Epilepsy, affecting over 50 million people globally, presents a significant neurological challenge. Effective prevention of epileptic seizures relies on proper administration and monitoring of Anti-Seizure Medication (ASMs). Therapeutic Drug Monitoring (TDM) ensures optimal dosage adjustment, minimizing adverse effects and potential drug interactions. While traditional venous blood collection for TDM may be stressful, emerging alternative sampling methods, particularly Dried Blood Spot (DBS) or oral fluid offer less invasive way of sampling. This study aimed to develop and validate an analytical method for the determination of lamotrigine in such alternative samples. The sample, either DBS or oral fluid, was subjected to extraction, evaporation, and reconstitution in 15 % acetonitrile containing 0.1 % formic acid. A Kinetex C18 Polar column was used for liquid chromatographic separation and MS in ESI+ mode was used for detection and quantitation of lamotrigine using an isotopically labelled internal standard according to EMA guidelines. The calibration range of the developed method enables the determination of lamotrigine in the concentration range of 1-30 μg/mL in DBS and 0.5-20 μg/mL in oral fluid. Oral fluid and DBS samples from patients treated with lamotrigine analysed by the developed method were compared to plasma concentrations measured by the hospital's accredited laboratory. Preliminary results indicate a promising potential for these alternative matrices in clinical TDM applications. By offering a less invasive sampling approach, this method improves the accessibility and safety of pharmacotherapy for epilepsy patients. The results of this study lay the foundation for further clinical applications by implementing alternative matrix TDM, which may significantly advance personalized care in epilepsy management.

• Klíčová slova
• Alternative matrix, Antiepileptics, Dried blood spot, Lamotrigine, Quantification, Saliva, Validation,
• MeSH
• antikonvulziva * analýza   krev   MeSH
• chromatografie kapalinová metody   MeSH
• epilepsie farmakoterapie   MeSH
• kalibrace MeSH
• kapalinová chromatografie-hmotnostní spektrometrie MeSH
• lamotrigin * analýza   krev   MeSH
• lidé MeSH
• limita detekce MeSH
• monitorování léčiv * metody   MeSH
• reprodukovatelnost výsledků MeSH
• sliny * chemie   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• test suché kapky krve * metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• validační studie MeSH
• Názvy látek
• antikonvulziva * MeSH
• lamotrigin * MeSH

There is a lack of reliable in vivo models that replicate limb-threatening ischemia in humans. To fill this gap, we developed and validated two novel porcine ischemic models: ischemic limb and dorsal flap models, both with and without streptozotocin-induced hyperglycemia (N = 3 per group, 12 in total). Hind limb ischemia model was induced via different arterial ligations, with two ischemic and three control wounds per animal. In the flap model, four full-thickness flaps were created on the dorsum with silicone sheets to block reperfusion, and excisional wounds were made on the top. One non-ischemic wound served as control. Transcutaneous oxygen pressure (TcPO2), wound area, and microvascular density were measured, with TcPO2 and wound area assessed longitudinally. Data analysis focused on detailed visualization and Bayesian hierarchical modelling to account for the small sample size. Developed models exhibited stable ischemia and prolonged wound healing, with TcPO2 remaining under 30 mmHg over 28 days, and wound healing extending beyond two weeks. The flap model showed slower TcPO2 recovery and greater chronicity compared to the limb model, without reliable effect of hyperglycemia. Thus, the porcine flap model shows the highest potential as a relevant model for chronic limb-threatening ischemia.

• Klíčová slova
• Animal model, Bayesian modelling, Critical limb ischemia, Diabetes mellitus, In vivo model, Pig,
• MeSH
• časové faktory MeSH
• chirurgické laloky * MeSH
• chronická kritická ischemie končetin * patofyziologie   patologie   krev   MeSH
• chronická nemoc MeSH
• experimentální diabetes mellitus komplikace   krev   chemicky indukované   MeSH
• fyziologická neovaskularizace MeSH
• hojení ran MeSH
• hyperglykemie chemicky indukované   komplikace   krev   MeSH
• ischemie * patofyziologie   patologie   krev   etiologie   MeSH
• krevní glukóza metabolismus   MeSH
• mikrovaskulární denzita MeSH
• modely nemocí na zvířatech MeSH
• prasata MeSH
• regionální krevní průtok MeSH
• reprodukovatelnost výsledků MeSH
• Sus scrofa MeSH
• transkutánní měření krevních plynů MeSH
• zadní končetina krevní zásobení   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• validační studie MeSH
• Názvy látek
• krevní glukóza MeSH

