Search

Given the lack of advances in Oral Squamous Cell Carcinoma (OSCC) therapy in recent years, pharmacological strategies to block OSCC-related signaling pathways have gained prominence. The present study aimed to evaluate the therapeutic potential of Arsenic Trioxide (ATO) concerning its antitumoral effects and the inhibition of the Hedgehog (HH) pathway in OSCC. Initially, ATO cytotoxicity was assessed in a panel of cell lines. Cell viability, cell cycle, death patterns, and cell morphology were analyzed, as well as the effect of ATO on the expression of HH pathway components. After the cytotoxic assay, HSC3 cells were chosen for all in vitro assays. ATO increased apoptotic cell death and nuclear fragmentation in the sub-G1 cell cycle phase and promoted changes in cell morphology. In addition, the reduced expression of GLI1 indicated that ATO inhibits HH activity. The present study provides evidence of ATO as an effective cytotoxic drug for oral cancer treatment in vitro. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is the most common type of liver cancer and the fourth cause of cancer-related deaths worldwide. Presently, a few drugs are available for HCC treatment and prevention, including both natural and synthetic compounds. In this study, a new chalcone, (E)-1-(2,4,6-triethoxyphenyl)-3-(3,4,5-trimethoxyphenyl)prop-2-en-1-one (ETTC), was synthesized and its effects and mechanisms of action over human hepatoma cells were investigated. Cytotoxic activity was revealed in HCC cells, while no effects were observed in normal hepatocytes. In HCC cells, ETTC caused subG1 cell cycle arrest and apoptosis, characterized by nuclear fragmentation. The activation of caspases 3/7 and 9, the increase in pro-apoptotic BAX, and the decrease in anti-apoptotic BCL-2 suggest the activation of the intrinsic pathway of apoptosis. ETTC mitochondrial targeting is confirmed by the reduction in mitochondrial membrane potential and Complex I activity together with levels of superoxide anion increasing. Our outcomes prove the potential mitochondria-mediated antitumor effect of newly synthesized chalcone ETTC in HCC. [ABSTRACT FROM AUTHOR]

[Display omitted] • Small molecule-based design and synthesis of carboxamide-appended pyrazole conjugates. • 5g exhibited multifaceted potential (nuclear fragmentation, upregulation of caspase 3 and 9 etc.) against MDA-MB-231 cells. • The synergistic combination of paclitaxel and 5g has demonstrated improved efficacy by several folds. • The non-toxic nature of 5g was established in zebrafish embryos. We report the design, synthesis, and validation of carboxamide-based pyrazole and isoxazole conjugates with a multifaceted activity against Breast Cancer Cell Line MDA-MB-231. The study established that amongst the series, N -(3,5-bis(trifluoromethyl)benzyl)-3-(3,4,5-trimethoxyphenyl)-1 H -pyrazole-5-carboxamide (5 g) exhibits the highest potency in inhibiting Breast Cancer Cell Line MDA-MB-231 with an IC 50 value of 15.08 ± 0.04 µM. The MDA‐MB‐231 cells, upon treatment with compound 5 g , exhibited characteristic apoptotic specific activities such as nuclear fragmentation, phosphatidylserine translocation to the outer plasma membrane, release of lactate dehydrogenase (LDH), and upregulation of caspase 3 and caspase 9 activities. Also, the modulation of pro and antiapoptotic proteins in 5 g treated MDA-MB-231 cells was revealed by membrane array analysis. More importantly, the combination of paclitaxel and compound 5 g has exhibited improved activity by several folds via their synergistic effects. [ABSTRACT FROM AUTHOR]

Modern medicine must shift its paradigm from conventional treatment strategies to theranostics to meet individuals' needs. Carbon dots, as a class of fluorescent materials, provide biocompatible and multifunctional solutions for a wide range of applications including clinical sensing, imaging, and drug delivery. Various studies have focused on their synthesis, photophysical properties, and innoxious nature but their potential biomedical applications for the in-vivo toxicity are yet limited. In this work, previously synthesized C-Dots (CDs) were utilized to determine their effect on the development of Drosophila melanogaster. Simultaneously, the genotoxic potential of CDs was evaluated on specific larval cell types that play important roles in immunological defence as well as growth and development. The gut organ toxicity of both CDs was studied using DAPI and DCFH-DA dyes wherein RS-CDs didn't show significant toxicity to the concentration 500 μg/mL whereas RS-Cys-CDs showed nuclear fragmentation and modest ROS (reactive oxygen species) production. Subsequently, trypan-blue assay, larvae crawling assay, touch sensitivity, adult phenotype, and survivability assay were performed. The trypan-blue assay shows the non-toxic nature of both CDs even at the concentration of 500 μg/mL. The high concentrations of RS-Cys-CD (500 μg/mL) were further associated with the alteration in touch behaviour and decrease in pupa hatching. In-vivo and ex-vivo fluorescence assessment of both the CDs exhibit bright fluorescence in green and red channels upon excitation at 485 and 577 nm respectively. The prominent imaging results from RS-Cys-CDs highlight the positive impact of surface modification. [Display omitted] • RS-CDs and RS-Cys-CDs are synthesized using Ruellia simplex. • Both CDs were physicochemically characterized previously. • Both CDs were tested on Drosophila melanogaster for in-vivo toxicity. • RS-CD were non-toxic whereas higher concentration of RS-Cys-CD show modest toxicity and showed potential bioimaging. [ABSTRACT FROM AUTHOR]

Single agent therapy in the treatment of cancer did not provide satisfactory clinical outcome in a long term due to drug resistance, cumulative toxicity and suboptimal efficacy. Therefore, combination therapies are apt to improve the quality of life in patients. In the present study, combinatorial effect of EPA and/or DHA with doxorubicin (Dox) was evaluated for their potential synergistic effect on human lung adenocarcinoma (A549) cells. The respective IC 50 values obtained for EPA, DHA and Dox were 170, 140 and 2.5 μmol/L and thus EPA and DHA were separately combined with Dox at constant ratio of 68:1 and 56:1 respectively, depending on the ratio of their IC 50 values. Different combinations of drugs revealed dose-dependent inhibition of cell viability and IC 50 values were reduced to nearly 3-fold (58.75 μmol/L) and 4.5-fold (31.5 μmol/L) in EPA-Dox and DHA-Dox combination respectively. Combination index (CI) revealed strong synergism (CI < 0.5) in DHA-Dox (0.49) and moderate synergism (CI > 0.5) in EPA-Dox (0.79) at 0.5 Fa (Fraction affected), which is further confirmed by dose reduction index and isobologram. The mechanisms underlying synergistic interactions involve intracellular ROS generation and MMP depolarisation which causes changes in the cell morphology viz. Nuclear fragmentation/condensation, membrane blebbing and early/late apoptotic cells. Moreover, cell cycle arrest was observed at the S and G2/M phase, with more prominent effect in G2/M phase. No toxicity was imposed on non-cancerous HEK-293 T cells. These findings suggest that EPA-Dox and DHA-Dox could be explored further for animal studies and clinical trials for lung adenocarcinoma treatment. [Display omitted] [ABSTRACT FROM AUTHOR]

Breast cancer is one of the most common cancers globally and a leading cause of cancer-related deaths among women. Despite the combination of chemotherapy with targeted therapy, including monoclonal antibodies and kinase inhibitors, drug resistance and treatment failure remain a common occurrence. Copper, complexed to various organic ligands, has gained attention as potential chemotherapeutic agents due to its perceived decreased toxicity to normal cells. The cytotoxic efficacy and the mechanism of cell death of an 8-aminoquinoline-naphthyl copper complex (Cu8AqN) in MCF-7 and MDA-MB-231 breast cancer cell lines was investigated. The complex inhibited the growth of MCF-7 and MDA-MB-231 cells with IC 50 values of 2.54 ± 0.69 μM and 3.31 ± 0.06 μM, respectively. Nuclear fragmentation, annexin V binding, and increased caspase-3/7 activity indicated apoptotic cell death. The loss of mitochondrial membrane potential, an increase in caspase-9 activity, the absence of active caspase-8 and a decrease of tumour necrosis factor receptor 1(TNFR1) expression supported activation of the intrinsic apoptotic pathway. Increased ROS formation and increased expression of haem oxygenase-1 (HMOX-1) indicated activation of cellular stress pathways. Expression of p21 protein in the nuclei was increased indicating cell cycle arrest, whilst the expression of inhibitor of apoptosis proteins (IAPs); cIAP1, XIAP and survivin were decreased, creating a pro-apoptotic environment. Phosphorylated p53 species; phospho-p53(S15), phospho-p53(S46), and phospho-p53(S392) accumulated in MCF-7 cells indicating the potential of Cu8AqN to restore p53 function in the cells. In combination, the data indicates that Cu8AqN is a useful lead molecule worthy of further exploration as a potential anti-cancer drug. [ABSTRACT FROM AUTHOR]

