Pathophysiology, Volume 32, Issue 2 (June 2025) – 9 articles

Background/Objectives: The pathogenesis of intensive care unit-acquired weakness (ICU-AW) is multi-factorial, with some of the main risk factors being sepsis, multiorgan failure, and the inflammatory response related to critical illness. Vitamin D is crucial for muscle function, the immune response, and inflammation, and has been identified as a predictor of negative outcomes in intensive care unit (ICU) patients with COVID-19. The objective of this preliminary study was to examine the relationship between vitamin D serum levels and the incidence of ICU-AW in a cohort from the University Hospital of Split. Methods: A prospective observational cohort study was conducted in the University Hospital of Split in ICU from December 2021 to March 2022. The inclusion criteria were as follows: patients over 18 years old who had a confirmed severe acute respiratory coronavirus disease 2 (SARS-CoV-2) infection, patients who were mechanically ventilated for more than 48 h, and patients who were weaned from a ventilator over at least 24 h. The exclusion criteria were a history of neurological or musculoskeletal disorders and a pre-existing poor functional status. Vitamin D was detected in the first routine blood sample. Results: A total of 77 patients were observed, with 36 patients who were successfully weaned from a ventilator over at least 24 h and 1 patient who could not be examined because of impaired consciousness (this patient was excluded from further analysis), and thus a total of 35 patients were analyzed. Of these 35 patients, 12 (34%) developed ICU-AW. The median vitamin D serum level in the ICU-AW group was 17 (7.5–73.3), while that in the non-ICU-AW group was 25.2 (12.3–121). The difference in vitamin D serum levels between the groups was not significantly different from zero (p = 0.567). All patients, except for one, were vitamin D insufficient. Conclusions: Vitamin D serum levels in the ICU-AW group were not statistically different from the non-ICU-AW group, possibly due to the small sample size. Given the known roles of vitamin D in muscle function, immune modulation, and inflammation, a potential etiopathogenetic role in ICU-AW cannot be excluded without additional studies. Therefore, further studies with larger sample sizes than ours are necessary to determine whether vitamin D deficiency contributes to the development of ICU-AW and whether supplementation could have preventive or therapeutic value. Full article

Background/Objectives: This study explores variations in brain activity between individuals with dementia of the Alzheimer’s type (DAT) and healthy older adults during a resting state using functional near-infrared spectroscopy (fNIRS). Methods: FNIRS measured brain activity in ten AD patients and six healthy individuals. A device with 16 channels was placed on each participant’s forehead to measure oxygenation levels while they kept their eyes closed. The data were analyzed using a support vector machine (SVM) model. Results: The results indicated differences in oxygenated hemoglobin (HbO) levels between the two groups. Specifically, HbO levels were generally higher in the dementia group in the left hemisphere, with a sharp increase after 26 s. Conversely, HbO levels were consistently lower in the right hemisphere of the dementia group. The SVM analysis demonstrated high accuracy in differentiating between the AD and healthy groups based on HbO levels. Conclusions: The study indicates that differences in brain activity during resting state can potentially distinguish people with DAT from healthy individuals. We found relatively reduced hemoglobin activity in the prefrontal areas of those with DAT. Furthermore, the concentration changes in the HbO in the left lateral prefrontal and right medial brain regions emerged as the most informative in distinguishing individuals with DAT from healthy individuals. The results of the current study show that this method could improve current DAT diagnostic practices due to its efficiency. Full article

Background: Podocytes are essential for kidney function, and their dysfunction can result in nephrotic syndrome, such as minimal change disease (MCD). Oxidative stress contributes to podocyte damage. We investigated the therapeutic potential of intravenously infused mesenchymal stem cells (MSCs) in a puromycin aminonucleoside (PAN)-induced rodent MCD model, focusing on oxidative stress modulation. Methods: Sprague-Dawley rats were divided into three groups: intact, PAN-Vehicle, and PAN-MSC. MCD was induced through subcutaneous PAN injection. MSCs were infused intravenously in the PAN-MSC group on day 7. Urinary albumin, serum albumin, and creatinine levels were assessed. Histological analysis of the renal cortex was performed. Podocyte protein (NPHS1, NPHS2, and PODXL) and antioxidant enzyme (SOD1, SOD2, and GPX1) levels were measured using quantitative real-time reverse-transcription PCR (qRT-PCR). Results: MSC infusion significantly reduced proteinuria and restored podocyte structure in the PAN-MSC group. Electron microscopy revealed that infused MSCs could inhibit the fusion of the foot process induced by PAN injection. qRT-PCR showed that intravenous infusion of MSCs rescued the inhibition of GPX1 expression. GFP-labeled MSCs accumulated at the podocyte injury sites. Conclusion: Systemic MSC infusion mitigates PAN-induced MCD by reducing proteinuria, preserving podocyte structure, and modulating oxidative stress via the GPX1 pathway, offering a potential therapeutic approach for nephrotic syndrome. Full article

