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Objective. The highly conformal carbon-ion radiotherapy is associated with an increased sensitivity of the dose distributions to internal changes in the patient during the treatment course. Hence, monitoring methodologies capable of detecting such changes are of vital importance. We established experimental setup conditions to address the sensitivity of a monitoring approach based on secondary-fragment tracking for detecting clinically motivated air cavity dimensions in a homogeneous head-sized PMMA phantom in 40 mm depth. Approach. The air cavities were positioned within the entrance channel of a treatment field of 50 mm diameter at three lateral positions. The measured secondary-fragment emission profiles were compared to a reference measurement without cavities. The experiments were conducted at the Heidelberg Ion-Beam Therapy Center in Germany at typical doses and dose rates. Main results. Significances above a detectability threshold of 2 σ for the larger cavities (20 mm diameter and 4 mm thickness, and 20 mm diameter and 2 mm thickness) across the entire treatment field. The smallest cavity of 10 mm diameter and 2 mm thickness, which is on the lower limit of clinical interest, could not be detected at any position. We also demonstrated that it is feasible to reconstruct the lateral position of the cavity on average within 2.8 mm, once the cavity is detected. This is sufficient for the clinicians to estimate medical effects of such a cavity and to decide about the need for a control imaging CT. Significance. This investigation defines well-controlled reference conditions for the evaluation of the performance of any kind of treatment monitoring method and its capability to detect internal changes within head-sized objects. Four air cavities with volumes between 0.31 cm3 and 1.26 cm3 were narrowed down around the detectability threshold of this secondary-fragment-based monitoring method. [ABSTRACT FROM AUTHOR]

Symphorema polyandrum belongs to the Lamiaceae family and is locally known as Badichang or mahasindhu. In this study, we performed Soxhlet extraction to prepare methanolic and hydromethanolic extracts, followed by quantification of their total phenolic content and total flavonoid content. Qualitative analysis of both the extracts was conducted to determine the presence of different phytochemicals. In addition, we aimed to identify the important phytochemical constituents in the methanolic extracts of S. polyandrum (SPM) using ultra-performance liquid chromatography hyphenated with high-resolution mass spectrometry (UPLC–ESI–QTOF–MSE). Furthermore, this study investigated the antioxidant, anticancer and anti-inflammatory properties of SPM and its safety profile in the normal fibroblast cell line L929. A colony proliferation assay and a Griess assay were performed to evaluate the effects of SPM on colony formation and nitric oxide (NO) production. A total of 13 important phytochemicals were identified and reported. The methanolic extract of SPM demonstrated significant antioxidant activity. SPM also showed substantial antiproliferative activity on MDA-MB-231 triple-negative breast cancer cells, with an IC50 value of 45.53 ± 1.63 µg/ml, and also reduced the survival of these cancer cells by promoting nuclear fragmentation and condensation without causing harm to normal cells. SPM inhibits the colony formation and reduces the nitric oxide (NO) production. The anti-inflammatory potential of SPM was assessed utilizing the murine alveolar macrophages (J774.A.1) as an in vitro model, and SPM effectively lowered the levels of proinflammatory cytokines such as TNF-α and IL-6. These findings emphasized the antiproliferative potential of SPM to cancer cells, along with its anti-inflammatory, and antioxidant capabilities, indicating the therapeutic efficacy of this medicinal plant. [ABSTRACT FROM AUTHOR]

Many lactic acid bacteria (LAB), known for their human health benefits, are derived from milk and utilized in biotherapeutic applications or for producing valuable nutraceuticals. However, the specific role of milk-associated LAB in biotherapeutics remains underexplored. To address this, eight milk product samples were randomly selected from the Egyptian market, diluted, and then cultured anaerobically on MRS agar. Subsequently, 16 suspected LAB isolates were recovered and underwent rapid preliminary identification. Among these isolates, the Lactobacillus plantarum strain with accession number (OQ547261.1) was identified due to its strong antioxidant activity depending on the DPPH assay, L. plantarum displayed notable antioxidant activities of 71.8% and 93.8% at concentrations of 125–1000 µg/mL, respectively. While ascorbic acid showed lower concentrations of 7.81, 3.9, and 1.95 µg/mL which showed activities of 45.1%, 34.2%, and 27.2%, respectively. The anti-inflammatory efficacy of L. plantarum was evaluated based on its capability to prevent hemolysis induced by hypotonic conditions. At a concentration of 1000 µg/mL, L. plantarum could reduce hemolysis by 97.7%, nearly matching the 99.5% inhibition rate achieved by the standard drug, indomethacin, at an identical concentration. Moreover, L. plantarum exhibited high hemolytic activity at 100 µg/mL (14.3%), which decreased to 1.4% at 1000 µg/mL. The abundance of phenolic acids and flavonoids was determined by high-performance liquid chromatography (HPLC) in L. plantarum. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) demonstrated that L. plantarum increased gene expression of the inflammatory marker TLR2 by 133%, and cellular oxidation markers SOD1 and SOD2 by 65% and 74.2%, respectively, while suppressing CRP expression by 33.3%. These results underscore L. plantarum's exceptional anti-inflammatory and antioxidant activities. Furthermore, L. plantarum induces cancer cell death through necrotic nuclear DNA fragmentation. These findings suggest that L. plantarum is not only suitable for nutraceutical production but also holds potential as a probiotic strain. Future research should focus on enhancing the capacity of this strain across various industries and fostering innovation in multiple fields. [ABSTRACT FROM AUTHOR]

Reduced graphene oxide (rGO) is a potential biomaterial for tissue engineering, photothermal therapy, and drug delivery. However, the hydrophobic nature of rGO results in poor dispersibility in a physiologically relevant aqueous environment, limiting its biomedical applications. To surmount this problem, here, we have developed a silanized derivative of reduced graphene oxide (rSiGO) using 3‐Aminopropyl) triethoxysilane and evaluated its impact on [I] stability of rGO in different aqueous solvents, [ii] drug loading capacities, and [iii] biocompatibility. The physico‐chemical characterization suggested that silanization alters the 2D carbon skeleton of rGO to a certain extent and improves its aqueous dispersibility and drug‐loading capacity. The in vitro cytocompatibility study showed that rSiGO was compatible with the human dermal fibroblasts and murine fibroblast cell lines. It was also found that rSiGO possesses a higher loading capacity for hydrophilic drugs than rGO. The drug‐loaded rSiGO showed higher cytotoxicity against hepatocarcinoma cells by inducing intracellular reactive oxygen species production, apoptosis, and nuclear fragmentation. In vivo systemic toxicity studies in mice showed that the rSiGO is nontoxic at the tested concentration. These results clearly showed that silanization improves the aqueous dispersibility, drug loading capacity, and biocompatibility of rGO making it a better candidate for various biomedical applications. [ABSTRACT FROM AUTHOR]

AIM: To investigate the impact of platycodin D （PD） on the viability, migration, invasion, apoptosis and cell cycle of osteosarcoma cells in vitro, along with its underlying mechanisms. METHODS: Human osteosarcoma cells MG63 and U2OS were divided into control group （0 μmol/L） and PD treatment group （6. 25, 12. 5, 25, 50 and 100 μmol/L, respectively）. Human osteosarcoma cells MG63 and U2OS were categorized into control groups （0 μmol/L PD） and PD treatment groups （6. 25, 12. 5, 25, 50 and 100 μmol/L）. The CCK-8 assay determined cell viability and identified effective treatment concentrations. MG63 （15 μmol/L PD） and U2OS （25 μmol/L PD） were specifically analyzed. Cell scratch and Transwell assays assessed migration and invasion. Hoechst 33342 staining examined nuclear morphological changes. Flow cytometry analyzed cell cycle distribution and apoptosis rate. Western blot measured protein expression levels: cleaved caspase-3, cleaved PARP, c-Jun N-terminal kinase （JNK）, p-JNK, B-cell lymphoma-2 （Bcl2）, Bcl-2-ssociated X protein （BAX）, matrix metalloproteinase 2 （MMP-2）, MMP-9, cyclin-dependent kinase 4 （CDK4）, cyclin D1, CDK1, cyclin B1, extracellular signal-regulated kinase （ERK） and p-ERK. Proteome sequencing of MG63 cells was performed. RESULTS: PD treatment significantly decreased cell survival, scratch healing rate, and invasive cell numbers, while increasing apoptosis rates （P<0. 05）. Morphological changes such as nuclear hyperchromatism and fragmentation were observed in PD-treated cells. PD induced G2/M phase arrest in MG63 and G0/G1 phase arrest in U2OS cells. PD treatment upregulated BAX, cleaved caspase-3, cleaved PARP, and p-JNK/JNK, while downregulating Bcl-2, MMP-2, MMP-9, CDK4, cyclin D1, CDK1, cyclin B1, and p-ERK/ERK（ P<0. 05）. Proteome sequencing revealed PD's involvement in cell division, cell cycle regulation, focal adhesion, apoptosis, and the MAPK signaling pathway. CONCLUSION: PD inhibits cell viability, migration, and invasion of osteosarcoma cells in vitro, while promoting apoptosis and inducing cell cycle arrest. These effects are likely mediated through modulation of the MAPK signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: The objective of this in-vitro study was to explore the pro-apoptotic potential of eugenol (4-allyl-2-methoxyphenol) on Human Leukemia-60 (HL-60) cell line as a potent phytochemical in intensive care setting to leukemia patients. Methodology: After formal approval by all of the respective ethical committees, this study was simultaneously conducted at all respective institutional departments. The study included culturing HL-60 cell line and its treatment with serial concentrations of eugenol for calculation of subsequent IC50 values (half-maximal inhibitory concentration) via MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, expression analysis of gene, comparative analysis of relative gene fold (Intrinsic Biomarkers Caspase-3, Caspase-9) as apoptosis mediator markers followed by RT-qPCR. Cellular apoptotic morphology was confirmed via Hoechst 333258 staining. Results: For HL-60 cell line, the IC50 of eugenol (14.1 uM) showed high gene expression of pro-apoptotic biomarkers (Caspase-3 and Caspase-9). Hoechst 333258 staining showed prominent apoptotic bodies leading to nuclear fragmentations. Conclusion: Eugenol proved to possess robust pro-apoptotic potential leading to diagnostic efficacy against leukemia HL-60 cell line. Further studies would help in identifying key mechanisms by which eugenol exhibits anti-cancer potential against HL-60 cell lines. [ABSTRACT FROM AUTHOR]

