Number Of Dead Cells Research Articles

The Effect of Zeolite Zinc on Memory Performance and Hippocampal Cell Death in a Rat Model of Alzheimer's-like Disease Induced by Aβ1-42.

Alzheimer's disease (AD) is one of the most common neurodegenerative disorders, characterized by the slow and progressive loss of brain structure and function, primarily affecting older individuals. Evidence has shown that disruption of zinc homeostasis in the brain contributes to synaptic dysfunction, as well as impairments in learning and memory. In this study, we evaluated the effect of zeolite zinc on memory performance and hippocampal cell death in a rat model of Alzheimer's disease (AD) induced by intracerebroventricular administration of Aβ1-42. We employed the Morris water maze, shuttle box, and open field tests to assess spatial memory, passive avoidance memory, and anxiety-like behavior, respectively. P-Tau and the amyloid precursor protein (APP) expression, along with hippocampal cell death, were also evaluated. Both Aβ1-42 and zeolite zinc were injected intracerebroventricularly. The results showed that zeolite zinc partially reversed Aβ1-42-induced impairments in memory performance and mitigated the effects of Aβ1-42 on locomotor activity, although it did not fully restore baseline levels. In addition, Aβ1-42 increased the expression of APP and P-Tau, as well as the number of dead cells, whereas zeolite zinc reduced these effects. In conclusion, our findings suggest that while zeolite zinc plays a role in modulating the pathophysiology of AD, its therapeutic effects only partially reverse the progression or symptoms of AD, indicating the need for further investigation into optimal dosing or combination therapies.

Apoptosis-induced exosomes from human exfoliated deciduous teeth enhance angiogenesis in human umbilical vein endothelial cells

Exosomes derived from the stem cells of human exfoliated deciduous teeth (SHED) hold promise for tissue regeneration. Apoptotic cells release a variety of extracellular vesicles that affect intercellular communication. This study aimed to investigate the angiogenic effects of SHED-derived exosomes modified via apoptosis induction on human umbilical vein endothelial cells (HUVECs). Apoptosis was induced in SHED via serum starvation for 3 weeks and confirmed by the upregulation of the apoptotic genes, caspase 3 and 9, and via annexin V staining. The apoptotic SHED-derived exosomes were isolated, characterized, and subjected to proteomic analysis. In vitro experiments were performed to assess the effects of apoptotic SHED exosomes on the proliferation, migration, and tube formation of HUVECs. The apoptosis-induced SHED showed increased cell viability and decreased numbers of dead cells compared with those of conventional cultures while retaining their identity as mesenchymal stem cells positive for CD44, CD73, and CD90. The apoptotic SHED-derived exosomes exhibited characteristic features, such as standard size, cup-shaped morphology, and positive staining, for exosomal markers CD9, CD63, and CD81. Proteins associated with apoptosis, programmed cell death, and cellular senescence were downregulated in the apoptotic SHED exosomes, whereas those associated with extracellular matrix organization were upregulated, indicating positive angiogenesis. HUVECs treated with apoptotic SHED exosomes exhibited significantly enhanced proliferation and migration compared with those treated with normal SHED exosomes. The mesh-like structures in the apoptotic SHED exosomes exhibited significantly increased signs of angiogenesis. The findings of this study provide new insights into the potential use of apoptotic SHED-derived exosomes in regenerative medicine.

Arbuscular mycorrhizal symbiosis enhances microbial contribution to mineral-associated organic carbon persistence in the soil: Insights from soil microbial community and microbial necromass carbon

