Biomedical Papers-Olomouc最新文献

Backround: The goal of assisted reproduction is for a couple treated with IVF techniques to end the treatment by giving birth to a healthy baby. A neccessary presumption for success is the identification of the best embryo with high implantation and developmental potential. One option is to select an euploid embryo by invasive preimplantaion genetic testing for aneuploidy (PGT-A) or it is possible to select the best embryo by non-invasive time-lapse monitoring (TLM), specifically based on morphokinetic parameters and morphological markers that are able to identify an embryo with high developmental potential.

Materials and methods: The study involved a total of 1060 embryos (585 euploid and 475 aneuploid embryos after PGT-A) with good morphology from 329 patients in the period 01/2016-10/2021. All embryos were cultured in a time-lapse incubator, trophectoderm (TE) cells biopsies for PGT-A examination were performed on day 5 (D5) or day 6 (D6) of culture. During the study period, 225 frozen embryo transfers (FET) of one euploid embryo were performed. Based on the treatment outcome, the embryos were divided into 2 groups - euploid embryos, which led to the birth of a healthy child, and euploid embryos that did not show fetal heartbeat (FHB) after FET.

Results: Based on the statistical analysis of the embryos without implantation and the embryos with live birth, it is clear that the morphokinetic parameters t5 (time of division into 5 cells) and tSB (time of start of blastulation) are significantly different.

Conclusion: The results suggest that of the morphokinetic parameters tSB and t5 are predictive indicators for selecting an embryo with high developmental potential and with a high probability of achieving the birth of a healthy child.

Background: We report four pediatric subjects with Cushing's disease (CD) diagnosed in the Czech Republic. We focus on initial symptoms of Cushing's syndrome (CS) which can lead to early diagnosis, on typical symptoms of CS in children, their age and sex distribution, the mean length of symptoms prior to diagnosis, indication for examination, post-cure growth, sexual development and pituitary function in our four CD patients after transsphenoidal pituitary surgery (TSS). We describe the diagnostic process leading to confirmation of CD and we emphasize the biochemical and radiological diagnostic difficulties.

Conclusions: Pediatric CD has a number of features distinct from adult CD. Our retrospective analysis confirmed the presence of growth retardation and change in facial appearance with development of moon face as the first symptoms of CS. According to our observation, growth retardation is prior to development of moon face. The other typical symptoms frequently seen in pediatric patients are pseudo-precocious puberty in both sexes, hirsutism in pubertal girls due to excessive adrenal androgen secretion and pubertal delay. A corticotropin-releasing hormone (CRH) test and especially bilateral inferior petrosal sinus sampling for ACTH (BIPSS) contribute to confirming the diagnosis of CD and excluding ectopic ACTH syndrome in children with unvisible adenoma on pituitary magnetic resonance imaging (MRI).

Atrial fibrillation (AF), the most common cardiac arrhythmia is associated with increased morbidity and mortality. The higher mortality is due to the risk of heart failure and cardioembolic events. This in-depth review focuses on the strategies and efficacy of catheter ablation for non-paroxysmal atrial fibrillation. The main medical databases were searched for contemporary studies on catheter ablation for non-paroxysmal AF. Catheter ablation is currently proven to be the most effective treatment for AF and consists of pulmonary vein isolation as the cornerstone plus additional ablations. In terms of SR maintenance, it is less effective in non-paroxysmal AF than in paroxysmal patients. but the clinical benefit in non-paroxysmal patients is substantially higher. Since pulmonary vein isolation is ineffective, a variety of techniques have been developed, e.g. linear ablations, ablation of complex atrial fractionated electrograms, etc. Another paradox consists in the technique of catheter ablation. Despite promising results in early observation studies, further randomized studies have not confirmed the initial enthusiasm. Recently, a new approach, pulsed-field ablation, appears promising. This is an in-depth summary of current technologies and techniques for the ablation of non-paroxysmal AF. We discuss the benefits, risks and implications in the treatment of patients with non-paroxysmal AF.

Aims: To investigate DNA methylation of specific gene promoters in endometrial hyperplasia compared to normal endometrial tissue.

Materials and methods: To search for epigenetic events, methylation-specific multiplex ligation-dependent probe amplification was employed to compare the methylation status of 64 tissue samples with atypical endometrial hyperplasia, 60 tissue samples with endometrial hyperplasia without atypia, and 40 control tissue samples with normal endometrium.

Results: Differences in DNA methylation among the groups were found in PTEN, CDH13, and MSH6 promoters (PTEN: atypical hyperplasia 32%, benign hyperplasia 6.8%, normal endometrium 10%; P=0.004; CDH13: atypical hyperplasia, 50%; benign hyperplasia, 43%; normal endometrium 8.1%; P=0.003; MSH6 atypical hyperplasia 84%, benign hyperplasia, 62%; normal endometrium, 52%; P=0.008.) Higher rates of CDH13 promoter methylation were identified in the groups with both forms of endometrial hyperplasia when compared to the control group (atypical hyperplasia, P=0.003, benign hyperplasia, P=0.0002). A higher rate of DNA methylation of the PTEN and MSH6 promoters was observed in samples with atypical endometrial hyperplasia than in samples with benign endometrial hyperplasia (PTEN: P=0.02; MSH6: P=0.01) and samples with normal endometrial tissue (PTEN, P=0.04; MSH6, P=0.006).