• The study aims at exploring the potential bioactive compound from the extract of Northeastern plant Garcinia cowa belonging to Clusiaceae family having leishmanicidal activity against Leishmania donovani. • HR-LC-MS based metabolomic profiling study revealed presence of characteristic phytoconstituents. • Utilizing computational tools, modern day technique of molecular docking used for virtual screening of α-mangostin and β-mangostin for ideal lead molecule identification. • The xanthoid, α-mangostin was observed to have inhibitory activity on the viability of the promastigotes, axenic amastigotes as well as intracellular amastigotes. • The compound induces oxidative and nitrosative burst in the parasite resulting in onset of inflammatory response led by overproduction of nitric oxide. • The oxidative stress further induces cellular damage in form of nuclear fragmentation and mitochondrial dysfunction ultimately leading to apoptotic cell death of parasite, substantiating the anti-leishmanial effect of α-mangostin. Owing to the persistent number of parasitic deaths, Visceral leishmaniasis continues to haunt several economically weaker sections of India. The disease causes over 30,000 deaths and threatens millions annually on a global scale. The standard pentavalent antimonials, on the other hand, are associated with health adversities and disease relapse. The current study is focused on the search for the most potential natural bioactive phytocompound from the bark extract of the Northeastern Indian plant, Garcinia cowa , that shows potent anti-leishmanial properties. The High Resonance Liquid Chromatography followed by Mass Spectrometry (HR-LCMS) study followed by an in silico molecular docking using computational tools revealed that α-mangostin might potentially possess antiparasitic activity. To validate the anti-leishmanial efficacy of the compound, a cell viability assay was performed, which demonstrated the parasite-specific inhibitory activity of α-mangostin; with IC 50 values ranging from 4.95 - 7.37 µM against the different forms of Leishmania donovani parasite. The flow cytometric analysis of the phytocompound treated parasites indicated an oxidative and nitrosative stress-mediated apoptotic cell death in the parasites, by the suggestive surge in nuclear fragmentation and mitochondrial dysfunction. Simultaneously, a cytokine profiling study suggested approximate two-to-three-fold upregulated levels of pro-inflammatory cytokines post-compound treatment, which is predicted to actively contribute to parasite-killing. α-mangostin was also found to reduce the chances of parasite survival by inhibiting arginase enzyme activity, which in favorable conditions facilitates its sustenance. This study thereby substantiates that α-mangostin significantly possesses anti-leishmanial potentiality that can be developed into a cure for this infectious disease. [Display omitted] [ABSTRACT FROM AUTHOR]

An algorithm of pre-equilibrium clustering of spectator matter based on the construction of the minimum spanning tree (MST) is presented. The algorithm was implemented in the Abrasion-Ablation Monte Carlo for Colliders (AAMCC) model designed to study the characteristics of spectator matter in collisions of relativistic nuclei. Due to accounting for the pre-equilibrium clusters in modelling 208Pb–208Pb collisions at the LHC, the agreement of simulation results with experimental data on the average multiplicities of spectator nucleons was improved. The results of the AAMCC-MST were compared with experimental data on the interactions of 197Au nuclei in nuclear photoemulsion. Comparison of the yields of spectator nuclei calculated for 16O–16O collisions with the yields measured in interactions of 16O with light nuclei of photoemulsion made it possible to estimate the effect of MST-clustering in small nuclear systems.

Single-event ion imaging enables the direct reconstruction of the relative stopping power (RSP) information required for ion-beam therapy. Helium ions were recently hypothesized to be the optimal species for such technique. The purpose of this work is to investigate the effect of secondary fragments on the image quality of helium CT (HeCT) and to assess the performance of a prototype proton CT (pCT) scanner when operated with helium beams in Monte Carlo simulations and experiment. Experiments were conducted installing the U.S. pCT consortium prototype scanner at the Heidelberg Ion-Beam Therapy Center (HIT). Simulations were performed with the scanner using the TOPAS toolkit. HeCT images were reconstructed for a cylindrical water phantom, the CTP404 (sensitometry), and the CTP528 (line-pair) [Formula: see text] ® modules. To identify and remove individual events caused by fragmentation, the multistage energy detector of the scanner was adapted to function as a [Formula: see text] telescope. The use of the developed filter eliminated the otherwise arising ring artifacts in the HeCT reconstructed images. For the HeCT reconstructed images of a water phantom, the maximum RSP error was improved by almost a factor 8 with respect to unfiltered images in the simulation and a factor 10 in the experiment. Similarly, for the CTP404 module, the mean RSP accuracy improved by a factor 6 in both the simulation and the experiment when the filter was applied (mean relative error 0.40% in simulation, 0.45% in experiment). In the evaluation of the spatial resolution through the CTP528 module, the main effect of the filter was noise reduction. For both simulated and experimental images the spatial resolution was  ∼4 lp cm-1. In conclusion, the novel filter developed for secondary fragments proved to be effective in improving the visual quality and RSP accuracy of the reconstructed images. With the filter, the pCT scanner is capable of accurate HeCT imaging.

Porcine sapelovirus (PSV) is an emerging pathogen associated with symptoms of enteritis, pneumonia, polioencephalomyelitis and reproductive disorders in swine, resulting in significant economic losses. Although PSV is reported to trigger cell apoptosis, its specific molecular mechanism is unclear. In this research, the cell apoptosis induced by PSV infection and its underlying mechanisms were investigated. The morphologic features of apoptosis include nuclear condensation and fragmentation, were observed after PSV infection. The cell apoptosis was confirmed by analyzing the apoptotic rates, caspase activation, and PARP1 cleavage. Caspase inhibitors inhibited the PSVinduced intrinsic apoptosis pathway and reduced viral replication. Among the proteins encoded by PSV, 2A is an important factor in inducing the mitochondrial apoptotic pathway. The conserved residues H48, D91, and C164 related to protease activity in PSV 2A were crucial for 2A-induced apoptosis. In conclusion, our results provide insights into how PSV induces host cell apoptosis. [ABSTRACT FROM AUTHOR]

Zinc oxide nanomaterial is a potential material in the field of cancer therapy. In this study, zinc oxide nanospheres (ZnO-NS) were synthesized by Sol-gel method using yeast extract as a non-toxic bio-template and investigated their physicochemical properties through various techniques such as FTIR, XR, DLS, and TEM. Furthermore, free zinc ions released from the zinc oxide nanosphere suspended medium were evaluated by using the ICP-AS technique. Therefore, the cytotoxicity of ZnO nanospheres and released Zn ions on both HuH7 and Vero cells was studied using the MTT assay. The data demonstrated that the effectiveness of ZnO nanospheres on HuH7 was better than free Zn ions. Similarly, ZnO-Ns were significantly more toxic to HuH7 cell lines than Vero cells in a concentration-dependent manner. The cell cycle of ZnO-Ns against Huh7 and Vero cell lines was arrested at G2/M. Also, the apoptosis assay using Annexin-V/PI showed that apoptosis of HuH7 and Vero cell lines by ZnO nanospheres was concentration and time-dependent. Caspase 3 assay results showed that the apoptosis mechanism may be intrinsic and extrinsic pathways. The mechanism of apoptosis was determined by applying the RT-PCR technique. The results revealed significantly up-regulated Bax, P53, and Cytochrome C, while the Bcl2 results displayed significant down-regulation and the western blot data confirmed the RT-PCR data. There is oxidative stress of the ZnO nanospheres and free Zn+2 ions. Results indicated that the ZnO nanospheres and free Zn+2 ions induced oxidative stress through increasing reactive oxygen species (ROS) and lipid peroxidation. The morphology of the HuH7 cell line after exposure to ZnO nanospheres at different time intervals revealed the presence of the chromatin condensation of the nuclear periphery fragmentation. Interestingly, the appearance of canonical ultrastructure features of apoptotic morphology of Huh7, Furthermore, many vacuoles existed in the cytoplasm, the majority of which were lipid droplets, which were like foamy cells. Also, there are vesicles intact with membranes that are recognized as swollen mitochondria. [ABSTRACT FROM AUTHOR]

