Changes In Nuclear Morphology Research Articles

Apoptosis Induction Associated with Enhanced ER Stress Response and Up-Regulation of c-Jun/p38 MAPK Proteins in Human Cervical Cancer Cells by Colocasia esculenta var. aquatilis Hassk Extract

Colocasia esculenta var. Aquatilis Hassk, elephant ear (CF-EE) has been widely used as traditional food and medicine. It also shows other therapeutic properties, such as antimicrobial and anti-cancer activity. In this study, we aim to investigate the effect of CF-EE extract on apoptosis induction associated with ER stress in cervical cancer HeLa cells. Cell viability was determined by MTT assay. Assessments of nuclear morphological changes, mitochondrial membrane potential, and intracellular reactive oxygen species (ROS) production were conducted by hoeshst33342, JC-1, and DCFH-DA fluorescence staining, respectively. Sub-G1 DNA content was analyzed by flow cytometry, and protein expression was determined by Western blotting. The results demonstrate that CF-EE extract suppressed HeLa cell growth and induced nuclear condensation and apoptotic bodies. There was also a loss of mitochondrial membrane potential and increased apoptosis marker protein expression, including Bax, cleaved-caspase-7, and cleaved-PARP. In addition, the results show that CF-EE extract induced ROS, increased ER stress proteins (GRP78 and CHOP), enhanced p38 and c-Jun phosphorylation, and inhibited Akt expression in HeLa cells. In summary, CF-EE extract induced apoptotic cell death-associated ROS-induced ER stress and the MAPK/AKT signaling pathway. Therefore, CF-EE extract has anticancer therapeutic potential for cervical cancer treatment in the future.

FUCCItrack: An all-in-one software for single cell tracking and cell cycle analysis

Beyond the more conventional single-cell segmentation and tracking, single-cell cycle dynamics is gaining a growing interest in the field of cell biology. Thanks to sophisticated systems, such as the fluorescent ubiquitination-based cell cycle indicator (FUCCI), it is now possible to study cell proliferation, migration, changes in nuclear morphology and single cell cycle dynamics, quantitatively and in real time. In this work, we introduce FUCCItrack, an all-in-one, semi-automated software to segment, track and visualize FUCCI modified cell lines. A user-friendly complete graphical user interface is presented to record and quantitatively analyze both collective cell proliferation as well as single cell information, including migration and changes in nuclear or cell morphology as a function of cell cycle. To enable full control over the analysis, FUCCItrack also contains features for identification of errors and manual corrections.

Unveiling the Biocompatible Properties of Date Palm Tree (Phoenix dactylifera L.) Biomass-Derived Lignin Nanoparticles.

Searching for sustainable, ecofriendly, and renewable precursors for nanostructured material synthesis is a fascinating area pertaining to feasibility in various applications. Especially, lignin-based material preparation is essential for unraveling the usage of lignin by valorization. Hence, we have synthesized lignin nanoparticles (LNPs) using date palm tree (Phoenix dactylifera L.) biomass as a precursor in this investigation. The LNP’s morphological and thermal features were assessed. Moreover, we have evaluated the LNP’s cytocompatibility properties by adopting in vitro approach. The P. dactylifera L. (PD) biomass-derived LNP’s morphological features show a spherical shape with a 10–100 nm diameter. The LNPs have a decreased cell viability of ∼8% at a high concentration exposure to human mesenchymal stem cells (hMSCs) for 48 h. However, the LNPs do not cause any cellular and nuclear morphology changes in hMSCs. The mitochondrial membrane potential assessment results confirm healthy mitochondria with high mitochondrial membrane potential in LNP-treated cells. The intracellular reactive oxygen species (ROS) generation assay results revealed that LNPs do not trigger ROS generation in hMSCs. We examined the upregulation of GSTM3 and GSR genes and the downregulation of SOD1 genes in LNP-treated hMSCs, but no significant changes were observed. Our study concluded that PD biomass-derived LNPs have a good cytocompatibility and an antioxidant property. Thus, they can be applicable for various biological, cosmetic, and environmental applications.

