Molecules

1161 KiB

Open AccessArticle

Antioxidant and Anticancer Activities of Essential Oil from Gannan Navel Orange Peel

by Chao Yang, Hui Chen, Hongli Chen, Balian Zhong, Xuzhong Luo and Jiong Chun

Molecules 2017, 22(8), 1391; https://doi.org/10.3390/molecules22081391 - 22 Aug 2017

Cited by 127 | Viewed by 14570

China is one of the leading producers of citrus in the world. Gannan in Jiangxi Province is the top navel orange producing area in China. In the present study, an essential oil was prepared by cold pressing of Gannan navel orange peel followed [...] Read more.

China is one of the leading producers of citrus in the world. Gannan in Jiangxi Province is the top navel orange producing area in China. In the present study, an essential oil was prepared by cold pressing of Gannan navel orange peel followed by molecular distillation. Its chemical composition was analyzed by GC-MS. Twenty four constituents were identified, representing 97.9% of the total oil. The predominant constituent was limonene (74.6%). The anticancer activities of this orange essential oil, as well as some of its major constituents, were investigated by MTT assay. This essential oil showed a positive effect on the inhibition of the proliferation of a human lung cancer cell line A549 and prostate cancer cell line 22RV-1. Some of the oil constituents displayed high anticancer potential and deserve further study. Full article

(This article belongs to the Special Issue Essential Oils: Chemistry and Bioactivity)

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1160 KiB

Open AccessArticle

The Potential of ?-Spinasterol to Mimic the Membrane Properties of Natural Cholesterol

by Ivan Haralampiev, Holger A. Scheidt, Daniel Huster and Peter Müller

Molecules 2017, 22(8), 1390; https://doi.org/10.3390/molecules22081390 - 22 Aug 2017

Cited by 5 | Viewed by 5668

Abstract

Sterols play a unique role for the structural and dynamical organization of membranes. The current study reports data on the membrane properties of the phytosterol (3?,5?,22E)-stigmasta-7,22-dien-3-?-ol (?-spinasterol), which represents an important component of argan oil and have not been investigated so far in [...] Read more.

Sterols play a unique role for the structural and dynamical organization of membranes. The current study reports data on the membrane properties of the phytosterol (3?,5?,22E)-stigmasta-7,22-dien-3-?-ol (?-spinasterol), which represents an important component of argan oil and have not been investigated so far in molecular detail. In particular, the impact of ?-spinasterol on the structure and organization of lipid membranes was investigated and compared with those of cholesterol. Various membrane parameters such as the molecular packing of the phospholipid fatty acyl chains, the membrane permeability toward polar molecules, and the formation of lateral membrane domains were studied. The experiments were performed on lipid vesicles using methods of NMR spectroscopy and fluorescence spectroscopy and microscopy. The results show that ?-spinasterol resembles the membrane behavior of cholesterol to some degree. Full article

(This article belongs to the Special Issue New Methodologies in Natural Product Analytics and Structure Elucidation)

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Figure 1
Chemical structures of (a) cholesterol and (b) α-spinasterol. Full article ">Figure 2
2H-NMR chain order parameter of the sn-1 chain of POPC-d31 in the presence of 10 and 20 mol % α-spinasterol. For comparison, the chain order parameters of pure POPC-d31 and POPC-d31 membranes in the presence of 10 and 20 mol % cholesterol are shown (data adopted from [<a href="#B18-molecules-22-01390" class="html-bibr">18</a>]). The error of measurement is smaller than the symbol size. Full article ">Figure 3
Rate constants (kp) for the permeation of dithionite across large unilamellar vesicle (LUV) membranes composed of POPC in the absence and in the presence of 20 mol % cholesterol or α-spinasterol at 37 °C. All single values of the rate constant are shown which were determined from two independent samples each threefold measured. Full article ">Figure 4
Confocal fluorescence images of Giant Unilamellar Vesicles (GUVs) containing DOPC/SSM/cholesterol (1:1:1) (a) or DOPC/SSM/α-spinasterol (1:1:1) (b). The GUV membranes were labeled with N-Rh-DOPE (0.5 mol %) that sorts preferentially into liquid disordered (ld) domains. Bar corresponds to 10 µm. Full article ">Figure 5
Distribution of the lipid analog N-Rh-DOPE (0.5 mol %) between the lo and the ld domain in GUVs consisting of DOPC/SSM/cholesterol (1:1:1) (black column) or DOPC/SSM/α-spinasterol (1:1:1) (gray column). From images as shown in <a href="#molecules-22-01390-f004" class="html-fig">Figure 4</a>, fluorescence intensities of N-Rh-DOPE localized in lo and ld domains were determined and a lo/ld ratio was calculated (see Materials and Methods). The data represent the mean ± SE of 60 vesicles, p-value < 3% (corresponding to *). Full article ">

1274 KiB

Open AccessArticle

Base-Mediated One-Pot Synthesis of Aliphatic Diazirines for Photoaffinity Labeling

by Lei Wang, Zetryana Puteri Tachrim, Natsumi Kurokawa, Fumina Ohashi, Yasuko Sakihama, Yasuyuki Hashidoko and Makoto Hashimoto

Molecules 2017, 22(8), 1389; https://doi.org/10.3390/molecules22081389 - 22 Aug 2017

Cited by 17 | Viewed by 11665

Abstract

Aliphatic diazirines have been widely used as prominent photophores for photoaffinity labeling owing to their relatively small size which can reduce the steric effect on the natural interaction between ligands and proteins. Based on our continuous efforts to develop efficient methods for the [...] Read more.

