Search Results (32,142)

Background: The rising rates of gallstones and cholecystectomy in pediatric populations underscore the increasing concern regarding chronic cholecystitis. However, the morphopathogenesis of pediatric calculous cholecystitis is still not well understood. This study aimed to determine the expression and distribution of immunomodulatory factors interleukin-12 (IL-12), interleukin-13 (IL-13), interleukin-1β (IL-1β), sonic hedgehog protein (SHH), nuclear factor NF-kappa-B p65 subunit (NFkBp65), and heat shock protein 60 (HSP60) in the gallbladder walls of pediatric patients with chronic calculous cholecystitis. Methods: In total, 11 gallbladder samples were collected from pediatric patients with calculous cholecystitis during cholecystectomy, while 5 healthy gallbladder samples served as controls. IL-12, IL-13, IL-1β, SHH, NFkBp65, and HSP60 were detected by immunohistochemistry. The number of positive structures in gallbladder wall epithelium, vasculature, and inflammatory infiltrate was assessed semi-quantitatively by microscopy. A Mann–Whitney U test and Spearman’s rank-order correlation coefficient were calculated. Results: Statistically significant differences were observed between patient and control samples in the expression of IL-1β, SHH, and NFkBp65 in the epithelium, as well as in the expression of IL-12, SHH, and HSP60 in the blood vessels. The expression of IL-1β was stronger in the epithelium of controls, while other markers were more prominent in patient samples. Conclusions: An increased number of NFkBp65, IL-12, and HSP60 positive cells in patient gallbladder tissue suggests a significant role of these tissue factors in driving immune modulation and sustaining the inflammation in pediatric chronic calculous cholecystitis. The noticeable expression of SHH in patient gallbladder tissue indicates its part in tissue regeneration and repair processes, as well as in modulating inflammation and vascular responses in calculous cholecystitis. The significant positive correlations between the factors studied highlight the importance of their coordinated interaction and intricate crosstalk in the morphopathogenesis of calculous cholecystitis. Full article

Enterotoxigenic Escherichia coli (ETEC) produces two types of enterotoxins, LTs and STs, as well as several colonization factors (CFs), including CS21, CS3 fimbriae, and flagellar structures. This study investigated how these structures contribute to ETEC colonization and the immune response in HT-29 and HuTu-80 intestinal cells. ETEC strains with single, double, and triple mutations in the lngA, cstH, and fliC genes were generated and confirmed using PCR and Western blotting. The colonization of HT-29 and HuTu-80 intestinal cells by the ETEC E9034A strain, which was fully sequenced using a hybrid approach involving both Illumina and Oxford Nanopore technologies, was used to generate the mutant and recombinant proteins. The colonization and adherence of E9034A and its mutants were assessed through colony-forming unit (CFU) counts. Cytokine levels were assessed using flow cytometry and analyzed via FlowJo 7.6.1. Quantitative analysis revealed that the absence of the lngA, cstH, and fliC genes significantly (p < 0.01) reduced ETEC adherence to HT-29 and HutU-80 cells. In addition, only ETEC strains expressing the FliC protein induced IL-8 secretion. These findings suggest that LngA, CstH, and FliC in ETEC E9034A enhance adherence to intestinal cells and trigger the release of IL-8. Full article