The goal of this work is to characterize the secondary neutron spectra produced by 1 GeV/u56Fe beam colliding with a thick cylindric aluminum target and to perform a quantitative comparison with simulated results obtained with Monte Carlo codes. The measurements were performed using extended-range Bonner sphere spectrometers at two positions (15° and 40°) with respect to the beam direction. The secondary radiation field was simulated using four Monte Carlo codes (FLUKA, MCNP6, Geant4 and PHITS) and several physical models of nuclei transport and interaction. Neutron and proton energy distributions were simulated for the experimental measurement positions. The simulated neutron spectra, together with data measured with Bonner sphere spectrometers, after carrying out the correction of the contributions induced by the secondary protons, were used as input for the MAXED spectrum unfolding code to obtain the measured neutron spectra. Unfolded neutron spectra were compared with simulated ones to carry out a quantitative analysis of the performance of the chosen Monte Carlo codes and their corresponding physical models. This comparison showed that, because of experimental uncertainties and physical models, there are no unique solutions for each measurement location, but a range of solutions where the true experimental neutron spectra probably lie. The results showed deviations between 4.23% and 8.42% for some simulated spectra. Regarding the total integral values of neutron fluence and ambient equivalent dose, the unfolded neutron spectra showed deviations lower than 2%. [ABSTRACT FROM AUTHOR]

Colorectal cancer (CRC) is a common and life-threatening neoplastic disease that continues to pose a formidable challenge to global health. The present work was performed to evaluate the anticancer properties of betanin and betanin (BT) loaded starch nanoparticles (S-BT). The BT and S-BT were characterized by DLS, SEM, UV spectroscopy, XPS and FTIR. The cytotoxic effect was assessed by MTT and LDH assay. The apoptotic potential of BT and S-BT was assessed by DCFDA, Rh123, AO/EB and DAPI staining methods. Cell cycle arrest was depicted using flow cytometry. The antimetastatic potential of BT and S-BT was evaluated by wound healing assay. The S-BT showed a spherical morphology with a size of 175 nm. The betanin contained SNPs were found to have strong encapsulation efficiency and favorable release profiles. Both BT and S-BT exhibited cytotoxicity in SW480 cells but S-BT displayed increased cytotoxicity when compared to BT alone. Loss of mitochondrial membrane potential, nuclear fragmentation, chromatin condensation and generation of ROS, all indicative of apoptotic mode of cell death, were revealed by fluorescence imaging. The cells were arrested in the G2M phase. Moreover, both BT and S-BT were able to inhibit the migratory potential of SW480 cells. Overall, our results indicated that both BT and S-BT were able to induce anticancer effects; and, S-BT was found to have increased therapeutic efficacy when compared to BT alone. [ABSTRACT FROM AUTHOR]

This article explores the concept of paraptosis, a type of programmed cell death that is different from apoptosis and necrosis. It discusses the mechanisms of paraptosis and highlights the potential of natural products to induce paraptosis in cancer cells, offering an alternative to traditional cancer therapies. The article also mentions the potential of paraptosis in enhancing tumor immunogenicity and triggering immune responses against cancer cells. Clinical trials are being conducted to test the efficacy and safety of paraptosis in cancer treatment, although challenges such as the lack of rigorous clinical trials and difficulties in large-scale isolation and standardization of natural products exist. Overall, the article provides valuable insights into the field of paraptosis and its potential applications in cancer therapy. [Extracted from the article]

Neutron capture enhanced particle therapy (NCEPT) is a proposed augmentation of charged particle therapy that exploits thermal neutrons generated internally, within the treatment volume via nuclear fragmentation, to deliver a biochemically targeted radiation dose to cancer cells. This work is the first experimental demonstration of NCEPT, performed using both carbon and helium ion beams with 2 different targeted neutron capture agents (NCAs). Human glioblastoma cells (T98G) were irradiated by carbon and helium ion beams in the presence of NCAs [10B]-BPA and [157Gd]-DOTA-TPP. Cells were positioned within a polymethyl methacrylate phantom either laterally adjacent to or within a 100 × 100 × 60 mm spread out Bragg peak (SOBP). The effect of NCAs and location relative to the SOBP on the cells was measured by cell growth and survival assays in 6 independent experiments. Neutron fluence within the phantom was characterized by quantifying the neutron activation of gold foil. Cells placed inside the treatment volume reached 10% survival by 2 Gy of carbon or 2 to 3 Gy of helium in the presence of NCAs compared with 5 Gy of carbon and 7 Gy of helium with no NCA. Cells placed adjacent to the treatment volume showed a dose-dependent decrease in cell growth when treated with NCAs, reaching 10% survival by 6 Gy of carbon or helium (to the treatment volume), compared with no detectable effect on cells without NCA. The mean thermal neutron fluence at the center of the SOBP was approximately 2.2 × 109 n/cm2/Gy (relative biological effectiveness) for the carbon beam and 5.8 × 109 n/cm2/Gy (relative biological effectiveness) for the helium beam and gradually decreased in all directions. The addition of NCAs to cancer cells during carbon and helium beam irradiation has a measurable effect on cell survival and growth in vitro. Through the capture of internally generated neutrons, NCEPT introduces the concept of a biochemically targeted radiation dose to charged particle therapy. NCEPT enables the established pharmaceuticals and concepts of neutron capture therapy to be applied to a wider range of deeply situated and diffuse tumors, by targeting this dose to microinfiltrates and cells outside of defined treatment regions. These results also demonstrate the potential for NCEPT to provide an increased dose to tumor tissue within the treatment volume, with a reduction in radiation doses to off-target tissue. [ABSTRACT FROM AUTHOR]

Since the adoption of the nuclear Non-Proliferation Treaty (NPT), the order founded on the treaty has evolved into an increasingly fragmented regime complex. Fragmentation has resulted from forum-shifting initiated both at the top and at the bottom of the nuclear hierarchy: both the United States, with varying partners, and coalitions of small-and medium-sized non-nuclear states have repeatedly moved rule-making on nuclear issues to fora outside the NPT, adding partly conflicting institutions to the complex. To understand this dynamic, I propose a sociological perspective that highlights states' positional struggles in a multidimensionally stratified international society. Drawing on Bourdieusian global fields theory, I argue that both dominant and weak states use forum-shifting to manipulate exchange rates between different (material, institutional and social) forms of capital they possess. Thus, they seek to protect their positions in global hierarchies within and beyond the nuclear field when they perceive these hierarchies as being challenged by material or institutional power shifts. [ABSTRACT FROM AUTHOR]

