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Traditional Palestinian medicinal plant Cercis siliquastrum (Judas tree) inhibits the DNA cell cycle of breast cancer – Antimicrobial and antioxidant characteristics
Abstract
Introduction: Cercis siliquastrum (Judas tree) is a traditional medicine with drug discovery potential. The aim of this study was to investigate the antimicrobial and antioxidant activity of Cercis siliquastrum L. flowers and leaves for their effect on the DNA cell cycle (proliferation) in a breast cancer cell line.
Methods: Assessment of the DNA cell cycle of breast cancer was conducted using propidiumiodide (PI) while apoptosis activity was estimated by staining with Annexin-V using flow cytometry. The antimicrobial activity was assessed through a broth microdilution method on four bacterial pathogens and one fungal strain. Moreover, the Diphenylpicrylhydrazyl (DPPH) method was utilized to evaluate free radical scavenging efficacy of C. siliquastrum flowers and leaves in four different solvent fractions.
Results: The acetone and methanol fractions of C. siliquastrum leaves and flowers showed highest antioxidant potentials with IC50 values of 8.31 ± 1.36, 4.78 ± 1.84, 1.75 ± 2.03 and 3.31 ± 1.66 μg/ml, respectively compared with Trolox that had an antioxidant potential of IC50 value of 1.41 ± 1.05 μg/ml. The leaf hexane fraction showed potent antibacterial potential versus Staphylococcus aureus, Methicillin-Resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa with MIC values of 0.007, 0.024, 0.048 mg/ml, respectively. Notably, the C. siliquastrum flowers hexane fraction strongly inhibited growth of Staphylococcus aureus, Pseudomonas aeruginosa, MRSA, and Candida albicans with MIC values of 0.009, 0.039, 0.048 and 0.08 mg/ml, respectively. Moreover, C. siliquastrum leaves and flowers hexane fractions caused cell cycle arrest followed by cell death via apoptosis/necrosis due predominantly to defects in the mitotic process.
Conclusion: Our data revealed the significant antimicrobial and antioxidant effects of C. siliquastrum that contributed significantly to the cytotoxicity of cancer cells requires further detailed studies.
... Cercis siliquastrum, commonly known as the Judas tree, is a small perennial tree with pink flowers and heartshaped dark green leaves. Cercis siliquastrum can be used in traditional medical applications as an antiseptic, digestive cure, and medicine against infectious diseases, as well as for decoration purposes [22]. It is native to southeastern Europe and southern and western Asia, in addition, it is widely found in continental Greece, the northern Aegean, and the northern Ionian Islands. ...
Cercis siliquastrum seeds were used for the adsorption of the basic dyes namely basic blue 9 and basic green 4 from the wastewater. Adsorption experiments were conducted under various conditions including different temperatures (25, 35, and 45 °C), pH (2–12), contact time (0–180 min), adsorbent amount (0.2–4.0 g/L), and concentration (10–50 mg/L). The adsorption processes were applied to various isotherms such as Freundlich, Langmuir, Dubinin-Radushkevich, and Harkins–Jura. Furthermore, different adsorption kinetic models including pseudo-first-order, pseudo-second-order, Boyd, and intraparticle diffusion were studied to comprehend the mechanism of the adsorption of the dyes. The results illustrated that the adsorption was best described by the Langmuir (r² = 0.9949 for basic blue 9 dye and r² = 0.9939 for basic green 4 dye at 25 °C). From the Langmuir isotherm, the qm value of basic blue 9 was found as 500.0 mg/g whereas those of basic green 4 was found as 243.9 mg/g at 25 °C. According to the thermodynamic parameters, it was determined that the adsorption process occurred endothermically and spontaneously for basic blue 9 and basic green 4 dyes. Considering the capacity of adsorption for both dyes, this study suggests that Cercis siliquastrum seeds are low-cost and environmentally friendly materials for the effective adsorption of basic dyes.
Graphical abstract
... These tests are used to evaluate the coagulation cascade, PT is used for evaluation of the extrinsic and common pathways of the coagulation cascade. APTT for the intrinsic and common pathways, and TT for the conversion of brinogen to brin in the common pathway (17,18). The results suggest that the inhibition of clotting factors in the intrinsic and common pathways because both aPTT and TT tests were prolonged signi cantly. ...
Background: Wild plants are amply utilized in traditional medicine and folkloric food worldwide. Arum palaestinum Boiss. (AP) is one of the wild Palestinian plants which leaves have a long history in the Middle Eastern countries as food and medicine. Herby, the current study aimed to evaluate the antimicrobial, coagulation cascade activities, and anticancer effects of (AP) flowers extract
Methods: The aqueous extract of (AP) flowers was screened on its antimicrobial activity using microdilution assay against eight pathogens. While, prothrombin time, activated partial thromboplastin time, and thrombin time tests were measured utilizing standard hematological methods. And Anti cancer effect was assessed by using Parameters of cell cycles and alph feta protein level that were investigated for (AP) flowers fractionated with aqueous, DMSO, and methanol
Results: The antimicrobial screening results revealed that the aqueous extract of (AP) has strong antibacterial effects against P. vulgaris and E. faecium compared with Ampicillin with MIC values of 6.25, 6.25 and 18 mg/ml, respectively. The aqueous extract of (AP) showed anticoagulant activity with significant prolonged results in aPTT and TT tests at high concentrations (50 mg/ml and 25 mg/ml) and slightly prolonged results in the PT test at a high concentration (50 mg/ml).
The anticancer results indicate a delay in cell cycle through decreased the cell proliferation rate following effects of the AP fractions. The delay in the S phase was in favor of the water fraction. Water and DMSO fractions while maintained the cells in the G2-M phase similar to the DOX, the flower extract in methanol accelerated the cells in the G2-M phase suggesting that (AF) flower extracts have anti-cancer properties. At the same time Aqueous extract decreased HCC aFP to 1.55-fold (P=0.0008). While DMSO and methanolic extract had no significant effects on HCC aFP levels, compared to control untreated cells of 2519.16 ± 198.1 ng/ml. This data show that (AF) aqueous solution is potent inhibitor of alpha-fetoprotein secretion (P-value <0.05), which indicates its anti-carcinogenic effects
Conclusion: These results showed that the aqueous extract of (AP) plant possesses bioactive components with antibacterial and anticoagulant properties, which may be exploited in the treatment of infectious diseases and blood coagulation disorders.
... They are used in Europe and Middle East to enrich salads, fried to produce omelettes or preserved in vinegar. The flowers extract shows antioxidants and antimicrobial activity (Amer et al., 2019) Clitoria ternatea (butterfly pea) is a perennial climbing with plant woody foot originally from South East Asia. The beautiful blue flowers are very famous and are used to decorate recipes or to prepare blue teas and blue coloured dishes. ...
