|Quality assessment of pollen typhae by high-performance liquid chromatography fingerprint, hierarchical cluster analysis, and principal component analysis|
Xin Ma, Haimiao Zou, Yibin Pan, Jing Su, Yujiao Qiu, Mingfeng Qiu
Pharmacognosy Magazine 2019 15(61):177-182
Aim: This study aims to establish the quality assessment methods of Pollen Typhae. Materials and Methods: High performance liquid chromatography (HPLC) fingerprint analysis, hierarchical cluster analysis (HCA), and principal component analysis (PCA) were used for quality evaluation of Pollen Typhae from different origins together with microscopic identification. Then, the quantity of 43 crude Pollen Typhae samples in the market was collected and analyzed. Results: In true and false test, four False Pollen Typhae samples, 13 Net Pollen Typhae (NPT) samples, and 26 Grass Pollen Typhae (GPT) samples were identified by microscopic identification. In quality test, the amounts and percentages of Qualified Pollen Typhae, Unqualified Pollen Typhae were 24 (55.81%) and 19 (44.19%), respectively with typhaneoside and isorhamnetin-3-O-neohesperidoside determined by HPLC according to China Pharmacopeia. We analyzed 43 samples from 20 regions and established their fingerprints, then selected 31 peaks as characteristic peaks and calculated their relative peak areas. To express the HPLC fingerprints quantitatively, peak 16, 18, 22, 23, and 26 were verified as typhaneoside, isorhamnetin-3-O-neoheptanoside, rutin, quercetin, and isorhamnetin. The similarity of correlation coefficients in chromatogram was 0.954 ± 0.007 and 0.922 ± 0.004 for NPT and GPT, respectively, while 0.67 ± 0.008 for 43 samples. The analysis of HCA and PCA can distinguish true or false, qualified or unqualified of Pollen Typhae. Conclusion: HPLC fingerprint combined with HCA and PCA provides a very efficient and comprehensive method for quality evaluation of Pollen Typhae. Abbreviations used: HCA: Hierarchical cluster analysis; PCA: Principal component analysis; FPT: False Pollen Typhae; NPT: Net Pollen Typhae; GPT: Grass Pollen Typhae; QPT: Qualified Pollen Typhae; UPT: Unqualified Pollen Typhae; RSDs: The relative standard deviations; CASE: Computer Aided Similarity Evaluation; TCM: Traditional Chinese medicine.
|Chamuangone-enriched Garcinia cowa leaf extract with rice bran oil: Extraction and cytotoxic activity against cancer cells|
Pirunrat Sae-Lim, Supreeya Yuenyongsawad, Pharkphoom Panichayupakaranant
Pharmacognosy Magazine 2019 15(61):183-188
Background: Chamuangone has been isolated from Garcinia cowa leaves and exhibited various biological activities, i.e., antibacterial, anti-Leishmania major, and cytotoxic activity against cancer cells. n-Hexane has been reported to be the most suitable solvent for extraction of chamuangone. Objectives: Some vegetable oils were determined as an alternative green solvent for extraction of an anticancer compound, chamuangone from G. cowa leaf. The chamuangone-enriched extract was standardized and evaluated for cytotoxic activity against human cancer cell lines. Materials and Methods: Microwave-assisted extraction and high-performance liquid chromatography were used for extraction and standardization. The cytotoxic activity was determined using a sulforhodamine B assay. Results: The chamuangone-enriched extract was obtained using rice bran oil as the alternative green solvent and standardized to contain 1.97 mg/mL chamuangone. The extract exhibited cytotoxic activity against human lung adenocarcinoma, human breast adenocarcinoma, and human colorectal adenocarcinoma cell lines, with IC50 values of 15.3, 15.9, and 12.8 μg/mL, respectively, but was nontoxic to human gingival fibroblasts, a normal cell line, at a concentration of 50 μg/mL. Moreover, the extract contained several natural antioxidants, including α-tocopherol (76.7 mg/100 g), γ-oryzanol (cycloartenol ferulate: 67.1 μg/mL and 24-methylenecycloartanol ferulate: 85.6 μg/mL), and antioxidant capacity determined as ascorbic acid (258.7-mM ascorbic acid equivalent per gram). Conclusion: Based on these findings, the chamuangone-enriched extract may be considered as a novel functional food in cancer chemopreventive action. Abbreviations used: AAE/g: Ascorbic acid equivalent per gram; CEO: Chamuangone-enriched Garcinia cowa leaf extract with rice bran oil; DMEM: Dulbecco's Modified Eagle Medium; FBS: Fetal bovine serum; HGF: Human gingival fibroblasts; HPLC: High-performance liquid chromatography; IC50:50% Inhibitory concentration; MAE: Microwave-assisted extraction; MHz: Megahertz; MUFAs: Monounsaturated fatty acids; NMR: Nuclear magnetic resonance; ODS: Octadecylsilane; PUFAs Polyunsaturated fatty acids; SD: Standard deviation; SFAs: Saturated fatty acids; SRB: Sulforhodamine B; TCA: Trichloroacetic acid; UFAs: Unsaturated fatty acids; UV: Ultraviolet; W: Watt.
