Search Results (267)

Oncolytic coxsackievirus B3 (oCVB3) strain PD-H has shown potent oncolytic efficacy and a remarkable safety profile in the treatment of colorectal cancer in vivo after intratumoral (i.t.) injection. In this study, we investigated the safety and efficiency of PD-H following intravenous (i.v.) virus administration. When injected i.v. into Balb/C mice bearing subcutaneous Colon-26 tumors, PD-H led to slightly reduced tumor progression and a significant increase in animal survival, but it also caused multi-organ infection and tissue damage. To improve the safety profile of PD-H, we inserted microRNA target sites (miR-TS) of the heart-specific miR-1, pancreas-specific miR-375, liver-specific miR-122, and brain-specific miR-124 or the tumor-suppressor miR-145 into the genome of PD-H and generated the viruses PD-622TS and PD-145TS. Both viruses replicated similarly and induced cytotoxicity comparable to that of PD-H in the colorectal carcinoma cell lines Colon-26 and CT-26Luc. Their replication was inhibited in HEK293T cells transiently transfected with the cognate microRNAs. In vivo, i.v. administration of PD-145TS and PD-622TS to healthy Balb/C mouse resulted in significantly lower viral titers in the organs of mice and led to significantly less-intense pathological alterations compared to PD-H. PD-622TS injected i.v. into Balb/C mice with CT-26Luc-induced peritoneal carcinomatosis did not induce off-target alterations in normal organs, but it failed to induce a therapeutic effect. These data indicate that PD-H or microRNA-regulated PD derivatives exhibit only limited therapeutic efficacy following i.v. injection in colorectal tumor-bearing mice. However, the newly engineered microRNA-regulated PD-H variants demonstrate improved safety profiles. Full article

The accumulation of proline, an important osmoprotective and antioxidant compound, is a key defense mechanism induced in plants in response to stress factors, including salinity, and is likely dependent on abscisic acid (ABA). However, in barley grown for 8 days under salinity conditions (125 mM NaCl), proline accumulation was not accompanied by changes in ABA content. Co-application of 0.5 mM molybdenum (Mo) significantly reduced NaCl-induced oxidative stress, as measured by H₂O₂, O₂⁻, MDA, and chlorophyll content, and increased the activity of Mo-containing aldehyde oxidase (AO), an enzyme involved in de novo ABA synthesis. As a result, elevated ABA levels were observed, but proline content under salinity conditions was similar in Mo-treated and non-Mo-treated plants. In contrast, exposing plants to 0.5 mM tungsten (W), an antagonist of Mo, inhibited AO activity without significantly altering ABA content, while proline and oxidative stress marker levels increased dramatically under both non-saline and saline conditions. The observed changes in proline content are mainly due to modulation of the rate of synthesis and, to a lesser extent, the rate of degradation, as revealed by transcript abundance of P5CS1 and PDH, which encode D1-pyrroline-5-carboxylate synthetase and proline dehydrogenase, respectively. The results indicate that in barley grown under salinity conditions, proline accumulation is ABA-independent but depends on the level of oxidative stress modulated by Mo and W ions. Full article

This study investigated the mechanism by which combined vacuum packaging and 0.5% (w/v) ascorbic acid treatment (VP-AsA) preserves fresh-cut potatoes at 4 °C, integrating physiological and transcriptomic analyses. Transcriptome sequencing revealed 2246 differentially expressed genes (DEGs) in the VP-AsA group. Notably, key genes involved in glutathione metabolism and NADPH regeneration—encoding glutathione reductase (GR), glutathione S-transferase (GST), isocitrate dehydrogenase (IDH), glucose-6-phosphate dehydrogenase (G6PDH), and ornithine decarboxylase (ODC)—were significantly up-regulated. This transcriptional reprogramming, which was associated with increased glutathione (GSH) content, provides a molecular basis for the enhanced antioxidant capacity observed in the treated samples, including elevated superoxide dismutase (SOD) activity, DPPH/ABTS radical scavenging capacity, and ferric reducing antioxidant power (FRAP). Concurrently, VP-AsA treatment reduced water migration, inhibited polyphenol oxidase (PPO) and peroxidase (POD) activities, and maintained key textural properties (hardness, fracturability, springiness, chewiness) during the first 9 days of storage. These results suggest that VP-AsA treatment preserves quality at least in part by transcriptionally activating glutathione-mediated antioxidant pathways, providing insights for fresh-cut fruits and vegetables quality control. Full article