Antitumour Activity of Different Bioactive Compounds from Hemolymph and Mucus of Mollusca against Human Urinary Bladder Cancer Cell Lines

In this study, we isolated different bioactive compounds and fractions from Rapana venosa (RvH), Helix aspersa (HaH) and Helix lucorum (HlH) and then we evaluated their in vitro antitumour activity on CAL-29 and T- 24 cell lines (transitional cell carcinoma). Also, we performed fluorescence staining with propidium iodide to assess nuclear changes caused by molluscan extracts. Briefly, our results have demonstrated antineoplastic activity of: subunit βc-HaH of H. aspersa hemocyanins to CAL-29 cell line, subunit βc-HaH and subunit RvH II of R. venosa hemocyanin against T-24 cells. Also, fractions from R. venosa hemolymph exerted significant cytotoxic activity against CAL- 29 cells. Furthermore, fluorescence microscopy results demonstrated chromatin condensation and nuclear morphological changes which are typical for apoptosis induction. We assume that the observed death of T-24 and CAL-29 cells is due to the interaction between the oligosaccharide structures of the proteins on the tumour cell membranes and the hemolymph from R. venosa, active components isolated from hemolymph of R. venosa, H. aspersa and H. lucorum, as well as H. aspersa mucus.

Mechanics and functional consequences of nuclear deformations.

As the home of cellular genetic information, the nucleus has a critical role in determining cell fate and function in response to various signals and stimuli. In addition to biochemical inputs, the nucleus is constantly exposed to intrinsic and extrinsic mechanical forces that trigger dynamic changes in nuclear structure and morphology. Emerging data suggest that the physical deformation of the nucleus modulates many cellular and nuclear functions. These functions have long been considered to be downstream of cytoplasmic signalling pathways and dictated by gene expression. In this Review, we discuss an emerging perspective on the mechanoregulation of the nucleus that considers the physical connections from chromatin to nuclear lamina and cytoskeletal filaments as a single mechanical unit. We describe key mechanisms of nuclear deformations in time and space and provide a critical review of the structural and functional adaptive responses of the nucleus to deformations. We then consider the contribution of nuclear deformations to the regulation of important cellular functions, including muscle contraction, cell migration and human disease pathogenesis. Collectively, these emerging insights shed new light on the dynamics of nuclear deformations and their roles in cellular mechanobiology.

Modulation of NF-κB and MAPK signalling pathways by hydrolysable tannin fraction from Terminalia chebula fruits contributes to its anti-inflammatory action in RAW 264.7 cells.

Hydrolysable tannin fraction (HTF) derived from Terminalia chebula fruit pericarps was assessed for its anti-inflammatory potential in LPS-induced RAW 264.7 cells. Its molecular mechanism was also established and compared with individual tannins - chebulagic acid (CH) and corilagin (CO). The effect of HTF on LPS-stimulated RAW 264.7 cells was studied by estimating the release of NO, ROS, cytokines and changes in nuclear morphology by DAPI staining. Furthermore, the effect of HTF, CO and CH was compared with the expression of p65, p38 and pERK proteins by immunoblotting and the mRNA transcript level of COX-2, iNOS and TNF-α by quantitative PCR. The in-silico interactions of various hydrolysable tannins present in HTF with molecular targets of inflammation were studied using Maestro software. HTF at the dose levels of 25, 50 and 100 µg/ml was able to decrease the release of NO, ROS and cytokines from LPS-induced RAW 264.7 cells without disturbing the cell nuclear morphology. Investigation of molecular mechanism revealed that inhibition of NF-κB and MAPK signalling pathways was responsible for its anti-inflammatory action. The effect of HTF was higher than the individual tannins CH and CO. HTF can be developed as an effective anti-inflammatory agent.

Development of optimized novel liposome loaded with 6-gingerol and assessment of its therapeutic activity against NSCLC In vitro and In vivo experimental models

6-Gingerol (Gn) is an active compound derived from ginger which possesses various biological activities. The therapeutic applications of Gn are limited due to its hydrophobic nature. To ease its administration, one of the nano-emulsion methods, liposome was selected to encapsulate Gn. Response Surface Methodology (RSM) was used to optimize liposome ratio. 97.2% entrapment efficiency was achieved at the ratio of 1:20:2 (Drug: Lipid: Cholesterol). The optimized liposome attained size below 200 d nm, spherical shape, negative surface charge and showed sustain release upon physical characterization methods such as FESEM, DLS, Zeta potential, Drug release. The signature FTIR peaks of both free Gn and free liposome (FL) were also observed in Lipo-Gn peak. Lipo-Gn showed significant cytotoxic effect on A549 cells (IC50 160.5 ± 0.74 µM/ml) as well as inhibits the cell migration. DAPI staining showed higher apoptotic nuclear morphological change in the cells treated with Lipo-Gn, and also Lipo-Gn increased the apoptotic percentage in A549 as 39.89 and 70.32 for 12 and 24 h respectively which were significantly more than free Gn. Moreover, the formulation of Lipo-Gn showed significant cell cycle arrest at the G2/M phase compared with free Gn (28.9% and 34.9% in Free Gn vs. 42.7% and 50.1% in Lipo -Gn for 12 and 24 h respectively). Lipo-Gn have been assessed in NSCLC induced BALB/c mice and showed significantly improved pharmacological properties compared to those of free Gn. Thus, Lipo-Gn may be considered for its widening applications against lung cancer.

