Percentage Of Necrotic Cells Research Articles

Seminal Plasma-Derived Exosome Preserves the Quality Parameters of the Post-Thaw Semen of Bulls with Low Freezeability.

Introduction: Sperm cryopreservation is a useful storage technique in artificial insemination. Nanoparticles and nanovesicles such as exosomes are widely used in sperm cryopreservation procedures to alleviate cold-induced injury inflicted during sperm freezing. Objective: The objective of the present study was to examine the impact of varying concentrations of exosomes derived from seminal plasma added to a freezing extender on the quality of post-thawed bull sperm. Methods: Five Holstein bulls were chosen based on their samples having less than 30% progressive motility. After exosome extraction, semen samples from bulls (n = 5) with progressive sperm motility ≤30% were collected, diluted with different exosome concentrations (0, 25, 50, and 100 μg/mL), and aspirated into 0.5 mL straws. After the freeze-thaw process, sperm total and progressive motility, viability, morphology, plasma membrane integrity, mitochondrial activity, and apoptosis status were assessed. Furthermore, the expression levels of annexin (ANX1), dystrophy-associated Fer-1-like protein (DYSF), fibronectin 1 (FN1), and reactive oxygen species modulator 1 (ROMO1) were evaluated via real-time polymerase chain reaction (PCR). Results: Adding different concentrations of exosomes (25, 50, and 150 μg/mL) significantly increased the progressive motility, viability, and membrane integrity of sperm compared with the control group (p < 0.05). For the apoptosis index, treatment with 100 μg/mL exosomes significantly increased the percentage of live cells (p < 0.05), while the percentage of necrotic cells decreased significantly (p < 0.05) compared with 25 μg/mL exosome. The results of quantitative PCR showed that the expression levels of ANX1 were significantly (p < 0.05) upregulated at 50 μg/mL exosome, and the expression of ROMO1, FN1, and DYSF were downregulated upon treatment with different exosome concentrations. Conclusions: In conclusion, supplementing the freezing diluent with exosome-derived seminal plasma could preserve the quality parameters of the post-thaw semen of the bull with low freezeability and could be used as a helpful method for reproductive programs.

VIABILITY OF BLOOD LEUKOCYTES OF RATS AFTER IMPLANTATION OF POLYPROPYLENE SURGICAL MESH WITH A TANTALUM-BASED COATING AND ITS DERIVATIVES

Aim. To investigate the viability and types of cell death of peripheral blood leukocytes in experimental animals after implantation of surgical meshes with a tantalum-based coating and its derivatives. Materials and methods. The experimental group included 40 male rats of the WAG population. Polypropylene surgical mesh was surgically implanted between the abdominal wall and sections of the large intestine with different types of coatings. After 28 days, collected blood was analyzed by a BD FACSCanto™ II flow cytometer. Results. It was determined that there were no significant changes in the viability of blood leukocytes between the animals of the intact group and the group of animals that underwent surgery without implantation. Analysis of leukocyte viability in groups of rats implanted with tantalum and tantalum oxide-coated meshes showed a slight decrease in viable cells compared with the results of the intact group. At the same time, the percentage of necrotic cells showed a slight increase. In the group of rats implanted with a mesh with tantalum nitride-coated, a decrease in viable leukocytes was determined in comparison with the results of the intact group by 12,9%, while the percentage of necrotic leukocytes was 3,8% higher. A 16,2% decrease in viable leukocytes was determined in the group of rats implanted with a non-coated mesh compared to the results of the intact group. At the same time, the percentage of necrotic cells was 6,9% higher. Conclusions. Implantation of uncoated and tantalum nitride-coated surgical meshes was found to decrease the percentage of viable blood leukocytes in rats compared to intact animals, while implantation of tantalum- and tantalum-oxide-coated surgical meshes did not significantly decrease viable white blood cells. leukocytes.

