Reduction In Viability Research Articles

Molecular insights into the physiological impact of low-frequency noise on sea slug Onchidium reevesii: Activation of p53 signaling and oxidative stress response

To investigate the regulatory role of tumor protein p53 of sea slug (Onchidium reevesii) under oxidative stress conditions, we examined the response mechanisms of O. reevesii to low-frequency noise pollution (1000 Hz) using molecular and cellular biology techniques. We successfully cloned the O. reevesii p53 gene (Orp53) from O. reevesii, obtaining a 3356 bp sequence containing a 2727 bp open reading frame (ORF). Phylogenetic analysis revealed that O. reevesii shares a close evolutionary relationship with other molluscs, including Bulinus truncatus and Elysia marginata. Expression analysis showed that Orp53 is expressed across various tissues, with the highest expression levels in the hepatopancreas. Using RNA interference (RNAi) to silence the Orp53 gene, we found that the mRNA expressions of Orp53 and its downstream apoptosis-related genes, including cytochrome C (Cyt_C), Caspase 9, and Caspase 3, were significantly suppressed until the third day of interference (P < 0.05). Moreover, sip53 treatment resulted in significant reductions in the mRNA expression and protein levels of all studied genes (P < 0.05) compared to the noise-exposed group. In addition, the low-frequency noise exposure decreased central nervous system (CNS) viability while increasing oxidative stress markers, including reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione S-transferase (GST). On the other hand, silencing Orp53 expression via siRNA resulted in significant reductions in CNS cell viability (P < 0.05). Our study establishes a molecular basis for evaluating the consequences of marine noise pollution, confirming that low-frequency noise activates the p53 signaling pathway, oxidative stress, and that p53 can regulate oxidative stress and apoptosis-related genes.

Plant-derived extracellular vesicles release combined with systemic DOX exhibits synergistic effects in 3D bioprinted triple-negative breast cancer

Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer, lacking targeted therapeutic options. Hydrogels, particularly gelatin methacrylate (GelMA), have emerged as promising materials for localized drug delivery due to their biocompatibility and tunable properties. This study investigates a dual-delivery system for enhancing the treatment efficacy of triple-negative breast cancer (TNBC) using a combination of extracellular vesicles (EVs) derived from Citrus limon L. and the chemotherapeutic drug doxorubicin (DOX). We fabricated 3D bioprinted GelMA scaffolds to achieve localized and controlled release of EVs and evaluated their synergistic effects with systemic DOX delivery on both primary and metastatic 3D TNBC models.The GelMA scaffolds, especially those with 95 % methacrylation, exhibited higher stiffness, which enhanced their sustained release. Following 48-h incubation, the combination of EVs and DOX significantly increased cytotoxicity in the primary 3D TNBC model, reducing cell viability to approximately 30 % compared to controls. This was notably more effective than treatments with DOX or EVs alone. During the extended 7-day incubation period, the combination treatment continued to show superior efficacy, with persistently high levels of ROS generation and further reduction in cell viability. In a metastatic 3D TNBC model, a significant sensitivity to the combined treatment was observed, which notably inhibited aggregate formation and migration. Importantly, EVs-embedded scaffolds promoted the proliferation of human fibroblasts, highlighting their non-toxic nature, while concurrently inhibiting TNBC cell growth. This approach provides a promising strategy to improve the treatment outcomes of TNBC by exploiting the synergistic effects of local EVs release and systemic chemotherapy.

In vitro and in vivo investigation of the inhibitory effects of Sinoporphyrin sodium-mediated Sonodynamic therapy on human oral squamous cell carcinoma

