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Oxidative stress is believed to play an important role in the pathogenesis of inflammatory bowel disease (IBD), specifically Crohn's disease (CD) and ulcerative colitis (UC). This meta-analysis aimed to identify and quantify the oxidative stress-related biomarkers in IBD and their associations with disease activity. We systematically searched Ovid MEDLINE, Ovid Embase, and Web of Science databases, identifying 54 studies for inclusion. Comparisons included: (i) active IBD versus healthy controls; (ii) inactive IBD versus healthy controls; (iii) active CD versus inactive CD; and (iv) active UC versus inactive UC. Our analysis revealed a significant accumulation of biomarkers of oxidative damage to biomacromolecules, coupled with reductions in various antioxidants, in both patients with active and inactive IBD compared to healthy controls. Additionally, we identified biomarkers that differentiate between active and inactive CD, including malondialdehyde, Paraoxonase 1, catalase, albumin, transferrin, and total antioxidant capacity. Similarly, levels of Paraoxonase 1, erythrocyte glutathione peroxidase, catalase, albumin, transferrin, and free thiols differed between active and inactive UC. Vitamins and carotenoids also emerged as potential disease activity biomarkers for CD and UC, but their intake should be monitored to obtain meaningful results. These findings emphasize the involvement of oxidative stress in the pathogenesis of IBD and highlight the potential of oxidative stress-related biomarkers as a minimally invasive and additional tool for monitoring the activity of IBD.

Abstract Background The aim of this double-blind, placebo-controlled study was to investigate the effect of vitamin E supplementation as an addition to a commercial renal diet on survival time of cats with different stages of chronic kidney disease (CKD). In addition, we were interested whether vitamin E supplementation affects selected oxidative stress and clinical parameters. Thirty-four cats with CKD and 38 healthy cats were included in the study. Cats with CKD were classified according to the IRIS Guidelines; seven in IRIS stage 1, 15 in IRIS stage 2, five in IRIS stage 3 and seven in IRIS stage 4. Cats with CKD were treated according to IRIS Guidelines. Cats with CKD were randomly assigned to receive vitamin E (100 IU/cat/day) or placebo (mineral oil) for 24 weeks in addition to standard therapy. Plasma malondialdehyde (MDA) and protein carbonyl (PC) concentrations, DNA damage of peripheral lymphocytes and plasma vitamin E concentrations were measured at baseline and four, eight, 16 and 24 weeks thereafter. Routine laboratory analyses and assessment of clinical signs were performed at each visit. Results Vitamin E supplementation had no effect on the survival time and did not reduce the severity of clinical signs. Before vitamin E supplementation, no significant differences in vitamin E, MDA and PC concentrations were found between healthy and CKD cats. However, plasma MDA concentration was statistically significantly higher (p = 0.043) in cats with early CKD (IRIS stages 1 and 2) than in cats with advanced CKD (IRIS stages 3 and 4). Additionally, DNA damage was statistically significantly higher in healthy cats (p ≤ 0.001) than in CKD cats. Plasma vitamin E concentrations increased statistically significantly in the vitamin E group compared to the placebo group four (p = 0.013) and eight (p = 0.017) weeks after the start of vitamin E supplementation. During the study and after 24 weeks of vitamin E supplementation, plasma MDA and PC concentrations and DNA damage remained similar to pre-supplementation levels in both the placebo and vitamin E groups. Conclusions Vitamin E supplementation as an addition to standard therapy does not prolong survival in feline CKD.

Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as mediators in cell–cell communication; however, their harvesting and assessment from bodily fluids are challenging, as processing can significantly affect samples. With the aim of minimizing processing artifacts, we assessed the number density (n) and hydrodynamic diameter (Dh) of EPs directly in diluted plasma and blood using the following recently developed technique: interferometric light microscopy (ILM). We analyzed 613 blood and plasma samples from human patients with inflammatory bowel disease (IBD), collected in trisodium citrate and ethylenediaminetetraacetic acid (EDTA) anticoagulants, and 163 blood and plasma samples from canine patients with brachycephalic obstructive airway syndrome (BOAS). We found a highly statistically significant correlation between n in the plasma and n in the blood only in the human (i.e., but not canine) blood samples, between the samples with trisodium citrate and EDTA, and between the respective Dh for both species (all p < 10−3). In the human plasma, the average was 139 ± 31 nm; in the human blood, was 158 ± 11 nm; in the canine plasma, was 155 ± 32 nm; and in the canine blood, was 171 ± 33 nm. The differences within species were statistically significant (p < 10−2), with sufficient statistical power (P > 0.8). For , we found no statistically significant differences between the human plasma and blood samples or between the samples with trisodium citrate and EDTA. Our results prove that measuring n and Dh of EPs in minimally processed fresh blood and in diluted fresh plasma by means of ILM is feasible for large populations of samples.

Background and Objectives: Oxidative stress resulting from a disturbance of the endogenous redox system is suspected in numerous diseases of the central nervous system, including epilepsy. In addition, antiseizure medications (ASMs), especially those of the old generation, may further increase oxidative stress. To evaluate the effects of ASM generation on oxidative stress, we conducted a cross-sectional study in patients with epilepsy treated with old, new, and polytherapy. Materials and Methods: The antioxidant activity of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, as well as the concentrations of malondialdehyde, protein carbonyl, nitrate, nitrite, and glutathione in reduced and oxidized forms, were measured in 49 patients with epilepsy and 14 healthy controls. In addition, the plasma concentrations of ASMs and metabolites of carbamazepine and valproic acid were measured in the patients. Results: Patients with epilepsy showed increased activities of superoxide dismutase and catalase (p < 0.001), concentrations of glutathione disulfide and markers of nitric oxide metabolism (p < 0.001), and decreased activities of glutathione peroxidase, glutathione reductase, glutathione, and nitrite concentrations (p ≤ 0.005) compared to healthy controls. A comparison of ASM generations revealed increased levels of superoxide dismutase and catalase (p ≤ 0.007) and decreased levels of glutathione peroxidase and glutathione reductase (p ≤ 0.01) in patients treated with old ASMs compared to those treated with new generation ASMs. In addition, an increase in protein carbonyl and nitric oxide metabolites (p ≤ 0.002) was observed in patients treated with old generation ASMs compared to those treated with new generation ASMs. Most oxidative stress parameters in patients receiving polytherapy with ASMs were intermediate between the results of patients treated with the old and new generations of ASMs. Conclusions: An increase in oxidative stress markers and modulation of antioxidant enzyme activities was observed in patients with epilepsy compared to controls. The results of our study showed significantly higher oxidative stress in patients treated with old ASMs compared to those treated with new generation ASMs.

Myxomatous mitral valve degeneration (MMVD) is the most common naturally occurring heart disease in dogs. There is a lack of data on antioxidant status and oxidative damage in dogs with MMVD stage B1 according to the American College of Veterinary Internal Medicine (ACVIM B1). The aim of this study was to investigate antioxidant status (plasma vitamin E, lipid-standardized vitamin E (LS-VitE), antioxidant capacity of lipid-(ACL) and water-soluble antioxidants, whole blood glutathione peroxidase and erythrocyte superoxide dismutase), and lipid peroxidation [malondialdehyde (MDA)] in dogs with MMVD ACVIM B1. Serum cholesterol and triglyceride concentrations were measured to calculate LS-VitE. Fourteen dogs with MMVD ACVIM B1 and 12 control dogs were included in the study. Dogs with MMVD had significantly higher vitamin E, ACL, MDA, and cholesterol concentrations and significantly higher LS-VitE values than control dogs. No significant correlations between MDA and antioxidant parameters were determined in either group. In conclusion, oxidative damage to lipids is already present and the antioxidant status is altered but not depleted in dogs with MMVD ACVIM B1. The antioxidant response to increased oxidative damage consists mainly of the activation of fat-soluble antioxidants. Further research is needed to evaluate the efficacy and targets of early antioxidant supplementation to prevent or ameliorate oxidative stress and mitigate disease progression in dogs with early-stage MMVD.

