Spelling suggestions: "subject:"immunohistochemistry"" "subject:"mmunohistochemistry""
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Faster than light (microscopy): superiority of digital pathology over microscopy for assessment of immunohistochemistryClarke, E., Doherty, D., Randell, Rebecca, Grek, J., Thomas, R., Ruddle, R.A., Treanor, D. 15 June 2023 (has links)
Yes / Digital pathology offers the potential for significant benefits in diagnostic pathology, but currently the efficiency of slide viewing is a barrier to adoption. We hypothesised that presenting digital slides for simultaneous viewing of multiple sections of tissue for comparison, as in those with immunohistochemical panels, would allow pathologists to review cases more quickly.
Novel software was developed to view synchronised parallel tissue sections on a digital pathology workstation. Sixteen histopathologists reviewed three liver biopsy cases including an immunohistochemical panel using the digital microscope, and three different liver biopsy cases including an immunohistochemical panel using the light microscope. The order of cases and interface was fully counterbalanced. Time to diagnosis was recorded and mean times are presented as data approximated to a normalised distribution.
Mean time to diagnosis was 4 min 3 s using the digital microscope and 5 min 24 s using the light microscope, saving 1 min 21 s (95% CI 16 s to 2 min 26 s; p=0.02), using the digital microscope. Overall normalised mean time to diagnosis was 85% on the digital pathology workstation compared with 115% on the microscope, a relative reduction of 26%.
With appropriate interface design, it is quicker to review immunohistochemical slides using a digital microscope than the conventional light microscope, without incurring any major diagnostic errors. As digital pathology becomes more integrated with routine clinical workflow and pathologists increase their experience of the technology, it is anticipated that other tasks will also become more time-efficient.
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Changes in the Murine Nigrostriatal Pathway Following Pyrethroid and Organophosphate Insecticide Exposure: An Immunohistochemical StudyPittman, Julian Thomas 01 October 2002 (has links)
Parkinson's disease (PD) is a debilitating motor disorder that primarily afflicts older individuals (> 50yrs). Although its cause is unknown, many factors are thought to contribute to the disease. There is growing epidemiological evidence supporting a link between pesticide exposure and PD. The present immunohistochemical study was undertaken to characterize the role of insecticide exposure in the etiology of idiopathic PD. The insecticides selected for study were the pyrethroid permethrin (PE) and the organophosphate chlorpyrifos (CP), both of which possess properties that could damage or disrupt the nigrostriatal pathway, which is the principal neurodegenerative target in PD. The present study examined possible alteration of the amount of dopamine re-uptake transporter protein (DAT), within the striatum of the C57BL/6 mouse, using DAT antibodies, following low (0.8, 1.5 & 3.0 mg/kg) and high (200 mg/kg) doses of PE, respectively. Possible nigrostriatal terminal degeneration was examined using antibodies to tyrosine hydroxylase (TH), the rate-limiting enzyme in dopamine synthesis, following treatment with 50 mg/kg of CP alone or in combination with the high dose of PE. For both the high dose of PE alone and for the combined PE/CP treatment, glial fibrillary acidic protein (GFAP) antibodies were used to examine the possibility of non-degenerative tissue injury. Groups of matched treated/vehicle-control mice received three IP injections of the insecticide/dose of interest over a 2-week period. Counts of immunoreactive (IR) neuropil in the dorsolateral striatum were made from four pre-selected fields per striatal tissue section. Counts were compared between matched sections, processed on the same slide, from a treated mouse and its vehicle control. A mean difference score, across slides, for each treated/vehicle control pair was determined. All low dose PE groups showed a trend of decreased DAT IR neuropil, but only the 3.0mg/kg group showed a statistically significant reduction (p<.0078). The 200 mg/kg PE group showed a trend toward reduced TH IR neuropil that was not statistically significant, but a significant increase in GFAP IR (p = .048) was observed. No significant change in TH IR neuropil was observed for CP (50mg/kg) alone. A significant increase was observed for GFAP IR neuropil for the PE/CP (200/50 mg/kg) combination dose (p = .033). The combined insecticide treatment failed, however, to produce a significant change in TH IR within the striatum, compared to vehicle controls. These data suggest that the significant increases in GFAP IR neuropil, in the striatum, reflect some form of tissue insult, following exposure to a high dose of PE, or PE/CP in combination, that is insufficient to induce degeneration of dopaminergic terminals within the temporal interval investigated. Although such damage may be sufficient to account for previously reported decreases in maximal dopamine uptake observed with high doses of these compounds, the DAT IR data appear to indicate that this damage is unlikely to be a change in the amount of DAT in these high dose conditions. The decreases in striatal DAT IR neuropil observed for low doses of PE suggest an alteration in the normal integrity of the nigrostriatal pathway and in the route by which environmental toxins may enter dopaminergic neurons. / Master of Science
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Immunohistochemical characterization of neuronal cilia in the rat central nervous system.Hughes, Rhome 05 1900 (has links)
An anti-G"11 antibody was used to label neuronal cilia throughout the rat central nervous system. Immunoreactive cilia were observed in every examined region of the rat CNS, but not in monkey or mouse tissue. Antibodies to G"q and G"q/11 failed to label cilia. Immunoreactive cilia were observed as early as postnatal day 0 in spinal tissue, and postnatal day 3 in hypothalamic tissue. There was a statistically significant negative correlation between a region's mean cilium length and that region's distance to the nearest ventricle; regions nearest ventricles were those with the longest cilia. This correlation suggests neuronal cilia may function as chemosensors, detecting substances as they move out from the cerebrospinal fluid and into the extracellular space of the brain.
