Objective: To evaluate the anti-obesity activity of ethanolic extract of cashew apple using various in vitro and in vivo models.
Methods: Phytochemical screening was carried out in ethanolic extract of cashew apple, followed by quantification of phenol and flavonoid. Antioxidant potential was evaluated using 2,2-diphenyl-1-picrylhydrazyl and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) scavenging assays. The inhibitory effect of ethanolic extract of cashew apple on α-amylase and pancreatic lipase was also studied. In addition, anti-obesity activity was determined in two in vivo models, lipid emulsion model and atherogenic diet-induced obese rat model. Levels of postprandial plasma triglycerides were assessed in lipid emulsion model, whereas serum lipid profile, in vivo antioxidants and histopathological studies of the carotid artery and liver were performed in an atherogenic diet-induced obese model.
Results: Phytochemical screening revealed the presence of carbohydrates, alkaloids, polyphenols, terpenoids, and steroids. The in vitro assays showed inhibition of α-amylase and pancreatic lipase and strong antioxidant potential. Ethanolic extract of cashew apple showed significant and time- dependent inhibitory activity on postprandial triglycerides after administration of lipid emulsion for 5 h. Ethanolic extract of cashew apple at 200 and 400 mg/kg on day 60 showed a significant reduction in body weight, body mass index and atherogenic index, whereas lipid profile and liver function marker levels in the serum were decreased in a dose-dependent manner at time intervals (day 0, 20, 40, and 60) compared to the atherogenic diet-induced obese rats. Histological observations showed reduced non-alcoholic fatty liver deposits and decreased atherosclerotic fatty streak plaques (carotid artery) after treatment with ethanolic extract of cashew apple.
Conclusions: Ethanolic extract of cashew apple ameliorates obesity, which may be partly mediated by its delayed absorption of cholesterol and carbohydrates.
Objective: To investigate the effect of alpha-lipoic acid (ALA) supplementation on systolic blood pressure (SBP), renal oxidant-antioxidant status and renal damage in spontaneously hypertensive rats (SHR) and SHR administered with Nω-nitro-L-arginine methyl ester (L-NAME).
Methods: Male rats were divided into four groups (SHR, SHR+ALA, SHR+L-NAME, SHR+ALA+L-NAME). The respective group of rats was administered with ALA (100 mg/ kg/day) from age 4 weeks to 28 weeks and L-NAME (25 mg/kg/day) from age 16 weeks to 28 weeks. SBP was measured every two weeks and twenty four hour urine was collected at 4 weeks, 16 weeks and 28 weeks for estimation of protein, creatinine and N-acetyl-β-D-glucosaminidase. At the end of 28 weeks, rats were sacrificed and blood and kidneys collected for assessment of blood creatinine, kidney thiobarbituric acid reactive substances, protein carbonyls, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase, glutathione disulfide, glutathione, total antioxidant status and nitric oxide as well as histopathological examination.
Results: ALA supplementation significantly reduced SBP of SHR and SHR+L-NAME rats when compared to their respective non-supplemented groups. Renal oxidant status markers including thiobarbituric acid reactive substances and protein carbonyls were significantly reduced on SHR and SHR+L-NAME rats supplemented with ALA at 28 weeks as well as ALA supplementation significantly increased renal antioxidants including superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione and glutathione/ glutathione disulfide ratio at 28 weeks. No significant change in nitric oxide levels was observed between the ALA supplemented and non-supplemented groups. Renal dysfunction was ameliorated on ALA supplementation as evidenced by significant reduction in urine protein levels, N-acetyl-β-D-glucosaminidase activity and significant increase of creatinine clearance in SHR and SHR+L-NAME at 28 weeks. Renal histopathological examination showed that ALA supplementation prevented vascular damage in SHR and ameliorated glomerular damage in SHR+L-NAME at 28 weeks.
Conclusions: ALA has hypotensive and renoprotective effects on both SHR and SHR+L-NAME, which could be due to its ability to ameliorate oxidative stress in the kidneys.
Objective: To evaluate the therapeutic potential of hydroalcoholic extract of licorice root against ethanol and cerulein induced chronic pancreatitis in rats.
Methods: The phytochemical profile of hydroalcoholic extract of licorice root was determined by high performance liquid chromatography (HPLC) and gas chromatography coupled to mass spectrometry (GC-MS). Chronic pancreatitis was induced in male albino Wistar rats by feeding them a diet containing ethanol (0%-36% of total calories) for 4 weeks and cerulein (20 μg/kg b.wt, i.p.) thrice a week for 3 weeks. Lipase and amylase in serum, lipid peroxides and antioxidants including reduced glutathione, glutathione peroxidase, superoxide dismutase and catalase in pancreas were determined. Inflammatory response was measured by myeloperoxidase in the pancreas, caspase-1 and the concentrations of IL-1 β and IL-18 in serum. Moreover, histological evaluation of the pancreas and liver was carried out.
Results: Different flavonoids and saponins were identified in the hydroalcoholic extract of licorice root through HPLC and GC-MS. A marked increase in the levels of serum lipase, amylase, lipid peroxides, caspase-1, myeloperoxidase, IL-1 β , and IL-18 and a marked decrease in the levels of antioxidants were observed after ethanol and cerulein administration. Treatment with hydroalcoholic extract of licorice root attenuated these changes. In addition, histological observation confirmed the protective effect of the extract in the pancreas and liver against inflammatory changes induced by ethanol and cerulein.
