Investigation of Factors Affecting Extraction of Calophyllum inophyllum Seed Oil via Response Surface Methodology.

This study used the Soxhlet apparatus to investigate honne oil (HO) extraction optimization. Twenty-four (24) experiments were formulated using the D-optimal design considering extraction time (2 - 6 h), honne weight (20 - 60 g), and particle size using acetone. The yield, functional groups, physical and chemical properties, and fatty acid composition of the HO were assessed. The optimal extraction conditions established were a time of 6 h, fine particle size, and honne weight of 20 g with a high HO yield of 70.85 wt.%. The HO had an acid value and kinematic viscosity of 35.68 mg KOH/g oil and 52.96 mm 2 /s, respectively. The observed coefficient of determination of 0.9870 suggests that the model developed for the process is efficient. The functional groups and fatty acids of the HO confirm that it is highly unsaturated with the regions of trans-unsaturation bending vibrations and double bond stretching. The properties of the HO demonstrate that it could be used to produce biodiesel, notwithstanding the necessity for pretreatment.

Prognostic and Therapeutic Implications of Cell Division Cycle 20 Homolog in Breast Cancer.

Cell division cycle 20 homolog (CDC20) is a well-known regulator of cell cycle progression. Abnormal expression of CDC20 leads to mitotic defects, which play a significant role in cancer development. In breast cancer (BC), CDC20 has been identified as a biomarker that has been linked to poor patient outcomes. In this study, we investigated the association of CDC20 with BC prognosis and immune cell infiltration by using multiple online databases, including UALCAN, KM plotter, TIMER2.0, HPA, TNM-plot, bc-GenExMiner, LinkedOmics, STRING, and GEPIA. The results demonstrate that BC patients have an elevated CDC20 expression in tumor tissues compared with the adjacent normal tissue. In addition, BC patients with overexpressed CDC20 had a median survival of 63.6 months compared to 169.2 months in patients with low CDC20 expression. Prognostic analysis of the examined data indicated that elevated expression of CDC20 was associated with poor prognosis and a reduction of overall survival in BC patients. These findings were even more prevalent in chemoresistance triple-negative breast cancer (TNBC) patients. Furthermore, the Gene Set Enrichment Analysis tool indicated that CDC20 regulates BC cells' cell cycle and apoptosis. CDC20 also significantly correlates with increased infiltrating B cells, CD4+ T cells, neutrophils, and dendritic cells in BC. In conclusion, the findings of this study suggest that CDC20 may be involved in immunomodulating the tumor microenvironment and provide evidence that CDC20 inhibition may serve as a potential therapeutic approach for the treatment of BC patients. In addition, the data indicates that CDC20 can be a reliable prognostic biomarker for BC.

Non-cytotoxic 3-Acetylcoumarin as an Attractive Target for Human Monoamine Oxidase (HMAO) Enzymes.

BACKGROUND/AIM: Coumarins are a broad class of naturally occurring oxygen-heterocyclic compounds found in plants with diverse biological properties, making them attractive for evaluation as novel therapeutic agents. We herein report the in vitro cytotoxic and monoamine oxidase (MAO) inhibitory activities of 3-acetylcoumarins (6a-e). MATERIALS AND METHODS: The cytotoxic activity was evaluated using crystal violet dye binding assay, and those compounds unable to induce cytotoxicity were further tested for the monoamine oxidase (MAO) activity using the MAO-GloTM kit. RESULTS: The 3-acetylcoumarins (6a-e) were non-cytotoxic (inactive) against MDA MB-231 (estrogen receptor-negative, ER-, highly invasive) and MCF-7 (estrogen receptor-positive, ER+, weakly invasive) breast cancer cell lines, but showed interesting MAOs inhibition activities. Among the synthesized compounds, 3-acetylcoumarin bearing dichloro (-diCl) (6d; IC50=0.31±0.04 µM) at Carbon-7, 8 positions showed higher inhibition, MAO B/A non-selectivity (selectivity index, SI=3.10), reversible inhibition against the hMAO-B enzyme, and neuroprotection against H2O2-treated human neuroblastoma (N2a) cells. CONCLUSION: Compound (6d) can be considered a promising scaffold for further investigation in developing hMAO-B inhibitors (MAOIs).

