal tissues in many organs and different types of tumours. In prostate, EGFR is expressed mainly in epithelial cells, phosphorylation of EGFR (pEGFR) which is assessed by immunohistochemical methods could be useful prognostic marker for prostate cancer cases. Tumours may affect the surrounding non-malignant tissue and pEGFR immunoreactivity in the morphologically normal prostate tissue can be used to retrieve prognostic information. In this study the membranous and cytoplasmic expression of EGFR is checked in both the basal and luminal cells. Intensity of the staining and the pattern of the staining were noted in benign, in-situ and malignant lesions and it was found that the staining intensity of the luminal cells increase with a subsequent decreased staining in the basal layer as the lesion progress towards malignancy. Subsequently the staining intensity and patterns were correlated with the Gleason grade for triaging of the cases into different prognostic groups.
ABSTRACT
Introducción: se define a las leucoplasias orales como una placa blancaque no puede desprenderse por raspado y que no puede clasifi carse comoninguna otra lesión. Son lesiones con potencial maligno, relacionadascon la presencia de displasia epitelial. Estos cambios preneoplásicospueden ser evidenciados histológicamente como también a travésde técnicas que pongan en evidencia los diferentes cambios a nivelmolecular. La E-cadherina es una glicoproteína membranosa quedesempeña papeles importantes en el mantenimiento de la adhesióncélula-célula, la preservación de la polaridad del tejido epitelial y laintegridad estructural. Los factores de crecimiento epidérmico son unconjunto de moléculas de naturaleza proteica, biorreguladores, cuyafuncionalidad fundamental radica en el control del ciclo celular. Elobjetivo del presente trabajo es identifi car y comparar parámetros histológicosy moleculares predictores de riesgo de transformación malignaen leucoplasias orales. Material y métodos: El estudio correspondea un diseño observacional descriptivo. Se seleccionaron muestras de26 biopsias de leucoplasias orales, las cuales fueron evaluadas contécnica histológica de rutina y tinción con hematoxilina y eosina, luegosometidas a inmunomarcación con factor de crecimiento epidérmico yE-cadherina, donde se evaluó la intensidad de tinción y cambios en laexpresión de cada marcador, así como la localización en los diferentessubtipos celulares. Resultados: De las 26 leucoplasias observadas,16 mostraron histología con cambios hiperplásicos y 10 con cambiosdisplásicos leves a moderados. La expresión de E-cadherina no mostróalteraciones signifi cativas en leucoplasias sin displasia, sólo hubopérdida de expresión en aquellas leucoplasias con cambios displásicosde alto grado, en concordancia a los hallazgos histológicos...
Introduction: oral leukoplakia is defined as a white plaque thatcannot be removed by scraping and cannot be classifi ed as any otherdisease entity. They are potentially malignant lesions related to thepresence of epithelial dysplasia. These preneoplastic changes can bedetected histologically, as well as through techniques that demonstratediff erent changes at the molecular level. E-cadherin is a membraneglycoprotein that plays a major role in maintaining cell-cell adhesion,preserving structural integrity and the polarity of epithelial tissue.Epidermal growth factors are a group of bio-regulatory proteins,whose primary function is to control the cell cycle. The aim of thisstudy is to identify and compare the parameters for histological andmolecular markers for malignant transformation in oral leukoplakia.Material and methods: The study was observational and descriptive indesign. Samples were selected from 26 oral leukoplakia biopsies, whichwere routinely evaluated for histology and stained with hematoxylinand eosin, then subjected to immunostaining with epidermal growthfactor and E-cadherin, with the intensity of staining and changes inthe expression of each marker being evaluated. Results: Of the 26leukoplakia examined, 16 showed hyperplastic changes and 10 mildto moderate dysplastic changes. The expression of E-cadherin showedno signifi cant changes in non-dysplastic leukoplakia, while a lossof expression was found in only those leukoplakias with high-gradedysplastic changes, which was consistent with the histological fi ndings...
