Search

The stability of cyclic peptides, coupled with their structural diversity and ability to host an extensive range of bioactivities, make them promising leads for the development of new drugs. PawS-Derived Peptide-23 (PDP-23) is a head-to-tail macrocyclic peptide with two disulfide bonds produced in plant seeds. Its unusual fold comprises two -hairpins connected by hinges that allow the structure to adapt to different environments. In water two PDP-23 molecules form a compact intertwined dimer that buries hydrophobic residues, whereas in membrane mimicking conditions it adopts an open monomeric form that expose them. Here we investigate PDP-23 as a novel scaffold for the grafting of bioactive epitopes and conjugation of small molecules. To explore the plasticity of PDP-23 we introduced the bioactive loop of sunflower trypsin inhbitor-1 (SFTI-1) or an integrin binding RGD motif into either of the -hairpins. Solution NMR spectroscopy revealed that although the variants were unable to dimerise, the structural features of both the graft and scaffold were retained. SFTI-1 hybrid variants showed trypsin inhibitory activity. PDP-23 has previously been used as a cell permeable drug scaffold targeting drug-resistant cancer cells by inhibiting the drug efflux pump P-glycoprotein and restoring their sensitivity to chemotherapeutic. Introducing the RGD motif into such PDP-23 conjugates significantly improve their potency, suggesting that the RGD sequence targets the peptide to the membrane of cancer cells and improve cell uptake. In conclusion, this study highlights PDP-23 as a stable and versatile scaffold for molecular grafting of bioactivities and targeted delivery of pharmaceutical payloads.

Head-to-tail cyclized peptides are intriguing natural products with unusual properties. The PawS-Derived Peptides (PDPs) are ribosomally synthesized as part of precursors for seed storage albumins in species of the daisy family, and are post-translationally excised and cyclized during proteolytic processing. Here we report a PDP twice the typical size and with two disulfide bonds, identified from seeds of Zinnia elegans. In water, synthetic PDP-23 forms a unique dimeric structure in which two monomers containing two β-hairpins cross-clasp and enclose a hydrophobic core, creating a square prism. This dimer can be split by addition of micelles or organic solvent and in monomeric form PDP-23 adopts open or closed V-shapes, exposing different levels of hydrophobicity dependent on conditions. This chameleonic character is unusual for disulfide-rich peptides and engenders PDP-23 with potential for cell delivery and accessing novel targets. We demonstrate this by conjugating a rhodamine dye to PDP-23, creating a stable, cell-penetrating inhibitor of the P-glycoprotein drug efflux pump., The cyclic peptide PDP-23 adopts a different structure depending on conditions. In water it forms a dimer, but can unfold allowing its hydrophobic core to interact with membranes. PDP-23 shows promise as a cell penetrating scaffold for drug delivery.

Head-to-tail cyclic and disulfide-rich peptides are natural products with applications in drug design. Among these are the PawS-Derived Peptides (PDPs) produced in seeds of the daisy plant family. PDP-23 is a unique member of this class in that it is twice the typical size and adopts two β-hairpins separated by a hinge region. The β-hairpins, both stabilised by a single disulfide bond, fold together into a V-shaped tertiary structure creating a hydrophobic core. In water two PDP-23 molecules merge their hydrophobic cores to form a square prism quaternary structure. Here, we synthesised PDP-23 and its enantiomer comprising d-amino acids and achiral glycine, which allowed us to confirm these solution NMR structural data by racemic crystallography. Furthermore, we discovered the related PDP-24. NMR analysis showed that PDP-24 does not form a dimeric structure and it has poor water solubility, but in less polar solvents adopts near identical secondary and tertiary structure to PDP-23. The natural role of these peptides in plants remains enigmatic, as we did not observe any antimicrobial or insecticidal activity. However, the plasticity of these larger PDPs and their ability to change structure under different conditions make them appealing peptide drug scaffolds., Larger members of the PawS-Derived family of cyclic plant peptides form complex structures. The graphical abstract shows the racemic crystal structure of the homodimeric PDP-23 as well as the solution NMR structure of PDP-24.