Conclusions: The licorice root extract attenuates ethanol and cerulein induced pancreatitis in rats probably due to its antioxidant phytonutrients since ethanol and cerulein-induced production of reactive oxygen species contributes to severe inflammation in the pancreas.
Objective: To identify and isolate phenolic compounds from Cuspidaria convoluta, and to evaluate their antibacterial activity and synergistic effect with antibiotics.
Methods: The crude extract was prepared by maceration with methanol (5%). The dry extract was suspended in water and fractionated successively. The most active extract was selected by its antibacterial activity and its total phenol content was determined by spectrophotometry and by HPLC-MS/MS. Bioactive fractions of the most active extract were separated by column chromatography and evaluated by bioautography. Isolated compounds were identified. Minimum inhibitory concentration (MIC) of these compounds was determined by microdilution broth method, and synergism with antibiotics (ampicillin, gentamicin and oxacillin) was tested by checkerboard and time-kill assays.
Results: Coumaric acid, catechin/epicatechin, and luteolin were purified and identified from the extract. There was an increase in the antibacterial activity of antibiotics when they were combined with these compounds. The combination of luteolin and ampicillin had the most potent antibacterial activities. The MICs of oxacillin for each of methicillin-resistant Staphylococcus aureus strains were reduced between 4 and 8-fold when these strains were coincubated with sub-MIC (≤ ½ MIC) levels of these compounds, demonstrating that the combination had synergistic effect for all cases.
Conclusions: Cuspidaria convoluta contains important pharmacologically active substances that can be used to improve antibiotic efficacy.
Objective: To predict B cell and T cell epitopes of 22-kDa, 47-kDa, 56-kDa and 58-kDa proteins.
Methods: The sequences of 22-kDa, 47-kDa, 56-kDa and 58-kDa proteins which were derived from Orientia tsutsugamushi were analyzed by SOPMA, DNAstar, Bcepred, ABCpred, NetMHC, NetMHC II and IEDB. The 58-kDa tertiary structure model was built by MODELLER9.17.
Results: The 22-kDa B-cell epitopes were located at positions 194-200, 20-26 and 143-154, whereas the T-cell epitopes were located at positions 154-174, 95-107, 17-25 and 57-65. The 47-kDa protein B-cell epitopes were at positions 413-434, 150-161 and 283-322, whereas the T-cell epitopes were located at positions 129-147, 259-267, 412-420 and 80-88. The 56-kDa protein B-cell epitopes were at positions 167-173, 410-419 and 101-108, whereas the T-cell epitopes were located at positions 88-104, 429-439, 232-240 and 194-202. The 58-kDa protein B-cell epitopes were at positions 312-317, 540-548 and 35-55, whereas the T-cell epitopes were located at positions 415-434, 66-84 and 214-230.
Conclusions: We identified candidate epitopes of 22-kDa, 47-kDa, 56-kDa and 58- kDa proteins from Orientia tsutsugamushi. In the case of 58-kDa, the dominant antigen is displayed on tertiary structure by homology modeling. Our findings will help target additional recombinant antigens with strong specificity, high sensitivity, and stable expression and will aid in their isolation and purification.
Acacia auriculiformis A.Cunn. ex Benth. is a perennial shrub having a wide range of medicinal potentials and is widely distributed throughout the world. It is being used traditionally to overcome various medical complications like sore eyes, aches, rheumatism, allergy, itching, and rashes. Besides, Acacia auriculiformis has been proven for many pharmacological activities like central nervous system depressant activity, antioxidant, antimicrobial, antimalarial, anti-filarial, cestocidal, antimutagenic, chemopreventive, spermicidal, wound healing, hepatoprotective and antidiabetic activity due to its low toxicity (LD50 = 3 741.7 mg/kg) and high efficacy. In addition, various phytochemical investigations reveal the presence of chief constituents as flavonoids (Auriculoside) and triterpenoid saponin glycosides (acaciasides- acaciaside A & B) in different parts of this plant. Since many years researchers have been carrying out various studies on this medicinal important shrub to elicit the various biological activities. This review attempts to highlight the pharmacognostical, phytochemical and pharmacological observations from 1965 to 2018 retrieved from SciFinder, Scientific journals, books, Google Scholar, and botanical electronic database websites. The various plant extracts evaluated for different pharmacological activities showed significant efficacy. Bioactive phytoconstituents isolated from various parts of the plant are highlighted. Pharmacognostical standardization of the plant done with various standard parameters is also reported. The low toxicity of this plant and the presence of major bioactive phytoconstituents like flavonoids and triterpenoid saponin glycosides are responsible for a therapeutic remedy for various diseases and pharmacological activities respectively. This review provides exhaustive information about the pharmacognostical, phytochemical, and pharmacological investigations of Acacia auriculiformis till date.
