Cellular Physiology and Biochemistry最新文献

Background/aims: Contaminants in the environment pose a considerable threat to biodiversity, ecological balance, and the health of both wildlife and humans, particularly through the transfer of these harmful substances via fish in the food chain.

Methods: This study focused on the developmental stages of sea trout (Salmo trutta L.) in both riverine and Baltic Sea environments, with the aim of exploring how chemical element accumulation influences oxidative stress biomarkers in these species.

Results: The findings revealed notable age- and tissue-specific patterns in the accumulation of chemical elements in sea trout. Specifically, higher levels of lead (Pb), arsenic (As), mercury (Hg), and tin (Sn) were detected in the muscle tissues of adult trout, while cadmium (Cd) primarily accumulated in the gills, particularly in smolts. These results underscore the influence of both age and tissue type on the bioaccumulation of contaminants in the trout, highlighting how the accumulation of toxic elements contributes to increased oxidative stress in the fish. This oxidative stress, reflected by increased lipid peroxidation (TBARS) and carbonyl derivatives of oxidatively modified proteins, was closely related to the presence of contaminants such as Cd, Pb, As, Hg, and Sn. Gills, which are directly exposed to waterborne pollutants, exhibited significantly higher levels of oxidative damage compared to muscle tissue, consistent with the greater accumulation of metals in this organ. Despite higher total antioxidant status (TAS) in muscle tissue, both muscle and gill tissues of adult trout showed signs of considerable oxidative stress, indicating the cumulative effects of prolonged exposure to these contaminants.

Conclusion: The study highlights the detrimental consequences of chemical element contamination on the health of trout, with a particular emphasis on oxidative damage, and calls for effective environmental management to protect aquatic species from the long-term effects of exposure to contaminants. Furthermore, the correlation and regression analysis conducted revealed significant patterns, demonstrating positive correlations between the accumulation of Cd, Pb, and As in the gills of adult trout, and between Pb and oxidative stress markers in smolts. Additionally, the analysis indicated that mercury contributes significantly to oxidative damage.

Background/aims: Methylglyoxal (MG) is associated with the development of metabolic disorders that modify the hepatic energetic metabolism in different ways. However, not much is known about the effects of MG on energy metabolism in healthy liver cells. Therefore, this study investigated the effects of daily MG administration to Wistar rats on hepatic and fat tissue energetic metabolism.

Methods: Rats received MG intraperitoneally at doses of 100 or 200 mg/kg for seven consecutive days in acute approach or at a dose of 25 mg/kg for one month in the chronic approach. Metabolic pathways were measured in isolated perfused livers (glycogen catabolism, gluconeogenesis and ketogenesis) as well in adipose tissue. Activities and mRNA expressions of gluconeogenic enzymes were assessed in the liver and the viability of human lymphocytes were evaluated in vitro.

Results: MG displayed systemic inflammation and the metabolic changes were similar to those of widespread catabolic diseases. MG and advanced glycation end-products stimulated lymphocyte proliferation, and MG increased the hepatic interleukin-6 expression. Rats that received MG developed insulin resistance. Gluconeogenesis was diminished and glycolysis was stimulated in livers of rats that received MG. Two factors contribute to this outcome: a deficiency in mitochondrial energy supply and a much more significant downregulation of gluconeogenic enzymes. The adipose tissue metabolism was modified in a way that the AMPK-induced lipolysis was increased in the retroperitoneal fat, but not in the mesenteric fat. Ketogenesis was increased and triglycerides content was decreased in the liver.

Conclusion: To what degree the modifications in hepatic metabolism found in MG-exposed rats can be translated to patients with a high-grade inflammation and cirrhosis is uncertain. However, it is unlikely that the strong catabolic state induced by MG would not contribute in some way to the hepatic dysfunction in advanced liver diseases.

Background/aims: Pregnancy is associated with changes in renal hemodynamics, such as increases in renal blood flow and the glomerular filtration rate (GFR). Angiotensin-converting enzyme 2 (ACE2), a transmembrane glycoprotein involved in vasodilation, also acts as a receptor for the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during coronavirus disease 2019 (COVID-19).

Methods: Using rats on pregnancy day 16 and postpartum day 5, we examined the histopathological changes in rat kidneys during and after pregnancy. The expressional changes in renal angiotensin-converting enzyme 2 (ACE2) and angiotensin (1-7) (Ang (1-7)) were examined, together with those of transmembrane protease serine 2 (TMPRSS2).

Results: examined, together with those of transmembrane protease serine 2 (TMPRSS2). Results: In pregnant rats, the renal arterioles and venules as well as the glomerular capillaries were markedly dilated, indicating renal vasodilation. Immunohistochemistry demonstrated increased ACE2 and Ang (1-7) expression within the proximal renal tubules during pregnancy, which then returned to the virgin levels in the postpartum period. Additionally, the proximal tubular expression of ACE2 and TMPRSS2 was similarly enhanced during pregnancy.