Aliphatic diazirines have been widely used as prominent photophores for photoaffinity labeling owing to their relatively small size which can reduce the steric effect on the natural interaction between ligands and proteins. Based on our continuous efforts to develop efficient methods for the synthesis of aliphatic diazirines, we present here a comprehensive study about base-mediated one-pot synthesis of aliphatic diazirines. It was found that potassium hydroxide (KOH) can also promote the construction of aliphatic diazirine with good efficiency. Importantly, KOH is cheaper, highly available, and easily handled and stored compared with the previously used base, potassium tert-butoxide (t-BuOK). Gram-scale study showed that it owned great advantages in being used for the large-scale production of aliphatic diazirines. This protocol is highly neat and the desired products can be easily isolated and purified. As the first comprehensive study of the base-mediated one-pot synthesis of aliphatic diazirines, this work provided good insight into the preparation and utilization of diazirine-based photoaffinity labeling probes. Full article

(This article belongs to the Section Medicinal Chemistry)

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9573 KiB

Open AccessArticle

Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii

by Beibei Xiang, Xiaoxue Li, Yan Wang, Xiaoxuan Tian, Zhen Yang, Lin Ma, Xia Liu and Yong Wang

Molecules 2017, 22(8), 1387; https://doi.org/10.3390/molecules22081387 - 22 Aug 2017

Cited by 17 | Viewed by 5924

Abstract

Swertia mussotii is an important medicinal plant found on the Qinghai Tibetan Plateau that has great economic and medicinal value. This plant has enjoyed a long history of use as a curative for hepatitis. The biological activity of secoiridoids, including gentiopicroside and swertiamarin, [...] Read more.

Swertia mussotii is an important medicinal plant found on the Qinghai Tibetan Plateau that has great economic and medicinal value. This plant has enjoyed a long history of use as a curative for hepatitis. The biological activity of secoiridoids, including gentiopicroside and swertiamarin, has been mainly tested for its anti-hepatitis effects. Here, we identify two candidate genes (SmIS1 and SmIS2) that are homologues of iridoid synthase and that are components of the secoiridoid pathway in S. mussotii. Using sequencing and phylogenetic analyses, we confirm that SmIS1 and SmIS2 contain six conserved short-chain dehydrogenases/reductase (SDR) motifs and thus belong to the P5?Rs group. The two purified Escherichia coli-expressed proteins reduced 8-oxogeranial to both nepetalactol and iridodials. A comparison of the kinetic parameters of SmIS1 and SmIS2 recombinant proteins revealed that SmIS2 has a lower affinity than SmIS1 for 8-oxogeranial. Transcript levels of the two genes were analysed in three different tissues of S. mussotii using semi-quantitative RT-PCR and RT-qPCR. SmIS1 and SmIS2 expression levels were more abundant in leaves and stems. This investigation adds to our knowledge of P5?Rs genes in the secoiridoid synthesis pathway and provides candidate genes for genetically improving S. mussotii by enhancing secondary metabolite production. Full article

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Figure 1
Schematic representation of the proposed in secoiridoid pathway. GPPS, geranyl diphosphate synthase; GES, geraniol synthase; G8H, geraniol 8-hydroxylase; 8-HGO, 8-hydroxygeraniol oxidoreductase; IS, iridoid synthase; IO, iridoid oxidase; 7-DLGT, 7-deoxyloganetic acid glucosyltransferase; 7-DLH, 7-deoxyloganic acid hydroxylase; LAMT, loganic acid O-methyltransferase; SLS, secologanin synthase. Genes in bold letters were previously cloned in S. mussotii; the gene in red was identified in this study. Full article ">Figure 2
Amino acid sequence comparison of SmIS1 and Smis2 with other ISs. CrIS (C. roseus, GenBank: AFW98981.1); OeIS (O. europaea, GenBank: ALV83438.1). The conserved motifs of the P5βRs are indicated with bars. The highly conserved residues are highlighted in black, whereas similar residues are highlighted in grey. Full article ">Figure 3
Predicted three-dimensional homology structure of SmIS1 (A) and SmIS2 (B) using recombinant CrIS from C. roseus (5COB) as a template. Helices and sheets are represented in red and blue, respectively. Full article ">Figure 4
Phylogenetic tree of SmIS1 and SmIS2 together with 34 known P5βRs. The phylogenetic tree was drawn using P. patens (GenBank: EDQ81106.1) as an outgroup and the following amino acid sequences with accession IDs noted in brackets: C. roseus P5bR1 (AIW09143.1); C. roseus P5bR3(AIW09145.1); Nerium oleander P5bR-B (ADG56540.1); C. roseus P5bR2 (AIW09144.1); Gomphocarpus fruticosus P5bR (ADG56546.1); Asclepias curassavica P5bR (ADG56538.1); Withania somnifera P5bR (AEY82379.1); Nicotiana tabacum P5bR (BAH47641.1); Duboisia hopwoodii P5bR (AFZ41795.1); Genlisea aurea P5bR (EPS65468.1); Mentha x piperita P5bR (ADG46022.1); Digitalis lanata P5bR1(AAS93804.1); Digitalis purpurea P5bR1(AAS93805.1); Vitis vinifera P5bR (ALB78111.1); Thlaspi densiflorum P5bR (ALD83449.1); Arabidopsis thaliana At4g24220 (NP_001078438.1); Medicago truncatula P5bR1 (AIW09149.1); Medicago truncatula P5bR2 (AIW09150.1); Passiflora incarnata P5bR (AFW16644.1); C. roseus P5bR 4 (AIW09146.1); Olea europaea 1,4-R 1.1B (ALV83440.1); Olea europaea 1,4-R 1.1A (ALV83439.1); Olea europaea 1,4-R 1.2B (ALV83442.1); Olea europaea 1,4-R 1.2A (ALV83441.1); Camptotheca acuminata CaIS (AON76722.1); Lonicera japonica LjIS (AMB61018.1); C. roseus CrIS (AFW98981.1); Olea europaea OeIS (ALV83438.1); D. purpurea P5bR2 (ACZ66261.1); D. lanata P5bR2 (ADL28122.1); C. roseus P5bR6 (AIW09148.1); Medicago truncatula P5bR3 (AIW09151.1); Medicago truncatula P5bR3 (AIW09152.1). Full article ">Figure 5
Heterologous expression of SmIS1 and Smis2 cDNA in E. coli. Lane SmIS1, purified recombinant SmIS1; lane SmIS2, purified recombinant SmIS2. Full article ">Figure 6
Functional characterization of SmIS1 and Smis2. (A) Analysis of the reaction product by GC-MS. Control, the reaction in the presence of denatured SmIS1 or SmIS2 proteins; (B) MS spectra of the substrates (peaks 1 and 2) and products (peaks 3, 4, and 5). Full article ">Figure 7
Expression pattern of the SmIS1 and SmIS2 genes in planta. (A) Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis in leaf, stems, and flower. Actin gene was used as a loading control; (B) Real-time quantitative PCR (RT-qPCR) analysis in leaf, stems, and flower. Full article ">