Background: Excessive accumulation of glomerular extracellular matrix (ECM) is a key factor in the development and progression of diabetic nephropathy (DN). As kidney dysfunction has been reported in normoalbuminuric patients, identifying novel diagnostic and prognostic markers is essential for the prevention and treatment of DN. Methods: Urinary excretion of neutrophil gelatinase-associated lipocalin (NGAL) and ECM-related glycoproteins, i.e., fibronectin (FN) and laminin (LN), was measured in obese patients with newly diagnosed type 2 diabetes mellitus (T2DM) before and after 6 months of metformin therapy. Results: Baseline NGAL (1.27 (0.80–2.36) ng/mg Cr), FN (11.19 (5.31–21.56) ng/mg Cr) and LN (123.17 (54.56–419.28) pg/mg Cr) levels did not significantly differ between T2DM patients and controls (1.95 (1.09–2.97) ng/mg Cr, 11.94 (7.78–18.01) ng/mg Cr and 157.85 (83.75–326.40) pg/mg Cr, respectively). In multivariate regression analysis, the body mass index was identified as the only significant predictor influencing urinary NGAL and FN levels at baseline, with β = 0.249, p = 0.005 and β = 1.068, p = 0.010, respectively. Metformin treatment significantly increased urinary levels of both ECM proteins, i.e., FN (18.48 (11.64–32.46) ng/mg Cr) and LN (179.51 (106.22–414.68) pg/mg Cr), without any effect on NGAL levels (1.44 (0.81–2.72) ng/mg Cr). FN and LN were positively associated with NGAL both before (r = 0.709 and r = 0.646, both p < 0.001, respectively) and after (r = 0.594 and r = 0.479, both p < 0.001, respectively) therapy. No correlations were found between NGAL, FN, LN, and albuminuria. However, NGAL was positively correlated with the albumin/creatinine ratio (ACR) both before (r = 0.323, p < 0.05) and after (r = 0.287, p < 0.05) therapy, and negatively with estimated glomerular filtration rate (eGFR) in pre-treatment diabetics (r = −0.290, p < 0.05). FN and LN were also correlated with ACR (r = 0.384, p < 0.01 and r = 0.470, p < 0.001), although the association for LN was limited to untreated patients (r = 0.422, p < 0.01). Conclusions: Our results suggest that metformin has a beneficial effect on ECM turnover with a significant increase in urinary excretion of non-collagenous markers of glomerular injury, i.e., FN and LN. Additionally, ECM-related markers may serve as useful tools for monitoring early renal injury in obese diabetic patients. Full article

The use of insecticides is widespread in the control of debilitating mosquito-borne diseases. P450 enzymes (CYPs) play essential roles in mosquito physiological function but also in the enzymatic detoxification of xenobiotics. Broadly speaking, CYPs can be classified as “stable”, meaning those that have no or very few paralogs, and “labile”, constituting gene families with many paralogous members. The evolutionary dichotomy between “stable” and “labile” P450 genes is fuzzy and there is not a clear phylogenetic demarcation between P450s involved in detoxification and P450s involved in essential metabolic processes. In this study, bioinformatic methods were used to explore differences in the sequences of “stable” and “labile” P450s that may facilitate their functional classification. Genomic and sequence data of Anopheles gambiae (Agam), Aedes aegypti (Aaeg), and Culex quinquefasciatus (Cqui) CYPs were obtained from public databases. The results of this study show that “stable” CYPs are encoded by longer genes, have longer introns and more exons, and contain a higher proportion of hydrophobic amino acids than “labile” CYPs. Compared to “labile” CYPs, a significantly higher proportion of “stable” CYPs are associated with biosynthetic and developmental processes. Full article

Phallus impudicus is a fungus used as a medicine and nutrient-rich food. However, the shelf life of mature Phallus impudicus is only a few hours. Therefore, research on its preservation technology is essential for improving its economic value. This study investigated the effects of hexanal concentrations (25–100 μL/L) and treatment time (4–8 h) on the inhibition of peach-shaped Phallus impudicus (CK) maturation and found that the maturation rate was 25% under optimal conditions of 25 μL/L hexanal treatment for 6 h. Quantitative transcriptomic and lipidomic analyses were conducted among CK, mature Phallus impudicus (M-P), and hexanal-treated peach-shaped Phallus impudicus (H-P-P). In total, 2933 and 2746 differentially expressed genes (DEGs) and 156 and 111 differentially abundant lipids (DALs) were identified in CK vs. H-P-P and M-P vs. H-P-P, respectively. Functional analysis demonstrated that hexanal treatment inhibited phospholipase D gene expression and reduced phosphatidic acid abundance, thereby inhibiting the activation of the phosphatidylinositol signaling system and the signal amplification of the cell wall integrity mitogen-activated protein kinase pathway. These blocked signal transductions inhibited the gene expression of most β-glucanases, chitinases and chitin synthases, further affecting cell wall reconstruction. Moreover, hexanal treatment enhanced membrane stability by reducing the monogalactosyl diglyceride/digalactosyl diacylglycerol ratio and increasing the phosphatidylcholine/phosphatidylethanolamine ratio. This study contributed to the development of hexanal treatment as a postharvest preservation technology for Phallus impudicus. Full article