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Nonclinical studies of test articles (TAs) in nonhuman primates are often designed to assess both biodistribution and toxicity. For this purpose, studies commonly use intravenous perfusion of ice-cold (2°C-8°C) saline to facilitate measurements of TA-associated nucleic acids and proteins, after which tissues undergo later fixation by immersion for histological processing and microscopic evaluation. Intriguingly, minimal apoptosis/single cell necrosis (A/SCN) of randomly distributed neural cells is evident in the cerebral cortex and less often the hippocampus in animals from all groups, including vehicle-treated controls. Affected cells exhibit end-stage features such as cytoplasmic hypereosinophilia, nuclear condensation or fragmentation, and shape distortions, so their lineage(s) generally cannot be defined; classical apoptotic bodies are exceedingly rare. In addition, A/SCN is not accompanied by glial reactions, leukocyte infiltration/inflammation, or other parenchymal changes. The severity is minimal in controls but may be slightly exacerbated (to mild) by TA that accumulate in neural cells. One plausible hypothesis explaining this A/SCN exacerbation is that cold shock (perhaps complicated by concurrent tissue acidity and hypoxia) drives still-viable but TA-stressed cells to launch a self-directed death program. Taken together, these observations indicate that A/SCN in brain processed by cold saline perfusion with delayed immersion fixation represents a procedural artifact and not a TA-related lesion. [ABSTRACT FROM AUTHOR]

Hepatocyte organoids (HOs) have superior hepatic functions to cholangiocyte-derived organoids but suffer from shorter lifespans. To counteract this, we co-cultured pig HOs with adipose-derived mesenchymal stem cells (A-MSCs) and performed transcriptome analysis. The results revealed that A-MSCs enhanced the collagen synthesis pathways, which are crucial for maintaining the three-dimensional structure and extracellular matrix synthesis of the organoids. A-MSCs also increased the expression of liver progenitor cell markers (KRT7, SPP1, LGR5+, and TERT). To explore HOs as a liver disease model, we exposed them to alcohol to create an alcoholic liver injury (ALI) model. The co-culture of HOs with A-MSCs inhibited the apoptosis of hepatocytes and reduced lipid accumulation of HOs. Furthermore, varying ethanol concentrations (0–400 mM) and single-versus-daily exposure to HOs showed that daily exposure significantly increased the level of PLIN2, a lipid storage marker, while decreasing CYP2E1 and increasing CYP1A2 levels, suggesting that CYP1A2 may play a critical role in alcohol detoxification during short-term exposure. Moreover, daily alcohol exposure led to excessive lipid accumulation and nuclear fragmentation in HOs cultured alone. These findings indicate that HOs mimic in vivo liver regeneration, establishing them as a valuable model for studying liver diseases, such as ALI. [ABSTRACT FROM AUTHOR]

Background: Triple‐negative breast cancer (TNBC) is the most aggressive and chemo‐resistant form of breast cancer subtype, and chemotherapy is a vital treatment option for that. Paclitaxel is an effective chemo drug for TNBC. However, in clinical settings, paclitaxel has adverse side effects. The synergistic combination is the most promising method for overcoming undesirable toxicity and achieving a beneficial therapeutic outcome. Previous reports, including our study, showed certain anticancer potential of epoxyazadiradione (EAD), the neem limonoid, in different types of cancer cells, including TNBC. Objective: This study was designed to investigate the possible synergistic effects of EAD and paclitaxel against TNBC cells. Methods: We examined the effects of EAD and paclitaxel alone and in combination in MDA‐MB 231 cells, and the percentage cytotoxicity was used to calculate synergism. Characteristic apoptotic changes were observed by visualizing cellular morphology, nuclear fragmentation and membrane integrity. We further estimated anti‐migratory potential of experimental compounds by wound healing assay. The reduction in inflammation during combinatorial treatment was evaluated by observing NF‐κB translocation. Results: The combined treatment with EAD (5 μM) and paclitaxel (5 nM), which were used at doses lower than their individual IC50 concentrations, showed a synergistic effect in MDA‐MB‐231 cells. This combination effectively induced apoptosis and antimigration and reduced the inflammatory reactions induced by the higher dose of paclitaxel. Conclusion: To conclude, EAD could be the drug of choice for combined treatment with paclitaxel in a chemotherapy regimen. [ABSTRACT FROM AUTHOR]

This study delves into the pathogenesis of virulent genotype VII strains of the Newcastle disease virus (NDV), focusing on experimentally infected birds. Predominant and consistent lesions observed include bursal atrophy and extensive depletion of all lymphoid tissues. Immunohistochemistry (IHC) analysis, targeting apoptosis (Caspase-3), necroptosis (MLKL), and NDV markers, indicates that bursal atrophy is linked to a non-apoptotic programmed cell death pathway known as "necroptosis". Repair assisted damage detection (RADD) of the bursa reveal oxidative DNA damage patterns consistent with programmed cell death, aligning with MLKL expression. Contrastingly, in the spleen, our findings suggest that necrosis (non-programmed cell death) predominantly contributes to lymphoid depletion. This conclusion is supported by evidence of karyorrhexis, fibrinous inflammation, RADD analyses, and IHC. Moreover, in addition to being pathogenic in its own right, NDV caused extensive and rapid lymphoid depletion that should be expected to contribute to profound immunosuppression. The elucidation of necroptosis in NDV-infected chickens provides a good rationale to investigate this mechanism in other paramyxoviral diseases such as human measles. [ABSTRACT FROM AUTHOR]

The miCOR, FDA-approved in 2022, revolutionizes cataract surgery as a handheld, disposable, subsonic lens extraction device providing a safe, cost-effective phacoemulsification-free solution with a smaller operating room footprint. Purpose: To investigate the intraoperative performance and lens fragmentation efficacy of a non-cavitating handheld lensectomy system in mild, moderate, and severe cataract. Setting: Ambulatory surgical centers. Design: Retrospective consecutive case series. Methods: 665 consecutive eyes underwent cataract surgery by 12 surgeons using a new handheld non-cavitating lensectomy system for nuclear fragmentations and extraction. Intraoperative measurements included surgical time, miLOOP pretreatment, and irrigation fluid use. Results: Of the 665 eyes, 38 (6%), 468 (70%), 126 (19%), and 33 (5%) were of grade 1, 2, 3, and 4 nuclear densities, respectively, as graded by the surgeon intraoperatively. Successful nuclear fragmentation, lens extraction, and cortical removal were achieved in all eyes. Total nucleus fragmentation and extraction times were 70.1 seconds, 100.3 seconds, 132.6 seconds, and 287.9 seconds for grades 1, 2, 3, and 4, respectively (P <.001). In addition, irrigation and aspiration cortical removal times were 64.1 seconds, 51.1 seconds, 48.5 seconds, and 59.0 seconds, respectively (P =.14). There was a low rate of capsular tear (3 cases in 665 surgeries, 0.45%) and no other emergent adverse events. Conclusions: The miCOR handheld non-cavitating lensectomy system demonstrated nuclear fragmentation and extraction in the absence of intraocular cavitation across all grades of nuclear densities. [ABSTRACT FROM AUTHOR]

Background: While the kinetics of human sperm nuclear DNA fragmentation (SDF‐nDNA) following ejaculation have been described, the dynamics and relationships of mitochondrial DNA copy number per spermatozoon (mtDNAcn) and fragmentation (SDF‐mtDNA) remain unexplored. Objectives: To compare post‐ejaculatory kinetics of mtDNAcn, SDF‐mtDNA and SDF‐nDNA, global, single‐strand DNA breaks (SDF‐SSBs) and double‐strand DNA breaks (SDF‐DSBs) in normozoospermic and non‐normozoospermic samples. Materials and methods: 28 normozoospermic and 43 non‐normozoospermic ejaculates were evaluated at 0, 6, 24 and 48 h of incubation in vitro. SDF‐nDNA was determined by sperm chromatin dispersion (SCD) assays. mtDNAcn and SDF‐mtDNA were analysed by dPCR. Results: SDF‐nDNA‐global values increased as a consequence of quadratic SDF‐SSBs and linear SDF‐DSBs kinetics. Non‐normozoospermic samples showed a slower SDF‐global rate between 6–24 h, due to lesser SSBs production. Regarding SDF‐DSBs, non‐normozoospermic samples exhibited a faster initial increase rate, followed by a slower final increment. The mtDNAcn median value decreased linearly, being 3.2× higher in non‐normozoospermics at all time points; mtDNAcn in both cohorts reduced to half of the baseline by 48 h. mtDNAcn was identified as a risk factor for discriminating non‐normozoospermia, a finding that was further strengthen when combined with SDF‐Global or SDF‐DSBs values. SDF‐mtDNA frequencies were identical, increasing over time correspondingly in both cohorts. The mtDNA fragmentation rate was initially fast, decreasing progressively with time for both cohorts; half of the initially unfragmented copies were fragmented after 48 h. Rates of mtDNAcn loss and SDF‐mtDNA increase were only marginally correlated with the rates of nuclear fragmentation. Conclusion: mtDNA fragmentation and loss occur post ejaculation. Their dynamics are likely to be associated with different and/or uncoupled mechanisms to that which cause nuclear DNA fragmentation. Our results indicate that while mtDNA fragmentation is not influenced by the sperm quality, the number of copies of sperm mtDNAcn can potentially serve as a risk factor for predicting non‐normozoospermia. [ABSTRACT FROM AUTHOR]