Betulinic acid (BA), a pentacyclic triterpenoid found in certain plant species, has been reported to have several health benefits including antioxidant and anti-apoptotic properties. However, the mechanism by which BA confers these properties is currently unknown. Saccharomyces cerevisiae, a budding yeast with a short life cycle and conserved cellular mechanism with high homology to humans, was used as a model for determining the role of BA in aging and programmed cell death (PCD). Treatment with hydrogen peroxide (H2O2) exhibited significantly increased (30–35%) survivability of antioxidant (sod1Δ, sod2Δ, cta1Δ, ctt1Δ, and tsa1Δ) and anti-apoptotic (pep4Δ and fis1Δ) mutant strains when cells were pretreated with BA (30 µM) as demonstrated in spot and CFU (Colony forming units) assays. Measurement of intracellular oxidation level using the ROS-specific dye H2DCF-DA showed that all tested BA-pretreated mutants exhibited decreased ROS than the control when exposed to H2O2. Similarly, when mutant strains were pretreated with BA and then exposed to H2O2, there was reduced lipid peroxidation as revealed by the reduced malondialdehyde content. Furthermore, BA-pretreated mutant cells showed significantly lower apoptotic activity by decreasing DNA/nuclear fragmentation and chromatin condensation under H2O2-induced stress as determined by DAPI and acridine orange/ethidium bromide staining. In addition, BA treatment also extended the life span of antioxidant and anti-apoptotic mutants by ∼10–25% by scavenging ROS and preventing apoptotic cell death. Our overall results suggest that BA extends the chronological life span of mutant strains lacking antioxidant and anti-apoptotic genes by lowering the impact of oxidative stress, ROS levels, and apoptotic activity. These properties of BA could be further explored for its use as a valuable nutraceutical. [ABSTRACT FROM AUTHOR]

Chemotherapy side effects are minimized, and therapeutic results are enhanced by combination therapy, which is extensively utilized in cancer therapy. MTT experiment showed that compared to the free drugs calreticulin (CRT) and gemcitabine (GEM), GEM-CRT@NPs significantly increased the cytotoxicity of endometrial cancer (Ishikawa cells and KLE cells). These drugs were conjugated to a tri-block copolymeric framework. GEM-CRT@NPs were studied using HR-TEM and DLS analysis to determine particle size and zeta potential. Biochemical staining analyses were also used to investigate the morphological characteristics of endometrial cancer cell lines. After KLE and Ishikawa cancer cells were treated with GEM-CRT@NPs, endometrial cell shape was altered, as evidenced by nuclear fragmentation and nuclear condensation (apoptosis hallmarks). Additionally, the most significant apoptosis ratio to mitochondrial membrane potential indicates that the mitochondrial membrane potential is responsible for the cell's death. Upon treatment with GEM-CRT@NPs, systemic toxicity analysis revealed minimal histological abnormalities and significant blood chemistry alterations, and a near-normal look of the organs. Endometrial cancer treatment may be enhanced with the use of the targeted combination therapeutic system. [ABSTRACT FROM AUTHOR]

Although apoptosis of podocytes has been widely reported in in vitro studies, it has been less frequently and less definitively documented in in vivo situations. To investigate this discrepancy, we analyzed the dying process of podocytes in vitro and in vivo using LMB2, a human (h)CD25-directed immunotoxin. LMB2 induced cell death within 2 days in 56.8 ± 13.6% of cultured podocytes expressing hCD25 in a caspase-3, Bak1, and Bax-dependent manner. LMB2 induced typical apoptotic features, including TUNEL staining and fragmented nuclei without lactate dehydrogenase leakage. In vivo, LMB2 effectively eliminated hCD25- expressing podocytes in NEP25 mice. Podocytes injured by LMB2 were occasionally stained for cleaved caspase-3 and cleaved lamin A but never for TUNEL. Urinary sediment contained TUNEL-positive podocytes. To examine the effect of glomerular filtration, we performed unilateral ureteral obstruction in NEP25 mice treated with LMB2 1 day before euthanasia. In the obstructed kidney, glomeruli contained significantly more cleaved lamin A-positive podocytes than those in the contralateral kidney (50.1 ± 5.4% vs. 29.3 ± 4.1%, P < 0.001). To further examine the dying process without glomerular filtration, we treated kidney organoids generated from nephron progenitor cells of NEP25 mice with LMB2. Podocytes showed TUNEL staining and nuclear fragmentation. These results indicate that on activation of apoptotic caspases, podocytes are detached and lost in the urine before nuclear fragmentation and that the physical force of glomerular filtration facilitates detachment. This phenomenon may be the reason why definitive apoptosis is not observed in podocytes in vivo. [ABSTRACT FROM AUTHOR]

The present study was carried out to investigate the effect of Arctigenin on cell growth and the mechanism of cell death elicited by Arctigenin were examined in FaDu human pharyngeal carcinoma cells. To determine the apoptotic activity of Arctigenin in FaDu human pharyngeal carcinoma cells, cell viability assay, DAPI staining, caspase activation analysis, and immunoblotting were performed. Arctigenin inhibited the growth of cells in a dose-dependent manner and induced nuclear condensation and fragmentation. Arctigenin-treated cells showed caspase-3/7 activation and increased apoptosis versus control cells. FasL, a death ligand associated with extrinsic apoptotic signaling pathways, was up-regulated by Arctigenin treatment. Moreover, caspase-8, a part of the extrinsic apoptotic pathway, was activated by Arctigenin treatments. Expressions of anti-apoptotic factors such as Bcl-2 and Bcl-xL, components of the mitochondria-dependent intrinsic apoptosis pathway, significantly decreased following Arctigenin treatment. The expressions of pro-apoptotic factors such as BAX, BAD and caspase-9, and tumor suppressor -53 increased by Arctigenin treatments. In addition, Arctigenin activated caspase-3 and poly (ADP-ribose) polymerase (PARP) induced cell death. Arctigenin also inhibited the proliferation of FaDu cells by the suppression of p38, NF-κB, and Akt signaling pathways. These results suggest that Arctigenin may inhibit cell proliferation and induce apoptotic cell death in FaDu human pharyngeal carcinoma cells through both the mitochondria-mediated intrinsic pathway and the death receptormediated extrinsic pathway. [ABSTRACT FROM AUTHOR]

Non-small cell lung cancer (NSCLC) is an important sub-type of lung cancer associated with poor diagnosis and therapy. Innovative multi-functional systems are urgently needed to overcome the invasiveness of NSCLC. Carbon quantum dots (CQDs) derived from natural sources have received interest for their potential in medical bio-imaging due to their unique properties, which are characterized by their water solubility, biocompatibility, simple synthesis, and low cytotoxicity. In the current study, ethylene-diamine doped CQDs enhanced their cytotoxicity (98 ± 0.4%, 97 ± 0.38%, 95.8 ± 0.15%, 86 ± 0.15%, 12.5 ± 0.14%) compared to CQDs alone (99 ± 0.2%, 98 ± 1.7%, 96 ± 0.8%, 93 ± 0.38%, 91 ± 1.3%) at serial concentrations (0.1, 1, 10, 100, 1000 μg/mL). In order to increase their location in a specific tumor site, folic acid was used to raise their functional folate recognition. The apoptotic feature of A549 lung cells exposed to N-CQDs and FA-NCQDs was characterized by a light orange-red color under fluorescence microscopy. Additionally, much nuclear fragmentation and condensation were seen. Flow cytometry results showed that the percentage of cells in late apoptosis and necrosis increased significantly in treated cells to (19.7 ± 0.03%), (27.6 ± 0.06%) compared to untreated cells (4.6 ± 0.02%), (3.5 ± 0.02%), respectively. Additionally, cell cycle arrest showed a strong reduction in cell numbers in the S phase (14 ± 0.9%) compared to untreated cells (29 ± 0.5%). Caspase-3 levels were increased significantly in A549 exposed to N-CQDs (2.67 ± 0.2 ng/mL) and FA-NCQDs (3.43 ± 0.05 ng/mL) compared to untreated cells (0.34 ± 0.04 ng/mL). The functionalization of CQDs derived from natural sources has proven their potential application to fight off non-small lung cancer. [ABSTRACT FROM AUTHOR]