The formation of mineral-associated organic carbon (MAOC), a critical soil fraction related to soil organic carbon (SOC) sequestration, is significantly influenced by microbial processes. Yet, how arbuscular mycorrhizal (AM) symbiosis impacts MAOC accumulation in rhizosphere soils through its effects on microbial contribution remains poorly understood. Here, we assessed the microbial community of different soil samples using 16S rRNA sequencing and found that AM symbiosis influenced the soil microbial community composition and enhanced soil microbial community functions related to carbon (C) degradation. The numbers of live and dead cells (all cells in the soil) were highest in AM-RS (rhizosphere soils in the AMF treatment) based on confocal laser scanning microscopy (CLSM) images. Meanwhile, AM symbiosis increased the proportions of apoptotic cells (19.02%) and necrotic cells (12.12%) in rhizosphere soils. Using soil amino sugars as microbial biomarkers, the concentrations of bacterial necromass C (188.01 g kg-1 OC) and fungal necromass C (392.19 g kg-1 OC) were highest in the MAOC fraction in AM-RS. Additionally, the MAOC content (17.20 g kg-1 soil) and proportion (42.06%) were highest in AM-RS. This research illustrates two primary mechanisms by which AM symbiosis enhances MAOC accumulation: (1) by altering the microbial community composition and function related to C degradation; and (2) by promoting the input of microbial necromass C, especially fungal necromass C. This research broadens our horizons to understand the mechanism of microbial contribution to MAOC accumulation stimulated by AM symbiosis in rhizosphere soils and provides management practices for AMF application to SOC sequestration.

Stress-experienced monocytes/macrophages lose anti-inflammatory function via β2-adrenergic receptor in skin allergic inflammation

Psychological stress can exacerbate the development of allergies; however, the underlying mechanisms remain poorly understood. IgE-mediated cutaneous allergic inflammation (IgE-CAI) is a basophil-dependent skin allergy with eosinophil infiltration at inflammatory sites. Its resolution involves anti-inflammatory programmed death ligand 2 (PD-L2)+ macrophages. This study sought to elucidate the cellular and molecular mechanisms by which psychological stress exacerbates IgE-CAI. Neural tissue involved in stress-induced IgE-CAI exacerbation was identified by performing denervation and brain destruction experiments in mice. Immune cell transplantation, RNA sequencing, flow cytometry, and ELISA were used to identify and characterize immune cells with stress-altered functioning, followed by identification of key factors involved in IgE-CAI exacerbation. Stress-induced exacerbation of IgE-CAI was found to be sympathetic and β2-adrenergic receptor (Adrb2)-dependent. Adoptive transfer experiments revealed that stress diminished the anti-inflammatory functions of PD-L2+ macrophages through Adrb2, exacerbating the inflammation. RNA sequencing analysis indicated that PD-L2+ macrophages in stressed mice exhibit reduced expression of efferocytosis-related genes, including Gas6 and MerTK. Consequently, the efferocytic capacity of these macrophages decreased, resulting in increased numbers of dead cells in the lesions. The exacerbation and upregulation of Ccl24 expression in IgE-CAI skin lesions were countered by a caspase-1 inhibitor. Psychological stress diminishes the efferocytotic capacity of PD-L2+ macrophages, causing an accumulation of dead cells. This, in turn, heightens eosinophil infiltration through caspase-1-dependent production of Ccl24, exacerbating IgE-CAI.

Lysate of bovine adipose-derived stem cells accelerates in-vitro development and increases cryotolerance through reduced content of lipid in the in vitro fertilized embryos

Mesenchymal stem cells such as adipose-derived stem cells (ADSCs) are known to secrete factors that stimulate cell division and promote regeneration in neighboring cells. While conditioned medium from stem cells has been used in blastocyst production, no studies have examined the use of cell lysates. In this study we investigated the effects of adding ADSC lysate to in vitro culture (IVC) medium. ADSCs and fibroblasts were isolated from bovine adipose tissue and auricular tissue, respectively, and their lysates were prepared by freeze-thaw disruption. ADSC lysate was added to synthetic oviductal fluid medium. The effects on cleavage, blastocyst development rates, cell numbers, cryotolerance, gene expression (POU5F1, BAX, IGF1R, IGF2R, SOD2), lipid content, and membrane integrity were evaluated according to the experimental design. In Expt. 1, the comparison involved adding ADSC or fibroblast lysate alongside the control group. The total blastocyst rate increased when ADSC lysate was introduced (ADSCs: 40.1 %, fibroblasts: 33.1 %, control: 27.3 %). However, there were no significant differences in the number of trophoblast cells or in the inner cell mass. Experiment 2 confirmed that this increase in blastocyst development was not due to the solvent, PBS(−). In Expt. 3, addition of 10 % fetal calf serum (FCS) or ADSC lysate increased the total blastocyst rate compared to the control (control, 26.2 %; 10 % FCS, 43.4 %; 1 % ADSC lysate, 34.2 %; 10 % ADSC lysate, 48.1 %). After freezing and thawing, the survival and hatching rates of embryos with FCS were significantly lower than those of the control as well as those with added ADSC lysate. In Expt. 4, the addition of ADSC lysate or FCS had no significant effect on gene expression in blastocysts compared to control. However, the addition of FCS significantly increased the gray intensity, indicating higher lipid content compared to the control, with a significant increase in the number of dead cells in the blastocyst. These results indicate that the addition of ADSC lysate to the IVC medium accelerates bovine blastocyst development and that its 10 % addition, corresponding to 1 × 105 cells/mL, is as effective as 10 % FCS without a decrease in cryotolerance due to the increased lipid content.