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Open AccessArticle

Effects of Refined Xiaoyaosan on Depressive-Like Behaviors in Rats with Chronic Unpredictable Mild Stress through Neurosteroids, Their Synthesis and Metabolic Enzymes

by Xiaoling Guo, Wenqi Qiu, Yueyun Liu, Yifang Zhang, Hongbo Zhao and Jiaxu Chen

Molecules 2017, 22(8), 1386; https://doi.org/10.3390/molecules22081386 - 21 Aug 2017

Cited by 28 | Viewed by 5305

Abstract

Abstract: To observe the effects of refined Xiaoyaosan (XYS) on the depressive-like behaviors in rats with chronic unpredictable mild stress (CUMS), and to explore the relationship between the changes of neurosteroids and mRNA expressions of their synthesis and metabolic enzymes, and [...] Read more.

Abstract: To observe the effects of refined Xiaoyaosan (XYS) on the depressive-like behaviors in rats with chronic unpredictable mild stress (CUMS), and to explore the relationship between the changes of neurosteroids and mRNA expressions of their synthesis and metabolic enzymes, and the mechanism of XYS in the treatment of depression. Methods: Eighty-four healthy male Sprague-Dawley rats were randomly divided into normal group, model group, XYS group and fluoxetine group. The latter three groups were subjected to 21 days of CUMS to prepare the stress depression model. Rats in the XYS group, and fluoxetine group were given intragastric administration with refined XYS and fluoxetine, respectively. The behavioral changes of the rats were observed after 21 days. The contents of pregnenolone (PREG), progesterone (PROG) and alloprognanolone (ALLO) in the plasma of rats were measured by ELISA. The levels of PREG, PROG and ALLO in the hippocampus and amygdala tissues were measured by LC-MS/MS. The mRNA expressions of 3?-hydroxysteroid dehydrogenase (3?-HSD), 3?-hydroxysteroid dehydrogenase (3?-HSD), cholesterol side-chain cleavage enzyme (P450scc) and 5?-reductase (5a-R) in the hippocampus and amygdala were detected by RT-qPCR methods. Results: There were changes in the model rats. The contents of PREG, PROG and ALLO changed similarly, which reflected in the decrease of PROG and ALLO, and the increase of PREG. The mRNA expression of P450scc was increased, and the mRNA expressions of 3?-HSD, 3?-HSD and 5a-R were decreased. Refined XYS could improve the behaviors of rats and the biological indicators. Conclusions: There is a neurosteroid dysfunction in the brain region of depression rat model animals, and the mechanism of refined XYS depression treatment may be related to the regulation of the control of mRNA expression of related synthesis and metabolic enzymes in the hippocampus and amygdala, further affecting the contents of neurosteroids. Full article

(This article belongs to the Special Issue Herbal Remedies Meet Modern Day Medicine: Challenges and Opportunities)

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5591 KiB

Open AccessReview

The Genus Alnus, A Comprehensive Outline of Its Chemical Constituents and Biological Activities

by Xueyang Ren, Ting He, Yanli Chang, Yicheng Zhao, Xiaoyi Chen, Shaojuan Bai, Le Wang, Meng Shen and Gaimei She

Molecules 2017, 22(8), 1383; https://doi.org/10.3390/molecules22081383 - 21 Aug 2017

Cited by 42 | Viewed by 8064

Abstract

The genus Alnus (Betulaceae) is comprised of more than 40 species. Many species of this genus have a long history of use in folk medicines. Phytochemical investigations have revealed the presence of diarylheptanoids, polyphenols, flavonoids, terpenoids, steroids and other compounds. Diarylheptanoids, natural products [...] Read more.