Authors: Francisca Sâmara Muniz dos Santos, José Weverton Almeida Bezerra, Jean Paul Kamdem, Aline Augusti Boligon, Marli Matiko Anraku, Ana Raquel Pereira da Silva, Kleber Ribeiro Fidelis, Nadghia Figueiredo Leite, Antônio Ivanildo Pinho, Henrique Douglas Melo Coutinho, José Edilson Gonçalves dos Santos
Objective: To evaluate the antibacterial activity and neuroprotective capacity of the ethanolic and aqueous extracts of Tarenaya spinosa (T. spinosa) as well as to determine and quantify some of its polyphenols by high performance liquid chromatography with diode-array detection (HPLC-DAD). Methods: The bacterial Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa strains, grown in Heart Agar Infusion, were tested. The drugs gentamicin, norfloxacin and imipenem were used to evaluate the modulating or antagonistic capacity of the T. spinosa extracts. The extract was analysed by HPLC-DAD to determine the main phenolic compounds. For the cell viability tests, individual heads of the Nauphoeta cinerea arthropod model were removed, homogenized in Trifluoromethyl ketone and centrifuged afterwards. Subsequently, 20 μL of NaNO2 were added to the biological material, except in the control group, to evaluate the protection capacity of the extracts. The homogenate of the insect heads was incubated for 2 h in tubes containing tetrazolium bromide. Results: HPLC-DAD demonstrated that the ethanolic extract of T. spinosa presented caffeic acid as the major compound. The ethanolic extract also showed neuroprotective effects at concentrations ≥ 10 μg/mL, while aqueous extract was shown to have a protective effect only at the concentration of 100 μg/mL. The aqueous extract demonstrated a clinically relevant antibacterial activity against the Staphylococcus aureus multidrug resistant strain - MDR, with MIC 512 μg/mL. However, when the extracts were associated with gentamicin and imipenem, a synergism was detected against Staphylococcus aureus and Escherichia coli MDR strains. Conclusions: Although it does not present an antibacterial action, the extracts of T. spinosa can be used in the pharmaceutical industries since its extracts show modulating action of drugs. Besides, these natural products have neuroprotective capacity.
Objective: To examine the in vitro and in vivo anti-inflammatory effects of the alkaloid enriched extract (ELA) from the roots of Eurycoma longifolia. Methods: The in vitro antiinflammatory effects of ELA were evaluated by examining its inhibitory activities against nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) expressions in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The level of NO produced in the culture media was determined by Griess method. The iNOS and COX-2 protein expressions were analyzed by Western blot. The in vivo effect of ELA was evaluated on LPS-induced septic shock in mice model. Mice mortality was monitored for 5 days after injection of LPS. The chemical contents of the ELA were determined by using various chromatographic and spectroscopic techniques. Results: The ELA was found to exhibit a significant anti-inflammatory effect in both in vitro and in vivo models. The results demonstrated that ELA dose-dependently inhibited LPS-induced NO production as well as the protein iNOS and COX-2 expressions. In the septic shock model, ELA dose-dependently protected mice from LPS-induced mortality. Further study on the isolated components of ELA indicated that 9,10-dimethoxycanthin-6-one may contribute significantly to the anti-inflammatory effects of the extract. Conclusions: These results suggest that ELA exhibits the anti-inflammatory activity via suppression of pro-inflammatory mediators such as NO, iNOS, and COX-2 and protects mice from LPS-induced mortality in septic shock model.
Objective: To investigate the in vitro antiproliferative action of essential oil from Salvia officinalis L. (S. officinalis) grown in Sicily (Italy), and its main components on hormone-dependent cancer cell lines. Methods: S. officinalis essential oil was prepared by hydrodistillation. The actions of the S. officinalis essential oil and its three principal components ( α -thujone, 1,8-cineole and camphor) were evaluated in LNCaP cells (prostate carcinoma), MCF7 cells (breast carcinoma) and HeLa cells (cervical carcinoma) at various dosages and diverse time points. Cell viability and proliferation were estimated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: S. officinalis essential oil at doses of 100 μg/mL and 200 μg/mL induced a significant reduction of cell viability in MCF7, LNCaP and HeLa cell lines after a 48-hour incubation. The same cell lines also showed decreased cell viability when they were treated with a mixture of three major components of the essential oil, at doses of 100 μg/mL and 200 μg/mL, after a 48-hour incubation. Conclusions: These preliminary results could shed light on the formulation of new therapeutic agents with antiproliferative activity. Thus supplementary investigations are fundamental to examine the molecular mechanisms of the anticancer effects of this species of Salvia in cancer cells and to achieve confirmation of its in vivo anticancer activity.
Objective: To investigate the efficacies of 12 essential oil (EO) formulations from three Zingiberaceae plants (Alpinia galanga, Curcuma zedoaria, and Zingiber cassumunar) individually and in combination with an augmenting Eucalyptus globulus (E. globulus) EO against females of Aedes albopictus (Ae. albopictus) and Anopheles minimus (An. minimus). Methods: These formulations were evaluated for their ovicidal, oviposition deterrent and adulticidal activities against Ae. albopictus and An. minimus by a topical method, a double-choice method and a WHO susceptibility test, respectively. Results: It was found that all formulations of Zingiberaceae plants EOs augmented with E. globulus EO were more effective in oviposition deterrent, ovicidal, and adulticidal activities against the two mosquito species than all of the formulations used without E. globulus EO. Their oviposition deterrent, ovicidal and adulticidal activities were equivalent to those of 10% w/v cypermethrin. In contrast, 70% v/v ethyl alcohol as a control alone was not effective at all. The highest synergistic effect in effective repellency against Ae. albopictus was achieved by 5% Alpinia galanga EO + 5% E. globulus EO and against An. minimus was 5% Zingiber cassumunar EO + 5% E. globulus EO. Moreover, the highest synergistic effects in ovicidal activities against Ae. albopictus and An. minimus were achieved by 10% Zingiber cassumunar EO + 10% E. globulus EO and 5% Curcuma zedoaria EO + 5% E. globulus EO, respectively. For the adulticidal activities, the highest synergistic effect against two mosquitoes was achieved by 5% Curcuma zedoaria EO + 5% E. globulus EO. Conclusions: These results suggest that Zingiberaceae plant EOs augmented with E. globulus EO have a high potential to be developed into oviposition deterrent, ovicidal, and adulticidal agents for controlling populations of Ae. albopictus and An. minimus.