|Impact of ginsenoside-Rg3 on catecholamine secretion in the perfused model of the rat adrenal medullae|
Ki-Hwan Kim, Hyo-Jeong Lim, Young-Jae Ki, Dong-Yoon Lim
Pharmacognosy Magazine 2019 15(61):189-198
Background: The present study was the first attempt to explore the characteristics of ginsenoside-Rg3 (Rg3) on release of catecholamines (CA) in the perfused rat adrenal medullae and also to verify the underlying action mechanism. Materials and Methods: The adrenal medulla was separated by some modification of the previous method and perfused with Krebs solution. CA was assayed directly by the fluorometry. Results: Rg3 reduced acetylcholine (ACh)-produced CA release in a dose- and time-dependent manner. Rg3 time-dependently depressed CA release produced by 3-(m-chloro-phenyl-carbamoyl-oxy)-2-butynyltrimethyl ammonium chloride (McN-A-343), 1,1-dimethyl-4-phenyl piperazinium iodide, and angiotensin II. In the presence of Rg3, the CA release produced by high K+, veratridine, cyclopiazonic acid, and methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4- (2-trifluoromethyl-phenyl)-pyridine-5-carboxylate (Bay-K-8644) was also markedly suppressed. However, during the simultaneous perfusion of Rg3 and Nω-nitro-l-arginine methyl ester hydrochloride (l-NAME), the release of CA produced by ACh, angiotensin II, Bay-K-8644, and veratridine was restored closely to the level of each control, in contrast to that of Rg3-treatment alone. The nitric oxide (NO) release was significantly elevated by Rg3-treatment. Furthermore, in the coexistence of Rg3 and fimasartan, ACh-produced CA release was more significantly reduced as compared to that of fimasartan-treatment alone. Conclusions: We present the first evidence that Rg3 markedly depresses the CA secretion produced by activation of neuronal cholinergic and angiotensinergic receptors. Rg3-produced inhibition appears to be evoked not only by blocking the inflow of Na+ and Ca2+ into adrenomedullary cells but also by preventing the Ca2+ release from intracellular storage, partly through enhancement of NO release by NO synthase activation. Coadministration of Rg3 and fimasartan may be clinically beneficial for the treatment of cardiovascular diseases. Abbreviations used: Rg3: Ginsenoside-Rg3; VDCC: L-type voltage-dependent Ca2+ channels; Bay–K-8644: Methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4-(2-trifluoromethyl-phenyl)-pyridine-5-carboxylate; ACh: Acetylcholine; McN-A-343: 3-(m-chloro-phenyl-carbamoyl-oxy)-2-butynyltrimethyl ammonium; DMPP: 1.1-dimethyl-4-phenyl piperazinium; NO: Nitric oxide; L-NAME: Nω-nitro-l-arginine methyl ester; nNOS: Neuronal NO synthase.