MiRNA506 Activates Sphk1 Binding with Sirt1 to Inhibit Brain Injury After Intracerebral Hemorrhage via PI3K/AKT Signaling Pathway.

Intracerebral hemorrhage (ICH) is an acute neurological disorder characterized by high mortality and disability rates. Previous studies have shown that 75% of patients who survive ICH experience varying degrees of neurological deficits. Sphk1 has been implicated in a multitude of phylogenetic processes, including innate immunity and cell proliferation. An in vivo rat model of ICH and an in vitro model of neuronal oxyhemoglobin (OxyHb) were constructed. The expression level of Sphk1 was assessed using western blotting and immunofluorescence, whereas cell death following ICH was evaluated using fluoro-Jade B and terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Immunofluorescence facilitated the examination of microglial phenotypic alterations, while enzyme-linked immunosorbent assays were used to determine the concentrations of inflammatory markers. Behavioral assays were employed to assess the overall behavioral modifications of animals. Neuronal Sphk1/Sirt1 protein levels gradually increased following the induction of ICH. Elevated Sphk1 expression resulted in increased levels of anti-inflammatory microglia and reduced levels of pro-inflammatory factors. In contrast, suppression of Sphk1 expression resulted in an increased number of dead cells, thereby exacerbating neurological deficits. In vitro findings indicated that the levels of phosphorylated PI3K and AKT proteins increased in conjunction with Sphk1 expression. This study established that after ICH, Sphk1 interacts with Sirt1 to mitigate neuroinflammation, cell death, oxidative stress, and brain edema via the PI3K/AKT signaling pathway. Augmenting expression of Sphk1 significantly can ameliorate neurological impairments induced by ICH, offering novel targets and perspectives for therapeutic interventions in ICH treatment.

Investigation of the cytotoxic effect of current dentine bonding agents on human dental pulp cells

BackgroundAn ideal aesthetic restorative material should be attached to the tooth tissues by adhesion, have a smooth surface as possible, should not cause toxic reactions in the pulp and discoloration and microleakage. This study aims at comparatively assess the cytotoxicity of current adhesive systems on human dental pulp cells.Materials and methodsThe adequate density of human pulp cells was observed from the ready cell line. The passaging was performed and the 3rd passage cells were selected. Adhesive systems and MTA were used on the cultures. Trypan blue staining was conducted on the cells at the 1st, 2nd, 3rd days and a count of live and dead cells using a light microscope. The dead cells whose membrane integrity was impaired by staining with trypan blue and the viability rate was determined using live and dead cell numbers. Data analysis was performed using IBM SPSS Statistics 22.ResultsA significant difference in vialibity rates between adhesive systems was observed on the first day. No significant statistical differences were observed on the 2nd and 3rd days (p < 0.05).ConclusionFuturabond M showed similar biocompatibility with MTA on human pulp cells and it can be applied in cavities with 1–1.5 mm hard tissue between pulp and dentine.