Objective: To determine the role of toll-like receptor 4 (TLR-4) in eliciting cellular and humoral immune responses against recombinant Mycobacterium bovis bacille Calmette-Guérin (rBCG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum. Methods: Six groups of mice (n=6 per group) were injected with phosphate buffered saline T80, BCG or rBCG intraperitoneally, in the presence or absence of a TLR-4 inhibitor; TAK-242. Enzyme-linked immunosorbent assay was carried out for serum total IgG, IgG1, IgG2a and IgG2b determination. Spleens were also harvested and splenocytes cultured for determination of intracellular cytokines; IL-4 and IFN-γ via enzyme-linked immunosorbent assay. Results: The production of total IgG, and the subclasses IgG1, IgG2a and IgG2b was significantly higher in rBCG-immunised mice than BCG and phosphate buffered saline immunised mice in the absence of TAK-242. A significant rise in total IgG occurred with more booster immunisations. The level of IgG2a was highest, followed by IgG2b, then IgG1. The production of both IL-4 and IFN-γ was also highest in the rBCG immunised groups. These significant rises were inhibited in the presence of TAK-242. Conclusions: We present evidence of the role of TLR-4 in the increased production of total IgG, IgG1, IgG2a and IgG2b, as well as IL-4 and IFN-γ in response to our rBCG construct.
Objective: To determine spatial distribution of sand flies (Diptera: Psychodidae; Larroussius group), the vectors of visceral leishmaniasis in Ardabil province, Northwest of Iran. Methods: Sand flies were collected using sticky traps from the 30 selected points in Ardabil province, during May-November 2017. The MaxEnt model in GIS software was used for modeling. Results: A total of 2 794 specimens of sand flies were collected, of which 33% were Larroussius subgenus sand flies. Phlebotomus kandelakii and Phlebotomus wenyoni were the highest and lowest collected species respectively. Based on the modeling, four areas in the province were identified with more than 70% probability of the presence of Larroussius group vectors which were at risk of visceral leishmaniasis disease transmission. Conclusions: The distribution of Larroussius subgenus sand flies was observed in all parts of Ardabil. But the northern parts of the province (Germi and Bilesavar counties) as well as central part (Ardabil and Meshkinshahr counties) were of great importance in terms of the presence of Larroussius subgenus sand flies and the possibility of transmission of the visceral leishmaniasis.
Objective: To investigate the maturity development of miracidia in uterine eggs from four portions of the Opisthorchis viverrini uterus and environmental factors possibly affected in maturation and infectivity of distal part uterine eggs. Methods: Uteri of adult worms were divided into 4 equal parts. Development of eggs was determined under light microscope. Only embryonated eggs were used to evaluate the effects of physicochemical factors: temperature, salinity, acidity, ultraviolet A, B, C. Infection success was evaluated by feeding treated eggs to intermediate host snails and determining by using a PCR approach. Results: Eggs obtained from the uterus closest to the ovary (regions 1 and 2) failed to develop in vitro. Eggs from region 4 of the uterus (close to the genital pore) were used to study effects of physicochemical factors. The highest survival and infection success was in groups of eggs kept at 30 °C (95.20%). The calculated period of loss infection success (LI50 and LI95) on miracidia in distal uterine eggs by exposure to UV-A, UV-B and UV-C were 73 and 1 523 d; 8 and 20 d; 1 and 2 d, respectively. Lethal concentrations (LC50 and LC95) of salinity, HCl and NaOH on miracidia in distal uterine eggs were 45.43 and 120.09 ppt, 0.01 and 0.25 M; 0.01 and 0.11 M, respectively, after 24 h exposure. Conclusions: Opisthorchis viverrini eggs display a high tolerance to environmental conditions, especially after snail host eating for infection.
Objective: To identify mutually regulated proteins in PC-3 and DU145 androgen-independent prostate cancer cell lines treated with 1,5-bis(2-hydroxyphenyl)-1,4-pentadiene-3-one (MS17), and to study the molecular pathways that contributed to the anticancer activity of MS17. Methods: PC-3 and DU145 cells were treated with 3 × EC50 (15 μM) concentration of MS17 for 24 h and were subjected to protein expression profiling using two-dimensional gel electrophoresis and protein identification by mass spectrometry. Selected differentially expressed proteins with significant P-value of P<0.05 and fold change over 1.5-folds were filtered through and ontologically classified. Mutually regulated proteins were ranked by fold change and identified as common protein targets of MS17. Results: Profiling data revealed that, the mutually down-regulated proteins included ACTB and ACTG associated with structural molecule activity, ACTN1 with cell cycle, ACTN4 with cell migration, HNRPK with apoptosis, PLST with morphogenesis and TERA with proteolysis. However, the expressions of CH60 and HS71A respectively associated with response to unfolded protein demonstrated opposing regulation in PC-3 and DU145 cells. Pathway analysis of the differentially expressed proteins in PC-3 cells demonstrated the modulation of top pathways associated with cell-cell adhesion and cytoskeletal organization while in DU145 cells the pathways were associated with proteosomal degradation, regulation of electrolytes and water, regulation control of germ cells and organization of filament assembly/disassembly. Conclusions: The findings of the present study provide an understanding on the anti-tumorigenic activity of MS17 at the proteome level and warrant further research for its potential application for the management and treatment of androgen-independent prostate cancer.