ObjectiveSonodynamic therapy (SDT) is an innovative, non-invasive approach to cancer treatment, by using low-intensity ultrasound to trigger the activation of sonosensitizers localized within cancerous cells. This current study aimed to explore the therapeutic efficacy of a new sonosensitizer, Sinoporphyrin Sodium (DVDMS), under ultrasound irradiation, against oral squamous cell carcinoma (OSCC)-derived SCC-154 cells, both in vitro and in vivo. MethodsFluorescence spectra, cytotoxicity assessments, uptake mechanisms, and subcellular distributions of DVDMS within the SCC-154 cell line were detected. Additionally, the study comprehensively assessed the antitumor effect, oxidative stress responses, apoptosis, apoptosis-related proteins, autophagic processes, and ultrastructural changes in SCC-154 cells, both in vitro and in vivo, subsequent to treatment with low-intensity ultrasound (at 1.0 MHz, 1 W/cm2 in vitro and 3 W/cm2 in vivo) in conjunction with DVDMS also being examined. ResultsThe findings indicate that SCC-154 cells exhibit heightened sensitivity to DVDMS compared to SAS and HSC-3 cell lines. Within SCC-154 cells, DVDMS primarily localizes within the mitochondria and lysosomes. DVDMS-based SDT significantly increased the intracellular levels of reactive oxygen species (ROS), induced morphological changes such as mitochondrial swelling and formation of autolysosomes, and exhibited a notable dose-dependent reduction in cell viability in vitro. Also, DVDMS-SDT demonstrated significant inhibition of xenograft growth without discernible adverse effects. Mechanistically, DVDMS-SDT upregulated Bax expression while downregulating Bcl-2 expression, which led to the Bax/Bcl-2 ratio and induced autophagy. ConclusionDVDMS-SDT triggers mitochondrial-dependent apoptosis in SCC-154 cells, unlike 5-ALA and protoporphyrin IX (PpIX). Also, the combination of DVDMS with ultrasound stimulation induces autophagy, with the onset of autophagic processes preceding apoptosis.

Mechanical Disruption by Focused Ultrasound Re-sensitizes ER+ Breast Cancer Cells to Hormone Therapy

ObjectiveTamoxifen is the most used agent to treat estrogen receptor-positive (ER+) breast cancer (BC). While it decreases the risk of cancer recurrence by 50%, many patients develop resistance to this treatment, culminating in highly aggressive disease. Tamoxifen resistance comes from the repression of ER transcriptional activity that switches the cancer cells to proliferation via nonhormonal signaling pathways. Here, we evaluate a potential strategy to overcome tamoxifen resistance by focused ultrasound (FUS), a noninvasive approach for the mechanical excitation of cancer cells. MethodsResistant and nonresistant ER+ BC cells and xenografts from patients with ER+ BC were treated with tamoxifen, FUS or their combination. The apoptosis, proliferation rate, gene expression and activity of estrogen receptor, and morphological changes were measured in treated cells and tissues. ResultsFUS caused the mechanical disruption of tamoxifen-resistant BC cells that in turn led to the upregulation of ERα-encoding gene expression and long-term re-sensitization of the cells to tamoxifen. Patient-derived xenografts treated with Tamoxifen and FUS demonstrated a significant reduction in tumor viability and proliferation and a strong structural damage to tumor cells and extracellular matrix. ConclusionFUS can improve ER+ BC treatment by re-sensitizing the cancer cells to tamoxifen.

DFT and molecular docking research on the effects of lichen metabolites

Breast cancer remains a significant public health problem worldwide and requires the investigation of new treatment methods. Lichens, symbiotic organisms rich in bioactive compounds, have emerged as promising sources of natural anticancer agents. This study investigates the anticancer effects of lichen metabolites derived from Xanthoparmelia wyomingica on breast cancer cell lines MCF-7 and MDA-MB231. High-performance liquid chromatography (HPLC) was used to measure specific lichen acids in the extract; this revealed the presence of Gyrophoric acid (GA), Norstictic acid (NA), Protocetraric acid (PA), Stictic acid (SA), and Usnic acid (UA). Cytotoxicity analysis showed significant reductions in cell viability, particularly in the MDA-MB231 cell line, with IC50 values ranging from 21.67 to 36.71 µg/mL. Molecular docking studies elucidated the interaction mechanisms between these metabolites and target proteins, highlighting NA as the most promising compound with the lowest binding energy (−9.5 kcal/mol) and inhibition constant (0.108755 μM). ADMET analysis of the molecules was then performed. Molecular electrostatic potential (MEP) and HOMO-LUMO analysis provided further information about the reactivity and electron distribution of the compounds. This study has the potential to provide novel therapeutic agents for breast cancer to tackle the increasing resistance to conventional therapies. Lichen metabolites enable the development of effective therapies in targeting aggressive subtypes such as triple-negative breast cancer with limited treatment options. This research highlights the anticancer properties of lichen metabolites and their potential in the development of innovative strategies for breast cancer treatment. The findings offer a promising avenue to address the urgent need for effective therapies in oncology.

Evaluation of Spirooxindole-3,3'-pyrrolines-incorporating Isoquinoline Motif as Antitumor, Anti-Inflammatory, Antibacterial, Antifungal, and Antioxidant Agents.

