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Histopathological predictors of clinical course severity and development of surgical complications in children with inflammatory bowel diseases
Fabián, Ondřej ; Zámečník, Josef (advisor) ; Daum, Ondřej (referee) ; Karásková, Eva (referee)
The aim of the presented thesis is to summarize a benefit and limitations of histopathological assessment of bioptic samples for diagnostics and prediction of complicated course of the disease in children with inflammatory bowel diseases (IBD). We would also like to outline a potential benefit of more sophisticated histopathological methods in these domains. In the first phase of our research, we aimed at correlation of microscopical disease activity in children with Crohn's disease (CD) with activity assessed endoscopically and clinically. We also examined a histopathology as a possible predictor of development of complications, since previous works on adult cohorts indicate only a limited predictive value of microscopy. Our study confirmed this premise also for pediatric patients, since histopathological findings correlated weakly with endoscopy, didn't correlate with clinical activity of the disease and did predict none of the defined complications. On the other hand, endoscopy appeared to be a reliable predictor of complicated course of the disease. In the next part of our research, we were searching for other, more precise methods of assessment of histopathological disease activity that could serve as predictors of complications. We focused at immunohistochemical assessment of tissue...
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Diagnostics of Helicobacter pylori using histological and immunohistochemical examination
BURDOVÁ, Anna
The main topic of this bachelor thesis is the bacterium Helicobacter pylori, its occurrence in the gastric mucosa, and the metod of its diagnosis in the laboratory using histological and immunohistochemical examination. In the theoretical part, I describe the bacterium itself, its morphology and mainly its diagnostics. There are many ways to detect the presence of this bacterium, most commonly divided into invasive tests, which include histological examination, cultivation and urease test, and non-invasive, where we can use serological and breathing tests. I also describe the anatomy and histological structure of the stomach in this part. Then I follow up on stomach pathology, where I focused on the most common diseases caused by Helicobacter pylori, acute and chronic gastritis and stomach tumors. The over-all histological examination is also included, its parts being receiving the sample to the department through fixation with examples of various fixation agents, to embedding, cutting and staining, where I also describe the types of staining that are used in the practical part. I also deal with the topic of immunohistochemistry. One of the goals of the practical part is to master the basic routine Hematoxylin eosin staining, special Giemsa staining and immunohistochemical examination of the antigen using the Anti-Helicobacter pylori polyclonal antibody. Another goal is to map the occurrence of Helicobacter pylori infection based on age groups, sex of examined patients and season. I set two research questions that I would like to answer. Results obtained in the year 2022 will be statistically processed and supplemented with photomicrographs of selected samples.
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Diagnostic of immunohistochemistry in pathology
HRDINOVÁ, Andrea
The bachelor thesis deals with the use of immunohistochemical methods in the diagnosis of neoplasms in pathology. Describes the use of antibody spectrum at the Department of Pathology, Český Krumlov Hospital, a.s. Immunohistochemical methods are nowadays an important method used in pathology for tumor typing and metastasis for the determination of primary tumors. They complement the standard staining methods for basic diagnostics such as Hematoxylin-Eosin and Van-Gieson, which are less specific, these methods mainly serve to differentiate the basic cellular components and tissue components of the preparation. Immunohistochemistry methods are based on the special antibody affinity principle and are divided into direct and indirect methods. For a direct method, an antibody labeled with, for example, an enzyme is used to detect the antigen and, upon binding, induces a color change observed under the microscope. Indirect method (two-stage, three-stage) is a more sensitive method, based on the principle of an unlabeled primary antibody to which a conjugated secondary antibody binds to an enzyme or biotin. In the three-stage, we use the affinity of avidin and biotin to form the avidin-biotin-enzyme complex (eg. peroxidase), which is shown and highlighted. In more detail, the thesis deals with the diagnosis of breast and sentinel lymph node tumors, both in the theoretical and practical part, and the incidence of these tumors since the introduction of the immunohistochemical method at the Department of Pathology of the Český Krumlov Hospital. Last but not least, a case report of individual interesting cases is described.