Health Promotion and Racial Disparity in COVID-19 Mortality Among African American Populations.

COVID-19, known as Coronavirus Disease 2019, is a major health issue resulting from novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Its emergence has posed a significant menace to the global medical community and healthcare system across the world. Notably, on December 12, 2020, the Food and Drug Administration (FDA) approved the utilization of the Pfizer and Moderna COVID-19 vaccines. As of July 31, 2022, the United Stated has witnessed over 91.3 million cases of COVID-19 and nearly 1.03 million fatalities. An intriguing observation is the recent reduction in the mortality rate of COVID-19, attributed to an augmented focus on early detection, comprehensive screening, and widespread vaccination. Despite this positive trend in some demographics, it is noteworthy that the overall incidence rates of COVID-19 among African American and Hispanic populations have continued to escalate, even as mortality rates have decreased. Therefore, the objective of this research study is to present an overview of COVID-19, spotlighting the disparities among different racial and ethnic groups. It also delves into the management of COVID-19 within the minority populations. To reach our research objective, we used a publicly available COVID-19 dataset from kaggle:https://www.kaggle.com/datasets/paultimothymooney/covid19-cases-and-deaths-by-race. In addition, we obtained COVID-19 datasets from 10 different states with the highest proportion of African American populations. Many considerable strikes have been made in COVID-19. However, success rate of treatment in the African American population remains relatively limited when compared to other ethnic groups. Hence, there arises a pressing need for novel strategies and innovative approaches to not only encourage prevention measures against COVID-19, but also to increase survival rates, diminish mortality rates, and ultimately improve the health outcomes of ethnic and racial minorities.

Involvement of AKT/PI3K Pathway in Sanguinarine's Induced Apoptosis and Cell Cycle Arrest in Triple-negative Breast Cancer Cells.

BACKGROUND/AIM: Chemotherapy resistance in triple-negative breast cancer (TNBC) cells is well documented. Therefore, it is necessary to develop safer and more effective therapeutic agents to enhance the outcomes of chemotherapeutic agents. The natural alkaloid sanguinarine (SANG) has demonstrated therapeutic synergy when coupled with chemotherapeutic agents. SANG can also induce cell cycle arrest and trigger apoptosis in various cancer cells. MATERIALS AND METHODS: In this study, we investigated the molecular mechanism underlying SANG activity in MDA-MB-231 and MDA-MB-468 cells as two genetically different models of TNBC. We employed various assays including Alamar Blue to measure the effect of SANG on cell viability and proliferation rate, flow cytometry analysis to study the potential of the compound to induce apoptosis and cell cycle arrest, quantitative qRT PCR apoptosis array to measure the expression of different genes mediating apoptosis, and the western system was used to analyze the impact of the compound on AKT protein expression. RESULTS: SANG lowered cell viability and disrupted cell cycle progression in both cell lines. Furthermore, S-phase cell cycle arrest-mediated apoptosis was found to be the primary contributor to cell growth inhibition in MDA-MB-231 cells. SANG-treated TNBC cells showed significantly up-regulated mRNA expression of 18 genes associated with apoptosis, including eight TNF receptor superfamily (TNFRSF), three members of the BCL2 family, and two members of the caspase (CASP) family in MDA-MB-468 cells. In MDA-MB-231 cells, two members of the TNF superfamily and four members of the BCL2 family were affected. The western study data showed the inhibition of AKT protein expression in both cell lines concurrent with up-regulated BCL2L11 gene. Our results point to the AKT/PI3K signaling pathway as one of the key mechanisms behind SANG-induced cell cycle arrest and death. CONCLUSION: SANG shows anticancer properties and apoptosis-related gene expression changes in the two TNBC cell lines and suggests AKT/PI3K pathway implication in apoptosis induction and cell cycle arrest. Thus, we propose SANG's potential as a solitary or supplementary treatment agent against TNBC.

