Apoptotic Changes Research Articles

Role of Fasting Mimicking Diet in Farnesoid x Receptor for Suppressing Epithelial-to-Mesenchymal Transition, Cell Cycle Progression, and Viability of Prostate Cancer Cells

The systemic and resistant nature of metastatic castration-resistant prostate cancers (mCRPC) renders it largely incurable even after intensive multimodal therapy. Proliferation, survival, and epithelial-mesenchymal transition (EMT) are three fundamental events that are deeply linked to carcinogenesis. Hence, it is necessary to find a new combination of several therapies, targeting those vital mechanisms without causing side effects. Significant research works have shown differential low expression of the metabolic Farnesoid X receptor (FXR) in primary and metastatic prostate cancer suggesting their importance in prostate pathogenesis. Obticholic acid (INT 747), a potent FXR agonist is widely used in primary biliary cholangitis, and Fasting mimicking Diet (FMD) both were drastically showed effects on different cancer progression. We hypothesized that FXR and FMD may inhibit proliferation and the metastatic phenotype in PC-3 prostate cancer cells. Analyses of the cell viability, cell cycle, migration, and matrigel invasion assays were performed to elucidate how INT 747 and /or FMD functions in prostate cancer. In this study, INT 747 treatment caused apoptotic morphological changes and significantly reduced the survival of PC-3 cells incubated in normal mediums. Furthermore, we showed that the combination of the INT 747 and FMD was much more harmful to cancer cells than the treatment with INT 747 or FMD alone. Moreover, our study showed that INT 747 either alone or combined with FMD robustly induced cell cycle arrest at the S phase. Interestingly, the combination treatment on PC-3 cells not only showed several lines of evidence of apoptotic cells death but also inhibited carcinogenic potential as evaluated by impairment of spheroid formation capacity and delayed wound healing and matrigel invasion. At the cellular level, FXR activation resulted in down-regulation of procaspase -3, vimentin, and MMP9, which triggers apoptotic cell death, cell cycle arrest, and switch from mesenchymal to an epithelial phenotype. Collectively, FXR activation alone markedly decreases, and when combined with FMD abrogates the survival and carcinogenic potential of metastatic prostate cancer cells.

KLK4 affects the biological behavior of oral squamous cell carcinoma through Wnt/β catenin and PI3K/Akt signaling pathways

Purpose: To investigate the effect of kallikrein 4 (KLK4) on oral squamous cell carcinoma (OSCC), and its mechanism of action.
 Method: Human tongue SCC cell line TCA-8113 was cultured, and the interference fragment and irrelevant sequence fragment of KLK4 gene were designed and synthesized. These fragments were constructed into pgcsi-h1 plasmid. Then, the TCA-8113 cells were cultured for 24 h, followed by transfection, after which TCA-8113-NC and TCA-8113-siKLK4 cells were obtained. The mRNA expression of KLK4 was determined using quantitative real-time-polymerase chain reaction (PCR). Proliferative potential was assessed using MTT method, while apoptosis was determined by flow cytometry. Furthermore, cell migration was evaluated by scratch test, and Transwell cell invasion experiment was used to determine cell invasion. Immunoblot assay was applied to determine the expressions of E- and N-cadherins, vimentin, uPA, Wnt1, β-Catenin, p-pi3k and p-Akt.
 Result: Compared with TCA-8113-nc, the expression of KLK4 in TCA-8113-siKLK4 cells was significantly down-regulated, while, relative to TCA-8113-NC, TCA-8113-siKLK4 cells had significantly reduced cell proliferation, invasion and migration, and enhanced apoptosis (p < 0.05). Moreover, there was significant up-regulation of E- and N-cadherins, vimentin, uPA, Wnt1, β-Catenin, p-pi3k and p-Akt in TCA-8113-siKLK4 group, relative to TCA-8113-NC (p < 0.05).
 Conclusion: KLK4 enhances the multiplication, metastasis and invasive potential of OSCC cells, but suppresses apoptotic changes via activation of Wnt/β-catenin/PI3K/Akt signal route. The potential of Kallikrein-4 as biomarker for OSCC cells should be determined in suitable animal models.