A series of novel 2-(isoquinolin-1-yl)-spiro[oxindole-3,3'-pyrrolines] were synthesized by a one-pot three-component reaction involving dimethyl acetylenedicarboxylate, 3- phenylimidazo[5,1-a]isoquinoline and N-alkylisatins in chloroform at ∼60 °C for 24 h. This study aimed at the synthesis of novel spirooxindole-3,3'-pyrrolines derivatives and in vitro evaluation of cytotoxicity affinities in cross-correlations with their antiinflammation and radical scavenging capacities. The objective of this study was to use a one-pot, three-component reaction to synthesize a novel set of spirooxindole-3,3'-pyrrolines derivatives. A novel set of spirooxindole-3,3'-pyrrolines (8a-i) was synthesized by a one-pot threecomponent reaction involving dimethyl acetylenedicarboxylate, 3-phenylimidazo[5,1-a]isoquinoline and N-alkylisatins in chloroform at ∼60 °C for 24 h. These new compounds were characterized by 1HNMR, 13C-NMR, and HRMS spectral data and screened for their antitumor, anti-inflammatory, antibacterial, antifungal, and antioxidant activities. The new synthetic spirooxindole-3,3'-pyrrolines (8a-i)-tested compounds displayed significant anti-inflammatory properties and were noncytotoxic on PDL fibroblasts. However, they lacked antioxidative-DPPH radical scavenging capabilities. Notably, Doxorubicin and cisplatin demonstrated antiproliferative effects on various cancer monolayers. Moreover, compounds 8b, 8d, 8f, 8h, and 8i exhibited pronounced viability reduction properties in colorectal and pancreatic cancer monolayers, as well as across skin, lung, prostate, and cervical adenocarcinomas, with higher cytotoxicity in mammary cancer cells MCF7 and T47D. None of the tested compounds had significant antibacterial activity against S. aureus or E. coli. However, compounds 8c, 8d, and 8f exhibited notable antifungal properties, indicating potential for further investigation. Eight new synthetic spiro[indoline-3,3-pyrroles] were prepared, characterized, and evaluated for their anti-inflammatory and cytotoxic properties. The compounds showed significant anti-inflammatory effects and promising cytotoxicity against various cancer monolayers, especially in colorectal and pancreatic cancers. Some compounds also exhibited antifungal properties. However, they did not exhibit significant antibacterial activity.

EP300-mediated H3 acetylation elevates MTHFD2 expression to reduce mitochondrial dysfunction in lipopolysaccharide-induced tubular epithelial cells.

Sepsis represents an organ dysfunction resulting from the host's maladjusted response to infection, and can give rise to acute kidney injury (AKI), which significantly increase the morbidity and mortality of septic patients. This study strived for identifying a novel therapeutic strategy for patients with sepsis-induced AKI (SI-AKI). Rat tubular epithelial NRK-52E cells were subjected to lipopolysaccharide (LPS) exposure for induction of in-vitro SI-AKI. The expressions of E1A binding protein p300 (EP300) and methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) in NRK-52E cells were assessed by western blot and qRT-PCR, and their interaction was explored by chromatin immunoprecipitation performed with antibody for H3K27 acetylation (H3K27ac). The effect of them on SI-AKI-associated mitochondrial dysfunction of tubular epithelial cells was investigated using transfection, MTT assay, TUNEL staining, 2',7'-Dichlorodihydrofluorescein diacetate probe assay, Mitosox assay, and JC-1 staining. MTHFD2 and EP300 were upregulated by LPS exposure in NRK-52E cells. LPS increased the acetylation of H3 histone in the MTHFD2 promoter region, and EP300 suppressed the effect of LPS. EP300 ablation inhibited the expression of MTHFD2. MTHFD2 overexpression antagonized LPS-induced viability reduction, apoptosis promotion, reactive oxygen species overproduction, and mitochondrial membrane potential collapse of NRK-52E cells. By contrast, MTHFD2 knockdown and EP300 ablation brought about opposite consequences. Furthermore, MTHFD2 overexpress and EP300 ablation counteracted each other's effect in LPS-exposed NRK-52E cells. EP300-mediated H3 acetylation elevates MTHFD2 expression to reduce mitochondrial dysfunction of tubular epithelial cells in SI-AKI.