Background Tea (Camellia sinensis) is one of the most consumed beverages in the world. White tea is an unfermented tea made from young tea leaves or unopened buds. Methods The in vitro antiproliferation assay and in vivo Chemopreventive effects of white tea extract (WTE) using azoxymethane (AOM) induced aberrant crypt foci (ACF) in rats was assessed. Twenty-four adult male rats were injected 15 mg/kg body weight AOM subcutaneously once a week for two weeks and then were divided randomly into four groups. Group I (Cancer control), Group II (treated whit 200 mg/kg white tea extract), Group III (treated whit 400 mg/kg white tea extract) and group IV (35 mg/kg body weight 5-fluorouracil as reference control). On the last day of the experiment, the animals were euthanized using an overdose of ketamine and xylazine and the colon tissue was collected. The colon tissues were evaluated grossly and histopathologically for aberrant crypt foci (ACF). Prior to the cancer studies, the acute toxicity test was performed to demonstrate the safety usage of the extract. Results The antiproliferative activity of the extract was observed in HT-29 and HCT 116 colon cancer cells. The ACF score was significantly reduced following the white tea extract treatment in a dose-dependent manner. Furthermore, the acute toxicity study indicated that there were no nephrotoxic and hepatotoxic effects or any serum biochemical changes in the rats that were orally administered with 2000 mg/kg body weight of the extract. Conclusions This study suggested the the potential of white tea as a chemopreventative agent through inhibition of Azoxymethane-induced Colonic Aberrant Crypt Foci Formation in the male Sprague-Dawley rats.

Background Brucea javanica (L.) Merr. is a plant from the genus Brucea, which is used in local traditional medicine to treat various diseases. Recent studies revealed an impressive anticancer efficiency of B. javanica extract in different types of cancer cells. Purpose In this study, we have investigated the cytotoxic effects of the B. javanica hexane, ethanolic extracts against colon cancer cells. HT29 colon cells were selected as an in vitro cancer model to evaluate the anticancer activity of B. javanica ethanolic extract (BJEE) and the possible mechanisms of action that induced apoptosis. Methods 3-(4,5-dimethylthiazol-2-yl)-2, 5,-diphenyltetrazolium bromide (MTT), lactate dehydrogenase, acridine orange/propidium iodide, and annexin-V-fluorescein isothiocyanate assays were performed to determine the antiproliferative and apoptosis validation of BJEE on cancer cells. Measurement of reactive oxygen species (ROS) production, caspase activities, nucleus factor-κB activity, and gene expression experiments was done to investigate the potential mechanisms of action in the apoptotic process. Results The results obtained from this study illustrated the significant antiproliferative effect of BJEE on colorectal cancer cells, with a concentration value that inhibits 50% of the cell growth of 25±3.1 µg/mL after 72 h of treatment. MTT assay demonstrated that the BJEE is selectively toxic to cancer cells, and BJEE induced cell apoptosis via activation of caspase-8 along with modulation of apoptosis-related proteins such as Fas, CD40, tumor necrosis factor-related apoptosis-inducing ligands, and tumor necrosis factor receptors, which confirmed the contribution of extrinsic pathway. Meanwhile, increased ROS production in treated cells subsequently activated caspase-9 production, which triggered the intrinsic pathways. In addition, overexpression of cytochrome-c, Bax, and Bad proteins along with suppression of Bcl-2 illustrated that mitochondrial-dependent pathway also contributed to BJEE-induced cell death. Consistent with the findings from this study, BJEE-induced cancer cell death proceeds via extrinsic and intrinsic mitochondrial-dependent and -independent events. Conclusion From the evidence obtained from this study, it is concluded that the BJEE is a promising natural extract to combat colorectal cancer cells (HT29 cells) via induction of apoptosis through activation of extrinsic and intrinsic pathways.