Background: The antiepileptic drug lamotrigine (LTG) shows high pharmacokinetic variability due to genotype influence and concomitant use of glucuronidation inducers and inhibitors, both of which may be frequently taken by elderly patients. Our goal was to develop a reliable quantification method for lamotrigine and its main glucuronide metabolite lamotrigine-N2-glucuronide (LTG-N2-GLU) in dried blood spots (DBS) to enable routine therapeutic drug monitoring and to identify altered metabolic activity for early detection of drug interactions possibly leading to suboptimal drug response. Results: The analytical method was validated in terms of selectivity, accuracy, precision, matrix effects, haematocrit, blood spot volume influence, and stability. It was applied to a clinical study, and the DBS results were compared to the concentrations determined in plasma samples. A good correlation was established for both analytes in DBS and plasma samples, taking into account the haematocrit and blood cell-to-plasma partition coefficients. It was demonstrated that the method is suitable for the determination of the metabolite-to-parent ratio to reveal the metabolic status of individual patients. Conclusions: The clinical validation performed confirmed that the DBS technique is a reliable alternative for plasma lamotrigine and its glucuronide determination.

The aim of the study was to investigate the effect of a therapeutic renal diet on selected clinical, biochemical, and urinary parameters and on selected parameters of oxidative stress in cats with early stages of chronic kidney disease (CKD). A prospective study of a 3-month duration was conducted to evaluate the effect of renal diet on selected clinical and laboratory parameters in client-owned cats with early stages of CKD. Of a total of 29 enrolled client-owned cats, nineteen (19) cats completed the study, ten receiving renal diet and nine receiving a diet of the owner’s choice. A clinical examination was performed, and blood and urine samples were collected on the day of presentation and at regular check-ups after 3-4, 7-8, and 10-12 weeks. Serum creatinine and symmetric dimethylarginine (SDMA) concentrations and selected parameters of oxidative stress (plasma glutathione peroxidase (GPX) activity and plasma malondialdehyde (MDA) and serum selenium concentrations), were measured and electrophoresis of urinary proteins was performed. At inclusion, a significant positive correlation (p < 0.001) was found between serum selenium concentration and plasma GPX activity (Pearson correlation coefficient 0.83 (95% CI: [0.65 - 0.92]) and a significant negative correlation (p < 0.001) between serum SDMA and urine specific gravity (Pearson correlation coefficient -0.70 (95% CI: [-0.87 - (-0.38)]). At the end of the 3-month feeding trial no significant difference was found in SDMA and creatinine concentrations. UČINEK MONOTERAPEVTSKE LEDVIČNE DIETE NA KLINIČNE, BIOKEMIJSKE, URINSKE IN SERUMSKE PARAMETRE OKSIDATIVNEGA STRESA PRI MAČKAH S CDK STOPNJE 1 IN 2 Izvleček: Namen študije je bil raziskati učinek terapevtske ledvične diete na izbrane klinične, biokemijske in urinske parametre ter parametre oksidativnega stresa pri mačkah v začetnih stopnjah kronične ledvične bolezni (KLB). Raziskava je bila zasnovana kot prospektivna, tri mesece trajajoča klinična študija, v katero je bilo vključenih 29 lastniških mačk. Devetnajst mačk je zaključilo študijo, od teh jih je deset prejemalo ledvično dieto, devet pa vzdrževalno dieto po izboru lastnika. Pri vseh mačkah smo izvedli klinični pregled in odvzem krvnih ter urinskih vzorcev na dan vključitve v študijo in pri treh kontrolnih pregledih, ki so bili izvedeni 3–4, 7–8 in 10–12 tednov kasneje. Določili smo serumsko koncentracijo kreatinina, simetričnega dimetilarginina (SDMA) in izbrane parametre oksidativnega stresa (aktivnost plazemske glutation peroksidaze (GPX) in plazemska koncentracija malondialdehida (MDA) ter serumska koncentracija selena). Poleg tega smo izvedli elektroforezo urinskih proteinov. Ob vključitvi mačk v raziskavo smo ugotovili značilno pozitivno korelacijo (p < 0,001) med serumsko koncentracijo selena in aktivnostjo plazemske GPX (Pearsonov korelacijski koeficient 0,83, 95 % CI: [0,65–0,92]) ter značilno negativno korealcijo (p < 0,001) med koncentracijo SDMA in specifično težo urina (Pearsonov korelacijski koeficient –0,70 (95 % CI: [–0,87–(–0,38)]). Trimesečno hranjenje s terapevtsko hrano ni privedlo do značilnih sprememb v serumski koncentraciji SDMA in kreatinina pri vključenih mačkah. Ključne besede: klinični parametri; simetrični dimetilarginin; oksidativni stres; ledvična dieta; mačke; kronična ledvična bolezen; elektroforeza urinskih proteinov