P97 inhibitor promotes apoptosis on colon cancer cells

Background: p97 is a multifunctional protein that plays an important role in DNA damage repair and processes related to programmed cell death. We used CB-5083, a specific p97 inhibitor, to evaluate the effect of inhibiting cluster proliferation and promoting apoptosis on HCT116 colon cancer cells. Materials and methods: Experimental research method used p97 inhibitor CB-5083 (Sellechem) and colon cancer cell line HCT116 (code: CCL-247, ATCC). Colony formation assay and apoptotic assay were used in this study. Data were analysed on ImageJ and GraphPad Prism 9 software. Results: The density and size of cell colonies in the 0.25 µM CB5083 treatment group were statistically significantly smaller than the other groups. The 0.25 µM treatment group had a statistically significant higher percentage of apoptotic cells compared to the 0,125µM group and the control group (p &lt; 0.0001). The percentage of necrotic cells in the 0.125 µM and 0.25 µM treatment groups was statistically significantly higher than the control group (p &lt; 0.0001). Conclusion: Inhibiting p97 by CB5083 suppressed cell colony formation and promoted apoptosis on HCT116 colon cancer cell line. Key words: p97, DNA damage repair, colon cancer, apoptosis.

Ameliorative effect of zinc oxide-chitosan conjugates on the anticancer activity of cisplatin: Approach for breast cancer treatment

Breast cancer is the second most prevalent cancer affecting both males and females, comprising nearly 30 % of all cancer cases. While chemotherapeutic agents, such as cisplatin (Cis), have proven successful in cancer treatment, concerns persist regarding their efficacy and the potentially dangerous side effects. Consequently, there is a crucial and ongoing need to develop approaches that minimize side effects associated with chemotherapy. In the present work, various types of nanoparticles (NPs) were synthesized and loaded with Cis. Cis was conjugated with nanocarriers such as zinc oxide (ZnO), ZnO modified with mandelic acid and graphene oxide (GO), chitosan (CS), and CS modified with ZnO and GO to enhance the selectivity of Cis towards cancer cells. Zeta potentials and particles size were assessed using electrophoretic light scattering and dynamic light scattering. NPs were characterized using transmission electron microscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction. The impact of standalone Cis as well as its nanoconjugated form on the behavior of MCF-7 cell line was investigated using WST-1 cell proliferation and apoptosis/necrosis assays. Experimental findings revealed that among the various NPs tested, ZnO, and CS NPs exhibited the highest loading percentage of Cis, surpassing the loading percentages achieved with other NPs. Cytotoxicity assay showed the enhanced effect of Cis when conjugated with ZnO and CS NPs. Flow cytometry-based assays and confocal microscopy confirmed that ZnO/Cis and CS/Cis induced apoptosis. The cisplatin-nanocomplex exhibited a descending order of early apoptosis and late apoptosis in the following order: ZnO, Cis, CS, ZnO-M, CS-GO, ZnO-GO, CS-ZnO, and CS-ZnO, Cis, CS, CS-GO, ZnO-M, ZnO, ZnO-GO, respectively. None of the nanoparticle complexes displayed a significant percentage of necrotic cells, with the highest percentage reaching 4.65 % in the case of CS-GO/Cis.

Lysicamine Reduces Protein Kinase B (AKT) Activation and Promotes Necrosis in Anaplastic Thyroid Cancer.

Anaplastic thyroid cancer (ATC) is an aggressive form of thyroid cancer (TC), accounting for 50% of total TC-related deaths. Although therapeutic approaches against TC have improved in recent years, the survival rate remains low, and severe adverse effects are commonly reported. However, unexplored alternatives based on natural compounds, such as lysicamine, an alkaloid found in plants with established cytotoxicity against breast and liver cancers, offer promise. Therefore, this study aimed to explore the antineoplastic effects of lysicamine in papillary TC (BCPAP) and ATC (HTH83 and KTC-2) cells. Lysicamine treatment reduced cell viability, motility, colony formation, and AKT activation while increasing the percentage of necrotic cells. The absence of caspase activity confirmed apoptosis-independent cell death. Necrostatin-1 (NEC-1)-mediated necrosome inhibition reduced lysicamine-induced necrosis in KTC-2, suggesting necroptosis induction via a reactive oxygen species (ROS)-independent mechanism. Additionally, in silico analysis predicted lysicamine target proteins, particularly those related to MAPK and TGF-β signaling. Our study demonstrated lysicamine's potential as an antineoplastic compound in ATC cells with a proposed mechanism related to inhibiting AKT activation and inducing cell death.