|New bioactive C15 acetogenins from the red alga Laurencia obtusa|
Mohamed A Ghandourah, Walied M Alarif, Nahed O Bawakid
Pharmacognosy Magazine 2019 15(61):199-203
Background and Objective: With regard to the uniqueness of the red algae of the genus Laurencia as the source of C15-acetogenins, along with the diversity of biological applications; the acetogenin content of the Red Sea Laurencia obtusa was investigated. Materials and Methods: Fractionation and purification of the CH2Cl2/MeOH extract were carried out by applying several chromatographic techniques, including column and preparative thin-layer chromatography; followed by a series of 1H nuclear magnetic resonance measurements to give rise of some interesting notes. Toxicity to Artemia salina was evaluated. The apoptosis induced by these two compounds was demonstrated by DNA fragmentation assay and microscopic observation. Results: A new rare chloroallene-based C15 acetogenin, laurentusenin (1) along with a new furan ring containing C15 acetogenin, laurenfuresenin (2), were isolated from the red alga L. obtusa. Comparing 1D and 2D NMR, MS, ultraviolet and infrared radiation spectral data for the newly isolated compounds with the reported bromoallene containing acetogenins spectral data was played the crucial role for characterization of their chemical structures. 1 and 2 exhibited bare toxicity (LD50 >12 mM) in test organism, A. salina and induced apoptotic death confirmed by DNA fragmentation and microscopic investigations. Conclusion: The isolated metabolite 1 showed unusual substituted allene side chain, while 2 inserted furan ring as a new acetogenin nucleus. Both compounds may play a role in apoptosis induction and initiation and propagation of inflammatory responses. Abbreviations used: NMR: Nuclear magnetic resonance; MS: Mass spectrometry; UV: Ultraviolet spectroscopy; IR: infrared radiation; EIMS: Electron ionization mass spectra; TLC: Thin-layer chromatography; PPP: Platelet poor plasma; PRPDS: platelet-rich plasma derived serum. DEPT: Destortionless Enhancement by Polarization Transfer; NOESY: Nuclear Overhauser Effect Spectroscopy; HSQC: Heteronuclear Single Quantum Coherence; HMBC: Heteronuclear Multiple-quantum Correlation; 1H-1H COSY: Correlation Spectroscopy.
|A new antifungal aminobenzamide derivative from the endophytic fungus Fusarium sp.|
Sabrin R M. Ibrahim, Gamal A Mohamed, Maan T Khayat, Rwaida A Al Haidari, Amal A El-Kholy, Mohamed F Zayed
Pharmacognosy Magazine 2019 15(61):204-207
Background: Endophytic fungi attracted attention as a prolific source of bioactive natural products with a potent pharmaceutical activity and unique structure. Objective: The main goal of the study is to separate and identify the bioactive constituents from the endophytic fungus Fusarium sp. as well as to evaluate the antimicrobial of the new metabolites. Materials and Methods: The fungus was cultured on a rice medium, and then, the cultures were extracted with ethyl acetate (EtOAc). The EtOAc extract was chromatographed utilizing different chromatographic methods to give five metabolites. The structural determination of these metabolites was carried out by the analyses of various spectroscopic data, in addition to comparison with the formerly reported data. The antifungal and antibacterial potentials were evaluated toward various microbial strains using disc diffusion assay. Results: A new aminobenzamide derivative, namely fusaribenzamide A (2), and four known metabolites: (22E,24R)-stigmasta-5,7,22-trien-3-β-ol (1), adenosine (3), p-hydroxyacetophenone (4), and tyrosol (5) were isolated. Fusaribenzamide A (2) possessed significant antifungal activity toward Candida albicans with minimum inhibitory concentration (MIC) value 11.9 μg/disc compared to nystatin (MIC 4.9 μg/disc). Conclusion: The endophytic fungus Fusarium sp. could be considered as a wealthy pool for the isolation of aminobenzamide derivatives. Fusaribenzamide A may be a candidate for the discovery of a promising antifungal agent. Abbreviations Used: CC: Column chromatography; CHCl3: Chloroform; COSY: Correlations spectroscopy; DBE: double bond equivalent; EtOAc: Ethyl acetate; DMSO: Dimethyl sulfoxide; H2SO4: Sulfuric acid; HMBC: Heteronuclear multiple bond correlation experiment; HRMS: High-resolution mass spectrometry; HRESIMS: High-resolution electrospray ionization mass spectrometry; HSQC: Heteronuclear single quantum correlation; IR: Infrared; IZD: Inhibition zone diameter; KBr: Potassium bromide; LTQ: Linear trap quadrupole; MeOH; Methanol; MIC: Minimum inhibitory concentration; NMR: Nuclear magnetic resonance; RP: Reversed phase; SiO2: Silica gel; TLC: Thin-layer chromatography; UV: Ultraviolet; VLC: Vacuum liquid chromatography.