This study evaluates protosappanin B's (PSB) role in inflammation, proliferation, and subsequent apoptosis of human melanoma A875 cells. The cell lines were treated with PSB (15 and 20 μM) for 24 h of incubation mediated cytotoxicity was assessed by MTT assay; ROS was studied by flow cytometry. PSB treatment mediated nuclear fragmentation and apoptotic morphological features were analyzed by AO, EtBr, PI, and DAPI stainings. The mRNA and protein expression of inflammatory, proliferation, cell survival and apoptosis were studied by RT-PCR and western blotting. PSB (15 and 20 μM) treatment with A875 produces cytotoxicity associated with nuclear fragmentation, and apoptotic changes were observed. PSB inhibits inflammatory proteins (TNF-α, NF-κB, COX-2, IL-6, iNOS, and VEGF) in A875 cells. Furthermore, PSB induces pro-apoptotic protein expressions i.e. Bax, caspase-3; inhibiting anti-apoptotic, cell proliferative proteins (cyclin-D1, Bcl-2, c-Myc, and survivin) in A875 cells. The p-PI3K, p-AKT, and p-GSK-3β contribute to oncogenic progression; hence, inhibition of these signaling events is considered a significant target for the treatment of melanoma. This study observed that PSB treatment magnificently inhibited the p-PI3K, p-AKT, and p-GSK-3β in A875 cells. Thus, PSB inhibits inflammation, proliferation, and survival; promotes apoptosis by suppressing PI3K, AKT, and GSK-3β in human melanoma cancer. [Display omitted] • Protosappanin-B concern as a natural drug that has numerous biological activity. • Protosappanin-B inhibits inflammation and proliferation in A875 cells. • Protosappanin-B inhibits melanoma cancer via suppression of PI3K/AKT expressions. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao is the property of Chinese Journal of Preventive Veterinary Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Exfoliation syndrome presents as an accumulation of insoluble fibrillar aggregates that commonly correlates with age and causes ocular complications, most notably open-angle glaucoma. Despite advances in understanding the pathogenesis and risk factors associated with exfoliation syndrome, there has been no significant progress in curative pharmacotherapy of this disease. It is thought that the ability to target the fibrillar aggregates associated with exfoliation may offer a new therapeutic approach, facilitating their direct removal from affected tissues. Phage display techniques yielded two peptides (LPSYNLHPHVPP, IPLLNPGSMQLS) that could differentiate between exfoliative and non-affected regions of the human lens capsule. These peptides were conjugated to magnetic particles using click chemistry to investigate their ability in targeting and removing exfoliation materials from the anterior human lens capsule. The behavior of the fibrillar materials upon binding to these magnetic particles was assessed using magnetic pins and rotating magnetic fields of various strengths. Ex vivo studies showed that the magnetic particle-peptide conjugates could generate enough mechanical force to remove large aggregates of exfoliation materials from the lens capsule when exposed to a low-frequency rotating magnetic field (5000 G, 20 Hz). Biocompatibility of targeting peptides with and without conjugated magnetic particles was confirmed using MTT cell toxicity assay, live/dead cell viability assay, and DNA fragmentation studies on primary cultured human trabecular meshwork cells. This is a novel, minimally invasive, therapeutic approach for the treatment of exfoliation glaucoma via the targeting and removal of exfoliation materials that could be applied to all tissues within the anterior segment of the eye. [ABSTRACT FROM AUTHOR]

We report the discovery of a new abscisic acid (ABA) metabolite, found in the course of a mass spectrometric study of ABA metabolism by the rhizosphere bacterium Rhodococcus sp. P1Y. Analogue of (+)-ABA, enriched in tritium in the cyclohexene moiety, was fed in bacterial cells, and extracts containing radioactive metabolites were purified and analyzed to determine their structure. We obtained mass spectral fragmentation patterns and nuclear magnetic resonance spectra of a new metabolite of ABA identified as 1-hydroxy-2,6,6-trimethyl-4-oxo-2-cyclohexene-1-acetic acid, which we named rhodococcal acid (RA) and characterized using several other techniques. This metabolite is the second bacterial ABA degradation product in addition to dehydrovomifoliol that we described earlier. Taken together, these data reveal an unknown ABA catabolic pathway that begins with side chain disassembly, as opposed to the conversion of the cyclohexene moiety in plants. The role of ABA-utilizing bacteria in interactions with other microorganisms and plants is also discussed. [ABSTRACT FROM AUTHOR]

Cucurbitacin-B (Cur-B) is an analogue triterpenoid belonging to the Cucurbitaceae family. Previous reports have explicitly outlined various biological activities of Cucurbitaceae family members, including the anticancer activity of Cur-B. In the present study, we tried to elucidate the anticancer efficacy of Cur-B against prostate cancer PC3 cells. PC3 cells were exposed to purified Cur-B at 5, 10, 15, 20 and 25 µM for 24. Cur-B exposure reduced cell viability of PC3 cells at 5 µM (p < 0.05), with further reduction with increased Cur-B concentration (15 µM, p < 0.01 and 25 µM, p < 0.001). Cur-B also succeeded in instigating nuclear fragmentation and condensation, followed by activation of caspase-8, -9 and -3 proportionally with increasing concentrations of Cur-B. Treatment with Cur-B also instigated ROS-mediated oxidative stress both qualitatively and quantitatively at 5 µM, p < 0.05; 15 µM, p < 0.01 and 25 µM, p < 0.001. Increased ROS after Cur-B treatment also led to dissipation of mitochondrial membrane potential, thereby resulting in considerable apoptosis (p < 0.001), which, again, was proportionally dependent on Cur-B concentration. Cur-B exposure to PC3 cells was concomitantly followed by reduced cyclin D1, cyclin-dependent kinase 4 (CDK4) expression and augmented mRNA expression of CDK inhibitor p21Cip1. Intriguingly, Cur-B exposure also led to considerable downregulation of the JAK/STAT signaling cascade, which may be the reason behind Cur-B-mediated apoptosis and cell cycle arrest within PC3 cells. Therefore, these observations explicitly establish that Cur-B could serve in the prevention of prostate cancer. [ABSTRACT FROM AUTHOR]

We investigate the contributions of various reaction channels to the interaction, reaction, charge-changing and neutron-changing cross sections. The goal is to investigate the relation between microscopic interactions and the symmetry energy component of the equation of state (EoS) of interest for the structure of neutron stars. We compare the neutron skins extracted from diverse experimental techniques with those obtained with Hartree–Fock–Bogoliubov calculations using 23 Skyrme and with eight density-dependent interactions used in the relativistic mean-field method. We show that no particular conclusion can be drawn on the best EoS in view of the wide range of uncertainty in the experimental data. We further investigate the prospects of using neutron-changing reactions to assess the isospin dependence of the neutron skin in neutron-rich nuclei. [ABSTRACT FROM AUTHOR]

Diindolylmethane (DIM) is a metabolic product of indole‐3‐carbinol (I3C), the major phytochemicals present in cruciferous vegetables, which can modulate multiple signalling pathways in cancer. The present study deals with the mechanism of action of two synthetic biaryl conjugates of DIM in triple negative breast cancer cells. Out of 12 DIM derivatives tested, two compounds, DIM‐1 and DIM‐4, exhibit cytotoxicity with GI50 values of 9.83 ± 0.2195 μM and 8.726 ± 0.5234 μM, respectively, in 2D culture. In 3D culture, DIM‐1 and DIM‐4 show GI50 values of 24.000 ± 0.7240 μM and 19.230 ± 0.3754 μM, respectively. The non‐toxic nature of the compounds was also established by the toxicity studies using the zebrafish model system. The two compounds induced apoptosis and anoikis in the cancer cells, which was confirmed by morphological analysis, nuclear fragmentation, membrane integrity assay, caspase activity measurements and modulation of pro/anti‐apoptotic proteins. The compounds inhibited cell migration and MMP‐2 and MMP‐9 activities indicating their anti‐metastatic property. They also reduced the expression of active Ras, phosphorylated forms of PI3K, Akt and mTOR. Immunofluorescence studies revealed the reduced expression of EGFR and pEGFR in treated cells. To conclude, DIM‐1 and DIM‐4 induced anti‐breast cancer effects by blocking EGF receptor and subsequently inhibiting Ras‐mediated PI3K‐Akt–mTOR signalling pathway. [ABSTRACT FROM AUTHOR]