Disclosing the antitumour potential of the marine bromoditerpene sphaerococcenol A on distinct cancer cellular models

Nature has revealed to be a key source of innovative anticancer drugs. This study evaluated the antitumour potential of the marine bromoditerpene sphaerococcenol A on different cancer cellular models. Dose-response analyses (0.1–100 µM; 24 h) were accomplished in eight different tumour cell lines (A549, CACO-2, HCT-15, MCF-7, NCI-H226, PC-3, SH-SY5Y, SK-MEL-28). Deeper studies were conducted on MFC-7 cells, namely, determination of hydrogen peroxide (H2O2) levels and evaluation of apoptosis biomarkers (phosphatidylserine membrane translocation, mitochondrial dysfunction, Caspase-9 activity, and DNA changes). The ability of the compound to induce genotoxicity was verified in L929 fibroblasts. Sphaerococcenol A capacity to impact colorectal-cancer stem cells (CSCs) tumourspheres (HT29, HCT116, SW620) was evaluated by determining tumourspheres viability, number, and area, as well as the proteasome inhibitory activity. Sphaerococcenol A hepatoxicity was studied in AML12 hepatocytes. The compound exhibited cytotoxicity in all malignant cell lines (IC50 ranging from 4.5 to 16.6 µM). MCF-7 cells viability loss was accompanied by H2O2 generation, mitochondrial dysfunction, Caspase-9 activation and DNA nuclear morphology changes. Furthermore, the compound displayed the lowest IC50 on HT29-derived tumourspheres (0.70 µM), followed by HCT116 (1.77 µM) and SW620 (2.74 µM), impacting the HT29 tumoursphere formation by reducing their number and area. Finally, the compound displayed low cytotoxicity on AML12 hepatocytes without genotoxicity. Overall, sphaerococcenol A exhibits broad cytotoxic effects on different tumour cells, increasing H2O2 production and apoptosis. It also affects colorectal CSCs-enriched tumoursphere development. These data highlight the relevance to include sphaerococcenol A in further pharmacological studies aiming cancer treatments.

A neutrophil-based test as an auxiliary tool for substantiating the diagnosis of bovine tuberculosis.

Bovine tuberculosis (bTB) is still a prominent threat to animal health; lacking an efficient vaccine, other than BCG to get rid of tuberculosis, the most effective way for this is culling and slaughtering the infected animals. There are several cellular, serological, and molecular tests for the diagnosis of the disease but the most practical one at the field level is the double skin testing with bovine and aviary tuberculins. This is not a very specific test but is sensitive enough to identify most diseased animals; adjunct practical tests are desirable to strengthen the utility of skin tests. All lymphoid and myeloid cells participate, in diverse grades, in the immune response to tuberculosis with neutrophils playing an unintended pathologic role. The study aimed to investigate the response of neutrophils to agents present in the sera of tuberculous cows. We have developed a neutrophil-based test (N BT) to identify diseased cows within a herd suspected of having tuberculosis; a positive N BT correlates with a positive double skin test. In this test, healthy neutrophils are incubated with the sera of healthy or tuberculous cows for 3 and 6 h, and the nuclear morphologic changes are recorded and analyzed. Sera from tuberculous but not from healthy cows induce nuclear alterations including pyknosis, swelling, apoptosis, and sometimes NETosis, in healthy neutrophils, and CFP 10 and ESAT 6 participate in the phenomenon. We propose the N BT as an auxiliary tool for substantiating the diagnosis of bTB reinforcing the PPD test outcome to help decide whether or not a cow should be sacrificed.