|Optimization of infrared-assisted extraction of bioactive lactones from Saussurea lappa L. and their effects against gestational diabetes|
Karim Raafat, Nada El-Darra, Fatima A Saleh, Hiba N Rajha, Nicolas Louka
Pharmacognosy Magazine 2019 15(61):208-218
Background: Saussurea lappa (S. lappa, Asteraceae) have immunomodulatory effects and used in the management of many metabolic disorders. Gestational diabetes is one of the metabolic disorders affecting globally one in seven pregnant women. Objectives: The aim of the current study is to optimize an infrared-assisted extraction (IR-AE) method for S. lappa bioactive constituents, phytochemically investigate its content, isolate its most active constituent, and to assess their biological effects against gestational diabetes. Materials and Methods: To optimize IR-AE conditions, four main factors were studied including solvent concentration, extraction time, powder size, and IR power in the yielded extract (SL-IR). Reversed-phase high-performance liquid chromatography coupled with bio-guided fractionation and isolation procedures using 1H and 13C NMR method were utilized. Solid–liquid (SL-SLE) and ultrasound (SL-US) extraction methods were also done. Results: The optimal IR-AE extraction conditions were found to be 20% aqueous phase concentration, 60-min extraction time, 70 mesh powder size, and 70 W IR power. Phytochemically, four major lactones were identified, including costunolide, dehydrocostuslactone, isoalantolactone, and alantolactone (ATL). ATL was the most active lactone. SL-IR, SL-US, SL-SLE, or ATL showed a significant (P < 0.05) and dose-dependent hypoglycemia in pregnant diabetic group, adequate fetus weight percentage elevation and did not show any external anomalies. The best control of gestational diabetes, insulin secretagogue potentials, elevation in serum catalase and reduced glutathione levels, and lipid peroxidation decrease were demonstrated by SL-IR 250 mg/Kg. The antioxidant and the insulin secretagogue activities might be among the main mechanisms, whereby the SL-IR controls gestational diabetes and decreases offspring anomalies. Conclusion: Currently, it is the first time to optimize an IR-AE method for extracting bioactive lactones from S. lappa. The optimized IR-AE technique has shown to be a rapid and efficient extraction method with SL-IR showing superiority in controlling gestational diabetes for pregnant groups coupled with high safety profile on the offspring. Abbreviations used: IR-AE: Infrared-assisted extraction; SL: Saussurea lappa, S. lappa; SL-IR: Saussurea lappa infrared extract; SL-US: Saussurea lappa ultrasound extract; SL-SLE: Saussurea lappa solid–liquid extract; ATL: Alantolactone; NDC: Nondiabetic control; DC: Diabetic control; MTF: Metformin; TBARS: Thiobarbituric acid; GSH: Reduced glutathione; CAT: Catalase; APA: Adequate for pregnancy age; LPA: Large for pregnancy age; SPA: Small for pregnancy age.