Background: Pregnancy simulates a metabolic syndrome-like state and predisposes to iodine deficiency and hypothyroidism through increased iodine renal loss and transplacental transfer to the fetus. Iodine deficiency is thought to predispose to dyslipidemia through elevation of serum TSH. Obesity, dyslipidemia, and hypothyroidism are established risk factors of preeclampsia. Hence, pregnant women with iodine deficiency are likely to be at increased risk of dyslipidemia and preeclampsia. We investigated the pattern of dyslipidemia among preeclamptic and normotensive pregnant women with and without iodine deficiency. Methods: The pathophysiological mechanisms linking iodine deficiency and dyslipidemia were delineated using bivariate correlations, logistic regression, and exploratory factor analysis of anthropometric, lipid profile, urine iodine concentration (UIC), and thyroid function data from 240 women with preeclampsia and 120 normotensive pregnant controls at term who attended Lomo Medical Centre, Democratic Republic of Congo (DRC). Results: Preeclamptic women with iodine deficiency had significantly lower HDL-C but higher triglyceride levels than those with sufficient iodine intake. Both normotensive and preeclamptic participants with elevated TSH had high serum oxidized LDL-C but low NO, p < 0.001. Conclusions: SCH, secondary to iodine deficiency, is associated with elevated serum oxidized LDL and decreased Nitric Oxide (NO) among both normotensive and preeclamptic women, while insufficient iodine nutrition among preeclamptic women predisposes to reduced HDL-C and increased serum Triglycerides, which are risk factors of atherosclerosis and cardiovascular disease. Full article

Currently, understanding the immune response, its abnormalities, and its diagnostic possibilities is a key point in the management of patients with various diseases, from infectious to oncological ones. The aim of this review was to analyze the data presented in the current literature on immune disorders and the possibility of their laboratory diagnostics in combination with clinical manifestations. We have performed a systematic analysis of the literature presented in international databases over the last ten years. We have presented data on the possibility of diagnosing immunopathological processes due to changes in immune cells and soluble molecules involved in the pathogenesis of a wide range of diseases, as well as the determination of antibodies to detect autoimmune processes. By applying laboratory techniques such as hematology, flow cytometry, ELISA, etc., available to most clinical laboratories worldwide, clinical data on immune system dysfunction in a wide range of diseases are being collected. This process is unfortunately still very far from being completed. However, with all the diversity of accumulated knowledge, we can currently state that the pathogenesis of the vast majority of immune-mediated diseases is not yet known. At the same time, the current success in dividing immune-mediated diseases into distinct clusters based on different types of inflammatory responses that are based on the involvement of different populations of T helper cells and cytokine molecules represents significant progress. Further research in this direction seems very promising, as it allows the identification of new target cells and target molecules for both improved diagnostics and targeted therapies. Full article

Background: Metabolic effects of oleoylethanolamide-based dietary supplement (OEA-DS) were studied in a model of dietary-induced obesity in mice. Obesity was induced by a 2-month high-fat, high-cholesterol diet, resulting in significant morphological changes in liver tissues and elevated cholesterol levels in the animals’ blood serum. Elevated levels of proinflammatory cytokines, oxidative stress, and hepatocyte apoptosis were also observed in the liver tissue. The aim of this study was to examine the mechanisms through which an OEA-based dietary supplement (OEA-DS) exerts a comprehensive influence on multiple aspects of the pathogenesis of MASLD, thereby demonstrating a robust hepatoprotective effect. Methods: mice were fed a high-fat, high-cholesterol diet with or without OEA-DS supplementation. Liver tissues and blood serum were analyzed for cholesterol levels, inflammatory markers (CD68, Iba-1, CD163, IL-1β, IL-6, TNFα), apoptotic markers (Bad, Bax, Bcl-2), nuclear receptors (PPAR-α, PPAR-γ, AdipoR1), and enzymes involved in lipolysis (Acox1, Cpt1a) and cholesterol metabolism (Ldlr, Furin, Pcsk9). Immunohistochemistry, Western blotting, and RT-PCR were used to assess protein expression and gene transcription. Results: administration of OEA-DS normalized cholesterol levels, decreased expression of inflammatory markers (CD68 and Iba-1), pro-apoptotic markers (Bad, Bax) and levels of pro-inflammatory cytokines (IL-1β, IL-6, TNFα). In parallel, the expression of nuclear receptors PPAR-α and PPAR-γ, adiponectin receptor 1 (AdipoR1), and anti-inflammatory (CD163) and anti-apoptotic (Bcl-2) markers have risen. OEA-DS administration induced the expression of liver lipolysis enzymes (Acox1, Cpt1a) and cholesterol metabolism factors (Ldlr, Furin), while simultaneously reducing the transcription of the proatherogenic factor Pcsk9. Conclusions: The results of this study suggest a complex action of OEA-DS in obesity-associated liver damage, which includes reduction of systemic inflammation. Full article