Modulatory role of Coriandrum sativum (coriander) extract against diabetic complications on the gonads of female rats and their offspring

It is well known that diabetes is associated with impairment of ovarian and testicular structure and function. Coriander (Coriandrum sativum L.) is identified as one of the oldest herbal plants valued for its nutritional and medicinal properties. This work is mainly designed to evaluate the possible modulatory role of dry coriander fruit extract against gonadal impairments associated with diabetes in female rats and their pups. Twenty-four pregnant rats were divided into four groups (n = 6): group I served as control, group II was administered daily with coriander fruit extract (250 mg/kg b.wt), group III was injected interaperitoneally with a single dose of streptozotocin (STZ) (80 mg/kg b.wt), and group IV was injected with single dose of STZ and post administered coriander extract. The experiment was conducted from the 4th day of gestation till the end of weaning. At the end of the experiment, the mothers’ rats and their offspring were weighed, sacrificed, the ovaries from mothers as well as ovaries and testes from offspring were immediately excised, and processed for histological, immunohistochemical and evaluation of apoptosis and transforming growth factorβ (TGF-β). Also, blood samples were collected and analyzed to estimate the levels of sex hormones as well as antioxidants.In STZ induced diabetes in mother's rats and their offspring, the ovarian sections revealed severe histopathological signs included several atretic follicles, dilated and congested blood capillaries. Additionally, the testicular sections of offspring appeared with destructive seminiferous tubules. Immunohistochemically, the ovarian sections displayed weak to negative expression for calretinin marker however the testicular sections showed strong expression for Bax protein (apoptotic marker) and weak to negative expression for Ki67 protein (proliferative marker). Also, the mean % values of positively expressed cells for TGF-β and annexin-v markers (late and early apoptosis indicator) were significantly elevated in the ovarian and testicular tissues of STZ-induced group of mother's rats and their pups if compared with control. Further results revealed that the levels of insulin, FSH, LH, estrogen, SOD and CAT were significantly decreased if compared with control however the levels of MDA and NO were significantly elevated. Administration of coriander fruit extract to diabetic rats successfully alleviated most of the altered histological, immunohistochemical, biochemical, and apoptotic changes induced by diabetes. Coriandrum sativum fruit extract has a powerful ameliorative role against STZ-induced diabetic gonadal dysfunctions in female rats and their offspring.

Plantamajoside alleviates acute sepsis-induced organ dysfunction through inhibiting the TRAF6/NF-κB axis

Context Plantamajoside (PMS) possesses rich pharmacological characteristics that have been applied to remedy dozens of diseases. However, the understanding of PMS in sepsis remains insufficient. Objective Role of PMS in sepsis-regulated organ dysfunction and potential mechanisms were investigated. Materials and methods Thirty C57BL/6 male mice were adaptive fed for three days and used to establish acute sepsis model by caecal ligation and perforation (CLP). These experimental mice were divided into Sham, CLP, CLP + 25 mg PMS/kg body weight (PMS/kg), CLP + 50 mg PMS/kg and CLP + 100 mg PMS/kg (n = 6). The pathological and apoptotic changes of lung, liver and heart tissues were observed via HE and TUNEL staining. The injury-related factors of lung, liver and heart were detected by corresponding kits. ELISA and qRT-PCR were applied to assess IL-6/TNF-α/IL-1β levels. Apoptosis-related and TRAF6/NF-κB-related proteins were determined using Western blotting. Results All doses of PMS enhanced the survival rates in the sepsis-induced mouse model. PMS remitted sepsis-mediated lung, liver and heart injury through prohibiting MPO/BALF (70.4%/85.6%), AST/ALT (74.7%/62.7%) and CK-MB/CK (62.3%/68.9%) levels. Moreover, the apoptosis index (lung 61.9%, liver 50.2%, heart 55.7% reduction) and IL-6/TNF-α/IL-1β levels were suppressed by PMS. Furthermore, PMS lowered TRAF6 and p-NF-κB p65 levels, whereas TRAF6 overexpression reversed the protective influences of PMS in organ injury, apoptosis and inflammation triggered by sepsis. Discussion and conclusions PMS suppressed sepsis-induced organ dysfunction by regulating the TRAF6/NF-κB axis, and PMS treatment may be considered as a novel strategy for sepsis-caused damage in future.

Elemene inhibits the growth and promotes apoptosis of bladder cancer cells through PTEN-Akt signaling pathway

Purpose: To investigate the influence of elemene on growth and apoptotic changes in bladder cancer cells, and the involvement of the phosphatase and tensin homologous protein (PTEN)-serine/threonine kinase (Akt) signal pathway in the process.
 Methods: Human bladder cell line SV-HUC-1 and human bladder cancer cell line T24 were randomly assigned to blank control group and elemene groups. Protein expression levels of apoptosis-related factors (Bcl-2, Bax, and caspase-3] and PTEN, Akt, and p-Akt in both cells were determined using western blot assay.
 Results: In T24 cells, the protein level of Bcl-2 was significantly higher in elemene group than in blank control, while the protein expression levels of Bax and caspase-3 in elemene group were significantly lower than the corresponding levels in blank control group (p < 0.05). The protein expression of PTEN level in T24 cells was significantly higher in elemene group than in blank control group, while protein expression level of p-Akt was significantly lower in elemene group than in blank control (p < 0.05). The Akt protein expression was comparable in elemene and blank control groups.
 Conclusion: Elemene inhibits the growth and enhances apoptosis of bladder cancer cells through a mechanism involving up-regulation of the expression of PTEN and suppression of the expression of pAkt.