Sulfonylureas (SUs)—a class of drugs primarily used to treat type 2 diabetes—have recently attracted interest for their potential anticancer properties. While some studies have explored the chemical modification or design of new SU derivatives, our work instead centers on biological evaluations of all commercially available SUs in combination with doxorubicin (DOXO). These antidiabetic agents act by stimulating insulin secretion via K_ATP channel inhibition, and because K_ATP channels share structural features with ATP-binding cassette (ABC) transporters involved in multidrug resistance (e.g., P-glycoprotein, MRP1, and MRP2), SUs may also reduce cancer cell drug efflux. In this study, we systematically examined each commercially available SU for potential synergy with DOXO in a panel of human cancer cell lines. Notably, combining DOXO with glimepiride (GLIM), the newest SU, results in a 4.4-fold increase in cytotoxicity against MCF-7 breast cancer cells relative to DOXO alone. Mechanistic studies suggest that the observed synergy may arise from increased intracellular accumulation of DOXO. Preliminary in vivo experiments support these findings, showing that DOXO (5 mg/kg, i.v.) plus GLIM (4 mg/kg, i.p.) is more effective at inhibiting 4T1 tumor growth in mice than DOXO alone. Additionally, we show that adding a small amount of the surfactant Tween-80 to culture media affects SU binding to bovine serum albumin (BSA), potentially unmasking anticancer effects of SUs that strongly bind to proteins. Overall, these results underscore the potential of repurposing existing SUs to enhance standard chemotherapy regimens. Full article

Passiflora edulis Sims peel (Chinese name Baixiangguo, BXG) is a by-product with a high nutritional and economic value of Passiflora edulis Sims. In this study, corn was partly replaced with BXG to make feed for finishing pigs and the effects on the carcass traits, meat quality, muscle amino acid profile, and gene expression of finishing pigs were evaluated. A total of 20 healthy finishing pigs (Duroc × Landrace × Large) were randomly divided into two groups. The control group (CON) was fed the basal diet, and the experimental group (BXG) was fed a basal diet with BXG instead of 10% corn for a period of 43 d. Compared to the CON group, the carcass weight, intramuscular fat content, and marbling score were significantly increased, while the drip loss, b^* value, and shear force of the BXG group were significantly reduced (p < 0.05). Gene expression analysis showed that the mRNA expression of lipid synthesis and oxidative-type fiber related genes was significantly increased in the BXG group (p < 0.05). Proteomic research revealed that the metabolic pathways of the BXG and CON groups differed significantly. A total of 36 differentially expressed proteins were identified, mainly related to energy metabolism, fatty acid degradation, and endocrine regulation pathways. However, the contents of glutamine, glutamate, proline, and other amino acids in the BXG group were significantly reduced (p < 0.05). Overall, this study has a positive effect on improving meat quality, but the specific mechanism needs to be further explored, which offers practical guidance for the application of BXG in producing higher-quality pork and further promotes its commercial application. Full article