Background Colorectal cancer is the third most common form of cancer in both men and women around the world. The chemistry and biological study of heterocyclic compounds have been an interesting area for a long time in pharmaceutical and medicinal chemistry. Methods A new synthetic compound, 2-(1,1-dimethyl-1H-benzo[e]indol-2-yl)-3-((2-hydroxyphenyl)amino) acrylaldehyde, abbreviated as DBID, was prepared through the reaction of 2-(diformylmethylidene)-1,1- dimethylbenzo[e]indole with 2-aminophenol. The chemical structure of the synthesized compound was characterized by 1H NMR, 13C NMR and APT-NMR spectroscopy and confirmed by elemental analysis (CHN). The compound was screened for the antiproliferation effect against colorectal cancer cell line, HCT 116 and its possible mechanism of action was elucidated. To determine the IC50 value, the MTT assay was used and its apoptosisinducing effect was investigated. Results DBID inhibited the proliferation of HCT 116 cells with an IC50 of 9.32 µg/ml and significantly increased the levels of caspase -8, -9 and -3/7 in the treated cells compared to untreated cells. Apoptosis features in HCT 116 cell was detected in treated cells by using the AO/PI staining that confirmed that the cells had undergone remarkable morphological changes in apoptotic bodies. Furthermore, this changes in expression of caspase -8, -9 and -3 were confirmed by gene and protein quantification using RT-PCR and western blot analysis, respectively. Conclusion The current study showed that the DBID compound has demonstrated chemotherapeutic activity which was evidenced by significant increases in the expression and activation of caspase and exploit the apoptotic signaling pathways to trigger cancer cell death.

Brucea javanica (L.) Merr. is a well-known plant in Chinese System of Medicine. Its fruits and seeds have been reported to possess curative properties against various ailments. The chemical constituents and biological activity of this plant have been an interesting area in plant and chemistry medicine. The aim of this study is to evaluate the antiproliferative effects of the B. javanica extract against a colon cancer cell line and identification of the chemical components derived from the extract. An ethanolic extract from B. javanica fruits was prepared by cold maceration method, subjected to LC-MS profiling to elucidate the composition abbreviated as BJEE. The extract was screened for the cytotoxicity effects on HCT-116 colon cancer cells via MTT and LDH methods. Additionally, AO/PI staining verified apoptosis features in HCT-116 cells through microscopic analysis. ROS, caspase activity, and gene expression has been performed to identify its possible mechanism of actions which contribute to apoptosis. Output data from this study showed BJEE inhibited the cell proliferation of HCT-116 colon cancer cells at IC50 value of 8.9 ± 1.32 (μg mL−1) and significantly increased the levels of caspase-8, 9, and 3/7 in treated cells in comparison to untreated. The changes in expression of caspase genes and some apoptosis genes like Bax and Bcl-2 were confirmed using RT-PCR. Phytochemical analysis by LC-MS identified six major active compounds (bruceine D, isobrucein A, quassimarin, C16 sphinganine, phytosphingosine, and enigmol) in BJEE that may play a key role in cell apoptosis. The current study showed BJEE could be a promising agent for colorectal cancer therapy by significant increase in caspase activity level, and up-regulation of the specific apoptotic genes.

Colorectal cancer is the third most common form of cancer affecting both men and women around the world. The chemical and biological studies of heterocyclic compounds have been an interesting area in pharmaceutical and medicinal chemistry. A new synthetic compound namely 2-(1,1-dimethyl-1H-benzo[e]indol-2-yl)-3-((2-hydroxyphenyl)amino)acrylaldehyde, abbreviated as DBID was screened for the antiproliferation effects against the colorectal cancer cell line, HT-29 and its possible mechanism of action was elucidated. To determine the IC50 value, MTT assay was employed and further verified by the LDH release assay and apoptosis-inducing effect. DBID inhibited the proliferation of HT-29 cells and significantly increased the levels of caspase -8, -9 and -3/7 in the treated cells compared to untreated cells. Apoptosis features in HT-29 cells were detected in treated cells by using the AO/PI staining and flow cytometric analysis of Annexin V. The changes in expression of some apoptotic genes were confirmed by gene quantification using RT-PCR. The current study showed that the DBID compound exhibited chemotherapeutic activity, which was evident by significant increases in the expression and activation of caspase, up-regulation of the expression of specific apoptotic genes and exploitation of the apoptotic signaling pathways to trigger cancer cell death.