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The demonstration of estrogen receptors in various tumours: a study using immunohistochemistry and in situ hybridisation.Henwood, Anthony F January 2004 (has links)
In order to study the incidence of Estrogen Receptors (ER) in breast carcinoma, lung carcinoma and melanoma, an in situ hybridisation technique for ER mRNA (ER mRNA-ISH) was developed. Various technical aspects of the procedure including tissue fixation, hybridisation conditions, and demonstration technique were investigated in order to obtain an optimum technique for routine use. ISH results were compared with ER immunohistochemistry using the monoclonal antibodies ER1D5 and D5. Commercially available biotin labelled antisense oligonucleotides to ER, Poly A (total mRNA), and sense chromogranin (negative control) were applied to frozen and formalin-fixed paraffin sections of breast carcinomas. For frozen sections, various fixatives including formalin, alcohol, Schoobridge, Zamboni's and acetic- alcohol were compared. A direct streptavidin- eroxidase and an indirect demonstration method using anti-biotin were also compared. The effect of differing formamide concentrations and post hybridisation stringency washings were analysed. An optimised ISH technique was then applied to frozen sections of 21 cases of breast carcinoma and 11 cases of lung carcinoma. Results were compared to H222 staining on adjacent sections. The ISH technique was also optimised for use on formalin-fixed, paraffin-embedded sections of 28 breast carcinomas and 17 melanomas. The results were compared with ER1D5 and D5 immunohistochemistry done on adjacent sections. The occurrence of endogenous biotin was also studied on a range of normal tissues. Consistent ISH results were obtained when formamide was omitted from the hybridisation cocktail, high stringency post hybridisation washes were discarded, room temperature hybridisations and an indirect demonstration method were used. Fixation of frozen sections in acetic/ethanol gave more consistent results with good morphology and resulted in positive nucleolar staining in 90% of breast and 45% of lung carcinomas. Positive nucleolar staining was also present in frozen sections of one metastatic melanoma. In formalin fixed paraffin sections, acid hydrolysis and pronase treatment were required prior to ISH. Cytoplasmic and/or nucleolar ER mRNA-ISH staining was seen in 87% of breast carcinoma and 97% of melanoma studied. ER1D5 was present in 54% of breast carcinomas but was absent in all melanomas. D5, on the other hand, was found in 88% of the melanomas. In conclusion, ER mRNA-ISH can be successfully done on acetic/alcohol fixed frozen sections and formalin fixed paraffin sections. Formamide, high stringency washes and elevated hybridisation temperatures are detrimental to a successful ISH reaction and an indirect demonstration method (using anti-biotin) is preferred. Unfortunately, endogenous biotin can cause false positive ISH reactions and needs to be considered during interpretation. Results show that the localisation of ER mRNA in the nucleolus is specific. Both ER mRNA-ISH and ER immunohistochemistry indicate that melanomas and some lung carcinomas contain a receptor possibly similar to that in breast carcinomas. / Thesis (M.Sc.)--Department of Anatomical Sciences, 2004.