Elevated carbon dioxide (eCO₂) levels can enhance crop yields but may simultaneously reduce quality, impacting both macronutrient and micronutrient concentrations, and potentially decreasing protein content in cereal grains. This study examined the effects of elevated CO₂ (eCO₂) and nitrogen (N) fertilization on crop growth, yield, and soil nitrogen cycling through a glass greenhouse experiment using Eutric Regosol soil. The experimental design incorporated two CO₂ gradients: ambient CO₂ (aCO₂) at approximately 410 ppm during the day and 460 ppm at night, and eCO₂ at approximately 550 ppm during the day and 610 ppm at night. Additionally, two nitrogen fertilization treatments were applied: no fertilizer (N0) and 100 mg N kg⁻¹ dry weight (DW) soil (N100). Crops were cultivated under two cropping systems: the monoculturing of fababean (Vicia faba L.) or wheat (Triticum aestivum. Yunmai) and the intercropping of both species. The results demonstrated that eCO₂ significantly enhanced the growth and yield of both fababean and wheat, particularly when nitrogen fertilization was applied. Nitrogen fertilizer application did not always enhance crop yield, considering the complexity of nitrogen management under elevated CO₂ conditions. Furthermore, the intercropping of fababean and wheat presented multiple advantages, including improved crop yields, enhanced soil health, and increased ecosystem services. These findings suggest that intercropping can serve as a sustainable strategy to boost productivity and ecosystem resilience in the face of climate change. The changes in nitrogen application and CO₂ concentration affect the gene copy number of ammonia-oxidizing bacteria and archaea, which may affect the nitrogen cycling process in soil. There are complex interactions between crop biomass, nitrogen accumulation, transpiration rate, photosynthetic rate and stomatal conductance with soil properties (e.g., pH, organic matter, nitrogen content) and microbial community structure. The interaction between CO₂ concentration, nitrogen application level and crop intercropping pattern had significant effects on crop growth, soil properties and microbial communities. Future research should prioritize investigating the long-term effects of intercropping on soil productivity and the development of management strategies that optimize the benefits of this cropping system. Full article

Familial hemiplegic migraine (FHM) is characterized by repeated episodes of reversible localized neurological deficits, in addition to headache. The aura of HM includes visual, sensory, motor, and verbal symptoms. Hemiplegic migraine (HM) is classified into non-familial sporadic HM (SHM) and familial HM (FHM). Here, we analyzed the clinical symptoms and their relevance in four Japanese patients considered to have SCN1A mutations as a cause. Sequencing of SCN1A was performed using a whole exome sequence method in 48 blood samples from clinically suspected patients with FHM. Subsequently, algorithm analysis, allele frequency determination, and three-dimensional structure analysis of the recognized variants were performed, and the recognized variants were evaluated. We found five heterozygous missense mutations (p.A23E, p.V250L, p.T398M, p.R1575C, p.L1660I) in SCN1A, three of which had not been reported. These five mutations may also affect the structure of the protein products, as assessed using a three-dimensional structural analysis. In all cases, the clinical symptoms included visual, sensory, motor, and verbal symptoms, which are forms of aura. Similarities were detected, such as the appearance of symptoms at a young age and other symptoms, such as hemiplegia after a headache attack. We report five missense mutations in SCN1A of Japanese cases. Full article

Understanding triacylglycerol (TAG) metabolism is crucial for developing algae as a source of biodiesel. TAGs are the main reservoir of energy in most eukaryotes. The final, rate-limiting step in the formation of TAGs is catalyzed by 1,2-diacylglycerol acyltransferases (DGATs). In the green alga Chlamydomonas reinhardtii, DGAT3 is phylogenetically related to plant DGAT3 but unrelated to other DGATs from eukaryotes, such as DGAT1 and DGAT2. In this study, we described the conformational preferences and the lipid-binding features of the DGAT3 from C. reinhardtii. To characterize its conformational stability and structural features, we used several biophysical probes, namely, fluorescence, circular dichroism (CD), and differential scanning calorimetry (DSC). Our results showed that the protein was mainly disordered, containing a small population of folded conformations in a narrow pH range (pH 8 to 10). The conformational stability of the folded structure of DGAT3 was very low, as shown by urea or guanidinium denaturations. Thermal denaturation, followed by fluorescence or CD, as well as calorimetric denaturation, followed by DSC, did not yield any transition in the pH range where DGAT3 acquired a “native-like” conformation. Furthermore, we used two approaches to demonstrate the interaction of DGAT3 with lipid membranes at the pH at which it had acquired a “native-like” conformation. The first involved the measurement of anisotropy and fluorescence quenching of the protein. The second approach focused on examining possible modifications of the biophysical properties of lipids due to their interaction with DGAT3, through anisotropy measurements and leakage assays. Both methods produced consistent results, suggesting that DGAT3 preferentially interacted with negatively charged membranes. These results will allow the design of a more efficient and stable DGAT3, as well as an in-depth understanding of how the metabolism of TAGs is accomplished in C. reinhardtii. Full article