o‐Aminoazotoluene (OAT), capable of photoisomerization, and o‐vanillin, a potent comutagen, have been used to synthesize a new ligand, HL, [C6H4(CH3)N=NC6H3(CH3)N=CHC6H3(OH)(OCH3)], which, upon complexation with copper(II), results in a new copper(II) complex‐Cu(L)2, [Cu{C6H4(CH3)N=NC6H3(CH3)N=CHC6H3(OCH3)O}2]. Both HL and Cu(L)2 have been characterized by single‐crystal X‐ray diffraction measurements along with other analytical techniques, for example, IR spectroscopy, NMR spectroscopy, UV–Vis spectroscopy, elemental analysis and mass spectrometry. These compounds were tested with different in vitro anticancer assay as well as with in silico studies. A comparative study has been demonstrated on anticancer activity of HL and Cu(L)2 with the ligand HAZ, {C6H5N=NC6H4N=CHC6H3(OH)(OCH3)} and its Cu‐complex, [Cu(AZ)2, Cu{C6H5N=NC6H4N=CHC6H3(OCH3)O}2]. The sensing properties of HAZ, along with the synthesis and structural properties of both HAZ and Cu(AZ)2, have been reported by our group earlier. Cytotoxicity measurements on MCF7 cell lines show that Cu(L)2 and Cu(AZ)2 have higher anticancer activity than their corresponding ligands. The apoptotic effect of Cu‐complex was studied through nuclear fragmentation assay and AO/EB dual‐staining assay on MCF7 cell line. The IC50 value of Cu(L)2 in 0.01% DMSO/water after 24‐h treatment was found 4.2 μM, which is one of the lowest values with this response time compared to the other analogous anticancer compounds. Finally, we have evaluated the expression of hERα protein with respect to Cu‐complexes, and it was observed that Cu(L)2 caused more down‐regulation of hERα as compared to Cu(AZ)2. [ABSTRACT FROM AUTHOR]

Usnic acid (UA) is a unique bioactive substance in lichen with potential anticancer properties. Recently, we have reported that UA can reduce 7,12‐dimethylbenz[a] anthracene‐induced oral carcinogenesis by inhibiting oxidative stress, inflammation, and cell proliferation in a male golden Syrian hamster in vivo model. The present study aims to explore the relevant mechanism of cell death induced by UA on human oral carcinoma (KB) cell line in an in vitro model. We found that UA can induce apoptosis (cell death) in KB cells by decreasing cell viability, increasing the production of reactive oxygen species (ROS), depolarizing mitochondrial membrane potential (MMP) levels, causing nuclear fragmentation, altering apoptotic morphology, and causing excessive DNA damage. Additionally, UA inhibits the expression of Bcl‐2, a protein that promotes cell survival, while increasing the expression of p53, Bax, Cytochrome‐c, Caspase‐9, and 3 proteins in KB cells. UA also inhibits the expression of nuclear factor‐κB (NF‐κB), a protein that mediates the activation of pro‐inflammatory cytokines such as TNF‐α and IL‐6, in KB cells. Furthermore, UA promotes apoptosis by enhancing the mitochondrial‐mediated apoptotic mechanism through oxidative stress, depletion of cellular antioxidants, and an inflammatory response. Ultimately, the findings of this study suggest that UA may have potential as an anticancer therapeutic agent for oral cancer treatments. Significance statement: Usnic acid (UA) has been found to inhibit the growth of cancerous cells and induce the generation of ROS (reactive oxygen species) by activating apoptotic signaling pathways in KB cells. More importantly, the present study suggests that UA Prevent cytotoxicity depletion of antioxidants, induce apoptotic molecules, and downregulation of the inflammatory response in KB cells. Due to its antioxidant effects, UA could potentially be used to develop new anticarcinoma agents, making it a promising novel therapeutic candidate for treating human oral cancer. [ABSTRACT FROM AUTHOR]

FLACS with customized nuclear fragmentation patterns resulted in significantly less intraoperative phacoemulsification time, surgical time, and endothelial cell loss compared with conventional phacoemulsification for dense cataracts. Purpose: To compare the safety, effectiveness, and changes in endothelial cell density (ECD) after standard phacoemulsification and femtosecond laser-assisted cataract surgery (FLACS) in dense cataracts (Lens Opacities Classification System III grade 3.0 NO and above). Setting: Nethradhama Superspeciality Eye Hospital, Bangalore, India. Design: Prospective, randomized, comparative study. Method: 100 eyes of 100 patients were randomly assigned to either conventional phacoemulsification surgery (CPS) or FLACS (LENSAR Laser System) using predefined patterns of nuclear fragmentations. Measured outcomes included intraoperative phacoemulsification time, irrigation fluid volume, surgical time, and complication rates. ECD was evaluated at baseline, 2 weeks, and 6 months postoperatively. Results: The CPS group had significantly higher mean total phacoemulsification time (11.17 ± 6.5 seconds vs 8.03 ± 3.77 seconds), effective phacoemulsification time (6.14 ± 3.62 seconds vs 4.42 ± 2.07 seconds), and total surgical time (8.18 ± 2.36 minutes vs 7.11 ± 1.55 minutes) than the FLACS group (P <.001). Volume of irrigation fluid was comparable between the 2 groups (CPS, 36.7 ± 12.18 mL; FLACS, 38.64 ± 13.73 mL (P =.45). Mean corneal edema score and central corneal thickness values on postoperative day 1 were significantly higher for the CPS group. At 6 months, % ECD loss was significantly higher in the CPS group (16.08%) vs the FLACS group (12.8%) (P <.001). 3 eyes in the CPS group had wound burns of varying severity. Conclusions: FLACS with customized nuclear fragmentation patterns resulted in significantly less intraoperative phacoemulsification time, surgical time, and endothelial cell loss compared with conventional phacoemulsification when treating dense cataracts. Trial Registration Number: CTRI/2021/12/038887. [ABSTRACT FROM AUTHOR]

Objective: Mitosis karyorrhexis index (MKI) can reflect the proliferation status of neuroblastoma cells. This study aimed to investigate the contrast-enhanced computed tomography (CECT) radiomics features associated with the MKI status in neuroblastoma. Materials and methods: 246 neuroblastoma patients were retrospectively included and divided into three groups: low-MKI, intermediate-MKI, and high-MKI. They were randomly stratified into a training set and a testing set at a ratio of 8:2. Tumor regions of interest were delineated on arterial-phase CECT images, and radiomics features were extracted. After reducing the dimensionality of the radiomics features, a random forest algorithm was employed to establish a three-class classification model to predict MKI status. Results: The classification model consisted of 5 radiomics features. The mean area under the curve (AUC) of the classification model was 0.916 (95% confidence interval (CI) 0.913–0.921) in the training set and 0.858 (95% CI 0.841–0.864) in the testing set. Specifically, the classification model achieved AUCs of 0.928 (95% CI 0.927–0.934), 0.915 (95% CI 0.912–0.919), and 0.901 (95% CI 0.900–0.909) for predicting low-MKI, intermediate-MKI, and high-MKI, respectively, in the training set. In the testing set, the classification model achieved AUCs of 0.873 (95% CI 0.859–0.882), 0.860 (95% CI 0.852–0.872), and 0.820 (95% CI 0.813–0.839) for predicting low-MKI, intermediate-MKI, and high-MKI, respectively. Conclusions: CECT radiomics features were found to be correlated with MKI status and are helpful for reflecting the proliferation status of neuroblastoma cells. [ABSTRACT FROM AUTHOR]

The goal of this work is to characterize the secondary neutron spectra produced by 1 GeV/u56Fe beam colliding with a thick cylindric aluminum target and to perform a quantitative comparison with simulated results obtained with Monte Carlo codes. The measurements were performed using extended-range Bonner sphere spectrometers at two positions (15° and 40°) with respect to the beam direction. The secondary radiation field was simulated using four Monte Carlo codes (FLUKA, MCNP6, Geant4 and PHITS) and several physical models of nuclei transport and interaction. Neutron and proton energy distributions were simulated for the experimental measurement positions. The simulated neutron spectra, together with data measured with Bonner sphere spectrometers, after carrying out the correction of the contributions induced by the secondary protons, were used as input for the MAXED spectrum unfolding code to obtain the measured neutron spectra. Unfolded neutron spectra were compared with simulated ones to carry out a quantitative analysis of the performance of the chosen Monte Carlo codes and their corresponding physical models. This comparison showed that, because of experimental uncertainties and physical models, there are no unique solutions for each measurement location, but a range of solutions where the true experimental neutron spectra probably lie. The results showed deviations between 4.23% and 8.42% for some simulated spectra. Regarding the total integral values of neutron fluence and ambient equivalent dose, the unfolded neutron spectra showed deviations lower than 2%.

