Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry最新文献

Cisplatin (CP) is an effective antineoplastic agent whose clinical application is constrained by organ toxicities, including liver injury. Although the underlying mechanisms of CP-associated hepatotoxicity have been widely explored, the characteristics of early biochemical and molecular responses following acute exposure remain incompletely defined. The present study aimed to examine early alterations in oxidative stress, inflammatory response, and apoptosis-related pathways following acute CP exposure and to evaluate their potential relevance for subsequent hepatotoxic progression. In this study, early alterations in oxidative balance, inflammatory mediators, and apoptosis-related proteins were examined in rat liver tissue after single-dose CP administration at 5, 7.5, and 12 mg/kg. Liver samples collected 3 days post-treatment were analyzed using ELISA-based assays, routine biochemical methods, and Western blotting for Bax and Bcl-2 expression. The results indicated a dose-related disruption of redox homeostasis, evidenced by elevated lipid peroxidation markers and reduced antioxidant activity. In parallel, pro-inflammatory cytokines showed an increasing trend across treatment groups. Apoptotic signaling was also affected, with enhanced Bax expression and reduced Bcl-2 levels, particularly at higher doses. These molecular findings were accompanied by increases in liver-associated enzyme activities. Overall, the data suggest that acute CP exposure induces measurable early changes at the biochemical and molecular levels in hepatic tissue. These alterations may represent initial responses associated with the development of hepatotoxicity. However, further studies incorporating extended observation periods and structural analyses are needed to better define the temporal progression and mechanistic relevance of these findings.

Insulin resistance (IR) is a critical metabolic hallmark of obesity that may significantly hinder weight loss, leading to a phenomenon of “weight stability.” This study aims to investigate the association between insulin resistance and weight stability in obese patients and to evaluate the clinical utility of IR-related biomarkers (insulin, HOMA-IR, C-peptide, and IGF-1) as predictive indicators of the body’s metabolic response to weight-loss interventions. A case-control study was conducted involving 80 participants: 40 obese patients (20 males, 20 females) and 40 healthy controls (20 males, 20 females), to ensure sufficient power to detect statistically significant differences whilst improving the robustness and reliability of the results. Furthermore, the sample size was considered appropriate in comparison with similar studies reported in the literature. Biomarkers were measured using chemiluminescence immunoassay (CLIA) on the Mindray CL-1000i system. Diagnostic performance, including sensitivity, specificity, and optimal cut-off values, was determined using receiver operating characteristic (ROC) curve analysis. Significant elevations (P < 0.001) in insulin, HOMA-IR, C-peptide, and IGF-1 were observed in obese patients compared to controls. ROC analysis demonstrated exceptional diagnostic accuracy for HOMA-IR and IGF-1, with an area under the curve (AUC) reaching 1.00 and 0.99, respectively, in female patients. Optimal cut-off values were established (e.g., 1.78 for HOMA-IR in males) to distinguish IR cases. Notably, while T.G. showed moderate diagnostic value, HDL exhibited poor sensitivity as a standalone marker. IGF-1 and HOMA-IR serve as highly precise biomarkers for identifying insulin resistance in obese individuals with weight stability. These findings suggest that metabolic profiling via CLIA technology can enhance early diagnosis and provide a biochemical explanation for the difficulty of weight loss, supporting personalized therapeutic strategies.

It was proved that adenosine deaminase (ADA) accumulates in synovial fluids (SFs) in rheumatoid arthritis (RA), reducing the anti-inflammatory adenosine concentration and exacerbating inflammation. Therefore, ADA inhibiting as an inflammation supporting agent may be beneficial in RA treatment. The quest for the new enzyme inhibitors is a relevant task. The aim of this work was to investigate the influence of the water and ethanol extracts of Origanum vulgare L. (OVL) from Armenian landscape on ADA activity. The influence of the extracts on ex vivo ADA activity in two groups of SFs from RA patients with differing the enzyme activity demonstrated more effective inhibition in SFs with higher ADA activity for both extracts. Two isoforms of ADA were purified to elucidate which one is the main factor responsible for the observed enzyme inhibition. There was no significant difference in inhibition of large isoform of ADA from human erythrocytes and RA SFs. The evaluated IC₅₀ values for small isoform of ADA (SADA) evidenced 1.5 times more effective inhibition in vitro citrullinated enzyme compared to the non-citrullinated one. Considering the accumulation of citrullinated SADA in RA SFs enhances inflammation by metabolizing adenosine, the obtained results suggest OVL is a promising source of anti-inflammatory agent for the development of new therapeutic medicines for RA.

