Biomedical Papers-Olomouc最新文献

Aims/background: Johanson-Blizzard syndrome (JBS) is a rare autosomal recessive disease caused by pathogenic variants in the UBR1 gene. JBS is usually suspected based on characteristic anomalies, but only genetic testing provides a definitive diagnosis. Since most variants are inherited from the parents, we aimed to identify the causal variants in a Czech proband with clinically suspected JBS and perform segregation analysis.

Methods: A proband with clinically suspected JBS underwent clinical exome sequencing (CES). Sanger sequencing was used for the validation, characterization, and segregation of variants in the family. The variants were also characterized using quantitative real-time PCR (qPCR) and in silico analysis.

Results: Using CES in the proband, we identified two novel causal variants in the UBR1 gene, c.3482A>C and c.3509+6T>C. Although the variants were found in trans, neither was detected in the parents. Sanger sequencing of the cDNA revealed that the novel variant c.3509+6T>C caused activation of the non-canonical GC donor splice site. The inclusion of 70 bp of the intronic sequence generated a frameshift and a premature termination codon leading to nonsense-mediated decay, as detected by qPCR. In silico protein structural analysis showed that the novel missense variant c.3482A>C in the zinc-stabilized domain RING-H2 altered a highly conserved zinc-coordinating histidine by proline.

Conclusion: To the best of our knowledge, we report the first molecular confirmation of JBS in the Czech Republic and the first identification of two de novo causal variants in two alleles. Our findings also expand the spectrum of pathogenic variants in the UBR1 gene.

Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) are crucial diagnostic modalities that require patients to remain immobile for extended periods, with anesthesia sometimes used for comfort and image quality enhancement. The study compares dexmedetomidine and propofol in reducing recovery time and sedation onset in pediatric and elderly patients undergoing CT and MRI procedures. A meta-analysis of fifteen studies assessing recovery time, sedation onset, and failed sedation between dexmedetomidine and propofol in pediatric and elderly patients during CT and MRI was conducted. The study indicated that the administration of anaesthesia markedly improved patient compliance and reduced motion artefacts in both CT and MRI (P<0.00001, I²=94%). The meta-analysis indicated that the mean difference (MD) in the onset of sedation was significantly faster in the control group (P<0.00001, I²=96%). The study reveals that dexmedetomidine and propofol anesthesia can improve patient image quality during CT and MRI procedures by reducing motion artefacts. Dexmedetomidine sedated people more quickly than propofol, but no significant differences in sedation duration were observed.

Background and aims: Neurodegenerative disorders affecting the brain and spinal cord are caused by a large number of factors. More recently, imbalances in gut microbiota are found to be one factor linked directly to neurological dysfunction. Probiotics prevent cognitive decline. For the first time, the effect of probiotics was assessed by monitoring the concentrations of the neurodegeneration biomarker neurofilament light chains (NfL) in a well-defined group of community-dwelling individuals. The aim of this study was to determine whether administration of our new probiotics could reduce NfL concentrations.

Methods: The serum NfL concentrations were measured in total of 190 serum samples of 85 older community-dwelling individuals. The participants were randomly divided into two groups: the PROPLA group and the PLAPRO group. Individuals in the PROPLA group started with a three-month use of probiotics and continued with a three-month use of placebo while the order was reversed in the PLAPRO group. The participants underwent detailed examinations at three time points: at baseline, in three and six months. The serum NfL concentrations were determined using ultrasensitive single-molecule array (SIMOA) assay.

Results: Longitudinal comparisons of NfL concentrations between samplings at different time points in the PROPLA and PLAPRO groups showed no statistically significant differences. Baseline NfL concentrations at the beginning of the study and in the succeeding samplings were not significantly different for the two groups in cross-sectional comparisons.

Conclusions: Serum NfL concentrations were not influenced by the three-month use of probiotics.

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.

Background: Left ventricular thrombus (LVT) formation is one of the well-known and serious complications of acute myocardial infarction (AMI) due to the risk of systemic arterial embolization (SE). To diagnose LVT, echocardiography (TTE) is used. Late gadolinium-enhanced cardiovascular magnetic resonance (DE-CMR) is the gold standard for diagnosing LVT.

Objectives: The aim of this observational study was to determine the role of transthoracic echocardiography and cardiac markers in predicting the occurrence of LVT compared with a reference cardiac imaging (DE-CMR) and to determine the risk of systemic embolization to the CNS using brain MRA.