The genus Alnus (Betulaceae) is comprised of more than 40 species. Many species of this genus have a long history of use in folk medicines. Phytochemical investigations have revealed the presence of diarylheptanoids, polyphenols, flavonoids, terpenoids, steroids and other compounds. Diarylheptanoids, natural products with a 1,7-diphenylheptane structural skeleton, are the dominant constituents in the genus, whose anticancer effect has been brought into focus. Pure compounds and crude extracts from the genus exhibit a wide spectrum of pharmacological activities both in vitro and in vivo. This paper compiles 273 naturally occurring compounds from the genus Alnus along with their structures and pharmacological activities, as reported in 138 references. Full article

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1330 KiB

Open AccessArticle

Composition and Antioxidant, Antienzymatic and Antimicrobial Activities of Volatile Molecules from Spanish Salvia lavandulifolia (Vahl) Essential Oils

by Ana-Belen Cutillas, Alejandro Carrasco, Ramiro Martinez-Gutierrez, Virginia Tomas and Jose Tudela

Molecules 2017, 22(8), 1382; https://doi.org/10.3390/molecules22081382 - 21 Aug 2017

Cited by 27 | Viewed by 6262

Abstract

The current study describes the composition of Salvia lavandulifolia (Vahl) essential oils (SlEOs) obtained from plants cultivated in Murcia (Spain), as determined by gas chromatography. Relative and absolute concentrations, the enantiomeric ratios of chiral compounds and the in vitro antioxidant, antienzymatic and antimicrobial [...] Read more.

The current study describes the composition of Salvia lavandulifolia (Vahl) essential oils (SlEOs) obtained from plants cultivated in Murcia (Spain), as determined by gas chromatography. Relative and absolute concentrations, the enantiomeric ratios of chiral compounds and the in vitro antioxidant, antienzymatic and antimicrobial activities are described. The main components of the SlEOs were camphor, 1,8-cineole, camphene and ?-pinene, and the main enantiomers were (+)-camphor and (?)-camphene. The activities against free radicals and the capacity to reduce and chelate metallic ions were measured. SlEO-3 showed the highest activity in ORAC, DPPH, ABTS and reducing power methods, while SlEO-1 exhibited the highest chelating power. The activity of lipoxygenase and acetylcholinesterase could be inhibited by all the SlEOs, being bornyl acetate and limonene the most active individual compounds against lipoxygenase and 1,8-cineole against acetylcholinesterase. SlEOs and some individual compounds inhibited Escherichia coli, Staphylococcus aureus and Candida albicans. These results increase our knowledge of SlEOs and, particularly, provide for the first time a complete characterization of SlEOs from Murcia, Spain, while proposing possible biotechnological uses for them. Full article

(This article belongs to the Special Issue Essential Oils: Chemistry and Bioactivity)

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Graphical abstract

711 KiB

Open AccessArticle

Cryochemical Synthesis of Polymorphous Nanostructures of a Steroid Neurohormone

by Yurii Morozov, Dmitry Chistyakov, Vladimir Chernyshev and Gleb Sergeev

Molecules 2017, 22(8), 1378; https://doi.org/10.3390/molecules22081378 - 21 Aug 2017

Cited by 4 | Viewed by 3997

Abstract

A new cryochemical strategy of producing nanoparticles and polymorphous nanostructures of drugs is used, which is based on the dynamic combination of high and low temperatures, gas and solid phases, and inert carrier gases. This technology is applied to the synthesis of nanoparticles [...] Read more.

A new cryochemical strategy of producing nanoparticles and polymorphous nanostructures of drugs is used, which is based on the dynamic combination of high and low temperatures, gas and solid phases, and inert carrier gases. This technology is applied to the synthesis of nanoparticles of steroid neurohormone dehydroepiandrosterone (DHEA). We have optimized the conditions of synthesis of the new polymorphous DHEA structure, FVII. The molecules of DHEA in FVII structure are bound by hydrogen bonds via oxygen atoms. The grain size is 100 nm. It is shown that the yield and ratio of the resulting nanoforms of this hormone are determined by the nature and properties of the inert carrier gas. The highest yield and selectivity of FVII are observed when carbon dioxide is used as the carrier gas. In the case of helium, the FVII content decreases from 85 to 30% and other structures are formed. In experiments without carrier gas, nanoparticles are formed but no FVII is produced. The selectivity and the effect of carrier gas are considered on the basis of homogeneous and heterogeneous formation of nanoparticles and the relationship between particle selectivity and its activity. The synthesis of various polymorphous structures on the nanoscale is assumed to be the manifestation of the size effect in the synthesis of drugs. Full article

(This article belongs to the Section Molecular Diversity)

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3283 KiB

Open AccessArticle

Synthesis of New Hydrazone Derivatives for MAO Enzymes Inhibitory Activity

by Nafiz Öncü Can, Derya Osmaniye, Serkan Levent, Begüm Nurpelin Sağlık, Beril İnci, Sinem Ilgın, Yusuf Özkay and Zafer Asım Kaplancıklı

Molecules 2017, 22(8), 1381; https://doi.org/10.3390/molecules22081381 - 20 Aug 2017

Cited by 55 | Viewed by 8920

Abstract

In the present work, 14 new 1-substituted-2-phenylhydrazone derivatives were synthesized to evaluate their inhibitory activity against hMAO enzymes. The structures of the newly synthesized hydrazones 2a–2n were characterized by IR, 1H-NMR, 13C-NMR, HR-MS spectroscopic methods. The inhibitory activity of compounds 2a–2n against hMAO-A [...] Read more.