Extracellular vesicles (EVs) represent a promising tool for diagnostics and therapy, but high heterogeneity limits their practical use. In this work, a comparative study of the physicochemical properties of native EVs secreted by HEK-293T cells and exosome-like nanovesicles (EMNVs) obtained by extrusion of whole HEK-293T cells was carried out. Using transmission electron microscopy, dynamic and electrophoretic light scattering, and surface-enhanced Raman spectroscopy, the authors found that EMNVs form a more homogeneous population with an average diameter of ~115 nm and a high negative charge (−22.9 mV), whereas native EVs are more heterogeneous in size and have a lower zeta potential (−12.1 mV). Spectral analysis revealed the presence of nucleic acid signals in native EVs and their absence in EMNVs, indicating differences in molecular composition due to different biogenesis mechanisms. The obtained data demonstrate that EMNVs are a more standardized and colloidally stable alternative to native EVs, making them a promising platform for the development of drug-delivery systems.

Women infertility is a common reproductive system disorder. Diagnosis and treatment are characterized by limited availability and high costs. This paper intends to evaluate the levels of endogenous antioxidant proteins metallothionein 2 (MT2) and metallothionein 4 (MT4) in the serum of both infertile and fertile women, as well as to assess the association between these proteins and total oxidant capacity (TOC), total antioxidant capacity (TAC), oxidative stress index (OSI), serum anti-Müllerian hormone (AMH), and serum testosterone. This case-control study involved one hundred eighty Iraqi women, categorized into three categories: primary infertile women (P group), secondary infertile women (S group), and healthy fertile women (C group). With regard to the control group, the results demonstrated a significant rise in serum MT2 levels within the primary infertile group, while serum MT4 level displayed a noticeable decrease in the infertile groups. The rise in MT2 could serve as a means to compensate for a decline in MT4, pointing out the potential antioxidant contribution of these proteins in the progression of infertility. A receiver operating characteristic (ROC) outcome demonstrated excellent sensitivity and specificity, alongside an elevated AUC for MT2, MT4, and other biomarkers in a diagnosis of women infertility.

Diabetic kidney disease is a serious complication of diabetes mellitus, often leading to chronic kidney disease and ultimately end-stage renal disease. Inflammation, besides deranged mineral metabolism, plays a role in the pathophysiology of diabetic kidney disease. The present study investigated the relationship between IL-6 an important pro-inflammatory cytokine, and Klotho an anti-aging protein with renoprotective function in diabetic kidney disease progression. A study was carried out on 60 patients with ESRD caused by diabetic kidney disease on maintenance hemodialysis and 40 apparently healthy control individuals. IL-6 and Klotho serum levels were measured using the enzyme-linked immunosorbent assay method. Some biochemical indicators, such as intact parathyroid hormone, 1.25-dihydroxyvitamin D₃, blood urea, and creatinine, were also measured. The main findings (mean ± SE) showed that the levels of IL-6 and iPTH levels were considerably higher in DKD patients (18.360 ± 2.323 pg/mL; 239.222 ± 24.073 pg/dL) than in controls (4.341 ± 0.393 pg/mL; 26.077 ± 1.609 pg/dL), indicating persistent inflammation and secondary hyperparathyroidism. Conversely, serum 1.25(OH)₂D₃ and Klotho levels were considerably lower in the DKD group (14.911 ± 0.991 ng/dL and 213.500 ± 25.905 pg/mL, respectively) than in controls (29.125 ± 1.195 ng/dL and 453.587 ± 33.797 pg/mL), indicating kidney function impairment and mineral metabolism disturbance. There was a notable weak inverse relationship between IL-6 and Klotho (r = –0.271, p = 0.036). This suggests that Klotho low levels may increase inflammatory responses and adversely affect renal function. In conclusion, the research demonstrates how IL-6 and Klotho affect one another in diabetic kidney disease. This suggests their application as potential biomarkers for disease progression and targets for therapy in preventing inflammation and maintaining kidney function.

This study evaluated the biochemical effects of high-intensity interval training (HIIT) protocols with different rest intervals on tryptophan–kynurenine pathway metabolites and trimethylamine N-oxide (TMAO) levels in young football players. Thirty healthy male amateur football athletes were randomly assigned to three groups: 20-s rest HIIT (n = 10), 30-s rest HIIT (n = 10), and a control group (n = 10). The HIIT groups performed the protocol twice weekly for six weeks in addition to routine football training. Serum samples were collected before and after the intervention, and concentrations of tryptophan, kynurenine, kynurenine pathway metabolites, TMAO, and related ratios were analyzed using LC-MS/MS. No significant differences in overall changes were observed between groups. However, within-group analyses revealed protocol-specific responses. In the 20-s rest group, tryptophan and kynurenic acid decreased, while 3-hydroxykynurenic acid increased; the KYN/TRP and KYNA/KYN ratios also changed significantly. In the 30-s rest group, quinolinic acid and the QA/KYN ratio decreased, whereas the 3-HAA/3-HK ratio increased. No significant changes were observed in the control group, and TMAO levels remained unchanged across all groups. These results suggest that rest interval duration can modulate kynurenine pathway metabolism in a protocol-specific manner, highlighting the importance of considering recovery parameters when designing exercise interventions targeting neurometabolic biomarkers.