The Management of Diabetes Mellitus Using Medicinal Plants and Vitamins.

Diabetes mellitus (DM) is a serious chronic metabolic disease that is associated with hyperglycemia and several complications including cardiovascular disease and chronic kidney disease. DM is caused by high levels of blood sugar in the body associated with the disruption of insulin metabolism and homeostasis. Over time, DM can induce life-threatening health problems such as blindness, heart disease, kidney damage, and stroke. Although the cure of DM has improved over the past decades, its morbidity and mortality rates remain high. Hence, new therapeutic strategies are needed to overcome the burden of this disease. One such prevention and treatment strategy that is easily accessible to diabetic patients at low cost is the use of medicinal plants, vitamins, and essential elements. The research objective of this review article is to study DM and explore its treatment modalities based on medicinal plants and vitamins. To achieve our objective, we searched scientific databases of ongoing trials in PubMed Central, Medline databases, and Google Scholar websites. We also searched databases on World Health Organization International Clinical Trials Registry Platform to collect relevant papers. Results of numerous scientific investigations revealed that phytochemicals present in medicinal plants (Allium sativum, Momordica charantia, Hibiscus sabdariffa L., and Zingiber officinale) possess anti-hypoglycemic activities and show promise for the prevention and/or control of DM. Results also revealed that intake of vitamins C, D, E, or their combination improves the health of diabetes patients by reducing blood glucose, inflammation, lipid peroxidation, and blood pressure levels. However, very limited studies have addressed the health benefits of medicinal plants and vitamins as chemo-therapeutic/preventive agents for the management of DM. This review paper aims at addressing this knowledge gap by studying DM and highlighting the biomedical significance of the most potent medicinal plants and vitamins with hypoglycemic properties that show a great potential to prevent and/or treat DM.

Application of Machine Learning Algorithms in Breast Cancer Diagnosis and Classification.

Breast cancer continues to be the most frequent cancer in females, affecting about one in 8 women and causing the highest number of cancer-related deaths in females worldwide despite remarkable progress in early diagnosis, screening, and patient management. All breast lesions are not malignant, and all the benign lesions do not progress to cancer. However, the accuracy of diagnosis can be increased by a combination or preoperative tests such as physical examination, mammography, fine-needle aspiration cytology, and core needle biopsy. Despite some limitations, these procedures are more accurate, reliable, and acceptable, when compared with a single adopted diagnostic procedure. Recent studies have shown that breast cancer can be accurately predicted and diagnosed using machine learning (ML) technology. The objective of this study was to explore the application of ML approaches to classify breast cancer based on feature values generated from a digitized image of a fine-needle aspiration (FNA) of a breast mass. To achieve this objective, we used ML algorithms, collected a scientific dataset of 569 breast cancer patients from Kaggle (https://www.kaggle.com/uciml/breast-cancer-wisconsin-data), analyze and interpreted the data based on ten real-valued features of a breast mass FNA including the radius, texture, perimeter, area, smoothness, compactness, concavity, concave points, symmetry, and fractal dimension. Among the 569 patients tested, 63% were diagnosed with benign breast cancer and 37% were diagnosed with malignant breast cancer. Benign tumors grow slowly and do not spread while malignant tumors grow rapidly and spread to other parts of the body.

Pharmacological Effects of Selected Medicinal Plants and Vitamins Against COVID-19.