Conclusion: As ACE2 and Ang (1-7) exert vasodilatory properties, they were considered responsible for renal vasodilation and the subsequent increase in GFR. Further, the similar distribution and enhanced expression of ACE2 and TMPRSS2 in the proximal renal tubules during pregnancy suggested their roles in the development of acute kidney injury (AKI) following COVID-19 in pregnancy. This study highlights the physiological and pathological significance of ACE2 during pregnancy.

Background/aims: This research explores the effectiveness of a new cytotoxic T-cell epitope peptide specific for HLA-A2402 in enhancing cellular immune responses to SARS-CoV-2 infections. HLA molecules play a key role in presenting antigenic epitopes to T cells, with genetic polymorphisms resulting in varying immune responses among individuals. The study aimed to investigate whether loading this epitope peptide into dendritic cells (DCs) from HLA-A2402(+) individuals could improve immune responses.

Methods: DCs were sensitized with varying doses of the peptide (2-12 µg/mL), with optimal results observed at 8 µg/mL. T-cell responses, proliferation, differentiation, Th cytokine secretion, CTL function, and apoptotic response were compared among the HLA-A2402(+), HLA-A2402(-), PBS control, DC only, and epitope-only groups.

Results: A significant enhancement in DC maturation, antigen presentation, T-cell activation, and proliferation was observed in the HLA-A2402(+) group compared to the HLA-A2402(-) control.

Conclusion: These findings suggest that HLA-A2402-restricted epitope peptides can enhance cellular immunity, offering potential for improving allele-specific SARS-CoV-2 vaccines and other molecular therapies, advancing precision medicine for infectious diseases.

Background/aims: Sudden Sensorineural Hearing Loss (SSNHL) is a rapid-onset condition with varied etiologies, creating uncertainty in optimal treatment strategies. This study aimed to assess whether adding pentoxifylline to standard antiviral and steroid therapy could enhance hearing recovery in SSNHL patients.

Methods: This randomized controlled trial was conducted between January 2021 and June 2023 in a private clinic in Chennai district, Tamil Nadu. Seventy-two patients aged 20-70 years with SSNHL onset within 7 days were randomized to receive either standard therapy (antiviral and steroids) or additional pentoxifylline. Serial pure-tone audiometry was performed to evaluate hearing recovery.

Results: The intervention group showed significantly improved hearing recovery, particularly when treatment was initiated within 72 hours of symptom onset. Non-responders underwent MRI of the brain, revealing cerebellopontine angle pathology in selected cases. Statistical analysis confirmed superior outcomes with pentoxifylline add-on therapy (p < 0.05).

Conclusion: Pentoxifylline may enhance hearing recovery when combined with antiviral and steroid therapy in SSNHL, especially when administered early. Intratympanic dexamethasone and MRI are recommended for non-responders.

Background/aims: In recent years, significant progress has been made in treating cartilage lesions, leading to various techniques aimed at articular surface regeneration. However, the impact of patient age on treatment outcomes remains understudied, despite its recognized influence on effectiveness. Many studies impose upper age limits, resulting in limited data on middle-aged patients, a group frequently undergoing cartilage repair. Age-related physiological changes, including a decline in regenerative capacity and alterations in proteoglycan composition, are believed to affect treatment success. This study investigates the clinical outcomes of cartilage repair in patients aged 45 to 60, specifically evaluating the effectiveness of microfracture combined with Hyalofast membrane implantation.

Methods: Patients aged 45-60 undergoing microfracture with Hyalofast membrane implantation were evaluated. Short- and medium-term outcomes were assessed using the KOOS and SF-36 questionnaires preoperatively and at 6- and 12-month follow-ups. Functional evaluations and MRI analyses were conducted by expert reviewers. Additionally, an intensive rehabilitation program was initiated immediately post-surgery. Statistical analysis was performed using a one-way repeated-measures ANOVA to compare preoperative, 6-month, and 12-month results, with Tukey's post-hoc test applied for multiple comparisons. Clinical significance was assessed using Cohen's d effect size, with standard thresholds for small, medium, and large effects.

Results: The combined microfracture and Hyalofast membrane approach, along with early rehabilitation, yielded promising results. Key benefits included reduced inflammation, improved membrane integration, and decreased subchondral edema.

Conclusion: These findings support the feasibility of this treatment strategy, highlighting its potential to enhance joint function, alleviate pain, and improve the overall quality of life in middle-aged patients.

Background/aims: Acinetobacter baumannii is a ubiquitous opportunistic microorganism associated with high morbidity and mortality Particularly among burn patients and immunocompromised individuals. This study aimed to assess the time-dependent antimicrobial effectiveness of indolicidin in combination with tigecycline against multidrug-resistant (MDR) A. baumannii isolates from patients with wound infections.

Methods: The antimicrobial synergy between indolicidin and tigecycline was evaluated using the checkerboard broth microdilution method And anti-biofilm activity was assessed using the crystal violet assay. Ten isolates resistant to multiple antibiotics were identified and confirmed using the API 20-NE system.