Background/Objective: Oblique lumbar interbody fusion (OLIF) achieves indirect decompression through restoration of disc height. Because maintenance of the restored disc space is essential for sustained neural decompression, solid fusion without cage subsidence is a key determinant of successful surgical outcomes. This study aimed to evaluate preoperative and intraoperative predictors of cage subsidence and radiographic fusion after OLIF. Methods: Seventy patients (119 levels) who underwent OLIF using a polyether–ether–ketone cage and posterior screw fixation between 2015 and 2023 were retrospectively reviewed. Preoperative bone quality was assessed using the computed tomography-based Hounsfield unit (HU) and magnetic resonance imaging-based vertebral bone quality (VBQ) score on T1-weighted images. Radiographic parameters of anterior and posterior disc height (ADH, PDH), segmental and lumbar lordotic angle (SLA, LLA), foraminal height (FH), and cage position were measured preoperatively at one-year follow-up. Results: Cage subsidence occurred in 21.0% of spinal levels (25/119 levels). Multivariate analysis identified these measures as independent predictors: HU (OR 1.017; p = 0.012), VBQ score (OR 2.716; p = 0.016), and PDH distraction (OR 1.418; p = 0.019). ROC analysis identified cutoff values of HU < 145.86 (AUC = 0.654), VBQ score > 3.30 (AUC = 0.723), and PDH distraction > 4.79 mm (AUC = 0.672). None of the evaluated factors were significantly associated with one-year radiographic fusion. Conclusions: Lower HU, higher VBQ score, and excessive PDH distraction are independent risk factors for cage subsidence after OLIF, although these factors do not appear to affect short-term fusion outcomes. Full article

The ketogenic diet (KD) is a metabolic intervention characterized by high fat and very low carbohydrate intake, showing significant metabolic, neuroprotective, and therapeutic effects. However, its efficacy varies widely due to individual genetic and epigenetic factors. This review synthesizes current knowledge of genes most strongly associated with KD response, including polymorphisms in FTO, APOA2, PPAR, SCN1A, KCNQ2, STXBP1, CDKL5, the MODY gene group, and SLC2A1, which shape outcomes across lipid metabolism, energy expenditure, inflammation, and neurotransmission. Epigenomic modifications induced by a KD, such as changes in DNA methylation and histone acetylation involving BDNF, SLC12A5, KLF14, and others, modulate functional metabolic and neurological effects. Sex and age further modulate KD effects through distinct patterns of gene activation and hormonal interactions. These variables together impact metabolic and neurological outcomes and are critical for developing personalized nutrition and disease management strategies. Based on the reviewed evidence, genetic and epigenetic profiling can help identify patients who are likely to benefit from a KD (e.g., GLUT1DS, PDH deficiency) and those in whom a KD may be ineffective or harmful (e.g., SCOT or SLC2A1-independent defects). The review concludes that genetic and epigenetic profiling is recommended for personalized dietary interventions. Full article

Low-temperature stress is a major factor that limits the productivity and geographical distribution of melon (Cucumis melo L.). This study elucidates that CmDUF239-1 is a positive regulator of cold stress, and its underlying mechanisms are investigated using root-specific overexpression lines. Seedlings overexpressing CmDUF239-1 exhibited increased biomass and reduced relative electrical conductivity under cold stress. CmDUF239-1 overexpression promoted the accumulation of soluble sugar and proline, which was accompanied by enhanced activity of the proline biosynthetic enzyme Δ¹-pyrroline-5-carboxylate synthase (P5CS) and suppressed activity of the proline-degrading enzyme proline dehydrogenase (PDH). Molecular analysis revealed that CmDUF239-1 overexpression upregulated antioxidant enzyme-related genes, sugar metabolism related genes, and proline-related genes. Furthermore, it activated key genes in the ICE-CBF-COR signaling pathway, including CmCBF1, CmCBF2, and the downstream effector gene CmCOR413-2. In conclusion, the CmDUF239-1 gene enhances melon cold tolerance by modulating antioxidant defense, enhancing osmolyte (sugar and proline) metabolism and activating a core signaling pathway. This study not only characterizes a novel function for a DUF family gene but also provides a promising candidate gene for the genetic improvement of cold resilience in melon and other related crops. Full article

