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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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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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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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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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Immunohistochemical analysis of the expression of selected signaling molecules in rat brain: the effect of morphine withdrawal
Přítulová, Eliška ; Novotný, Jiří (advisor) ; Mrózková, Petra (referee)
Morphine is one of the most commonly used analgesics for pain, but its clinical use may be accompanied by the development of tolerance and dependence. Abuse of morphine can then lead to the development of severe withdrawal symptoms. Given the knowledge gathered so far, morphine addiction research often examines the impact on areas of the brain involved in the reward system. The main goal of this work was to investigate the effect of long-term administration of morphine and morphine withdrawal on certain signaling molecules that are related to the molecular action of opioids in selected areas of the rat brain. We focused on the cAMP response element-binding protein (CREB) and its active phosphorylated form (pCREB), as well as on G-protein signaling regulator RGS4. Our results indicated that morphine administration may cause a decrease in CREB expression in the basolateral amygdala in morphine-affected rats. We also found a reduction in CREB phosphorylation in the CA1 region of the hippocampus, possibly due to morphine withdrawal for three months. In this study, we did not observe any statistically significant changes in RGS4 expression in the prefrontal cortex and hippocampus due to morphine administration and subsequent morphine withdrawal. Key words: morphine, dependence, withdrawal, rat brain,...
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Bilirubin influence on the progression of inflammatory bowel disease.
Patková, Anna ; Nachtigal, Petr (advisor) ; Hronek, Miloslav (referee)
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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Analysis of embryotoxic effect of hydrocortisone using chick embryotoxicity screening test (CHEST).
Janíková, Michaela ; Peterka, Miroslav (advisor) ; Hovořáková, Mária (referee)
Cleft lip is one of the most common human birth deffects. Its etiopathogenesis is multifactorial and many aspects of its occurrence remain unknown in the fields of both genetics and teratology. One of the set of known negative external factors causing cleft lip is chemical hydrocortisone. Its effect on cell proliferation is highly heterogeneous and depends on attributes of a specific cell population. In this work we studied the cleft beak origin after the hydrocortisone treatment on the basis of Chick Embryotoxicity Screening Test (CHEST). Our main aim was to detect cell cycle changes in the chick frontonasal process after hydrocortisone injection via flow cytometry analysis. Hydrocortisone caused S phase arrest within a minor subpopulation of highly granular cells with specific cell cycle. This sensitive subpopulation was localized in the areas of previously defined proliferative centers within the frontonasal process using immunohistochemistry of frozen sections. Quantitative analysis of cells in these areas revealed significant decrease of M phase portion in the hydrocortisone treated samples in comparison with the control samples. The TUNEL staining of histological sections was used to determine the apoptotic rate in the frontonasal process. The comparison between the control and the...
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Morphological and Immunohistochemical Analysis of Melanocytic Lesions
Důra, Miroslav ; Dundr, Pavel (advisor) ; Mandys, Václav (referee) ; Pock, Lumír (referee)
This doctoral thesis deals with morphological and immunohistochemical analysis of melanocytic lesions. Melanoma is the most dangerous malignant skin tumor with its potential occurrence outside the skin. Its early diagnosis is the basic aspect of a favorable disease prognosis. Distant metastases development occurring several years after the initial diagnosis is a typical feature of melanoma. Histopathological differential diagnostics of melanocytic lesions is extremely difficult because of the diversity of melanocytic lesions. Contemporary research is focused on the identification of new surrogate markers based on immunohistochemistry or molecular genetics, which would be enable to estimate the behavior of a certain lesion and clarify the gray zone of melanocytic lesions of uncertain biological behavior. The doctoral thesis is focused on the immunohistochemical analysis of GLUT-1 protein expression in 400 cases of benign and malignant melanocytic lesions. Significant expression of this marker was identified in the group of melanomas, it was not identified in melanocytic nevi. Expression of GLUT-1 corresponded to the worse prognosis of the disease. The doctoral thesis also deals with the issue of TILs. The analysis of TILs (tumor infiltrating lymphocytes) in 213 melanomas compared the prognostic...
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