Epigenetic Landscape of HIV-1 Infection in Primary Human Macrophage

Human immunodeficiency virus (HIV)-infected macrophages are long-lived cells that sustain persistent virus expression, which is both a barrier to viral eradication and contributor to neurological complications in patients despite antiretroviral therapy (ART). To better understand the regulation of HIV-1 in macrophages, we compared HIV-infected primary human monocyte-derived macrophages (MDM) to acutely infected primary CD4 T cells and Jurkat cells latently infected with HIV (JLAT 8.4). HIV genomes in MDM were actively transcribed despite enrichment with heterochromatin-associated H3K9me3 across the complete HIV genome in combination with elevated activation marks of H3K9ac and H3K27ac at the long terminal repeat (LTR). Macrophage patterns contrasted with JLAT cells, which showed conventional bivalent H3K4me3/H3K27me3, and acutely infected CD4 T cells, which showed an intermediate epigenotype. 5'-Methylcytosine (5mC) was enriched across the HIV genome in latently infected JLAT cells, while 5'-hydroxymethylcytosine (5hmC) was enriched in CD4 cells and MDMs. HIV infection induced multinucleation of MDMs along with DNA damage-associated p53 phosphorylation, as well as loss of TET2 and the nuclear redistribution of 5-hydoxymethylation. Taken together, our findings suggest that HIV induces a unique macrophage nuclear and transcriptional profile, and viral genomes are maintained in a noncanonical bivalent epigenetic state. IMPORTANCE Macrophages serve as a reservoir for long-term persistence and chronic production of HIV. We found an atypical epigenetic control of HIV in macrophages marked by heterochromatic H3K9me3 despite active viral transcription. HIV infection induced changes in macrophage nuclear morphology and epigenetic regulatory factors. These findings may identify new mechanisms to control chronic HIV expression in infected macrophages.

B Cells Adapt Their Nuclear Morphology to Organize the Immune Synapse and Facilitate Antigen Extraction.

Upon interaction with immobilized antigens, B cells form an immune synapse where actin remodeling and re-positioning of the microtubule-organizing center (MTOC) together with lysosomes can facilitate antigen extraction. B cells have restricted cytoplasmic space, mainly occupied by a large nucleus, yet the role of nuclear morphology in the formation of the immune synapse has not been addressed. Here we show that upon activation, B cells re-orientate and adapt the size of their nuclear groove facing the immune synapse, where the MTOC sits, and lysosomes accumulate. Silencing the nuclear envelope proteins Nesprin-1 and Sun-1 impairs nuclear reorientation towards the synapse and leads to defects in actin organization. Consequently, B cells are unable to internalize the BCR after antigen activation. Nesprin-1 and Sun-1-silenced B cells also fail to accumulate the tethering factor Exo70 at the center of the synaptic membrane and display defective lysosome positioning, impairing efficient antigen extraction at the immune synapse. Thus, changes in nuclear morphology and positioning emerge as critical regulatory steps to coordinate B cell activation.

Nondimensional mechanical parameters for characterising nuclear morphology

Nuclear morphology is an important indicator of cell function. It is regulated by a variety of factors such as the osmotic pressure difference between the nucleoplasm and cytoplasm, cytoskeletal forces, elasticity of the nuclear envelope and chromosomes. Nucleus shape and size are typically quantified using multiple geometrical quantities that are not necessarily independent of one another. This interdependence makes it difficult to decipher the implications of changes in nuclear morphology. We resolved this by analyzing nucleus shapes of populations for multiple cell lines using a mechanics-based model.

Examination ofwhite blood cell indicators for three different ploidy level sturgeon species reared inanindoor recirculation aquaculture system for one year.

Functional diploid Acipenser ruthenus, functional tetraploid Acipenser gueldenstaedtii and functional hexaploid Acipenser brevirostrum juveniles were sampled monthly for one year, and the white blood cell indicators were determined. The total number ofleukocytes (TL) was 40.93± 17.24× 109/l for the diploids, 20.63± 11.20×109/l for the tetraploids, 14.13± 7.72×109/l for the hexaploids. The TL decreased with anincreasing ploidy level. The highest number ofleukocytes was reached during September and October for A.ruthenus and A.brevirostrum, from October toJanuary for A.gueldenstaedtii (astatistically significant finding). The lymphocytes dominated (76.89-80.14%) inthe differential counts and were found tobe reduced inJune and July ineach group. Granulocytes were represented byneutrophils and eosinophils. Counting from all the leukocytes, the neutrophils represented 13.0-18.7% and eosinophils represented 5.7-6.1%. Increasing number ofnuclear segments inthe granulocytes was dependent onthe increasing ploidy level. Nuclear segmentation inthe lymphocytes was acommon finding inhigher ploidy level groups. The data suggest asignificant effect ofploidy level onthe total number ofleukocytes and morphological nuclear changes inthe granulocytes and lymphocytes. The seasonal variation inthe differential leukocyte counts depends onthe species and the influence ofvarious external conditions rather than the ploidy level.

