Search Results (4,508)

Background/Objectives: While COVID-19 severity is strongly associated with older age, younger adults may also develop complications. This study investigated factors associated with transfer to ICU in hospitalized younger adults during May–December 2021. Materials and methods: Eighty-six consecutive patients aged ≥18 and <50 years were admitted to the University Multi-profile Hospital for Active Treatment of Infectious and Parasitic Diseases “Prof. Ivan Kirov”, Sofia, Bulgaria between 12 May 2021, and 7 December 2021. Results: Among 86 patients (median age 40 years; 55.8% female), common symptoms included fever (89.5%), cough (91.9%), headache (76.7%), and gastrointestinal symptoms (83.7%). Laboratory abnormalities were frequent: lymphopenia (median 0.94 × 10⁹/L), elevated CRP (28.9 mg/L), LDH (333 U/L), and ferritin (198.5 µg/L). Severe andcritical disease occurred in 17.5% of cases, with 43% requiring oxygen therapy. In descriptive/univariate analyses, ICU transfer was more frequent among patients with hypoxemia (SaO₂ < 90%) and higher CRP, D-dimer, vomiting and ferritin. In a Firth penalized logistic regression model (rare-event setting), endocrine/metabolic comorbidity showed a directionally consistent association with ICU transfer; gastrointestinal comorbidity signals were not interpretable because of extremely small cell counts. Conclusions: Younger adults usually present with mild to moderate COVID-19; however, a substantial minority develop severe illness. Hypoxemia and elevated inflammatory and coagulation markers were consistently associated with worse outcomes. Endocrine/metabolic comorbidity showed the most consistent association with transfer to ICU in this cohort, although all comorbidity estimates should be interpreted cautiously, given the small number of events. Full article

Anthropogenic CO₂ emissions drive ocean acidification through changes in the carbonate system, lowering seawater pH. In contrast, salinity variations arise from physical processes such as freshwater fluxes and circulation. This study reports the preparation and Harned cell characterization of three equimolal TRIS buffer solutions (0.01 mol·kg⁻¹, 0.025 mol·kg⁻¹, and 0.04 mol·kg⁻¹) in artificial seawater (ASW) matrices with practical salinities of 35 and 50 and temperatures of 20 °C, 25 °C, and 30 °C. Determined pH_T values achieved expanded uncertainties ($U_{{\mathrm{p}\mathrm{H}}_{\mathrm{T}}}$ ≤ 0.006), meeting Global Ocean Acidification Observing Network (GOA-ON) “climate” quality standards. Absolute salinity (S_A) was concurrently measured via density (TEOS-10), revealing systematic deviations from practical salinity due to TRIS content. A nonlinear regression model was developed to predict pH_T as a function of salinity, temperature, and TRIS molality, with r² = 0.99998. These results provide a robust dataset for developing Certified Reference Materials (CRMs) for pH_T calibration under climate-relevant high-salinity environments at different temperature conditions, offering a practical tool for high-accuracy calibration in variable marine conditions. Full article

Pinus sylvestris essential oil (PSEO) has gained increasing interest as a natural anticancer candidate due to its bioactive phytochemical composition and potential to modulate apoptosis-related pathways. In this study, the chemical profile of PSEO was characterized by GC-MS, revealing oxygenated monoterpenes and monoterpene hydrocarbons as dominant constituents. Human brain (U-87MG) and peripheral nervous system (SH-SY5Y) tumor cells were treated with PSEO to evaluate cytotoxicity and mechanistic responses. Cell viability was assessed using the MTT assay, and 24-h IC₅₀ values were determined as 47.93 µg/100 µL for U-87MG and 71.63 µg/100 µL for SH-SY5Y, which were subsequently used for all mechanistic analyses. IC₅₀ exposure significantly increased intracellular ROS generation while reducing total antioxidant status, indicating oxidative stress-mediated cytotoxicity. Apoptosis-related ELISA assays demonstrated increased caspase-3 and caspase-9 activity, upregulated Bax, decreased Bcl-2, and a lowered Bcl-2/Bax ratio, collectively supporting the activation of the intrinsic mitochondrial apoptosis pathway. Molecular docking provided in silico evidence of favorable binding interactions between selected PSEO-associated ligands and apoptotic targets, consistent with experimentally observed biochemical outcomes. Overall, the findings suggest that PSEO exerts dose- and time-dependent anticancer effects and promotes mitochondrial apoptosis in U-87MG and SH-SY5Y cells, supporting its potential as a natural therapeutic candidate. Full article