Spray-dried cyclophosphamide-loaded polyhydroxyalkanoate microparticles: design and characterization

Background and purpose: Cyclophosphamide (CP) is a widely used antitumor and immunosuppressive drug, but it is highly cytotoxic and has carcinogenic and teratogenic potential. To reduce adverse effects of CP therapy and the frequency of its administration, the microencapsulation of CP into biodegradable polymeric matrices can be performed. However, according to the literature, only a few polymers were found suitable to encapsulate CP and achieve its’ sustained release. Experimental approach: In this research, spray-dried cyclophosphamide-loaded poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) microparticles were prepared and characterized in terms of their average hydrodynamic diameter, polydispersity index, surface morphology, zeta potential, encapsulation efficiency, drug loading, thermal properties and cytotoxicity against 3T3 cells. Key results: The obtained CP-loaded microparticles had a regular spherical shape, uniform size distribution with an average diameter of 4.21±0.04 μm and zeta potential of -34.2±0.2 mV. The encap­sulation of cyclophosphamide into the PHBV matrix led to a decrease in melting and degradation tempe­ratures and an increase in diameter, glass transition and cold crystallization temperatures compared to blank microparticles. Moreover, microencapsulation of cyclophosphamide lowered its cytotoxicity compared to the pure drug: the number of dead cells in the culture decreased by 28 %, while their metabolic activity increased by 20 %. The cumulative in vitro drug release studies showed a gradual release of CP up to 18 days, so the obtained microparticle formulation can be used as a sustained-release cyclophosphamide delivery system. Conclusion: In this research, a novel cyclophosphamide-loaded platform based on PHBV micro­particles was established and characterized. Overall, this study offers promising prospects for cancer therapy in the future.

Proposal and performance evaluation of a new parallel plate continuous cell separation device using dielectrophoresis.

Along with the rapid development of cellular biological research in recent years, there has been an urgent need for a high-speed, high-precision method of separating target cells from a highly heterogeneous cell population. Among the various cell separation technologies proposed so far, dielectrophoresis (DEP)-based approaches have shown particular promise because they are noninvasive to cells. We have developed a new DEP-based device to separate large numbers of live and dead cells of the human mammary cell line MCF10A. In this study, we validated the separation performance of this device. The results showed the successful separation of a higher percentage of cells than in previous studies, with a separation efficiency higher than 90%. In the past, there have been no confirmed cases in which a separation rate of over 90% and high-speed processing of a large number of cells were simultaneously achieved. It was shown that the proposed device can process large numbers of cells at high speed and with high accuracy.

SRPK Inhibitors Reduce the Phosphorylation and Translocation of SR Protein Splicing Factors, thereby Correcting BIN1, MCL-1 and BCL2 Splicing Errors and Enabling Apoptosis of Cholangiocarcinoma Cells.

Cholangiocarcinoma (CCA) is a malignancy of the bile duct epithelium that is commonly found in the Thai population. CCA has poor prognosis and a low survival rate due to the lack of early diagnosis methods and the limited effectiveness of current treatments. A number of oncogenic spliced-transcripts resulting from mRNA splicing errors have been reported in CCA, and aberrant mRNA splicing is suspected to be a key driver of this cancer type. The hyperphosphorylation of serine/arginine rich-splicing factors (SRSFs) by serine/arginine protein kinases (SRPKs) causes them to translocate to the nucleus where they facilitate gene splicing errors that generate cancer-related mRNA/protein isoforms. The correlation between SRPK expression and the survival of CCA patients was analyzed using data from The Cancer Genome Atlas (TCGA) dataset. The effect of SRPK inhibitors (SRPIN340 and SPHINX31) on two CCA cell lines (KKU-213A and TFK-1) was also investigated. The induction of cell death was studied by Calcein-AM/PI staining, AnnexinV/7AAD staining, immunofluorescence (IF), and Western blotting (WB). The phosphorylation and nuclear translocation of SRSFs was tracked by WB and IF, and the repair of splicing errors was examined by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). High levels of SRPK1 and SRPK2 transcripts, and in particular SRPK1, correlated with shorter survival in CCA patients. SRPIN340 and SPHINX31 increased the number of dead and apoptotic cells in a dose-dependent manner. CCA also showed diffuse expression of cytoplasmic cytochrome C and upregulation of cleaved caspase-3. Moreover, SRSFs showed low levels of phosphorylation, resulting in the accumulation of cytoplasmic SRSF1. To link these phenotypes with aberrant gene splicing, the apoptosis-associated genes Bridging Integrator 1 (BIN1), Myeloid cell leukemia factor 1 (MCL-1) and B-cell lymphoma 2 (BCL2) were selected for further investigation. Treatment with SRPIN340 and SPHINX31 decreased anti-apoptotic BIN1+12A and increased pro-apoptotic MCL-1S and BCL-xS. The SRPK inhibitors SRPIN340 and SPHINX31 can suppress the phosphorylation of SRSFs and their nuclear translocation, thereby producing BIN1, MCL-1 and BCL2 isoforms that favor apoptosis and facilitate CCA cell death.