Rapid Prototypable Biomimetic Peristalsis Bioreactor Capable of Concurrent Shear and Multi-Axial Strain

Peristalsis is a nuanced mechanical stimulus comprised of multi-axial strain (radial and axial strain) and shear stress. Forces associated with peristalsis regulate diverse biological functions including digestion, reproductive function, and urine dynamics. Given the central role peristalsis plays in physiology and pathophysiology, we were motivated to design a bioreactor capable of holistically mimicking peristalsis. We engineered a novel rotating screw-drive based design combined with a peristaltic pump, in order to deliver multi-axial strain and concurrent shear stress to a biocompatible polydimethylsiloxane (PDMS) membrane “wall.” Radial indentation and rotation of the screw drive against the wall demonstrated multi-axial strain evaluated via finite element modeling. Experimental measurements of strain using piezoelectric strain resistors were in close alignment with model-predicted values (15.9 ± 4.2% vs. 15.2% predicted). Modeling of shear stress on the “wall” indicated a uniform velocity profile and a moderate shear stress of 0.4 Pa. Human mesenchymal stem cells (hMSCs) seeded on the PDMS “wall” and stimulated with peristalsis demonstrated dramatic changes in actin filament alignment, proliferation, and nuclear morphology compared to static controls, perfusion, or strain, indicating that hMSCs sensed and responded to peristalsis uniquely. Lastly, significant differences were observed in gene expression patterns of calponin, caldesmon, smooth muscle actin, and transgelin, corroborating the propensity of hMSCs toward myogenic differentiation in response to peristalsis. Collectively, our data suggest that the peristalsis bioreactor is capable of generating concurrent multi-axial strain and shear stress on a “wall.” hMSCs experience peristalsis differently than perfusion or strain, resulting in changes in proliferation, actin fiber organization, smooth muscle actin expression, and genetic markers of differentiation. The peristalsis bioreactor device has broad utility in the study of development and disease in several organ systems.

An Imaging and Computational Algorithm for Efficient Identification and Quantification of Neutrophil Extracellular Traps.

Neutrophil extracellular traps (NETs) are associated with multiple disease pathologies including sepsis, asthma, rheumatoid arthritis, cancer, systemic lupus erythematosus, acute respiratory distress syndrome, and COVID-19. NETs, being a disintegrated death form, suffered inconsistency in their identification, nomenclature, and quantifications that hindered therapeutic approaches using NETs as a target. Multiple strategies including microscopy, ELISA, immunoblotting, flow cytometry, and image-stream-based methods have exhibited drawbacks such as being subjective, non-specific, error-prone, and not being high throughput, and thus demand the development of innovative and efficient approaches for their analyses. Here, we established an imaging and computational algorithm using high content screening (HCS)—cellomics platform that aid in easy, rapid, and specific detection as well as analyses of NETs. This method employed membrane-permeable and impermeable DNA dyes in situ to identify NET-forming cells. Automated algorithm-driven single-cell analysis of change in nuclear morphology, increase in nuclear area, and change in intensities provided precise detection of NET-forming cells and eliminated user bias with other cell death modalities. Further combination with Annexin V staining in situ detected specific death pathway, e.g., apoptosis, and thus, discriminated between NETs, apoptosis, and necrosis. Our approach does not utilize fixation and permeabilization steps that disturb NETs, and thus, allows the time-dependent monitoring of NETs. Together, this specific imaging-based high throughput method for NETs analyses may provide a good platform for the discovery of potential inhibitors of NET formation and/or agents to modulate neutrophil death, e.g., NETosis-apoptosis switch, as an alternative strategy to enhance the resolution of inflammation.