Farnesiferol C (Far-C) is a coumarin commonly extracted from Ferula asafetida and is popularly used as a traditional source of natural remedy. Liver cancer or hepatocellular carcinoma (HCC) has emerged as a major cause behind cancer burden, and limited therapeutic interventions have further aggravated the clinical management of HCC. In the present study, the authors tested the hypothesis that Far-C-instigated oxidative stress resulted in anti-proliferation and apoptosis instigation within human liver cancer HepG2 cells. The observations reported herewith indicated that Far-C exerted considerable cytotoxic effects on HepG2 cells by reducing the cell viability (p < 0.001) in a dose-dependent manner. Far-C exposure also resulted in enhanced ROS production (p < 0.01) which subsequently led to loss of mitochondrial membrane potential. Far-C-instigated oxidative stress also led to enhanced nuclear fragmentation and condensation as revealed through Hoechst-33342. These molecular changes post-Far-C exposure also incited apoptotic cell death which concomitantly led to significant activation of caspase-3 (p < 0.001). Furthermore, Far-C exhibited its competence in altering the expression of genes involved in apoptosis regulation (Bax, Bad, and Bcl2) along with genes exerting regulatory effects on cell cycle (cyclinD1) and its progression (p21Cip1 and CDK4). The evidence thus clearly shows the preclinical efficacy of Far-C against HepG2 cells. However, further mechanistic investigations deciphering the alteration of different pathways post-Far-C exposure will be highly beneficial. [ABSTRACT FROM AUTHOR]

Mitochonic Acid 5 (MA-5) enhances mitochondrial ATP production, restores fibroblasts from mitochondrial disease patients and extends the lifespan of the disease model "Mitomouse". Additionally, MA-5 interacts with mitofilin and modulates the mitochondrial inner membrane organizing system (MINOS) in mammalian cultured cells. Here, we used the nematode Caenorhabditis elegans to investigate whether MA-5 improves the Duchenne muscular dystrophy (DMD) model. Firstly, we confirmed the efficient penetration of MA-5 in the mitochondria of C. elegans. MA-5 also alleviated symptoms such as movement decline, muscular tone, mitochondrial fragmentation and Ca2+ accumulation of the DMD model. To assess the effect of MA-5 on mitochondria perturbation, we employed a low concentration of rotenone with or without MA-5. MA-5 significantly suppressed rotenone-induced mitochondria reactive oxygen species (ROS) increase, mitochondrial network fragmentation and nuclear destruction in body wall muscles as well as endogenous ATP levels decline. In addition, MA-5 suppressed rotenone-induced degeneration of dopaminergic cephalic (CEP) neurons seen in the Parkinson's disease (PD) model. Furthermore, the application of MA-5 reduced mitochondrial swelling due to the immt-1 null mutation. These results indicate that MA-5 has broad mitochondrial homing and MINOS stabilizing activity in metazoans and may be a therapeutic agent for these by ameliorating mitochondrial dysfunction in DMD and PD. [ABSTRACT FROM AUTHOR]

In the present study, we aimed to confirm whether NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) could induce apoptosis of human hepatic stellate cells (HSCs) LX-2 and explore the potential pharmacological mechanism underlying these effects. In this study, LX-2 cells were cultured in vitro, and the experiment was divided into two groups, including the control and PDTC groups. The viability of LX-2 cells was measured by CCK8 assay after the cells were exposed to PDTC. The anti-apoptotic effect of PDTC was detected by AO/EB double assay staining kit. Additionally, the activities of NF-κB, Fas/FasL, apoptosis-related proteins, as well as the cellular localization of AIF, were determined by Western blotting analysis and immunofluorescence staining respectively. After PDTC treatment for 12 and 24 h, AO/EB dual staining showed typical apoptotic changes, such as cell volume reduction, cell shrinkage, nuclear fragmentation, and so on. PDTC at 60 μmol/L significantly increased the proliferation inhibition rate and decreased the secretion of collagen I, collagen III, and α-SMA in LX-2 cells. The Western blotting analysis and RT-PCR showed no significant difference in the expression of AIF between the control group and PDTC group, and the expressions of Fas and FasL were not observed in all groups (P > 0.05). Further results showed that PDTC could promote the displacement of AIF from mitochondria to the nucleus, activate the apoptotic signaling in the cell nucleus, and possibly participate in the apoptosis process of LX-2 cells. In conclusion, the pharmacological mechanism of PDTC against hepatic fibrosis might be to promote the displacement of AIF from mitochondria to the nucleus, then activate the apoptotic signaling in the cell nucleus, and finally induce the apoptosis of LX-2 cells. Meanwhile, these results also revealed that the Fas/FasL-mediated apoptosis pathway was not involved in the PDTC-induced apoptosis process of LX-2 cells. [ABSTRACT FROM AUTHOR]

The dendritic growth layer (DGL), defined as the temperature region between -20 and -10 ∘ C, plays an important role for ice depositional growth, aggregation and potentially secondary ice processes. The DGL has been found in the past to exhibit specific observational signatures in polarimetric and vertically pointing radar observations. However, consistent conclusions about their physical interpretation have often not been reached. In this study, we exploit a unique 3-months dataset of mid-latitude winter clouds observed with vertically pointing triple-frequency (X-, Ka-, W-band) and polarimetric W-band Doppler radars. In addition to standard radar moments, we also analyse the multi-wavelength and polarimetric Doppler spectra. New variables, such as the maximum of the spectral differential reflectivity (ZDR) (s ZDR max), allows us to analyse the ZDR signal of asymmetric ice particles independent of the presence of low ZDR producing aggregates. This unique dataset enables us to investigate correlations between enhanced aggregation and evolution of small ice particles in the DGL. For this, the multi-frequency observations are used to classify all profiles according to their maximum average aggregate size within the DGL. The strong correlation between aggregate class and specific differential phase shift (KDP) confirms the expected link between ice particle concentration and aggregation. Interestingly, no correlation between aggregation class and s ZDR max is visible. This indicates that aggregation is rather independent of the aspect ratio and density of ice crystals. A distinct reduction of mean Doppler velocity in the DGL is found to be strongest for cases with largest aggregate sizes. Analyses of spectral edge velocities suggest that the reduction is the combined result of the formation of new ice particles with low fall velocity and a weak updraft. It appears most likely that this updraft is the result of latent heat released by enhanced depositional growth. Clearly, the strongest correlations of aggregate class with other variables are found inside the DGL. Surprisingly, no correlation between aggregate class and concentration or aspect ratio of particles falling from above into the DGL could be found. Only a weak correlation between the mean particle size falling into the DGL and maximum aggregate size within the DGL is apparent. In addition to the correlation analysis, the dataset also allows study of the evolution of radar variables as a function of temperature. We find the ice particle concentration continuously increasing from -18 ∘ C towards the bottom of the DGL. Aggregation increases more rapidly from -15 ∘ C towards warmer temperatures. Surprisingly, KDP and s ZDR max are not reduced by the intensifying aggregation below - 15 ∘ C but rather reach their maximum values in the lower half of the DGL. Also below the DGL, KDP and s ZDR max remain enhanced until -4 ∘ C. Only there, additional aggregation appears to deplete ice crystals and therefore reduce KDP and s ZDR max. The simultaneous increase of aggregation and particle concentration inside the DGL necessitates a source mechanism for new ice crystals. As primary ice nucleation is expected to decrease towards warmer temperatures, secondary ice processes are a likely explanation for the increase in ice particle concentration. Previous laboratory experiments strongly point towards ice collisional fragmentation as a possible mechanism for new particle generation. The presence of an updraft in the temperature region of maximum depositional growth might also suggest an important positive feedback mechanism between ice microphysics and dynamics which might further enhance ice particle growth in the DGL. [ABSTRACT FROM AUTHOR]