Analysis of the antimelanogenic activity of zinc and selenium in vitro.

Melasma is an acquired chronic condition characterized by hyperchromic patches in photo-exposed areas. The search for new compounds for the treatment of melasma without side effects is constant. In this context, the aim of this study was to investigate the in vitro cytotoxic and antimelanogenic effects of the trace elements Zinc (Zn) and Selenium (Se). In this study, we evaluated the effects of 30µM hydroquinone, this concentration did not alter mitochondrial function (MTT assay), but increased the percentage of necrotic cells and levels of reactive species. Furthermore, it showed no influence on tyrosinase activity and melanin content. Unlike hydroquinone, exposure for 48h to 100µM Zn and 1 and 5µM Se had no significant influence on the analysis of reactive species, as well as on the percentage of necrotic cells. Still, specifically in relation to 100µM Zn, it decreased the melanin content. Given the above, the trace elements Zn and Se did not show toxicity at the concentrations tested and Zn showed a promising effect, however, the mechanism needs to be better explored in order to contribute to new and updated research in the fight against melasma with a perspective of therapeutic use.

HWJMSCs inhibit inflammation and apoptosis in an ARDS cell model

Acute respiratory distress syndrome (ARDS) is a type of lung failure caused by fluids and hypoxemia. Mesenchymal stem cells (MSCs) have been shown to decrease levels of pro-inflammatory mediators and inflammatory cells. These cells have anti-inflammatory, anti-apoptotic, and anti-microbial activity, and protect against lung injury. ObjectiveThis research evaluated the potential of human Wharton's jelly MSCs (hWJMSCs) to inhibit inflammation and apoptosis in lipopolysaccharide (LPS)-induced rat lung cells (L2). MethodshWJMSC treatment in LPS-induced rat lung cells was performed with 1:1, 1:5, 1:10, or 1:25 ratios of hWJMSCs to L2 cells. The gene expression of angiotensin-converting enzyme-2 (ACE-2), receptor for advanced glycation end products (RAGE), nuclear factor kappa B (NFκB), and C-X-C motif chemokine ligand-9 (CXCL-9) was quantified with RT-PCR, and the levels of C-reactive protein (CRP), interleukin-12 (IL-12), and tumor necrosis factor-alpha (TNF-α) were measured with ELISA. ResultshWJMSCs increased ACE-2 gene expression, and decreased CXCL-9, NFκB, and RAGE gene expression. The treatment also suppressed CRP, TNF-α, and IL-12 levels, and increased the percentage of live cells, but decreased the percentages of necrotic cells and apoptotic cells in inflammatory rat lung cells, which served as an ARDS cell model. ConclusionCo-culture of hWJMSCs and L2 cells mitigated inflammation through increasing ACE-2 gene expression, and decreasing CXCL-9, NFκB, and RAGE gene expression; decreasing TNF-α and CRP protein levels; and decreasing necrosis, and early and late apoptosis. A co-culture ratio of 1:1 was most effective.

Noni simplistic effect with Chicken Shank Gelatin Film on White Rat Spleen Exposed to Dexamethasone

Dexamethasone is a corticosteroid drug belong to glucocorticoid group. Dexamethasone is immunosuppressant and anti-inflammatory in various inflammatory conditions. Side effects of its use can cause cell apoptosis in various organs such as the spleen. The immunosuppressant effect of dexamethasone can reduce and inhibit peripheral lymphocytes and macrophages until the death of lymphoid cells in the white pulps of the spleen. The simultaneous effect of administering chicken shank gelatin and noni has the potential to improve the structure of the spleen. This study was aimed to prove and obtain effective and safe dose of chicken shank gelatin and noni on the spleens of rats exposed to dexamethasone. The research was carried out experimentally in the laboratory with a completely randomized design (CRD). A total of 25 heads male rats were grouped into five treatments and each treatment consist of five repetitions. There was a treatment group P1 as a negative control, P2 as a positive control (dexamethasone 5 mg/kg BW), P3-P5 (dexamethasone 5 mg/kg + gelatin 1.585 mg/kg + noni simplicia 50; 112; 250 mg/kg BW). The results of the study showed an increase in the area of white pulps and a decrease in the percentage of necrotic cells in the spleen, however, it did not increase the relative weight of the spleen and serum albumin levels (P&gt;0.05). In conclusion, the effective and safe dose for the spleen organs of rats exposed to dexamethasone is 250 mg/kg BW.