Objective: To assess the safety and efficacy of herbal formulation rich in standardized fenugreek seed extract (IND-2) add-on therapy in type 2 diabetes mellitus (T2DM) patients who were on insulin treatment in prospective, single arm, open-label, uncontrolled, multicentre trial. Methods: T2DM patients (n=30) with aged 18-80 years who were stabilized on insulin treatment with fasting blood sugar (FBS) level between 100-140 mg/dL received IND-2 capsules (700 mg, thrice a day) for 16 weeks. The primary endpoints were an assessment of FBS at week 2, 4, 6, 8, 12 and 16. Secondary end-points include post-prandial blood sugar level, glycosylated Hb (HbA1c), reduction in the dose of insulin and number of hypoglycemic attacks, and improvement in lipid profile at various weeks. Safety and adverse events (AEs) were also assessed during the study. Results: Study was completed in twenty T2DM patients, and there was no significant reduction in FBS and post-prandial blood sugar level after addon therapy of IND-2. However, add-on therapy of IND-2 significantly reduced (P<0.01) the HbA1c values, requirements of insulin and hypoglycemic events as compared with baseline. Total cholesterol, high-density lipoproteins-cholesterol, and low-density lipoprotein-cholesterol levels were significantly increased (P<0.01) after IND-2 add-on therapy. Body weight and safety outcomes did not differ significantly in IND-2 add-on therapy group at week 16. Additionally, add-on therapy of IND-2 did not produce any serious adverse events. Conclusions: The results of present investigation suggest that add-on therapy of IND-2 with insulin in T2DM patients improves glycaemic control through a decrease in levels of HbA1c and number of insulin doses needed per day without an increase in body weight and risk of hypoglycemia. Thus, IND-2 may provide a safe and well-tolerated add-on therapy option for the management of T2DM.
Objective: To evaluate the anticancer activity of crude acetone and water leaf extracts of Tulbaghia violacea on a human oral cancer cell line (KB). Methods: The antioxidant activity of the leaf extracts was evaluated by using the DPPH assay while the anti-proliferative activity was assessed by using the MTT assay. The morphological characteristics of apoptotic cells were examined by using the dual acridine orange/ethidium bromide staining. Flow cytometry was used to evaluate the induction of multi-caspase activity and changes in the cell cycle. Results: The acetone and water extracts exhibited antioxidant activity in a concentration dependent manner. The extracts inhibited the growth of the KB cell line with IC50 values of 0.2 mg/mL and 1 mg/mL, respectively for acetone and water. Morphological changes such as cell shrinkage, rounding and formation of membrane blebs were observed in the treated cells. In acridine orange/ethidium bromide staining, the number of apoptotic cells increased as the concentration of the extracts increased. The activation of multi-caspase activity in KB cells treated with Tulbaghia violacea extracts was concentration dependent, leading to cell death by apoptosis and cell cycle arrest at the G2/M phase. Conclusions: The acetone and water extracts of Tulbaghia violacea appear to have anti-cancer activity against human oral cancer cells and need to be investigated further.
Authors: Dayane Kelly Dias do Nascimento Santos, Weslley Henrique de Oliveira Melo, Anastássia Mariáh Nunes de Oliveira Lima, Iranildo José da Cruz Filho, Gláucia Manoella de Souza Lima, Túlio Diego da Silva, Maiara Celine de Moura, Márcia Silva do Nascimento, Ana Maria Souto Maior, Thiago Henrique Napoleão, Cristiane Moutinho Lagos de Melo
Objective: To evaluate the structural and chemical composition of plant and the antioxidant and antimicrobial activities promoted by hexanic, ethanolic and ethyl acetate fractions obtained from leaves of Conocarpus erectus. Methods: Organic fractions were characterized through UPLC-MS and GC-MS. Antioxidant potential was performed through DPPH and molybdenum phosphate techniques. Antibacterial and antifungal assays were performed in accordance with Clinical and Laboratory Standards Institute protocols. Results: The obtained biomass of Conocarpus erectus leaves showed the high presence of glucose (0.45 g/L), cellulose (28.69%), Na (55.126 μg/L) and K (31.163 μg/L). We identified seven compounds in the hexanic and ethyl acetate fractions, and eight compounds in ethanolic fraction. Moreover, phenolic compounds are prevalent in all organic fractions with values of (10.04 ± 0.24), (221.26 ± 1.84), (340.53 ± 0.84) mg/g GAE to hexanic, ethyl acetate and ethanolic fraction, respectively. Antioxidant results showed a high potential in ethyl acetate fraction (71.82 ± 6.87)% and (10.89 ± 0.05)% in DPPH and molybdenum phosphate techniques, respectively. The ethanolic fraction showed moderate bacteriostatic and bactericidal activity against Staphylococcus aureus and presented a high fungistatic potential for all Candida species tested. Conclusions: Organic fractions obtained from leaves of Conocarpus erectus present antimicrobial and antioxidant properties, and these findings contribute to scientific information for the effectiveness on use of this plant in the development of a phytotherapic compound.