Novel Diclofenac-NO Donor With High Affinity for Human Serum Albumin Induces Endoplasmic Reticulum Stress-mediated Cell Death in Human Pancreatic Cancer Cells.

Nitric oxide (NO) has various physiological activities. In this study, diclofenac (DF) which has a high affinity for human serum albumin (HSA) was nitrosylated to a novel NO donor (NDF). The cytotoxic effects and the mechanism of NDF were investigated. Binding experiments of NDF to HSA were performed by the ultrafiltration method. NO was measured by the Griess method. The number of dead cells were measured using annexin V. Apoptosis and endoplasmic reticulum stress were evaluated by western blotting. NDF competitively inhibits the binding of DF to HSA, suggesting that NDF and DF have equivalent binding characteristics. NDF rapidly released NOx after being dissolved. At 200 μM, NDF induced cell death in human pancreatic cancer cells. Western blotting showed that NDF promoted the cleavage of PARP, caspase-3, and caspase-7. Inhibitors of caspase-1 and caspase-9 significantly suppressed NDF-induced cell death, as did a non-specific caspase inhibitor (Z-VAD). In addition, NDF significantly increased the expression of the endoplasmic reticulum stress marker, CHOP. NDF induces apoptotic cell death by causing endoplasmic reticulum stress. The findings of this study suggest that NDF may become a promising compound for the treatment of pancreatic cancer.

Intracellular hydrogen peroxide’s effect on quality parameters of rooster sperm in freeze/thaw cycle

Relevance. One of the factors decreasing total motility after cryopreservation rooster’s sperm is influence of reactive oxygen species. Morphological and biochemical features of avian spermatozoa structure, which make them more susceptible to cryopreservation process compared to mammalian spermatozoa, may also be the reason why avian spermatozoa are more susceptible to oxidative stress.Methods. The purpose of the study is to trace the change in the level of intracellular reactive oxygen species, in particular hydrogen peroxide (H2O2), during the cryopreservation of rooster sperm, to assess its effect on the quality and viability of sperm.Results. A negative correlation (r = -0.68, p &lt; 0.05) was found between the intracellular level of hydrogen peroxide and the number of dead cells in native sperm. In the frozen (thawed) seed, a weak relationship was observed between these indicators (r = -0.10). There was a significant effect of the level of intracellular hydrogen peroxide in freshly obtained ejaculates on the overall mobility of frozen (thawed) semen (r = -0.65, p &lt; 0.05). This allows us to assume that, similarly to mammalian spermatozoa, the production by cells of an increased amount of reactive oxygen species (H2O2) in the freezing (thawing) cycle negatively affects the functional status of mitochondria, which, as is known, are the main source of energy for the sperm, ensuring the operation of the kinetic apparatus of the sperm and its general mobility. Data were obtained on the individual variability of the hydrogen peroxide content in the sperm of roosters at the age of 61 weeks in the freezing (thawing) cycle, allowing the selection of roosters according to this indicator

Features of the Microalgae and Cyanobacteria Growth in the Flue Gas Atmosphere with Different CO2 Concentrations

Nowadays, it is important to create the optimal technology for the absorption of flue gases with high CO2 content. In this regard, the aim of the investigation is to study the five different microalgae strains (Chlorella vulgaris, Chlorella ellipsoidea, Elliptochoris subsphaerica, Gloeotila pulchra, and Arthrospira platensis) under the influence of flue gases. The cultivation of microalgae was carried out in the atmosphere of flue gases with a gas flow rate of approximately 1 L·min−1 at high CO2 concentrations (3, 6, or 8%—from lower to higher concentrations), under continuous (24 h·d−1) illumination intensity of 200 µmol quanta·m−2·s−1 and a constant temperature of 27 ± 1 °C. The duration of the experiments was 12 days. Chlorella vulgaris and Chlorella ellipsoidea demonstrated the highest biomass growth rate at CO2 = 6% (0.79 and 0.74 g·L−1·d−1, respectively). The lowest growth rate (0.21 g·L−1·d−1) was achieved for Arthrospira platensis at CO2 = 3 and 6%. There was no significant drop in pH in the entire series of experiments. The results of microscopy showed a lack or a minimal number of dead cells in the strains under selected conditions. The obtained results can be used for further development of CO2 capture and storage technologies.