... For thousands of years, people tried to cure their diseases by utilizing various available natural materials and among these were medicinal plants [1,2]. Recently, it is well recognized that diabetes mellitus is considered one of the most common diseases worldwide, which is characterized as a type of metabolic disorder known by hyperglycemia, hypoinsulinemia, hyperlipidemia, and increased oxidative stress [3]. ...
... Nevertheless, the folkloric remedies have always been known as a rich source of phytochemicals which are important for the discovery of potent new medications especially these compounds that are working on communicable diseases [22]. ...
Pellitory plant (Parietaria judaica (PJ)) is one of the most widely used Arabian traditional medicinal plants due to its ability to cure several infectious diseases and other illnesses. The current study is aimed at assessing the phytoconstituents, antilipase, antiamylase, antimicrobial, and cytotoxic characters of the Pellitory plant (Parietaria judaica (PJ)). Phytochemical screening and procyanidin detection were conducted according to the standard phytochemical procedures. Porcine pancreatic lipase and α-amylase inhibitory activities were carried out using p-nitrophenyl butyrate and dinitrosalicylic acid assays, respectively. In addition, antimicrobial activity was determined utilizing a microdilution assay against several bacterial and fungal strains. Besides, the cytotoxic effect against HeLa cell line was tested employing 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. The quantitative test results revealed that the methanol fraction of PJ contains mg of procyanidin and has a potential α-amylase inhibitory activity compared with the antidiabetic drug Acarbose with IC50 values of and μg/ml, respectively. Also, it has a potential antilipase activity compared to the commercial antiobesity drug, Orlistat, with IC50 values of and μg/ml, respectively. The acetone, hexane, and methanol fractions have broad-spectrum antibacterial activity against the screened bacterial strains, while the acetone fraction has shown anticandidal activity with a MIC value of 0.195 mg/ml. The PJ hexane and acetone fractions decreased HeLa cell viability significantly ( value < 0.0001) by approximately 90% at the concentration of 0.625 mg/ml. The revealed outcomes showed that the methanol fraction has strong α-amylase and lipase inhibitory characters. Besides, acetone, hexane, and methanol fractions have broad-spectrum antibacterial activity, while the acetone fraction revealed potent antifungal activity against Candida albicans. Moreover, at low concentrations, hexane and acetone fractions have potent cytotoxic and antiproliferative activity against HeLa cancer cells. Nevertheless, PJ acetone, hexane, and methanol fractions can serve as an effective source of natural products to develop new antiobesity, antidiabetic, antimicrobial, and anticancer agents.
1. Introduction
Humans, since time immemorial, have relied on herbs and other natural products for the recovery and prophylaxis of many illnesses. In addition to their effective potentials in the therapeutic features, herbal products are easier to obtain, less expensive, and more acceptable for people than synthetic medicines. However, herbs have risks and sometimes can lead to poisoning if not used properly, and some of them are inefficient in the treatment of certain emergency cases [1].
There is a worldwide epidemic of overweight and obesity, which are usually associated with several pathologies such as diabetes mellitus, cardiovascular diseases, musculoskeletal disorders, and some types of cancer including colon, kidney, gallbladder, prostate, liver, ovarian, breast, and endometrial [2].
Diabetes is known by a long term of hyperglycemia with disturbances in the metabolism of proteins, fats, and carbohydrates, which resulted from defects in insulin action and/or insulin secretion. The main goal of any antidiabetic medicines is to reach normoglycemia to prevent microvascular and macrovascular complications [3].
Bacterial infections are considered a worldwide problem and are recognized as a threat to the life of humankind. In recent years, antibacterial and antifungal resistance has become an emergent issue in health worldwide. This resistance is mainly caused by the misuse of antibiotics [4].
According to the World Health Organization (WHO) surveys, cancer is one of the leading causes of death around the globe and responsible for about 10 million deaths in 2018. About 1 out of 6 people died from cancer which is considered the hugest cause of death, which is a considerably alarming estimate. The WHO has recognized that 1.16 trillion US dollars were spent on the prevention and treatment of cancer in 2010 alone, and that number has increased dramatically over the years [5].
Parietaria judaica L. (PJ) is commonly known as pellitory which belongs to the Urticaceae family and wildly growing in the northern countries of Africa, western regions of Asia, and the southern parts of Europe [6]. It is a perennial upright or spreading herbaceous plant reaching 1 m in height. The stems are greenish-brown or reddish-brown, are often much-branched, and are covered in irregularly curled hairs. The leaves have an oval shape, which is covered in irregular hairs, and they also have glossy upper surfaces. The flowers are borne in small, dense clusters in the leaf forks. They are initially greenish, but often turn reddish or reddish-brown as they mature [7].
The aqueous and lipophilic solutions prepared from PJ leaves have been broadly exploited for its medicinal value for centuries. This herb is used in traditional medicine for the treatment of kidney and bladder stones and to remove plaques deposited on the teeth. Furthermore, it is used for more thousand years as a diuretic and sedative as well as for the treatment of chronic cough, inflamed wounds, and burns [8, 9].
The current investigation is aimed at finding out more about the phytoconstituents and total tannin constituent of PJ and at investigating its antimicrobial effects against eight lethal microbial strains. Moreover, the plant antiobesity, antidiabetic, and cytotoxic characters against the HeLa cancer cell line were studied.
2. Material and Methods
2.1. Collection of the Plant Material
The leaves of PJ were gathered in November 2018 from the Jenin area of Palestine. The plant was recognized by Dr. Nidal Jaradat specialist in medicinal plants, and the voucher specimen was deposited in the Pharmacognosy Laboratory, Faculty of Medicine and Health Sciences at An-Najah National University (Pharm-PCT-1790).
The leaves were washed three times using distilled water and completely dried in the shade at room temperature. The dried parts were grounded coarsely using a mini mill machine and latterly stored in tightly sealed special containers for further use.
2.2. Four Solvent Exhaustive Fractionations
The dried leaves were exhaustively extracted by the fractionation method utilizing four solvents with various degrees of polarities including methanol (Loba/Chemie, India), water, acetone (Riedel/dehaen, Germany), and hexane (Alfa۔Aesar, UK). Briefly, 100 g of the dried plant leaves was taken and placed in a bottle and then extracted with 1 L of each solvent separately. Each bottle containing the plant leaves and the solvents was soaked for 72 h in a shaker device (Daihan Labtech, S. Korea) at 100 rotations per minute at 25°C. Each solvent was filtered utilizing a suction filtration. Then, all the organic fractions were dried using an incubator device (Esco, 2012-74317, Singapore) at 25°C until completely dried. A freeze dryer (Mell rock, China) was used in the drying of water fraction. Each obtained dried fraction was stored in the refrigerator at a temperature of 2-8°C for later use (5).
2.3. Phytochemical Screening
It is well known that plants produce many organic chemical compounds that are biologically active, not just in themselves, but also in other organisms. Some of these chemicals enhance the plants’ survival. Preliminary phytochemical analysis of secondary and primary metabolic compounds such as cardiac glycosides, flavonoids, saponins, proteins, phenols, carbohydrates, and tannins was carried out according to the standard phytochemical methods [10, 11].