PARP-1 and COX-2 have played important roles in several carcinomas, representing potential therapeutic targets; natural products have constituted interesting alternatives in cancer research, and complementary computational methods are relevant tools for the proposal of new molecules. Therefore, in this work, a theoretical study of a set of four derivatives of perezone and isoperezone, i.e., α-pipitzol, β-pipitzol, α-isopipitzol, and β-isopipitzol, employing quantum chemistry, bioinformatics, and docking, was performed. Conformational studies were accomplished to obtain minimum energy structures. Subsequently, they were optimized by the B3LYP hybrid method and the 6-311++G(d,p) basis set. With this same level of theory, the geometrical, electronic, and spectroscopic properties and the reactivity parameters were determined; moreover, a molecular docking evaluation was performed to determine their activity towards COX-2 and PARP-1. Additionally, a cytotoxicity activity assay was performed against various cancer cell lines; thus, α-pipitzol and β-pipitzol showed the greatest affinity for COX-2, and the α-isopipitzol exhibited two relevant interactions. Regarding α-pipitzol, it exhibited both affinity and an important interaction with PARP-1. Regarding β-pipitzol, it displayed the lowest inhibitory concentration in A549 (64.49 µM); nevertheless, α-isopipitzol presented the lowest inhibitory concentrations, 83.59 µM and 87.85 µM for U37 and MCF-7 cell lines, respectively. Full article

Collagenases can specifically degrade collagen, showing a wide application prospect in food, leather, waste utilization, biotechnology, and other industries. Currently, Hathewaya histolytica is commonly used in industry to produce collagenases, but its application is greatly limited by its pathogenicity. This study first identified five potential Pseudomonas-derived collagenases by sequence alignment. Bioinformatics tools were used to analyze their structures and functions. Heterologous expression of two P. chlororaphis-derived collagenases was achieved in E. coli, and their enzymatic properties were characterized. Bioinformatics analysis shows that the Pseudomonas-derived collagenases had low molecular weights (22.1~50.5 kDa) and good thermal stability (aliphatic index 73.73~88.81). Deletion of P. chlororaphis GP72ANO strain colP1 and colP2 genes had no significant effect on cell growth. The yields of collagenase ColP1 and ColP2 obtained from E. coli BL21(DE3) cultivation broth were 148 mg/L and 322 mg/L, respectively. The optimum temperature of each collagenase was 28 °C, and the soluble collagen activities of ColP1 and ColP2 were up to 42.64 U/mg and 21.21 U/mg, respectively. Collagenase ColP1 had the highest enzyme activity at pH 8, while collagenase ColP2 had the highest enzyme activity at pH 4. Metal ions such as Na⁺, K⁺, Mg²⁺, Ca²⁺, Ni²⁺, and Mn²⁺ inhibited the activity of collagenases to different degrees. This study successfully achieved recombinant expression and preliminary purification of Pseudomonas-derived collagenases in E. coli and explored their function and physicochemical properties. Full article

Background: Ideal plant architecture is central to high-yield maize breeding. The proportional length of internodes above the ear plays a crucial role in determining plant architecture. Methods: In this study, we used an association panel comprising 288 maize inbred lines and performed genome-wide association studies (GWASs) with 1.25 M high-density single nucleotide polymorphism (SNP) markers under a Q + K mixed linear model. Results: A total of 821 significant SNPs associated with plant height (PH), height above ear (HAE), and internode-related traits were detected, which were further classified into 417 quantitative trait loci (QTL). Among these, 128 significant SNPs and 44 QTL were identified for the U1/HAE trait, and 37 significant SNPs and 27 QTL for the U1/PH trait. Four stable QTL (qU1–qU4) were identified through colocalization analysis. Two candidate genes, Zm00001d013222 (involved in gibberellin signaling) and Zm00001d021304 (involved in cell wall metabolism), were further supported by haplotype analysis. For the former gene, U1/PH values in Hap1 and Hap3 were significantly lower than those in Hap2 (p < 0.01). For the latter gene, Hap2 exhibited a significantly higher U1/HAE value compared to Hap1 (p < 0.001). Conclusions: These findings provide new genetic insights into the regulation of maize internode proportion and plant architecture, offering potential targets for molecular breeding. Full article