Background/Objectives: For patients with medically refractory temporal lobe epilepsy (TLE), surgery is an effective strategy. However, post-operative seizure recurrence occurs in 20–30% of patients, and it remains challenging to predict outcomes solely based on clinical variables. Here, we ask to what extent differences in gene expression in epileptic tissue can predict the outcome after resective epilepsy surgery. Methods: We performed RNAseq on hippocampal tissue resected from eight patients who underwent anterior temporal lobectomy with amygalohippocampectomy (ATL/AH), half of whom became seizure free (SF) or non-seizure free (NSF). Results: Bioinformatic analyses revealed 1548 differentially expressed genes and statistical enrichment analyses identified a distinct set of pathways in NSF and SF cohorts that were associated with neuroinflammation, neurotransmission, synaptic plasticity, and extracellular matrix (ECM) reorganization. Resected tissue exhibiting strong pro-inflammatory processes are associated with better post-surgery seizure outcomes than patients exhibiting cellular signaling processes related to ECM reorganization, autoantibody production, and neural circuit formation. Conclusions: The results suggest that post-operative targeting of both inhibitory aspects of the ECM remodeling and the autoimmune/inflammatory components may be helpful in promoting repair and preventing the recurrence of seizures. Full article

Background/Objectives: Two of the microvascular complications in type 2 diabetes (T2D) are diabetic retinopathy (DR), which is the most common cause of non-traumatic blindness, and diabetic kidney disease (DKD); the latter generally requires renal replacement therapy. The aim of the present study was to determine the frequency of polymorphisms of Tumor Necrosis Factor-α, interleukin-6, and interleukin-10 (TNF-α, IL-10, and IL-6), as well as to describe the clinical and laboratory characteristics of T2D association with these microvascular complications. Methods: This study included 203 patients with T2D, of which 102 had microvascular complications: 95 with DR, 50 with DKD, and 15 with diabetic neuropathy (the latter were not included in the statistical analysis); those with T2D without confirmed microvascular complications were considered as controls. Clinical and laboratory data were collected from the patient’s medical records. Polymorphism typing of TNF-α rs361525 and rs1800629 and IL-10 rs1800872 and rs1800871 were obtained using MALDI-TOF MS. IL-10 rs1800896 and IL-6 rs1800795 were typed using a quantitative real-time polymerase chain reaction. Results: The results of age, HbA1c, fasting glucose, and arterial hypertension are significantly associated in every group. The TNF-α rs1800629A allele and TNF-α rs1800629G/A genotype were associated with microvascular complications and DR. For IL-10-rs1800896, all the models were associated in DKD. The TNF-α rs361525-rs1800629GA haplotype was associated with microvascular complications and DR, while the IL-10 haplotype, rs1800872-rs1800871-rs1800896 GGC, showed susceptibility in every group. Conclusions: Our results show the contributions of the variants of these cytokines to these microvascular complications, but more studies are required to reach relevant conclusions. Full article

Background/Objectives: Proprotein convertase subtilisin/kexin type 9 (PCSK9) regulates serum cholesterol levels and inflammation, both of which are dysregulated in inflammatory bowel disease (IBD). Free cholesterol (FC) and the various types of cholesteryl ester (CE) have different functions in the body. However, it is not yet known whether these lipids undergo parallel changes in male and female patients with active IBD, nor whether PCSK9 correlates with these lipids and disease severity in either sex. The present study measured the serum levels of PCSK9, FC, and 15 CE species in IBD patients, focusing on the associations of these molecules with sex, each other, and with disease severity. Methods: The serum PCSK9 levels of 80 IBD patients (42 males and 38 females) and 24 controls (12 males and 12 females) were measured by enzyme-linked immunosorbent assay. In addition, FC and 15 CE species levels of 53 randomly selected IBD patients and 16 controls were determined by direct flow injection analysis (FIA) using a high-resolution hybrid quadrupole-orbitrap mass spectrometer (FIA-FTMS). Results: Serum PCSK9 levels in controls and IBD patients were comparable and did not correlate with disease severity in IBD patients. There was no discernible difference in serum PCSK9, FC, and CE levels between patients with Crohn’s disease (CD) and those with ulcerative colitis (UC). FC and almost all CE species decreased in male patients with active IBD but were not related to disease severity in the female patients. The decrease in different CE species in male IBD patients with diarrhea compared to those with normal stool consistency appears to be related to IBD severity. Bile acids regulate serum cholesterol levels, and FC and CE levels were positively correlated with fecal levels of secondary bile acids in the patients with UC but not CD. This association also existed in male UC patients and could not be evaluated in women due to the small sample size. Conclusions: In active IBD, a reduction in FC and almost all CE species was observed only in males, while serum PCSK9 levels remained within normal ranges in both sexes. It can be hypothesized that blocking PCSK9 may further reduce serum cholesterol levels, which may have adverse effects in male patients with active IBD. Full article