This study addressed the optimal magnesium (Mg) requirement for juvenile largemouth bass (Micropterus salmoides) and assessed the effects of dietary Mg supplementation on growth performance, nutrient metabolism, and alleviation of heat stress in it. In this study, six diets with varying Mg levels (1.01, 1.26, 1.78, 2.24, 2.35, and 2.51 g/kg), designated as MG1, MG2, MG3, MG4, MG5, and MG6, respectively, were formulated using MgSO₄·7H₂O as the Mg source. These diets were fed to juvenile M. salmoides (initial body weight 2.27 ± 0.02 g) for 8 weeks. The growth performance of the MG4 group was significantly improved. In addition, Plasma GLU, LDL-C, and TG levels were significantly reduced in the MG4 group, while plasma HDL-C levels were increased. In terms of gene expression, glut2, g6pdh, ppar-γ, fas, elovl2, acc, and igf-1 were significantly upregulated in the MG4 and MG5 groups, while g6pase and ppar-α were significantly downregulated in the MG5 group. In the heat stress test, MG4 group exhibited enhanced antioxidant capacity, as evidenced by decreased plasma MDA levels and increased CAT activity, coupled with enhanced gill Na⁺/K⁺-ATPase activity. Gene expression results also showed that il-10 and bcl-2 were significantly upregulated in the MG4 group, while nf-κb, ifn-γ, il-8, tnf-α, casp3, casp8, bax, jnk2 and ask1 were significantly downregulated. Furthermore, the results of TUNEL immunofluorescence labeling analysis showed that the apoptotic index was significantly decreased in the MG2-MG6 groups. Overall, appropriate dietary Mg levels promoted growth performance, improved glucose metabolism, and induced lipid deposition in juvenile M. salmoides. Notably, Mg reduced oxidative damage by enhancing antioxidant enzyme activity, thereby modulating heat stress-induced Antioxidant–Inflammatory–Apoptotic of juvenile M. salmoides. Based on quadratic regression analysis of SGR and FCR, the optimal Mg requirement for juvenile M. salmoides was 2.04, and 2.15 g/kg, respectively. Full article

Background: Community-onset fungemia is a clinically significant syndrome frequently linked to recent healthcare exposure and significant morbidity and mortality. Methods: We performed a 21-year, single-centre retrospective cohort of consecutive yeast bloodstream infections diagnosed at the Emergency Department (2004–2024). Clinical/epidemiological data, species identification (MALDI-TOF MS), antifungal susceptibility (CLSI M27; Sensititre YO10), and whole-genome sequencing (WGS) were analyzed. Results: Forty-eight episodes (51 isolates) were included; 56.3% were male, median age 74 years (IQR 63–82). Acquisition was healthcare-associated in 38/48 (79.2%). Sources were unknown (36.7%), abdominal (22.4%), urological (22.4%), catheter-related (14.3%), and 2.1% was attributed to a cardiovascular and a joint focus; 18.8% were polymicrobial. Crude mortality was 20.8% at 7 days (10/48) and 29.2% at 30 days (14/48). Species distribution: Candida albicans 41.2%, Nakaseomyces glabratus 27.5%, Candida parapsilosis 11.8%, Candida tropicalis 11.8%, Pichia kudriavzevii 3.9%, Clavispora lusitaniae 1.9%, and Candida orthopsilosis 1.9%. No isolate was resistant to anidulafungin, micafungin, or amphotericin B; one N. glabratus showed reduced susceptibility to caspofungin. Azole resistance was observed in one C. albicans and one N. glabratus isolate. WGS (44 isolates) confirmed MALDI-TOF identifications and characterized resistance markers. All 12 sequenced N. glabratus carried ERG2 I207V, PDR15/PDH1 E839D, and PDR1 V91I/L98S. Notable cases included one N. glabratus caspofungin-intermediate with FKS2 F659C, N. glabratus fluconazole-resistant with multiple PDR1 substitutions including a unique novel G857V, and C. albicans fluconazole-resistant harbouring alterations in MRR1/MRR2, CDR1, and ERG11. Conclusions: In this 21-year cohort, community-onset fungemia was predominantly healthcare-associated, with C. albicans as the predominant species, followed by N. glabratus. Crude mortality reached 29.2% at 30 days. Echinocandin resistance was not observed; azole resistance was uncommon. WGS provided precise speciation and actionable insight into resistance mechanisms, including a putatively novel PDR1 G857V in N. glabratus. Full article