Accurate quantification of indoxyl sulfate (IndS) and p-cresyl sulfate (pCS) is essential for understanding their role in chronic kidney disease (CKD) progression and for developing strategies to mitigate their harmful effects, including cardiovascular morbidity and renal fibrosis. Advances in liquid chromatography–high-resolution mass spectrometry (LC–HRMS) enable the integration of powerful diagnostic tools into clinical laboratories. Along with accurate quantification, precise mass measurements allow for untargeted compound identification. Methods. An LC–HRMS was validated for quantifying IndS and pCS in human serum, following EMA guidelines. The method involved protein precipitation with methanol, micro-LC for chromatographic separation, and detection based on accurate mass, with simultaneous high-resolution full-scan acquisition. Clinical samples from patients with varying degrees of renal insufficiency and samples obtained before and after hemodiafiltration were analyzed. Results. The method demonstrated acceptable linearity, precision, and accuracy. The measurement range for both analytes was from 100 to 40,000 ng/mL. Serum levels of IndS and pCS correlated with decreased renal function. After hemodiafiltration, there was a significant reduction of IndS (50%) and pCS (43%). Simultaneous untargeted analysis allowed to identify metabolites significantly underexpressed after hemodiafiltration. Conclusions. An accurate LC–HRMS method was validated for the quantification of IndS and pCS serum levels in patients with CKD, providing insights into toxin dynamics and enabling untargeted metabolic evaluation. Full article

Mutations in the TP53 gene and chromosomal instability (CIN) are two of the most common alterations in cancer. CIN, marked by changes in chromosome numbers and structure, drives tumor development, but is poorly tolerated in healthy cells, where developmental and tissue homeostasis mechanisms typically eliminate cells with chromosomal abnormalities. Mechanisms that allow cancer cells to acquire and adapt to CIN remain largely unknown. Tumor suppressor protein p53, often referred to as the “guardian of the genome”, plays a critical role in maintaining genomic stability. In cancer, CIN strongly correlates with TP53 mutations, and recent studies suggest that p53 prevents the propagation of cells with abnormal karyotypes arising from mitotic errors. Furthermore, p53 dysfunction is frequent in cells that underwent whole-genome doubling (WGD), a process that facilitates CIN onset, promotes aneuploidy tolerance, and is associated with poor patient prognosis across multiple cancer types. This review summarizes current insights into p53’s role in protecting cells from chromosome copy number alterations and discusses the implications of its dysfunction for the adaption and propagation of cancer cells. Full article

In most mammals, a withdrawal of the pro-gestational hormone progesterone (P4) is necessary for labor onset. In murine cervix, P4 withdrawal is mediated by enzymes steroid 5-alpha-reductase type 1 (SRD5A1) and 20-alpha-hydroxysteroid-dehydrogenase (20α-HSD). Previously, we have shown that inflammatory stimuli induce 20α-HSD levels in uterine muscle (myometrium). Here, we hypothesized that (1) infectious inflammation alters the levels of both P4-metabolizing enzymes in mouse cervix, which consequently ceases P4-mediated inhibition of cervical remodeling, thereby inducing preterm labor (PTL); (2) a progestin, selective progesterone receptor modulator promegestone (aka R5020), non-metabolizable by 20α-HSD, can block lipopolysaccharide (LPS)-induced PTL in mice by maintaining P4 signaling and preventing cervical remodeling. Using RT-PCR and IHC/IF methods, we evaluated the effect of inflammation on the expression of both enzymes in mouse cervix and determined if R5020 can prevent cervical remodeling and PTL in mice. We found significant induction of SRD5A1 and 20α-HSD proteins (p < 0.01), as well as transcript levels of pro-inflammatory cytokines Il1b, Il6, chemokines Cxcl1, Ccl2, cervical ripening enzyme Has2, hyaluronic acid binding protein/HABP (p < 0.05), and a simultaneous decrease in major extracellular fibrillar proteins, collagen type 1 and type 3 (col1a1, col3a1), in mouse cervix during PTL. The prophylactic administration of R5020 in pregnant mice significantly inhibited cervical remodeling and prevented PTL irrespective of the route of LPS-induction, systemic or local. We concluded that R5020 is a promising novel drug application for preterm birth prevention. Full article