Conclusion: DNA methylation of CDH13, PTEN, and MSH6 appear to be involved in the development of endometrial hyperplasia.

Aims: The objective of this study was to investigate the association and combined prognostic significance of the PD-L1, Smoothened protein and β-catenin expressions in patients with clear cell renal cell carcinoma (ccRCC).

Methods: The PD-L1, Smoothened protein and β-catenin expression were evaluated in 104 ccRCC patients. All studied tumor samples were acquired from nephrectomy specimens of primary tumors and not from biopsies or metastases. An indirect immunohistochemistry using polyclonal rabbit anti-Smoothened antibody, monoclonal mouse anti-human β-catenin-1 antibody, immunohistochemical assay PD-L1 28-8 pharmDx using monoclonal rabbit anti-PD-L1 antibody and anti-VHL (C- terminal) rabbit antibody was used. Immunohistochemistry was scored semiquantitavely.

Results: Median overall survival (OS) was significantly better in patients with lower PD-L1 expression (≤5%), Smoothened protein (SMO) expression (<5%) or cytoplasmic β-catenin expression (≤75%) than in patients with higher expressions of these biomarkers (P<0.001, P=0.047, and P<0.001, respectively). Membranous β-catenin showed an opposite effect with its lower expression (≤75%) being associated with longer OS (P=0.020). There was significant association between PD-1 and PD-L1 expression (P=0.007) and significant association of tumor grade (WHO 2016) with membranous β-catenin (P<0.001), cytoplasmic β-catenin (P=0.005), pVHL (P=0.042), PD-L1 (P=0.049) and PD-1 (P=0.028) expression.

Conclusion: The present study provides the first data on the potential association and combined prognostic significance of frequency of primary cilia, PD-L1, Smoothened protein and β-catenin expression with the outcome in clear cell renal cell carcinoma.

Aim: The purpose of this project was to compare the characteristics of two experimental murine models of primary intraocular lymphoma (PIOL) and determine which experimental model is most suitable for further investigational research to elucidate the pathophysiology of PIOL and to find new therapeutical strategies.

Methods: In both experimental models PIOL was induced in immunocompetent mice with intravitreal injection of syngeneic B-cell lymphoma cell lines. Murine strain C3H/HeN and cell line 38C13 were used in the first model and BALB/CaNn mice and cell line A20 in the second model. During the experiments, thorough clinical evaluation (using photo documentation, ultrasonography, and MRI) and histological evaluation were performed.

Results: In both models, the percentage of PIOL development was high, reaching nearly 80%. Disease progression was faster in C3H/HeN with exophthalmos occurring on average on day 10. Vitreous involvement was a predominant sign in the clinical presentation of this group. In BALB/CaNn mice exophthalmos occurred on average on day 22. The predominant clinical sign in the BALB/CaNn group was tumorous infiltration of the retina, optic disc, and tumorous retinal detachment.

Conclusion: Slower progression of the disease in BALB/CaNn mice, greater possibility to examine the retina due to mild vitreous involvement, and later occurrence of exophthalmos makes this strain more suitable for further investigational research.

Background: Oxidative stress and inflammation are considered predictors of diseases associated with aging. Markers of oxidative stress, inflammation, and endothelial activation were investigated in people with HIV on antiretroviral treatment to determine whether they had an immunosenescent phenotype that might predispose to the development of premature age-related diseases.

Patients and methods: This study was conducted on 213 subjects with HIV. The control groups consisted of healthy HIV-negative adults. The level of oxidative stress was measured by assessing the production of malondialdehyde levels, which were detected by thiobarbituric acid reactive substance (TBARS) assay. The level of microparticles indicated the presence of inflammation and endothelial activation was measured by E-selectin levels. Significant differences were determined by appropriate statistical tests, depending on the distribution of variables. Relationships between continuous variables were quantified using Spearman's rank correlation coefficient.

Results: TBARS, and microparticle and E-selectin levels were significantly higher in untreated and treated subjects with HIV compared with HIV-negative controls (P<0.001). The levels of the investigated markers were not significantly different between untreated and treated patients and no significant correlation of these markers was found with CD4⁺ count, CD4⁺/CD8⁺ ratio, and the number of HIV-1 RNA copies.

Conclusions: Elevated markers of oxidative stress, inflammatory and endothelial activation were independent of the virologic and immunologic status of people with HIV. These results support the hypothesis that residual viremia in cellular reservoirs of various tissues is a key factor related to the premature aging of the immune system and predisposition to the premature development of diseases associated with aging.

