Krzysztof Safranow

Background/Objectives: In this study, we investigated whether targeting thiamine (vitamin B₁) metabolism could be useful in the treatment of glioblastoma. Methods: The study was conducted using LN-229 and T98G glioblastoma cell lines, as well as glioblastoma tissue samples obtained from up to 30 patients. The effects of thiamine and the thiamine antimetabolites oxythiamine and amprolium on cell viability were assessed using the MTT assay. The effects of hypoxia and temozolomide (TMZ), as well as gene expression in tissue samples, were analyzed using qRT-PCR. The expression of SLC19A2, SLC19A3, TPK1, SLC25A19, SLC35F3, and SLC44A4 was evaluated. Results: Higher thiamine concentrations increased the MTT metabolic activity of glioblastoma cells. Oxythiamine and amprolium reduced MTT metabolic activity, although their inhibitory effects reached a plateau and neither compound enhanced the effect of TMZ. Chronic hypoxia reduces thiamine metabolism. The expression of proteins involved in thiamine metabolism is lower in glioblastoma tissue than in peritumoral tissue. Conclusions: Overall, targeting thiamine metabolism does not appear to improve the efficacy of TMZ-based glioblastoma treatment. However, the observed viability-promoting effect of higher thiamine concentrations indicates that excessive thiamine intake or supplementation in patients with glioblastoma should be approached with caution and requires further clinical investigation.

Objectives: To characterise longitudinal structural and functional retinal restoration after inverted internal limiting membrane (ILM) flap surgery for idiopathic full-thickness macular holes (FTMHs), with a particular emphasis on postoperative glial proliferation, outer retinal restoration, and structure–function relationships. Methods: This retrospective consecutive case series included 46 eyes of 44 patients with idiopathic FTMH treated using the inverted ILM flap technique. Spectral-domain optical coherence tomography (SD-OCT), best-corrected visual acuity (BCVA), microperimetry, fixation stability, and multifocal electroretinography (mfERG) were performed 1, 6, and 12 months postoperatively. Results: Anatomical closure was achieved in all eyes. Complete restoration of both the ELM and EZ increased from 8.7% at 1 month to 45.7% at 6 months and 54.3% at 12 months, whereas severe glial proliferation decreased from 30.4% to 15.2% at 12 months. BCVA, retinal sensitivity, fixation stability, and central mfERG responses improved predominantly between 1 and 6 months postoperatively. In multivariable analysis, lower closure-pattern grade (B = −0.073, 95% CI: −0.141 to −0.005; p = 0.035) and lower glial proliferation grade (B = −0.091, 95% CI: −0.155 to −0.027; p = 0.006) were independently associated with better BCVA at final visit. Conclusions: Retinal healing after inverted ILM flap surgery is a coordinated, time-dependent process involving regression of postoperative gliosis, restoration of the outer retinal microstructure, and multimodal functional recovery. Longitudinal integration of OCT with microperimetry, fixation analysis, and mfERG provides a comprehensive framework for understanding postoperative retinal remodelling beyond anatomical hole closure.

Rheumatoid arthritis (RA) is an autoimmune disease with a complex pathogenesis. It has been shown that epigenetic modifications can influence the differentiation and function of many immune system cells involved in the pathogenesis of RA. Important epigenetic factors include non-coding RNAs, such as long non-coding RNAs (lncRNAs). Non-coding RNAs can influence numerous immune processes in RA. This study aimed to evaluate the expression of lncRNAs SNHG7, SNHG12, and SNHG14 in the synovial membranes of patients with RA and to correlate these results with selected clinical parameters of the disease. This study included 46 patients diagnosed with RA and 67 healthy individuals as the control group. The expression levels of the lncRNAs SNHG7, SNHG12, and SNHG14 in synovial membranes in patients with RA were statistically higher than those in the control group. The synovial expression of SNHG14 showed a statistically significant negative correlation with C-reactive protein values. There were no statistically significant correlations between synovial expression of SNHG7, SNHG12, and clinical parameters. The study results suggest that lncRNAs may potentially play a role in the development of RA.