Clozapine is one of the most widely used second-generation antipsychotic drugs (SGAs) for the treatment of schizophrenia. Despite advantages over first-generation drugs, clozapine still shows significant side effects and interindividual variations in efficacy. In order to ensure frequent therapeutic drug monitoring (TDM) and improve the compliance of psychiatric patients undergoing clozapine treatment, two novel dried microsampling approaches based on whole blood and plasma volumetric absorptive microsampling (b-VAMS and p-VAMS) and microfluidic generated-dried blood spot technology (mfDBS) were developed and coupled to HPLC with electrochemical detection (ED). The proposed miniaturized strategies by means of VAMS and microfluidic channel-based devices provide several advantages in terms of collection, storage, and handling compared to classical blood and plasma processing. Satisfactory validation results were obtained for all microsampling platforms, with mean extraction yields >85.1%, precision as relative standard deviation (RSD) < 5.1%, and stability < 4.5% analyte loss after 30 days for p-VAMS; mean extraction yields > 83.4%, precision RSD < 5.4%, and stability < 4.6% analyte loss after 30 days for b-VAMS, and mean extraction yields > 74.0%, precision RSD < 5.6%, and stability < 4.9% analyte loss after 30 days for mfDBS. The original microsampling methodologies have been successfully applied to the blood and plasma collected from five psychiatric patients for the monitoring of the levels of clozapine and its main metabolites, providing robust and reliable quali-quantitative results. Comparisons between results of the two dried microsampling technologies with those obtained by classic fluid plasma analysis were in good agreement and have demonstrated that the proposed miniaturized approaches could be suitable for TDM purposes.

Abstract Background Antioxidants located in both the hydrophilic and lipophilic compartments of plasma act as a defence system against reactive oxygen species (ROS). Excessive production of ROS during anaesthesia affects the antioxidant capacity of plasma and may result in oxidative stress. The aim of this study was to evaluate the antioxidant capacity of lipid- (ACL) and water-soluble (ACW) antioxidants in client-owned dogs diagnosed with periodontal disease and early-stage myxomatous mitral valve degeneration (MMVD) and anaesthetised for a dental procedure with propofol and sevoflurane or with propofol only. Results Dogs with MMVD were anaesthetised with propofol and sevoflurane (MMVD/PS, n = 8) or with propofol only (MMVD/P, n = 10). Dogs with no evidence of MMVD (PS, n = 12) were anaesthetised with propofol and sevoflurane. Blood samples for determination of ACL and ACW were collected before and 5 min, 60 min and 6 h after induction to anaesthesia. In MMVD/PS dogs, ACL was significantly higher at all sampling times when compared to PS dogs. Compared to basal values, only anaesthesia maintained with propofol significantly increased ACL at 60 min in dogs with MMVD. In MMVD/P dogs, ACW increased after induction to anaesthesia and remained elevated up to 6 h after anaesthesia. Compared to basal values, anaesthesia maintained with sevoflurane significantly increased ACW only at 60 min in both dogs with and without MMVD. The only difference between propofol and propofol/sevoflurane anaesthesia in dogs with MMVD was significantly higher ACW at 60 min after induction to anaesthesia in the propofol group. Conclusions Regarding antioxidant capacity, propofol could be a better choice than sevoflurane for anaesthesia of dogs with early-stage MMVD, although further studies are necessary to clarify the advantage of this antioxidant capacity.