Oyster mushrooms form an integral part of many diets owing to their characteristic aroma, delicious taste and nutraceutical value. In this study, we examined oyster mushrooms by direct arc optical emission spectroscopy for the presence of various biologically important elements. Furthermore, we screened phytochemicals present in Pleurotus ostreatus by applying GC-MS. Additionally, the antioxidant, antibacterial and anticancer activities of the ethanolic extract of Pleurotus ostreatus were studied. Moreover, we docked the phytochemicals and examined their binding affinities with EGFR, PR and NF-κB proteins, which are overexpressed in breast cancer. The elemental analysis showed the presence of Fe, K, Na, Ca, Mg, Cr and Sr in the spectrum. Moreover, GC-MS data revealed the presence of 32 biologically active compounds in oyster mushrooms. The ethanolic extract displayed remarkable free radical scavenging activity (~50%) against DPPH. The mushroom has shown promising antibacterial activity against both Gram-positive (S. aureus) and Gram-negative bacteria (Pseudomonasaeruginosa, Proteus vulgaris and Proteus mirabilis). The present study also revealed that oyster mushrooms possess significant anticancer activity. The ethanolic extract inhibited the growth and proliferation of MCF-7 cells. It also induced cell shrinkage, membrane blebbing and nuclear fragmentation, resulting in apoptosis of malignant cells. The molecular docking analysis showed that ligand 15 (Linoleic acid ethyl ester), ligand 27 (Ergosta-5,7,9(11),22-tetraen-3-ol, (3. beta.,22E), ligand 28 (Stigmasta-5,22-dien-3-ol, acetate, (3. beta.,22Z), ligand 30 (Ergosta-5,7,22-Trien-3-Ol, (3. Beta.,22E) and ligand 32 (gamma. Sitosterol) exhibited better binding affinities with EGFR, PR and NF-κB proteins. This result provides a strong ground for confirmation of the in vitro anticancer effect of Pleurotus ostreatus. From the present in vitro and in silico studies, it can be concluded that Pleurotus ostreatus is a useful source of essential elements and reservoir of bioactive compounds which confer its significant antioxidant, antibacterial and anticancer properties. [ABSTRACT FROM AUTHOR]

The number of grains per panicle significantly contributes to rice yield, but the regulatory mechanism remains largely unknown. Here, we reported a loss-of-function mutant, panicle apical abortion 7 (paa7), which exhibited panicle abortion and degeneration of spikelets on the apical panicles during the late stage of young panicle development in rice. High accumulations of H2O2 in paa7 caused programmed cell death (PCD) accompanied by nuclear DNA fragmentation in the apical spikelets. Map-based cloning revealed that the 3 bp "AGC" insertion and 4 bp "TCTC" deletion mutation of paa7 were located in the 3′-UTR regions of LOC_Os07g47330, which was confirmed through complementary assays and overexpressed lines. Interestingly, LOC_Os07g47330 is known as FRIZZY PANICLE (FZP). Thus, PAA7 could be a novel allele of FZP. Moreover, the severe damage for panicle phenotype in paa7/lax2 double mutant indicated that PAA7 could crosstalk with Lax Panicle 2 (LAX2). These findings suggest that PAA7 regulates the development of apical spikelets and interacts with LAX2 to regulate panicle development in rice. [ABSTRACT FROM AUTHOR]

Background: During the mushroom production process, about one-fifth of the mushroom gets lost. The mushroom residues (MR) are rich in nutrients and can be utilized in diverse applications. Therefore, the goal of this research was to modify a local hammermill to improve the performance of chopping mushroom residues to be efficient used as a by-product. Results: The experiments were conducted on a hammermill without and with six screen diameters, three drum rotational speeds, three feed rates, two average moisture content and two hammer rotation tracks (long–short). Then, chopper experiments were carried out with a focus on a specified size, power consumption and energy required. The findings of using any size screen offered little productivity since after a short time, the chopping MR was built blocks around the drum and blades. But using no screen gives these phenomena disappear. The chopping MR performed well in long track conditions, with a feeding rate of 700 kg/h, a drum speed of 300 rpm and moisture content of 43%. Conclusions: The chopping operation had the best at feed rate of 700 kg/h, drum speed of 3000 rpm and 43% moisture content of which gave the appropriate MWD of 5.54 and 5.32 mm, consumed power of 1114.35 and 1189.125 W and required specific energy of 1.59 and 1.7 kW h/Mg for short and long tracks, respectively, and also largest mean weight diameter under such conditions due to a decrease in the amount of mushroom residues and an increase in their dryness, which increases the fragmentation impact of hammers on mushroom remnants. [ABSTRACT FROM AUTHOR]

The process of spermatogenesis features significant germ cell loss through apoptosis. Routine histology of the testes of well-studied animal models hardly discloses any trace of their phagocytic clearance by the supporting Sertoli cells. This review highlights lessons learnt from the cystic, diametric testes of some seasonally migrating elasmobranchs (e.g., spiny dogfish and blue sharks) that offer unconventional investigative paradigms to study these phenomena as these organs readily disclose a pronounced apoptosis gradient affecting exclusively spermatogonial clones that each are enclosed with their own Sertoli cells in spherical structures called spermatocysts. This gradient is visible at a certain time of year in the spermatogenically active shark, and peaks in mature spermatogonial cysts as clustered deaths with sporadic, and not massive secondary necrosis. Conversely, immature spermatogonial cysts in blue sharks reveal a characteristic periluminal display of single apoptotic deaths. Tracing aberrations in the immunostaining patterns of the conserved cell cycle marker, proliferating cell nuclear antigen, the gradual progression of the death process in individual or coalesced spermatogonia in contiguous cysts becomes clear. The multiple apoptotic nuclear fragmentation morphologies inform also of a protracted death process involving three different morphological routes of nuclear fragmentation (of which some are TUNEL-positive and other TUNEL-negative) and concomitant chromatin compaction that culminate in freed apoptotic bodies (i.e., secondary necrosis). It is discussed that the staggered spermatogonial deaths and accompanying intermittent secondary necrosis in mature blue shark spermatogonial cysts may well relate to the low phagocytosis capacity of cyst's Sertoli cells that are still functionally naïve. [ABSTRACT FROM AUTHOR]

Podocytes are highly specialized cells playing a key role in the filtration function of the kidney. A damaged podocyte ultrastructure is associated with a reorganization of the actin cytoskeleton and accompanied with a loss of adhesion to the glomerular basement membrane leading to proteinuria in many forms of glomerular diseases, e.g. nephrotic syndrome. If the first-line therapy with glucocorticoids fails, alternative immunosuppressive agents are used, which are known to have the potential to stabilize the actin cytoskeleton. A new option for preventing relapses in steroid dependent nephrotic syndrome is the monoclonal antibody rituximab, which, in addition to its B-cell depleting effect, is assumed to have direct effects on podocytes. We here provide data on the non-immunological off-target effects of the immunosuppressant rituximab on podocyte structure and dynamics in an in vitro puromycin aminonucleoside model of podocyte injury. A conditionally immortalized human podocyte cell line was used. Differentiated podocytes were treated with puromycin aminonucleoside and rituximab. Our studies focussed on analyzing the structure of the actin cytoskeleton, cellular adhesion and apoptosis using immunofluorescence staining and protein biochemistry methods. Treatment with rituximab resulted in a stabilization of podocyte actin stress fibers in the puromycin aminonucleoside model, leading to an improvement in cell adhesion. A lower apoptosis rate was observed after parallel treatment with puromycin aminonucleoside and rituximab visualized by reduced nuclear fragmentation. Consistent with this data, Western-blot analyses demonstrated that rituximab directly affects the caspase pathways by inhibiting the activation of Caspases-8, -9 and -3, suggesting that rituximab may inhibit apoptosis. In conclusion, our results indicate an important role of the immunosuppressant rituximab in terms of stability and morphogenesis of podocytes, involving apoptosis pathways. This could help to improve therapeutical concepts for patients with proteinuria mediated by diseased podocytes. [ABSTRACT FROM AUTHOR]

Adenium obesum commonly known as "desert rose" belongs to the family Apopcynaceae and has previously been reported for its anti-influenza, antimicrobial, and cytotoxic efficacies and well-known for their ethno-medicinal applications. In the present study, ethanolic extracts of A. obesum (AOE) were analyzed by gas chromatography-mass spectrometry (GC-MS) to identify the important phytochemical compounds. The GC-MS analysis of AOE detected the presence of 26 phytochemical compounds. This plant is traditionally used for the treatment of various diseases. In this report, the antioxidant, antiinflammatory, and anticancer activities of ethanolic leaf extract from A. obesum (AOE) were studied. The antioxidant potential of ethanolic extract of AOE was examined by different antioxidant assays, such as antioxidant capacity by the DPPH, ABTS, superoxide, hydroxyl radical scavenging, and lipid peroxidation inhibition assays. The antioxidant activities of various reaction mixtures of AOE were compared with a reference or standard antioxidant (ascorbic acid). In addition, we also evaluated the anticancer activity of AOE, and it was observed that AOE was found to be cytotoxic against A549 lung cancer cells. It was found that AOE inhibited the viability of A549 lung cancer cells by inducing nuclear condensation and fragmentation. Furthermore, ethanolic AOE demonstrated the anti-inflammatory potential of AOE in murine alveolar macrophages (J774A.1) as an in vitro model system. AOE showed its potential in reducing the levels of inflammatory mediators including the proinflammatory cytokines and TNF-a. The results obtained in the present investigation established the antioxidant, anticancer, and anti-inflammatory potency of AOE, which may account for subsequent studies in the formulation of herbal-based medicine. [ABSTRACT FROM AUTHOR]