Background: The tryptophan catabolic pathway may play an important role in maternal–fetal immune tolerance, placental function, and metabolic programming, yet its clinical correlates in uncomplicated term pregnancies are not well defined. This study examined how maternal and umbilical artery blood tryptophan metabolites relate to maternal characteristics in women undergoing cesarean section. Methods: In this cross-sectional study, 73 women delivering by cesarean section at a mean gestational age of 38.9 ± 0.96 weeks were enrolled. Maternal venous and umbilical cord arterial blood were collected at delivery, and serum tryptophan (TRP), kynurenine (KYN), kynurenic acid (KYNA), and quinolinic acid (QUIN) were measured using ELISA. Maternal anthropometry, pregnancy complications, antibiotic treatment, and indications for cesarean section were obtained from medical records. Group comparisons used Mann–Whitney U tests, associations were assessed with Spearman’s rank correlations, and selected outcomes were analyzed in multivariable linear regression models. Results: Umbilical concentrations of TRP, KYN, KYNA, and QUIN were higher than the corresponding maternal levels. Maternal TRP was inversely associated with gestational age at delivery and was independently higher in women treated with antibiotics during pregnancy. Cord KYNA correlated positively with pre-pregnancy and pre-delivery BMI, with a stronger association for postpartum BMI, and remained independently associated with pre-delivery BMI in multivariable analysis. Maternal QUIN was higher in women undergoing intrapartum or emergency cesarean section and in those delivered by cesarean section for breech presentation, and both indications were independently associated with higher QUIN in the regression model. No robust associations were observed between TRP, KYN, KYNA, or QUIN and most pregnancy complications, neonatal birth weight, or Apgar scores. Conclusions: Maternal and fetal tryptophan–kynurenine metabolites show distinct, domain-specific relationships with gestational timing, maternal antibiotic exposure during pregnancy, adiposity, and indications or mode of cesarean delivery in term pregnancies. Maternal TRP levels may primarily reflect gestational age and concurrent inflammatory status. Cord blood KYNA may mirror maternal metabolic condition, whereas maternal QUIN appears to be more closely associated with complex obstetric phenotypes than with neonatal status. These exploratory, uncorrected findings suggest that kynurenine pathway metabolites may be associated with selected maternal anthropometric and obstetric factors; none of these associations survived correction for multiple comparisons, and they are strictly hypothesis-generating, requiring confirmation in larger, prospectively designed studies before any biomarker application can be considered. The parallel elevation of the cord KYN/TRP and KYNA/KYN ratios, together with a relative reduction in the QUIN/KYN ratio in the fetal compartment, suggests a metabolic profile characterized by preferential conversion of kynurenine toward the KYNA branch rather than the QUIN branch. This pattern may be consistent with a more neuroprotective metabolic environment in the fetal circulation; however, mechanistic studies are required to confirm this interpretation. The maternal QUIN should be regarded as a marker of obstetric complexity rather than a direct consequence of delivery mode. The study included only women undergoing cesarean delivery; therefore, the findings cannot be generalized to vaginal deliveries.

In hemodialysis patients, Body Mass Index is insufficient in assessing their nutritional status due to the ‘obesity paradox’ and the association between body composition and inflammation. This study assessed the relationship between body composition, traditional inflammatory markers, the new NETosis indicators (neutrophil extracellular traps), and their association with 12-month mortality. The study included 99 maintenance hemodialysis (HD) patients. Their body composition was assessed using bioelectrical impedance analysis. Blood serum was tested for inflammatory markers (hs-CRP-high sensitive c-reactive protein, IL-6 interleukin-6, TNF-α tumor necrosis factor alfa, IL-1β interleukin-1 beta), NETosis markers (citrullinated histone CH3, myeloperoxidase -MPO, elastase), and nutritional status parameters (albumin, transferrin). No correlation between BMI -body mas index and inflammation was demonstrated. Higher adipose tissue, particularly visceral, was significantly positively correlated with IL-6 and hs-CRP levels, while muscle mass was negatively correlated with inflammation. Dialysis catheter use was associated with higher CH3 levels (NETosis indicator) and lower albumin concentrations. Low albumin levels and high TNF-α levels were independent predictors of death. Body composition, rather than BMI, is associated with the severity of inflammation. Visceral obesity is positively correlated with increased inflammation, while muscle mass shows an inverse association. Dialysis catheters are linked to higher NETosis markers and lower albumin levels, which are associated with a poorer prognosis.