The effects of non-functionalized polystyrene nanoparticles of different diameters on the induction of apoptosis and mTOR level in human peripheral blood mononuclear cells

Particles of various types of plastics, including polystyrene nanoparticles (PS-NPs), have been determined in human blood, placenta, and lungs. These findings suggest a potential detrimental effect of PS-NPs on bloodstream cells. The purpose of this study was to assess the mechanism underlying PS-NPs-induced apoptosis in human peripheral blood mononuclear cells (PBMCs). Non-functionalized PS-NPs of three diameters: 29 nm, 44 nm, and 72 nm were studied used in this research. PBMCs were isolated from human leukocyte–platelet buffy coat and treated with PS-NPs at concentrations ranging from 0.001 to 200 μg/mL for 24 h. Apoptotic mechanism of action was evaluated by determining the level of cytosolic calcium ions, as well as mitochondrial transmembrane potential, and ATP levels. Furthermore, detection of caspase-8, -9, and -3 activation, as well as mTOR level was conducted. The presence of apoptotic PBMCs was confirmed by the method of double staining of the cells with propidium iodide and FITC-conjugated Annexin V. We found that all tested NPs increased calcium ion and depleted mitochondrial transmembrane potential levels. The tested NPs also activated caspase-9 and caspase-3, and the smallest NPs of 29 nm of diameter also activated caspase-8. The results clearly showed that apoptotic changes and an increase of mTOR level depended on the size of the tested NPs, while the smallest particles caused the greatest alterations. PS-NPs of 26 nm of diameter activated the extrinsic pathway (increased caspase-8 activity), as well as intrinsic (mitochondrial) pathway (increased caspase-9 activity, raised calcium ion level, and decreased transmembrane mitochondrial potential) of apoptosis. All PS-NPs increased mTOR level at the concentrations smaller than those that induced apoptosis and its level returned to control value when the process of apoptosis escalated.

Restorative effects of gallic acid against sub-chronic hepatic toxicity of co-exposure to zinc oxide nanoparticles and arsenic trioxide in male rats

Background and objectivesThis study aimed to assess the effect of zinc oxide nanoparticles (ZNPs) and/or arsenic trioxide (ATO) exposure on the liver of adult male Sprague Dawley rats. Moreover, the probable ameliorative impact of gallic acid (GA) against ZNPs and ATO-induced hepatotoxicity and the possible underlying mechanisms were evaluated. MethodsSixty male Sprague Dawley rats were distributed into six groups. The 1st and 2nd groups were orally given distilled water (1 ml/kg) and 20 mg GA/kg b. wt, respectively. The 3rd and 4th groups were orally given 100 mg ZNPs/kg b. wt and 8 mg ATO/kg b. wt, respectively. The 5th group was co-administered ZNPs and ATO at the doses mentioned above. The last one was co-administered ZNPs, ATO, and GA at the earlier described doses. All tested compounds were orally given once a day for 60 successive days. Then, serum levels of alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total, direct, indirect bilirubin, triglycerides, total cholesterol, HDL, VLDL, and LDL were estimated. The hepatic content of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) was evaluated. Moreover, Bcl-2 and Bax's reactive proteins were immunohistochemically detected, and Zn and As residual patterns in hepatic tissues were assessed. ResultsZNPs, ATO, and ZNPs+ATO-exposed rats showed significantly (P < 0.001) elevated serum AST (219%, 233%, and 333%), ALT (300%, 400%, and 475%), ALP (169%, 205%, and 294%), and total bilirubin (42%, 68%, and 109%) compared to the control ones. On the other hand, a significantly (P < 0.001) declined SOD (58%, 49%, and 43%) and GPx (70%, 63%, and 56%) but increased MDA (133%, 150%, and 224%) was recorded in the hepatic tissues of ZNPs, ATO, and ZNPs+ATO exposed rats, respectively, relative to the control rats. Moreover, the hepatic tissues of the ZNPs, ATO, and ZNPs+ATO exposed rats showed a significant (P < 0.001) decrease in Bcl-2 (28%, 33%, and 23%) but elevation in Bax (217%, 267%, and 236%) immunoreactivities compared to the control rats. These findings were consistent with the microscopic alterations in the hepatic architecture and accumulation of Zn and As. Furthermore, a notable hyperlipidemic condition was recorded following ZNPs and/or ATO exposure. On the contrary, GA notably reduced hepatic enzymes compared to ZNPs+ATO-exposed rats. Additionally, GA markedly improved ZNPs+ATO-afforded liver tissue damage and apoptotic events. ConclusionOverall, GA oral dosing significantly mitigated the negative effects of ZNPs and ATO on the liver by improving the antioxidant defense system and controlling apoptotic changes.