Novel indole Schiff base β-diiminato compound as an anti-cancer agent against triple-negative breast cancer: In vitro anticancer activity evaluation and in vivo acute toxicity study

Breast cancer is the most prevalent cancer among women globally, with triple-negative breast cancer (TNBC) associated with poor prognosis and low five-year survival rates. Schiff base compounds, known for their extensive pharmacological activities, have garnered significant attention in cancer drug research. This study aimed to evaluate the anticancer potential of a novel β-diiminato compound and elucidate its mechanism of action. The compound’s effect on cell viability was assessed using MTT assays in breast cancer cell lines including MCF-7 and MDA-MB-231. Cytotoxic effects were further analyzed using trypan blue exclusion and lactate dehydrogenase (LDH) release assays. In order to assess the mechanism of inhibitory activity and mode of cell death induced by this compound, flow cytometry of cell cycle distribution and apoptosis analysis were carried out. Apoptosis incidence was initially assessed through cell and nuclear morphological changes (Hoechst 33342/Propidium iodide (PI) staining) and further confirmed by Annexin V/PI staining and flow cytometry analysis. In addition, the effect of this compound on the disruption of mitochondrial membrane potential (MMP) and generation of the reactive oxygen species (ROS) was determined using the JC-1 indicator and DCFDA dye, respectively. The results demonstrated that the 24 h treatment with β-diiminato compound significantly suppressed the viability of MDA-MB-231 and MCF-7 cancer cells in a dose-dependent manner with the IC50 value of 2.41 ± 0.29 and 3.51 ± 0.14, respectively. The cytotoxic effect of the compound was further confirmed with a dose-dependent increase in the number of dead cells and enhanced LDH level in the culture medium. This compound exerted its anti-proliferative effect by G2/M phase cell growth arrest in MDA-MB-231 breast cancer cells and induced apoptosis-mediated cell death, which involved characteristic changes in cell and nuclear morphology, phosphatidylserine externalization, mitochondrial membrane depolarization, and increased ROS level. Neither hepatotoxicity nor nephrotoxicity was detected in the biochemical and histopathological analysis confirming the safety characterization of this compound usage. Therefore, the results significantly confirmed the potential anticancer activity of a novel β-diiminato compound, as evidenced by the induction of cell cycle arrest and apoptosis, which might be driven by the ROS‑mediated mitochondrial death pathway. This compound can be a promising candidate for future anticancer drug design and TNBC treatment, and further preclinical and clinical studies are warranted.

Octadecaneuropeptide, ODN, Promotes Cell Survival against 6-OHDA-Induced Oxidative Stress and Apoptosis by Modulating the Expression of miR-34b, miR-29a, and miR-21in Cultured Astrocytes.

Astrocytes specifically synthesize and release endozepines, a family of regulatory peptides including octadecaneuropeptide (ODN). We have previously reported that ODN rescues neurons and astrocytes from 6-OHDA-induced oxidative stress and cell death. The purpose of this study was to examine the potential implication of miR-34b, miR-29a, and miR-21 in the protective activity of ODN on 6-OHDA-induced oxidative stress and cell death in cultured rat astrocytes. Flow cytometry analysis showed that 6-OHDA increased the number of early apoptotic and apoptotic dead cells while treatment with the subnanomolar dose of ODN significantly reduced the number of apoptotic cells induced by 6-OHDA. 6-OHDA-treated astrocytes exhibited the over-expression of miR-21 (+118%) associated with a knockdown of miR-34b (-61%) and miR-29a (-49%). Co-treatment of astrocytes with ODN blocked the 6-OHDA-stimulated production of ROS and NO and stimulation of Bax and caspase-3 gene transcription. Concomitantly, ODN down-regulated the expression of miR-34b and miR-29a and rescued the 6-OHDA-associated reduced expression of miR21, indicating that ODN regulates their expression during cell death. Transfection with miR-21-3p inhibitor prevented the effect of 6-OHDA against cell death. In conclusion, our study indicated that (i) the expression of miRNAs miR-34b, miR-29a, and miR-21 is modified in astrocytes under 6-OHDA injury and (ii) that ODN prevents this deregulation to induce its neuroprotective action. The present study identified miR-21 as an emerging candidate and as a promising pharmacological target that opens new neuroprotective therapeutic strategies in neurodegenerative diseases, especially in Parkinson's disease.