Introduction: In hemodialysis patients, Body Mass Index is insufficient in assessing their nutritional status due to the ‘obesity paradox’ and the impact of body composition on inflammation. The aim of the study was to assess the relationship between body composition, traditional inflammatory markers, and the new NETosis indicators (neutrophil extracellular traps), as well as to determine their impact on 12-month mortality. Methods: The study included 99 patients with end-stage renal disease. Their body composition was assessed using bioelectrical impedance analysis (Seca mBCA 525). Blood serum was tested for inflammatory markers (hs-CRP, IL-6, TNF-α, IL-1B), NETosis markers (citrullinated histone H3, MPO, elastase), and nutritional status parameters (albumin, transferrin). Results: No correlation between BMI and inflammation was demonstrated. Higher contents of the adipose tissue, particularly visceral, were significantly associated with increased levels of IL-6 and hs-CRP, while muscle mass was negatively correlated with inflammation. The use of dialysis catheters stimulated NETosis (higher CH3 levels), which had a negative effect on albumin concentrations. Low albumin levels and high TNF-α levels were independent predictors of death. Conclusions: It is body composition, and not BMI, that determines the severity of inflammation. Visceral obesity promotes inflammation, while muscle mass has a protective effect. Dialysis catheters, by stimulating NETosis, contribute to a decrease in albumin levels and a poorer prognosis.

The treatment of chronic kidney disease is based mainly on haemodialysis. During this procedure, cardiovascular haemodynamic parameters change, which can lead to complications in the circulatory system and shorten patient survival. Currently, new biomarkers are being sought that could predict survival in haemodialysis patients. The aim of this study was to evaluate whether Gal-3, GDF-15, sST2, and CysC could be useful biomarkers in patients with end-stage renal failure undergoing haemodialysis in predicting the patient survival. This study included 76 patients with end-stage kidney disease treated with haemodialysis. We assessed the variability in the plasma concentrations of these proteins; evaluated correlations between Gal-3, GDF-15, and sST2 concentrations and selected haemodialysis parameters; and determined the relationship between the risk of death and Gal-3, GDF-15, sST2, and CysC concentrations. There was a decrease in the Gal-3 plasma concentrations and an increase in the GDF-15 and sST2 plasma concentrations during haemodialysis. Among the studied proteins, only the sST2 concentrations differed significantly between the patients who survived and died. The logarithm of the sST2 concentrations, both before and after haemodialysis, was an independent mortality risk factor in univariate analysis (respectively: hazard ratio [HR] 3.179, 95% confidence interval [CI] 1.492–6.777, p = 0.0027, and HR 3.011, 95% CI 1.447–6.264, p = 0.0032) and multivariate analysis, which also included age, male gender, and body mass index (respectively: HR 2.776, 95% CI 1.306–5.903, p = 0.0080, and HR 2.496, 95% CI 1.224–5.090, p = 0.012). These results suggest the use of sST2 in clinical practice to identify patients at higher risk of death.