Introduction: Currently, static interbody cages are the gold standard for achieving solid arthrodesis in the spine, enhancing segmental stability, obtaining neuroforaminal decompression, and improving as well as maintaining segmental lordosis. It is well known that restoring sagittal balance and segmental lordosis is crucial for long-term outcomes in lumbar spine fusion. For some cases, expandable interbody cages are emerging as an alternative to static cages. This study aims to evaluate the radiographic outcomes and complications of standard open transforaminal lumbar interbody fusion (TLIF). Methods: A standard open TLIF procedure using expandable cages was performed at 1 to 3 levels in 71 patients (129 levels in total), with a follow-up of two years. All patients underwent radiological assessments preoperatively, immediately postoperatively, and at one and two years postoperatively. Radiological evaluation was conducted using standing lateral X-rays. Results: Segmental lordosis (SL) increased significantly from the preoperative value (9.0° ± 3.6°) to 24 months postoperatively (15.4° ± 3.0°), with improvements maintained throughout the 24-month follow-up period (p < 0.001). Similarly, anterior disc height (ADH), posterior disc height (PDH), and foraminal height (FH) each increased significantly from preoperative to immediate postoperative measurements, and these gains were maintained over the two-year follow-up (p < 0.001 each). Lumbar lordosis increased significantly from the preoperative value (41.9° ± 10.5°) to the immediate postoperative period (45.7° ± 10.8°); however, this improvement decreased slightly at the one- and two-year follow-ups. No revisions were required for cage-related complications. One patient experienced a surgical site infection, and two patients had mechanical complications (screw loosening and proximal junctional kyphosis). Conclusions: Expandable interbody cages enable excellent restoration and maintenance of disc height and segmental lordosis in a standard open TLIF procedures at two-year. Achieving these outcomes depends on several factors, including proper preparation of the vertebral endplates, accurate cage placement and expansion, posterior facet osteotomy, and the application of posterior compression prior to final fixation. These steps are essential to fully maximize the potential of expandable cage technology. Full article

Previous studies have demonstrated that melatonin (MLT) enhances boar sperm motility by modulating energy metabolism status, yet the underlying mechanisms remain incompletely understood. This study aims to investigate whether sirtuin 3 (SIRT3), a key mitochondrial deacetylase, mediates MLT’s effects. Herein, the semen of six Landrace boars (16–18 months of age) was treated with 1.0 μM MLT with/without the SIRT3 inhibitor 3-TYP, preserved at 17 °C for 3 days, and subsequently maintained at 37 °C for a duration of 10 min. We demonstrated that MLT upregulated SIRT3 protein expression and reduced the acetylation level in mitochondrial proteins. MLT significantly increased glucose uptake and suppressed lactate release in the sperm, while elevating levels of pyruvate and acetyl-CoA, the substrates of pyruvate dehydrogenase (PDH) and the tricarboxylic acid (TCA) cycle, respectively, and the protein expression of PDH, indicating enhanced metabolic flux. Notably, inhibition of SIRT3 reversed MLT’s effects: it blocked the increases in SIRT3 expression, glucose consumption, PDH expression, complex I activity, ATP content, and superoxide dismutase (SOD) activity, and prevented the decreases in the levels of acetylation and lactate, as well as pyruvate kinase (PK) activity, confirming the essential role of SIRT3. Functionally, the MLT-induced improvements in sperm motility parameters (total, progressive, fast motility, immotile) were also reversed by 3-TYP. Collectively, these findings demonstrate that the SIRT3-mediated pathway is essential for MLT to enhance boar sperm energy metabolism and antioxidant defense, thereby increasing ATP production and enhancing sperm motility. Targeting SIRT3 represents a promising therapeutic strategy for improving boar fertility and may also provide insights for research into human male infertility. Full article