Pharmacological inhibition of ezrin reduces proliferative and invasive phenotype in acute lymphoblastic leukemia cells

Ezrin (EZR) is an actin-associated protein that is often upregulated in cancers. Here we investigate the role of EZR in acute lymphoblastic leukemia (ALL) and explore the therapeutic potential of a pharmacological EZR inhibitor, NSC305787. ALL patient cohorts exhibit significantly elevated EZR mRNA levels, indicating its association with the malignant phenotype. Notably, EZR expression does not impact survival outcomes or relevant clinical-laboratory characteristics, suggesting a role in disease initiation rather than therapy response. NSC305787 induces a dose-dependent reduction in ALL cell viability, and is more potent than a related EZR inhibitor, NSC668394. NSC305787 has multiple effects on ALL cells, including apoptosis induction, clonal growth reduction, and inhibition of cell cycle progression. Importantly, it diminishes adhesiveness and invasiveness in ALL cells. Proteomics analysis highlights changes in translation, RNA catabolism, and cell cycle regulation, emphasizing the broad impact of EZR inhibition on ALL cell biology. Ex vivo assays with primary cells from acute myeloid leukemia (AML) and ALL patients demonstrate NSC305787's efficacy across a molecularly heterogeneous group, independent of risk stratification or recurrent mutations. Notably, NSC305787 shows heightened potency in ALL cells, suggesting its potential as a targeted therapy. In conclusion, our results report high EZR expression in adult ALL patients and support NSC305787 as a promising targeted therapy for ALL that should be further explored.

The study of the antiviral effect of a soft dosage form

Aim. To study the antiviral activity of a soft medicine with mangiferin against the herpes simplex virus. Materials and methods. The studies used samples of the soft dosage form developed with mangiferin, a xanthone belonging to the class of polyphenols. Mangiferin is a light yellow fine crystalline powder obtained from the leaves of the mango tree (Mangifera indica), (manufacturer Shaanxi, China). The substance contains 98.5 % of mangiferin. The antiviral activity was studied on Vero kidney cells of an African green monkey obtained from the Cell Bank of the Institute of Experimental Pathology, Oncology and Radiobiology named after R. E. Kavetsky of the National Academy of Sciences of Ukraine. The cell cultivation was performed according to standard methods. The cultivation and accumulation of the virus was carried out on Vero cell culture. The infectious titer of the virus was 7.4 log10 TCD 50/ml. The MTT modification analysis was used to study the cell viability. The antiviral activity was studied according to standard methods. The virulicidal activity against HSV-1 was determined by decreasing the cytopathic effect of the virus on cells and the virus titer using the MTT method. Results and discussion. The determination of the direct effect of the test sample on the extracellular herpes virus (virucidal effect) of an undiluted sample and samples in a dilution of 1:2.5 and 1:5 showed their ability to exhibit a significant virucidal effect against HSV-1 regardless of the dilution used and the exposure time with the virus. It was found that the decrease in the titer of the herpes virus was within 1.7-2.2 log10. When studying the ability of the test sample to influence the development of the cytopathic effect of the herpes virus on cells, it was determined that in dilutions (1:20-1:320), the sample inhibited the reproduction of the virus and the development of HSV-1 on Vero cells compared to the control virus. The greatest efficiency was found when using the sample in dilutions of 1:80-1:320; the reduction in the development of the cytopathic effect of the virus on cells was in the range of 59-64 %. When studying the infectious titer of the herpes virus synthesized de novo after the treatment with various dilutions of the test sample, it was shown that the test sample in the dilutions used did not significantly affect the synthesis of the infectious progeny of the virus, the decrease in the viral titer did not exceed 1.1 log10. Cytotoxicity assays showed that in the dilutions of 1:40 or more, the maximum reduction in cell viability was 18 %, indicating low toxicity of the sample. Conclusions. According to the results of the study, the ability of the test sample containing 5 % mangiferin to effectively inactivate the extracellular herpes simplex virus Type 1 and significantly inhibit the development of the cytopathic effect of the virus on cells has been found.

Real-time evaluation of macozinone activity against Mycobacterium tuberculosis through bacterial nanomotion analysis.

Novel drugs and improved diagnostics for Mycobacterium tuberculosis (MTB) are urgently needed and go hand in hand. We evaluated the in vitro activity of two benzothiazinone drug candidates (MCZ, PBTZ169; BTZ043) and their main metabolites against MTB using advanced nanomotion technology. The results demonstrated significant reductions in MTB viability within 7 h, indicating the potential for rapid, precise antibiotic susceptibility testing based on a phenotypic read-out in real time. PBTZ169 and H2-PBTZ169 achieved 100% separation between the susceptible H37Rv and a resistant dprE1 mutant strain NTB1. These findings support nanomotion technology's potential for faster antibiotic susceptibility testing of novel MTB drug candidates targeting the DprE1 enzyme that could reduce empirical treatment duration and antibiotic resistance selection pressure due to inaccurate treatments.