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Breast cancer immunohistochemistry
HYŽÍKOVÁ, Sabina
In my bachelor thesis I deal with imunohistochemical examination of breast cancer. I performed the research at the Department of pathology at the hospital in Písek. The thesis is divided into theoretical and practical part In the theoretical part I deal with anatomical structure of the breast, clinical symptoms, diagnostics together with imaging methods. Next risk factors, including genetic and hormonal aspects of breast cancer. Prognosis and also by prevention of tumor development and subsequent treatment of this disease. I summarized the general classification of tumors representing staging and grading. I also described pathological structures of the breast as benign and malignant lesions. In the practical part I deal with the description of the immunohistochemical method, I focused on how the method works and especially how the whole histological process looks like. I worked with a total amount of 218 biopsy samples of breast tissue, which were subsequently stained with this method and I determined 5 markers (oestrogen and progesterone receptors, HER-2/Neu, Ki67 and E-cadherin), which were used in their diagnosis. Based on the negativity of ER, PR and HER-2/Neu, I evaluated fourteen triple-negative lesions. Hormone receptors showed numerous positive results more than negative ones, while HER-2/Neu expression showed only slightly higher positivity in hundred and twenty two cases compared to the remaining ninety six negative cases. The results of the practical part of my bachelor thesis was finding, that the most prevalent histologic subtype of breast cancer is the invasive ductal carcinoma, which was represented in one hundred and ninety two cases, another type was invasive lobular carcinoma represented in twenty two cases and six ductal carcinoma in situ. The aim of my bachelor thesis was to present a brief description of the issue of breast cancer and point out the representation of the most frequently diagnosed subtypes as well as to describe the execution of a method that is typical for detecting breast cancer-associated receptors.
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Histological examination of the colon
FEJKOVÁ, Karolína
This bachelor´s thesis deals with histological examination of the colon, namely the laboratory diagnosis of colorectal cancer. Colorectal cancer is a highly relevant issue today not only in the Czech Republic, not least because of its continually growing incidence worldwide. Laboratory analysis of this disease is not only important from the aspect of diagnosis per se. Determining the type and invasive stage of the disease is also crucial for the therapeutic strategy in a given patient. Immunohistochemistry testing of the colon also aids in the diagnosis of Lynch syndrome, a hereditary disease with a high risk of colorectal cancer. The first part of the thesis is theoretical and aims to introduce not only the subject of colorectal cancer but also the anatomical, histological and pathological features of the colon. This section also describes the methods used to obtain tissue samples from the colon and to process these in a histopathology laboratory. This information regarding the histopathological processing of colon tissue is then applied and described in detail in the methodology section of the thesis. This describes the process of routine staining methods such as haematoxylin-eosin and Alcian Blue. It also details the manually performed immunohistochemical methods and related reagents, including the use of antibodies. The next section presents a statistical analysis of a sample of patients with colorectal cancer treated at the Jihlava Hospital between 2018 and 2019. These data confirm the generally accepted fact that there is a higher incidence of colorectal cancer in men and that this cancer predominantly affects patients aged between 60 to 70 years. Furthermore, this work shows that defects in the MMR proteins associated with Lynch syndrome are not detected in even one half of the patients tested.
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Immunohistochemical and histological correlation of melanoma lesions
ČAPKOVÁ, Denisa
The bachelor thesis deals with the problematic of melanoma lesions. This theme is actual because melanoma is the most serious type of skin cancer and its incidence is moving to the younger generation. Because melanomas occur primary due to ultraviolet light exposure, one of the best possible ways to reduce the risk of developing melanoma is to protect yourself from ultraviolet radiation by using the sunscreen, wearing clothing that covers arms and legs and avoid using tanning beds. The prognosis of early melanoma is usually very good, so the detection and treatement is very important. In the theoretical part the current knowledge of melanoma, the characteristic, the occurence, the clasification, the prevention and the current possibilities of treatment were summarized. In the practical part the Hematoxylin eosin staining and the immunohistochemical method was described. There is also comparation of positive HMB-45 marker between each histological groups. The HMB-45 marker is absolutly specific for melanocytic tumors and has great utility fot the pathologist in differentiation of tumors with uncertain origin. From 2012 to 2018 were imunohistologicaly examined at the Departement of Pathology Nemocnice Písek a.s. 61 samples. Out of total 61 samples were 58 HMB-45 positive. The thesis has been focused on the imunohistological profile of each histological group. The imunohistologichemical profile was proven in 92%.