Canicross is a sport discipline that connects human and canine athletes in running. Changes in physiological, hematological, and biochemical parameters, and exercise-induced oxidative stress have not been thoroughly characterized in canicross dogs. The aim of our study was the assessment of the health status of trained canicross dogs that were subjected to two acute bouts of exercise with their owners during the training season. Health status was assessed by measuring the rectal temperature, hematological and biochemical parameters, as well as blood oxidative stress parameters (plasma malondialdehyde, lipid peroxidation marker; whole blood glutathione peroxidase and erythrocyte superoxide dismutase1, antioxidant enzymes) before and during a two-day canicross training session and after a 24-h rest period. Seven trained canicross dogs (three females/four males) aged 12–120 months were included in the study. Blood samples were collected before and immediately after the first acute bout of exercise (day 1), after the second acute bout of exercise (day 2), and after 24 h of rest (day 3). Rectal temperature was measured at the same time as blood sample collection. The majority of hematological and biochemical parameters remained within reference ranges at all sampling times. Rectal temperature was significantly higher after training on days 1 and 2 compared to resting temperature on day 3. Hematological parameters did not change significantly; however, there were significant differences in urea, creatinine, creatine kinase, and triglycerides between specific sampling times. Despite significant changes, these biochemical parameters remained within reference ranges. Significant changes in biochemical parameters seem to reflect the dogs' physiological response to each acute bout of exercise, considering all biochemical parameters and rectal temperature returned to pre-exercise values after a 24-h rest period (day 3). No significant differences in oxidative stress parameters were found between any sampling times. Relatively high erythrocyte superoxide dismutase1 activity at all sampling times may indicate that the canicross dogs are adapted to training by an increased expression of antioxidant enzymes. Based on our results, we can conclude that the trained canicross dogs included in our study were healthy, in good physical condition, and fit for the two acute bouts of field exercise.

Background and Objectives: Patients with schizophrenia are often exposed to polypharmacotherapy, which may lead to drug—drug interactions. The aim of the study was to investigate the prevalence of potential drug—drug interactions (pDDIs) in hospitalized patients with schizophrenia spectrum disorders and to identify factors associated with pDDIs and manifested symptoms and signs. Materials and Methods: This cross-sectional observational study included 311 inpatients admitted to a psychiatric hospital. The LexiComp drug interaction program was used to identify pDDIs in 2014. Factors associated with the prevalence of pDDIs and factors related to clinically observed symptoms and signs were assessed using multivariable regression. In addition, replicate analysis of pDDI was performed using 2021 program updates. Results: The prevalence of pDDIs was 88.7%. Our study showed that more than half of the patients received at least one drug combination that should be avoided. The most common pDDIs involved combinations of two antipsychotics or combinations of antipsychotics and benzodiazepines, which can lead to cardio-respiratory depression, sedation, arrhythmias, anticholinergic effects, and neuroleptic malignant syndrome. The number of prescribed drugs was a risk factor for pDDIs (OR 2.85; 95% CI 1.84–5.73). All groups of clinically observed symptoms and signs were associated with the number of drugs. In addition, symptoms and signs characteristic of the nervous system and psychiatric disorders were associated with antipsychotic dosage (IRR 1.33; 95% CI 1.12–1.58), which could contribute to the development of extrapyramidal syndrome, insomnia, anxiety, agitation, and bipolar mania. The 2021 version of the drug interaction program showed a shift in drug interactions toward a lower risk rating, implying less severe patient management and possibly less alert fatigue. Conclusions: Patients with schizophrenia spectrum disorders are at high risk of developing drug—drug interactions. Optimization of drug therapy, patient monitoring, and use of drug interaction programs could help to prevent pDDIs and subsequent adverse drug events.