Background: Products of lipid peroxidation include a number of reactive lipid aldehydes including reactive dicarbonyl electrophiles (DEs) and contribute to disease processes. DEs play a significant role in the development and progression of metabolic-associated steatotic liver disease (MASLD) by contributing to oxidative stress, inflammation, protein dysfunction, and mitochondrial impairment. Reducing DE stress may be a potential strategy for managing MASLD. We hypothesized that the DE scavenger 2-hydroxybenzylamine (2-HOBA) would reduce liver injury by reducing liver protein adduct formation by DE in mouse models of MASLD. Methods: Protein adducts were measured in human livers by immunohistochemistry and immunoblot. The effects of 2-HOBA were assessed in two different mouse models of MASLD. Results: Isolevuglandin (IsoLG) protein adducts were increased in MASH-staged human livers relative to histologically normal controls. Diet-Induced Animal Model of Nonalcoholic Fatty Liver Disease (DIAMOND) mice treated with 2-HOBA exhibited significantly lower fibrosis scores (* p = 0.012) and reduced liver transaminases (AST, p = 0.03) and ALT, p = 0.012) by over 40%. In STAM (Stelic Animal Model) mice, 2-HOBA improved NAFLD activity scores (p = 0.03, NAS), hyperglycemia, and inflammatory cytokines and reduced serum F2-isoprostanes (IsoPs) by 30%, p = 0.05. These improvements were absent mRNA changes in hepatic antioxidant enzymes (Cat, Gpx1, or Sod2) or ROS-generating proteins (p22PHOX, p47PHOX, NOX4 or COX1). Conclusions: DE scavenging with 2-HOBA may be a promising therapeutic strategy for managing MASLD. While findings are currently limited to male mice, a nutraceutical that reduces liver fibrosis could significantly improve the management of MASH by offering a non-invasive treatment option to potentially slow or reverse liver scarring, delay progression to cirrhosis, and improve patient outcomes, while also providing a potential treatment option for patients who may not be suitable for other interventions like liver transplantation. Full article

Background: Vaccines against COVID-19 target the spike protein. There is minimal information on longitudinal COVID-19 immune profiling in recovered versus naïve and vaccinated versus non-vaccinated healthcare workers (HCWs). Methods: This is a prospective longitudinal observational cohort of pediatric HCWs (pHCWs) conducted during 2020–2022 at an academic center, exploring the impact of COVID-19 vaccination on immunoglobulin G (IgG) antibody titers over time and cross-reactivity with other coronaviruses, including SARS-CoV-1, MERS-CoV, and seasonal coronaviruses (HCoV-HKU1 and HCoV-OC43). Results: A total of 642 pHCWs initially enrolled, and 337 participants had repeat IgG titers measured post-vaccine and post-booster. Most participants were female, median age range of 31–40 years. Anti-spike was higher in all vaccinated individuals versus non-vaccinated (p < 0.0001) and naïve versus infected (p < 0.0001). A single dose of vaccine was sufficient to attain maximum titers in recovered participants versus naïve who received both doses of vaccine. Anti-spike titers dropped significantly at 9 months after the primary series, whereas sustained anti-spike titers were observed at 9 months post-booster. Conclusions: All vaccinated pHCWs developed antibodies to spike. COVID-19 infection and/or vaccination yielded antibodies that cross-reacted to SARS-CoV-1, MERS-CoV, HCoV-HKU1, and HCoV-OC43. Anti-spike titers were more durable post-booster compared to the primary series. Longitudinal immune profiling of COVID-19 responses provides vital data to shape public health policies, optimize vaccine strategies, and strengthen pandemic preparedness. Full article