Subject(s)
Humans , Male , Female , Cadherins/physiology , Epidermal Growth Factor/immunology , Precancerous Conditions/etiology , Leukoplakia, Oral/immunology , Argentina , Biopsy/methods , Cell Transformation, Neoplastic , Epidemiology, Descriptive , Immunohistochemistry/methods , Observational Studies as Topic , Data Interpretation, StatisticalABSTRACT
Introduction: Decoy receptor 3 (DcR3) functions as a death decoy inhibiting apoptosis mediated by the tumor necrosis factor receptor family. It is highly expressed in many tumors and its expression can be regulated by the MAPK/ ERK signaling pathway and ERK is a vital member of this pathway. Toll‑like receptor 4 (TLR4) is expressed on immune cells. Increased TLR4 expression has been associated with various types of cancers. Material and Methods: The study was conducted to investigate the expression of DcR3, ERK1/2, and TLR4 in astrocytomas and evaluate if they are validating markers for discriminating glioblastoma from anaplastic astrocytoma in limited surgical specimen. Expression of DcR3, ERK1/2, and TLR4 was determined by immunohistochemical staining of tissue microarray from 48 paraffin‑embedded tissues. A binary logistic regression method was used to generate functions that discriminate between anaplastic astrocytomas and glioblastomas. Results: The expression of TLR4 and DcR3 was significantly higher in glioblastomas than in anaplastic astrocytomas. DcR3 could discriminate anaplastic astrocytomas from glioblastomas with high sensitivity (93.8%), specificity (90%), and accuracy (92.3%). Conclusion: Our results suggest that DcR3 may be a useful marker for discriminating anaplastic astrocytomas from glioblastomas.
ABSTRACT
Background & objectives: Renal tumours constitute about 7 per cent of all neoplasms in children. It is important to differentiate Wilms’ tumour (commonest tumour) from non-Wilms’ tumours. The aim of this study was to evaluate the immunoexpression and diagnostic role of Wilms’ tumour-1 protein (WT1) in paediatric renal tumours. Methods: A total of 53 cases of renal tumours in children (below 18 yr) who underwent total nephrectomy were included in this retrospective study. WT1 immunostaining was done using mouse monoclonal WT1 antibody (clone: 6F-H2). Results: Of the 53 cases, 38 (72%) were of Wilms’ tumour. Non-Wilms’ group (15) included six cases of mesoblastic nephroma (MN), two each of clear cell sarcoma (CCSK), renal cell carcinoma (RCC) and peripheral neuroectodermal tumour (PNET) and one each of angiomyolipoma (AML), rhabdomyosarcoma (RMS) and malignant rhabdoid tumour (MRT). Proportion of WT1 positivity in Wilms’ tumour was 100 per cent in contrast to 26.7 per cent in non-Wilms’ tumours (P<0.001). Epithelial and blastemal components of Wilms’ tumour showed moderate (2+) nuclear and cytoplasmic staining in 80 (24/30) and 75 per cent (24/32) cases, respectively. MN, PNET, CCSK and AML were negative for WT1. RMS, RCC and MRT showed cytoplasmic staining, strongest in RMS. No significant association was seen between WT1 expression and NWTSG (National Wilms’ Tumor Study Group) stage. Interpretation & conclusions: WT1 helps to differentiate Wilms’ tumour from other paediatric renal tumours. It may help in differentiating the two subgroups of Wilms’ tumour which have distinct molecular pathogenesis and biological behaviour, however, further prospective studies are required for validation of this hypothesis.
ABSTRACT
The respiratory epithelium is the first line of contact with the external hazards. Thus it can be damaged and need to be replaced to avoid healing by fibrosis. Tracheal tissue engineering is an alternative promising treatment modality. Mesenchymal stem cell markers are surface proteins, which are responsible for some of these cells unique properties. The objective of this study was to detect the mesenchymal stem cell phenotype among the human nasal respiratory epithelial cells via two immunophenotyping techniques. Respiratory epithelial cells were cultured using co-culture technique, fibroblasts was removed at confluence leaving respiratory epithelial cells, which were passage further to passage 4. Cells were evaluated for mesenchymal stem cell markers that were CD73, CD90, CD105 and the hematopoietic stem cell marker CD45 at passage 1 (P1) and passage 4 (P4) using Flow cytometry and Immunocytochemistry techniques. Respiratory epithelial cells expressed the mesenchymal stem cell markers at P1 and maintain the expression these markers until P4. Using both techniques, to compare the values of mesenchymal stem cell markers expression at P1 to P4 there was no significant difference. This study indicates that respiratory epithelial cells derived from nasal turbinate retain some of mesenchymal stem cells properties even after serial passages. Both methods of Immunophenotyping are comparable.