Chinese cordyceps, a highly valued traditional medicine, is formed when the fungal parasite Ophiocordyceps sinensis infects an underground caterpillar (Thitarodes). This interaction progresses slowly, as the larvae have a long developmental period and the fungus requires several months to complete its growth. The medicinal fungus O. sinensis has a complex life cycle that requires successful transformation from blastospores to hyphae for the formation of Chinese cordyceps. Building on our previous identification of diverse microbial communities associated with Thitarodes xiaojinensis larvae, this study investigates the role of host microbiota in enhancing O. sinensis blastospore transformation under in vitro conditions. Cultured supernatant of Fusarium pseudonygamai associated with T. xiaojinensis larvae significantly increased transformation rates by 31.6% after 8 days. Transcriptomic profiling revealed early upregulation of genes involved in energy metabolism, stress response, detoxification, and hyphal morphogenesis (notably CYC1, hmp, gedE, and fahA), supporting the cellular reprogramming required for fungal development. Additionally, mannitol isolated from F. pseudonygamai culture acted as a key promoter of transformation. Further functional assays confirmed that modulation of mannitol biosynthesis genes (M1PDH and MDH) through chemical agonists and inhibitors directly influenced mannitol levels and transformation efficiency. Collectively, these results highlight the pivotal role of microbiota-derived metabolites, particularly mannitol, in regulating O. sinensis transformation, offering potential strategies to improve artificial cultivation of Chinese cordyceps. Full article

Hypocrellin A (HA), a photodynamic perylenequinone pigment from Shiraia fruiting bodies, functions as an efficient photosensitizer for clinical photodynamic therapy. Glucose-6-phosphate dehydrogenase (G6PDH), the rate-limiting enzyme of the pentose phosphate pathway (PPP), governs carbon flux into NADPH production. This study elucidates G6PDH’s regulatory role in HA biosynthesis in Shiraia sp. S9. Bamboo polysaccharide (BPS) elicitation (100 mg/L) significantly enhanced HA production to 428.1 mg/L, 1.6-fold higher than controls after 5 days. We cloned the G6PDH gene and demonstrated that BPS upregulated its expression and activity, concomitant with increased reactive oxygen species (ROS; H₂O₂ and O₂^•−) and nitric oxide (NO) generation. ROS production was mediated by NADPH oxidase induction, while NO generation was attributed to elevated nitric oxide synthase and nitrate reductase activities. Critically, the G6PDH inhibitor glucosamine (1.0 mM) suppressed both H₂O₂ and NO production. These ROS/NO signals upregulated key HA biosynthetic (PKS, Omef) and transport (MFS) genes. Our findings establish G6PDH as a central regulator of BPS-induced HA biosynthesis via ROS/NO signaling, revealing novel metabolic crosstalk between the PPP and fungal perylenequinone biosynthesis. This work presents BPS elicitation as a biotechnological strategy for scalable HA production in Shiraia mycelium cultures. Full article

Ilex verticillata (winterberry) is a valuable ornamental shrub increasingly threatened by leaf blight, a disease that compromises its aesthetic and economic value. While fungal pathogens like Alternaria alternata are known to cause leaf blight in horticultural crops, their role in I. verticillata and the host’s defense mechanisms have not been fully characterized. Our study investigated the pathogen-host interaction by identifying the causal agent and examining the physiological and molecular defense mechanisms of I. verticillata. Through morphological and multi-locus molecular analyses (ITS, TEF1-α, G3PDH, RPB2), A. alternata was confirmed as the primary pathogen, fulfilling Koch’s postulates. Pathogenicity assays revealed distinct disease progression stages, from necrotic lesions to tissue degradation. Transcriptomic profiling uncovered dynamic host responses, with early upregulation of pattern recognition receptors (PRRs) and transcripts encoding antioxidant enzymes (SOD, CAT), followed by downregulation of metabolic pathway genes. Phytohormone analysis highlighted intricate crosstalk, with salicylic acid (SA) peaking during mid-infection and jasmonic acid (JA) rebounding later, reflecting a coordinated defense strategy. Additionally, the oxidative stress marker malondialdehyde (MDA), an indicator of membrane lipid peroxidation, surged early, indicating membrane damage, while sustained induction of antioxidant enzymes suggested adaptive responses. The key finding was distinct phytohormone crosstalk, characterized by a mid-infection SA peak followed by a late JA rebound, alongside an early oxidative burst marked by MDA accumulation and sustained antioxidant enzyme activity. These findings provide a framework for understanding I. verticillata’s defense mechanisms and offer insights for developing targeted disease management strategies, such as resistant cultivar breeding or hormone-mediated interventions. Full article