Recent progress in apoptosis triggering facilitated by HeLa Studies

Abstract Objective: Cancer is a leading cause of death globally, prompting numerous efforts to find effective treatments. HeLa cells, derived from Henrietta Lacks’ cancerous squamous cells, have played a crucial role in cancer research due to their origin, resistance, and rapid growth. They are particularly useful for studying ways of cellular death triggering, or apoptosis, without an immune response. Thus, the objective of this paper was to review the latest publications on the subject of HeLa apoptosis so that a brief view to be available on the otherwise so extended subject. Methodology: To provide a concise review of the extensive research on this topic, a search was conducted using the phrase “HeLa cells apoptosis triggering” on PubMed. The articles that were published in English, in the last 6 years, presenting results sustained by valid morphological and chemical apoptotic changes present in cells, were selected and reviewed. A comprehensive table presenting the apoptotic mechanism exerted by each substance was made to assure a concise presentation of the results. Results: The reviewed studies have shown that many natural substances exhibit pro-apoptotic activity on malignant cells and can be used as chemotherapeutic agents. Some synthetic molecules were showed to have good results too. Important facts about these substances, their intervention site and metabolic modifications are presented in a concise form. The use of nano-carriers for targeted delivery was shown to increase their specificity towards cancerous cells. Conclusions: HeLa cells were a groundbreaking discovery that revolutionized scientific research. Although there is ongoing research towards cancer cures using HeLa cells, there are still many trials and considerations that need to be addressed. With the countless existing HeLa cell lines, the scientific possibilities for research are endless.

Apoptotic Changes, Oxidative Stress and Immunomodulatory Effects in the Liver of Japanese Seabass (Lateolabrax japonicus) Induced by Ammonia-Nitrogen Stress during Keep-Live Transport.

This study investigated the effects of NH3-N on antioxidant responses, histoarchitecture, and immunity of Japanese seabass (Lateolabrax japonicus) during keep-live transport. The findings suggest that NH3-N stress transport alters the transcription of P53, Caspase 9, Bcl2, Caspase 3 and Bax genes, demonstrating that NH3-N stress can trigger the apoptotic pathway of P53-Bax-Bcl2 and Caspase and induce apoptosis. NH3-N stress transport also evoked transcriptional upregulation of inflammatory cytokines (tumor necrosis factor α (TNF-α), Toll-like receptor 3 (TLR-3), nuclear factor kappa β (NF-κB), interleukin 6 (IL-6) and interleukin 1β (IL-1β)) and increased complement C3, C4, lysozyme (LZM) and immunoglobulin (IgM) levels, activating the innate immunological system during keep-live transport. In addition, NH3-N stress transport altered changes in the levels of superoxide dismutase (SOD), catalase (CAT), glutathione-related enzymes, and heat shock proteins 70 and 90 in the liver, indicating that the antioxidant system and Hsp protected the cells from NH3-N-induced oxidative stress. When excess ROS were not removed, they caused the body to respond with immunological and inflammatory responses, as well as apoptosis and tissue damage. This helps towards understanding the effect of NH3-N levels on sea bass during keep-live transport.

Male obesity is associated with sperm telomere shortening and aberrant mRNA expression of autophagy-related genes

BackgroundObesity is regarded a global public health crisis. It has been implicated in a variety of health problems, but when it comes to male fertility, how and to what extent obesity affects it are poorly understood.Accordingly, semen samples from 32 individuals with obesity (body mass index (BMI) ≥ 30 kg/m2) and 32 individuals with normal weight (BMI: 18.5-25 kg/m2) were obtained. Here, for the first time, we examined the association between obesity, relative sperm telomere length (STL) and autophagy-related mRNA levels such as Beclin1, AMPKa1, ULK1, BAX, and BCL2. Each group was also evaluated for conventional semen parameters, sperm apoptotic changes, DNA fragmentation index (DFI), sperm chromatin maturation, and reactive oxygen species (ROS) levels.ResultsBased on our findings, there was a marked reduction in relative STL in individuals with obesity as compared to the normal-weight group. We also found a significant negative correlation between relative STL and age, BMI, DFI, percentage of sperm with immature chromatin, and intracellular ROS levels in patients with obesity. In the normal-weight group, relative STL was only negatively correlated with DFI and intracellular ROS levels. Regarding mRNA expression, there was considerable upregulation of Beclin1, ULK1, and BCL2 in the group with obesity compared to the normal-weight group. Obesity was also found to be associated with a considerable decline in semen volume, total sperm count, progressive motility, and viability in comparison to normal-weight individuals. Furthermore, obesity was associated with considerably higher percentages of DFI, sperm with immature chromatin, late-stage apoptosis, and elevated ROS levels.ConclusionAccording to our findings, obesity is associated with sperm telomere shortening and aberrant autophagy-related mRNA expression. It should be emphasized that telomere shortening in sperm may be an indirect consequence of obesity due to the oxidative stress associated with the condition. Nevertheless, further investigation is required for a more comprehensive understanding.