Monitoring patients with phenylketonuria (PKU) requires accurately measuring phenylalanine and tyrosine levels in a small volume of blood samples obtained by the dried blood spot (DBS) technique. The aim was to study selected parameters influencing the quantitative results. Phenylalanine and tyrosine were extracted from DBS samples with methanol, and 5 internal standard introduction techniques were tested. Phenylalanine and tyrosine levels were measured in 6-mm discs punched from DBS, pre-punched 9-mm discs containing the entire DBS sample, and liquid blood by HPLC-MS-MS. Levels in 6-mm discs punched from DBS measured by HPLC-MS/MS were compared with those measured by the HPLC-FLD. The analytical parameters of the method are satisfactory, linearity in the range of 25-1200 μmol/L (LOD, LOQ and LLOQ values 0.2 μmol/L, 0.5 μmol/L and 3.8 μmol/L for phenylalanine, 0.5 μmol/L, 1.5 μmol/L and 5.1 μmol/L for tyrosine), within-run precision 1.8 %-3.7 % for phenylalanine, 1.9 %-2.7 % for tyrosine, between-run precision 4.7 %-5.9 % for phenylalanine, 4.1 %-5.4 % for tyrosine, recovery 93.8 %-100.4 % for phenylalanine and 93.7 %-99.1 % for tyrosine. Good agreement was found between phenylalanine and tyrosine concentrations in 6-mm discs punched from DBS (R = 0.896, p < 0.001, and R = 0.907, p < 0.001, respectively), pre-punched 9-mm discs containing the entire DBS sample (R = 0.960, p < 0.001, and R = 0.950, p < 0.001, respectively) and liquid blood, as well as between phenylalanine and tyrosine concentrations obtained by HPLC-MS/MS and HPLC-FLD (R = 0.968, p < 0.001, and R = 0.984, p < 0.001, respectively). The presented method is suitable for monitoring patients with PKU.

• Klíčová slova
• Dried blood spot, Liquid chromatography with mass spectrometry and fluorescence detection, Parameters affecting quantitative results, Phenylalanine, Tyrosine,
• MeSH
• fenylalanin * krev   MeSH
• fenylketonurie * krev   diagnóza   MeSH
• lidé MeSH
• limita detekce MeSH
• reprodukovatelnost výsledků MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• test suché kapky krve * metody   MeSH
• tyrosin * krev   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fenylalanin * MeSH
• tyrosin * MeSH

As a chronic metabolic disease, diabetes mellitus necessitates ongoing self-management to control blood sugar levels and avoid complications. Although diabetes management relies heavily on technologies like insulin pumps and continuous glucose monitors (CGMs), patient use of these devices may differ depending on personality factors. This study investigated the association between diabetes types, personality types, and preferences for diabetes management technology. The 56 patients receiving intense insulin therapy completed the Myers-Briggs Type Indicator (MBTI) to evaluate personality qualities. Multinomial logistic regression was employed for statistical analysis after technology utilization data was gathered. According to the findings, introverts are more likely to utilize CGMs. Insulin pumps were preferred by extroverts, indicating their inclination for proactive, hands-on care. There were no significant relationships between diabetes type, technology use, or other aspects of personality. These results imply that personality variables affect technology preferences and could be helpful in customizing diabetes care plans. This study highlights the importance of considering psychological issues when choosing diabetes solutions.

• Klíčová slova
• Continuous glucose monitoring, Diabetes management technologies, Insulin pumps, MBTI, Personality types, Technology preferences,
• MeSH
• diabetes mellitus * psychologie   terapie   MeSH
• dospělí MeSH
• inzulinové infuzní systémy * statistika a číselné údaje   MeSH
• lidé středního věku MeSH
• lidé MeSH
• osobnost * MeSH
• péče o sebe * MeSH
• selfmonitoring glykemie * statistika a číselné údaje   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Autologous cell therapy (ACT) is a new therapeutic approach for diabetic patients with no-option chronic limb-threatening ischemia (NO-CLTI). The aim of our study was to quantify cell populations of cell therapy products (CTPs) obtained by three different isolation methods and to correlate their numbers with changes in transcutaneous oxygen pressure (TcPO2). CTPs were separated either from stimulated peripheral blood (PB) (n = 11) or harvested from bone marrow (BM) processed either by Harvest SmartPReP2 (n = 50) or sedimented with succinate gelatin (n = 29). The clinical effect was evaluated by the change in TcPO2 after 1, 3 and 6 months. TcPO2 increased significantly in all three methods at each time point in comparison with baseline values (p < .01) with no significant difference among them. There was no correlation between the change in TcPO2 and the size of injected cell populations. We only observed a weak correlation between the number of injected white blood cells (WBC) and an increase in TcPO2 at 1 and 3 months. Our study showed that all three isolation methods of ACT were similarly relatively efficient in the treatment of NO-CLTI. We observed no correlation of TcPO2 increase with the number of injected monocytes, lymphocytes or CD34+. We observed a weak correlation between TcPO2 increase and the number of injected WBCs.