Methods: Seventy patients after MI managed by percutaneous coronary intervention (localization: 92.9% anterior wall, 7% other; median age 58.7 years) were initially examined by transthoracic echocardiography (TTE, n=69) with a focus on LVT detection. Patients were then referred for DE-CMR (n=55). Laboratory determination of cardiac markers (Troponin T and NTproBNP) was carried out in all. Brain MRA was performed 1 year apart (n=51).

Results: The prevalence of LVT detected by echocardiography: (n=11/69, i.e. 15.9%); by DE-CMR: (n=9/55, i.e. 16.7%). Statistically significant parameters to predict the occurrence of LVT after AMI (cut off value): (a) detected by echocardiography: anamnestic data - delay (≥ 5 hours), echocardiographic parameters - left atrial volume index (LAVI≥ 32 mL/m²), LV EF Simpson biplane and estimated (≤ 42%), tissue Doppler determination of septal A wave velocity (≤ 7.5cm/s); (b) detected by DE-CMR: anamnestic data - delay (≥ 13 hours), DE-CMR parameters - left ventricular end-diastolic diameter (≥ 54mm). The value of cardiac markers (Troponin T and NTproBNP in ng/L) in LVT detected by echocardiography did not reach statistical significance. In LVT detected by DE-CMR, NTproBNP was statistically significantly increased at 1 month after AMI onset (no optimal cut-off value could be determined). There was no statistically significant association between the LVT detection (both modalities) and the occurrence of clinically manifest and silent cardioembolic events.

Conclusion: Our study confirmed a relatively high prevalence of LVT in the high-risk group of patients with anterior wall STEMI. Due to the low prevalence of thromboembolic complications, no significant association between the LVT detection and the occurrence of a cardioembolic event was demonstrated.

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.

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.

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.

Aims: Fibromyalgia (FM) is a chronic, widespread musculoskeletal disease that is usually accompanied by hyperalgesia, fatigue, sleep disturbance and depression. Although the etiopathogenesis of this syndrome is unclear, it is presumed to have an inflammatory basis. There are currently no laboratory markers available to diagnose the condition. The aim of this study was to investigate potential biochemical markers that would prove to be valid, simple, routinely used, quickly and cheaply studied and obtainable in blood count tests.

Methods: 46 patients diagnosed with FM according to ACR (American College of Rheumatology) 2010 diagnostic criteria and 35 patients as a healthy control group were included in the study. Prealbumin, complete blood count, sediment and C-reactive protein (CRP) values of FM and control group patients were examined. Additionally, Hospital Anxiety and Depression Scale, Jenkins Sleep Scale and the Fibromyalgia Impact Questionnaire (FIQ) were administered to patients diagnosed with FM.

Results: There was a significant difference between fibromyalgia patients and control groups in terms of prealbumin, platelet, CRP, CRP/prealbumin and lymphocyte parameters (P=0.048, P=0.046, P<0.001, P=0.003 and P<0.001, respectively). Only a weak positive correlation was found between CRP and FIQ (r_S =0.309, P=0.037, CI=0.012-0.556).

Conclusion: The serum levels of some tested markers, including platelets, CRP, CRP/prealbumin and lymphocytes, were significantly higher and prealbumin was lower in patients with FM than in the control group, suggesting they could be valid in fibromyalgia diagnosis. The CRP level may be informative about the severity of the FM and it may play a role in the inflammation possibly underlying the pathogenesis of FM.

Background and aims: Chronic spontaneous urticaria (CSU) is a skin condition causing red, raised, itchy and sometimes painful hives which lasts longer than six weeks. While the cause is unclear, the Urticaria Activity Score-7 (UAS-7), can help to determine the severity of the disease while the Systemic Immune Inflammation Index (SII) defined as neutrophils × platelets/lymphocytes, is a recent marker that shows the inflammatory and immune status and can be easily calculated from routine blood tests using neutrophil, lymphocyte, and platelet counts. We aimed to determine whether the SII correlates with UAS-7 in CSU patients.

Methods: We conducted this study on 245 patients with CSU. The UAS-7 score was obtained from all patients. Four groups were generated according to the UAS-7 score as ≤6, 7-15, 16-27, and 28-42. The correlation between the UAS-7 score and SII was evaluated.

Results: There was a statistically significant difference between the groups in terms of white blood cell count, neutrophil count, platelet count, Neutrophil-lymphocyte ratio (NLR), Platelet- lymphocyte ratio (PLR), and SII (P<0.05). A positive correlation was found between SII, NLR and PLR, and UAS-7 score. The strongest correlation was between SII and UAS-7, with P<0.001, r=0.642.

Conclusion: SII, which is easy to calculate, practical, and objective can be used for predicting the severity of the urticaria and for guiding the follow-up and treatment of these patients.