Background: While it is common to clinically evaluate sperm nuclear DNA fragmentation, less attention has been given to sperm mitochondrial DNA. Recently, a digital PCR assay has allowed accurate estimation of the proportion of fragmented mtDNA molecules and relative copy number. Objectives: To determine the correlation of classical sperm parameters, average mtDNA copies per spermatozoon and the level of mtDNA fragmentation (SDF‐mtDNA) to that of nuclear DNA fragmentation (SDF‐nDNA), measured as the proportion of global, single‐strand DNA (SDF‐SSBs) and double‐strand DNA breaks (SDF‐DSBs). To determine whether the level of nuclear and mitochondrial DNA fragmentation and/or copy number can differentiate normozoospermic from non‐normozoospermic samples. Materials and methods: Ejaculates from 29 normozoospermic and 43 non‐normozoospermic were evaluated. SDF was determined using the sperm chromatin dispersion assay. mtDNA copy number and SDF‐mtDNA were analyzed using digital PCR assays. Results: Relative mtDNA copy increased as sperm concentration or motility decreased, or abnormal morphology increased. Unlike SDF‐mtDNA, mtDNA copy number was not correlated with SDF‐nDNA. SDF‐mtDNA increased as the concentration or proportion of non‐vital sperm increased; the higher the mtDNA copy number, the lower the level of fragmentation. Non‐normozoospermic samples showed double the level of SDF‐nDNA compared to normozoospermic (median 25.00 vs. 13.67). mtDNA copy number per spermatozoon was 3× higher in non‐normozoospermic ejaculates (median 16.06 vs. 4.99). Although logistic regression revealed SDF‐Global and mtDNA copy number as independent risk factors for non‐normozoospermia, when SDF‐Global and mtDNA copy number were combined, ROC curve analysis resulted in an even stronger discriminatory ability for predicting the probability of non‐normozoospermia (AUC = 0.85, 95% CI 0.76–0.94, p < 0.001). Conclusion: High‐quality ejaculates show lower nuclear SDF and retain less mtDNA copies, with approximately half of them fragmented, so that the absolute number of non‐fragmented mtDNA molecules per spermatozoon is extremely low. [ABSTRACT FROM AUTHOR]

A library of 3-(5-[(substituted benzyl)sulfanyl]-1,3,4-oxadiazol-2-yl)-N-substituted pyridine-2-amines and 2-[(5-{2-[(substituted phenyl)amino]pyridin-3-yl}-2,3-dihydro-1,3,4-oxadiazol-2-yl)sulfanyl]-N-substituted phenylacetamides were synthesized by the multistage procedure, starting from 2-chloronicotinic acid. The newly synthesized compounds were tested for in vitro cytotoxicity against the MCF-7 breast cancer cell line. Significant cell death rates were demonstrated by all the test compounds in a concentration-dependent manner. The active compounds N-(4-fluorophenyl)-3-{5-[(4-nitrobenzyl)sulfanyl]-1,3,4-oxadiazol-2-yl}pyridin-2-amine and 2-[(5-{2-[(3-chloro-4-fluorophenyl)amino]pyridin-3-yl}-1,3,4-oxadiazol-2-yl)sulfanyl]-N-(2,4-dichlorophenyl)-acetamide were further subjected to DAPI staining to assess their effect on nuclear fragmentation. Additionally, the synthesized compounds were screened for anti-inflammatory and antioxidant activities, yielding promising results. A molecular docking study was conducted to assess the binding affinities of the synthesized compounds to PDB 3ERT (human estrogen receptor-α in complex with 4-hydroxytamoxifen). For all the compounds, good binding energies with the target protein were predicted. An ADME screening showed that the majority of the synthesized compounds have good pharmacokinetic profiles. [ABSTRACT FROM AUTHOR]

This paper describes events of anomalously high energy transfer to a micro-object by fragments of nuclei generated in nuclear interactions in the environment on board a spacecraft in flight in low-Earth orbit. An algorithm has been developed that allows for the calculation of the absorbed energy from one or more fragments - products of nuclear interaction. With this algorithm the energy distributions for a spherical micro-volume in an aqueous medium were calculated. And the resulting absorbed energy spectra from nuclear fragments and from primary cosmic rays were compared. The role of nuclear interactions in events of large energy transfers in micro-objects in the field of primary cosmic radiation has been evaluated. The calculations performed in this study showed that the energy in a micro-volume from nuclear events can be several times higher compared to the energy imparted by primary space radiation. [ABSTRACT FROM AUTHOR]

Cell viability and cytotoxicity assays play a crucial role in drug screening and evaluating the cytotoxic effects of various chemicals. The quantification of cell viability and proliferation serves as the cornerstone for numerous in vitro assays that assess cellular responses to external factors. In the last decade, several studies have developed guidelines for defining and interpreting cell viability and cytotoxicity based on morphological, biochemical, and functional perspectives. As this domain continues to experience ongoing growth, revealing new mechanisms orchestrating diverse cell cytotoxicity pathways, we suggest a revised classification for multiple assays employed in evaluating cell viability and cell death. This classification is rooted in the cellular compartment and/or biochemical element involved, with a specific focus on mechanistic and essential aspects of the process. The assays are founded on diverse cell functions, encompassing metabolic activity, enzyme activity, cell membrane permeability and integrity, adenosine 5′‐triphosphate content, cell adherence, reduction equivalents, dye inclusion or exclusion, constitutive protease activity, colony formation, DNA fragmentation and nuclear splitting. These assays present straightforward, reliable, sensitive, reproducible, cost‐effective, and high‐throughput approaches for appraising the effects of newly formulated chemotherapeutic biomolecules on the cell survival during the drug development process. Significance statement: Various perspectives exist regarding the definitions of cell viability, cell death, and cytotoxicity. This review article aims to provide an up‐to‐date classification of commonly used in vitro cytotoxicity assays. The goal is to provide an in‐depth exploration of their underlying principles, the biochemical components involved, the specific cellular compartments targeted, and a comprehensive analysis of their respective strengths and limitations. Potential false positive results exists in cytotoxicity assessments. For instance, when there is a decrease in viability following a proliferation assay, the utilization of mandatory cell death confirmation assays. Therefore, meticulous consideration of the specific mechanisms underlying cell death, along with the incorporation of appropriate confirmation assays, becomes imperative in ensuring the accuracy and reliability of cytotoxicity assessments. [ABSTRACT FROM AUTHOR]

The present experiments are aimed to examine the effect of copper nanoparticles supported on charcoal (CuNPs/C), growth factor betacellulin (BTC) and their interrelationships in the control of ovarian cell functions. Porcine ovarian granulosa cells were cultured in the presence of CuNPs/C (0, 1, 10 or 100 ng/ml), BTC (100 ng/ml) and the combination of both, CuNPs/C + BTC. Markers of cell proliferation (BrDU incorporation), of the S -phase (PCNA) and G-phase (cyclin B1) of the cell cycle, markers of extrinsic (nuclear DNA fragmentation) and cytoplasmic/mitochondrial apoptosis (bax and caspase 3), and the release of progesterone and estradiol were assessed by BrDU test, TUNEL, quantitative immunocytochemistry and ELISA. Both CuNPs/C and BTC, when added alone, increased the expression of all the markers of cell proliferation, reduced the expression of all apoptosis markers and stimulated progesterone and estradiol release. Moreover, BTC was able to promote the CuNPs/C action on the accumulation of PCNA, cyclin B1, bax and estradiol output. These observations demonstrate the stimulatory action of both CuNPs/C and BTC on ovarian cell functions, as well as the ability of BTC to promote the action of CuNPs/C on ovarian cell functions. [Display omitted] • Copper nanoparticles on charcoal and betacellulin, independently or in combination, can be biostimulators of porcine ovarian cell functions. [ABSTRACT FROM AUTHOR]