Methylglyoxal impairs human dermal fibroblast survival and migration by altering RAGE-hTERT mRNA expression in vitro

Fibroblasts are native residents in dermal layer of human skin which are important for dermal regeneration and essential during cutaneous wound healing by releasing inflammatory markers and actively migrate to close an open wound. Premature skin ageing due to methylglyoxal (MGO) has recently caught the attention considering its potential to accelerate the emergence of skin ageing signs, however previous studies were only focused in primary neonatal dermal fibroblast and NIH3t3 fibroblast cell line. Therefore, thorough investigation is required to study the impact of MGO on primary human dermal fibroblast isolated from adult subject (HDFa). In our experiments, short exposure of MGO was observed to induced significant reductions in cell viability at concentrations of 7.5, 10, 12.5, 15, and 17.5 mM (p < 0.005) after 3 hours of treatment. The cellular death of HDFa at 10, 12.5 and 15 mM of MGO were also marked by increased in intracellular ROS level, indicating the involvement of oxidative stress-induced death in these cells. We also observed enlarge scratch areas of cells exposed with 7.5 and 10 mM MGO compared to control after 26 hours, thereby suggesting a decline in cell migration and viability in this group. We propose the increased ROS as the consequence of AGE-RAGE activation which was marked by significant elevation of RAGE mRNA on cells exposed to 10 mM MGO. Our data also suggest the occurrence of DNA damage events via ROS-induced oxidation or mediated by decline in hTERT mRNA expression.

Curcumin Inhibits Oxidative Stress and Apoptosis Induced by H2O2 in Bovine Adipose-Derived Stem Cells (bADSCs)

In livestock production, oxidative stress (OS) is ubiquitous, reducing animal productivity and product quality. Hence, investigating the mechanisms of oxidative stress in livestock and inhibiting oxidative stress-induced damage is crucial. Curcumin, a plant-derived bioactive compound, exhibits antioxidant and anti-apoptotic properties. Adipose-derived stem cells (ADSCs) from animal adipose tissue are easily accessible and possess multilineage differentiation potential. Therefore, this work utilized bovine ADSCs to establish an oxidative stress model and investigated the effects of curcumin on oxidative stress and apoptosis. Firstly, bovine ADSCs were isolated and cultured from fetal calf subcutaneous adipose tissue. Their surface markers were identified by immunofluorescence, confirming the expression of CD29, CD44, CD73, CD90, CD105 and Vimentin, but not CD34, indicative of mesenchymal stem/progenitor cell characteristics. Secondly, to explore the effects of curcumin on oxidative damage and apoptosis in bovine ADSCs, an oxidative stress model was induced using H2O2. CCK-8 assays showed significantly reduced cell viability and SOD activity, along with increased malondialdehyde (MDA) and reactive oxygen species (ROS) levels, indicating successful modeling. RT-qPCR further confirmed that 500 μM of H2O2 treatment for 24 h promoted apoptosis. Herein, CCK-8 assays indicated a significant reduction in cell viability at &gt;8 μM of curcumin. Thirdly, using 4 μM and 8 μM of curcumin for pre-protection, 8 μM maintained SOD activity, reduced MDA and ROS, inhibited apoptosis-related gene changes (Bcl-2, Bax, Caspase-3), and suppressed apoptosis according to a TUNEL assay. Fourthly, curcumin’s autophagy-inducing potential was hypothesized, which was confirmed by increased LC3-II and decreased P62 expression upon co-treatment with 3-MA. 3-MA inhibited curcumin’s antioxidant and anti-apoptotic effects, suggesting that curcumin’s antioxidant and anti-apoptotic roles may involve autophagy induction. In conclusion, bovine ADSCs are abundant, easily accessible, and multipotent, making them suitable for in vitro expansion. Curcumin alleviated H2O2-induced oxidative stress in bovine ADSCs, with curcumin also inhibiting apoptosis, likely through autophagy induction. This study validates the protective role of curcumin in bovine ADSCs, with potential applications in livestock production.

In Vitro Apoptotic and Antiproliferative Activity of Hypericum Perforatum Extract on Human Osteosarcoma Cell Line.