Background/objective : The purpose of the present study was to evaluate the effect of fenugreek seed extract in combination with swimming exercise compared to glibenclamide consumption on type 2 diabetic rats. Design : The acute toxicity test was carried out to choose the safe doses and identify the toxicity effects of the fenugreek seed extract. To investigate the hypoglycemic effect of the extract and its effect in combination with swimming training, 80 Wistar Kyoto male streptozotocin-induced diabetic rats were divided randomly into eight groups: diabetic control (C); fenugreek seed extract 0.8 g/kg (F1); fenugreek extract 1.6 g/kg (F2); swimming training (S); swimming training plus fenugreek extract 0.8 g/kg (SF1); swimming training plus fenugreek extract 1.6 g/kg (SF2); glibenclamide (G) and swimming training plus glibenclamide (SG). The rats were orally administrated with the treatments once a day with the respective treatment, and the training groups were subjected to swimming training every day for 60 min. Fasting blood samples were collected to measure fasting blood glucose, lipid profile, adiponectin, leptin, and insulin concentrations. Results : The results obtained from acute toxicity study showed no toxicity effect of fenugreek seed extract on the tested dose. Biochemical analysis showed significant improvements in all of the groups compared to the control group ( p

Tea (Camellia sinensis) is the most highly consumed beverage in the world next to water. The common way of preparation is steeping in hot water which is varying for different type of tea. We investigated the antioxidant properties of 6 type of tea leaves under different time and temperatures of extraction method used. In general, all samples tested in this study demonstrated high levels of antioxidant capacity and antioxidant activity. The results indicate that the antioxidants activity is significantly affected by time and temperature of steeping and the highest was depending on the variety. White state values, green and black teas showed different levels of antioxidants under different extraction conditions. Overall, the highest activity for white tea was in prolonged hot and in some assays prolonged hot and cold extracts, whereas for green tea the highest activity observed in prolonged cold steeping while, for black tea was in short hot water infusion. The results of this study showed the antioxidant capacity of white and green tea was greater than black tea.

Elham Rouhollahi,1 Soheil Zorofchian Moghadamtousi,2 Fatemeh Hajiaghaalipour,3 Maryam Zahedifard,2 Faezeh Tayeby,2 Khalijah Awang,4 Mahmood Ameen Abdulla,3 Zahurin Mohamed1 1Pharmacogenomics Laboratory, Department of Pharmacology, Faculty of Medicine, 2Institute of Biological Sciences, Faculty of Science, 3Department of Biomedical Science, Faculty of Medicine, 4Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia Purpose: Curcuma purpurascens BI. is a member of Zingiberaceae family. The purpose of this study is to investigate the wound healing properties of hexane extract of C. purpurascens rhizome (HECP) against excisional wound healing in rats.Materials and methods: Twenty four rats were randomly divided into 4 groups: A) negative control (blank placebo, acacia gum), B) low dose of HECP, C) high dose of HECP, and D) positive control, with 6 rats in each group. Full-thickness incisions (approximately 2.00 cm) were made on the neck area of each rat. Groups 1–4 were treated two-times a day for 20 days with blank placebo, HECP (100 mg/kg), HECP (200 mg/kg), and intrasite gel as a positive control, respectively. After 20 days, hematoxylin and eosin and Masson’s trichrome stainings were employed to investigate the histopathological alterations. Protein expressions of Bax and Hsp70 were examined in the wound tissues using immunohistochemistry analysis. In addition, levels of enzymatic antioxidants and malondialdehyde representing lipid peroxidation were measured in wound tissue homogenates.Results: Macroscopic evaluation of wounds showed conspicuous elevation in wound contraction after topical administration of HECP at both doses. Moreover, histopathological analysis revealed noteworthy reduction in the scar width correlated with the enhanced collagen content and fibroblast cells, accompanied by a reduction of inflammatory cells in the granulation tissues. At the molecular level, HECP facilitates wound-healing process by downregulating Bax and upregulating Hsp70 protein at the wound site. The formation of new blood vessel was observed in Masson’s trichrome staining of wounds treated with HECP (100 and 200 mg/kg). In addition, HECP administration caused a significant surge in enzymatic antioxidant activities and a decline in lipid peroxidation.Conclusion: These findings suggested that HECP accelerated wound-healing process in rats via antioxidant activity, angiogenesis effect and anti-inflammatory responses involving Hsp70/Bax. Keywords: Zingiberaceae, wound closure, immunohistochemistry, antioxidant enzyme activity, inflammatory cells