Background: The tumor microenvironment (TME), composed of diverse immune and stromal cells, plays a key role in cancer progression. Among its components, tumor-associated neutrophils (TANs) and the desmoplastic reaction (DR) have emerged as important modulators of tumor behavior. While each has been extensively studied, their interrelationship and association with tumor grade and clinicopathological parameters remain unclear. Aim: This hypothesis-generating study aimed to explore the relationship between the presence of TANs, various types of DR, the grade of tumor malignancy, and other fundamental clinicopathological characteristics commonly studied in daily clinical practice. Materials and Methods: The study included a cohort of 65 cancer patients (N = 65). The average number of TANs was recorded. In hematoxylin and eosin (H&E)-stained sections, “hot spots” representing areas with the highest neutrophil density were first identified. The tumor-associated polymorphonuclear neutrophils were then counted in ten consecutive high-power fields (HPFs). In the same specimens, the DR was assessed and classified according to stromal texture. Results: TANs did not follow a normal distribution across any clinicopathological category (p < 0.05). Significant differences in TAN levels were observed among DR types (Kruskal–Wallis H = 9.890, p = 0.007), with higher counts in myxoid compared to mature stroma (Mean Rank = 41.58 vs. 24.80, p = 0.006). TAN levels also varied significantly with tumor grade (H = 22.384, p < 0.001), increasing from Grade 1 to Grade 3 (p < 0.013–0.001). Higher TAN counts were associated with cellular erythroblastic oncogene B2 (c-erbB2) positivity (H = 6.547, p = 0.038), perineural invasion (Mann–Whitney U = 179.5, p < 0.001), and ER/PR negativity (p = 0.016 and p = 0.044, respectively). No significant association was found with necrosis (p = 0.083). A near-significant relationship was identified between DR type and tumor differentiation grade (χ² = 9.448, p = 0.051), with mature stroma most common in Grade 1 tumors, keloid-like stroma in Grade 2, and myxoid stroma in Grade 3. Conclusions: High TAN levels were linked to aggressive tumor features and specific DR patterns. The association of myxoid stroma with elevated TAN infiltration may reflect a highly aggressive TME. These preliminary results warrant validation in larger, prospective studies. Full article

Background/Objectives: Although 90% of prostate cancer (PCa) metastasis occurs in the bone, there are limited studies and rarely available genome-wide profiles at individual sample level for genomic copy number changes in the literature. Methods: We performed Affymetrix SNP 6.0 high-density microarray analysis to generate the genome-wide copy number change profiles for six cases of PCa bone metastases. A common genomic loss was confirmed by fluorescence in situ hybridization (FISH) in paraffin-embedded PCa bone metastasis samples together with primary PCa and benign prostate hyperplasia samples. We overexpressed the candidate miRNA in PCa cell lines and knocked down its target genes by siRNA transfection and investigated the effect on protein expression and cell viability, migration, and invasion abilities, respectively. Protein expression in PCa tissues was analyzed by immunohistochemical staining. Results: We provided high-resolution PCa bone metastasis profiles of six cases and identified potential bone metastasis-specific common genomic alterations, including a 1.6 mb region on 17q11.2, as well as those shared by non-bone metastatic PCa. The common 17q11.2 loss was confirmed by FISH in further 14/21 PCa bone metastasis samples but was only found in 9/151 primary PCa samples. The well-established tumor-suppressing miRNA located within this small genomic region, miR-193a-3p, was downregulated in both bone metastasis and primary PCa cases, leading to overexpression of cyclin D1 and uPA to promote cancer cell migration and invasion. Cyclin D1 was highly expressed in both localized PCa and bone metastasis samples, and the expression was significantly higher in the latter group (p = 0.013). Conclusions: We generated high-resolution copy number change profiles for bone metastasis samples. This led to the identification of a common, small genomic loss and downregulation of miR-193a-3p, which suppresses PCa bone metastasis through inhibition of its target proteins, providing new insight into bone metastasis development. Full article

The escalating global incidence of ulcerative colitis (UC) underscores the demand for novel therapeutic strategies. This study investigated the fermentation of Periplaneta americana (PA) powder using two conventional dairy starter strains, Lactobacillus delbrueckii subsp. bulgaricus SN22 and Streptococcus thermophilus SN05, to enhance its functional properties, particularly anti-inflammatory activity, via microbial processing. Both strains demonstrated favourable safety and antimicrobial activity. Untargeted metabolomics revealed that fermentation significantly altered the metabolite profile of the PA supernatant, enriching compounds with potential bioactivities, notably anti-inflammatory (e.g., 3-anisic acid) and antioxidant (e.g., vitamin U) properties. In the DSS-induced mouse colitis model, treatment with the fermented supernatant alleviated intestinal inflammation compared to the unfermented group. This was demonstrated by significantly reduced levels of the pro-inflammatory cytokines IL-1β and TNF-α, along with improved maintenance of intestinal barrier integrity. Further in vitro assays showed that the fermented supernatant significantly suppressed proliferation and clonogenicity in human HT-29 colon cancer cells, while also inducing reactive oxygen species accumulation and apoptosis. Results demonstrate these strains are multifunctional starters possessing superior antimicrobial and anti-inflammatory efficacy. This study employed LAB fermentation of insect-derived matrices to derive bioactive components. The fermentation products exhibited anti-inflammatory potential, offering a potential microbial transformation strategy for developing functional products for adjunctive UC intervention. Full article