Apomorphine Suppresses the Progression of Steatohepatitis by Inhibiting Ferroptosis.

The role of ferroptosis in steatohepatitis development is largely unknown. We investigated (1) whether hepatocyte ferroptosis occurs in a gene-modified steatohepatitis model without modifying dietary components, (2) whether ferroptosis occurs at an early stage of steatohepatitis, and (3) whether apomorphine, recently reported as a ferroptosis inhibitor, can ameliorate steatohepatitis. Hepatocyte-specific PTEN KO mice were used. Huh 7 and primary cultured hepatocytes isolated from the mice were used in this study. The number of dead cells increased in 10-week-old PTEN KO mice. This cell death was suppressed by the administration of ferroptosis inhibitor ferrostatin-1 for 2 weeks. Apomorphine also ameliorated the severity of steatohepatitis. Treatment with ferroptosis inhibitors, including apomorphine, decreases the level of lipid peroxidase. Apomorphine suppressed cell death induced by RSL-3 (a ferroptosis inducer), which was not suppressed by apoptosis or necroptosis inhibitors. Apomorphine showed a radical trapping capacity with much more potent activity than ferrostatin-1 and Trolox, a soluble form of vitamin E. In addition, apomorphine activated nrf2 and its downstream genes, including HO-1 and xCT. In conclusion, ferroptosis occurs in steatohepatitis from an early stage in PTEN KO mice. In addition, apomorphine ameliorates the severity of steatohepatitis by inhibiting ferroptosis.

Toxic effect of mussel Mytilus galloprovincialis exposed to Ag–TiO2 and ZnTi2O4–TiO2 bicomponent nanoparticles

Nanoparticles (NPs) are widely used in various fields, including antifouling paints for ships and industrial structures submerged in water. The potential impact of NPs on aquatic organisms, particularly their potential toxicity, is a significant concern, as their negative impact has been relatively poorly studied. In this study, we evaluated the effect of different concentrations of bimetallic Ag–TiO₂ and ZnTi₂O₄-TiO₂ NPs, which could potentially be used in antifouling coatings, on the hemocytes of the Mediterranean mussel Mytilus galloprovincialis. Hemocytes were exposed to NPs at concentrations of 0.1–1 mg/L for 1 and 2 h, and the production of reactive oxygen species (ROS), levels of DNA damage, and number of dead cells were measured. Exposure to Ag–TiO₂ NPs at 1 mg/L concentration for 1 h suppressed ROS production in hemocytes and reduced the relative number of agranulocytes in cell suspensions, without inducing DNA damage or cell death. Exposure to ZnTi2O4–TiO2 NPs did not cause changes in the ratio of granulocytes to agranulocytes in suspensions, nor did it affect other functional parameters of hemocytes. However, after a 2 h exposure period, ZnTi2O4–TiO2 NPs (1 mg/L) significantly reduced the production of ROS by hemocytes. These findings suggest that Ag–TiO2 and ZnTi2O4–TiO2 NPs have low acute toxicity for marine bivalves.