El epitelio respiratorio es la primera línea de contacto con los peligros externos. Por lo tanto, puede ser dañado y necesita ser reemplazado para evitar uan cicatrización por fibrosis. La ingeniería de tejidos traqueales es una modalidad de tratamiento alternativo prometedora. Los marcadores de células troncales mesenquimales son proteínas de superficie, que son responsables de algunas propiedades únicas de estas células. El objetivo fue detectar el fenotipo de células troncales mesenquimales entre las células epiteliales respiratorias nasales humanas a través de dos técnicas de inmunofenotipaje. Fueron cultivadas las células epiteliales respiratorias utilizando la técnica de co-cultivo; los fibroblastos se eliminaron en la confluencia dejando solo células epiteliales respiratorias, resultantes de los 4 pasajes. Las células fueron evaluadas para encontrar marcadores de células troncales mesenquimales mediante CD73, CD90, CD105 y el marcador de células troncales hematopoyéticas CD45 en el paso 1 (P1) y el paso 4 (P4), usando citometría de flujo y técnicas de inmunocitoquímica. Las células epiteliales respiratorias expresaron los marcadores de células troncales mesenquimales en P1 y mantuvieron la expresión de estos marcadores hasta P4. No hubo diferencias significativas en el uso de ambas técnicas al comparar los valores de los marcadores de células troncales mesenquimales expresadas desde P1 a P4. Este estudio indica que las células epiteliales respiratorias derivadas de la concha nasal retienen algunas de las propiedades de células troncales mesenquimales, incluso después de pases seriados. Ambos métodos de inmunofenotipificación son comparables.
Subject(s)
Humans , Biomarkers/metabolism , Epithelial Cells/cytology , Nasal Mucosa/cytology , Turbinates/cytology , Cell Culture Techniques , Flow Cytometry , Immunohistochemistry , Mesenchymal Stem Cells/cytology , Phenotype , Tissue EngineeringABSTRACT
Objective To assess the value of tumor marker immunostaining-FISH (TM-iFISH) technology on concen?trating and enumeration of tumor cells in CSF of lung cancer patients with leptomeningeal metastasis(LM). Methods Six cases of non-small cell lung cancer with leptomeningeal metastasis, which were diagnosed by CSF cytology or enhanced MRI scan, were selected. A total of 20 mL of CSF was collected in each case. TM-iFISH technology was employed to concen?trate and quantify circulating tumor cells in 7.5 mL CSF samples in each case while CSF cytology used 10 mL CSF samples in each case;Finally, the rest 2.5 mL CSF in each case was used for biochemistry assay. Results Ten CSF samples from 6 patients with non-small lung cancer with LM were assayed and tumor cells numbers ranging between 3 and 1 823 every 7.5 mL were found in 7 samples. On the other hand, CSF cytology examination only revealed tumor cells in 3 cases. Using CSF biochemical assay, higher than normal of protein level was found in 9 cases. TM-iFISH technology was employed again in 3 cases of patients who received treatment. Tumor cell count in CSF reduced in 2 out of the 3 cases. Conclusion TM-iFISH technology is a new method for detection and enumeration of tumor cells in the CSF in non-small cell lung cancer patients with leptomeningeal metastasis. This technology present diagnosis and curative values in lung cancer patients with leptomen?ingeal metastasis.