2.4. Procyanidin Determination
For the determination of total procyanidin content, the Sun et al. protocol was followed with minor modification [12]. Catechin (Sigma, USA) was used as the reference compound to construct the calibration curve for the required calculations in which a 100 μg/ml stock methanolic solution was prepared; then, serial dilutions were obtained (10, 30, 50, 70, and 100 μg/ml). Then, a 4% methanolic vanillin (Alfa۔Aesar, UK) solution was freshly prepared, and a 100 μg/mL stock solution was made from the methanolic plant fraction using methanol as the solvent. For the working solution, each test tube contained 0.5 ml of the PJ plant methanolic fraction mixed with 3 ml of vanillin solution and 1.5 ml of concentrated HCl (SDFCL, India). The obtained mixture was allowed to stand for 15 min, and then, the absorption was measured at 500 nm against methanolic vanillin as a blank. All the working samples were analyzed in triplicate. The total procyanidin content in the plant fraction is expressed as Catechin equivalents (mg of CAE/g of the dry plant fraction).
2.5. Porcine Pancreatic Lipase Inhibitory Assay
The porcine pancreatic lipase inhibitory method was followed in this study according to the protocol of Bustanji et al., with minor modifications [13]. Briefly, a stock solution of 500 μg/ml from each plant fraction was dissolved in 10% DMSO which was used to prepare five different solutions with the following concentrations: 50, 100, 200, 300, and 400 μg/ml. One mg/ml stock solution of pancreatic lipase enzyme was obtained which is an enzyme that breaks down triglycerides into free fatty acids and glycerol. It is present in pancreatic secretions and is responsible for fat digestion and plays a crucial role in lipid transport. This enzyme was freshly prepared in the tris-HCl buffer before use. The substrate used for this study, p-nitrophenyl butyrate (PNPB), was prepared by dissolving 20.9 mg in 2 ml of acetonitrile. For each working test tube, 0.1 ml of porcine pancreatic lipase (1 mg/ml) was mixed with 0.2 ml of each diluted solution series for each plant fraction. The resulting mixture was then brought to a total volume of 1 ml, by adding a Tris-HCl solution and incubated at 37°C for 15 min. Following the incubation period, 0.1 ml of PNPB solution was added to each test tube. The mixture was incubated for 30 min at 37°C. Antilipase activity of PJ plant four solvent fractions was determined by measuring the hydrolysis of the PNPB compound into p-nitrophenolate ions at 410 nm using a UV-Visible spectrophotometer. The same procedure was repeated for Orlistat, which was used as a standard reference compound. The equation used in this analytical study is shown below: is the recorded absorbance of the blank solution and is the recorded absorbance of the (PJ) sample solution.
2.6. α-Amylase Inhibitory Activity
The α-amylase inhibitory activity of each extract fraction was carried out according to the standard method, with minor modifications [14]. Each plant fraction was dissolved in 3 ml of 10% DMSO and then further dissolved in buffer (0.02 M of Na2HPO4/NaH2PO4, 0.006 M NaCl, at pH 6.9) to give concentrations of 1000 μg/ml, from which the following dilutions were prepared: 10, 50, 70, 100, and 500 μg/ml. The porcine pancreatic α-amylase enzyme solution was freshly prepared at a concentration of 2 units/ml in 10% DMSO.
For working solutions, a volume of 0.2 ml of enzyme solution was mixed with 0.2 ml of each (PJ) fraction and was incubated for 10 min at 30°C. After the incubation period, 0.2 ml of a freshly prepared 1% starch aqueous solution was added to each working solution, followed by an incubation period of at least 3 min. The reaction was quenched by the addition of 0.2 ml dinitrosalicylic acid (DNSA) yellow color reagent. Each working solution was then diluted with 5 ml of distilled water and then boiled for 10 min in a water bath at 90°C. The mixture was cooled to room temperature, and the absorbance was taken at 540 nm. The blank was prepared following the same steps above, but the plant fraction was replaced with 0.2 ml of the previously described buffer. Acarbose was used as the standard reference following the same steps used for plant fractions.
The α-amylase inhibitory activity was calculated using the following equation: where is the absorbance of blank and is the absorbance of (PJ) sample.
2.7. Antimicrobial Activity
The antibacterial effect was determined using seven strains of bacteria which were brought from the American Type Culture Collection (ATCC): Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 25922), Klebsiella pneumonia, (ATCC 13883), Proteus vulgaris (ATCC 8427), Enterococcus faecium (ATCC 700221), and Staphylococcus aureus (ATCC 25923) as well as against the growth of a diagnostically confirmed Methicillin-Resistant Staphylococcus aureus (MRSA). The antifungal activity of (PJ) samples was evaluated against the growth of Candida albicans (ATCC 90028). However, the antimicrobial activity of (PJ) four fractions used in this study was estimated using the broth microdilution method (7, 8).
Each PJ fraction was dissolved in 100% DMSO (dimethyl sulfoxide) (Riedeldehan, Germany) at a concentration of 100 mg/ml for hexane, methanol, and water fractions and 50 mg/ml for acetone fraction. The produced solution was filter-sterilized and then was serially microdiluted (2 folds) 11 times in sterile nutrient broth (Himedia, India). The dilution processes were performed under aseptic conditions in 96-well plates (Greiner bio-one, North America). In the microwells that were assigned to evaluate the antibacterial activities of the PJ leaf fractions, microwell number 11 contained plant free nutrient broth, which was used as a positive control for microbial growth. On the other hand, microwell number 12 contained plant-free nutrient broth that was left uninoculated with any of the test microbes. This well was used as a negative control for microbial growth. Microwell numbers 1–11 were inoculated aseptically with the test microbes. Each plant fraction was made in duplicate. All the inoculated plates were incubated at 35°C. The incubation period lasted for about 18 h for those plates inoculated with the test bacterial strains and for about 48 h for those plates inoculated with Candida albicans. The lowest concentration of PJ at which no visible microbial growth in that microwell was observed and considered the minimal inhibitory concentration (MIC) of the examined PJ plant four fractions (8).
2.8. Cell Culture and Cytotoxicity Assay
HeLa cervical adenocarcinoma cells were cultured in RPMI-1640 media, which was supplemented with 10% fetal bovine serum, 1% Penicillin/Streptomycin antibiotics, and 1% l-glutamine. Cells were grown in a humidified atmosphere with 5% CO2 at 37°C. Cells were seeded at cells/well in a 96-well plate. After 48 h, cells were incubated with various concentrations of the tested compounds for 24 h. Cell viability was assessed by CellTilter 96® Aqueous One Solution Cell Proliferation (MTS) Assay according to the manufacturer’s instructions (Promega Corporation, Madison, WI). Briefly, at the end of the treatment, 20 μl of MTS solution per 100 μl of media was added to each well and incubated at 37°C for 2 h. Absorbance was measured at 490 nm.