• Klíčová slova
• autologous cell therapy, cell therapy product, chronic limb-threatening ischemia, transcutaneous oxygen pressure,
• MeSH
• autologní transplantace metody   MeSH
• buněčná a tkáňová terapie * metody   MeSH
• chronická kritická ischemie končetin * terapie   diagnóza   chirurgie   MeSH
• ischemie * terapie   etiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• transkutánní měření krevních plynů metody   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Objective: The recommended threshold for the time spent on continuous glucose monitoring (CGM) is established at 70%. However, glucose outcomes in children with type 1 diabetes (CwD) using CGM for a different proportion of time within this threshold have not been evaluated yet. The study aims to compare glycemic parameters among CwD who spent 70%-89% and ≥90% on CGM using the population-wide data from the Czech national pediatric diabetes registry ČENDA. Methods: CwD aged <19 years who used real-time CGM >70% of the time and did not change the type of therapy throughout the year 2023 were included and divided into two groups based on the time they spent on CGM-70%-89% versus ≥90%. HbA1c, times in standard glycemic ranges, mean glucose, and coefficient of variability (CV) were compared between the groups and by treatment modalities. Results: Data from 1977 CwD (1035 males and 942 females) were evaluated. Among them, 404 participants (20.4%) used CGM 70%-89% of the time, and 1573 participants (79.6%) ≥90% of the time. Compared with the 70-89% group, the ≥90% CGM users achieved significantly lower HbA1c levels (51 mmol/mol, 6.8% vs. 58 mmol/mol, 7.4%, P < 0.001), higher time in range (72% vs. 60%, P < 0.001), and lower mean glucose and CV (8.1 mmol/L, 146 mg/dL vs. 9.1 mmol/L, 164 mg/dL and 37% vs. 40%, respectively, both P < 0.001). Analogous results were seen irrespective of the treatment modality. The differences persisted after propensity score adjustment. Conclusion: CGM use for ≥90% is associated with tighter glycemic control compared with 70%-89% use. Therefore, it is essential to motivate CwD to use CGM for the longest possible time and search for suitable options to overcome barriers in uninterrupted CGM monitoring.

• Klíčová slova
• CGM, glucose monitoring, pediatrics, registry, type 1 diabetes,
• MeSH
• časové faktory MeSH
• diabetes mellitus 1. typu * krev   farmakoterapie   MeSH
• dítě MeSH
• glykovaný hemoglobin analýza   MeSH
• hypoglykemika terapeutické užití   MeSH
• kontinuální monitorování glukózy MeSH
• krevní glukóza * analýza   MeSH
• lidé MeSH
• mladiství MeSH
• předškolní dítě MeSH
• registrace MeSH
• regulace glykemie * metody   MeSH
• selfmonitoring glykemie * metody   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• glykovaný hemoglobin MeSH
• hemoglobin A1c protein, human MeSH Prohlížeč
• hypoglykemika MeSH
• krevní glukóza * MeSH