HIV-1 entry into host cells culminates in integration of the reverse transcribed double-stranded viral DNA into host genes. Several lines of evidence suggest that intact, or nearly intact, HIV-1 cores—large, ~60 nm-wide structures—pass through the nuclear pore complex (NPC), and that this passage is associated with pore remodeling. Cryo-electron tomography studies support the dynamic nature of NPCs and their regulation by cytoskeleton and ATP-dependent processes. To explore NPC remodeling, we used super-resolution Stochastic Optical Reconstruction Microscopy (STORM) of U2OS cells endogenously expressing nucleoporin 96 tagged with SNAP. Single-molecule localization imaging and computational averaging resolved 8-fold symmetric nuclear pores with an average radius of ~51 nm. Depletion of cellular ATP using sodium azide or antimycin A, previously reported to reduce the size of yeast NPCs, did not significantly alter the nuclear pore radius in U2OS cells. Similarly, stressing the nuclear envelope by hypotonic or hypertonic conditions failed to induce detectable expansion or contraction of NPCs. These results indicate that the NPCs in U2OS cells do not respond to ATP depletion nor mechanical stresses on changes in pore morphology that can be resolved by STORM. Since these cells are infectable by HIV-1, we surmise that direct multivalent interactions between HIV-1 capsid and phenylalanine-glycine nucleoporins lining the pore’s interior drive the core penetration into the nucleus and the associated changes in the pore structure. Full article

Background/Objectives: The development of small-molecule agents that selectively target DNA replication remains a central strategy in anticancer drug discovery. In this study, we report the biological characterization of a novel 6-nitro-benzodioxin-naphthalimide (NI) derivative (compound 5a), evaluated as a potential DNA-targeted anticancer lead. Methods/Results: The antiproliferative activity of 5a was assessed in a small panel of human lung carcinoma cell models (A549, H1299) and a non-malignant control (MRC-5), revealing pronounced cytotoxic effects in tumor cells, accompanied by favorable selectivity indices. Mechanistic investigations demonstrated that treatment with 5a results in strong inhibition of DNA synthesis, as evidenced by a marked reduction in EdU incorporation and a robust induction of the DNA damage marker γH2AX. These effects were associated with cell-cycle perturbations characterized by accumulation in G₁ and G₂/M phases, followed by activation of apoptotic pathways. Importantly, clonogenic survival assays confirmed that even transient exposure to 5a leads to a sustained loss of proliferative capacity, indicating irreversible long-term cellular damage. These results support a replication stress-driven mechanism of action for compound 5a, consistent with interference in DNA-associated processes during S phase. Conclusions: While the precise molecular initiating event remains to be elucidated, the observed biological profile positions 5a as a promising DNA-targeted lead structure with potential for further pharmaceutical optimization. These findings provide a solid foundation for the continued development of naphthalimide-based compounds as anticancer agents within a pharmaceutically relevant framework. Full article

Recurrent pregnancy loss (RPL) is defined as the loss of two or more pregnancies before the 22nd gestational week and affects 10–15% of clinical pregnancies. Despite extensive diagnostics, over 50% of RPL cases remain unexplained, suggesting an important role for immunological mechanisms. Sex hormones (SH) are key regulators of immune responses during pregnancy; however, their influence on immune checkpoint proteins (ICPs) is poorly understood. This study evaluated the effects of progesterone, β-estradiol, and dihydrotestosterone (DHT) on ICP expression on immune cells, including Treg, NK, NKT, TC, Th, and T cells, collected from pregnant women and patients with unexplained RPL (uRPL). Peripheral blood mononuclear cells from 20 pregnant women and 20 uRPL patients were cultured for 48 h with SH. The expression of the first generation of ICPs—PD-1 and TIM-3—and the second—LAG-3, TIGIT, and VISTA—on T, NK, and NKT cells was analyzed by the flow cytometry method. In pregnant women, SH exerted modest effects, with DHT increasing VISTA and LAG-3 expression, while progesterone and estradiol mainly upregulated LAG-3 and TIM-3 on cytotoxic cells. In contrast, uRPL immune cells showed pronounced SH sensitivity, characterized by increased TIM-3 and VISTA expression and reduced TIGIT expression, particularly after DHT stimulation. In conclusion, SH modulates ICP expression in a cell-specific manner, with stronger effects observed in uRPL patients’ lymphocytes. These findings highlight a potential role for hormonal and ICP-targeted strategies in RPL management. Full article