Cytotoxic activity of essential oil from Leaves of Myrcia splendens against A549 Lung Cancer cells

BackgroundPlants of the Myrcia genus have been widely used in folk medicine to treat various diseases, including cancer. Myrcia splendens species has a diverse chemical constitution, but the biological activities of its essential oil have not been well investigated. In this study to out the chemistry characterization of essential oil (EO) from the leaves of the species M. splendens from Brazil and evaluate cytotoxic effect in A549 lung cancer cells.MethodsM. splendens EO was obtained by hydrodistillation and analyzed by Gas Chromatography-Mass Spectrometry (GC-MS). EO was isolated and evaluated for cellular viability in tumor cell lines by MTT assay. The evaluation of the formation of clones and the migratory capacity of the A549 cells treated with EO was done by the clonogenic assay and the wound healing assay. Morphological changes were observed in A549 cells by fluorescence using Phalloidin/FITC and DAPI.Results22 compounds were identified in the chemical analysis of EO, corresponding to 88% of the sample. Major compounds were the sesquiterpenic hydrocarbons bicyclogermacrene (15.4%), germacrene D (8.9%) and E-caryophyllene (10.1%). The biological analysis of the EO showed high cytotoxic activity with an IC50 below 20 µg/ml in the THP-1, A549 and B16-F10 tumor cells. The treatment with EO reduced colony formation and inhibited the migratory capacity of A549 cells. Furthermore, apoptotic morphological changes in the nucleus and cytoplasm of A549 cells was observed after of treatment with EO.ConclusionThe findings of this study suggest that the M. splendens EO has cytotoxic compounds for the A549 lung cancer cells. Treatment with the EO decreased the colony formation and reduced the ability of lung cancer cells to migrate. Future studies may be used to isolate compounds from the EO for the study of lung cancer.

In Vitro Assessment of the Impact of Ultraviolet B Radiation on Oral Healthy and Tumor Cells

Ultraviolet radiation (UVR) is generally considered a primary tumorigenic agent. While UVR exposure has been studied, especially at the skin level, the impact of UV exposure on internal tissues and its effect on the appearance and the development of tumors has not yet been fully examined. Although there are maximum limits for UVR exposure on external tissues, other internal tissues, such as oral tissue, can be exposed to UVR as well. Over the course of diagnosis and treatment, oral cells may be exposed to ultraviolet radiation; however, there has not been an established limit for UV radiation exposure. Therefore, the aim of the current study was to examine the effects of ultraviolet-B (UVB) radiation at two doses (2.5 and 5 J/cm2) on tumor cells (pharyngeal carcinoma and tongue carcinoma) and healthy cells (gingival fibroblasts). The viability of the cells and their morphology, actin filaments, and nuclei structures; the expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax) genes; and the roles of caspases-3/7, 8, and 9 were determined after the cells had been exposed to UVB. The experiments revealed that both types of cell lines showed reductions in viability, especially at a dose of 5 J/cm2. Additionally, apoptotic-like changes (rounding of the cells, the condensation of the nuclei, the re-organization of the actin filaments) were observed in all analyzed cells. The expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax) genes revealed that UVB (5 J/cm2) may induce apoptosis in both oral tumor and healthy cells. Moreover, an analysis of caspases-3/7, 8, and 9 showed that UVB exposure enhanced their activity, suggesting that cell death could be caused by both intrinsic and extrinsic apoptosis. Accordingly, UVB exposure at the maximum doses used in dental practices (5 J/cm2) induced nonselective apoptotic changes, thereby reducing both tumor and healthy cell viability.

Swimming training reduced inflammation and apoptotic changes in pulmonary tissue in type 1 diabetic mice.

Despite the vulnerability of pulmonary tissue to diabetic conditions, there are few reports related to the detrimental effects of hyperglycemia and therapeutic modalities on lung parenchyma. Here, the apoptotic changes were monitored in the diabetic pulmonary tissue of mice (DM1) subjected to a four‒week swimming plan. The mice were randomly allocated into Control; Control + Swimming (S); Diabetic group (D); and Diabetic + Swimming (D + S) groups (each in 8 mice). In the D and D + S groups, mice received intraperitoneally 50mg/kg of streptozotocin (STZ). After 14 days, swimming exercise was done for four weeks. The expression of il-1β, bcl-2, bax, and caspase-3 was investigated using real-time PCR analysis. A histological examination was performed using H&E staining. DM1 significantly upregulated il-1β, bax, and caspase-3, and down-regulated bcl-2 compared to the non-diabetic mice (p < 0.05). We noted that swimming exercises reversed the expression pattern of all genes in the diabetic mice and closed to basal levels (p < 0.05). Data indicated that swimming exercise could diminish emphysematous changes, and interstitial pneumonitis induced by STZ. Along with these changes, swimming exercise had protective effects to reduce the thickness of the inter-alveolar septum and mean alveolar area in diabetic mice. These data demonstrated that swimming exercises could decrease DM1-related pathologies in mouse lungs by regulating apoptosis and inflammatory response.