The coronavirus disease 2019 (COVID-19) is caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). It is a serious disease that has caused multiple deaths in various countries in the world. Globally, as of May 23, 2021, the total confirmed cases of COVID-19 have reach 166,346,635 with a total of 3,449,117 deaths. Several recent scientific studies have shown that medicinal plants and vitamins can benefit and improve the health of COVID-19 patients. However, the benefits of medicinal plants and vitamins in the treatment of COVID-19 remain unproven. Therefore, the objective of this article is to expounds the benefits of using medicinal plants (Allium sativum, curcumin, Nigella sativa, Zingiber officitale) and vitamins (vitamin C and vitamin D) that possess the antiviral properties for the prevention and/or control of COVID-19. To reach our objective, we searched scientific databases of ongoing trials in the Centers for Disease Control and Prevention websites, PubMed Central, Medline databases, and Google Scholar websites. We also searched databases on World Health Organization International Clinical Trials Registry Platform to collect relevant papers. We found that all of the selected medicinal plants and vitamins possess antiviral activities, and their individual intake shows promise for the prevention and/or control of COVID-19. We conclude that, the selected medicinal plants and vitamins possess anti-viral properties that are more likely to prevent and/or disrupt the SARS-CoV-2 replication cycle, enhance the human immune system and promote good health.

Chemo-Preventive Effect of Vegetables and Fruits Consumption on the COVID-19 Pandemic.

Coronavirus disease 2019 (COVID-19) is a new disease caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). It is a global pandemic that has claimed the death of 1,536,957 human beings worldwide including 287,842 deaths in the United States as of December 3, 2020. It has become a major threat to the medical community and the entire healthcare system in every part of the world. Recently, the Food and Drug Administration (FDA) has approved the emergency use of Pfizer and Moderna COVID-19 vaccine on December 12, 2020. However, there are concern about the new COVID-19 vaccine safety, efficacy, and immunity after the vaccination. In addition, both coronavirus and COVID-19 vaccine are new at this point and there is no scientific evidence to know whether people who are vaccinated can still carry the COVID 19 pathogens and pass them along to others. Therefore, many people all over the world have an increased interest in consuming more VF for the purpose of maintaining their health and boosting their immune system. Identifying novel antiviral agents for COVID-19 is of critical importance, and VF is an excellent source for drug discovery and therapeutic development. The objective of this study is to test the hypothesis that a high intake of vegetables and/or fruits prevents COVID-19 incidence and reduces the mortality rate. To achieve this objective, we collected the diet data of COVID-19 from Kaggle (https://www.kaggle.com/mariaren/covid19-healthy-diet-dataset), and used a machine-learning algorithm to examine the effects of different food types on COVID-19 incidences and deaths. Specifically, we used the feature selection method to identify the factors (e.g., diet-related factors) that contribute to COVID-19 morbidity and mortality. Data generated from the study demonstrated that VF intake can help to combat the SARS-CoV-2. Taken together, VF may be potential chemopreventive agents for COVID-19 due to their antiviral properties and their ability to boost the human body immune system.

Evaluation of microbial diversity of three recreational water bodies using 16S rRNA metagenomic approach.

Surface water plays a significant role in world development by promoting economic growth and health benefits to humans and animals whose lives depend on good water quality in the ecosystem. Thus, this study investigated the differences in physical and chemical properties of surface water from two lakes (Lakes Jackson and Talquin) and a pond (Pedrick Pond). Also, the influence of environmental factors on the microbial communities that live within the water environment was examined. Genomic DNA was extracted from the water samples collected and 16S rRNA sequencing method was employed to characterize the microbial community compositions across the three locations. The results obtained suggest that the water sources met the recommended recreational water quality criteria standard for clean water. Overall, Proteobacteria, Actinobacteria, Cyanobacteria, Bacteroidetes were the main bacterial phyla present in the communities, while Archaea was mainly dominated by Euryachaeota. Pressure, conductivity, temperature, dissolved oxygen (DO), and pH accounted for 74.2% of the variation in the distribution of the microbial community in the three locations (P < 0.05), while 58.2% of the variation in the microbial community distribution was accounted for by pressure and conductivity. The high temperature observed in the Pedrick Pond correlated with the distribution of genera hgcl_clades and Legionella. While in Lake Talquin, water conductivity was significantly associated with the abundance of Cyanobium_PCC_6307, Sediminibacterium, and Conexibacter. The results from this study indicate that the microbial communities in the two lakes are different from the pond and all the environmental variables accounted for a significant portion of the total variation, but pressure, conductivity, and temperature are more important factors due to significant correlation with the distribution of the microbial communities.