Although combination therapies with mitogen activated protein kinase inhibitors remain among the most effective treatments for malignant melanoma, they are not universally applicable to all subtypes of this cancer, and their efficacy decreases in the presence of distant metastases. Drug resistance, often associated with elevated autophagy in tumor cells, and adverse effects of the treatment also reduce the survival time of melanoma patients. Therefore, the aim of our research was to assess the cytotoxicity of the combination of a late-stage autophagy blocker chloroquine with thymoquinone, a natural substance with anticancer potential and low toxicity towards healthy cells, in metastatic melanoma cell lines. Using the WST-1 assay, we examined the cytotoxicity of the combination of chloroquine and thymoquinone in melanoma WM9 and WM852 cell lines and assessed the type of their interactions. Additionally, using time-lapse bright field and fluorescent microscopy, we assessed changes in cell morphology during 48 h incubation with the tested compounds and their combinations, as well as their effect on autophagy. We identified an additive and sub-additive cytotoxic effect of thymoquinone/chloroquine combinations against WM9 and WM852 cells. Moreover, we found that thymoquinone combined with chloroquine caused an increase in autophagosome accumulation in WM9 cells, while attenuating the chloroquine’s anti-autophagic effect. A thorough understanding of the mechanism of drug interactions with natural substances is crucial for the development of new effective anticancer therapies.

The development of drug resistance remains a major obstacle in the treatment of metastatic melanoma, necessitating novel therapeutic strategies. Combining natural compounds with agents that disrupt cancer cell survival mechanisms, such as autophagy inhibitors, offers a promising approach. This study aimed to evaluate the potential synergy between the plant-derived monoterpene carvacrol and the autophagy inhibitor chloroquine against metastatic melanoma. The cytotoxic and proapoptotic effects of carvacrol, chloroquine, and their combination were investigated in the WM9 metastatic melanoma cell line using WST-1 assays and Annexin V/7-AAD flow cytometry. The nature of the drug interaction was quantified using the Q-value. In silico molecular docking was performed to identify potential protein targets and elucidate the mechanisms underlying the observed effects. While carvacrol monotherapy exhibited weak cytotoxicity, its combination with non-toxic concentrations of chloroquine resulted in a potent and synergistic reduction in WM9 cell viability. This enhanced cytotoxicity was attributed to a significant, synergistic induction of apoptosis. In silico analysis predicted that both carvacrol and chloroquine bind with high affinity to common molecular targets, including Insulin-Like Growth Factor 1 Receptor and Sirtuin-2. Chloroquine was also predicted to interact strongly with HSP90. Our findings demonstrate that chloroquine potentiates the anticancer activity of carvacrol in metastatic melanoma cells. The synergy observed is likely mediated by a multi-targeted mechanism involving the simultaneous disruption of key cancer survival and signaling pathways. This study establishes the carvacrol-chloroquine combination as a novel and effective strategy that warrants further preclinical investigation for the treatment of resistant melanoma.

We aimed to validate the feasibility of combining ultra-widefield (UWF) fundus photography with targeted swept-source optical coherence tomography (SS-OCT) for clinical decision-making regarding a prophylactic laser therapy. For this purpose we enrolled 119 patients (135 eyes) who, basis on fundus examination, were eligible for prophylactic photocoagulation of degenerative retinal lesions. Eyes were classified into two groups: (1) justified laser, when SS-OCT confirmed vitreoretinal traction and/or subretinal fluid beneath the neurosensory retina; and (2) non-justified laser, when SS-OCT did not confirm these criteria. Using this SS-OCT-guided UWF approach, we found that 25.1% of eyes that initially qualified for laser based on clinical examination did not meet the SS-OCT criteria. Patients in the justified laser group were significantly younger than those in the non-justified group. Horseshoe retinal tears, lattice degeneration and snail-track degenerations, multiple lesions, and lesions located in the far and mid-periphery were significantly more frequent in the justified laser group than in the non-justified group. By contrast, the prevalence of operculated holes, bilateral lesions, and degenerative lesions in patients with a retinal detachment in the fellow eye did not differ between groups. Our findings suggest the SS-OCT-guided UWF imaging may refine patient selection for prophylactic laser therapy.