2.9. Statistical Analysis
The conducted tests were determined in triplicate for the four fractions of the PJ plant. The results were expressed as the (SD). Data were compared using unpaired -tests. The statistical significance was considered when the value was <0.05. Statistical significance is expressed in terms of when the value < 0.05, when the value ≤ 0.001, and when the value ≤ 0.0001.
3. Results
3.1. Phytochemical Screening
The conducted phytochemical analysis revealed the presence of tannins, saponins, and carbohydrates, in the PJ, while the flavonoids, phenols, amino acids, and cardiac glycosides were absent (Table 1).
Aqueous fraction
Hexane fraction
Acetone fraction
Methanol fraction
Phytochemical classes
—
—
—
—
Amino acids and protein
+++
—
—
+
Carbohydrate
—
—
—
++
Tannin
—
—
—
—
Flavonoid
—
—
—
—
Phenol
++
—
—
—
Saponin
—
—
—
—
Cardiac glycoside
—: no content; +: content; ++: high content.
... For thousands of years, people tried to cure their diseases by utilizing various available natural materials and among these were medicinal plants [1,2]. Recently, it is well recognized that diabetes mellitus is considered one of the most common diseases worldwide, which is characterized as a type of metabolic disorder known by hyperglycemia, hypoinsulinemia, hyperlipidemia, and increased oxidative stress [3]. ...
Teucrium leucocladum is among the most used traditional medicinal plants in Palestine, which is used for the treatment of hyperglycemia and colon spasms from ancient times. Therefore, the current investigation aimed for the first time to determine the hypoglycemic, hypolipidemic, and oxidative stress inhibitory effects of the aerial parts (stem and leaves) of T. leucocladum hydrophilic (water) extract in streptozotocin- (STZ-) induced diabetic rats (65 mg/kg), given intraperitoneally at a dose of 100 mg/kg for 21 days. The rats were divided into four groups as control (C), control + T. leucocladum extract (C + TL), diabetes (D), and diabetes + T. leucocladum extract (D + TL). The antioxidant activity was analyzed using in vitro 2,2-diphenyl-1-picrylhydrazyl and in vivo methods by measuring the plasma and tissue malondialdehyde (MDA) levels using a colorimetric assay. On the other hand, glutathione peroxidase (GSH-Px), erythrocyte superoxide dismutase (SOD) enzyme levels, serum paraoxonase (PON), and arylesterase (ARE) enzyme activities were assessed by utilizing standard biochemical kits. Besides, the blood glucose and serum insulin levels were assessed by a glucometer and Rat ELISA Kit, respectively. However, the autoanalyzer was used to evaluate the lipid profile. The diabetic rat group that administered T. leucocladum extract showed the best reduction in the tissue and plasma MDA levels and an increase of insulin-releasing potentials. Besides, the serum PON and ARE activities and erythrocyte superoxide dismutase and whole blood glutathione peroxidase enzyme levels were significantly increased in all animals treated with T. leucocladum extract. The current investigation demonstrated that T. leucocladum manifests antihyperglycemic and antihyperlipidemic effects and also increased the antioxidative defense system and reduced the lipid peroxidation process in experimental diabetic rats.
... Finally, 20 μL of MTS solution per 100 μL of media was added to each well and the plate was incubated at 37 • C for 2 h. The absorbance was recorded at 490 nm (Amer et al., 2019). ...
The essential oils extracted from plants play an important role in medicine in addition to their huge value in the cosmetics and nutraceutical industries. In this investigation, besides the identification and quantification of the phytochemical composition of Nepeta curviflora Boiss essential oil, in-vitro antioxidant, porcine pancreatic lipase, α-glucosidase, and α-amylase inhibitory activities of the essential oil obtained by microwave ultrasonic-assisted extraction were investigated. More importantly, it was designed to assess the essential oil impact on cell migration and cell proliferation of cervical cancer cell lines. The phytochemical composition of Nepeta curviflora essential oil was qualitatively and quantitatively determined utilizing gas chromatography-mass spectrometry technique. Twenty compounds were identified from Nepeta curviflora essential oil representing 100% of total essential oil, of which 1,6-dimethyl spiro[4.5]decane (27.51%), caryophyllene oxide (20.08%), and β-caryophyllene (18.28%) were the most abundant compounds. The antioxidant activity of the essential oil was measured as 6.3 ± 0.43 μg/mL in comparison with trolox (positive control) employing the 1,1-diphenyl-2-picrylhydrazyl radical scavenging assay. The high concentrations of the caryophyllene oxide and caryophyllene thought to contribute significantly to the antioxidant of the essential oil. The obtained essential oil displayed α-glucosidase, α-amylase, and porcine pancreatic lipase inhibitory activities with half-maximal inhibitory concentration (IC50) values of 26.3 ± 0.57, 45.7 ± 0.26, and 54.9 ± 0.34 μg/mL, respectively, in comparison with the positive controls orlistat and acarbose which showed IC50 values of 12.3 ± 0.33, 28.84 ± 1.22 and 37.15 ± 0.32 μg/mL, respectively. Moreover, anticancer activity was established exploiting wound healing and cell proliferation assays against cervical cancer cell lines. Remarkably, non-cytotoxic concentrations of Nepeta curviflora essential oil have significantly inhibited the migration potential of cervical cancer cells after 24 h of treatment and revealed that the oil concentration and treatment time increase the inhibitory action on cervical cancer cell viability.
... Cell death (apoptosis and/or necrosis) was evaluated using Annexin V-FITC detection Kit as described in previous work [31]. After platting, U266 cells, they were incubated with extracts at 100 μg/mL or solvent control (methanol 0.5%) or H 2 O 2 (500 μM) for 24 h. ...
... In the human body, free radicals created by various biochemical processes such as the reduction of molecular oxygen in the aerobic respiration creating hydroxyl and superoxide radicals can be externally formed by splitting water molecules to produce the hydroxyl radicals using the gamma rays of electromagnetic radiation [1,2]. In fact, many studies demonstrated that the presence of uncontrolled free radicals in human body is usually correlated with broad diversity of illnesses such as asthma, atherosclerosis, diabetes, cancer, Alzheimer's disease, senile dementia, Parkinson's disease and obesity [3,4]. ...
Introduction
Throughout history, therapeutically active plant products have received substantial attention due to their valuable role in the discoveries of specific medications. The aim of this study was to assess, for the first time, the antimicrobial, antioxidant, antilipase, anti-α-amylase and cytotoxic properties of four fractions derived from Anchusa ovata Lehm. (AO) leaves.
Methods
Antioxidant, antilipase and anti-amylase potentials of (AO) were established using DPPH (1,1-diphenyl-2-picrylhydrazyl), p-nitrophenyl butyrate and dinitro-salicylic acid procedures, respectively, while antimicrobial activity was conducted using broth microdilution assay against eight Gram-positive, Gram-negative bacterial strains in addition to one fungal strain. Moreover, the MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] cytotoxic assay was utilized against cervical cancer cells (HeLa).