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Lynch syndrome in patients with upper urinary tract urothelial carcinoma: One centre study
BUCHOVÁ, Karolína
Lynch syndrome (LS) is an inherited autosomal dominant (AD) disease with predisposition for cancer development in different organs (large intestine, uterus, upper urinary tract, etc.). Typically, young or middle age individuals are affected by cancer. Presented bachelor thesis summarizes the current knowledge about LS in patients with urothelial carcinoma (UC) of the upper urinary tract. We tried to determine the frequency of LS in patients with urothelial carcinoma of the upper urinary tract and design a suitable diagnostic algorithm how to identify suspect patients appropriate for further genetic testing. In the practical part of the thesis, we searched for all patients treated on Urology department Faculty Hospital in Pilsen for UC of the upper urinary tract in the time period I/2010 - XII/2018. All cases were re-evaluated. Immunohistochemical staining of MMR proteins (MLH1, PMS2, MSH2, MSH6) was performed in selected cases. Obtained data were evaluated statistically. We found 215 examinations/biopsies from 182 patients (58 UC of the ureter, 119 UC of the renal pelvis, 5 UC of both the ureter and the renal pelvis). 121 patients were examined by immunohistochemistry (44 UC of the ureter, 73 UC of the renal pelvis, 4 UC of both the ureter and the renal pelvis). Lost expression of some MMR protein was demonstrated in 9/121 examined cases. Definitive examination of peripheral blood for detection of germline mutation was performed in only two patients (2%). One patient (male, 71 years) has confirmed germline mutation of MSH6 gene from peripheral blood. The second patient (male, 73 years) is still waiting for a definitive confirmation of the diagnosis from peripheral blood (patient has a high suspicion for Lynch syndrome, he has personal history of colorectal carcinoma, deficiency of MSH6 protein expression was found in UC of upper urinary tract even in colorectal carcinoma). Based on here presented data, we recommend routine immunohistochemical staining of MMR proteins in all patients with UC of upper urinary tract, regardless of their age or medical history. Universal immunohistochemical screening in patients with UC of upper urinary tract is a good and yielding way how to identify suspicious patients for genetic testing of LS.
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Metodické přístupy k vyšetření lymfoidních tkání jeseterů
ŠÁLKOVÁ, Eva
We present material, data and results that were encountered in doctoral study and summarized in dissertation thesis. Our research was focused on methodical approaches to examination of lymphoid tissues in sturgeons. Sturgeons live on Earth for a long period of time. Well-developed immune system enabled sturgeons to face and deal with different attacks, and thus played an important and crucial role in sturgeon survival until present days. Sturgeons exhibit large genomes among fishes and are well known for their polyploidy origin. Acipenseriformes have a high tolerance for hybridization as well as for spontaneous doubling of chromosome sets. In their evolution, at least three independent polyploidization events have taken place, in the literature known as 3R hypothesis (three rounds hypothesis). Therefore, the present research focused on the haematopoietic and lymphoid tissues examination. Representatives of different ploidy level groups were encountered in the study: sterlet (Acipenser ruthenus) represented diploid (2n) group, Russian sturgeon (Acipenser gueldenstaedtii) was chosen as tetraploid group representative (4n), and finally shortnose sturgeon (Acipenser brevirostrum) served as hexaploid group representative (6n). Haematopoietic and lymphoid tissues were conventionally histologically processed, obtained sections were stained using haematoxylin and eosin. Advanced histological staining methods were applied when examining tissues to map and recognize particular cell types involved in haemato- and lymphopoiesis. Hassall's corpuscles were detected in thymi, particular in the thymic medulla; to our knowledge, it was the first description of Hassall's corpuscles in sturgeons. Subsequently, the thymus of juvenile sterlet (Acipenser ruthenus) was examined immunohistochemically. The wide range cytokeratin, vimentin, S-100 protein, leukocyte common antigen LCA (CD45) and CD3 were selected as the markers to map the thymic microenvironment. Commercially available human antibodies were applied on fish tissue, and wide range cytokeratin, vimentin, S-100 protein brought conclusive results and distinguished nature of different