Identification of 7,8-dihydroxy-3-phenylcoumarin as a reversible monoamine oxidase enzyme inhibitor.

We herein report the biological evaluation of 3-arylcoumarin derivatives (3a-l) as potential human monoamine oxidase-A and -B (hMAO-A and hMAO-B) inhibitors. The result indicated that 7,8-dihydroxy-3-(4-nitrophenyl)coumarin (3j) was most effective against MAO-A (inhibition concentration [IC50 ] = 6.46 ± 0.02 µM) and MAO-B (IC50 = 3.8 ± 0.3 µM) enzymes than other synthesized compounds and reference compounds (pargyline and moclobemide). Furthermore, compound (3j) showed (a) nonselectivity against hMAO enzymes, (b) reversible hMAO enzymes inhibition, and (c) neuroprotection against H2 O2 -treated human neuroblastoma (N2a) cells. Finally, a molecular modeling study revealed that the hMAO enzymes inhibitory activity of the compound (3j) may be due to the orientation where the nitro (NO2 ) group lies deep into the receptor and the phenyl ring directed toward flavin adenosine dinucleotide via hydrogen bond interaction, and possible π-π interaction with various important residues. Thus, the results of the present study demonstrate that compound (3j) can be considered as a promising scaffold for the development of hMAO-A and hMAO-B inhibitors.

Identification of cytotoxic markers in methamphetamine treated rat C6 astroglia-like cells.

Methamphetamine (METH) is a powerfully addictive psychostimulant that has a pronounced effect on the central nervous system (CNS). The present study aimed to assess METH toxicity in differentiated C6 astroglia-like cells through biochemical and toxicity markers with acute (1 h) and chronic (48 h) treatments. In the absence of external stimulants, cellular differentiation of neuronal morphology was achieved through reduced serum (2.5%) in the medium. The cells displayed branched neurite-like processes with extensive intercellular connections. Results indicated that acute METH treatment neither altered the cell morphology nor killed the cells, which echoed with lack of consequence on reactive oxygen species (ROS), nitric oxide (NO) or inhibition of any cell cycle phases except induction of cytoplasmic vacuoles. On the other hand, chronic treatment at 1 mM or above destroyed the neurite-like processors and decreased the cell viability that paralleled with increased levels of ROS, lipid peroxidation and lactate, depletion in glutathione (GSH) level and inhibition at G0/G1 phase of cell cycle, leading to apoptosis. Pre-treatment of cells with N-acetyl cysteine (NAC, 2.5 mM for 1 h) followed by METH co-treatment for 48 h rescued the cells completely from toxicity by decreasing ROS through increased GSH. Our results provide evidence that increased ROS and GSH depletion underlie the cytotoxic effects of METH in the cells. Since loss in neurite connections and intracellular changes can lead to psychiatric illnesses in drug users, the evidence that we show in our study suggests that these are also contributing factors for psychiatric-illnesses in METH addicts.

Diallyl disulfide attenuation effect on transcriptome in rat liver cells against cadmium chloride toxicity.