Citrus reticulata Blanco peel is a dominant industrial waste that was recently revalued as a source of bioactive molecules. This study explored its phytochemical and antileukemic potentials. The bioassay-guided fractionation of the dichloromethane extract yielded nobiletin, scoparone, and tangeretin, which were identified spectroscopically. The extract, fractions, and compounds showed antiproliferative effects in both THP-1 and U937 cells, which were employed as in vitro models of acute myeloid leukemia (AML). According to cytofluorimetric analysis, the extract and fractions inhibited cell growth by both apoptosis and necrosis, whereas the single molecules induced apoptosis. This mechanism was mediated by the modulation of BAX and BCL-2 genes in both AML cell lines. Notably, each treatment drove THP-1 and U937 cells into the sub-G0 phase, together with an increase in the cell population in the G1 phase of the cell cycle, both of which were detected cytofluorimetrically. In line with these findings, the extract, fractions, and single molecules counteracted the overexpression of CYCLIN A1 in THP-1 cells while reducing the expression of CYCLIN D2 in U937 cells. Moreover, cell treatments attenuated the invasiveness of AML cells through the upregulation of TIMP-2 at the transcriptional level. Therefore, this study supports pharmaceutical interest in citrus waste for cancer management, providing evidence on its antileukemic potential in vitro. Full article

Fengycin, a lipopeptide synthesized by Bacillus species, exhibits pronounced antifungal activity; however, its low production titer remains a primary constraint to broader application. Global regulatory factors constitute key actionable targets for enhancing microbial synthesis. Here, we verified the ability of Bacillus amyloliquefaciens TF28 to produce fengycin with potent inhibitory activity. Transcriptomic analysis identified five global regulators linked to fengycin biosynthesis in this strain. Following their overexpression, fermentation kinetics indicated that while these regulators generally did not affect glucose utilization, each exerted a distinct effect on cell growth and fengycin production. Specifically, degQ overexpression increased fengycin production to 116.0 mg/L, corresponding to a 23.40% increase relative to the strain without degQ overexpression, whereas overexpression of degU, sigmaH, phoP, and abrB reduced it. Moreover, degQ and abrB overexpression modulated the expression of key fengycin synthetase genes, including fenA and fenB. Collectively, these findings establish that degQ, degU, sigmaH, phoP, and abrB functionally regulate fengycin biosynthesis in B. amyloliquefaciens TF28, providing a conceptual framework for the rational design of engineered strains with enhanced fengycin productivity. Full article

Objectives: Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is an autoimmune encephalitis that can lead to severe neurological impairments, particularly in pediatric patients. Effective biomarkers for diagnosis and prognosis are crucial for improved treatment outcomes. To evaluate the potential of soluble Triggering Receptor Expressed on Myeloid cells 2 (sTREM2) in cerebrospinal fluid (CSF) and serum as diagnostic and prognostic biomarkers in pediatric patients with anti-NMDAR encephalitis. Methods: The study included 21 children diagnosed with anti-NMDAR encephalitis and 27 children with non-inflammatory neurological disorders (OND) as controls. CSF and serum samples were collected from each patient. sTREM2 levels were measured using enzyme-linked immunosorbent assay (ELISA). Statistical analyses, including Mann–Whitney U test and ROC curve analysis, were performed to assess the diagnostic and prognostic value of sTREM2. Results: sTREM2 levels in CSF and serum were significantly higher in children with anti-NMDAR encephalitis compared to the OND group (p < 0.001). CSF sTREM2 levels showed a positive correlation with modified Rankin Scale (mRS) scores and a negative correlation with Glasgow Coma Scale (GCS) scores, suggesting an association with disease severity. ROC curve analysis demonstrated that CSF sTREM2 had a high diagnostic accuracy (AUC = 0.887, p < 0.001), while serum sTREM2 showed a slightly lower diagnostic accuracy (AUC = 0.848, p < 0.001). Patients with better prognoses had significantly lower CSF sTREM2 levels than those with poorer outcomes (p = 0.029). Conclusions: Elevated CSF sTREM2 levels were associated with increased neuroinflammation and poorer clinical outcomes in children with anti-NMDAR encephalitis. These findings suggest that CSF sTREM2 may serve as a valuable biomarker for the diagnosis and prognosis of pediatric anti-NMDAR encephalitis. Full article