Evaluation of Cytotoxic Potential of Digera muricata Leaf Extract on Lung Cancer Cell Line

Lung cancer is the second most frequent cancer, accounting for one out of every five male cancers and one out of every nine female cancers. Treatment for lung cancer is determined by the disease's cell type, the extent to which it has spread, and the patient's overall health. It is common knowledge that tumours impart resistance to chemotherapeutic medicines or radiation in part owing to apoptotic pathway dysfunction in cancer cells. Digera muricata (D.muricata) has been used as medicinal remedies for various ailments due to its antioxidant, anti-inflammatory, anti-bacterial, anti-tumor activity. The objective of the study was to examine the cytotoxic activity of ethanolic leaf extract of D.muricata on lung cancer cell lines. The cytotoxic potency of D.muricata leaf extract was carried out by MTT assay against the lung cancer cell line (A549). Different concentrations of D.muricata ethanolic leaf extract (25-150µg/ml) were treated for 24h. Furthermore, the morphological changes were analysed using phase contrast microscopy. Pro-apoptotic and nuclear morphological changes in D. muricata ethanolic leaf extract treated cells were examined using DAPI staining. The ethanolic leaf extract of D.muricata showed the dose dependent cytotoxic potency against the A549 cell line which confirmed with greater morphological changes upon 24 hrs treatment. The MTT assay clearly showed that the D.muricata treatment has significantly reduced the cell viability when the concentration was increased for 24hrs. We observed IC-50 dose at 50 μg/ml concentration. DAPI staining clearly showed condensed chromatin and fragmented nuclei in treated lung cancer cells. All these results clearly showed that ethanolic extract of D. muricata treatment significantly inhibited the cell proliferation and induced apoptosis in lung cancer cells.

Anticancer effect of Digera muricata Leaf Extract against Prostate Cancer Cell Lines

Background: Prostate cancer has become a major health problem globally during the last few decades. It is the second most frequently diagnosed cancer in men worldwide and the fifth most common cancer overall. Chemotherapy or other cancer treatments including androgen depletion therapy, show high toxicity and cause serious side effects in cancer patients. Thus, there have been many studies conducted to find various natural products as potential anticancer drug candidates with low toxicity and fewer side effects for the treatment and prevention of prostate cancer. The Digera muricata of the genus Digera Forssk and family Amaranthaceae Juss. is a wild edible plant. The presence of phenol, flavonoids, alkaloids, terpenes, sterols, tannins, glycosides and lignins is seen in Digera muricata leaf extract, which are reported to have cytotoxic activities.
 Aim of the Study: The aim of this study is to assess the cytotoxic effects of Digera muricata leaf extract on prostate cancer cell line.
 Materials and Methods: The cytotoxic potency of Digera muricata leaf extract was carried out by MTT (3-[4,5-dimethylthiazol-2yl]-2,5-diphenyl-tetrazolium bromide) assay against the prostate cancer (PC-3) cell line. PC-3 cells were treated with different concentrations of Digera muricata leaf extract (25-150μg/ml) for 24h. Furthermore, the morphological changes were analysed using phase contrast microscopy and nuclear morphological changes examined using DAPI (4′,6-diamidino-2-phenylindole) staining under the fluorescence microscopy
 Results: The MTT assay showed decreased cell vitality with increased concentration of Digera muricata leaf extract. The morphological study showed that the number of cells decreased after treatment and the cells exhibited cell shrinkage and cytoplasmic membrane blebbing. The treated cells also showed condensed chromatin and nuclear fragmentation
 Conclusion: Within the limits of this study it can be demonstrated that the leaf extract of Digera muricata were cytotoxic and induced apoptosis to the cancer cells.

Evaluation of Anticancer Effect of Lippia nodiflora Leaf Extract on Lung Cancer Cells