Objective: To investigate biomolecular alteration of sesamol on human lung adenocarcinoma (SK-LU-1) cells compared with cisplatin using Fourier transform infrared microscopy (FTIR). Methods: Cytotoxicity of sesamol was investigated against SK-LU-1 cells by using neutral red. DNA fragmentation and the cell cycle analysis were determined by agarose gel electrophoresis and flow cytometry, respectively. The FTIR microscopy technique was applied to explore the changes in cellular biochemical compositions in cells treated with sesamol that the biochemical and biological assays cannot cover. The alkylating property was determined by 4-(4-nitrobenzyl)pyridine assay. Results: Sesamol and cisplatin exerted an antiproliferative effect at 48 h with respective IC50 values of 2.7 and 0.07 mM. Both induced apoptosis by causing DNA damage and accumulation of cell populations at sub-G1. FTIR microscopy and Principle Component Analysis clearly discriminated the sesamol- and cisplatin-treated cells from the untreated cells or control. A significant increase of total lipid content was found in cisplatin-treated cells. Conformational changes in the proteins secondary structure from the α -helix to the β -sheet were found in both sesamol- and cisplatin-treated cells, as well as significant reductions in relative DNA content of both compared to the control were observed, suggesting DNA damage. A shift in the peak position of DNA region provides insight on the DNA interactions. Conclusions: The non-alkylating effect of sesamol based on the nitrobenzyl pyridine assay delineates the non-covalent binding mode of sesamol on DNA. Hydrogen bonding is the binding mode of sesamol on DNA, while for cisplatin it was covalent and hydrogen bonding.
Objective: To investigate the effect of crocin carotenoid on BNDF and CREB gene expression in the brain ventral tegmental area (VTA) and the serum level of BDNF in morphine-treated rats compared to control. Methods: In this study, 40 male Wistar rats (200-250 g) were used in 5 experimental groups: 1) non morphine treat rats (control); 2) non morphine-treated rats with 25 mg/kg crocin carotenoid (i.p., for 21 d); 3) morphine treated rats (10 mg/kg twice a day, s.c., 21 d); 4 and 5) morphine-treated rats with 12.5 and 25 mg/kg crocin carotenoid, respectively. By the end of research, BDNF and CREB expression was determined by real-time-PCR method. ELISA analysis was also applied for assessing the serum BDNF level. Results: The data indicated that morphine treatment could cause a significant decrease in BDNF and CREB gene expression (P<0.01 and P<0.001, respectively) in brain VTA as well as serum level of BDNF (P<0.01) in comparison to control group. Treatment with 25 mg/kg crocin carotenoid caused a significant enhancement in BDNF and CREF gene expression (P<0.01 and P<0.05, respectively) and serum level of BDNF (P<0.01) in morphine-treated rats in comparison to morphine-treated group. Conclusions: Regarding to obtained results, crocin carotenoid can inhibit unfavorable effects of morphine on the neural system to some extent through enhancing BDNF and CREB gene expression in brain VTA and serum level of BDNF.
Objective: To identify the bioactive extracts from Alternanthera sessilis and investigate its cytotoxicity potential against colon cancer cells, HT-29. Methods: This study examined the effects of three parts (aerial, leaf, stem) of whole plant on HT-29 colon cancer cell lines. Three different extracts from the plant parts were prepared by maceration technique using 80% ethanol. The anticancer activities were determined using MTT, clonogenic, cell motility and AOPI assay. The chemical composition profiling was analyzed by GC-MS. Results: Among three plant part extracts, leaf extract greatly suppressed the growth of colon cancer cells in time and dosage-dependent manner, followed by aerial and stem. The cytotoxicity results were rationalized with clonogenic, cell motility and AO/PI assay, where extract showed the most active activity compared to aerial and stem extracts. GC-MS analysis of leaf extract showed there were various recognized anti-cancer, anti-oxidant and anti-inflammatory compounds. Conclusions: Amid the screened extracts, the leaf extract exhibits the credible cytotoxic, anti-proliferative and apoptotic activity and hence, our findings call for additional research to conclude the active compounds and their mechanisms determining the apoptotic activity.
Objective: To investigate the possible protective and/or therapeutic potentials of Dunaliella salina (D. salina) biomass, its carotenoid and polar fractions on cardiac dysfunction associated with D-galactose (D-GAL) induced aging in rats. Methods: Aging associated cardiac dysfunction was induced in rats by injection of D-GAL (200 mg/kg; i.p) for 8 weeks. D-GAL injected rats were treated with two regimens; protective regimen where D. salina biomass (250 mg/kg), its carotenoid (250 μg/kg) and polar (250 μg/kg) fractions were given orally for two weeks concurrently with D-GAL injection as well as treatment regimen where the three treatments were given orally for 28 consecutive days after D-GAL injection. Results: D-GAL injection for 8 weeks was accompanied with dramatic electrocardiographic changes as well as profound elevation in serum levels of homocysteine, creatinine kinase isoenzyme and lactate dehydrogenase in addition to the reduction of the cardiac content of glucose trasporter 4. D-GAL also induced reduction in cardiac superoxide dismutase activity and elevation of inducible nitric oxide synthetase and interleukin-6. On the other hand, oral administration of D. salina carotenoid fraction as well as the total biomass significantly attenuated the D-GAL-induced disturbances in the above mentioned parameters where the protective regimen appeared more successful in controlling the manifestations of cardiac dysfunction. The histopathological examination further emphasized the promising results. Besides, the HPLC analysis of the carotenoid fraction of D. salina revealed the presence of 2.31% β -carotene. Conclusions: D. salina carotenoid fraction as well as the total biomass ameliorate D-GAL-induced aging associated cardiac dysfunction which is attributed to the potent antioxidant activity of β -carotene.