In this report, liver cells were treated with cadmium chloride (CdCl2 ) and diallyl disulfide (DADS), a major compound from garlic to attenuate the toxic effect of Cd on transcriptome. The viability of Cd treated cells was reduced to 19.9% ± 2.4% in comparison to the untreated cells, whereas the viability of DADS pretreated cells was increased to 48.6% ± 2%. The attenuation effect of DADS was studied at shorter period (6 hours). Transcriptome analysis of CdCl2 alone treated cells resulted in 2119 and 982 (up and down) regulated genes (≥ 2 or ≤ 2-fold), whereas pretreated cells with DADS resulted in 2597 and 1784 genes. These genes were known to function in many important biological processes. Affymetrix array analysis was validated by the pathway specific PCR array that exhibited the same trend of expression. The current study clearly shows the DADS attenuation effect on transcriptome in CdCl2 -treated rat liver cells.

7,8-Dihydroxy-3-arylcoumarin Induces Cell Death Through S-Phase Arrest in MDA-MB-231 Breast Cancer Cells.

BACKGROUND/AIM: Coumarins remain one of the most versatile classes of compounds for anticancer drug design and discovery. The present study aimed to evaluate the in vitro cytotoxic activity of 7,8-Dihydroxy-3-arylcoumarin derivatives (7a-i) in A549, MDA-MB-231and PC-3 cancer cell lines. MATERIALS AND METHODS: Cell viability, cell-cycle progression and regulatory protein expression were evaluated using crystal violet dye-binding assay, flow cytometry and western blot analysis. RESULTS: 7,8-Diacetoxy-3-(4-nitrophenyl)coumarin (7h) showed the highest cytotoxic activity with CC50 of 7.51±0.07 µM in MDA-MB-231 cell line. The mechanism of cytotoxic action indicated that 7h caused significant (p<0.05) MDA-MB-231 cells arrest in the S phase as well as moderate cells arrest in the G2/M phase; confirmed by up-regulation of cyclins A/B1, p21 and CDKs 4/6, and down-regulation of cyclin E2 and CDK2 regulatory proteins. CONCLUSION: These results suggest that 7h could serve as a valuable template for the development of novel synthetic compounds for breast cancer treatment.

7,8-Dihydroxy-3-(4-nitrophenyl)coumarin induces cell death via reactive oxygen species-independent S-phase cell arrest.

We herein report the synthesis and in vitro cytotoxicity of 3-arylcoumarin derivatives (6a-f and 7a-f) in human liver (HepG2), prostate (LNCap), and pancreatic (BxPC3) cancer cell lines. Among the tested compounds, 7,8-dihydroxy-3-(4-nitrophenyl) coumarin (7b) showed the highest cytotoxicity in the HepG2 cell line. The mechanism of cytotoxic action indicated that compound (7b) arrested HepG2 cells at the S phase of the cell cycle progression, induced loss of mitochondrial membrane potential, and caused reactive oxygen species (ROS)-independent cell death. The cell viability result of pretreated HepG2 cells with antioxidant N-acetylcysteine followed by compound (7b) treatment and the free radical scavenging activities of compound (7b) confirmed the ROS-independent cell death. These results demonstrate that compound (7b) could serve as a valuable template for the development of novel synthetic compounds as potential anticancer agents for hepatocellular carcinoma treatment.

Cellular and molecular responses to acute cocaine treatment in neuronal-like N2a cells: potential mechanism for its resistance in cell death.