Background/Objectives: Streptococcus suis (SS), an important zoonotic pathogen, has caused significant economic losses to the global pig industry. Existing commercial vaccines for SS mainly provide effective protection against a single serotype. Due to the existence of many serotypes and their robust immune evasion capabilities, the development of multi-component subunit vaccines or multi-epitope vaccines that provide effective cross-protection against different strains of SS is a key focus of current research. Methods: We applied two-dimensional electrophoresis (2-DE) and immunoblotting to screen for candidate immunogens among the immunogenic cell wall proteins of SS. BALB/c mice were immunized intradermally with a multi-component, multi-epitope vaccine. The vaccine’s safety and immunogenicity were assessed via clinical monitoring, antibody titer detection, cytokine assays, and survival curve analyses. Results: In this study, eight immunogenic cell wall proteins (GH25, Pk, PdhA, Ldh, ExoA, Pgk, MalX, and Dnak) were successfully identified using MALDI-TOF-MS, all of which could induce high IgG antibody titers. Based on the conservation and immunoprotection demonstrated by these eight protective antigenic proteins, PdhA, Ldh, and MalX were screened to construct a multi-component subunit vaccine as a candidate vaccine for providing cross-protection against SS isolates of multiple serotypes. Challenge studies showed that mice immunized with the multi-component subunit vaccine (PdhA, Ldh, and MalX) were protected against challenges with the SS2 virulent strain ZY05719 (62.5% protection) and the SSChz virulent strain CZ130302 (75% protection). Subsequently, we utilized immunoinformatics techniques to design a novel multi-epitope vaccine (MVPLM) derived from the immunogenic proteins PdhA, Ldh, and MalX. However, challenge tests revealed that the MVPLM offered limited protection against SS. Conclusions: These data demonstrate that a multi-component subunit vaccine composed of PdhA, Ldh, and MalX proteins shows promise as a candidate universal vaccine against multiple SS serotypes. Full article

Healthcare access is a fundamental human right, yet barriers often negatively impact health, particularly in developing countries like Pakistan, where maternal mortality remains a crisis. This study aimed to identify factors influencing healthcare access barriers among married women aged 15–49 years using spatial analysis. Methods: Data were drawn from the 2017-18 Pakistan Demographic and Health Survey (PDHS), which included an unweighted sample of 8127 women. Healthcare access barriers were identified as the outcome variable. Results: A spatial analysis using ArcGIS 10.7.1 and SaTScan identified clustered distributions, with concentration areas identified in Gilgit Baltistan, Khyber Pakhtunkhwa (KPK), Federally Administered Tribal Areas (FATA), Punjab, and Balochistan. SaTScan highlighted primary clusters in FATA, Southern KPK, Northern Balochistan, and Eastern Punjab. Geographically Weighted Regression identified women who had five or more living children, respondents who did not have four or more antenatal care (ANC) visits, respondents who experienced a lower income (wealth index), respondents who did not participate in decision-making, respondents with a primary education, and respondents who accepted domestic violence as the significant predictors of healthcare access barriers. Conclusions: To improve women’s healthcare access, integrated policy interventions are needed, addressing socioeconomic disparities, strengthening national health policies, empowering women, and expanding healthcare accessibility. Strengthening health insurance and economic empowerment is crucial for achieving Sustainable Development Goals. Full article