Hypericum perforatum (HP) has been widely used as an alternative medicine due to its active pharmacological properties. While the antiproliferative effects of components such as hypericin and hyperforin have been demonstrated in malignant cell lines, most studies have focused on the pharmacological properties of the HP extract itself. Recent research has indicated that HP and its active substances possess anticancer activities; however, there is a lack of studies examining its effects on osteosarcoma. In addition, HP has demonstrated the ability to mitigate the toxicity of several drugs, including chemotherapeutic agents. Hence, the primary objective of this study was to explore the potential anticancer properties of HP in relation to osteosarcoma cells. MG-63 human osteosarcoma cells were cultured and treated with HP extract. Apoptotic factors were analyzed using ELISA, while cell viability was assessed using the MTT test. The results revealed a significant increase in the activities of proapoptotic proteins GRP78, Wee1, apoptosis-inducing factor (AIF), GADD153, Bax, and cleaved caspase-3 in MG-63 osteosarcoma cells after 48 hours of treatment with HP at a concentration of 0.8%. Conversely, the activity of Bcl-2, an antiapoptotic protein, significantly decreased. Moreover, HP extract demonstrated a dose-dependent reduction in cell viability in MG-63 cells. In conclusion, HP extract induces apoptosis in MG-63 osteosarcoma cells by upregulating the expressions of proapoptotic proteins GRP78, Wee1, AIF, GADD153, Bax, and cleaved caspase-3. This study will assist researchers in understanding the importance of alternative treatments using HP in the context of human osteosarcoma therapy, which many researchers are currently unaware of.

Knockout of RIG-I in HEK293 cells by CRISPR/Cas9

We knocked out the retinoic acid-inducible gene I (RIG-I) in HEK293 cells via CRISPR/Cas9 to reveal the effects of RIG-I knockout on the key factors in the type I interferon signaling pathway. Three single guide RNAs (sgRNAs) targeting RIG-I were designed, and the recombination vectors were constructed on the basis of the pX459 vector and used to transfect HEK293 cells, which were screened by puromycin subsequently. Furthermore, a mimic of virus, poly I: C, was used to transfect the cells screened out. RIG-I knockout was checked by sequencing, real-time quantitative PCR, Western blotting, and immunofluorescence assay. Meanwhile, the expression levels of key factors of type I interferon signaling pathway such as melanoma differentiation-associated gene 5 (MDA5), interferonβ1 (IFNβ1), and nuclear factor-kappa B p65 [NF-κB(p65)], as well as cell viability, were determined. The results showed that two HEK293 cell lines (S1 and S3) with RIG-I knockout were obtained, which exhibited lower mRNA and protein levels of RIG-I than the wild type HEK293 cells (P < 0.05). The mRNA levels of MDA5 and IFNβ1 in S1 and S3 cells and the protein level of NF-κB(p65) in S3 cells were lower than those in the wild type (P < 0.05). More extranuclear NF-κB(p65) protein was detected in S1 cells than in the wild type after transfection with poly I: C. Plus, the wild-type and S1 cells transfected with poly I: C for 48 h showcased reduced viability (P < 0.05), while S3 cells did not display the reduction in cell viability. In summary, the present study obtained two HEK293 cell lines with RIG-I knockout via CRISPR/Cas9, which provided a stable cell model for exploring the mechanism of type I interferon signaling pathway.

3D in vitro modelling of post-partum cardiovascular health reveals unique characteristics and signatures following hypertensive disorders in pregnancy

BackgroundHypertensive disorders of pregnancy (HDP) affect 2–8% of pregnancies and are associated postpartum with increased cardiovascular disease (CVD) risk, although mechanisms are poorly understood.MethodsHuman induced pluripotent stem cells (iPSC)-derived cardiomyocytes, cardiac fibroblasts and coronary artery endothelial cells were cocultured to form cardiac spheroids (CSs) in collagen type-1 hydrogels containing 10% patient plasma collected five years postpartum [n = 5 per group: normotensive control, gestational hypertension (GH) and preeclampsia (PE)]. Plasma-treated CSs were assessed for cell viability and contractile function and subjected to immunofluorescence staining and imaging. A quantitative proteomic analysis of plasma samples was conducted (controls n = 21; GH n = 5; PE n = 12).ResultsContraction frequency (CF) was increased in PE-treated CSs (CF: 45.5 ± 3.4 contractions/minute, p < 0.001) and GH-treated CSs (CF: 45.7 ± 4.0 contractions/minute, p < 0.001), compared to controls (CF = 21.8 ± 2.6 contractions/min). Only PE-treated CSs presented significantly increased fractional shortening (FS) % (9.95 ± 1.8%, p < 0.05) compared to controls (3.7 ± 1.1%). GH-treated CSs showed a reduction in cell viability (p < 0.05) and an increase in α-SMA expression (p < 0.05). Proteomics analyses identified twenty differentially abundant proteins, with hemoglobin A2 being the only protein perturbed in both GH and PE versus control plasma (p < 0.05).ConclusionsThe innovative patient-relevant CS platforms led to the discovery of biomarkers/targets linked to cell death signaling and cardiac remodeling in GH-induced CVD and vascular/endothelial cell dysfunction in PE-induced CVD.