Results: Fractional inhibitory concentration (FIC) values ranged from 0.97 to 250 µg/ml. The results demonstrated that the inhibitory activity of indolicidin and tigecycline was significantly enhanced when combined With FIC values between 0.046 and 0.312 µg/ml. The MIC of indolicidin for isolate A4 decreased from 62.5 to 0.97 µg/ml • While the MIC of tigecycline decreased from 500 to 15.6 µg/ml • Indicating a strong synergistic effect. Additionally The combination therapy led to a 78% reduction in biofilm formation.

Conclusion: This study suggests a novel combination therapy for treating wound infections caused by MDR A. baumannii • Demonstrating significant efficacy in both bacterial inhibition and biofilm reduction.

Background/aims: Anthropogenic impact is irreversibly changing natural habitats of birds. Changes caused by the bioaccumulation of trace metals can lead to the development of oxidative stress and affect oxygen-dependent metabolic pathways in bird tissues, which can be used as effective bioindicators in these conditions. The objectives of our study were (a) to investigate the tissue-specific activity of key enzymes involved in metabolic changes and energy production, including Krebs cycle enzymes, as well as variations in metabolites associated with oxygen-dependent processes; and (b) to apply multivariate regression analysis, using beta and correlation coefficients, to elucidate the mechanisms of adaptive responses in pigeons to environmental changes in lead-contaminated areas.

Methods: This study investigates the ecotoxicological effects on feral pigeons (Columba livia f. urbana) in their natural habitats. It examines the influence of key environmental factors, sex, and biochemical alterations across five tissues (liver, kidney, heart, muscle, and brain). The analysis includes the combined effects of these variables on biochemical biomarkers related to energy metabolism, oxidative stress, and antioxidant defenses, considering the levels of chemical elements present in the pigeons. The analyses involved two groups of pigeons, namely, 7 females and 10 males (n = 17) in the group sampled in Słupsk and 7 females and 7 males, (n = 14) in Szpęgawa that living in two areas in central part of Northern Poland, which differed in the level of anthropopressure.

Results: We report significant values of lead bioaccumulation in feathers of pigeons and the impact of this metal on the activities of Krebs cycle enzymes (succinate dehydrogenase, pyruvate dehydrogenase, isocitrate dehydrogenase, α-ketoglutarate dehydrogenase), biomarkers of oxygen-dependent processes (lactate dehydrogenase activity, lactate and pyruvate levels, and their ratio), and aminotransferases in different tissues of pigeons.

Conclusion: Biomarkers of oxygen-dependent processes in five tissues of pigeons are depending on sex and environment. Pigeons from lead-exposed areas exhibited decreased antioxidant defence by biochemical alterations in tissues. Analytical model of oxidative stress biomarkers, Krebs cycle enzymes, and chemical elements is significant. Using multivariate regression analysis with beta- and correlative coefficients, relationships were shown for the optimal development of adaptation alterations in biochemical reactions in pigeons in response to the modification of their environments. Research on Columba livia f. urbana provides valuable insights into understanding the effects of anthropogenic pollution on bird physiology and offers practical methods for assessing environmental health.

Background/aims: DNA isolation is the initial process in genetic research. The product is used in many PCR reactions (PCR-RFLP, Real-Time PCR, multiplex PCR). That is why it is important to optimize DNA isolation protocol to obtain a good quality of DNA. Our first attempts at isolation, conducted using Purification Kit, did not result in sufficient concentration (6.414 ng*μL^-1) and purity (A-260/280) of 0.764 of isolated DNA.

Methods: We used twice the recommended amount of tissue and cell lysis solution to get more effective cell lysis. We extend the time of vortexing, centrifugation and incubation at critical steps. We manipulated the speed and temperatures of centrifugation. We used cold iso-propanol to get white strands of DNA faster. When rinsing with ethanol we used cold alcohol. We tested efficiency of two methods of drying of ethanol to achieve optimal DNA pureness. We leave the isolated DNA for 20 minutes to evaporate the ethanol and then resuspend nucleic acid in TE Buffer.

Results: Our modifications resulted in the improvement of isolation efficiency. After optimization we achieved DNA concentration (in range of 50-150 ng*μL^-1) and purity (A 260/280) of 1.735. Similar results for DNA parameters were achieved from the whole blood frozen for 2-3 months (concentration in the range of 125.762 ng*μL^-1, pureness: 1.761) and from blood frozen for 18 months (117.94 ng*μL^-1 and 1.7194, respectively). We performed electrophoresis after each isolation to confirm the effectiveness of optimized procedure. The refinements we used in DNA isolation are more efficient than those recommended in DNA Purification Kits.

Conclusion: Our results confirm that optimized DNA protocol fulfills the conditions of good extraction technique: it is relatively fast and easy to perform yet it guarantees a high reproducibility, specificity and sensitivity. There are also no dangerous or harmful steps. Our paper demonstrates innovative and effective approach. It confirms a high effectiveness of method regardless of duration of sample freezing, as well as introduce important modifications (timing, temperature conditions, drying details, absence of K-proteinase) that make overall procedure more productive and relatively fast.