Background. Subclinical hypothyroidism (SCH) may increase cardiovascular risk through early alterations in inflammation (hs-CRP), atherogenicity (AIP), and glycemic status (HbA1c). This study aimed to evaluate the association and discriminatory ability of thyroid-stimulating hormone (TSH) with these cardiovascular risk markers in individuals with subclinical hypothyroidism (SCH). Methods. A comparative case-control study was performed with 76 patients diagnosed with SCH and 72 healthy controls matched for age and sex. Anthropometric and biochemical parameters were evaluated, including thyroid hormones (TSH, FT3, FT4), anti-thyroid peroxidase (anti-TPO) antibodies, lipid profile, hs-CRP, AIP, fasting blood glucose (FBG), and HbA1c. Correlations between thyroid markers and cardiovascular risk indicators (hs-CRP, AIP, HbA1c) were examined using Spearman’s correlation, binary logistic regression, and receiver operating characteristic (ROC) curve analysis. Results. SCH patients exhibited markedly elevated levels of total cholesterol, triglycerides, LDL, VLDL, and AIP compared with controls (p < 0.05), whereas HDL, fasting blood glucose, and HbA1c showed no significant differences. Although hs-CRP levels did not differ significantly between groups (p = 0.061), TSH was positively correlated with hs-CRP (r = 0.343, p = 0.002) and AIP (r = 0.235, p = 0.041). Multiple regression analysis identified TSH as an independent determinant of hs-CRP after adjustment for potential confounding variables. ROC analysis demonstrated improved discriminatory performance for hs-CRP after adjustment, with the AUC increasing from 0.747 to 0.838 (ΔAUC = 0.0906, p = 0.0227), supporting the association of TSH with inflammatory status in SCH. Conclusions. In SCH, elevated TSH is linked to the early inflammatory and atherogenic changes indicated by hs-CRP and AIP. These findings support the use of TSH as a simple and cost-effective indicator for cardiovascular risk stratification in SCH. Regular monitoring of hs-CRP and AIP may enable the early identification of high-risk patients, while forthcoming prospective trials should evaluate whether early normalization of TSH can reduce inflammation and enhance cardiovascular outcomes.

Proteases represent a rather heterogeneous group of enzymes that accomplish hydrolysis in peptide bond, and are central in a wide range of biological mechanisms, be-coming a major focus of medical research. Proteases can now be traditionally known as involved in the process of the protein degradation but today they have been known to regulate many physiological activities, including proliferation, apoptosis, immune amendment, and tissue reshaping. Protease expression or activity aberrations, particularly those that occur during the development and progression of many diseases including cancer, cardiovascular diseases, inflammatory diseases, and neurodegenerative syndromes are closely correlated to the levels of disease pathogenesis. Consequently, these enzymes have received a high level of interest both as biomarkers of early disease diagnoses as well as treatment targets. Particular attention is paid to natural and artificial proteases in the current clinical practice or under development as therapeutic agents in several treatment modes, such as thrombolytic and wound debridement, cancer therapy, and drug delivery. Integrating the existing research, the paper includes the most important discoveries and areas that still need to be under-stood, making the research comprehensive and resting on the fact that proteases are inherently the catalytic tools of nature that have the power to change the way things are used in the medical world today.

The production of recombinant proteins requires careful optimization of all bioprocess stages, starting from the selection of the expression system and cultivation conditions to the control of critical process parameters (CPPs) and critical quality attributes (CQAs). Current regulatory requirements (ICH Q8(R2), ICH Q9, FDA, EMA, EAEU) are focused on the implementation of the quality by design (QbD) concept, which ensures a systematic approach to development and quality management. One of the key tools of QbD is the use of design of experiments (DoE), which allows for the identification and optimization of parameters influencing protein yield and quality. This study discusses the application of DoE in the development of recombinant protein cultivation processes in E. coli, including proinsulin aspart and glargine and two peptides with molecular masses of up to 5 kDa. It is demonstrated that the use of DoE enables the establishment of relationships between CPPs and CQAs, reduces process variability, increases outcome predictability, and decreases development timelines by at least twofold. The obtained data confirm the effectiveness of integrating statistical methods into bioprocess development, contributing to enhanced standardization, reproducibility, and compliance with international quality standards.