In the present work, 14 new 1-substituted-2-phenylhydrazone derivatives were synthesized to evaluate their inhibitory activity against hMAO enzymes. The structures of the newly synthesized hydrazones 2a–2n were characterized by IR, 1H-NMR, 13C-NMR, HR-MS spectroscopic methods. The inhibitory activity of compounds 2a–2n against hMAO-A and hMAO-B enzymes was elucidated by using an in-vitro Amplex Red® reagent assay based on fluorometric methods. According to the activity studies, 2a and 2b were found to be the most active compounds against hMAO-A enzyme, with IC50 values of 0.342 µM and 0.028 µM, respectively. The most active compounds 2a–2b were evaluated by means of enzyme kinetics and docking studies. Moreover, these compounds were subjected to cytotoxicity and genotoxicity tests to establish their preliminary toxicological profiles and were found to be non-cytotoxic and non-genotoxic. Consequently, the findings of this study display the biological importance of compounds 2a, 2b as selective, irreversible and competitive inhibitors of hMAO-A. Docking studies revealed that there is a strong interaction between hMAO-A and the most active compound 2b. Full article

(This article belongs to the Collection Molecular Docking)

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Figure 1
Structures of some MAO inhibitors and the synthesized compounds 2a, 2b. Full article ">Figure 2
(A) Lineweaver-Burk plots for the inhibition of hMAO-A by compound 2a. [S], substrate concentration (μM); V, reaction velocity (nmol/min/mg protein). Inhibitor concentrations (IC50/2, IC50, and 2 × IC50) are shown at the left along with negative control. Km values from IC50/2 to Control; 2.071, 2.899, 4.550 and 0.940 (μM). Vmax value of the competitive inhibition; 85.517 ± 2.332 (nmol/min/mg protein). (B) Secondary plot for calculation of steady-state inhibition constant (Ki) of compound 2a. Equation corresponding to the line at this graph is y = 0.0634x + 0.0119, R2 = 0.9974. Ki was calculated as 0.188 μM. Full article ">Figure 3
(A) Lineweaver-Burk plots for the inhibition of hMAO-A by compound 2b. [S], substrate concentration (μM); V, reaction velocity (nmol/min/mg protein). Inhibitor concentrations (IC50/2, IC50, and 2 × IC50) are shown at the left along with negative control. Km values from IC50/2 to control; 2.831, 3.398, 5,014 and 0.940 (μM). Vmax value of the competitive inhibition; 79.360 ± 5.704 (nmol/min/mg protein). (B) Secondary plot for calculation of steady-state inhibition constant (Ki) of compound 2b. Equation corresponding to the line at this graph is y = 1.1939x + 0.0127, R2 = 0.9915. Ki was calculated as 0.011 μM. Full article ">Figure 4
Dose-response curve of compound 2a against TA98 and TA100 in the presence and absence of S9 according to AMES MPF test. Full article ">Figure 5
Dose-response curve of compound 2b against TA98 and TA100 in the presence and absence of S9 according to AMES MPF test. Orange triangle shows t test p value (unpaired 1-sided) < 0.05 with > 2-fold induction over baseline. Full article ">Figure 6
The mode of interaction of compound 2b in the active region of hMAO-A. The inhibitor (grey colored) and the important residues in the active site of the enzyme are presented by a tube model. The FAD molecule is colored orange with a ball and stick model. Full article ">Figure 7
The mode of interaction of moclobemide in the active region of hMAO-A. The inhibitor (green colored), and the important residues in the active site of the enzyme are presented by tube models. The FAD molecule is colored orange with a ball and stick model. Full article ">Scheme 1
Synthesis of target compounds 2a–2n. Full article ">

789 KiB

Open AccessArticle

Chemical Constituents of Murraya tetramera Huang and Their Repellent Activity against Tribolium castaneum

by Chun-Xue You, Shan-Shan Guo, Wen-Juan Zhang, Zhu-Feng Geng, Jun-Yu Liang, Ning Lei, Shu-Shan Du and Zhi-Wei Deng

Molecules 2017, 22(8), 1379; https://doi.org/10.3390/molecules22081379 - 20 Aug 2017

Cited by 13 | Viewed by 5229

Abstract

Sixteen compounds were isolated from the leaves and stems of Murraya tetramera Huang. Based on the NMR and MS spectral results, the structures were determined. It was confirmed that the isolated compounds included three new compounds (9, 10 and 13) [...] Read more.

Sixteen compounds were isolated from the leaves and stems of Murraya tetramera Huang. Based on the NMR and MS spectral results, the structures were determined. It was confirmed that the isolated compounds included three new compounds (9, 10 and 13) and one new natural product (8), which were identified asmurratetra A (9), murratetra B (10), murratetra C (13) and [2-(7-methoxy-2-oxochromen-8-yl)-3-methylbut-2-enyl]3-methylbut-2-enoate (8), respectively. Meanwhile, the repellent activity against Tribolium castaneum was investigated for 13 of these isolated compounds. The results showed that the tested compounds had various levels of repellent activity against T. castaneum. Among them, compounds 1 (4(15)-eudesmene-1?,6?-diol), 11 (isoferulic acid) and 16 (2,3-dihydroxypropyl hexadecanoate) showed fair repellent activity against T. castaneum. They might be considered as potential leading compounds for the development of natural repellents. Full article

(This article belongs to the Collection Bioactive Compounds)

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1188 KiB

Open AccessArticle

Tissue-Specific Accumulation of Sulfur Compounds and Saponins in Different Parts of Garlic Cloves from Purple and White Ecotypes

by Gianfranco Diretto, Angela Rubio-Moraga, Javier Argandoña, Purificación Castillo, Lourdes Gómez-Gómez and Oussama Ahrazem

Molecules 2017, 22(8), 1359; https://doi.org/10.3390/molecules22081359 - 20 Aug 2017

Cited by 73 | Viewed by 6554

Abstract

This study set out to determine the distribution of sulfur compounds and saponin metabolites in different parts of garlic cloves. Three fractions from purple and white garlic ecotypes were obtained: the tunic (SS), internal (IS) and external (ES) parts of the clove. Liquid [...] Read more.