Objective: To investigate anti-quorum sensing (anti-QS) and anti-biofilm formation (anti- BF) activities of the ethanol extracts of 388 plants. Methods: The anti-QS activity of the plant extracts was evaluated by disc-diffusion assays using the bio-reporter strain, Chromobacterium violaceum CV017. Pseudomonas aeruginosa PAO1, Yersinia enterocolitica ATCC 9610, and Agrobacterium tumefaciens C58, which possess QS systems, were used to evaluate the anti- BF activity of the plant extracts. Results: Among 388 plant extracts, the Cornus controversa (C. controversa) and Cynanchum wilfordii extracts exhibited the strongest anti-QS activity. The C. controversa extract exhibited anti-BF activity against Pseudomonas aeruginosa, Yersinia enterocolitica and Agrobacterium tumefaciens, whereas the Cynanchum wilfordii extract exhibited no anti-BF activity against Pseudomonas aeruginosa. In addition, the C. controversa extract suppressed soft rot of cabbage. Conclusions: The C. controversa extract inhibits bacterial QS and BF, and is capable of controlling soft rot. Therefore, this extract has potential for the prevention and treatment of bacterial infections and for the development of alternatives to antibiotics.
Objective: To undertake metabolite profiling of various plant parts of Citrullus colocynthis, and assess antioxidant and wound healing activities of fractions for therapeutical applications. Methods: Extracts from leaves, stem, root, fruit pulp and seeds were analyzed using gas chromatography-mass spectrometry and high performance liquid chromatography. Variation in antioxidant potential was assayed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay. The extract with highest antioxidant potential was subjected on in-vivo wound healing activity using excision wound model. Results: Metabolite profiling of Citrullus colocynthis identified 70 chemically diverse metabolites from different plant parts by using a combination of GC-MS and HPLC. Concentration of colocynthin, a principal active secondary metabolite, ranged from 3.15 mg/g dry weight to 242.00 mg/g dry weight, the lowest being in leaves and highest in fruit pulp. DPPH radical scavenging activity of free radical (IC50) ranged from 196.44 μg/mL in fruit pulp to 413.33 μg/mL in leaves tissues. Significant wound contraction and increase in hydroxyproline content of granulation tissue were observed with ointment formulated from methanolic extract of fruit pulp. Conclusions: The study indicates that the methanol extract of Citrullus colocynthis fruit pulp when applied topically may promote wound contraction in rat model attributable to the accumulation of colocynthin. The high quantity of colocynthin (242.00 mg/g dry weight) and substantial concentration of 2,4-di-tert butyl phenol (3.2%), squalene (4.2%) and δ -tocopherol (2.5%) make this plant to provide new opportunities for development of medicinal, nutraceutical and dietary supplements with optimized functionality.
Objective: To investigate the role of toll-like receptor 2 (TLR2) in inflammatory activity of macrophage infected with the recombinant Mycobacterium bovis bacillus Calmette-Guerin (rBCG). Methods: Mouse macrophage cell line J774A.1 was infected with Mycobacterium bovis bacillus Calmette-Guerin (BCG) and rBCG cultures for 48 h in the presence or absence of 10 μg/mL of TLR2 inhibitor. Untreated macrophages were used as a negative control while lipopolysaccharide-stimulated macrophages were used as a positive control. The ability of the macrophage to engulf the BCG and rBCG in the absence or presence of TLR2 inhibitor was assessed using a phagocytic assay, while the production of inflammatory cytokines and nitric oxide by the infected macrophages was evaluated using ELISA and Griess reagent method, while the expression of the inducible nitric oxide synthase was determined using Western blot analysis. Results: The results showed that blocking TLR2 function reduced the phagocytic activity, nitric oxide production and proinflammatory cytokine secretion such as TNF- α, IL-1 β and IL-12p40 as well as inducible nitric oxide synthase expression in the infected macrophages. These data showed the importance of TLR2 in the activation of macrophages following BCG and rBCG infections. Conclusions: Through exploring the immunological mechanism which underlies the protection conferred by the candidate vaccine, this study will improve our understanding of the vaccine candidate’s mechanism to protect the host from malaria infection.
Objective: To evaluate effects of docosahexaenoic acid-enriched phosphatidylcholine (DHA-PC) on cytokine production induced by lipopolysaccharide (LPS). Methods: The culture supernatants of splenocytes exposed to DHA-PC along with LPS were harvested to determine the production of Th 1 (IFN- γ and IL-2) and Th2 [IL-4, IL-5, IL-6 and IL-12/IL-23(p40)] cytokines. Cytokines were measured using ELISA. Results: Co-administration of DHA-PC with LPS resulted in significantly lower IL-2 expression compared to that observed with administration of only LPS (P<0.01). Treatment with DHA-PC and LPS significantly increased IL-5 expression (P<0.01). Moreover, co-administration of DHA-PC with LPS significantly decreased IL-6 and IL-12/IL-23(p40) expressions compared to that observed with administration of only LPS (P<0.01). Conclusions: Our results suggest that DHA-PC inhibits pro-inflammatory cytokines [IL-2, IL-6 and IL-12/IL-23(p40)] expression on induction of inflammation.