Aim: To assess nutritional status and muscle mass loss in patients with chronic obstructice pulmonary disease (COPD) from the Czech National Database of COPD and to evaluate the association of nutritional parameters with COPD phenotype, lung function, COPD-related symptoms and long-term survival.

Methods: A total of 343 patients with known body composition parameters - body mass Index (BMI), fat-free mass index (FFMI) and mid-art muscle circumference (MAMC) - were included in the analysis. Descriptive statistical methods were used to assess differences between groups, and overall survival was assessed using the Kaplan-Meier method after 9 years of follow-up.

Results: Nutritional imbalances were common in patients with COPD. Underweight and muscle mass loss were associated with emphysematous and pulmonary cachexia phenotypes, worse lung function, more symptoms and exacerbations, osteoporosis and depression. Overweight and obesity were associated with a chronic bronchitis phenotype and cardiovascular and metabolic comorbidities. Obese patients had higher forced exspiratory volume in 1 second (FEV₁) and diffusing capacity of the lung for carbon monoxid (DL_CO), but lower forced exspiratory capacity (FVC). Malnutrition increased the risk of death. Cachexia and underweight reduced median survival from 72.0 to 43.5 (P=0.049) and 39.8 (P=0.010) months, respectively, compared to normal nutrition. Muscle mass loss by MAMC of ≥20% was the strongest predictor of mortality, increasing the risk of death by 5.5-fold compared to patients with normal muscle mass (P<0.001). Patients with overweight and obesity had a 40% (P=0.026) and 30% lower risk of death, respectively, compared to patients with normal nutrition.

Conclusions: Patients with COPD often have nutritional imbalances. That is an important determinant of clinical characteristics and outcomes in patients with COPD. Further research is needed to better understand these differences.

Purpose: The aim of this study was to define the thickness of the retinal nerve fiber layer (RNFL) in the peripapillary region of the retina after adjusting for the effect of vessel density (VD) in patients with pathological intraocular pressure (IOP).

Patients and methods: 69 patients (122 eyes) with IOP >21 mmHg (range 21-36 mmHg, mean 23.65±2.70 mmHg). 32 were men (average age 55±13 years) and 37 were women (average age 52±14 years). IOP was measured using the Ocular Response Analyser (ORA). VD and RNFL were measured peripapillary by OCT (Avanti RTVue XR) in eight segments: Inferior Temporal - IT (1); Temporal Inferior - TI (2); Temporal Superior -TS (3); Superior Temporal - ST (4); Superior Nasal - SN (5); Nasal Superior - NS (6); Nasal Inferior - NI (7) and Inferior Nasal - IN (8). The VD value was subtracted from the total RNFL value.

Results: A corrected value for the RNFLc nerve fiber layer thickness (RNFLc) was introduced to account for VD across the RNFL volume in each segment. Person's correlation coefficient (r) was used to assess the correlation between IOP and RNFLc. The strongest correlations in RNFLc were in segments 5 (r=-0.32, P=0.002) and 8 (r=-0.21, P=0.037).

Conclusion: The greatest changes in RNFLc (RNFL minus VD) were in eyes with pathological IOP in segments 5 and 8, the location of the retinal ganglion cell magnocellular fibers. That is, when the thickness of the nerve fiber layer was reduced by correcting for vessel density, there was a significant correlation in segments 5 (r =-0.32, p<0.05) and 8 (r =-0.21, P<0.05) with intraocular pressure. The results suggest use of a corrected RNFL from VD value as more appropriate for detecting early changes in glaucoma.

Introduction: Macular pigment plays an important role in the reduction of oxidative stress and in preventing retinal diseases. Quick and easy measurements of the macular pigment are essential in both clinical and research settings. Dual wavelength fundus auto-fluorescence seems to be the optimal method. This study aims to investigate changes in fundus autofluorescence in patients taking daily lutein oral supplements and develop image processing methods for follow-up evaluations of the images.

Methods: New devices allow us to examine fundus autofluorescence using both blue and green excitation wavelengths. This allows detection of the amount of macular pigment by subtracting these two images because the yellow pigment particles absorb blue wavelengths. We determined daily dose of 25 mg of lutein and 3 mg of zeaxanthin. Patients were followed up for 15 months at 3-month intervals.

Results: During our 15-month study, we observed a positive trend in pixel lightness values, suggesting an increase in macular pigments in the foveal area. In all patients taking daily lutein supplements, the foveal index significantly increased after six months, with a median change of 0.081. We did not observe a significant change after the first three months (0.006) and only a small change between the 6th and 12th-month visits (0.012).

Conclusion: With appropriate patients and procedures for capturing autofluorescence images, this is a valuable technique for macular pigment evaluation in follow-up examinations using software image post-processing and analysis with commonly available hardware. To put this into everyday practice, developing tools to automate the assessment is necessary.