Background and Aim: Colorectal cancer stands as a frequently occurring fatal disease for several decades and the use of nanoparticles has long been explored in cancer treatments. Engineering the size and shape to attain an optimal efficacy of the nanoparticles and using a specific plant resource to enhance its compatibility to other healthy cell is a propitious approach. The present study investigates the anti-cancer activity of CuO NP synthesized from Aegle marmelos leaf extract against human colorectal cancer. Materials and Methods: The CuO NPs (Nanoparticles) exhibited promising characteristics with UV and SEM-EDAX, its photochemical constituents were analyzed with FTIR, and the geometry of the synthesized nano-particles was studied using Zeta potential and particle size analyzer. The cell viability was scrutinized via trypan blue assay and further toxicity was determined by cell morphology and several in vitro analyses where the MTT assay presented that the 25 µg/mL of CuO NPs as the IC50 concentration. RESULTS: The nuclear fragmentation was studied by DAPI staining which resulted in increased fluorescence of the treated cells indicating the robust effect of the CuO NPs and the mitochondrial damage was monitored through MMP analysis with declining fluorescence. The most vital aspect of understanding cancer pathogenesis is oxidative stress which was evaluated by ROS, NO, and LPO with favorable outcome. Conclusion: The cell viability and ROS assay coalesce to suggest that CuO NPs indeed induce apoptosis and it is evident that Biogenically synthesized CuO NPs contain anti-proliferative potential and promotes apoptosis. [ABSTRACT FROM AUTHOR]

Normoxic inactivation of prolyl hydroxylase-2 (PHD-2) in tumour microenvironment paves the way for cancer cells to thrive under the influence of HIF-1a and NF-κB. Henceforth, the present study is aimed to identify small molecule activators of PHD-2. A virtual screening was conducted on a library consisting of 265,242 chemical compounds, with the objective of identifying molecules that exhibit structural similarities to the furan chalcone scaffold. Further, PHD-2 activation potential of screened compound was determined using in vitro 2-oxoglutarate assay. The cytotoxic activity and apoptotic potential of screened compound was determined using various staining techniques, including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, 4',6-diamidino-2-phenylindole (DAPI), 1,1',3,3'-tetraethylbenzimi-dazolylcarbocyanine iodide (JC-1), and acridine orange/ethidium bromide (AO/EB), against MCF-7 cells. 7,12-Dimethylbenz[a]anthracene (DMBA) model of mammary gland cancer was used to study the in vivo antineoplastic efficacy of screened compound. [(E)-1-(4-fluorophenyl)- 3-(furan-2-yl) prop-2-en-1-one] (BBAP-7) was screened and validated as a PHD-2 activator by an in vitro 2-oxo-glutarate assay. The IC50 of BBAP-7 on MCF-7 cells is 18.84 μM. AO/EB and DAPI staining showed nuclear fragmentation, blebbing and condensation in MCF-7 cells following BBAP-7 treatment. The red-to-green intensity ratio of JC-1 stained MCF-7 cells decreased after BBAP-7 treatment, indicating mitochondrial-mediated apoptosis. DMBA caused mammary gland dysplasia, duct hyperplasia and ductal carcinoma in situ. Carmine staining, histopathology, and scanning electron microscopy demonstrated that BBAP-7, alone or with tirapazamine, restored mammary gland surface morphology and structural integrity. Additionally, BBAP-7 therapy significantly reduced oxidative stress and glycolysis. The findings reveal that BBAP-7 activates PHD-2, making it a promising anticancer drug. [ABSTRACT FROM AUTHOR]

Introduction: Chlorogenic acid, the primary active component in Chinese medicines like honeysuckle, exhibits anti-inflammatory and antiviral effects. It has been demonstrated that chlorogenic acid effectively prevents and treats Duck enteritis virus (DEV) infection. This study aims to further elucidate the mechanism by which chlorogenic acid prevents DEV infection. Methods: Duck embryo fibroblast (DEF) cells were pre-treated with chlorogenic acid before being infected with DEV. Cell samples were collected at different time points for transcriptomic sequencing, while qPCR was used to detect the proliferation of DEV. Additionally, 30-day-old ducks were treated with chlorogenic acid, and their lymphoid organs were harvested for histopathological sections to observe pathological damage. The proliferation of DEV in the lymphoid organs was also detected using qPCR Based on the transcriptomic sequencing results, NF-κB1 gene was silenced by RNAi technology to analyze the effect of NF-κB1 gene on DEV proliferation. Results: Compared to the viral infection group, DEF cells in the chlorogenic acid intervention group exhibited significantly reduced DEV load (P < 0.05). Transcriptomic sequencing results suggested that chlorogenic acid inhibited DEV proliferation in DEF cells by regulating NF-κB signaling pathway. The results of RNAi silencing suggested that in the three treatment groups, compared with the DEV experimental group, there was no significant difference in the effect of pre-transfection after transfection on DEV proliferation, while both the pre-transfection after transfection and the simultaneous transfection group showed significant inhibition on DEV proliferation Furthermore, compared to the virus infection group, ducks in the chlorogenic acid intervention group showed significantly decreased DEV load in their lymphoid organs (P < 0.05), along with alleviated pathological damage such as nuclear pyretosis and nuclear fragmentation. Conclusions: Chlorogenic acid effectively inhibits DEV proliferation in DEF and duck lymphatic organs, mitigates viral-induced pathological damage, and provides a theoretical basis for screening targeted drugs against DEV. [ABSTRACT FROM AUTHOR]

The health effects of galactic cosmic radiation are a serious impediment to crewed exploration of the solar system. OLTARIS, an interface for the 3DHZETRN deterministic radiation transport code, was used to assess the response of aerospace materials to this constant radiation exposure. Traditional aerospace structural materials like aluminum can, after a certain mass, increase the health effects of such radiation. However, materials with lower atomic mass may mitigate this build-up in secondary radiation with increasing areal density. As such, lower atomic mass structural alloys of magnesium and magnesium–lithium are promising candidates. These alloys may reduce the mass of structures when substituted for aluminum alloys. Reinforcement with boron carbide could further reduce atomic mass while also improving the mechanical properties of such lightweight alloys. This study found that the lower atomic mass of these materials increased nuclear fragmentation upon cosmic radiation interactions, leading to a softening of the secondary (neutron) radiation spectra. This softened spectra reduced the effective dose equivalent, a measure of health effects, for magnesium(-lithium) alloys and their boron carbide-reinforced composites when compared to aluminum. • Mg and Mg-14wt%Li mitigate the build-up in secondary space radiation at high areal densities. • B 4 C addition to metals also mitigates the build-up in secondary space radiation. • Lower atomic masses increase fragmentation and soften the secondary neutron energy spectra. [ABSTRACT FROM AUTHOR]

This study aimed to compare the morphometry of horse and mule embryos. The study's hypothesis was that the micronuclei and nuclear fragmentation indexes are higher in mule embryos than in horse embryos. Twenty-two mares were randomly assigned in a crossover design to receive semen from a horse and a donkey; thirteen horse and thirteen mule embryos were obtained. Embryos were recovered eight days post-ovulation and classified according to the stage of development and quality with a score from 1 (excellent) to 4 (degenerate). Embryos were stained with Hoechst33342, and images were acquired with a fluorescence microscope. Nuclei were categorized as compact, mitotic, or fragmented; the fragmented and mitotic indexes were calculated based on their proportion over the total amount of nuclei counted. Embryo size and nuclear morphometry were assessed through ImageJ. Data analyses were carried out with GraphPad using ANOVA and T-test; significance was set at P < 0.05. The number of positive flushes in cycles bred with donkey or stallion semen did not differ when compared per cycle or per ovulation (13 vs. 12) (P > 0.05). One set of twins was recovered from a mare bred to the stallion that had a double ovulation; a mule and horse embryos were both recovered from eight mares. There was no difference in size between mule and horse embryos (915.5 ± 288 μm vs. 575.8 ± 69.6 μm) (P > 0.05) size of the study. The mule embryos scored between grade 1 (n = 9) and grade 2 (n = 4); similarly, the horse embryos scored between grade 1 (n = 6) and grade 2 (n = 7). The evaluation of the nuclear morphometry revealed that horse and mule embryos have a similar number of compact nuclei per sector (148.7 ± 6.8 nuclei/sector in mule embryos vs. 156.5 ± 8.5 nuclei/sector in horse embryos) (P > 0.05); however, the number of mitotic nuclei tended to be higher in mule embryos (5.2 ± 0.82) than in horse embryos (3.3 ± 0.3) (P = 0.08). The fragmented nuclei index was similar between mule (0.25 ± 0.1%) and horse (0.22 ± 0.1%) embryos (P = 0.4); the mitotic nuclei index was higher in mule embryos (3.2 ± 0.4%) than in horse embryos (2.2 ± 0.2%) (P = 0.02). In conclusion, embryo morphology of mares bred to a donkey and a horse shares similar nuclear ultrastructure features, except that mule embryos have a higher mitotic index. • Mule embryos share morphological features of horse embryos. • A higher number of mitotic nuclei is displayed in mule embryos. • The horse embryo grading system could apply to mule embryos. [ABSTRACT FROM AUTHOR]