This study set out to determine the distribution of sulfur compounds and saponin metabolites in different parts of garlic cloves. Three fractions from purple and white garlic ecotypes were obtained: the tunic (SS), internal (IS) and external (ES) parts of the clove. Liquid Chromatography coupled to High Resolution Mass spectrometry (LC-HRMS), together with bioinformatics including Principal Component Analysis (PCA), Hierarchical Clustering (HCL) and correlation network analyses were carried out. Results showed that the distribution of these metabolites in the different parts of garlic bulbs was different for the purple and the white ecotypes, with the main difference being a slightly higher number of sulfur compounds in purple garlic. The SS fraction in purple garlic had a higher content of sulfur metabolites, while the ES in white garlic was more enriched by these compounds. The correlation network indicated that diallyl disulfide was the most relevant metabolite with regards to sulfur compound metabolism in garlic. The total number of saponins was almost 40-fold higher in purple garlic than in the white variety, with ES having the highest content. Interestingly, five saponins including desgalactotigonin-rhamnose, proto-desgalactotigonin, proto-desgalactotigonin-rhamnose, voghieroside D1, sativoside B1-rhamnose and sativoside R1 were exclusive to the purple variety. Data obtained from saponin analyses revealed a very different network between white and purple garlic, thus suggesting a very robust and tight coregulation of saponin metabolism in garlic. Findings in this study point to the possibility of using tunics from purple garlic in the food and medical industries, since it contains many functional compounds which can be exploited as ingredients. Full article

(This article belongs to the Section Molecular Diversity)

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Figure 1
Heatmap visualization of sulfur compounds (A) and saponins (B) detected in three tissues (SS, ES and IS) of the white and purple ecotypes. Data represent, for each metabolite, the fold over the internal standard (IS) intensity. Full article ">Figure 2
Hierarchical clustering (HCL) visualization, applied to rows and columns, of sulfur compounds (A) and saponins (B) detected in three tissues (SS, ES and IS) of the white and purple ecotypes of garlic. Data represent, for each metabolite, the fold over the internal standard (IS) intensity. Full article ">Figure 3
Principal component analysis (PCA), according to the metabolites, of sulfur compounds (A) and saponins; (B) detected in three tissues (SS, ES and IS) of the white and purple ecotypes of garlic. Data represent, for each metabolite, the fold over the internal standard (IS) intensity. Full article ">Figure 4
Correlation networks of sulfur compounds (A) and saponins (B) detected in three tissues (SS, ES and IS) of the white and purple ecotypes of garlic. Pearson correlation (ρ) coefficients were calculated for each metabolite pair. Node colors identify sulfur compounds (light orange, left) and saponins (light blue), while node size was proportional to node strength (ns). Red and blue edges refer to, respectively, positive and negative correlations. Edge thickness was according to the corresponding |ρ|. Only ρ >|0.50| were shown. Full article ">

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Open AccessArticle

Germination under Moderate Salinity Increases Phenolic Content and Antioxidant Activity in Rapeseed (Brassica napus var oleifera Del.) Sprouts

by Beatrice Falcinelli, Valeria Sileoni, Ombretta Marconi, Giuseppe Perretti, Muriel Quinet, Stanley Lutts and Paolo Benincasa

Molecules 2017, 22(8), 1377; https://doi.org/10.3390/molecules22081377 - 19 Aug 2017

Cited by 54 | Viewed by 6312

Abstract

The use of sprouts in the human diet is becoming more and more widespread because they are tasty and high in bioactive compounds and antioxidants, with related health benefits. In this work, we sprouted rapeseed under increasing salinity to investigate the effect on [...] Read more.

The use of sprouts in the human diet is becoming more and more widespread because they are tasty and high in bioactive compounds and antioxidants, with related health benefits. In this work, we sprouted rapeseed under increasing salinity to investigate the effect on free and bound total phenolics (TP), non-flavonoids (NF), tannins (TAN), phenolic acids (PAs), and antioxidant activity. Seeds were incubated at 0, 25, 50, 100, 200 mM NaCl until early or late sprout stage, i.e., before or after cotyledon expansion, respectively. Sprouting and increasing salinity slightly decreased the bound fractions of TP, NF, TAN, PAs, while it increased markedly the free ones and their antioxidant activity. Further increases were observed in late sprouts. Moderate salinity (25–50 mM NaCl) caused the highest relative increase in phenolic concentration while it slightly affected sprout growth. On the contrary, at higher NaCl concentrations, sprouts grew slowly (100 mM NaCl) or even died before reaching the late sprout stage (200 mM). Overall, moderate salinity was the best compromise to increase phenolic content of rapeseed sprouts. The technique may be evaluated for transfer to other species as a cheap and feasible way to increase the nutritional value of sprouts. Full article

(This article belongs to the Collection Phytochemicals: Biosynthesis, Metabolism and Biological Activities)

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Open AccessArticle

Impact of Xanthylium Derivatives on the Color of White Wine

by Franziska Bührle, Anita Gohl and Fabian Weber

Molecules 2017, 22(8), 1376; https://doi.org/10.3390/molecules22081376 - 19 Aug 2017

Cited by 17 | Viewed by 6889

Abstract

Xanthylium derivatives are yellow to orange pigments with a glyoxylic acid bridge formed by dimerization of flavanols, which are built by oxidative cleavage of tartaric acid. Although their structure and formation under wine-like conditions are well established, knowledge about their color properties and [...] Read more.