The effect of cerium dioxide nanoparticles on the viability of hippocampal neurons in Alzheimer's disease modeling.

The possibilities of using nanoparticle materials based on cerium dioxide (CNPs) are exciting since they are low toxic and have specific redox, antiradical properties. It can be supposed that CNPs' biomedical use is also relevant in neurodegenerative diseases, especially Alzheimer's disease (AD). AD is known as the pathologies leading to progressive dementia in the elderly. The factor that provokes nerve cell death and cognitive impairment in AD is the pathological accumulation of beta-amyloid peptide (Aβ) in the brain tissue. In our studies, we examined the impact of Aβ 1-42 on neuronal death and evaluated the potential neuroprotective properties of CNPs during AD modeling in cell culture. Our findings show that, under AD modeling conditions, the number of necrotic neurons increased from 9.4% in the control to 42.7% when Aβ 1-42 was used. In contrast, CNPs alone showed low toxicity, with no significant increase in the number of necrotic cells compared to control conditions. We further explored the potential of CNPs as a neuroprotective agent against Aβ-induced neuronal death. We found that introducing CNPs 24 h after Aβ 1-42 incubation or prophylactically incubating hippocampal cells with CNPs 24 h before amyloid administration significantly reduced the percentage of necrotic cells to 17.8 and 13.3%, respectively. Our results suggest that CNPs in the cultural media can significantly reduce the number of dead hippocampal neurons in the presence of Aβ, highlighting their neuroprotective properties. These findings suggest that CNPs may hold promise for developing new treatments for AD based on their neuroprotective properties.

Renal tubular epithelial cell necroptosis promotes tubulointerstitial fibrosis in patients with chronic kidney disease.

Renal fibrosis, a common pathological manifestation of virtually all types of chronic kidney disease (CKD), ultimately predisposes patients to end-stage renal disease. However, there is no effective therapy for renal fibrosis. Our earlier studies proved that RIP3-mediated necroptosis might be an important mode of renal tubular cell death in rats with chronic renal injury. Under transmission electron microscopy (TEM), we found morphological changes in the necrosis of human renal tissue, and the percentage of necrotic cells increased significantly in patients with stages 2 and 3a CKD. Immunofluorescence analyses showed that the percentages of TUNEL+ /RIP3+ double-positive and TUNEL+ /MLKL+ double-positive tubular epithelial cells in renal tubules of patients with stages 2 and 3a CKD were significantly increased compared to those in control patients without renal disease. Immunohistochemistry analyses of renal biopsy specimens from patients with CKD revealed RIP3, MLKL, and p-MLKL upregulation in patients with stages 2 and 3a CKD, suggesting that necroptosis of renal tubular epithelial cells in CKD patients occurs, and the peak of necroptosis was in stages 2 and 3a CKD. We showed that profibrotic factor proteins (TGF-β1, Smad2 and Smad3) and fibroblast activation markers (α-SMA and Vimentin) were specifically upregulated in stage 2 and 3a CKD patients. In addition, Pearson correlation analysis showed that the percentage of necroptotic renal tubular epithelial cells was positively correlated with TGF-β1 and collagen-I. We also showed that RIP1/3 or MLKL inhibitors decreased the expression of RIP3, MLKL, TGF-β1, and Smad3 in HK-2 cells treated with TNF-α. FGF-2, α-SMA, Vimentin and FN were overexpressed in the hRIFs cultured with the supernatant of necroptotic HK-2 cells, whereas necroptosis blockers (Nec-1s, GSK'872 and NSA) and TGF-β1/Smad3 pathway antagonists (LY364947 and SIS3) reduced FGF-2, α-SMA, Vimentinand FN levels. Collectively, necroptosis of renal tubular epithelial cells in CKD patients occurs, and the peak of necroptosis was in stages 2 and 3a CKD. Renal tubular epithelial cell necroptosis mediates renal tubulointerstitial fibrosis in patients with chronic kidney disease, which is related to the TGF-β1/Smad3 signaling pathway.