Does Combined Treatment with Tranexamic Acid and Vancomycin Affect Human Chondrocytes In Vitro?

Background: The aim of our study was to examine the combined effects of tranexamic acid (TXA) and vancomycin powder (VP) on chondrocytes in vitro. Despite the use of TXA and VP being linked to a reduced risk of extensive postoperative blood loss and periprosthetic joint infections (PJIs) in TKA, the possible cytotoxic side effects on periarticular cell types remain unclear. Methods: Human chondrocytes were harvested from hyaline cartilage and expanded in monolayer culture before being simultaneously exposed to different concentrations of TXA and VP for varying exposure times. Cell viability and proliferation were assessed using an ATP assay and an Annexin 5 assay, respectively, while changes in the relative expression of chondrogenic marker genes were examined using semiquantitative RT-PCR. Results: The simultaneous exposure of chondrocytes to TXA and VP for more than 48 h led to a reduction in both cell viability and proliferation rates. When exposing chondrocytes to the lowest examined concentrations of both TXA (10 mg/mL) and VP (3 mg/mL), the observed effects were delayed until 96 h. However, our study found no dependencies of the observed effects on the concentrations tested. Further, we found no effects on the expression of chondrogenic marker genes. Conclusions: Consequently, limiting the exposure time of chondrocytes to TXA and VP in an in vitro setting to 24 h may be considered safe and could help to further improve the understanding of the safe use of substances in vivo. However, further in vitro research is required to develop a comprehensive understanding of the effects of both VP and TXA on important periarticular cell types in TKA, including chondrocytes, osteocytes, and tenocytes.

End of the Season Blues: Senescence and Reproductive Trade-Offs in Male Scorpions

Seasonal reproductive dynamics and senescence have profound impacts on male fertility, yet these processes remain understudied in scorpions. This study investigates how reproductive parameters—such as testicular mass, sperm quantity, and viability—change over the course of the reproductive season in Urophonius achalensis males. We found that early-season males exhibited higher sperm quality and testicular mass compared to their older counterparts, suggesting that senescence, rather than reproductive effort, drives the decline in sperm viability. Interestingly, isolated males accumulated more sperm without a reduction in viability, highlighting potential mechanisms that preserve sperm quality despite senescence. Body condition also played a key role, with males in better condition showing higher sperm counts but facing a sharper decline in sperm viability as the season progressed, suggesting trade-offs between early reproductive success and later-life reproductive costs. Our findings offer novel insights into the reproductive strategies of scorpions, emphasizing the interplay between senescence, environmental stressors, and reproductive investment. This work provides new insights into the reproductive biology of scorpions, with broader implications for understanding the role of senescence and mating systems in shaping reproductive strategies.

Lidocaine Induces Neurotoxicity by Activating the CaMKII and p38 MAPK Signaling Pathways through an Increase in Intracellular Calcium Ions

Background: Lidocaine is extensively utilized as an anesthetic in clinical settings; however, it has demonstrated significant neurotoxicity when administered for spinal anesthesia. The specific mechanisms underlying lidocaine-induced neurotoxicity are poorly understood. Objective: This study aimed to investigate the mechanisms through which lidocaine induces neurotoxicity, focusing on its effects on intracellular calcium release and the activation of CaMKⅡ and MAPKs pathways, as well as to evaluate the potential protective effects of cilnidipine. Methods: The investigation has employed both in vitro cell models and in vivo mouse models to conduct the experiments. Neuronal cell viability has been assessed following lidocaine treatment, and neurological function has been evaluated in mice after intrathecal injection of lidocaine. Intracellular calcium levels, CaMKⅡ activation, and the phosphorylation of p38 and p65 have been measured in cultured hippocampal neuronal cells and mouse brain tissues. The effects of the calcium channel blocker cilnidipine on these parameters have also been examined. Results: Lidocaine treatment led to a reduction in cell viability in cultured neuronal cells and induced neurological dysfunction in mice. It increased intracellular Ca2+ levels and activated CaMKⅡ in both cultured neuronal cells and mouse brain tissues. Lidocaine also elevated the phosphorylation levels of p38 and p65 in neuronal cells. These effects have been suppressed by cilnidipine, indicating a calcium-dependent mechanism. Conclusion: This study suggests that lidocaine induces neurotoxicity through a calcium-dependent activation of CaMKⅡ and MAPK pathways, leading to neuronal apoptosis and dysfunction. Cilnidipine has been found to exhibit promise as a protective agent against lidocaine-induced neurotoxicity.