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The demonstration of estrogen receptors in various tumours: a study using immunohistochemistry and in situ hybridisation.Henwood, Anthony F January 2004 (has links)
In order to study the incidence of Estrogen Receptors (ER) in breast carcinoma, lung carcinoma and melanoma, an in situ hybridisation technique for ER mRNA (ER mRNA-ISH) was developed. Various technical aspects of the procedure including tissue fixation, hybridisation conditions, and demonstration technique were investigated in order to obtain an optimum technique for routine use. ISH results were compared with ER immunohistochemistry using the monoclonal antibodies ER1D5 and D5. Commercially available biotin labelled antisense oligonucleotides to ER, Poly A (total mRNA), and sense chromogranin (negative control) were applied to frozen and formalin-fixed paraffin sections of breast carcinomas. For frozen sections, various fixatives including formalin, alcohol, Schoobridge, Zamboni's and acetic- alcohol were compared. A direct streptavidin- eroxidase and an indirect demonstration method using anti-biotin were also compared. The effect of differing formamide concentrations and post hybridisation stringency washings were analysed. An optimised ISH technique was then applied to frozen sections of 21 cases of breast carcinoma and 11 cases of lung carcinoma. Results were compared to H222 staining on adjacent sections. The ISH technique was also optimised for use on formalin-fixed, paraffin-embedded sections of 28 breast carcinomas and 17 melanomas. The results were compared with ER1D5 and D5 immunohistochemistry done on adjacent sections. The occurrence of endogenous biotin was also studied on a range of normal tissues. Consistent ISH results were obtained when formamide was omitted from the hybridisation cocktail, high stringency post hybridisation washes were discarded, room temperature hybridisations and an indirect demonstration method were used. Fixation of frozen sections in acetic/ethanol gave more consistent results with good morphology and resulted in positive nucleolar staining in 90% of breast and 45% of lung carcinomas. Positive nucleolar staining was also present in frozen sections of one metastatic melanoma. In formalin fixed paraffin sections, acid hydrolysis and pronase treatment were required prior to ISH. Cytoplasmic and/or nucleolar ER mRNA-ISH staining was seen in 87% of breast carcinoma and 97% of melanoma studied. ER1D5 was present in 54% of breast carcinomas but was absent in all melanomas. D5, on the other hand, was found in 88% of the melanomas. In conclusion, ER mRNA-ISH can be successfully done on acetic/alcohol fixed frozen sections and formalin fixed paraffin sections. Formamide, high stringency washes and elevated hybridisation temperatures are detrimental to a successful ISH reaction and an indirect demonstration method (using anti-biotin) is preferred. Unfortunately, endogenous biotin can cause false positive ISH reactions and needs to be considered during interpretation. Results show that the localisation of ER mRNA in the nucleolus is specific. Both ER mRNA-ISH and ER immunohistochemistry indicate that melanomas and some lung carcinomas contain a receptor possibly similar to that in breast carcinomas. / Thesis (M.Sc.)--Department of Anatomical Sciences, 2004.
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Immune escape mechanisms in EBV-associated nasal NK/T-Cell lymphomaShen, Lijun., 沈立軍. January 2002 (has links)
published_or_final_version / Pathology / Doctoral / Doctor of Philosophy
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Tumor de Leydig simulando una neoplasia germinalGamboa Acuña, Brenda Adriana, Guillén Zambranoa, Rayza, Lizzetti Mendozaa, Grecia 22 June 2016 (has links)
Main findings
A case is presented of a Leydig cell neoplasm in a 25 year-old male patient with no classic risk factors with an atypical outcome. The tumour mass was histologically analysed and was found to have features compatible with a germ cell neoplasm. A right orchiectomy was performed, followed by chemotherapy. After treatment, pulmonary metastasis, lymph nodes, and peri-hepatic hydronephrosis were found. The patient died two months after his last hospital admission.
Case hypothesis
Leydig cell tumours account for 1% to 3% of all testicular tumours. They occur at any age, especially in children, and between the third and sixth decade of life. Around 90% are benign, and 10% are malignant; these latter usually occurring between 50 and 60 years old, and are associated with sizes larger than 5 cm and gynecomastia. Finally, it is difficult to predict the development of the disease based on histopathological observations.
Promising future implications
Although non-germ cell tumours are rare, it is important to consider them in the differential diagnosis of testicular tumours, particularly in those of non-seminoma origin. Immunohistochemistry is useful for the differentiation of testicular tumours in those cases when conventional histology shows no conclusive findings. / Hallazgos principales
Reportamos un caso de neoplasia de células de Leydig en un paciente varón de 25 años, sin factores de riesgo clásicos con evolución tórpida. Se analizó la histopatología de la masa tumoral y se encontró malignidad por lo que se decide realizar orquiectomía derecha, seguida de quimioterapia. Luego del tratamiento se halla metástasis pulmonar, adenopatías perihepáticas e hidronefrosis, falleciendo 2 meses después de su último ingreso hospitalario.