Suppression Effects of Excessively Expressed Gene BCL-2 in Cell Lines of Prostate Cancer

Abstract The aim of this study was to construct two plasmid-specific shRNA transcripts of the bcl-2 gene in order to prepare for reverse of cell apoptosis. The plasmid was designed according to a previously published sequence of interfering RNA following an appropriate reference, using appropriate software. By annulling complementary oligonucleotides, double-stranded inserts were formed. Recombinant shRNA-encoding plasmids were constructed by digestion of psiRNA-x7SKGFPzeo plasmid (psiRNA-x7SKGFPzeo, with restrictive endonuclease BbsI electrophoresis in ultra-pure agarose with low melting point (LMP-Agarose). For each of the constructs, a suitable double-stranded insert downstream of x7SK (strong RNA III promoter) with T4 DNA ligase was cloned. The control plasmid psiRNAScr was used directly for transformation. The PC-3 cell lines were transfected with 2 plasmids, psiRNA-Bcl-2 and psiRNAScr to suppress the bcl-2 gene construct. The results have shown that the lowest level of bcl-2 genes was 48 h, and even lower 72 h after the transfer, and the mRNA levels returned to normal in 120 h. An increase in the percentage of cells with spontaneous apoptosis has been observed with successful inhibition of the bcl-2 gene. The induction of apoptosis in transfected cells increased the percentage of necrotic cells proportionally. The percentage of apoptotic cells transfected with psiRNA-bcl-2 plasmid increased proportionally to the increase of hydrogen peroxide concentration. The transfection of the PC-3 cell line from prostate cancer with constructed shRNA plasmid has induced suppression of bcl-2 gene expression versus control Scr plasmid. Suppression of bcl-2 gene expression significantly increased cell sensitivity to apoptosis induction.

Effect of the most common wound antiseptics on human skin fibroblasts.

BackgroundAntiseptics are used for the cleansing of acute or chronic wounds to eliminate micro‐organisms from the wound bed. However, they have effects on the skin cells.AimTo determine the effects of hexetidine, povidone–iodine (PI), undecylenamidopropyl‐betaine/polyhexanide (UBP), chlorhexidine, disodium eosin and hydrogen peroxide on human skin fibroblasts.MethodsCCD‐1064Sk cells were treated with hexetidine, PI, UBP, chlorhexidine, disodium eosin or hydrogen peroxide. Spectrophotometry was used to measure cell viability and flow cytometry was used to study apoptosis and necrosis after the treatment. In vitro wound scratch assays were performed to determine the gap closure.ResultsAll antiseptics significantly reduced the viability of human skin fibroblasts compared with controls. The percentage wound closure was lower with hexetidine, PI and UBP. The scratch assay could not be measured after treatments with chlorhexidine, disodium eosin or hydrogen peroxide, owing to their cytotoxicity. The apoptosis/necrosis experiments evidenced a significant reduction in viable cells compared with controls. An increased percentage of apoptotic cells was observed after treatment with all antiseptics. Compared with controls, the percentage of necrotic cells was significantly increased with all antiseptics except for hexetidine.ConclusionThe proliferation, migration and viability of human skin fibroblasts are reduced by treatment with hexetidine, PI, UBP, chlorhexidine, disodium eosin and hydrogen peroxide.

Puerarin mitigates oxidative injuries, opening of mitochondrial permeability transition pores and pathological damage associated with liver and kidney in Xanthium strumarium-intoxicated rats