Diabetes mellitus (DM) is a metabolic diseases characterized by hyperglycemia due to insufficient or inefficient insulin secretory response. This chronic disease is a global problem and there is a need for greater emphasis on therapeutic strategies in the health system. Phytochemicals such as flavonoids have recently attracted attention as source materials for the development of new antidiabetic drugs or alternative therapy for the management of diabetes and its related complications. The antidiabetic potential of flavonoids are mainly through their modulatory effects on glucose transporter by enhancing GLUT-2 expression in pancreatic β cells and increasing expression and promoting translocation of GLUT-4 via PI3K/AKT, CAP/Cb1/TC10 and AMPK pathways. This review highlights the recent findings on beneficial effects of flavonoids in the management of diabetes with particular emphasis on the investigations that explore the role of these compounds in modulating glucose transporter proteins at cellular and molecular level.

In the present study, we examined the cytotoxic effects of Schiff base complex, [N-(3,5-dichloro-2-oxidobenzylidene)-4-chlorobenzyhydrazidato](o-methylbenzyl)aquatin(IV) chloride, and C1 on MDA-MB-231 cells and derived breast cancer stem cells from MDA-MB-231 cells. The acute toxicity experiment with compound C1 revealed no cytotoxic effects on rats. Fluorescent microscopic studies using Acridine Orange/Propidium Iodide (AO/PI) staining and flow cytometric analysis using an Annexin V probe confirmed the occurrence of apoptosis in C1-treated MDA-MB-231 cells. Compound C1 triggered intracellular reactive oxygen species (ROS) production and lactate dehydrogenase (LDH) releases in treated MDA-MB-231 cells. The Cellomics High Content Screening (HCS) analysis showed the induction of intrinsic pathways in treated MDA-MB-231 cells, and a luminescence assay revealed significant increases in caspase 9 and 3/7 activity. Furthermore, flow cytometric analysis showed that compound C1 induced G0/G1 arrest in treated MDA-MB-231 cells. Real time PCR and western blot analysis revealed the upregulation of the Bax protein and the downregulation of the Bcl-2 and HSP70 proteins. Additionally, this study revealed the suppressive effect of compound C1 against breast CSCs and its ability to inhibit the Wnt/β-catenin signaling pathways. Our results demonstrate the chemotherapeutic properties of compound C1 against breast cancer cells and derived breast cancer stem cells, suggesting that the anticancer capabilities of this compound should be clinically assessed.