Xanthylium derivatives are yellow to orange pigments with a glyoxylic acid bridge formed by dimerization of flavanols, which are built by oxidative cleavage of tartaric acid. Although their structure and formation under wine-like conditions are well established, knowledge about their color properties and their occurrence and importance in wine is deficient. Xanthylium cations and their corresponding esters were synthesized in a model wine solution and isolated via high-performance countercurrent chromatography (HPCCC) and solid phase extraction (SPE). A Three-Alternative-Forced-Choice (3-AFC) test was applied to reveal the color perception threshold of the isolated compounds in white wine. Their presence and color impact was assessed in 70 different wines (58 white and 12 rosé wines) by UHPLC-DAD-ESI-MSⁿ and the storage stability in wine was determined. The thresholds in young Riesling wine were 0.57 mg/L (cations), 1.04 mg/L (esters) and 0.67 mg/L (1:1 (w/w) mixture), respectively. The low thresholds suggest a possible impact on white wine color, but concentrations in wines were below the threshold. The stability study showed the degradation of the compounds during storage under several conditions. Despite the low perception threshold, xanthylium derivatives might have no direct impact on white wine color, but might play a role in color formation as intermediate products in polymerization and browning. Full article

(This article belongs to the Collection Wine Chemistry)

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Mechanism of the xanthylium cation (NJ2) formation and its corresponding ethyl ester (NJ3) from (+)-catechin and glyoxylic acid via a colorless carboxymethine-linked dimer (dimer 2a) and colorless xanthene [<a href="#B18-molecules-22-01376" class="html-bibr">18</a>]. Full article ">Figure 2
Formation of the three different carboxymethine-linked (+)-catechin dimers from (+)-catechin substituted with glyoxylic acid at ring position 6 or 8 [<a href="#B10-molecules-22-01376" class="html-bibr">10</a>,<a href="#B17-molecules-22-01376" class="html-bibr">17</a>,<a href="#B18-molecules-22-01376" class="html-bibr">18</a>]. Full article ">Figure 3
UHPLC-DAD-ESI+-MS chromatogram of the synthesis medium after an incubation of 15 days at 45 °C for the identified compounds (a) (+)-catechin (peak 2), carboxymethine-linked (+)-catechin dimers (peak 1, 3 and 4), xanthylium lactone (peak 9) at 280 nm; (b) non-esterified xanthylium cations at m/z 617 (peak 5−8, 10); (c) xanthylium cation ethyl esters at m/z 645 (peak 11−14). 1. Peaks 3 and 4 were not detectable at UV-spectrum. Full article ">Figure 4
Color parameter C* of Riesling wine and spiked wines. xc = non-esterified xanthylium cations, xce = xanthylium cation ethyl esters, xc + xce = equivalent mixture of xanthylium cations, and its corresponding ethyl esters (1:1, w/w). Bars with different letters are significantly different at p ≤ 0.05 (n = 2; mean standard deviation 3%). Full article ">Figure 5
Correlation between added amount of xanthylium derivatives and correct answers in a 3-AFC test, comparing non-esterified xanthylium cations (xc), xanthylium cation ethyl esters (xce) and the mixture of both compound classes (xc + xce). The arrows indicate the respective perception threshold. Full article ">Figure 6
UHPLC-ESI+-MS (SRM) chromatogram of the synthesis medium with catechin and epicatechin after incubation at 45 °C for 6 days. (a) Single reaction monitoring for xanthylium cations m/z 617 → 465; (b) Single reaction monitoring for xanthylium cation ethyl esters m/z 645 → 493. Peak labels indicate retention time. Full article ">Figure 7
Development of the concentration in white wine under three different storage conditions over a period of 14 days (n = 2), quantified at 440 nm as NJ2 equivalents (a) non-esterified xanthylium cations; (b) xanthylium cation ethyl esters. Full article ">

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Open AccessArticle

Pharmacological and Toxicological Screening of Novel Benzimidazole-Morpholine Derivatives as Dual-Acting Inhibitors

by Nafiz Öncü Can, Ulviye Acar Çevik, Begüm Nurpelin Sağlık, Yusuf Özkay, Özlem Atlı, Merve Baysal, Ümide Demir Özkay and Özgür Devrim Can

Molecules 2017, 22(8), 1374; https://doi.org/10.3390/molecules22081374 - 19 Aug 2017

Cited by 22 | Viewed by 6613

Abstract

The aim of this study was to investigate acetylcholinesterase (AChE), monoamine oxidase A (MAO-A), monoamine oxidase B (MAO-B), cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzyme inhibitory, and antimicrobial activities of a new series of 2-(4-substituted phenyl)-1-[2-(morpholin-4-yl)ethyl]-1H-benzimidazole derivatives, for their possible use as [...] Read more.

