In the past, several animal disease models were developed to study the molecular mechanism of neurological diseases and discover new therapies, but the lack of equivalent animal models has minimized the success rate. A number of critical issues remain unresolved, such as high costs for developing animal models, ethical issues, and lack of resemblance with human disease. Due to poor initial screening and assessment of the molecules, more than 90% of drugs fail during the final step of the human clinical trial. To overcome these limitations, a new approach has been developed based on induced pluripotent stem cells (iPSCs). The discovery of iPSCs has provided a new roadmap for clinical translation research and regeneration therapy. In this article, we discuss the potential role of patient-derived iPSCs in neurological diseases and their contribution to scientific and clinical research for developing disease models and for developing a roadmap for future medicine. The contribution of humaniPSCs in the most common neurodegenerative diseases (e.g., Parkinson’s disease and Alzheimer’s disease, diabetic neuropathy, stroke, and spinal cord injury) were examined and ranked as per their published literature on PUBMED. We have observed that Parkinson’s disease scored highest, followed by Alzheimer’s disease. Furthermore, we also explored recent advancements in the field of personalized medicine, such as the patient-on-a-chip concept, where iPSCs can be grown on 3D matrices inside microfluidic devices to create an in vitro disease model for personalized medicine.

Background: Scientific studies point to a significant global vitamin D deficiency. The recommended dose of vitamin D for the adult population in Central Europe is 800–2000 IU/day. The aim of our study was to determine whether doses of 1000 IU or 2000 IU of vitamin D₃ are adequate to achieve the sufficiency reference values of [25(OH)D]. Methods: Seventy-two healthy volunteers, average age twenty-two, took part in the study. The study was conducted from October to March in order to eliminate intra-dermal vitamin D production. Vitamin D₃ in an oleaginous mixture was used. The participants used either 1000 IU or 2000 IU/daily for two 60-day periods with a 30-day break. Results: The dose of 1000 IU, taken for 60 days, increased vitamin D levels relatively little. Furthermore, serum vitamin D levels decreased in the 30 days following the cessation of supplementation. Taking 2000 IU daily led to a sharp increase in serum levels which plateaued 30 days after the subjects stopped using vitamin D₃ drops. Conclusions: Both doses, taken daily, can help maintain adequate vitamin D levels during the winter months. A daily dose of 2000 IU, however, maintained the desired levels of vitamin D for a longer period.

The glycoprofiling of two proteins, the free form of the prostate-specific antigen (fPSA) and zinc-α-2-glycoprotein (ZA2G), was assessed to determine their suitability as prostate cancer (PCa) biomarkers. The glycoprofiling of proteins was performed by analysing changes in the glycan composition on fPSA and ZA2G using lectins (proteins that recognise glycans, i.e. complex carbohydrates). The specific glycoprofiling of the proteins was performed using magnetic beads (MBs) modified with horseradish peroxidase (HRP) and antibodies that selectively enriched fPSA or ZA2G from human serum samples. Subsequently, the antibody-captured glycoproteins were incubated on lectin-coated ELISA plates. In addition, a novel glycoprotein standard (GPS) was used to normalise the assay. The glycoprofiling of fPSA and ZA2G was performed in human serum samples obtained from men undergoing a prostate biopsy after an elevated serum PSA, and prostate cancer patients with or without prior therapy. The results are presented in the form of an ROC (Receiver Operating Curve). A DCA (Decision Curve Analysis) to evaluate the clinical performance and net benefit of fPSA glycan-based biomarkers was also performed. While the glycoprofiling of ZA2G showed little promise as a potential PCa biomarker, the glycoprofiling of fPSA would appear to have significant clinical potential. Hence, the GIA (Glycobiopsy ImmunoAssay) test integrates the glycoprofiling of fPSA (i.e. two glycan forms of fPSA). The GIA test could be used for early diagnoses of PCa (AUC = 0.83; n = 559 samples) with a potential for use in therapy-monitoring (AUC = 0.90; n = 176 samples). Moreover, the analysis of a subset of serum samples (n = 215) revealed that the GIA test (AUC = 0.81) outperformed the PHI (Prostate Health Index) test (AUC = 0.69) in discriminating between men with prostate cancer and those with benign serum PSA elevation.