The aim of this study was to investigate the effects of total intravenous anaesthesia with propofol and anaesthesia induced with propofol and maintained with sevoflurane on oxidative stress parameters in dogs with early-stage myxomatous mitral valve degeneration (MMVD). Sixteen client-owned dogs with early stage MMVD that required periodontal treatment were included in the study. After induction with propofol, anaesthesia was maintained with propofol (group P) or sevoflurane (group PS). Blood samples for determination of vitamin E, superoxide dismutase, glutathione peroxidase and malondialdehyde were collected before premedication, 5 and 60 minutes and 6 hours after induction to anaesthesia. There were no significant differences between groups in any of the oxidative stress parameters at each sampling time. Compared to basal values, vitamin E concentration decreased significantly during anaesthesia in both groups and glutathione peroxidase activity increased 60 minutes after induction to anaesthesia in PS group. Anaesthesia with propofol or with propofol and sevoflurane did not have any significant impact on oxidative stress parameters in dogs with early stage MMVD. In terms of oxidative stress, both protocols may be equally safely used in dogs with early stage MMVD.

Ustekinumab is a monoclonal antibody used in Crohn’s disease (CD). Dose optimization in case of non-response and the role of pharmacokinetic–pharmacodynamic (PK-PD) monitoring remain unresolved dilemmas in clinical practice. We aimed to develop a population PK-PD model for ustekinumab in CD and simulate efficacy of alternative dosing regimens. We included 57 patients and recorded their characteristics during 32 weeks after starting with ustekinumab therapy. Serum ustekinumab concentration was prospectively measured and fecal calprotectin (FC) concentration was used to monitor the disease activity. Ustekinumab PK-PD was described by a two-compartment target-mediated drug disposition model linked to an indirect response model. Lower fat-free mass, higher serum albumin, previous non-exposure to biologics, FCGR3A-158 V/V variant and lower C-reactive protein were associated with higher ustekinumab exposure. Model-based simulation suggested that 41.9% of patients receiving standard dosing achieve biochemical remission at week 32. In patients not achieving remission with standard dosing at week 16, transition to 4-weekly subcutaneous maintenance dosing with or without intravenous reinduction resulted in comparably higher remission rates at week 32 (51.1% vs. 49.2%, respectively). Our findings could be used to guide stratified ustekinumab treatment in CD, particularly in patients with unfavorable characteristics, who might benefit from early transition to 4-weekly maintenance dosing.

The effects of antioxidant supplements on exercise-induced oxidative stress have not been investigated in untrained leisure horses. We investigated the effects of 14-day supplementation with vitamin E (1.8 IU/kg/day), coenzyme Q10 (CoQ10; ubiquinone; 800 mg/day), and a combination of both (the same doses as in mono-supplementation) on the blood levels of CoQ10, vitamin E, and oxidative stress parameters in untrained leisure horses subjected to acute moderate exercise. Correlations between lipid peroxidation and muscle enzyme leakage were also determined. Forty client-owned horses were included in the study, with 10 horses in each of the antioxidant and placebo (paraffin oil) groups. Blood parameters were measured before supplementation, before and immediately after exercise, and after 24 h of rest. The differences in individual parameters between blood collection times and groups were analysed with linear mixed models (p ˂ 0.05). None of the supplemented antioxidants affected vitamin E and CoQ10 concentrations, oxidative stress parameters, or serum muscle enzymes. Lipid peroxidation occurred in horses supplemented with placebo and CoQ10 but not in horses supplemented with vitamin E or the combination of both antioxidants. These results suggest that vitamin E alone or in combination with CoQ10 prevented lipid peroxidation in untrained leisure horses subjected to acute moderate exercise.

Monoclonal antibodies (mAbs) have been extensively developed over the past few years, for the treatment of various inflammatory diseases. They are large molecules characterized by complex pharmacokinetic and pharmacodynamic properties. Therapeutic drug monitoring (TDM) is routinely implemented in the therapy with mAbs, to monitor patients’ treatment response and to further guide dose adjustments. Serum has been the matrix of choice in the TDM of mAbs and its sampling requires the visit of the patients to laboratories that are not always easily accessible. Therefore, dried blood spots (DBS) and various microsampling techniques have been suggested as an alternative. DBS is a sampling technique in which capillary blood is deposited on a special filter paper. It is a relatively simple procedure, and the patients can perform the home-sampling. The convenience it offers has enabled its use in the quantification of small-molecule drugs, whilst in the recent years, studies aimed to develop microsampling methods that will facilitate the TDM of mAbs. Nevertheless, hematocrit still remains an obstacle that hinders a more widespread implementation of DBS in clinical practice. The introduction of novel analytical techniques and contemporary microsampling devices can be considered the steppingstone to the attempts made addressing this issue.