Glucose-conjugated glutenin nanoparticles for selective targeting and delivery of camptothecin into breast cancer cells.

Receptor-mediated drug delivery systems are a promising tool for targeting malignant cells to suppress/inhibit the malignancy without disturbing healthy cells. Protein-based nanocarrier systems possess numerous advantages for the delivery of variety of chemotherapeutics, including therapeutic peptides and genes. In the present work, glucose-conjugated camptothecin-loaded glutenin nanoparticles (Glu-CPT-glutenin NPs) were fabricated to deliver camptothecin to MCF-7 cells via GLUT-1 transporter protein. Initially, Glu-conjugated glutenin polymer was successfully synthesized through reductive amination reaction, and this was confirmed by FTIR and 13C-NMR. Then, camptothecin (CPT) was loaded into Glu-conjugated glutenin polymer forming Glu-CPT-glutenin NPs. The nanoparticles were studied for their drug releasing capacity, morphological shape, size, physical nature, and zeta potential. The fabricated Glu-CPT-glutenin NPs were found to be spherical in shape and amorphous in nature with 200-nm size range and a zeta potential of - 30mV. Furthermore, MTT assay using Glu-CPT-glutenin NPs confirmed concentration-dependent cytotoxicity against MCF-7 cells after 24-h treatment, and IC50 was found to be 18.23μgmL-1. In vitro cellular uptake study demonstrated that the Glu-CPT-glutenin NPs had enhanced endocytosis and delivered CPT in MCF-7 cells. A typical apoptotic morphological change of condensed nuclei and distorted membrane bodies was found after treatment with IC50 concentration of NPs. The released CPT from NPs also targeted mitochondria of MCF-7 cells, significantly increasing the level of reactive oxygen species and causing the damage of mitochondrial membrane integrity. These outcomes confirmed that the wheat glutenin can positively serve as a significant delivery vehicle and enhance the anticancer potential of this drug.

Co-administration of exercise training and melatonin on the function of diabetic heart tissue: a systematic review and meta-analysis of rodent models

PurposeDiabetes mellitus (DM), a hyperglycemic condition, occurs due to the failure of insulin secretion and resistance. This study investigated the combined effects of exercise training and melatonin (Mel) on the function of heart tissue in diabetic rodent models.MethodsA systematic search was conducted in Embase, ProQuest, Cochrane library, Clinicaltrial.gov, WHO, Google Scholar, PubMed, Ovid, Scopus, Web of Science, Ongoing Trials Registers, and Conference Proceedings in July 2022 with no limit of date or language. All trials associated with the effect of Mel and exercise in diabetic rodent models were included. Of the 962 relevant publications, 58 studies met our inclusion criteria as follows; Mel and type 1 DM (16 studies), Mel and type 2 DM (6 studies), exercise and type 1 DM (24 studies), and exercise and type 2 DM (12 studies). Meta-analysis of the data was done using the Mantel Haenszel method.ResultsIn most of these studies, antioxidant status and oxidative stress, inflammatory response, apoptosis rate, lipid profiles, and glucose levels were monitored in diabetic heart tissue. According to our findings, both Mel and exercise can improve antioxidant capacity by activating antioxidant enzymes compared to the control diabetic groups (p < 0.05). The levels of pro-inflammatory cytokines, especially TNF-α were reduced in diabetic rodents after being treated with Mel and exercise. Apoptotic changes were diminished in diabetic rodents subjected to the Mel regime and exercise in which p53 levels and the activity of Caspases reached near normal levels (p < 0.05). Based on the data, both Mel and exercise can change the lipid profile in diabetic rodents, especially rats, and close it to near-to-control levels.ConclusionThese data showed that exercise and Mel can reduce the harmful effects of diabetic conditions on the heart through the regulation of lipid profile, antioxidant capacity, apoptosis, and inflammation.