Authors: Paulo Fernando Machado Paredes, Selene Maia de Morais, Fernando César Rodrigues Brito, Luiz Francisco Wemmenson Gonçalves Moura, Patrícia de Araújo Rodrigues, Stephen Rathinaraj Benjamin, Francisco Ernani Alves Magalhães, Eridan Orlando Pereira Tramontina Florean, Maria Izabel Florindo Guedes
To evaluate the chemical components of active extract from Cnidoscolus quercifolius root bark and its cytotoxic potential against several tumor strains. Methods: The high-performance liquid chromatography with diode-array detection and 1H and 13C nuclear magnetic resonance spectroscopy of the extract were used to distinguish the existence of possible functional groups in the root bark extract. The in vitro cytotoxic activity of methanol extract on human colon cancer cell lines was evaluated using OVCAR-8, SF-295, HCT-116, HL-60 strains and the samples were assessed by 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide method. Results: The analysis of nuclear magnetic spectra of the active chloroform fraction revealed the presence of absorptions bands correspondent to a mixture of favelines such as neofavelanone, deoxofaveline or methyl-faveline, which structures were confirmed by ultraviolet spectra upon high-performance liquid chromatography with diode-array detection analysis. The active fraction showed cytotoxic effects in the tested strains, HCT-116, SF-295, OVCAR-8 and HL-60 cells with IC50 of 72 hours ranging from 4.95 to 15.23 μg/mL. Conclusions: The results suggest that the substances present in faveleira (Cnidoscolus quercifolius) root bark extract have a cytotoxic potential against several tumor lines, showing a broader antitumour potential, and in addition no adverse to healthy cells. Therefore, the root bark extract of Cnidoscolus quercifolius has a possibility of use for anticarcinogenic therapies.
Objective: To evaluate the protective effect of morin against pentylenetetrazol (PTZ)-induced tonic-clonic convulsions in mice. Methods: Swiss albino mice (18-22 g) was used to induce convulsions by intraperitoneal (i.p.) administration of PTZ (90 mg/kg). Mice were either pretreated with morin (10, 20 and 40 mg/kg) or vehicle (distilled water, 10 mg/kg) 45 min before PTZ administration. Various behavioral and biochemical parameters were assessed. Results: PTZ administration resulted in significant production (P<0.001) of tonic-clonic conclusion and mortality in mice. PTZ-induced increase in the duration of convulsion, onset of convulsion and mortality was inhibited significantly by morin (20 and 40 mg/kg) administration. The PTZ-induced decrease in brain GABA, dopamine and Na+K+ATPase levels and increase in xanthine oxidase activity were inhibited significantly by morin (20 and 40 mg/kg) treatment. The increased levels of malondialdehyde and nitric oxide level were significantly decreased by morin (20 and 40 mg/kg) treatment. Also, reduced levels of superoxide dismutase and glutathione were increased significantly by morin treatment. Conclusions: Results of the present study indicate that morin showed its anti-convulsant effect via modulating the levels of brain GABA, Na+K+ATPase, and oxido-nitrosative stress. Thus, morin can be a potential candidate for further clinical evaluations as an anti-epileptic agent.
Objective: To produce fluorescent tagged recombinant erythroferrone protein (ERFE_eGFP) for laboratory investigations. Methods: Erythroferrone (ERFE) gene was fused to green fluorescent protein (eGFP) gene and cloned in a pSecTag2Hygro plasmid. The constructed plasmid was amplified in Escherichia coli DH5 α and the eGFP-fused ERFE (ERFE_eGFP) protein was expressed in human embryonic kidney (HEK293T) cell line. Results: The plasmid constructed from colony C6 contained ERFE_eGFP with the correct restriction sizes of 4.2 kb and expressed secretory ERFE_eGFP fusion protein (approximately size of 75 kDa) in HEK293T cell line. Conclusions: ERFE_eGFP recombinant protein is successfully expressed as a secretory functional protein and could be sensitively detected using fluorometry. This fusion protein might benefit future applications for localization of cellular ERFE receptors and competitive immunoassay of ERFE concentration.
Objective:To identify the region conferring stability to pBSSB2 (a linear plasmid, pBSSB1,
containing a kanamycin cassette), which is unique to Indonesian isolates of Salmonella enterica
serovar Typhi. Methods:The open reading frame (ORF) 009 was identified as a toxin coding
gene in the plasmid through introduction of translational termination codons in the ORF.
Results:The stability function was located in a fragment that spanned nucleotides 5 766 to
6 828 in the linear plasmid genetic map. Ectopic expression of ORF009 in pBAD18 vector
indicated ORF009 codes for a toxin. This fragment could stabilize plasmid pUC18 previously
destabilized through mutation of the pcnB(plasmid copy number control) gene that codes
for polyA polymerase. Majority of the cells expressing ORF009 were non-viable according
to phase contrast microscopy. Conclusions:This study demonstrated that a linear plasmid
fragment that carries a gene encoding a toxin possibly conferred stability to the parent plasmid.
It was able to stabilize a multicopy plasmid of Escherichia coli.
Objective: To investigate different types of dates and medical properties of influencing blood clotting and wound healing in an animal model. Methods: Three different cultivars of dates (Ajwa, Khalas, and Fardh) were examined in-vivo, for blood clotting and wound healing using CD1 mice of both sexes. Study of toxicity to animals was performed accordingly prior to further investigations. The ethanolic extracts were given orally to animals as a constituent in their daily water. Blood samples were obtained from the mice inferior vena cava to carry out the prothrombin time (PT) assay using the manual method and confirmed using a semi-automated machine. The bleeding time (BT) assay was performed using the cutting technique. In the wound healing analysis, a small cut (5-10 mm) in the skin overlying the thigh was conducted in all mice under anesthesia. The diameter of the cut and healing status were measured on a daily basis throughout the time of the experiment using a roller. Results: Ajwa was able to elevate both PT and BT (P<0.05), significantly in a time-dependent manner followed by Khalas date (P<0.05). The results of PT and BT of Fardh date were found to be very close to those of the control group (P<0.05). Despite its activity as an anticoagulant, Khalas date showed a potential property to enhance wound healing in contrast to other dates and the control groups in this study. Conclusions: Omani Khalas date fruit has both antithrombotic as well as wound healing properties. The results open a new gate with these fruits for exploring the potential component(s) that may play an important role in antithrombotic as well as wound healing process.