|Dictamnus dasycarpus Turcz., root bark alleviates oxazolone-induced atopy-like dermatitis in mice|
Beodeul Yang, Ji Hyo Lyu, Sura Kim, Young Chul Park, Koanhoi Kim, Hyungwoo Kim
Pharmacognosy Magazine 2019 15(61):219-225
Background: Dictamnus dasycarpus Turcz., is one of the most frequently used herbal medicine to treat dermatosis associated with psoriasis, pruritus, scabies, and eczema. Objective: We investigated the anti-inflammatory effects of D. dasycarpus extract (DDE) using a mouse model with atopic dermatitis (AD)-like dermatitis. Materials and Methods: The therapeutic effects of DDE on skin lesion, tone of color, and inhibitory effects on histopathological changes and cytokine production in skin tissues were assessed in mice with AD-like dermatitis induced by oxazolone. Results: Topical application of DDE alleviated skin lesions such as erythema, scaling and excoriations, and ameliorated erythema and the melanin index. In addition, DDE effectively prevented skin enlargement induced by oxazolone, while also preventing epidermal hyperplasia, spongiotic change, and hyperkeratosis and reducing the production of tumor necrosis factor-alpha and interleukin (IL)-4 and IL-5 in inflamed tissues. Finally, DDE did not affect changes in body weight and spleen–body weight ratio relative to dexamethasone. Conclusion: These results indicate that D. dasycarpus can be used as a topical agent for inflammatory skin diseases with relative safety. Abbreviation used: HM: Herbal medicine; DNFB: Dinitrofluorobenzene, ICAM-1: Intercellular adhesion molecule-1, AD: Atopic dermatitis, DDE: Dictamnus dasycarpus extract, AOO: Vehicle composed of acetone and olive oil, DEX: Dexamethasone, CBA: Cytometric bead array.
|Antidepressant-like effects of methanol extract and fractions of Hypericum juniperinum kunth in the forced swimming test|
Laura A Mejía-Agudelo, Maritza A Rojas, Mario F Guerrero-Pabón, Freddy A Ramos, Leonardo Castellanos, Juan Camilo Marín-Loaiza
Pharmacognosy Magazine 2019 15(61):226-231
Background: Some members of the genus Hypericum have been shown to demonstrate antidepressant-like effects. In Colombia, approximately 54 species of Hypericum have been reported, and only a few have been investigated chemical and pharmacologically. Objective: The aim of this study was to evaluate the antidepressant-like effects of a methanol extract, as well as ethyl acetate and butanol fractions, obtained from aerial parts of Hypericum juniperinum K. Materials and Methods: Behavioral and locomotor activities were evaluated in the open field test (OFT). Antidepressant-like activity was measured in the forced swimming test (FST) in male Swiss albino mice. Preliminary phytochemical screening as well as a high-performance liquid chromatography (HPLC) profile of the active fractions of H. juniperinum was performed. Results: Treatment with the methanol extract at 500 mg/kg and the ethyl acetate and butanol fractions at 150 and 300 mg/kg resulted in a decrease in the immobility times in FST. In the OFT, none of the treatments presented altered behavior or locomotor activity of mice. Preliminary phytochemical screening identified terpenes/steroids, flavonoids, phenol derivatives, tannins, and saponins. In the HPLC analysis of the fractions, rutin, quercitrin, and quercetin were identified with the help of coinjection of standards. Conclusion: This is the first report of the antidepressant-like activity of extracts and fractions obtained from H. juniperinum in the FST model of depression. Flavonoids may be responsible for the antidepressant-like action of H. juniperinum. Abbreviations used: FST: Forced swimming test; HJE: Methanol extract of Hypericum juniperinum; HJEAF: Ethyl acetate fraction obtained from the methanol extract of Hypericum juniperinum; HJEBF: Butanol fraction obtained from the methanol extract of Hypericum juniperinum; HPE: Methanol extract of Hypericum perforatum; HPLC: High-performance liquid chromatography; IMI: Imipramine; MeOH: Methanol; OFT: Open field test; tR: Retention time; UV: Ultra violet; VEH: Vehicle.