140 Diffuse Midline Glioma Harbors Alterations That are Attenuated by MEK Inhibitors and 5-Aminolevulinic Acid-Photodynamic Therapy

INTRODUCTION: Diffuse midline glioma (DMG) is the leading cause of brain cancer-related mortality in children. There is a desperate need for therapeutic models that abrogate tumor cell invasion processes since standard radiation therapy is associated with transitory benefit and therapeutic resistance. 5-aminolevulinic acid-photodynamic therapy (5-ALA-PDT) is a photo-therapy that preferentially ablates tumor cells when 5-ALA is delivered with 635 nm light by metabolizing to protoporphyrin IX (5-ALA metabolite). The RAS/MEK signaling pathway has been implicated in tumorigenesis and been shown to synergize with 5-ALA-PDT in various carcinoma models. METHODS: Five DMG lines were treated with 5-ALA-PDT and MEKi trametinib to evaluate effects on proliferation, invasion, live cell dynamics, and cell death in vitro. Apoptotic effects and changes in the tumor microenvironment were evaluated in vivo using a patient-derived xenograft model RESULTS: Functional screens demonstrate sensitivity to 5-ALA-PDT and trametinib (5-ALA-PDT IC50 = 0.25-0.41 µM; trametinib IC50 = 0.03 µM-0.03 mM). An increase in cleaved-caspase3 (p = 0.0006) and reactive oxygen species concentration (p &lt; 0.0001) was characteristic in DMG cell lines post-treatment. Immunofluorescence stainings showed a decrease in PDGFRa (p = 0.007) and increase in Hsp70 (p &lt; 0.0001) expression. Trametinib and 5-ALA-PDT dampened cell invasion (invasive/total cells: 1.30, 0.20, 0.26, 0.05 for control, trametinib, 5-ALA-PDT, and combination, respectively). For in vivo studies, there were changes in cleaved-caspase3 (&gt;4-fold difference) and glioma-associated neovascularization in tumor bearing mice. CONCLUSIONS: This study proposes a promising, novel combinatorial strategy involving the use of 5-ALA-PDT and MEKi for DMG treatment and can be implemented for other CNS tumors.

TITLE OF THE ARTICLE: EXOINTERACTIVE AMELIORATION AGAINST METHOTREXATE INTOXICATION IN MOUSE SPERMATOZOA

Objective: To assess the synergistic potential using an exointeractive association between Murraya koenigii hydromethanolic leaf extract and the synthetic compound quercetin to offset methotrexate induced toxicity in mouse cauda epididymal spermatozoa. Methods: Healthy adult male albino mice were divided into eight groups (N= 6/group) and were treated with methotrexate intraperitoneally (10mg/kg body weight) at intervals of every 48 hours for 15 days, to combat this toxic effects of the drug, Murraya koenigii hydromethanolic leaf extract (250mg/kg body weight) and quercetin (15mg and 30mg) alone as well as in combination, were given by oral administration. Various sperm parameters such as sperm count, motility and viability followed by Hypo-osmotic Swelling (HOS) test, apoptosis test, Sperm Toroid Integrity (STI) and Sperm Chromatin Dispersion (SCD) assay. Statistical analysis used: Statistical analysis was carried out using Graphpad Prism Version 7 to verify levels of significance at the P &lt; 0.001 value in one-way ANOVA. Results: Methotrexate induced a substantial reduction in sperm count, motility and viability which induces apoptotic changes, also resulted in disruption of sperm toroid protamine-DNA binding, increased fragmentation of nuclear DNA by decreased sperm chromatin dispersion and low percentage of swollen sperm representing membrane integrity loss. whereas Murraya koenigii hydromethanolic leaf extract and quercetin proved effective in reversing the drug induced toxicity, especially with the 15 mg dose of quercetin along with the plant extract brought about significant amelioration than 30mg dose of quercetin due to its pro-oxidant effects. Conclusions: To reduce the toxic manifestations of methotrexate, this exointeractive approach proved efficient.

Prospective Analysis of B Lymphocyte Subtypes, before and after Initiation of Dialysis, in Patients with End-Stage Renal Disease.

CD5, CD27, BAFF, IgM and annexin were evaluated by flow cytometry on CD19+ cells in ESRD patients (n = 40), at time of initiating HD or CAPD (T0) and 6 months later (T6). A significant reduction in ESRD-T0 compared to controls was noticed for CD19+, 70.8 (46.5) vs. 171 (249), p < 0.0001, CD19+CD5-, 68.6 (43) vs. 168.9 (106), p < 0.0001, CD19+CD27-, 31.2 (22.1) vs. 59.7 (88.4), p < 0.0001, CD19+CD27+, 42.1 (63.6) vs. 84.3 (78.1), p = 0.002, CD19+BAFF+, 59.7 (37.8) vs. 127.9 (123.7), p < 0.0001 and CD19+IgM+ cells, 48.9 (42.8) vs. 112.5 (81.7) (K/μL), p < 0.0001. The ratio of early/late apoptotic B lymphocytes was reduced (16.8 (10.9) vs. 110 (25.4), p = 0.03). CD19+CD5+ cells were the only cell type with an increased proportion in ESRD-T0 patients (2.7 (3.7) vs. 0.6 (1.1), p < 0.0001). After 6 months on CAPD or HD, CD19+CD27-(%) and early apoptotic lymphocytes were reduced further. The HD patients also showed a significant increase in late apoptotic lymphocytes, from 1.2 (5.7) to 4.2 (7.2) K/mL, p = 0.02. B cells and most of their subtypes were significantly reduced in ESRD-T0 patients compared to controls, the only exception being CD19+CD5+ cells. Apoptotic changes were prominent in ESRD-T0 patients and were exacerbated by HD.