Hipótesis del caso
El tumor de células de Leydig representa entre el 1-3% de todos los tumores testiculares. Se presentan a cualquier edad; sobre todo en la infancia y en la 3.a-6.a década de la vida. Aproximadamente el 90% son de curso benigno y el 10% son malignos, presentándose sobre todo entre la 5.a y la 6.a década de la vida, y están asociados con un tamaño > 5 cm y ginecomastia. Finalmente, es difícil predecir el comportamiento en términos histológicos.
Repercusiones a futuro
A pesar que los tumores de células no germinales son poco frecuentes, es importante considerarlos como diagnóstico para brindar el tratamiento óptimo y evitar resultados desafortunados.
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Vliv bilirubinu na progresi nespecifických střevních zánětů. / Bilirubin influence on the progression of inflammatory bowel disease.Patková, Anna January 2014 (has links)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Bilirubin influence on the progression of inflammatory bowel disease Diploma thesis Anna Patková Supervisor: doc. PharmDr. Petr Nachtigal, Ph.D. Background: Inflammatory bowel diseases, including Crohn's disease and ulcerative colitis, are chronic inflammatory disorders of the gut caused by an interaction of genetic and environmental factors. It is thought that tissue damage is also partly caused by an oxidative stress. Heme oxygenase-1 and bilirubin are strong antioxidants and both of them provide an anti-inflammatory effect in various tissues. The aim of this diploma thesis was to detect changes of expression of HO-I in the large intestine of normobilirubinemic and hyperbilirubinemic rats after the induction of acute or chronic experimental colitis. Methods: We used Gunn rats with hereditary defect of UDP-glucuronyltransferase, which causes hyperbilirubinemia. The control group of animals was made up of heterozygous littermates of the Gunn rats, which have normal serum bilirubin levels. All animals were treated by dextran sulfate sodium in order to induce an experimental colitis. Rats were divided into two groups. Each of them contained hyperbilirubinemic and normobilirubinemic...
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Aspects of Gene Expression Profiling in Disease and HealthBergman, Julia January 2017 (has links)
The aim of this thesis is to in various ways explore protein expression in human normal tissue and in cancer and to apply that knowledge in biomarker discovery. In Paper I the prognostic significance of RNA-binding motif protein 3 (RBM3) is explored in malignant melanoma. To further evaluate the prognostic significance of RBM3 expression was assessed in 226 incident cases of malignant melanoma from the prospective populationbased cohort study Malmö Diet and Cancer Study using tissue microarray technique (TMA). RBM3 was shown to be down regulated in metastatic melanoma and high nuclear expression in the primary tumor was an independent marker of prolonged over all survival. As a tool to facilitate clinical biomarker studies the Human Protein Atlas has created a tissue dictionary as an introduction to human histology and histopathology. In Paper II this work is introduced. A cancer diagnosis can be a complex process with difficulties of establishing tumor type in localized disease or organ of origin in generalized disease. Immunohistochemically assisted diagnosis of cancer is common practice among pathologists where its application combined with known protein expression profiles of different cancer types, can strengthen or help dismiss a suspected diagnosis. In Paper III the diagnostic performance of 27 commonly used antibodies are tested in a predominantly metastatic, multicancer cohort using TMA technique. Overall these 27 diagnostic markers showed a low sensitivity and specificity for its intended use, highlighting the need for novel, more specific markers. Breast, ovarian, endometrial and ovarian cancers affect predominantly women. Differential diagnostics between these cancer types can be challenging. In Paper IV an algorithm, based on six different IHC markers, to differentiate between these cancer types is presented. A new diagnostic marker for breast cancer, namely ZAG is also introduced. In Paper V the transcriptomic landscape of the adrenal gland is explored by combining a transcriptomic approach with a immunohistochemistry based proteomic approach. In the adrenal gland we were able to detect 253 genes with an elevated pattern of expression in the adrenal gland, as compared to 31 other normal human tissue types analyzed. This combination of a transcriptomic and immunohistochemical approach provides a foundation for a deeper understanding of the adrenal glands function and physiology.