INTRODUCTION: There is a broad awareness of shifts in the oxygen hemoglobin dissociation (ODC) relationship associated with fetal hemoglobin (HbF) changes. However, quantification of the shift has been limited. Aim was to quantify the shift of partial oxygen tension (PO2) associated with HbF and with changes after transfusion of adult erythrocytes (TAE) in preterm infants. METHODS: This is a single-center, retrospective observational analysis of blood gas samples. The shifts of ODC and PO2 related to HbF were evaluated in two models. Either HbF or TAE status (0, 1, ≥2) were used as the independent variable. Multivariate analysis was used to correct for confounding effects (gestational age, postnatal age, source of blood gas sample as well as pH, SO2, and PCO2). RESULTS: There were 3,452 blood gas observations analyzed from 2,464 infants whose median gestational age was 334 weeksdays (IQR 296-363). With SpO2 between 90 and 95%, the ODC was shifted to the left (13 mm Hg, 1.3 kPa). After adjusting for confounding variables, the number of TAEs (0, 1, ≥2), was highly significantly related to a shift (p < 0.001), consistent with the percent HbF level (p < 0.001). Based on the multivariate model (i.e., holding confounding parameters constant), with a SpO2 of 92% the PaO2 could be expected to shift markedly higher with 2 or more TAEs in an extremely preterm infant (7.3 mm Hg, 0.97 kPa). CONCLUSION: While preliminary, these data suggest that in vulnerable preterm infants a change to a slightly lower SpO2 target range following TAE could maintain equivalent PaO2 exposure.

• Klíčová slova
• Erythrocyte transfusion, Fetal hemoglobin, Neonatal oxygen targeting,
• MeSH
• analýza krevních plynů MeSH
• fetální hemoglobin * metabolismus   analýza   MeSH
• gestační stáří MeSH
• kyslík * krev   MeSH
• lidé MeSH
• novorozenec nedonošený * krev   MeSH
• novorozenec MeSH
• retrospektivní studie MeSH
• saturace kyslíkem * MeSH
• transfuze erytrocytů * škodlivé účinky   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• Názvy látek
• fetální hemoglobin * MeSH
• kyslík * MeSH

The study focused on the changes in C-peptide, glycemia, insulin concentration, and insulin resistance according to LDL-cholesterol concentration ranges. The metabolic profile of individuals in the Czech Republic (n = 1840) was classified by quartiles of LDL-cholesterol into four groups with the following ranges: 0.46-2.45 (n = 445), 2.46-3.00 (n = 474), 3.01-3.59 (n = 459), and 3.60-7.18 mmol/l (n = 462). The level of glucose, C-peptide, insulin, and area of parameters during OGTT and HOMA IR were compared with a relevant LDL-cholesterol range. The evaluation involved correlations between LDL-cholesterol and the above parameters, F-test and t-test. Generally, mean values of glucose homeostasis-related parameters were higher with increasing LDL-cholesterol levels, except for mean HOMA IR values which rapidly increased (2.7-3.4) between LDL-cholesterol ranges of 3.00-3.59 and 3.60-7.18 mmol/l. Glucose, C-peptide, insulin concentrations, and the area of parameters reached greater changes especially after glucose load during OGTT (p ≤ 0.001). Considerable changes were already observed for the above parameters between groups with LDL-cholesterol ranges of 2.46-3.00 and 3.01-3.59 mmol/l. HOMA IR increased with higher LDL-cholesterol concentrations, but the differences in mean values were not statistically significant. Most important differences appeared in glucose metabolism at LDL-cholesterol concentrations of 3.60-7.18 mmol/l in comparison to LDL-cholesterol lower ranges. In particular, the areas of C-peptide, glucose, and insulin ranges showed statistically significant differences between all groups with growing LDL-cholesterol ranges. The variances of HOMA IR statistically differed between groups created according to LDL-cholesterol concentrations ranges.

• Klíčová slova
• C-peptide, Glucose, HOMA IR, Insulin, LDL-cholesterol,
• MeSH
• C-peptid krev   metabolismus   MeSH
• diabetes mellitus * krev   diagnóza   metabolismus   prevence a kontrola   MeSH
• dospělí MeSH
• glukózový toleranční test MeSH
• hyperlipidemie * krev   diagnóza   metabolismus   prevence a kontrola   MeSH
• inzulin krev   metabolismus   MeSH
• inzulinová rezistence MeSH
• krevní glukóza * analýza   metabolismus   MeSH
• LDL-cholesterol * krev   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• C-peptid MeSH
• inzulin MeSH
• krevní glukóza * MeSH
• LDL-cholesterol * MeSH