ABSTRACT
The present study was aimed to evaluate the influence of glutaraldehyde (GA) concentration on multiple electron microscopic (EM) immunostaining using pre-embedding peroxidase and post-embedding immunogold method. Influence of various concentrations of GA included in the fixative on immuoreactivity was assessed in the multiple immunostaining using antisera against anti-transient receptor potential vanilloid 1 (TRPV1) for peroxidase staining and anti-GABA for immunogold labeling in the rat trigeminal caudal nucleus. Anti-TRPV1 antiserum had specificity in pre-embedding peroxidase staining when tissues were fixed with fixative containing paraformaldehyde (PFA) alone. Immunoreactivity for TRPV1 was specific in tissues fixed with fixative containing 0.5% GA at both perfusion and postfixation steps, though the immunoreactivity was weaker than in tissues fixed with fixative containing PFA alone. Tissues fixed with fixative containing 0.5% GA at the perfusion and postfixation steps showed specific immunogold staining for GABA. The results of the present study indicate that GA concentration is critical for immunoreactivity to antigens such as TRPV1 and GABA. This study also suggests that the appropriate GA concentration is 0.5% for multiple immunostaining with peroxidase labeling for TRPV1 and immunogold labeling for GABA.
Subject(s)
Animals , Rats , gamma-Aminobutyric Acid , Glutaral , Immune Sera , Microscopy, Electron , Perfusion , Peroxidase , Sensitivity and Specificity , Trigeminal Caudal NucleusABSTRACT
Este estudio se llevó a cabo para examinar el patrón de inmunoexpresión simultánea de p16INK4a/Ki-67y establecer su posible utilidad clínica para la detección precoz del cáncer de cuello uterino. Las muestras celulares de cuello uterino fueron seleccionadas de la pesquisa de rutina de cáncer cervical. La detección inmunocitoquímica de p16INK4a/Ki-67se realizó con el kit de trabajo CINtec® Plus. Todos los casos tenían una prueba de virus papiloma humano (VPH). Ciento quince muestras citológicas fueron incluidas: 11(9,6%) fueron negativas para lesión intraepitelial o malignidad (NILM), 32(27,8%) presentaron células escamosas con atipias de significado indeterminado (ASC-US), 62(53,9%) mostraron lesión intraepitelial escamosa de bajo grado (LSIL) y 10(8,7%) lesión intraepitelial escamosa de alto grado (HSIL). La prevalencia general de infección por VPH fue de 81,7% (94/115). En 42 casos (45,0%) se identificaron los siguientes genotipos específicos de VPH: VPH16 (26,2%), VPH51 (21,4%), VPH52 (14,3%) y el genotipo VPH66 (7,1%). De 115 muestras celulares, 42(36,5%) fueron positivas para la tinción dual de p16INK4a/Ki-67, siendo ésta muy frecuente en las muestras citológicas con HSIL (70,0%), disminuyendo en las LSIL (44,0%) y existiendo inmunopositividad en una minoría de ASC-US (25,0%). Ningún caso NILM mostró inmunopositividad para p1(6INK4a)/Ki-67 (p<0,001). En 40/115 casos (34,8%) hubo positividad tanto para infección por VPH oncogénico como para la tinción dual p16INK4a/Ki-67, incluyendo 6/32 (18,8%) con ASC-US, 26/62 (42,0%) con LSIL and 8/10 (80,0%) con HSIL. Esta metodología podría ser utilizada para detectar lesiones en cuello uterino que aún no han sido diagnosticadas o han pasado inadvertidas.