Incarvine C and its analogues inhibit the formation of cell cytoskeleton by targeting Rac1

Ras-related C3 botulinum toxin substrate 1 (Rac1) has emerged as a key regulator in the treatment of cancer metastasis because of its involvement in the formation of cell plate pseudopods and effects on cell migration. In this study, we found that incarvine C, a natural product isolated from Incarvillea sinensis, and its seven analogues exhibited antitumour activity by inhibiting cell cytoskeleton formation, with moderate cytotoxicity. Accordingly, these compounds inhibited the cytoskeleton-mediated migration and invasion of MDA-MB-231 cells, with inhibition rates ranging from 37.30 % to 69.72 % and 51.27 % to 70.90 % in vitro, respectively. Moreover, they induced G2/M phase cell cycle arrest in MDA-MB-231 cells. A pull-down assay revealed that the interaction between Rac1 and its downstream effector protein PAK1 was inhibited by these compounds and that the compound Ano-6 exhibited substantial activity, with an inhibition rate of more than 90 %. Molecular docking showed that incarvine C and its analogues could bind to the nucleotide-binding pocket of Rac1, maintaining high levels of inactivated Rac1. As Ano-6 exhibited significant activity in vitro, its anti-cancer activity was tested in vivo. Four weeks of oral treatment with Ano-6 was well-tolerated in mice, and it induced a potential anti-tumour response in xenografts of MDA-MB-231 cells. Further studies demonstrated that Ano-6 was enriched in tumour tissues after 2 h of administration and induced an increase in the number of dead tumour cells. In summary, these findings not only reveal the mechanism of incarvine C but also provide a new molecular template for Rac1 inhibitors and identify a promising candidate for breast cancer treatment.

Preparation and Evaluation of Herbo- Mineral Scrub using Banana Peel Powder

The objective of this study to prepare good and e effctive Herbo-Mineral scrub for face.This research article includes formulation and evaluation of a face scrub for lightening using banana peel powder.our face skin comes in direct contact of environmental factors like pollutant, dust particles etc. and having number of dead cells .to remove the dead skin and make face skin lightening, healthy, cleaned some facial prepration required . This scrub prepration contain banana peel powder which are safer for use it also they possess antioxidant, anti-ageing, lightening, and emollient properties , this scrub is prepared using simple mixing method . The scrub contain the vitamins, minerals, nutrients for the skin. The prepared scrub was evaluated for evaluating various parameters including physical appearance, colour, texture, odour, pH, viscosity, irritability, washability, homogenecity, extrudability, and spreadability, successful outcomes were achieved across all tested parameters and uatedd fruitful results for all the parameter tested. Thus the prepared formulation can be used effectively as it shows good scrubbing properties and it can be used to make a healthy, clean, lightening and glowing skin,

Caki-1 Spheroids as a 3D Model for Studying Free Fatty Acid-Induced Proximal Tubule Lipotoxicity

Three-dimensional (3D) multicellular spheroid shows a biological complexity resembling native tissue in comparison to conventional monolayer cultures, making it a better model system to assess drug effcacy and cytotoxicity. The main aim of this study was to examine the usefulness of Caki-1, a human proximal tubule (PT) representative cell line, as a 3D model system for studying free fatty acid-induced PT lipotoxicity. Caki-1 spheroids were generated and maintained on ultra-low attachment plates and characterized regarding time-dependent changes in morphology and functionality. In optimal 3D culture conditions, Caki-1 cells formed well-defined large compact 3D spheroids with uniform morphology, good circularity, and increased diameter from days 2-10. There was no change when the spheroids were incubated with lipid-free bovine serum albumin (BSA, 0.5%) from day 6 to 10. Chronic exposure to palmitate (300 μM) resulted in dispersed and flattened spheroid morphology, with a larger number of dead cells in the peripheral layers and decreased spheroid core. Moreover, palmitate-treated spheroids showed a significant increase in caspase-3 activity at the external layers of the spheroids, suggesting that the detached cells underwent apoptosis. Simultaneous addition of oleate (200 μM) prevents palmitate-induced spheroid disintegration and cell death in Caki-1 3D culture. Mechanistically, palmitate triggered endoplasmic reticulum (ER) stress and autophagy dysfunction, as evidenced by significantly increased protein levels of ER stress markers AFT4 and CHOP, and autophagy substrate p62. Oleate upregulated adipose differentiation-related protein (ADRP, a lipid droplet-associated protein), and promoted lipid droplet formation in palmitate-treated Caki-1 cells. These results indicate that Caki-1 3D spheroids are a simple and reproducible in vitro system for lipotoxicity study. It can potentially be used for evaluating human renal proximal tubular toxicity and drug screening. NIH/NIGMS R16GM149499, NIH TL1DK136047. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.