|Biological evaluation and molecular docking study of metabolites from Salvadora Persica L. Growing in Egypt|
Mohammed M Ghoneim, Wael M Afifi, Mohamed Ibrahim, Mohamed Elagawany, Maan T Khayat, Mohamed H Aboutaleb, Ahmed M Metwaly
Pharmacognosy Magazine 2019 15(61):232-237
Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a resistant staph bacterium to several antibiotics causing several lives-threating diseases such as pneumonia and sepsis. Meswak, Salvadora persica, exhibited promising antimicrobial properties before. Objective: Exploring the anti-MRSA activity of S. persica L. metabolites and its mechanism of action on a molecular level. Materials and Methods: Structure elucidation of the isolated metabolites was carried out by spectroscopic data (one-dimensional and two-dimensional nuclear magnetic resonance). The biological activities of the isolated metabolites against MRSA were evaluated and the molecular mode of action against the dehydrosqualene synthase enzyme have been done. Results: Four compounds have been isolated and identifies to be; apigenin (1), luteolin (2), astragalin (3), and kaempferol-3-O-rhamnoside (4). Compounds 1–4 showed good anti-MRSA activities with IC50 values of 10.3, 11.5, 3.5, and 4.5 μg/mL, respectively. In consistent, astragalin and kaempferol-3 rhamnoside showed close high docking scores. Herein, we are reporting the molecular determinates of activity of these new scaffolds as anti-MRSA, which would be of great importance to developing new anti-MRSA candidates. Abbreviations Used: 1D: One-dimensional; 2D: Two-dimensional; CC: Column chromatography; COSY: Correlations spectroscopy; DMSO: Dimethyl sulfoxide; HMBC: Heteronuclear multiple-bond correlation experiment; HRESIMS: High-resolution electrospray ionization mass spectrometry; HSQC: Heteronuclear single-quantum correlation; IR: Infrared; MRSA: Methicillin-resistant Staphylococcus aureus; NMR: Nuclear magnetic resonance; RP: Reversed phase; TLC: Thin-layer chromatography; UV: Ultraviolet; VLC: Vacuum liquid chromatography.
|Ameliorative effects of tannic acid on lipopolysaccharide-induced sepsis and acute lung injury in mice|
Ruirui Zhang, Minyan Dang, Shoutian Qiu, Hongyan Gu, Pingping He, Gang Guo, Tao Zhang
Pharmacognosy Magazine 2019 15(61):238-243
Background: Acute lung injury (ALI) caused by endotoxins is a severe complication causing lethal conditions. Prevention of the inflammatory response is necessary to overcome this condition. Tannic acid is a polyphenol known for its pharmacological properties such as antioxidant, antimicrobial, anti-inflammatory, anticancer, antitumor, and antimutagenic. This study aims to investigate the anti-inflammatory mechanism of tannic acid on lipopolysaccharide (LPS)-induced sepsis and ALI in mice. Materials and Methods: Male BALB/c mice were divided into five groups (n = 12) and induced with LPS (50 mg/kg body weight) and treated with tannic acid (25, 50, and 100 mg/kg) after 1 h. The bronchoalveolar lavage fluids were collected to determine the myeloperoxidase activity and levels of inflammatory cytokines (tumor necrosis factor-α, interleukin [IL]-6, and IL-1 β). Nuclear factor-kappa-β (NF-κβ) activities were examined through Western blot analysis, and hematoxylin and eosin staining was done for histopathological alterations of lung tissues. Results: Results of the study showed that tannic acid was able to prevent the infiltration of inflammatory cells and cytokines into the site of ALI which is also connected to the suppression of NF-κβ activation as shown in the Western blot analysis. The histopathological results further support these results. The safety of tannic acid was also proven on the survival of RAW264.7 cells. Conclusion: The anti-inflammatory mechanism of tannic acid on LPS-induced ALI and sepsis can be credited to the inhibition of inflammatory cytokines production mediated by NF-κβ pathway suppression. Abbreviations used: ALI: Acute lung injury; BALFs: Bronchoalveolar lavage fluids; NO: Nitric oxide; COX-2: Cyclooxygenase-2; iNOS: Inducible nitric oxide synthase; TNF-α: Tumor necrosis factor-α; IL-1β: Interleukin-1 β; IL-6: Interleukin-6.
Τετάρτη, 6 Μαρτίου 2019
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