Ribosome inactivating proteins (RIPs) are specific N-β-glycosylases that are well-characterized in plants. Their enzymatic action is to damage ribosomes, thereby blocking protein translation. Recently, several research groups have been working on the screening for these toxins in edible plants to facilitate the use of RIPs as biotechnological tools and biopesticides and to overcome public prejudice. Here, four novel monomeric (type 1) RIPs have been isolated from the seeds of Atriplex hortensis L. var. rubra, which is commonly known as edible red mountain spinach. These enzymes, named hortensins 1, 2, 4, and 5, are able to release the β-fragment and, like many other RIPs, adenines from salmon sperm DNA, thus, acting as polynucleotide:adenosine glycosidases. Structurally, hortensins have a different molecular weight and are purified with different yields (hortensin 1, ~29.5 kDa, 0.28 mg per 100 g; hortensin 2, ~29 kDa, 0.29 mg per 100 g; hortensin 4, ~28.5 kDa, 0.71 mg per 100 g; and hortensin 5, ~30 kDa, 0.65 mg per 100 g); only hortensins 2 and 4 are glycosylated. Furthermore, the major isoforms (hortensins 4 and 5) are cytotoxic toward human continuous glioblastoma U87MG cell line. In addition, the morphological change in U87MG cells in the presence of these toxins is indicative of cell death triggered by the apoptotic pathway, as revealed by nuclear DNA fragmentation (TUNEL assay). [ABSTRACT FROM AUTHOR]

The surge in pharmaceutical consumption, particularly non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen, has raised concerns about their presence in aquatic ecosystems. This study investigated the potential ecological impact of ibuprofen, focusing on the ultrastructure of storage cells in the tardigrade Paramacrobiotus experimentalis, renowned for its resilience to environmental stressors. Individuals were exposed to three ibuprofen concentrations (0.1 μg/L, 16.8 μg/L, and 1 mg/L) over 7 and 28 days. Storage cells were examined using light microscopy, transmission electron microscopy, and confocal microscopy. This study also explored ibuprofen's impact on the process of anhydrobiosis. In the short-term experiment, no ultrastructural changes in tardigrade storage cells were observed across ibuprofen concentrations. However, in the long-term incubation, autophagic structures in storage cell cytoplasm were identified, indicating potential adaptive responses. Individual mitochondria exhibited degeneration, and the rough endoplasmic reticulum displayed slight swelling. No evidence of increased oxidative stress or nuclear DNA fragmentation was observed in any research group. This study elucidates the complex responses of tardigrade storage cells to ibuprofen exposure. The findings emphasize the importance of understanding pharmaceutical impacts on aquatic organisms, highlighting the resilience of tardigrades to specific environmental stressors. [ABSTRACT FROM AUTHOR]

This study was performed to assess the antiapoptotic effect of canine platelet-rich plasma (PRP) treated on the canine adipose-derived mesenchymal stem cells (cMSCs) under cold ischemic conditions. The effect of preventing apoptosis of cMSCs was evaluated in the apoptotic condition induced by cold ischemic injury in vitro. To determine the progression of apoptosis, the changes in cell nucleus were observed using 4’,6-diamidino-2-phenylindole (DAPI) fluorescence staining. In addition, we examined the mitochondrial membrane potential (MMP) and caspase-3 activity. When the cold hypoxic injury was applied to cMSCs, the apoptotic change was observed by DAPI staining, mitochondrial staining for MMP, and caspase-3 assay. PRP significantly decreased the number of apoptotic cells. Nuclear shrinkage and fragmentation of apoptotic cells in control groups were observed by DAPI staining. The MMP was recovered by the treatment of PRP. In addition, when the luminescence intensity was measured for caspase-3 activity, the value was significantly higher in the PRP treated groups than the control groups. The results of this study showed that the PRP may have a beneficial effect on apoptosis induced by cold ischemic injury. [ABSTRACT FROM AUTHOR]

Exposure to time‐varying electromagnetic fields (EMF) has the capacity to influence biological systems. Our results demonstrate that exposure to time‐varying EMF modeled after the physiological firing frequency of intercellular calcium waves can inhibit proliferation and induce apoptosis in malignant cells. Single exposure of B16‐BL6 cells to a Ca2+ EMF for 40 min reduced the number of viable cells by 50.3%. Cell imaging with acridine orange and ethidium bromide dye revealed substantial cellular apoptosis, preapoptotic cells, nuclear fragmentation, and large spacing between cells in the Ca2+ EMF condition when compared to the control condition. The ability of Ca2+ EMF to influence the proliferation and survival of malignant cells suggests that exposure to specific EMF may function as a potential anticancer therapy. [ABSTRACT FROM AUTHOR]

The characterization of wildland firefighters' occupational exposure must consider different exposures, including those at the fire station. The present study aimed to characterize the occupational exposure of 172 Northern Portuguese wildland firefighters in fire stations during the pre-wildfire season of 2021. The biological impact of estimated inhaled doses of PM10 and PM2.5 (indoor/outdoor) was accessed through a buccal micronucleus cytome (BMCyt) assay in exfoliated buccal cells of a subgroup of 80 firefighters. No significant association was found between estimated inhaled doses of PM10 and PM2.5 (mean 1.73 ± 0.43 µg kg−1 and 0.53 ± 0.21 µg kg−1, respectively) and biological endpoints. However, increased frequencies of cell death parameters were found among subjects of the Permanent Intervention Teams (full-time firefighters). The intake of nutritional supplements was associated with a significant decrease in micronucleus frequencies (i.e., DNA damage or chromosome breakage). In addition, our findings showed a significantly increased frequency of cell death endpoints (i.e., nuclear fragmentation) with coffee consumption, while daily consumption of vegetables significantly decreased it (i.e., nuclear shrinkage). Our results provide data on the occupational exposure of wildland firefighters while working in fire stations during the pre-wildfire season, providing the essential baseline for further studies throughout the wildfire season. [ABSTRACT FROM AUTHOR]

Introduction: Charged Particle Therapy plays a key role in the treatment of deep-seated tumours, because of the advantageous energy deposition culminating in the Bragg peak. However, knowledge of the dose delivered in the entrance channel is limited by the lack of data on the beam and fragmentation of the target. Methods: The FOOT experiment has been designed to measure the cross sections of the nuclear fragmentation of projectile and target with two different detectors: an electronic setup for the identification of Z > 3 fragments and a nuclear emulsion spectrometer for Z < 3 fragments. In this paper, we analyze the data taken by exposing four nuclear emulsion spectrometers, with C and C2H4 targets, to 200 MeV/n and 400 MeV/n oxygen beams at GSI Helmholtzzentrum für Schwerionenforschung (Darmstadt, Germany), and we report the charge identification of produced fragments based on the controlled fading induced on nuclear emulsion films. Results: The goal of identifying fragments as heavy as lithium has been achieved. Discussion: The results will contribute to a better understanding of the nuclear fragmentation process in charged particle therapy and have implications for refining treatment planning in the presence of deep-seated tumors. [ABSTRACT FROM AUTHOR]

At the molecular level, several developmental signaling pathways, such as Wnt/β-catenin, have been associated with the initiation and subsequent progression of prostate carcinomas. The present report elucidated the anti-cancerous attributes of an anthraquinone, aloe-emodin (AE), against androgen-independent human prostate cancer DU145 cells. The cytotoxicity profiling of AE showed that it exerted significant cytotoxic effects and increased lactose dehydrogenase levels in DU145 cells (p < 0.01 and p < 0.001). AE also induced considerable reactive oxygen species (ROS)-mediated oxidative stress, which escalated at higher AE concentrations of 20 and 25 µM. AE also efficiently instigated nuclear fragmentation and condensation concomitantly, followed by the activation of caspase-3 and -9 within DU145 cells. AE further reduced the viability of mitochondria with increased cytosolic cytochrome-c levels (p < 0.01 and p < 0.001) in DU145 cells. Importantly, AE exposure was also correlated with reduced Wnt2 and β-catenin mRNA levels along with their target genes, including cyclin D1 and c-myc. Furthermore, the molecular mechanism of AE was evaluated by performing molecular docking studies with Wnt2 and β-catenin. Evidently, AE exhibited good binding energy scores toward Wnt2 and β-catenin comparable with their respective standards, CCT036477 (Wnt2 inhibitor) and FH535 (β-catenin inhibitor). Thus, it may be considered that AE was competent in exerting anti-growth effects against DU145 androgen-independent prostate cancer cells plausibly by modulating the expression of Wnt/β-catenin signaling. [ABSTRACT FROM AUTHOR]