Cocaine is a highly abused drug that causes psychiatric and neurological problems. Its entry into neurons could alter cell-biochemistry and contribute in the manifestation of early pathological symptoms. We have previously shown the acute cocaine effects in rat C6 astroglia-like cells and found that these cells were highly sensitive to cocaine in terms of manifesting certain pathologies known to underlie psychological disorders. The present study was aimed to discern acute cocaine effects on the early onset of various changes in Neuro-2a (N2a) cells. Whole-cell patch-clamp recording of differentiated cells displayed the functional voltage-gated Na+ and K+ channels, which demonstrated the neuronal characteristics of the cells. Treatment of these cells with acute cocaine (1 h) at in vivo (nM to µM) and in vitro (mM) concentrations revealed that the cells remained almost 100% viable. Cocaine administration at 6.25 µM or 4 mM doses significantly reduced the inward currents but had no significant effect on outward currents, indicating the Na+ channel-blocking activity of cocaine. While no morphological change was observed at in vivo doses, treatment at in vitro doses altered the morphology, damaged the neurites, and induced cytoplasmic vacuoles; furthermore, general mitochondrial activity and membrane potential were significantly decreased. Mitochondrial dysfunction enabled the cells switch to anaerobic glycolysis, evidenced by dose-dependent increases in lactate and H2S, resulting unaltered ATP level in the cells. Further investigation on the mechanism of action unfolded that the cell's resistance to cocaine was through the activation of nuclear factor E2-related factor-2 (Nrf-2) gene and subsequent increase of antioxidants (glutathione [GSH], catalase and GSH peroxidase [GPx]). The data clearly indicate that the cells employed a detoxifying strategy against cocaine. On a broader perspective, we envision that extrapolating the knowledge of neuronal resistance to central nervous system (CNS) diseases could delay their onset or progression.

Comparative whole genome transcriptome analysis and fenugreek leaf extract modulation on cadmium­induced toxicity in liver cells.

Cadmium (Cd), an economically valuable metal, is widely used in various industrial processes. Although it is of economic value, it is hazardous to human health. Cd accumulates in vital organs where it causes various diseases. Natural compounds with chelating or antioxidant properties have been tested to reduce the toxic effect of Cd. The anti­oxidant, anti­diabetic and hypocholesterolemic properties of fenugreek (Trigonella foenum-graecum) leaves make it a candidate for investigation as protective agent against Cd­induced toxicity. In the present study, the protective effects of fenugreek leaf extract (FLE) on cell viability, morphology, and whole genomic transcription in cadmium chloride (CdCl2)­treated rat liver cells were analyzed. The cells were treated with 25 µM CdCl2 alone, or co­treated with 5 µg/ml FLE for 48 h. The co­treated cells were pretreated with FLE for 2 or 4 h, followed by CdCl2 treatment. Genomic transcription analysis was performed in the CdCl2­treated cells following treatment for 6 h. The CdCl2 caused a significant decrease in viability (35.8±4.1%) and morphological distortion of the cells, compared with the untreated control cells; whereas 4 h pretreatment with FLE (5 µg/ml) reversed the Cd­induced morphology alteration and increased the cell viability to 102±3.8%. Genomic transcription analysis of the CdCl2 only­treated cells showed 61 upregulated and 124 downregulated genes, compared with 180 upregulated and 162 downregulated genes in the FLE pretreated cells. Furthermore, 37 and 26% of the affected total genomic genes in the CdCl2 only­treated cells were involved in binding and catalytic activities, respectively, whereas 50 and 20% of the genes in the FLE pretreated cells were involved in binding and catalytic activities, respectively. In conclusion, these results suggested that genome transcriptome modulation may be important in the protective effect of FLE against Cd­induced toxicity in normal rat liver cells.

Identification of 7,8-Diacetoxy-3-Arylcoumarin Derivative as a Selective Cytotoxic and Apoptosis-inducing Agent in a Human Prostate Cancer Cell Line.

BACKGROUND/AIM: Coumarins are a member of the benzopyrone family of compounds with diverse and interesting pharmacological properties. In the present study, we report the in vitro cytotoxicity evaluation of 7,8-Diacetoxy-3-arylcoumarin derivatives (5a-h) in human prostate (PC-3) and breast (MDA-MB-231) cancer cell lines. MATERIALS AND METHODS: The cytotoxic activity was evaluated using crystal violet dye-binding assay. Furthermore, the most active compound in vitro cytotoxic activity in human non-cancerous cell line and its effect on the cell-cycle phases, apoptosis proteins expression, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) production and Glutathione (GSH) level were performed. RESULTS: Among the eight compounds that were evaluated, 7,8-Diacetoxy-3-(4-methylsulfonyl phenyl)coumarin (5f) was the most active derivative with highest cytotoxic activity and selectivity against the PC-3 cell line vs. the non-cancerous WPE1-N22 cell line. The cytotoxic action of compound 5f in PC-3 cells is associated with the cell-cycle arrest at -G0/G1 phase, apoptosis, loss in mitochondrial membrane potential (MMP), induced reactive oxygen species (ROS) production and depletion of Glutathione (GSH) level. CONCLUSION: The result indicates that the presence of p-methylsulfonylphenyl group on compound 5f is critical in modulating selective cytotoxic activity and induction of apoptosis via the mitochondrial apoptotic signaling pathway that is independent of cytochrome c release.