Background: Lung cancer, also known as bronchogenic carcinoma, is a kind of cancer that begins in the lung parenchyma or inside the bronchi. In the United States, it is one of the top causes of cancer-related fatalities. Lippia nodiflora is a flowering, broadleaf plant native to South America. Lippia nodiflora has been used as a natural medicine for a number of disorders due to its antioxidant, anti-inflammatory, anti-bacterial, and anti-tumor characteristics.
 Aim of the Study: To evaluate the anticancer effect of Lippia nodiflora leaf extract on lung cancer.
 Materials and methods: The cytotoxic effect of Lippia nodiflora leaf extract was assessed by an MTT assay. The A-549 cells were treated with different concentrations of Lippia nodiflora leaf extract (10-120 μg) for 24h. Analysis of changes in the cell morphology is examined by phase contrast microscope. For the nuclear morphological changes analysis, DAPI staining was used in lung cancer cells. The apoptotic nuclei were viewed under a fluorescent microscope.
 Results: In our study, Lippia nodiflora leaf extract treated cells showed significant reduction of cell viability in lung cancer cells. 50% of the inhibition was observed in a 20 μg/ml concentration, which has been taken as the inhibitory concentration (IC-50) which was used for further analyses. The pro-apoptotic effect of Lippia nodiflora leaf extract in A549 cell line was determined by DAPI staining under a fluorescence microscope. Lippia nodiflora leaf extract treatment alters the nuclear morphology and condensed nuclei in lung cancer cells at 20μg/ml concentration.
 Conclusion: Overall, it can be concluded that Lippia nodiflora presents anticancer activity at 20μg/ml concentration at 24 hrs againstthe lung cancer cell line. However, further research is warrented to find out the molecular mechanism of cytotoxicity of effect this plant extract against lung cancer.

Loss of lamin-B1 and defective nuclear morphology are hallmarks of astrocyte senescence in vitro and in the aging human hippocampus.

The increase in senescent cells in tissues, including the brain, is a general feature of normal aging and age‐related pathologies. Senescent cells exhibit a specific phenotype, which includes an altered nuclear morphology and transcriptomic changes. Astrocytes undergo senescence in vitro and in age‐associated neurodegenerative diseases, but little is known about whether this process also occurs in physiological aging, as well as its functional implication. Here, we investigated astrocyte senescence in vitro, in old mouse brains, and in post‐mortem human brain tissue of elderly. We identified a significant loss of lamin‐B1, a major component of the nuclear lamina, as a hallmark of senescent astrocytes. We showed a severe reduction of lamin‐B1 in the dentate gyrus of aged mice, including in hippocampal astrocytes, and in the granular cell layer of the hippocampus of post‐mortem human tissue from non‐demented elderly. The lamin‐B1 reduction was associated with nuclear deformations, represented by an increased incidence of invaginated nuclei and loss of nuclear circularity in senescent astrocytes in vitro and in the aging human hippocampus. We also found differences in lamin‐B1 levels and astrocyte nuclear morphology between the granular cell layer and polymorphic layer in the elderly human hippocampus, suggesting an intra‐regional‐dependent aging response of human astrocytes. Moreover, we described senescence‐associated impaired neuritogenic and synaptogenic capacity of mouse astrocytes. Our findings show that reduction of lamin‐B1 is a conserved feature of hippocampal cells aging, including astrocytes, and shed light on significant defects in nuclear lamina structure which may contribute to astrocyte dysfunctions during aging.

Chemoprevention of NCI-H322 Lung Cancer Cells Proliferation and Tumor Regression in Murine Ascites Model by Dietary Flavonoid Quercetin

Prunus cerasus L (Sour cherries) contain diverse secondary metabolites which exhibit various biological activities, including anticancer, antioxidant, and anti-inflammatory properties. The present study aimed to determine the anticancer efficacy of four compounds, quercetin, daidzin, rutin, and chlorogenic acid, isolated from Prunus cerasus fruit. The antiproliferative activity of four cherry isolates was determined against five different cancer cell lines (NCI-H322, A549, THP-1, MCF-7, and PC-3) by Tetrazolium bromide assay, followed by apoptosis Cell cycle analyses, mitochondrial membrane potential, cell migration test, and in vivo Ehrlich Ascites Carcinoma studies using potent bioactive lead. The cytotoxicity profile of the four molecules demonstrated that quercetin induced significant cell growth inhibition in all cancer cell lines with paramount 79% cytotoxicity against NCI-H322 lung cancer cells (IC50 value 24μM). Incubation of NCI-H322 cells with quercetin showed a concentration-dependent increase in hypo-diploid sub G0/G1 DNA fraction, exhibited consequential changes in nuclear morphology, and caused mitochondrial transmembrane potential loss of 60.3% augmented at 30 µM. Pertaining to in vivo potency, quercetin manifested 89% tumor inhibition at 50 mg/kg body weight in EAC-bearing mice. The current studies raise the potential usefulness of quercetin in chemoprevention against lung cancer cells and support its empirical use as a promising nutraceutical agent.