Sesquiterpene lactones are secondary plant metabolites with sundry biological effects. In plants, they are synthesized, among others, for pesticidal and antimicrobial effects. Two such compounds, archangelolide and trilobolide of the guaianolide type, are structurally similar to the well-known and clinically tested lactone thapsigargin. While trilobolide has already been studied by us and others, there are only scarce reports on the biological activity of archangelolide. Here we present the preparation of its fluorescent derivative based on a dansyl moiety using azide–alkyne Huisgen cycloaddition having obtained the two sesquiterpene lactones from the seeds of Laserpitium archangelica Wulfen using supercritical CO₂ extraction. We show that dansyl-archangelolide localizes in the endoplasmic reticulum of living cells similarly to trilobolide; localization in mitochondria was also detected. This led us to a more detailed study of the anticancer potential of archangelolide. Interestingly, we found that neither archangelolide nor its dansyl conjugate did exhibit cytotoxic effects in contrast to the structurally closely related counterparts trilobolide and thapsigargin. We explain this observation by a molecular dynamics simulation, in which, in contrast to trilobolide, archangelolide did not bind into the sarco/endoplasmic reticular calcium ATPase cavity utilized by thapsigargin. Last, but not least, archangelolide exhibited anti-inflammatory activity, which makes it promising compound for medicinal purposes.

Background: The medication used to treat benign prostate hyperplasia (BPH), a common condition in men over 50 years of age, can alter the levels of biomarkers used in prostate cancer detection. Commonly used medications for BPH include alpha‐blockers, 5‐alpha reductase inhibitors (5‐ARIs), and muscarinic antagonists. We studied the impact of these drugs on total prostate‐specific antigen (tPSA), free PSA (fPSA), [−2]proPSA, fPSA/tPSA ratio, and the Prostate Health Index (PHI), as well as novel potential biomarkers in the form of glycan composition of fPSA.Patients and Methods: Serum samples were collected from 564 males with BPH, with a mean age of 68.5 years. The samples were used to measure levels of tPSA, fPSA, and [−2]proPSA. The fPSA/tPSA and PHI were then calculated. The glycan composition of fPSA was analyzed using lectin‐based glycoprofiling. Pharmacotherapy data was collected from the patients' medical records.Results: Alpha‐blocker monotherapy was associated with higher fPSA and fPSA/tPSA ratio, and decreased PHI. Levels of tPSA were not impacted. Alpha‐blocker and 5‐ARI dual therapy was associated with reduced levels of fPSA, [−2]proPSA, and PHI. Therapy combining alpha‐blockers and antimuscarinic agents did not significantly influence biomarker levels apart from an increase in a Maackia amurensis lectin‐recognized glycan originating in fPSA.Conclusion: BPH pharmacotherapy notably affects prostate cancer biomarkers. Recognizing the impact of pharmacotherapy is crucial for achieving an accurate diagnosis of prostate cancer and for planning treatment.

Accurate assessment of vitamin D metabolism is crucial not only for the diagnosis and treatment of disorders related to bone health and calcium homeostasis but also for understanding its broader physiological roles in immunity, cellular differentiation, cardiovascular regulation, and endocrine function. Although 25‐hydroxyvitamin D ₃ (25(OH)D ₃ ) is routinely measured in clinical practice, the low‐abundance metabolite 24,25‐dihydroxyvitamin D ₃ (24,25(OH) ₂ D ₃ ) provides complementary insight into vitamin D catabolism. Reduced or undetectable 24,25(OH) ₂ D ₃ levels may signal impaired CYP24A1 function or insufficient conversion of 25(OH)D ₃ to maintain appropriate intracellular concentrations of biologically active 1,25(OH) ₂ D ₃ . However, quantification of 24,25(OH) ₂ D ₃ remains analytically challenging and requires highly selective and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) methodology. In‐house developed LC–MS/MS approaches are being increasingly employed, offering greater specificity and sensitivity over conventional immunoassays, allowing accurate detection of low‐abundance vitamin D metabolites. Additionally, when interpreted together with 25(OH)D ₃ , 24,25(OH) ₂ D ₃ allows the calculation of the vitamin D metabolite ratio (VMR), which offers a more accurate assessment of vitamin D sufficiency and catabolism. The aim of this study was to develop and validate a robust LC–MS/MS method using dynamic multiple reaction monitoring (dMRM) and sample derivatization for quantifying 24,25(OH) ₂ D ₃ in human serum. The method described was evaluated according to EMA guidelines and DEQAS controls. The matrix effect was minimized through lipid‐removal filtration. The assay demonstrated excellent linearity ( R ² = 0.9982), intra‐ and inter‐assay precision below 14%, and LOQ of 0.64 ng/mL. Recovery from DEQAS samples ranged from 80% to 118%, with a matrix‐induced ion enhancement of ∼17%. Additionally, 4‐phenyl‐1,2,4‐triazoline‐3,5‐dione (PTAD) derivatization enhanced the sensitivity 100‐fold. This highly sensitive LC–MS/MS method is suitable for clinical and research laboratories equipped with an electrospray ionization (ESI) source. Precise quantification of 24,25(OH) ₂ D ₃ can complement routine 25(OH)D ₃ analysis, support VMR determination, and serve as a reliable biomarker for disorders associated with altered vitamin D metabolism.