Objective of the present report was to develop and validate a simple, sensitive, reliable, inexpensive and reproducible HPLC method with fluorescence detection (HPLC-FLD), suitable for routine therapeutic drug monitoring (TDM) of an antiepileptic drug topiramate. The determination of plasma topiramate concentration was carried out after precolumn derivatization, using 4-chloro-7-nitrobenzofurazan as a fluorescent labeling agent and bendroflumethiazide as an internal standard. The standard calibration curve was linear over the concentration range of 0.01-24 μg/mL (r²> 0.9998). The intra- and inter-day accuracies expressed as bias were from 1.41 to 9.95 % and from 1.85 to 10.23 %, respectively. The intra- and inter-day precisions were below 7.87 % and 2.73 %, respectively. A simple, sensitive, accurate, precise and inexpensive HPLC-FLD method for determination of topiramate in human plasma was developed and validated. The validated method was applied for the measurement of plasma topiramate concentrations in patients with epilepsy. The reported method is appropriate for TDM of topiramate as well as for pharmacokinetic and bioequivalence studies.

Purpose: The application of artificial neural networks in the pharmaceutical sciences is broad, ranging from drug discovery to clinical pharmacy. In this study, we explored the applicability of counter-propagation artificial neural networks (CPANNs), combined with genetic algorithm (GA) for prediction of topiramate (TPM) serum levels based on identified factors important for its prediction. Methods: The study was performed on 118 TPM measurements obtained from 78 adult epileptic patients. Patients were on stable TPM dosing regimen for at least 7 days; therefore, steady-state was assumed. TPM serum concentration was determined by high performance liquid chromatography with fluorescence detection. The influence of demographic, biochemical parameters and therapy characteristics of the patients on TPM levels were tested. Data analysis was performed by CPANNs. GA was used for optimal CPANN parameters, variable selection and adjustment of relative importance. Results: Data for training included 88 measured TPM concentrations, while remaining were used for validation. Among all factors tested, TPM dose, renal function (eGFR) and carbamazepine dose significantly influenced TPM level and their relative importance were 0.7500, 0.2813, 0.0625, respectively. Relative error and root mean squared relative error (%) and their corresponding 95% confidence intervals for training set were 2.14 [(-2.41) - 6.70] and 21.5 [18.5 - 24.1]; and for test set were -6.21 [(-21.2) - 8.77] and 39.9 [31.7 - 46.7], respectively. Conclusions: Statistical parameters showed acceptable predictive performance. Results indicate the feasibility of CPANNs combined with GA to predict TPM concentrations and to adjust relative importance of identified variability factors in population of adult epileptic patients. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

This article presents the results of mapping the Slovenian pharmacy curriculum to evaluate the adequacy of the recently developed and validated European Pharmacy Competences Framework (EPCF). The mapping was carried out and evaluated progressively by seven members of the teaching staff at the University of Ljubljana’s Faculty of Pharmacy. Consensus was achieved by using a two-round modified Delphi technique to evaluate the coverage of competences in the current curriculum. The preliminary results of the curriculum mapping showed that all of the competences as defined by the EPCF are covered in Ljubljana’s academic program. However, because most EPCF competences cover healthcare-oriented pharmacy practice, a lack of competences was observed for the drug development and production perspectives. Both of these perspectives are important because a pharmacist is (or should be) responsible for the entire process, from the development and production of medicines to pharmaceutical care in contact with patients. Nevertheless, Ljubljana’s graduates are employed in both of these pharmaceutical professions in comparable proportions. The Delphi study revealed that the majority of differences in scoring arise from different perspectives on the pharmacy profession (e.g., community, hospital, industrial, etc.). Nevertheless, it can be concluded that curriculum mapping using the EPCF is very useful for evaluating and recognizing weak and strong points of the curriculum. However, the competences of the framework should address various fields of the pharmacist’s profession in a more balanced way.