Results
The methanol fraction of AO showed potential antioxidant, antilipase, and α-amylase inhibitory activities with IC50 values of 9.55 ± 0.13, 53.7 ± 0.41 and 16.55 ± 1.84 μg/ml, respectively compared with the positive controls Trolox, Orlistat and Acarbose that had IC50 values of 3.23 ± 0.92, 12.3 ± 0.35 and 28.18 ± 1.22 μg/ml, respectively. Moreover, the hexane, acetone, and methanol fractions had wide ranges of antimicrobial potential. In addition, the cytotoxic activity outcomes which showed the best activity was for the aqueous followed by acetone, hexane and methanol fractions with IC50 values of 1.04, 2.72, 3.96 and 17.67 mg/ml, respectively.
Conclusion
Our data demonstrate a wide range of biological characteristics for each AO plant fraction. This profiling information about the methanol fraction provided important data for further research and pharmaceutical applications.
... In the human body, free radicals created by various biochemical processes such as the reduction of molecular oxygen in the aerobic respiration creating hydroxyl and superoxide radicals can be externally formed by splitting water molecules to produce the hydroxyl radicals using the gamma rays of electromagnetic radiation [1,2]. In fact, many studies demonstrated that the presence of uncontrolled free radicals in human body is usually correlated with broad diversity of illnesses such as asthma, atherosclerosis, diabetes, cancer, Alzheimer's disease, senile dementia, Parkinson's disease and obesity [3,4]. ...
Aim. To study the anatomical structure of Cercis siliquastrum L. leaves. Materials and methods. The study objects were leaves of Judas tree collected in the Botanical garden of the National University of Pharmacy in June, 2021. Micropreparations were made of fresh, dried material and fixed with a mixture of 96 % ethanol, water and glycerin (1 : 1: 1). Micropreparations were prepared and examined according to the methodsof the SPhU using a Granum microscope (Austria). A Canon PC1251 camera was used for photographing. Results and discussion. The type of the leaf blade is dorsoventral, hypostomatic, there are cells with the orange content. There are numerous stomata of anocytic (sometimes anisocytic) type. Simple multicellular hairs with a weakly warty cuticle were found on the lower and upper epidermis. A 2-3-layered collenchymatous parenchyma is located above the central vein. The bundle sheath is crystalliferous. On the xylem side, 5-6 layers of the sclerenchyma are observed. The petiole has an oval shape, slightly ribbed near the leaf blade. In the middle of the petiole there are three bundles: two radial on the sides and one central with two additional bundles. All bundles are surrounded by a common sclerenchymal ring. There are cells with the orange content in the parenchyma. On the abaxial side there is a layer of cells with calcium oxalate drusens under the sclerenchyma. The petiole is rounded, slightly ribbed. The bundle has a crescent shape in the center. The xylem rays are with the brown content. The sclerenchyma is well developed. There are drusens in the parenchyma of the petiole. Conclusions. The main anatomical features of Judas tree leaves have been determined; it will allow identifying and standardizing the plant raw material. The data obtained are necessary for the development of Drug Quality Control Methods (DQCM) for the medicinal raw material.
Introduction
Tamarindus indica (T. indica) is a tropical tree with multiple purposes and is used in many diets and traditional remedies. Apart from the antioxidant capacity and polyphenolic content, the seeds of T. indica have also shown anti-proliferative activities but no studies have been reported on colorectal cancer. Hence, this study aims to explore the anti-proliferative effects of the methanol extract of T. indica seeds (TSM) in colorectal cancer.
Methods
The colorectal cancer cell line (HT-29) was treated with TSM. Mechanisms for the anti-proliferative effects of TSM were determined by measuring parameters of oxidative stress, caspases, NF-κB activity and cell cycle. Apoptosis in the treated cells was confirmed by measuring expression of apoptotic proteins using the human apoptosis antibody array kit. Polyphenolic compounds in HT-29 cells treated with TSM were identified using UHPLC.
Results
TSM inhibited the growth of HT-29 cells (IC50 = 75.76 ± 0.83 µg/ml), and showed low cytotoxicity towards normal colon cells (CCD 841 CoN). The treatment caused alteration in redox status, oxidative damage, activation of caspases-3/7, -8 and -9, reduction in NF-κB activity, and cell cycle arrest. Activation of apoptosis was confirmed by protein array data, showing upregulation of pro-apoptotic proteins. UHPLC analyses identified the presence of three polyphenolic compounds (gallic acid, ferulic acid, and naringenin) in TSM-treated HT-29 cells.
Conclusion
The apoptotic effect of TSM was possibly mediated by reactive oxygen and nitrogen species (ROS/RNS) insult, affecting various signalling pathways, eventually causing apoptosis of HT-29 cells. This extract has great potential as a promising natural adjunct therapy for colorectal cancer.
This research involved antioxidant screening and evaluation of total phenol and flavonoid contents of ethanolic extracts from 30 Iranian plant species. Total phenol content was determined for each extract using the Folin-Ciocalteu method and total flavonoids was assessed by the Dowd method. A high phenol content was detected for Loranthus grewinkii (35.32±0.31 mg gallic acid equivalent (GAE)/g) of dry plant followed by Pteropyrum olivieri, Phoenix dactylifera, Cercis griffithii and Lippia citriodora. While relatively low levels of flavonoid content were detected for tested plants except in Pteropyrum olivieri) 14.53±0.13mg QE/g of dry plant). Free radical scavenging activity was determined using DPPH (1,1-diphenyl-2-picrylhydrazyl), NO (Nitric Oxide) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)free radical assays. Ferric Reducing Antioxidant Power (FRAP) assay was conducted as a measure of antioxidant capacity. L. grewinkii was superior in DPPH, NO and ABTS free radical inhibition. Extract of P. olivieri demonstrated a potent inhibitory activity against NO free radical compared to quercetin. Based on overall antioxidant activity, L. grewinkii was determined as the strongest in terms of free radical scavenging effect. A positive correlation observed between phenolic content and the activity while the flavonoids may have major contribution to manifestation of antioxidant activity in most of the investigated plant species.
Ovarian cancer is a heterogeneous disease that encompasses a number of different cellular subtypes, the most common of which is high-grade serous ovarian cancer (HGSOC). Still today, ovarian cancer is primarily treated with chemotherapy and surgery. Recent advances in the hereditary understanding of this disease have shown a significant role for the BRCA gene. While only a minority of patients with HGSOC will have a germline BRCA mutation, many others may have tumor genetic aberrations within BRCA or other homologous recombination proteins. Genetic screening for these BRCA mutations has allowed improved preventative measures and therapeutic development. This review focuses on the understanding of BRCA mutations and their relationship with ovarian cancer development, as well as future therapeutic targets.