Co-exposure of ferruginous components of subway particles with lipopolysaccharide impairs vascular function: A comparative study with ambient particulate matter

Several empirical studies have linked subway and ambient particle exposure to toxicity, pro-inflammatory responses, and vascular dysfunction. However, the health effects of pollutants generated from varying sources, particularly when combined with lipopolysaccharide (LPS), are still unexplored. Therefore, the aim of this study was to investigate the characteristic health effects of iron oxide particles (the main components of subway particles) in comparison with urban aerosols (UA) and vehicle exhaust particles (VEP), alone and in combination with LPS. This study revealed that iron oxides caused a more significant reduction in human umbilical vein endothelial cell viability, increased lactate dehydrogenase release, and decreased the production of plasminogen activator inhibitor-1, a fibrinolytic modulator, and endothelin-1, a vasoconstrictor, compared to those by VEP and UA at marginally toxic and toxic concentrations. While VEP and UA induced an increase in interleukin (IL)-6 production, iron oxides, particularly Fe3O4, increased IL-8 production at slightly toxic and non-cytotoxic concentrations. In addition, co-exposure of all particles and LPS at non-cytotoxic concentrations promoted pro-inflammatory cytokine (IL-6 and IL-8) production relative to exposure to the particles alone. Interestingly, the tendency towards either coagulation or fibrinolytic conditions was dependent on the concentration of exposed particles at the same LPS concentration. Furthermore, increases in inflammation, neutrophil and lymphocyte recruitment around blood vessels, and edema were observed in murine lungs exposed to a combination of iron oxides and LPS compared to those in mice exposed to iron oxide alone. Thus, iron oxide-rich subway particulate poses more health risks than outdoor ambient particles since they can significantly impair endothelial function, particularly through gross cellular and vascular homeostatic protein damage, and induce exacerbated inflammatory responses during co-exposure. These findings provide novel empirical evidence for epidemiological studies seeking mechanisms responsible for the observed health impact of transport- and occupational-related exposures on vascular dysfunction.

Polyphyllin VII enhances the sensitivity of endometrial carcinoma cells to medroxyprogesterone acetate through upregulating miR‑33a‑5p expression.

Endometrial carcinoma (EC) often exhibits resistance to hormone therapies, such as medroxyprogesterone acetate (MPA), highlighting the need for novel strategies to enhance therapeutic efficacy. The present study aimed to investigate the effects of polyphyllin VII (PPVII) on the efficacy of MPA in EC, focusing on the regulatory role of microRNA (miR)-33a-5p. Briefly, an MPA-resistant Ishikawa cell line (Ishikawa/MPA-R), maintained with 10 µM MPA, was established and transfected with negative control (NC) and miR-33a-5p inhibitors. Following treatment with PPVII and MPA, the proliferation capacity and apoptosis levels of the Ishikawa and Ishikawa/MPA-R cells were evaluated using reverse transcription-quantitative polymerase chain reaction, MTT assay, clonogenic assay, flow cytometry, western blotting and dual-luciferase assay. Next, the expression levels of miR-33a-5p and F-box and leucine rich repeat protein 16 (FBXL16) were measured, and the regulatory relationship between miR-33a-5p and FBXL16 was analyzed. Significant reductions in cell viability were observed in all groups following treatment with increased concentrations of MPA and PPVII, with the greatest effect observed in the combined MPA + PPVII group (P<0.01). The apoptosis levels of the Ishikawa/MPA-R cells were significantly increased in all drug treatment groups, particularly in the MPA + PPVII group (P<0.05). PPVII treatment significantly increased the expression level of miR-33a-5p in Ishikawa/MPA-R cells (P<0.01). In the PPVII + miR-33a-5p inhibitor group, the Ishikawa/MPA-R cells exhibited an upregulation in the viability (P<0.01), colony formation ability (P<0.01), proportion in the G1 phase (P<0.05) and the protein expression levels of cyclin D1 (P<0.01) and cyclin-dependent kinase 4 (P<0.01), and a reduction in the miR-33a-5p expression (P<0.01), apoptosis levels (P<0.05), proportion in the S (P<0.05) and G2 phases and the levels of Bcl-2-associated X protein (P<0.001). The FBXL16 protein expression in Ishikawa/MPA-R cells was significantly higher compared with Ishikawa cells, and the mRNA and protein expression levels of FBXL16 were markedly elevated in the PPVII + miR-33a-5p inhibitor group compared with the PPVII + NC group (P<0.01). These findings suggested that PPVII upregulated the expression of miR-33a-5p, enhanced the sensitivity of EC cells to MPA and potentially exerted anticancer effects in EC through the synergistic action of the miR-33a-5p/FBXL16 axis in combination with MPA.