The growing threat of antibiotic resistance necessitates accurate differentiation between bacterial and viral infections for proper antibiotic administration. In this study, a Virus vs. Bacteria machine learning model was developed to distinguish between these infection types using 16 routine blood test results, C-reactive protein concentration (CRP), biological sex, and age. With a dataset of 44,120 cases from a single medical center, the model achieved an accuracy of 82.2 %, a sensitivity of 79.7 %, a specificity of 84.5 %, a Brier score of 0.129, and an area under the ROC curve (AUC) of 0.905, outperforming a CRP-based decision rule. Notably, the machine learning model enhanced accuracy within the CRP range of 10–40 mg/L, a range where CRP alone is less informative. These results highlight the advantage of integrating multiple blood parameters in diagnostics. The ''Virus vs. Bacteria'' model paves the way for advanced diagnostic tools, leveraging machine learning to optimize infection management.

Abstract Physicians taking care of patients with COVID-19 have described different changes in routine blood parameters. However, these changes hinder them from performing COVID-19 diagnoses. We constructed a machine learning model for COVID-19 diagnosis that was based and cross-validated on the routine blood tests of 5333 patients with various bacterial and viral infections, and 160 COVID-19-positive patients. We selected the operational ROC point at a sensitivity of 81.9% and a specificity of 97.9%. The cross-validated AUC was 0.97. The five most useful routine blood parameters for COVID-19 diagnosis according to the feature importance scoring of the XGBoost algorithm were: MCHC, eosinophil count, albumin, INR, and prothrombin activity percentage. t-SNE visualization showed that the blood parameters of the patients with a severe COVID-19 course are more like the parameters of a bacterial than a viral infection. The reported diagnostic accuracy is at least comparable and probably complementary to RT-PCR and chest CT studies. Patients with fever, cough, myalgia, and other symptoms can now have initial routine blood tests assessed by our diagnostic tool. All patients with a positive COVID-19 prediction would then undergo standard RT-PCR studies to confirm the diagnosis. We believe that our results represent a significant contribution to improvements in COVID-19 diagnosis.

The novel coronavirus SARS-CoV-2 can cause a severe and even fatal respiratory illness named COVID-19. Apart from respiratory failure, COVID-19 may be associated with various autoimmune complications. We present a case of a critically ill patient with COVID-19 who developed severe immune thrombocytopenia that was successfully treated with a concomitant use of corticosteroids and intravenous immunoglobulins.

Although anti-TBE vaccines are highly effective, vaccine breakthrough (VBT) cases have been reported. With increasing evidence for immune system involvement in TBE pathogenesis, we characterized the immune mediators reflecting innate and adaptive T and B cell responses in neurological and convalescent phase in VBT TBE patients. At the beginning of the neurological phase, VBT patients have significantly higher serum levels of several innate and adaptive inflammatory cytokines compared to healthy donors, reflecting a global inflammatory state. The majority of cytokines, particularly those associated with innate and Th1 responses, are highly concentrated in CSF and positively correlate with intrathecal immune cell counts, demonstrating the localization of Th1 and proinflammatory responses in CNS, the site of disease in TBE. Interestingly, compared to unvaccinated TBE patients, VBT TBE patients have significantly higher CSF levels of VEGF-A and IFN-β and higher systemic levels of neutrophil chemoattractants IL-8/CXCL8 and GROα/CXCL1 on admission. Moreover, serum levels of IL-8/CXCL8 and GROα/CXCL1 remain elevated for two months after the onset of neurological symptoms, indicating a prolonged systemic immune activation in VBT patients. These findings provide the first insights into the type of immune responses and their dynamics during TBE in VBT patients. An observed systemic upregulation of neutrophil derived inflammation in acute and convalescent phase of TBE together with highly expressed VEGF-A could contribute to BBB disruption that facilitates the entry of immune cells and supports the intrathecal localization of Th1 responses observed in VBT patients.