AIM: To determine whether people with type 1 diabetes (T1D) initiating glucose sensor monitoring experience greater improvements in HbA1c when provided with education on carbohydrate counting and flexible insulin dosing than those who do not receive nutrition education. MATERIALS AND METHODS: Our retrospective observational study included 329 people with T1D initiating glucose sensor monitoring between 2015 and 2021. The participants were divided into two groups: one group attended at least one structured educational session with a registered dietitian (n = 126), while the other group did not receive structured education (n = 203). After 12 months of glucose sensor initiation, we compared glycaemic outcomes and CGM metrics between the two groups. RESULTS: At glucose sensor initiation, both groups with and without education had similar HbA1c levels (7.64% [60.0 mmol/mol] vs. 7.66% [60.2 mmol/mol]). After twelve months, the education group demonstrated greater improvement in glycemic outcomes (HbA1c 7.17% [54.9mmol/mol] vs. 7.37% [57.1 mmol/mol], p < 0.05) and spent significantly more time in the target range than did the group without structured education (68.8% vs. 64.1%, p < 0.05). We observed an inverse correlation between the number of completed educational sessions and HbA1c after 12 months, as well as between the number of educational sessions and the change in HbA1c. CONCLUSIONS: People with T1D who initiated glucose sensor monitoring alongside nutrition education showed greater improvements in HbA1c and increased time spent in the target glucose range compared to individuals who did not receive structured education. TRAIL REGISTRATION: ClinicalTrials.gov identifier: NCT06264271.

• Klíčová slova
• glucose sensors, glycemic outcomes, nutrition education, type 1 diabetes,
• MeSH
• diabetes mellitus 1. typu * krev   farmakoterapie   dietoterapie   MeSH
• dospělí MeSH
• glykovaný hemoglobin analýza   MeSH
• hypoglykemika aplikace a dávkování   terapeutické užití   MeSH
• inzulin aplikace a dávkování   MeSH
• krevní glukóza * analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• regulace glykemie * metody   MeSH
• retrospektivní studie MeSH
• selfmonitoring glykemie * metody   MeSH
• vzdělávání pacientů jako téma * metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• Názvy látek
• glykovaný hemoglobin MeSH
• hemoglobin A1c protein, human MeSH Prohlížeč
• hypoglykemika MeSH
• inzulin MeSH
• krevní glukóza * MeSH

BACKGROUND: Large vessel carotid stenosis is a significant cause of ischaemic stroke. Indications for surgical revascularisation depend on the severity of the stenosis and clinical symptoms. However, mild symptoms such as TIA (Transient ischaemic attack), amaurosis fugax or minor stroke precede large strokes in only 15% of cases. AIM: The aim of this prospective study is to evaluate whether retinal perfusion is impacted in significant carotid stenosis. Automated retinal oximetry will be used to better assess perfusion in the post-stenotic basin. We presume the more stenotic the blood vessel, the more reduced the retinal perfusion is, resulting in adaptive changes such as greater arteriovenous saturation difference due to greater oxygen extraction. This could broaden the indication spectrum for revascularisation for carotid stenosis. METHODS: We plan to enroll yearly 50 patients with significant carotid stenosis and cross-examine them with retinal oximetry. The study group will provide stenotic vessels and, non-stenotic vessels will form the control group. Patients with significant carotid stenosis will undergo an MRI (Magnetic Resonnance imaging) examination to determine the presence of asymptomatic recent ischaemic lesions in the stenotic basin, and the correlation to oximetry parameters. STATISTICS: The stenosis severity and retinal oximetry parameters will be compared for study and control groups with a threshold of 70%, respectively 80% and 90% stenosis. Results will be then reevaluated with emphasis on MRI findings in the carotid basin. CONCLUSION: This prospective case control study protocol will be used to launch a multicentre trial assessing the relationship between significant carotid stenosis and retinal perfusion measured with automated retinal oximetry. Despite these differences, the findings indicate the potential of retinal oximetry for noninvasive real-time measurements of oxyhaemoglobin saturation in central nervous system vessels. Following calibration upgrade and technological improvement, verification retinal oximetry may potentially be applied to critically ill and anaesthesia care patients. The study on combined scanning laser ophthalmoscope and retinal oximetry supports the feasibility of the technique for oximetry analysis in newly born babies. TRIAL REGISTRATION: ClinicalTrials.gov, ID: NCT06085612.

• Klíčová slova
• arterio-venous difference, automated retinal oximetry, carotid stenosis, ischaemic stroke, magnetic resonance imaging,
• MeSH
• lidé MeSH
• oxymetrie * metody   MeSH
• prospektivní studie MeSH
• retina * patofyziologie   MeSH
• retinální cévy * patofyziologie   MeSH
• senioři MeSH
• stenóza arteria carotis * patofyziologie   diagnostické zobrazování   komplikace   MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• protokol klinické studie MeSH