Anticancer effect of antioxidant-rich methanolic extract of Rauvolfia serpentina (L.) Benth. ex Kurz leaves in HepG2 and HeLa cells: A mechanistic insight

Rauvolfia serpentina (L.) Benth. ex Kurz is a pharmaceutically, and agriculturally important medicinal plant traditionally used against snake and insect bites, dysentery, abdominal pain, fever, and insanity. However, no detailed study has been done on anticancer potency of crude leaf extract of Rauvolfia serpentina. Therefore, we investigated the anticancer activity of methanolic extract of Rauvolfia serpentina leaf (MERSL) in human hepatocarcinoma HepG2 and cervical cancer HeLa cells along with its antioxidant capacity. GC-MS analysis revealed the presence of many anticancer phytochemicals including campesterol, stigmasterol, beta-sitosterol, alpha-amyrin, lupeol, phytol, and D-pinitol. The antioxidant activity of MERSL was confirmed by DPPH (IC50 = 106.16 μg/ml), ABTS (IC50 = 146.67 μg/ml), FRAP (IC50 = 1.4 mg/ml), and total antioxidant capacity assay invitro, and reduction of H2O2-induced ROS in noncancer fibroblast F111 cells. Crystal violet assay showed that MERSL reduced cell viability in HepG2 (IC50 = 274.12 μg/ml) and HeLa (IC50 = 231.33 μg/ml) at 24 h, but not in F111 cells at these IC50 concentrations. Further, wound healing and clonogenic assays confirmed anticancer activity, which resulted in decreased cell migration and revival capacity in HepG2 and HeLa cells. Apoptotic changes were visualized by acridine orange-ethidium bromide and DAPI staining using fluorescence and confocal microscopy. Also, comet assay indicated DNA damage. An increase in intracellular ROS, calcium influx, reduction of mitochondrial mass and membrane potential, and cell cycle arrest, established by flow cytometry and fluorimetry, highlighted the possible underlying mechanism of cell death by MERSL in HepG2 and HeLa cells. Therefore, this study evidenced that Rauvolfia serpentina leaves could be an encouraging pharmacological application for developing anticancer strategies.

Combination of organic acids benzoate, butyrate, caprylate, and sorbate provides a novel antibiotics-independent treatment option in the combat of acute campylobacteriosis.

The food-borne Gram-negative bacterial pathogen Campylobacter jejuni may cause the acute enterocolitis syndrome campylobacteriosis in infected humans. Given that human C. jejuni infections are rising globally which hold also true for resistance rates against antibiotic compounds such as macrolides and fluoroquinolones frequently prescribed for the treatment of severe infectious enteritis, novel antibiotics-independent therapeutic strategies are needed. Distinct organic acids are well known for their health-beneficial including anti-microbial and immunomodulatory properties. In our present study, we investigated potential pathogen-lowering and anti-inflammatory effects of benzoic acid, butyric acid, caprylic acid, and sorbic acid either alone or in combination during acute murine campylobacteriosis. Therefore, secondary abiotic IL-10-/- mice were perorally infected with C. jejuni strain 81-176 and subjected to a 4-day-course of respective organic acid treatment. On day 6 post-infection, mice from the combination cohort displayed slightly lower pathogen loads in the duodenum, but neither in the stomach, ileum nor large intestine. Remarkably, the clinical outcome of C. jejuni induced acute enterocolitis was significantly improved after combined organic acid treatment when compared to the placebo control group. In support, the combinatory organic acid treatment dampened both, macroscopic and microscopic inflammatory sequelae of C. jejuni infection as indicated by less colonic shrinkage and less pronounced histopathological including apoptotic epithelial cell changes in the colon on day 6 post-infection. Furthermore, mice from the combination as compared to placebo cohort exhibited lower numbers of innate and adaptive immune cells such as neutrophilic granulocytes, macrophages, monocytes, and T lymphocytes in their colonic mucosa and lamina propria, respectively, which also held true for pro-inflammatory cytokine secretion in the large intestines and mesenteric lymph nodes. Notably, the anti-inflammatory effects were not restricted to the intestinal tract, but could also be observed systemically given pro-inflammatory mediator concentrations in C. jejuni infected mice from the combination organic acid treatment that were comparable to basal values. In conclusion, our in vivo study provides first evidence that an oral application of distinct organic acids in combination exhibits pronounced anti-inflammatory effects and hence, constitutes a promising novel antibiotics-independent therapeutic strategy in the combat of acute campylobacteriosis.