Experimental studies have shown that a variety of chemopreventive plant components affect tumor initiation, promotion, and progression and the main difference, between botanical medicines and synthetic drugs, resides in the presence of complex metabolite mixtures shown by botanical medicine which in turn exert their action on different levels and via different mechanisms. In the present study, we performed an in vitro screening of ethanol extracts from commercial plants in order to investigate potential antitumor activity against human tumor cell lines. Experimental results obtained through a variety of methods and techniques indicated that extracts of I. verum , G. glabra , R. Frangula , and L. usitatissimum present significant reduction in in vitro tumor cell proliferation, suggesting these extracts as possible chemotherapeutical adjuvants for different cancer treatments.
Herbal therapy is a holistic therapy, integrating emotional, mental and spiritual levels. Life style, emotional, mental and spiritual considerations are part of any naturopathic approach. The use of herbs does not generally involve "drug" actions or adverse effects. Although medicinal plants are widely used and assumed to be safe, however, they can potentially be toxic. Where poisoning from medicinal plants has been reported, it usually has been due to misidentification of the plants in the form, in which they are sold, or incorrectly preparation and administration by inadequately trained personnel. There are some "drug like" plants remedies that their actions approach that of pharmaceuticals. Herbalists use these plants in treatment strategies and in countries such as Britain their vast availability is restricted by law. Digitalis is one of these examples and the number of these plants is not a lot. The mechanisms by which the herbs generally act are not established, however, most of medicinal plants possess antioxidant activities. The plants have been shown to effective by this property is various conditions including cancer, memory deficit and Alzheimer, atherosclerosis, diabetes and other cardiovascular diseases. Antioxidant activities of herbal medicines are also effective in reducing the toxicities of toxic agents or other drugs.
1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) is the first committed enzyme in the 2-methyl-D-erythritol 4-phosphate (MEP) terpenoid biosynthetic pathway and is also a validated antimicrobial target. Theaflavins, which are polyphenolic compounds isolated from fermented tea, possess a wide range of pharmacological activities, especially an antibacterial effect, but little has been reported on their modes of antimicrobial action. To uncover the antibacterial mechanism of theaflavins and to seek new DXR inhibitors from natural sources, the DXR inhibitory activity of theaflavins were investigated in this study. The results show that all four theaflavin compounds could specifically suppress the activity of DXR, with theaflavin displaying the lowest effect against DXR (IC50 162.1 μM) and theaflavin-3,3′-digallate exhibiting the highest (IC50 14.9 μM). Moreover, determination of inhibition kinetics of the theaflavins demonstrates that they are non-competitive inhibitors of DXR against 1-deoxy-D-xylulose 5-phosphate (DXP) and un-competitive inhibitors with respect to NADPH. The possible interactions between DXR and the theaflavins were simulated via docking experiments.
Background:
The burden of cancer is difficult to study in the context of the occupied Palestinian territory because of the limited data available. This study aims to evaluate the quality of mortality data and to investigate cancer mortality patterns in the occupied Palestinian territory's West Bank governorates from 1999 to 2009.
Methods:
Death certificates collected by the Palestinian Ministry of Health for Palestinians living in the West Bank were used. Direct and indirect age-standardised mortality rates were computed and used to compare different governorates according to total and specific cancer mortality. Furthermore, standardised proportional mortality ratios were calculated to compare mortality by urban, rural and camp locales.
Results:
The most common cause of death out of all cancer types was lung cancer among males (22.8 %) and breast cancer among females (21.5 %) followed by prostate cancer for males (9.5 %) and by colon cancer for females (11.4 %). Regional variations in cancer-specific causes of death were observed. The central- West Bank governorates had the lowest mortality for most cancer types among men and women. Mortality for lung cancer was highest in the north among men (SMR 109.6; 95%CI 99.5-120.4). For prostate cancer, mortality was highest in the north (SMR 103.6; 95%CI 88.5-120.5) and in the south (SMR 118.6; 95%CI 98.9-141.0). Breast cancer mortality was highest in the south (SMR 119.3; 95%CI 103.9-136.2). Similar mortality rate patterns were found in urban, rural and camp locales.
Conclusion:
The quality of the Palestinian mortality registry has improved over time. Results in the West Bank governorates present different mortality patterns. The differences might be explained by personal, contextual and environmental factors that need future in-depth investigations.
DNA is vulnerable to damage resulting from endogenous metabolites, environmental and dietary carcinogens, some anti-inflammatory drugs, and genotoxic cancer therapeutics. Cells respond to DNA damage by activating complex signalling networks that decide cell fate, promoting not only DNA repair and survival but also cell death. The decision between cell survival and death following DNA damage rests on factors that are involved in DNA damage recognition, and DNA repair and damage tolerance, as well as on factors involved in the activation of apoptosis, necrosis, autophagy and senescence. The pathways that dictate cell fate are entwined and have key roles in cancer initiation and progression. Furthermore, they determine the outcome of cancer therapy with genotoxic drugs. Understanding the molecular basis of these pathways is important not only for gaining insight into carcinogenesis, but also in promoting successful cancer therapy. In this Review, we describe key decision-making nodes in the complex interplay between cell survival and death following DNA damage.
Medicinal plants have historically proven their value as a source of molecules with therapeutic potential, and nowadays still represent an important pool for the identification of novel drug leads. In the past decades, pharmaceutical industry focused mainly on libraries of synthetic compounds as drug discovery source. They are comparably easy to produce and resupply, and demonstrate good compatibility with established high throughput screening (HTS) platforms. However, at the same time there has been a declining trend in the number of new drugs reaching the market, raising renewed scientific interest in drug discovery from natural sources, despite of its known challenges. In this survey, a brief outline of historical development is provided together with a comprehensive overview of used approaches and recent developments relevant to plant-derived natural product drug discovery. Associated challenges and major strengths of natural product-based drug discovery are critically discussed. A snapshot of the advanced plant-derived natural products that are currently in actively recruiting clinical trials is also presented. Importantly, the transition of a natural compound from a "screening hit" through a "drug lead" to a "marketed drug" is associated with increasingly challenging demands for compound amount, which often cannot be met by re-isolation from the respective plant sources. In this regard, existing alternatives for resupply are also discussed, including different biotechnology approaches and total organic synthesis. While the intrinsic complexity of natural product-based drug discovery necessitates highly integrated interdisciplinary approaches, the reviewed scientific developments, recent technological advances, and research trends clearly indicate that natural products will be among the most important sources of new drugs also in the future.
Copyright © 2015. Published by Elsevier Inc.
Medicinal plants are being used as desirable natural sources of various preparations made in traditional herbal therapy worldwide. Since ancient times, traditional remedies have been trusted by Iranian people. Plants of various families can be found in the prescription of rural healers in all regions of Iran. Some of these are native to Iran but many others are known as exotic plants which only grow in especial regions worldwide. This study for the first time illustrates the presence and the growth of several families of these plants in the climate of Iran along with some of their most important pharmacological effects and their active constituents.