In this study, the therapeutic effects of puerarin on Xanthium strumarium toxicity, which can develop in many species and does not have a specific antidote, were investigated. A single dose of 100 g/kg X. strumarium seeds was administered by gavage to female Sprague-Dawley rats, 6 h following which 200 mg/kg puerarin was administered by the same route, with puerarin administration being repeated daily at the same time. After completing the application, the blood, liver and kidney tissues of the rats were examined. Further, the biochemical parameters, glucose, MDA, GSH, SOD, mitochondrial Ca2+ and mitochondrial permeability transition pore (mPTP) opening levels, apoptotic factors (TUNEL, Bax and Bcl-2), ATP synthase and histopathological changes of the experimental rats were examined. The results revealed that while the administration of X. strumarium resulted in increased blood AST, ALT, ALP, LDH, CK, BUN and creatinine levels, it decreased glucose levels. In addition, it increased the MDA levels in the tissues and significantly increased the oxidative stress levels by decreasing the GSH levels and SOD activity. X. strumarium caused an increase in the mitochondrial Ca2+ and mPTP opening levels. Moreover, it increased the immunohistochemically determined ATP synthase expression and histopathologically identified necrotic liver cell death rates. Owing to its antioxidant properties and inhibitory effects on mPTP opening, puerarin administered for therapeutic purposes decreased the oxidative damage caused by X. strumarium toxicity, blood biochemical parameter levels, mitochondrial Ca2+ levels, mPTP opening, ATP synthase expression and the percentage of necrotic cells. Hence, the reduction in the liver and kidney damage in X. strumarium toxicity by puerarin indicates its potential use as an antidote for X. strumarium poisoning.

Preparation and characterization of amine-functionalized mupirocin-loaded zinc oxide nanoparticles: A potent drug delivery agent in targeting human epidermoid carcinoma (A431) cells

Skin cancer is one of the conventional malignities hastened in humans associated with sunburns, exhibiting considerable morbidity rate disparity. Besides the current treatments, nanotechnology alongside combination therapy holds enormous potential in treating metastatic tumors by selectively targeting cell surface receptors with a suitable carrier for drug delivery. The primary purpose of this research involves investigating and scrutinizing the physicochemical characteristics and antiproliferative effects of amine-functionalized and mupirocin-loaded zinc oxide nanoparticles (AZNP-MUP). Physicochemical characterization of the AZNP-MUP revealed spherical NPs (18.6 nm) on the drug-loaded matrix developed by EDC/NHS mediation, with an average crystallite grain size of 24.75 nm. The 50% hydroxyl radical scavenging activity for the conjugate (AZNP-MUP) was achieved at 91.89 μg/ml, the best concentration evinced when juxtaposed with the other antioxidant assays in the study. Moreover, AZNP-MUP inhibited the growth of V. cholera, E. faecalis and L. monocytogenes more significantly, with a greater zone of inhibition (25 mm) than AZNP. The conjugate AZNP-MUP manifested a strong antiproliferative effect with a higher percentage of necrotic cells (63%) and more DNA fragmentation than AZNP. Our results evince that AZNP-MUP could be adeptly proposed as a skin cancer therapy strategy to improve current patients' treatments.

Insights into the activation of oral keratinocyte cell death by Candida albicans and Staphylococcus aureus biofilms

Polymicrobial biofilms comprising Candida albicans and Staphylococcus aureus can increase the frequency and severity of oral diseases. This study assessed oral keratinocyte cell death, apoptosis and/or necrosis, promoted by soluble factors from single and dual biofilms of S. aureus and C. albicans. The soluble factors were obtained from the 16-h biofilm growth media. Cell viability was assessed by MTT and cell membrane damage by LDH. SEM was used for morphology changes. Assessment of apoptosis and necrosis was performed using annexin V and propidium iodide and caspases -2, -3, -6, -8 and -9. Statistical analysis was conducted with ANOVA and Tukey tests (α = 5%). Dual biofilms promoted the greatest harmful effect on oral cells, with a viability rate of 31.76%, damage to cell membranes and LDH released. Dual biofilms also induced higher percentages of necrotic cells (24.95%). Apoptosis was associated with caspases -2, -3, -6 and -8 activation.

Evaluation of Silver Nanoparticles Caffeic Acid Complex Compound as New Potential Therapeutic Agent against Cancer Incidence in Mice.