We aimed to explore the expression pattern of p16INK4a/Ki-67 immunocytochemical dual-staining and to establish the potential clinical utility for early detection of cervical lesions. Liquid-based cytologies of cervical specimens of cervical cancer screening were processed for p16INK4a/Ki-67 immunocytochemical dual-staining using the CINtec® Plus Kit. HPV testing was performed with the INNO-LiPA HPV genotyping Extra Reverse Hybridization Line Probe Assay kit. One hundred and fifteen cervical cytologies were analyzed with the following results: 11(9.6%) were negative for intraepithelial lesions or malignancy (NILM); 32(27.8%) presented atypical squamous cells of undetermined significance (ASC-US); 62(53.9%) exhibited low grade squamous intraepithelial lesions (LSIL) and 10(8.7%) showed high grade squamous intraepithelial lesions (HSIL). No cases of cervical cancer were detected. The overall prevalence of DNA HPV detection was 81.7% (94/115). The following specific HPV genotypes were identified in 42 (45.0%) cases: HPV16 (26.2%), HPV51 (21.4%), HPV52 (14.3%) and HPV66 (7.1%). Viral sequences of an unknown single HPV were detected in 23.8%of the cases. A total of 42/115 (36.5%) were p16INK4a/Ki-67 dual-staining-positive, being more frequent in HSIL (70.0%), decreasing in LSIL (44.0%), detected in a minority of ASC-US (25.0%) and negative in NILM cases (p<0.001). 40/115 cases (34.8%) were positive for both oncogenic HPV and p16INK4a/Ki-67 dual-staining, including 6/32 (18.8%) ASC-US, 26/62 (42.0%) LSIL and 8/10 (80.0%) HSIL, which represent a strong association between positivity for HPV, p16INK4a/Ki-67 staining and severe cytological abnormalities (p<0.001). This methodology could be used to detect unnoticed cervical lesions.
Subject(s)
Adolescent , Adult , Female , Humans , Middle Aged , Young Adult , Papillomavirus Infections/pathology , Uterine Cervical Dysplasia/pathology , Uterine Cervical Dysplasia/virology , /analysis , /biosynthesis , Genotype , Immunohistochemistry , /analysis , /biosynthesis , Papillomaviridae , Staining and Labeling , Uterine Cervical Dysplasia/chemistry , Uterine Cervical Dysplasia/metabolismABSTRACT
BACKGROUND: Multiple myeloma (MM) is a heterogeneous and ultimately fatal disease. Risk stratification using prognostic biomarkers is crucial to individualize treatments. We sought to investigate the role of CD99, a transmembrane protein highly expressed in many hematopoietic cells including subpopulations of normal and neoplastic plasma cells, for MM risk stratification. METHODS: CD99 expression was measured in paraffin samples of bone marrow and extramedullary biopsies of 170 patients with MM. Patients were divided into those with high score (moderately and strongly positive) and low score (negative and weakly positive), with all staining being cytoplasmic and/or membranous. RESULTS: High anti-CD99 immunostaining was observed in 72 of 136 (52.9%) bone marrow biopsies and 24 of 87 (27.6%) extramedullary biopsies in MM. High CD99 expression of extramedullary specimens was associated with significantly longer overall survival (OS; p=.016). High CD99 expression of extramedullary specimens was also associated with better prognosis in the nonautologous stem cell transplantation group of MM patients (p=.044). In multivariate analysis, International Staging System stage was an independent prognostic factor, whereas CD99 expression was no longer statistically significant. CONCLUSIONS: Expression of CD99 in extramedullary specimens was correlated with longer OS, suggesting that CD99 may be a helpful immunohistochemical marker for risk stratification.
Subject(s)
Humans , Biomarkers , Biopsy , Bone Marrow , Cytoplasm , Multiple Myeloma , Multivariate Analysis , Paraffin , Plasma Cells , Prognosis , Stem Cell TransplantationABSTRACT
Aim To establish a highly purified,active and prac-tical extract and primitive culture method for rat brain microvas-cular endothelial cells ( BMECs) for providing materials for con-struction in vitro blood-brain barrier ( BBB) model. Methods Cerebral cortex of 1-2 week SD rats was collected,and successive digestion with typeⅡcollagenase and collagenase/dispase, sieve filtration and then twice gradient centrifugation in 20% BSA and 44% Percoll condition were used to obtain brain microvascular section. After that brain microvascular section was seeded in cul-ture bottle and then primarily cultured. Inverted microscope and factor-VIII relative antigen immunostaining methods were used for cellular morphological observation and identification. Results BMECs climbed out the vessel segment and proliferated with ad-herence after 2 h in vitro culture,and they became typical peb-bles structure after further 3-4 d culture. Morever, factor-VIII relative antigen immunostaining identified that expression for the endothelial cells was positive,cytoplasm was brown and positive cells account for more than 99%. Conclusion Rat BMECs with high purity could be extracted and cultured by using the above methods,and it has great potential for construction in vitro BBB model and in depth studies on biological characteristics and func-tions of BMECs.