Due to having a long history of traditional uses as a functional food, Zingiber zerumbet was selected here to explore the inherent antioxidant and antineoplastic activities of methanolic extract of its rhizome (MEZZR) against Ehrlich ascites carcinoma (EAC) cells. The rich polyphenol containing MEZZR showed a marked DPPH, ABTS, nitric oxide radicals and lipid peroxidation inhibition activity with an IC50 of 3.43 ± 1.25, 11.38 ± 1.39, 23.12 ± 3.39 and 16.47 ± 1.47 µg/ml, respectively, when compared to the standard catechin. In vivo, MEZZR significantly inhibited EAC cell growth, decreased body weight gain, increased life span and restored the altered hematological characteristics of EAC-bearing mice. Moreover, MEZZR induced nuclear condensation and fragmentation, which are notable features of apoptosis as observed by fluorescence microscopy after staining EAC cells of MEZZR-treated mice with Hoechst 33342. Additionally, in vitro, the cell growth inhibition caused by the MEZZR in MTT assay, was remarkably decreased in the presence of caspase-3, -8 and -9 inhibitors. This study thus suggests that MEZZR may possess promising antiproliferative efficacy against EAC cells by inducing cell apoptosis. [ABSTRACT FROM AUTHOR]

Drug discovery is a challenging process with a huge molecular space to be explored and numerous pharmacological properties to be appropriately considered. Among various drug design protocols, fragment-based drug design is an effective way of constraining the search space and better utilizing biologically active compounds. Motivated by fragmentbased drug search for a given protein target and the emergence of artificial intelligence (AI) approaches in this field, this work advances the field of in silico drug design by (1) integrating a graph fragmentation-based deep generative model with a deep evolutionary learning process for large-scale multi-objective molecular optimization, and (2) applying protein-ligand binding affinity scores together with other desired physicochemical properties as objectives. Our experiments show that the proposed method can generate novel molecules with improved property values and binding affinities. [ABSTRACT FROM AUTHOR]

WRKY transcription factors (TFs) play crucial roles in biotic and abiotic stress responses. However, their roles in thermal response are still largely elusive, especially in rice. In this study, we revealed the functions of WRKY10 TF and VQ8 protein containing VQ motif in rice thermotolerance. Overexpression of WRKY10 or loss of VQ8 function increases thermosensitivity, whereas conversely, overexpression of VQ8 or loss of WRKY10 function enhances thermotolerance. Overexpression of WRKY10 accelerates reactive oxygen species (ROS) accumulation in chloroplasts and apoplasts, and it also induces the expression of heat shock TF and protein genes. We also found that WRKY10 regulates nuclear DNA fragmentation and hypersensitive response by modulating NAC4 TF expression. The balance between destructive and protective responses in WRKY10‐overexpression plant is more fragile and more easily broken by heat stress compared with wild type. In vitro and in vivo assays revealed that VQ8 interacts with WRKY10 and inhibits the transcription activity via repressing its DNA‐binding activity. Our study demonstrates that WRKY10 negatively regulates thermotolerance by modulating the ROS balance and the hypersensitive response and that VQ8 functions antagonistically to positively regulate thermotolerance. The functional module of WRKY10‐VQ8 provides safe and effective regulatory mechanisms in the heat stress response. Summary statement: The role of WRKY transcription factors in thermal response is largely elusive, especially in rice. Our results showed that WRKY10 and its interacting partner VQ8 played an antagonistic role, safely and effectively regulating rice thermotolerance by modulating reactive oxygen species (ROS) balance and hypersensitive response. [ABSTRACT FROM AUTHOR]

Aim: The capability of erythropoietin (EPO) as antioxidant is exported by the results of 2 preliminary studies, totally evaluating 4 histologic variables of endosalpingeal edema (EE) and oviductal congestion (OC) of the first one and endosalpingeal karyorrhexis (EK) and salpingitis (S) of the other one. Materials and Methods: The antioxidant capability was evaluated upon the 60th reperfusion min (for groups called A and C) and upon the 120th reperfusion min (for groups called B and D). The groups A and B were placebo ones, whereas the groups C and D included EPO as antioxidant. Results: The first pillar study showed that EPO has an oxidant non-significant potency for EE and OC together (p-values=0.5971); grade: "without lesions" alterations 0.0363636 [-0.1017439 - 0.1744712]. The second pillar study showed also an oxidant non-significant potency for EK and S; grade: "without lesions" alterations 0.0090909 [-0.0339659 - 0.0521478] (p-values=0.6715). The 2 above studies were added trying to calculate a common diagnostic value for all the four variables. Conclusion: EPO as antioxidant is a significant recessing agent for the total of the 4 histologic variables; grade: "without lesions" alterations score 0.0136364 [-0.0887489+0.0614762] (p-value=0.7153). [ABSTRACT FROM AUTHOR]

Background: Morinda elliptica (family Rubiaceae), locally known as 'mengkudu kecil', has been used by the Malays for medicinal purposes. Anthraquinones isolated from the roots of Morinda elliptica, namely nordamnacanthal and damnacanthal, have been widely reported to exhibit anticancer and antioxidant properties in various cancer models in vitro and in vivo. Aim: This study analyzed the morphological and ultrastructural effects of damnacanthal and nordamnacanthal on T-lymphoblastic leukemia CEM-SS cells. Method: Light microscopy, Giemsa staining, Wright's staining, scanning electron microscopy, and transmission electron microscopy were carried out to determine apoptosis, necrosis, and ultrastructural changes that occurred within the cells. Results: The outcomes showed that these compounds induced cell death by apoptosis and necrosis, specifically at higher doses of 10 and 30 μg/mL. Condensation and fragmentation of the nuclear chromatin, which further separated into small, membrane-bound vesicles known as apoptotic bodies, were observed in the nuclei and cytoplasm. The plasma membranes and cytoskeletons also showed marked morphological changes upon treatment with damnacanthal and nordamnacanthal, indicating apoptosis. Conclusion: Therefore, we report that damnacanthal and nordamnacanthal exhibit anticancer properties by inducing apoptosis and necrosis in CEM-SS cells, and they have potential as a drug for the treatment of T-lymphoblastic leukemia. [ABSTRACT FROM AUTHOR]

Nisin is an antibacterial peptide with anticancer properties, but the main drawback is its rapid enzymatic degradation and limited permeation across the cell membrane. This research aims to overcome these drawbacks by developing nisin-loaded nanoparticles (NPN) with improved cytotoxic effects. PLGA nanoparticles are one of the most effective biodegradable and biocompatible drug delivery carriers. In the present study, nisin-loaded nanoparticles showed enhanced anticancer effects. NPN was prepared by a double emulsion solvent evaporation method and characterized for different parameters. The cytotoxic investigation of NPN was carried out on various cell lines, including A549, SW-620, HT-29, PC-3, MDA-MB-231, MCF-7, MiaPaca-2, and fR2 by sulforhodamine B (SRB) assay. Mechanistic investigation of cellular cytotoxicity was performed by using bright-field microscopy, DAPI staining, intracellular reactive oxygen species (ROS), changes in mitochondrial membrane potential (ΔΨm), Western blotting and cellular uptake study. A comparative cytotoxicity study of nisin and NPN was performed on normal breast epithelial cells (fR2). NPN showed spherical shape, 289.09 ± 3.63 nm particle size, and 63.37 ± 3.12% entrapment efficiency. NPN was more cytotoxic to the MDA-MB-231 cell line, showing higher nuclear fragmentation, ROS generation, depletion of ΔΨm, and enhanced intracellular uptake with apoptosis signs compared with nisin and with no cytotoxicity on normal cells. The findings suggest that nisin delivery via PLGA nanoparticles can be used to treat cancer without significant effects on healthy cells. [ABSTRACT FROM AUTHOR]