Saffron is consumed as food and medicine to treat several illnesses. This study elucidates the saffron effectiveness on diabetic parameters in-vitro and combined with resistance exercise in-vivo. The antioxidant properties of saffron was examined. Insulin secretion and glucose uptake were examined by cultured RIN-5F and L6 myotubes cells. The expressions of GLUT2, GLUT4 and AMPKα were determined by Western blot. Diabetic and non-diabetic male rats were divided into: control, training, extract treatment, training + extract treatment and metformin. The exercise and 40 mg/kg/day saffron treatments were carried out for six weeks. The antioxidant capacity of saffron was higher compare to positive control (P 0.05). Serum glucose, cholesterol, triglyceride, low-density lipoprotein, very low-density lipoprotein, insulin resistance and glycated hemoglobin levels decreased in treated rats compared to untreated (p 0.05). The findings suggest that saffron consuming alongside exercise could improve diabetic parameters through redox-mediated mechanisms and GLUT4/AMPK pathway to entrap glucose uptake.

Injury to podocytes is an early event in diabetic nephropathy leading to proteinuria with possible progression to end-stage renal failure. The podocytes are unique and highly specialized cells that cover the outer layer of kidney ultra-filtration barrier and play an important role in glomerular function. In the past few decades, adult stem cells, such as mesenchymal stem cells (MSCs) with a regenerative and differentiative capacity have been extensively used in cell-based therapies. In addition to their capability for regeneration and differentiation, MSCs contributes to their milieu by paracrine action of a series of growth factors via antiapoptotic, mitogenic and other cytokine actions that actively participate in treatment of podocyte damage through prevention of podocyte effacement, detachment and apoptosis. It is hoped that novel stem cell-based therapies will be developed in the future to prevent podocyte injury, thereby reducing the burden of kidney disease.

Tea (Camellia sinensis) is the most highly consumed beverage in the world next to water. The common way of preparation is steeping in hot water which is varying for different type of tea. We investigated the antioxidant properties of 6 type of tea leaves under different time and temperatures of extraction method used. In general, all samples tested in this study demonstrated high levels of antioxidant capacity and antioxidant activity. The results indicate that the antioxidants activity is significantly affected by time and temperature of steeping and the highest was depending on the variety. White state values, green and black teas showed different levels of antioxidants under different extraction conditions. Overall, the highest activity for white tea was in prolonged hot and in some assays prolonged hot and cold extracts, whereas for green tea the highest activity observed in prolonged cold steeping while, for black tea was in short hot water infusion. The results of this study showed the antioxidant capacity of white and green tea was greater than black tea.

Camellia sinensis(tea) is reported to have health benefits, including the building of healthy skin. This study evaluated the effects of topical application ofCamellia sinensisextract on the rate of wound closure and the histology of wound area. A uniform area of 2.00 cm in diameter was excised from the neck of adult male Sprague Dawley rats. The animals were topically treated with 0.2 mL of vehicle (CMC), Intrasite gel (positive control), or 200 and 400 mg/mL of extract. Wounds dressed with the extract and Intrasite gel healed significantly earlier than those with vehicle. Histological analysis of the wound area after 10 days showed that wounds dressed with the extract had less scar width when compared to the control. The tissue contained less inflammatory cells and more collagen and angiogenesis, compared to wounds dressed with vehicle. In this study,Camellia sinensisshowed high potential in wound healing activity.

Tea (Camellia sinensis) is one of the most consumed beverages in the world. White tea is made from the buds and young leaves of the tea plant which are steamed and dried, whilst undergoing minimal oxidation. The MTT assay was used to test the extract on the effect of the proliferation of the colorectal cancer cell line, HT-29. The extract inhibited the proliferation of HT-29 cells with an IC50 of 87μg/ml. The extract increased the levels of caspase-3, -8, and -9 activity in the cells. DNA damage in 3T3-L1 normal cells was detected by using the comet assay. The extract protected 3T3-L1 cells against H2O2-induced DNA damage. The results from this study show that white tea has antioxidant and antiproliferative effects against cancer cells, but protect normal cells against DNA damage. Regular intake of white tea can help to maintain good health and protect the body against disease.