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Expressão imuno-histoquímica de Bcl-2, Ki-67 e caspase-3 ativa em líquen plano oral e leucoplasia com diferentes graus de displasia / Immunohistochemical expression of Bcl-2, Ki-67 and active caspase-3 in oral lichen planus and leukoplakia with different degrees of dysplasiaPigatti, Fernanda Mombrini 26 August 2011 (has links)
O líquen plano oral (LPO) é uma doença inflamatória crônica de causa desconhecida, e seu potencial de malignização é um tema bastante controverso. Diversos estudos têm sugerido que pacientes portadores de líquen plano apresentam um maior risco de desenvolver câncer. Contudo, muitos autores acreditam que não haja dados suficientes que provem uma associação entre líquen plano e câncer. Para esses autores, a maioria dos casos que sofreram transformação maligna é decorrente de falhas no diagnóstico inicial da lesão. Portanto, objetivou-se avaliar a expressão imuno-histoquímica das proteínas relacionadas à apoptose e proliferação celular, respectivamente caspase-3 ativa, Bcl-2 e Ki-67 no LPO e em displasias epiteliais na tentativa de explicar a polêmica em relação ao potencial de transformação maligna do LPO e enfatizar a importância de um acompanhamento de longo prazo dos pacientes com esta doença. Com esse propósito, foram selecionadas 14 amostras de LPO, 14 amostras de leucoplasia com displasia epitelial, além de 09 amostras de mucosa bucal normal como controle. A avaliação da expressão de caspase-3 ativa, Bcl-2 e Ki-67 foi conduzida de acordo com a técnica da imunoperoxidase. A contagem das células imunomarcadas nas amostras de LPO, de leucoplasia com displasia epitelial e de mucosa bucal normal resultou em número destas células por mm2 nas diferentes regiões (camada basal, camada suprabasal e infiltrado inflamatório) para cada imunomarcador. A expressão de Bcl-2 em células epiteliais, da camada basal, ocorreu mais frequentemente no grupo da leucoplasia com displasia epitelial, com diferença estatística entre o grupo do LPO e o grupo controle. Verificou-se também, uma alta expressão da proteína Bcl-2 em células inflamatórias de lesões de LPO e de leucoplasia com displasia epitelial. A expressão do marcador Ki-67 foi superior em todos os níveis teciduais analisados nas lesões de LPO e de leucoplasia com displasia epitelial quando comparados com o grupo controle. Concluiu-se que a expressão mais elevada das proteínas Bcl-2 e Ki-67 nos casos de LPO e de leucoplasia com displasia epitelial, podem revelar a possibilidade da presença de alterações moleculares morfologicamente imperceptíveis. / The oral lichen planus (OLP) is a chronic inflammatory disease of unknown cause, and its malignant potential is a very controversial issue. Several studies have suggested that patients with lichen planus have a higher risk of developing cancer. However, many authors believe that there is insufficient evidence to prove an association between lichen planus and cancer. For these authors, most of the cases that had undergone malignant transformation is due to flaws in the lesion initial diagnosis. Therefore, the aim was to evaluate the immunohistochemical expression of proteins related to apoptosis and cell proliferation, respectively active caspase-3, Bcl-2 and Ki-67 in epithelial dysplasia and LPO in an attempt to explain the controversy regarding the potential malignant transformation of OLP and emphasize the importance of a long-term monitoring of patients with this disease. For this purpose, we selected 14 samples of OLP, 14 samples of leukoplakia with epithelial dysplasia, and 09 samples of normal oral mucosa as controls. The evaluation of the expression of active caspase-3, Bcl-2 and Ki-67 was conducted in accordance with the immunoperoxidase technique. The immunostained cells counting in samples of OLP, epithelial dysplasia in leukoplakia and normal oral mucosa resulted in a number of these cells per mm2 in the different regions (basal layer, suprabasal layer and inflammatory infiltrate) for each immunostained. The expression of Bcl-2 in epithelial cells, the basal layer, occurred more frequently in the group of leukoplakia with epithelial dysplasia, with statistical difference between the group of OLP and the control group. There was a high expression of Bcl-2 protein in inflammatory cells in OLP lesions and leukoplakia with epithelial dysplasia. The expression of the marker Ki-67 was superior in all analyzed tissue levels in OLP lesions and leukoplakia with epithelial dysplasia when compared to the control group. It was concluded that the highest expression of Bcl-2 protein and Ki-67 in cases of OLP and leukoplakia with epithelial dysplasia, can reveal the possible presence of molecular changes morphologically imperceptible.
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