Liposarcoma is a rare and heterogeneous soft tissue malignant tumor and has a significant impact on mortality with a poor prognosis. To date, there is no effective treatment for liposarcoma, whereas surgical resection is only the gold treatment with numerous adverse effects. Here we investigated whether tetrandrine inhibits the growth of SW872 human malignant liposarcoma cells. Of note, tetrandrine at 10 μM vastly inhibited growth and induced apoptosis, as evidenced by increased nuclear DNA fragmentation and sub-G1 population of SW872 cells. Mechanistically, treatment with tetrandrine led to activation of caspase-9/3 in SW872 cells, and z-VAD-fmk, a pan-caspase inhibitor, attenuated the tetrandrine-induced apoptosis and growth suppression in SW872 cells. In addition, tetrandrine treatment resulted in down-regulation of XIAP andSTAT-3 in SW872 cells, and importantly knockdown of STAT-3 caused a significant reduction of the cell survival. Tetrandrine also had abilities to up-regulate not only the expression of GRP78 and ATF-4 but also the phosphorylation of eIF-2α in SW872 cells. In summary, these results demonstrated that tetrandrine has strong growth-suppressive and apoptosis-inducing effects on SW872 cells, which are mediated through control of the intrinsic caspase pathway, down-regulation of XIAP and STAT-3, and triggering ER stress. [ABSTRACT FROM AUTHOR]

A class of plant defense and storage proteins, including Putranjiva roxburghii PNP protein (PRpnp), belongs to PNP-UDP family. The PRpnp and related plant proteins contain a disrupted PNP-UDP domain as revealed in previous studies. In PRpnp, the insert disrupting the domain contains the trypsin inhibitory site. In the present work, we analyzed native PRpnp (nPRpnp) complex formation with trypsin and inosine using SAXS experiments and established its dual functionality. Results indicated a relatively compact nPRpnp:Inosine structure, whereas trypsin complex showed conformational changes/flexibility. nPRpnp also exhibited a strong anti-cancer activity toward breast cancer (MCF-7), prostate cancer (DU-145) and hepatocellular carcinoma (HepG2) cell lines. MCF-7 and DU-145 were more sensitive to nPRpnp treatment as compared to HepG2. However, nPRpnp treatment showed no effect on the viability of HEK293 cells indicating that nPRpnp is specific for targeting the viability of only cancer cells. Further, acridine orange, DAPI and DNA fragmentation studies showed that cytotoxic effect of nPRpnp is mediated through induction of apoptosis as evident from the apoptosis-associated morphological changes and nuclear fragmentation observed after PRpnp treatment of cancer cells. These results suggest that PRpnp has the potential to be used as an anticancer agent. This is first report of anticancer activity as well as SAXS-based analysis for a PNP enzyme with trypsin inhibitory activity. [ABSTRACT FROM AUTHOR]

Overexpression of casein kinase 2 (CK2) has an oncogenic and pro-survival role in many cancers. CX-4945 (Silmitasertib) is a CK2 inhibitor with anti-cancerous and anti-angiogenic effects. Up to date, the anti-cancer effect and mechanism of CX-4945 on human cholangiocarcinoma (CCA) remain unclear. This study investigated whether CX-4945 inhibits growth and induces apoptosis of HuCCT-1 cells, a human CCA cell line. Of note, treatment with CX-4945 at 20 μM markedly reduced survival and induced apoptosis of HuCCT-1 cells, as evidenced by nuclear DNA fragmentation, PARP cleavage, activation of caspase-9/3, and up-regulation of DR-4. Although CX-4945 did not affect the phosphorylation and expression of CK2, it vastly inhibited the phosphorylation of CK2 substrates, supporting the drug's efficacy in inhibiting CK2 and its downstream pathway. Importantly, knockdown of CK2 that partially suppressed the phosphorylation of CK2 substrates resulted in a significant reduction of HuCCT-1 cell survival. In addition, CX-4945 reduced the phosphorylation and expression of STAT-3 and STAT-5 in HuCCT-1 cells, and pharmacological inhibition or respective knockdown of these proteins resulted in significant growth suppression of HuCCT-1 cells. CX-4945 also had abilities to decrease Mcl-1 expression while increasing eIF-2α phosphorylation in HuCCT-1 cells. Furthermore, there was a time-differential negative regulation of HIF-1α expression by CX-4945 in HuCCT-1 cells, and knockdown of HIF-1α caused a significant reduction of the cell survival. In summary, these results demonstrated that CX-4945 has anti-growth, anti-angiogenic, and pro-apoptotic effects on HuCCT-1 cells, which are mediated through control of CK2, caspase-9/3, DR-4, STAT-3/5, Mcl-1, eIF-2α, and HIF-1α. [ABSTRACT FROM AUTHOR]

In common irrigation systems, sprinklers are mounted with circular nozzles, but innovative noncircular nozzles can save water and energy by improving fragmentation in a low–intermediate pressure irrigation system. In order to investigate the effects of nozzle orifice shapes (circular, square, and equilateral triangular) on droplet characteristics, experiments using high-speed photography and water droplet spectrum measurement were performed. Using ImageJ to observe with the overlapped droplets and using the self-compiled programs of MATLAB to observe the morphology of droplets, we extracted the outlines of droplets. In addition, several empirical formulas for the prediction of droplets were obtained by way of a regression analysis of the experimental data. In particular, the shape coefficient of the nozzle orifice and the operating pressure of the nozzle were added to these formulas as variable factors to make them applicable to a variety of nozzles and working conditions. The results show that with the increase in shape coefficient, the jet atomization intensifies, and the droplets breaking from the jet will be dense and uniform. The velocity distribution of the droplets conforms to exponential functions (R2 > 0.7). The prediction formulas of diameter and kinetic energy were established with coefficients of determination exceeding 0.95. In low pressure conditions, the specific power multiplies at the end of spraying, and the maximum is proportional to the nozzle orifice coefficient. The impact-driven arm compensates for the disadvantage of the noncircular nozzles with the high irrigation-specific power, by producing a wider diameter gradient of droplets. Therefore, innovative sprinklers based on noncircular nozzles can be applied in a low–intermediate pressure system to increase water use efficiency, reduce energy consumption, and reduce costs. [ABSTRACT FROM AUTHOR]

The various environmental pressures could increase the cortisol levels of sturgeon. However, there is a lack of reports on the mechanism by which elevated cortisol levels affect the immune response of Siberian sturgeon. In this study, a high-level cortisol anti-inflammatory state of Siberian sturgeon after co-treatment with LPS and cortisol was constructed and verified in the primary spleen leukocytes. The glucocorticoid receptor (GR) was cloned and its binding site with Ru486 was analyzed. After adding Ru486 to treat the primary spleen leukocytes, the expression of inflammatory cytokines mediated by GR was explored by qRT-PCR. The structure of spleen after LPS + cortisol injection was observed by histopathology and the changes in apoptotic genes were explored by qRT-PCR. Furthermore, the apoptosis of cells after Ru486 treatment was studied by qRT-PCR, DNA ladder, and Hoechst staining. Firstly, co-treatment with LPS and cortisol led to high levels of serum cortisol and glucose within 24 h after treatment. In this situation, the expression of il-10 and tgf-β1 was upregulated in the LPS + cortisol group, while the expression of il-1β and tnf-α was regulated. However, il-1β , tnf-α, il-10 , and tgf-β1 were significantly down-regulated in vitro. Clone analysis of the GR sequence indicated that it is conserved and widely expressed in 18 tissues. Molecular docking simulations suggested that it can bind to cortisol and the GR antagonist Ru486, respectively. Further addition of Ru486 treatment reversed the expression of il-1β , il-10 , tgf-β1 , and JAK-STAT signaling pathway factors. Histopathological observations showed that LPS + cortisol injection causes splenic cell death. Then, the expression of pro-apoptotic genes p53 and caspase3 increases significantly. After Ru486 treatment in spleen leukocytes, the mRNA expression levels of pro-apoptotic-related genes p53 , bax , caspase3 , caspase7 , and caspase9 were significantly downregulated. GR antagonism could attenuate LPS + cortisol-treated splenic leukocyte DNA fragmentation and nuclear morphological changes. Therefore, cortisol can regulate the production of inflammatory cytokines and promote the process of cell apoptosis through the GR of Siberian sturgeon, thereby inhibiting the inflammatory response. • The increased cortisol levels lead to a regulation of anti-inflammatory cytokine and pro-inflammatory cytokine expression. • Ru486 treatment reversed the expression of il-1β , il-10 , and tgf-β1. • GR mediates cortisol - induced apoptosis in splenic leukocytes. [ABSTRACT FROM AUTHOR]