The antiangiogenic effects of polyisoprenylated cysteinyl amide inhibitors in HUVEC, chick embryo and zebrafish is dependent on the polyisoprenyl moiety.

Angiogenesis is essential for solid tumor growth, therapeutic resistance and metastasis, the latest accounting for 90% of cancer deaths. Although angiogenesis is essential for the malignant transformations in solid tumors and therefore is an attractive target, few drugs are available that block tumor angiogenesis. The focus has been to block signaling by receptor tyrosine kinases (RTKs), such as for vascular endothelial growth factor (VEGF), whose activation abrogate apoptosis and promote angiogenesis. The polyisoprenylated cysteinyl amide inhibitors (PCAIs) were designed to modulate aberrant polyisoprenylated small G-proteins such as mutant Ras whose constitutive activation promotes RTKs signaling. Since polyisoprenylation is essential for protein-protein interactions and functions of G-proteins, we hypothesized that the PCAIs would disrupt the monomeric G-protein signaling thereby effectively inhibiting angiogenesis. In this study we determined the effects of PCAIs on human umbilical vein endothelial cells (HUVEC) tube formation, cell viability, cell migration and invasion as well as in vivo using the chick chorioallantoic membrane (CAM) and zebrafish models. At sub- to low micromolar concentrations, the PCAIs inhibit the native and VEGF-stimulated cell migration and invasion as well as tube formation and angiogenesis in CAM and zebrafish embryos. The concentrations that block the angiogenic processes were lower than those that induce cell death. Since angiogenesis is essential for tumor growth but otherwise limited to wound healing, feeding fat cells and uterine wall repair in adults, it is conceivable that these compounds can be developed into safer therapeutics for cancers and retinal neovascularization that leads to loss of vision.

Modulation of cytokines and chemokines expression by NAC in cadmium chloride treated human lung cells.

Cadmium (Cd), is one of the most hazardous metals found in the environment. Cd exposure through inhalation has been linked to various diseases in lungs. It was shown that Cd induces proinflammatory cytokines through oxidative stress mechanism. In this report, we studied the immunomodulatory effect of a well known antioxidant, N-acetylcysteine (NAC) on cadmium chloride (CdCl2 ) treated human lung A549 cells through human cytokine array 6. The lung cells were treated with 0 or 75 µM CdCl2 alone, 2.5 mM NAC alone, or co-treated with 2.5 mM NAC and 75 µM CdCl2 for 24 h. The viability of cells was measured by crystal violet dye. The array results were validated by human IL-1alpha enzyme- linked immunosorbent assay (ELISA) kit. The viability of the 75 µM CdCl2 alone treated cells was decreased to 44.5%, while the viability of the co-treated cells with 2.5 mM NAC was increased to 84.1% in comparison with untreated cells. In the cell lysate of CdCl2 alone treated cells, 19 and 8 cytokines were up and down-regulated, while in the medium 15 and 3 cytokines were up and downregulated in comparison with the untreated cells. In the co-treated cells, all these cytokines expression was modulated by the NAC treatment. The IL-1α ELISA result showed the same pattern of cytokine expression as the cytokine array. This study clearly showed the modulatory effect of NAC on cytokines and chemokines expression in CdCl2- treated cells and suggests the use of NAC as protective agent against cadmium toxicity. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1612-1619, 2016.