Objective:The present work was designed to study the effect of new conjugated caffeic and folic acid with silver nanoparticles with definite molecular size applied with and without gamma radiation exposure, as an antitumor agent against experimentally induced Ehrlich tumor and attempted to identify their potential molecular mechanisms of action throughout determination of anti-tumor activities using MTT cytotoxic assay against two human carcinoma cell lines in vitro, such as apoptosis analysis by flow cytometry through caspase-8, caspase-3 and TNF determination in vivo. Materials and Methods:Adult female albino mice were used and divided into five groups. Animals were sacrificed and the following parameters were estimated, glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD) in blood in addition to caspase8, caspase 3 and tumor necrosis factor (TNF) of tumor tissue, liver and kidney function also measured in plasma. The tumor specimens were processed for histopathological examination. Results:Nano-silver folate caffeic (NSFC) complex compound treatment resulted in growth inhibition in Ehrlich solid tumor, Hep-G2, and MCF-7 cells (IC50 0.062 mg, 7.70 µM, and 14.50 µM, respectively). Flow cytometric analysis revealed that (NSFC) with radiation IR had apoptotic effects at caspases 8 (Mean±SD) (49.4±14), caspase3 (39.97±9.75), and TNF (40.1±3.4) more than any other groups. Those disturbances were found to be associated with a kinetic induction of apoptosis and showed modulation of the antioxidant system {glutathione (GSH), glutathione peroxidase (GPx) and superoxide dismutase (SOD) which were 60.70±0.80, 26.73±0.80, 39.52±0.58 respectively}at the group which took (NSFC+IR), besides its high percentage of necrotic cells by histopathological studies. In conclusion,the present study showed that the treatment of (NSFC) exhibits very efficient oncolytic activity in delaying tumor growth in mice bearing Ehrlich Solid Carcinoma (ESC) and the mechanisms underlying the inhibitory effect of the present compound involve both an apoptotic effect against Hep-G2 and MCF-7 cells and modulation of antioxidant system.

Assessment of Anti-Cancerous Effect of Green, Roasted and Decaffeinated Coffee on Oral Squamous Cell Carcinoma Cell Line (In Vitro Study)

Oral Squamous Cell Carcinoma (OSCC) is one of the most prevailing malignancies of the head and neck area. So far, treatment methods are associated with harmful effects which drive attention towards natural compounds such as coffee. The current study attempted to test the anti-cancerous effect of green coffee, medium roasted coffee and decaffeinated coffee in OSCC cell lines and correlate the obtained results with their total phenolic content. We prepared coffee extracts using soxhlet apparatus. Then, we purchased and sub-cultured OSCC-25 cell into four study groups. We subjected three of those groups to coffee extracts separately. We left the remaining group without any treatment as a control group. We assessed cell cytotoxic effect of each extract by MTT viability assay. Additionally, we evaluated the apoptotic effect of each extract on OSCC-25 cell lines using flowcytometric analysis. Finally, we measured the total phenolic content in each coffee extract. The results revealed that coffee extracts induced varying degrees of cell cytotoxicity and apoptosis. Green coffee showed the highest cytotoxic and apoptotic effect followed by medium roasted coffee, then decaffeinated coffee. The total percentage of necrotic cells were higher in the coffee extracts groups, compared to the control group with a higher value in favor of green coffee followed by medium roasted coffee. Upon chemical analysis, the results showed that green coffee extract contained the highest concentration of phenolic compounds followed by medium roasted coffee then decaffeinated coffee extracts. We concluded that green coffee was the most potent anti-cancer extract. It seems plausible that coffee, and particularly green coffee could be for treating of OSCC.

Selected Kefir Water from Malaysia Attenuates Hydrogen Peroxide-Induced Oxidative Stress by Upregulating Endogenous Antioxidant Levels in SH-SY5Y Neuroblastoma Cells.