• The acetone extract of Momordica balsamina induces apoptosis in MCF-7 cells. • The apoptosis induced by the extract was attributed to the extract's ability to modulate various apoptosis regulatory proteins. • Treatment of MCF-7 cells with the extract reduced cell invasiveness and migration through the regulation of protein involved in cell invasion and migration. Despite its well-documented nutritional and medicinal properties, Momordica balsamina has not received enough attention in cancer research. As a result, this study investigated whether a crude acetone leaf extract of Momordica balsamina could induce apoptosis and inhibit the migration and invasiveness of MCF-7 breast cancer cells. The MTT assay showed a significant decrease in cell viability after 48 h of treatment with concentrations from 100 – 200 µg/ml of the extract. Light microscopy, the acridine orange/ethidium bromide dual staining assay, and the Muse™ MitoPotential kit showed that the decrease in cell viability is associated with apoptotic cell death. Indeed, extract-treated cells showed signs of chromatin condensation, nuclear fragmentation, and loss of membrane permeability and mitochondrial membrane potential. The scratch wound-healing assay and transwell cell invasion kit showed that the extract inhibited the invasive and migratory abilities of MCF-7 cells, respectively. Also, the extract modulated the expression of proteins involved in the intrinsic and extrinsic pathways of apoptosis, as well as those involved in the regulation of cell invasiveness and migration. Based on these findings, we concluded that Momordica balsamina is a potential source of anticancer compounds. [ABSTRACT FROM AUTHOR]

The main goals of the FragmentatiOn Of Target experiment are the measurements of double-differential fragmentation cross-sections of light particles (Z < 10) in the energy range between 100-800 MeV/u, which is of interest for hadron therapy and space radioprotection applications. The results will fill the gap in experimental data in this range, which is important for improving the treatment planning systems in particle therapy and for enhancing the reliability of risk-assessment models for space radioprotection. Two different setups were designed for this scope, and data have already been taken in several campaigns. In this paper, an overview of the apparatus is described, focusing on the preliminary results of crosssection measurements obtained with a 16O beam at the GSI facility. [ABSTRACT FROM AUTHOR]

In the present study, the shade-dried and powdered leaves of Physalis minima Linn. were subjected to Soxhlet extraction with methanol, concentrated, and further partitioned with n-hexane, chloroform, and ethanol. The in-vitro anti-oxidant activity was evaluated by DPPH radical scavenging assay and the cytotoxic potential by MTT assay in HeLa cell lines. The results of the study revealed that among the three fractions, the chloroform fraction of the methanolic extract showed better antioxidant activity (IC50 = 38.6 µg/mL) compared to standard Ascorbic acid (IC50 = 42.6 µg/mL). Moreover, the results of the MTT assay carried out in HeLa cell lines showed that the chloroform fraction exhibited a promising cytotoxic activity (IC50 = 59.66 µg/mL). Further, the chloroform fraction when subjected to apoptosis assay using acridine orange -ethidium bromide dual staining displayed the morphological characteristics of apoptotic event including nuclear fragmentation and chromatin condensation with orange red and reddish nuclei. The results of our study support the protective effect of chloroform fraction of methanolic extract of leaves of Physalis minima Linn., in the treatment of cervical cancer. [ABSTRACT FROM AUTHOR]

The biological activity of Galium verum herba was exerted on various tumor cell lines with incredible results, but their potential effect on malignant melanoma has not been established yet. Therefore, the current study was structured in two directions: (i) the investigation of the phytochemical profile of diethyl ether (GvDEE) and butanol (GvBuOH) extracts of G. verum L. and (ii) the evaluation of their biological profile on A375 human malignant melanoma cell line. The GvDEE extract showed an FT-IR profile different from the butanol one, with high antioxidant capacity (EC50 of GvDEE = 0.12 ± 0.03 mg/mL > EC50 of GvBuOH = 0.18 ± 0.05 mg/mL). The GvDEE extract also showed antimicrobial potential, especially against Gram-positive bacteria strains, compared to the butanol extract, which has no antimicrobial activity against any bacterial strain tested. The results regarding the antitumor potential showed that both extracts decreased A375 cell viability largely (69% at a dose of 55 µg/mL of the GvDEE extract). Moreover, both extracts induce nuclear fragmentation by forming apoptotic bodies and slight chromatin condensation, which is more intense for GvDEE. Considering the results, one can state that the Galium verum herba possesses antitumor effects on the A375 human malignant melanoma cell line, a promising phytocompound for the antitumor approach to skin cancer. [ABSTRACT FROM AUTHOR]

In the present study, in vitro anti-cancer activity of aerial parts of Epipremnum aureum extracts was performed using the MCF-7 breast cancer cell line. Soxhlet method was used with different solvents for extract preparation. The amount of apoptosis in MCF-7 cells was assessed using flow cytometry for each of the extracts. The chloroform and ethanol extracts had a considerable cytotoxic effect with IC50 values of 32.9 and 45.8 μg/mL respectively, while the conventional medication 5-fluorouracil produced an IC50 value of 19.2 μg/ mL. The microscopic examination of the chloroform and ethanol extracts of E. aureum revealed the presence of apoptotic bodies, nuclear fragmentation, and tiny nuclei with strong chromatin condensation. These results suggest that chloroform extract of E. aureum is more effective against breast cancer. [ABSTRACT FROM AUTHOR]

Background/Aims: Bladder cancer is considered one of the most aggressive neoplasms due to its recurrence and progression profile, and even with the improvement in diagnosis and treatment methods, the mortality rate has not shown a declining trend in recent decades. From this perspective, the search and development of more effective and safer therapeutic alternatives are necessary. Phytochemicals are excellent sources of active principles with therapeutic potential. [6]-Shogaol is a phenolic compound extracted from the ginger rhizomes that has shown antitumor effects in a wide variety of cancer models. However, there is no record in the literature of studies reporting these effects in models of bladder cancer. Thus, this study aimed to investigate the in vitro cytotoxic and pro-apoptotic potential of [6]-Shogaol against murine bladder cancer urothelial cells (MB49). Methods: The cytotoxic effects of [6]-Shogaol on cell viability (MTT method), cell morphology (light microscopy), alteration of proliferative processes (clonogenic assay), oxidative stress pathway (levels of reactive oxygen species) and the induction of apoptotic events (flow cytometry and high-resolution epifluorescence imaging) were evaluated in murine urothelial bladder cancer cell lines (MB49), relative to non-tumor murine fibroblasts (L929). Results: The results showed that [6]-Shogaol was able to induce concentration-dependent cytotoxic effects, which compromised cell viability, exhibiting an inhibitory concentration of 50% of cells (IC50) of 146.8 µM for MB49 tumor cells and 236.0 µM for L929 non-tumor fibroblasts. In addition to inhibiting and altering the proliferative processes if colony formation, it presented pro-apoptotic activity identified through a quantitative analysis and the observation of apoptotic phenotypes, events apparently mediated by the induction of nuclear fragmentation. Conclusion: The data presented suggest that [6]-Shogaol has a higher concentration-dependent cytotoxic and apoptosis-inducing potential in MB49 cells than in L929 fibroblasts. These results may contribute to the development of therapeutic alternatives for bladder cancer. [ABSTRACT FROM AUTHOR]

To examine the potential protective impact and underlying mechanism of nerve growth factor on brain tissue in hyperbilirubinemia rats through the downregulation of Fas/Fas ligand expression. Thirty six healthy Sprague-Dawley rats were selected as normal control group, and 0.5 ml saline was injected intraperitoneally. The remaining 24 rats were injected intraperitoneally with 50 μg/g bilirubin to establish the model of hyperbilirubinemia. After the model was established successfully, they were divided into hyperbilirubinemia group and nerve growth factor intervention group. The rats in the nerve growth factor intervention group were intraperitoneally injected with 50 μg/kg/d nerve growth factor once a day for 3 d. Rats in each group were killed and their brain tissues were taken to wait for detection. The changes of pathological morphology and the Fas and Fas ligand were compared between the normal control group at 72 h and the hyperbilirubinemia group at 6 h, 24 h, 72 h and the nerve growth factor intervention group. In hyperbilirubinemia group and nerve growth factor intervention group, the brain tissue of rats had different degrees of neurocyto edema. After 6 h, the brain tissue of both groups had degeneration. After 24 h, the neuron necrosis was obvious. After 72 h, the brain tissue of both groups had glial cell proliferation, nuclear pyknosis and fragmentation. However, the severity of brain tissue in nerve growth factor intervention group was less than that in hyperbilirubinemia group, while that in normal control group was not obvious change. At 6 h, 24 h and 72 h, the Fas in the hyperbilirubinemia group was raised than that of the normal control group; while this in the nerve growth factor intervention group was reduced than that of hyperbilirubinemia. At 6 h, 24 h and 72 h, the Fas ligand in hyperbilirubinemia group was raised than that of normal control group; and it in nerve growth factor intervention group was reduced than that of hyperbilirubinemia group. The expression of Fas/Fas ligand can be downregulated by nerve growth factor, thereby providing protection to the brain tissue of rats with hyperbilirubinemia. [ABSTRACT FROM AUTHOR]