With tens of thousands of plant species on earth, we are endowed with an enormous wealth of medicinal remedies from Mother Nature. Natural products and their derivatives represent more than 50% of all the drugs in modern therapeutics. Because of the low success rate and huge capital investment need, the research and development of conventional drugs are very costly and difficult. Over the past few decades, researchers have focused on drug discovery from herbal medicines or botanical sources, an important group of complementary and alternative medicine (CAM) therapy. With a long history of herbal usage for the clinical management of a variety of diseases in indigenous cultures, the success rate of developing a new drug from herbal medicinal preparations should, in theory, be higher than that from chemical synthesis. While the endeavor for drug discovery from herbal medicines is "experience driven," the search for a therapeutically useful synthetic drug, like "looking for a needle in a haystack," is a daunting task. In this paper, we first illustrated various approaches of drug discovery from herbal medicines. Typical examples of successful drug discovery from botanical sources were given. In addition, problems in drug discovery from herbal medicines were described and possible solutions were proposed. The prospect of drug discovery from herbal medicines in the postgenomic era was made with the provision of future directions in this area of drug development.
In recent years, there has been a great deal of attention toward the field of free radical chemistry. Free radicals reactive oxygen species and reactive nitrogen species are generated by our body by various endogenous systems, exposure to different physiochemical conditions or pathological states. A balance between free radicals and antioxidants is necessary for proper physiological function. If free radicals overwhelm the body's ability to regulate them, a condition known as oxidative stress ensues. Free radicals thus adversely alter lipids, proteins, and DNA and trigger a number of human diseases. Hence application of external source of antioxidants can assist in coping this oxidative stress. Synthetic antioxidants such as butylated hydroxytoluene and butylated hydroxyanisole have recently been reported to be dangerous for human health. Thus, the search for effective, nontoxic natural compounds with antioxidative activity has been intensified in recent years. The present review provides a brief overview on oxidative stress mediated cellular damages and role of dietary antioxidants as functional foods in the management of human diseases.
Abstract The current understanding of the complex role of ROS in the organism and pathological sequelae of oxidative stress points to the necessity of comprehensive studies of antioxidant reactivities and interactions with cellular constituents. Studies of antioxidants performed within the COST B-35 action has concerned the search for new natural antioxidants, synthesis of new antioxidant compounds and evaluation and elucidation of mechanisms of action of both natural and synthetic antioxidants. Representative studies presented in the review concern antioxidant properties of various kinds of tea, the search for new antioxidants of herbal origin, modification of tocopherols and their use in combination with selenium and properties of two promising groups of synthetic antioxidants: derivatives of stobadine and derivatives of 1,4-dihydropyridine.
Introduction
Protection of skin from wrinkles and pigmentation changes are among the most common challenges for modern science and cosmetics. These are caused by unusual disruption of connective tissue, the formation of free radicals and ultraviolet radiation.
The current research is the first study to compare the phytoconstituents, antioxidant, sun protection factor (SPF) and skin wrinkles prevention effects using four different solvents fractions for the traditional plant Alkanna tinctoria (L.) root bark.
Methods
Qualitative and quantitative phytoconstituents, antioxidant activity, sun protection factor and anti-elastase (skin wrinkles prevention) effects were assessed using standard pharmaceutical and cosmeceuticals assays.
Results
The results of antioxidant, anti-elastase and sun protection factor activities were the highest for the acetone fraction with IC50 8.51 ± 1.94 µg/ml, 10.02 ± 0.3 µg/ml and SPF value of 6.38, respectively. However, these results may occur due to its high contents of flavonoids and phenols which were 26.55 ± 1.6 mg of QUE/g and 59.48 ± 0.56 mg of GAE/g plant acetone fraction, respectively.
Conclusion
The acetone fraction of A. tinctoria root bark could be a promising candidate for cosmetic and pharmaceutical preparations due to the potential antioxidant, anti-elastase enzyme and sun protection activities.
Association of oxidative stress with carcinogenesis is well known, but not understood well, as is pathophysiology of oxidative stress generated during different types of anti-cancer treatments. Moreover, recent findings indicate that cancer associated lipid peroxidation might eventually help defending adjacent nonmalignant cells from cancer invasion. Therefore, untargeted metabolomics studies designed for advanced translational and clinical studies are needed to understand the existing paradoxes in oncology, including those related to controversial usage of antioxidants aiming to prevent or treat cancer. In this short review we have tried to put emphasis on the importance of pathophysiology of oxidative stress and lipid peroxidation in cancer development in relation to metabolic adaptation of particular types of cancer allowing us to conclude that adaptation to oxidative stress is one of the main driving forces of cancer pathophysiology. With the help of metabolomics many novel findings are being achieved thus encouraging further scientific breakthroughs. Combined with targeted qualitative and quantitative methods, especially immunochemistry, further research might reveal bio-signatures of individual patients and respective malignant diseases, leading to individualized treatment approach, according to the concepts of modern integrative medicine.
Natural products (NPs) are secondary metabolites produced and used by organisms for defending or adapting purposes. These molecules were naturally selected during thousands of years to improve the specificity and cover a very wide range of functions, depending on the origin, the habitat and the specific activity carried out in the organism of origin. Due to these intrinsic features, NPs have been used as healing agents since thousands of years and still today continue to be the most important source of new potential therapeutic preparations.The purpose of this review is to provide information about the historical evolution of the NPs investigation methods, focusing attention on the relative benefit/problems emerged after the improvement of the scientific investigations about them, especially over the last two centuries. Taken together, the reported information lead to the central role of NPs in the future of drug development for human needs.
Purpose
To determine if the magnetic resonance imaging (MRI) of the head and neck can predict distant metastases (DM) from nasopharyngeal carcinoma (NPC).
Methods and materials
MRI examinations of 763 NPC patients were assessed for primary tumour stage (T), nodal stage (N), primary tumour volume (PTV) and total nodal volume (NV). The association between MRI and clinical parameters were examined in DM+ and DM− patients using logistic regression and for distant metastases free survival (DMFS) using cox regression. Optimum thresholds were assessed by receiver-operating characteristics analysis, and positive predictive value (PPV) and odds ratio (OR) calculated.
Results
Distant metastases were present in 181/763 NPC patients (23.7%). Higher N stage and NV were the independent predictors of DM (p < 0.001 and 0.018 respectively) and poor DMFS (p = 0.001 and 0.030 respectively). Addition of NV (threshold ≥ 32.8 cm³) to the N stage improved the PPVs and ORs for DM in stage N1 (from 18.9% to 31.8% and 5.613 to 11.133 respectively) and stage N2 (from 40.4% to 60.8% and 16.189 to 36.979 respectively) but not in stage N3 (68.3% to 68.6% and 51.385 to 52.052 respectively).