microenvironmental components. Contrarily, LCA and CD3 application was negative or inconclusive, and failed. Study confirmed the possibility of human antibody application on fish tissue, together with the need to generate fish and species specific antibodies, mostly when examining lymphoid tissues. Except for the histological and immunohistochemical examination, respectively, the lymphoid or immune system should be tested when using the determination of white blood cell parameters. Generally, the blood is more easily accessible (vessel puncture), and the determination of the total number of leukocytes and the differential counts represent important fish health and physiology status. Under the study, representatives of three different ploidy level groups were tested and white blood cell parameters were monitored monthly during a period of one year. Data suggested the significant effect of ploidy level on total number of leukocytes and morphological nuclear changes in granulocytes and lymphocytes. Annual variation in differential leukocyte counts depended on the species and influence of various external conditions rather than ploidy level. Examination of white blood cell parameters (total number of leukocyte and differential count), as well as histological and immunohistochemical examination, respectively, represent the main methodical approaches for lymphoid organs and immune system assessment in sturgeons. Methods should be applied in an ichthyological practise. Obtained results contribute to fish health status evaluation.
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Detection and characterization of macrophages in the tumors of viral and non-viral etiology
Dalewská, Natálie ; Tachezy, Ruth (advisor) ; Krulová, Magdaléna (referee)
Head and neck cancers are etiologically associated with smoking and alcohol consumption. Part of these tumors is induced by HPV and their incidence is increasing in the last decade. Patients with virally induced tumors have better prognosis even though they are usually diagnosed with tumors in advanced stage. One of the possible explanations may be better stimulation of the immune system by viral antigens. Macrophages are cells of the innate immune system which belong to professional phagocytes. They are called TAM upon infiltration to the tumor where they represent heterogeneous group of cells. Two main phenotypes are antitumor M1 and protumor M2 macrophages. TAMs are a major component of tumor microenvironment of many types of tumors, one of them are also head and neck cancers. In my thesis I focused on the immunohistochemical detection of M1 and M2 macrophages in the head and neck tumors of viral and non-viral etiology and at the same time RT-qPCR analyses of gene expression of macrophage-associated and/or immunosuppressive genes IDO1, ARG1, CD163, NOS2 a PTGS2 was performed. My data showed that HPV- negative tumors had higher number of M2 macrophages with typical markers CD163, ARG1 and PTGS2. It is known that patients with these tumors have worse prognosis of the disease. Due to high...
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Morphological, immunohistochemical, and molecular analysis of leiomyocellular tumors of the female reproductive system
Gregová, Mária ; Dundr, Pavel (advisor) ; Mandys, Václav (referee) ; Škarda, Jozef (referee)
Introduction Leiomyoma with bizarre nuclei (LBN) and cellular leiomyoma (CL) are rare variants of uterine smooth muscle tumors. In diagnostic practice, LBN can be mistaken for leiomyosarcoma (LMS), while CL may mimic low grade endometrial stromal sarcoma (LG ESS). Careful evaluation of morphological features is necessary when making the diagnosis; in some borderline cases, immunohistochemical and molecular examinations may help. Literature data on molecular genetic alterations in LBN and CL is limited, but some of these tumors appear to share certain aberrations with classical leiomyomas (UL) and LMS. Aims The aim of the work is to expand the knowledge about smooth muscle tumors of the uterus, especially LBN and CL, and perform a complex morphological, immunohistochemical (IHC), and molecular evaluation of their features. The individual goals include: 1) confirmation of the hypothesized benign behaviour of LBN, 2) morphological analysis of LBN, 3) more detailed clarification of LBN tumorigenesis with a focus on the FH gene, 4) clarification of CL tumorigenesis, 5) the use of IHC FH antibody as a screening method to identify FH gene mutations, 6) the use of morphological evaluation and results of IHC examination to facilitate differential diagnostic balance between benign and malignant smooth muscle...
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