Kefir, a fermented probiotic drink was tested for its potential anti-oxidative, anti-apoptotic, and neuroprotective effects to attenuate cellular oxidative stress on human SH-SY5Y neuroblastoma cells. Here, the antioxidant potentials of the six different kefir water samples were analysed by total phenolic content (TPC), total flavonoid content (TFC), ferric reducing antioxidant power (FRAP), and 2,2′-diphenyl-1-picrylhydrazyl radical (DPPH) assays, whereas the anti-apoptotic activity on hydrogen peroxide (H2O2) induced SH-SY5Y cells was examined using MTT, AO/PI double staining, and PI/Annexin V-FITC assays. The surface and internal morphological features of SH-SY5Y cells were studied using scanning and transmission electron microscopy. The results indicate that Kefir B showed the higher TPC (1.96 ± 0.54 µg GAE/µL), TFC (1.09 ± 0.02 µg CAT eq/µL), FRAP (19.68 ± 0.11 mM FRAP eq/50 µL), and DPPH (0.45 ± 0.06 mg/mL) activities compared to the other kefir samples. The MTT and PI/Annexin V-FITC assays showed that Kefir B pre-treatment at 10 mg/mL for 48 h resulted in greater cytoprotection (97.04%), and a significantly lower percentage of necrotic cells (7.79%), respectively. The Kefir B pre-treatment also resulted in greater protection to cytoplasmic and cytoskeleton inclusion, along with the conservation of the surface morphological features and the overall integrity of SH-SY5Y cells. Our findings indicate that the anti-oxidative, anti-apoptosis, and neuroprotective effects of kefir were mediated via the upregulation of SOD and catalase, as well as the modulation of apoptotic genes (Tp73, Bax, and Bcl-2).

Evaluation of Silver Nanoparticles Caffeic folate (NSFC) Complex Compound as New Potential Therapeutic Agent against Cancer Xenograft in Mice: Thesis Abstract

Thesis Abstract In the last few decades the properties of silver compounds have been of interest as potential cancer treatments. The present work was designed to study the effect of new conjugated caffeic and folic acid with silver nanoparticle with definite molecular size applied with and without gamma radiation exposure, as an antitumor agent against experimentally induced Ehrlich tumor and attempted to identify their potential molecular mechanisms of action throughout determination of anti-tumor activities using MTT cytotoxic assay against two human carcinoma cell lines in vitro, Apoptosis analysis by flow cytometry through caspase-8, caspase-3 and TNF determine in vivo . Adult female albino mice were used and divided into five groups, Animals were scarified and the following parameters were estimated, glutathione, glutathione peroxidase, superoxide dismutase in blood and caspase8, caspase 3 and TNF from scarified tumor tissue. The tumor specimens were processed for histopathological examination. Nano-silver folate caffeic (NSFC) complex compound treatment resulted in a growth inhibition in Hep-G2 and MCF-7 cells (IC50 7.70 µM and 14.50 µM, respectively). Flow cytometric analysis revealed that (NSFC) with radiation exposed has apoptotic effect at caspases 8, 3 and TNF more than any compound of them alone. That disturbance was found to be associated with a kinetic induction of apoptosis and showed modulation of antioxidant system beside its high percentage of necrotic cells by histopathological studies. the novel synthetic Nano-silver folate Caffeic complex compound may potentially present a new hope for the development of breast cancer therapeutics, which should attract further scientific and pharmaceutical interest.

The Encapsulation of Febuxostat into Emulsomes Strongly Enhances the Cytotoxic Potential of the Drug on HCT 116 Colon Cancer Cells.

Febuxostat (FBX) is a drug able to inhibit xanthine oxidase and reduce uric acid production commonly used for the treatment of hyperuricemia in subjects suffering from gout. Several studies have also been directed at its use as anti-cancer drug during the last years, opening a window for its off-label use. In the present study, an optimized formulation in terms of vesicle size and drug release, obtained by encapsulation of FBX into the emulsomes (FBX-EMLs), was evaluated for its cytotoxic potential in human colorectal carcinoma (HCT 116) cells. The optimized FBX-EMLs formula had an improved half maximal inhibitory concentration (IC50), about 4-fold lower, compared to the free drug. The cell cycle analysis showed a significant inhibition of the HCT 116 cells proliferation following FBX-EMLs treatment compared to all the other conditions, with a higher number of cells accumulating on G2/M and pre-G1 phases, paralleled by a significant reduction of cells in G0/G1 and S phases. The optimized formula was also able to significantly increase the percentage of cell population in both early and late stages of apoptosis, characterized by a higher intracellular caspase-3 concentration, as well as percentage of necrotic cells. Lastly, the FBX ability to decrease the mitochondrial membrane potential was enhanced when the drug was delivered into the EMLs. In conclusion, the new formulation of FBX into EMLs improved all the parameters related to the anti-proliferative activity and the toxic potential of the drug towards colorectal cancer cells.