Conclusion
MRI N stage and NV were independent predictors of DM and DMFS. The addition of NV in NPC patients with bulky N1 and N2 disease improved the ability of MRI to predict DM.
Decades after the first patients were treated with antibiotics, bacterial infections have again become a threat because of the rapid emergence of resistant bacteria-a crisis attributed to abuse of these medications and a lack of new drug development.
Traditional remedies are part of the cultural and religious life in Palestine. In
this survey, herbal products used in folk medicine were investigated and recorded.
A total of two hundred and sixteen (216) different plant species, sixty (60)
inorganic resources, and fifty two (52) animal products were in use. Among these
medicinal plants, twenty one (21) were used for their hypoglycemic effects,
eleven (11) were used for their hypotensive effects, and forty (40) were used for
their diuretic effects. Reservation of endangered medicinal plant species as well
as clinical investigation of these plants is required.
Abstract A total of 39 ethanol extracts of Jordanian plants were examined for cytotoxicity and mutagenicity. Among the tested plants, 8 extracts were active in the brine shrimp cytotoxicity bioassay with LC50 values in the range of 2.5-870 μg/ml. These positive extracts were further tested for their antitumor activity on human tumor cell lines in culture. Eminium spiculatum showed activity against a lung carcinoma line (A-549) with an ED50 of 0.0793 μg/ ml. Regarding mutagenic effect, 16 of the extracts were active on Salmonella typhim-urium strains TA98 or TA 102, and the extract of Podonosma oriantalis showed mutagenic activity in both strains.
Both carcinogenic and anticarcinogenic properties have been reported for the synthetic antioxidants butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). The association between dietary intake of BHA and BHT and stomach cancer risk was investigated in the Netherlands Cohort Study (NLCS) that started in 1986 among 120,852 men and women aged 55 to 69 years. A semi-quantitative food frequency questionnaire was used to assess food consumption. Information on BHA or BHT content of cooking fats, oils, mayonnaise and other creamy salad dressings and dried soups was obtained by chemical analysis, a Dutch database of food additives (ALBA) and the Dutch Compendium of Foods and Diet Products. After 6.3 years of follow-up, complete data on BHA and BHT intake of 192 incident stomach cancer cases and 2035 subcohort members were available for case-cohort analysis. Mean intake of BHA or BHT among subcohort members was 105 and 351 microg/day, respectively. For consumption of mayonnaise and other creamy salad dressings with BHA or BHT no association with stomach cancer risk was observed. A statistically non-significant decrease in stomach cancer risk was observed with increasing BHA and BHT intake [rate ratio (RR) highest/lowest intake of BHA = 0.57 (95% confidence interval (CI): 0.25-1.30] and BHT = 0.74 (95% CI: 0.38-1.43). In this study, no significant association with stomach cancer risk was found for usual intake of low levels of BHA and BHT.
Measurement of cellular DNA content and the analysis of the cell cycle can be performed by flow cytometry. Protocols for DNA measurement have been developed including Bivariate cytokeratin/DNA analysis, Bivariate BrdU/DNA analysis, and multiparameter flow cytometry measurement of cellular DNA content. This review summarises the methods for measurement of cellular DNA and analysis of the cell cycle and discusses the commercial software available for these purposes.
The three-dimensional culture of MCF-10A mammary epithelial cells on a reconstituted basement membrane results in formation of polarized, growth-arrested acini-like spheroids that recapitulate several aspects of glandular architecture in vivo. Oncogenes introduced into MCF-10A cells disrupt this morphogenetic process, and elicit distinct morphological phenotypes. Recent studies analyzing the mechanistic basis for phenotypic heterogeneity observed among different oncogenes (e.g., ErbB2, cyclin D1) have illustrated the utility of this three-dimensional culture system in modeling the biological activities of cancer genes, particularly with regard to their ability to disrupt epithelial architecture during the early aspects of carcinoma formation. Here we provide a collection of protocols to culture MCF-10A cells, to establish stable pools expressing a gene of interest via retroviral infection, as well as to grow and analyze MCF-10A cells in three-dimensional basement membrane culture.
Plant-derived compounds have been an important source of several clinically useful anti-cancer agents. These include vinblastine, vincristine, the camptothecin derivatives, topotecan and irinotecan, etoposide, derived from epipodophyllotoxin, and paclitaxel (taxol A number of promising new agents are in clinical development based on selective activity against cancer-related molecular targets, including flavopiridol and combretastin A4 phosphate, while some agents which failed in earlier clinical studies are stimulating renewed interest.
In Unani system of medicine, drugs consist of complex formulae with more than three components, for which, literature analysing these mixtures as they are sold in the market is scarce. In this paper, the main botanical components of the herbal tea known as "Zahraa" in Damascus, which contains between 6 and 14 species components is elucidated: Alcea damascena (Mout.) Mout. (Malvaceae), Aloysia triphylla (L'Herit.) Britt. (Malvaceae), Astragalus cf. amalecitanus Boiss., Cercis siliquastrum L. subsp. hebecarpa (Bornm.) Yalt. and subsp. siliquastrum. (Leguminosae), Colutea cilicica Boiss. et Bal. in Boiss. (Leguminosae), Crataegus aronia (L.) Bosc. ex DC. (Rosaceae), Cytisopsis pseudocytisus (Boiss.) Fertig. (Leguminosae), Eleagnus angustifolia L. (Eleagnaceae), Equisetum telmateia Ehrh. (Equisetaceae), Helichrysum stoechas (L.) Moench. subsp. barrelieri (Ten.) Nyman. (Compositae), Matricaria recutita L. (Compositae), Mentha longifolia L. subsp. noeana (Boiss. ex. Briq.) Briq. (Labiatae), Mentha spicata L. subsp. condensata (Briq.) Greuter and Burdet (Labiatae), Micromeria myrtifolia Boiss. and Hohen. in Boiss. (Labiatae), Paronychia argentea Lam. (Caryophyllaceae), Phlomis syriaca Boiss. (Labiatae), Rosa damascena Mill. (Rosaceae), Salvia fruticosa Mill. (Labiatae), Sambucus nigra L. (Caprifoliaceae), Spartium junceum L. (Leguminosae), Zea mays L. (Gramineae).
The recently discovered non-mevalonate pathway of isoprenoid biosynthesis serves as the unique source of terpenoids in numerous pathogenic eubacteria and in apicoplast-type protozoa, most notably Plasmodium, but is absent in mammalian cells. It is therefore an attractive target for anti-infective chemotherapy. The first committed step of the non-mevalonate pathway is catalyzed by 2C-methyl-D-erythritol 4-phosphate synthase (IspC). Using photometric and NMR spectroscopic assays, we screened extracts of Mediterranean plants for inhibitors of the enzyme. Strongest inhibitory activity was found in leaf extracts of Cercis siliquastrum.
Developing new antimicrobial therapies: are synergistic combinations of plant extracts/compounds with conventional antibiotics the solution?
- Cheesman