The aim of this study was to investigate acetylcholinesterase (AChE), monoamine oxidase A (MAO-A), monoamine oxidase B (MAO-B), cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzyme inhibitory, and antimicrobial activities of a new series of 2-(4-substituted phenyl)-1-[2-(morpholin-4-yl)ethyl]-1H-benzimidazole derivatives, for their possible use as multi-action therapeutic agents. Target compounds (n = 15) were synthesized under microwave irradiation conditions in two steps, and their structures were elucidated by FT-IR, ¹H-NMR, ¹³C-NMR and high resolution mass spectroscopic analyses. Pharmacological screening studies revealed that two of the compounds (2b and 2j) have inhibitory potential on both COX-1 and COX-2 enzymes. In addition, cytotoxic and genotoxic properties of the compounds 2b, 2j and 2m were investigated via the well-known MTT and Ames tests, which revealed that the mentioned compounds are non-cytotoxic and non-genotoxic. As a concise conclusion, two novel compounds were characterized as potential candidates for treatment of frequently encountered inflammatory diseases. Full article

(This article belongs to the Section Medicinal Chemistry)

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Open AccessArticle

Inhibitory Effect of Bovine Lactoferrin on Catechol-O-Methyltransferase

by Masayuki Ikeda, Hiroshi Iijima, Ichizo Shinoda, Hiroshi Iwamoto and Yasuhiro Takeda

Molecules 2017, 22(8), 1373; https://doi.org/10.3390/molecules22081373 - 19 Aug 2017

Cited by 8 | Viewed by 7636

Abstract

Lactoferrin (LF) is a well-known multifunctional protein. In this study, we report the inhibitory potency of bovine LF (bLF) on catechol-O-methyltransferase (COMT), which catalyzes methylation of catechol substrates. We found that bLF binds to and inhibits COMT using its N-terminal [...] Read more.

Lactoferrin (LF) is a well-known multifunctional protein. In this study, we report the inhibitory potency of bovine LF (bLF) on catechol-O-methyltransferase (COMT), which catalyzes methylation of catechol substrates. We found that bLF binds to and inhibits COMT using its N-terminal region. An N-terminal peptide fragment obtained from bLF by trypsin digestion showed a higher inhibitory activity than intact bLF. A synthetic fragment of the bLF N-terminal residues 6–50, with two pairs of disulfide bonds, also showed higher inhibitory activity than intact bLF. Enzyme kinetic studies proved that bLF did not compete with S-adenosylmethionine (the methyl donor substrate) as well as methyl acceptor substrates such as dihydroxybenzoic acid, (?)-epicatechin, norepinephrine, or l-3,4-dihydroxyphenylalanine. The inhibitory potency of bLF decreased against a COMT preparation pretreated with dithiothreitol, suggesting that the oxidation status of COMT is relevant to interaction with bLF. We further confirmed that COMT activity in the cell extracts form Caco-2 and HepG2 cells was inhibited by bLF and by the synthesized fragment. Enzyme kinetic study indicated that bLF functions as a non-competitive inhibitor by binding to an allosteric surface of COMT. Full article

(This article belongs to the Section Medicinal Chemistry)

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Full article ">Figure 1
Inhibitory activity of bovine lactoferrin (bLF)-derived peptides on catechol-O-methyltransferase (COMT): (A) The horizontal axis represents peptide concentration. The vertical axis represents relative COMT activity in %. (B) The mapping of peptides. P36, P20, and P51 are trypsin-digested fragments derived from bLF. Their N-terminal amino acid sequences were analyzed and their molecular weights were estimated by SDS-PAGE analysis. The sequence and molecular weights determined their mapping as presented in this panel. (C) Amino acid sequence of LFcinB and P5. Basic amino acids (K, R) are shown in blue, and acidic amino acids (E) are shown in red. The two disulfide bonds are conserved in native bLF. (D) Stereo view (wall-eye) representation of bLF (Protein Data Bank entry 1BLF [<a href="#B39-molecules-22-01373" class="html-bibr">39</a>]). P5 (6–50) region: red; LFcinB (16–41) region: green; C-lobe (P51 285–689) region: light brown. Disulfide bridges (Cys9–Cys45 and Cys19–Cys36) are shown. Full article ">Figure 2
Enzyme kinetic analysis of the inhibitory activity of bLF on COMT activity: (A) Lineweaver–Burk plot of the COMT reaction with varying concentrations of S-adenosylmethionine (SAM) (4, 11.2, and 83.2 μM). KmSAM was calculated from the intercepts of the horizontal axis to be 11.4, 8.0, and 10.3 μM. (B) Lineweaver–Burk plot of the COMT reaction with varying concentrations of dihydroxybenzoic acid (DBA) (2, 5, 20, 50, 200, and 500 μM). The concentrations of bLF in the reaction mixture are given in the graph legend. Full article ">Figure 3
Effects of bLF and P5 on dithiothreitol DTT)-treated COMT (time course experiments): COMT was treated with 1.2 mM DTT for 0–60 min at 37 °C, and then the catalytic activity of the treated preparations was measured in the presence or absence of bLF or P5 (DTT final concentration: 1.0 mM). The horizontal axis represents the duration of DTT treatment of COMT in minutes. The vertical axis represents the catalytic activity of COMT expressed by the amount of methyl transfer (scintillation counts). This experiment was performed with triplicate measurements and values were described as average and standard deviation (SD). Full article ">Figure 4
Effects of bLF and P5 on COMT activity in cell lysates: (A) Immunostaining of COMT (lanes 2–5) and Coomassie staining (lanes 6–9) of protein extracted from HepG2 and Caco-2 cells. Lanes 2–9 were loaded with equal volumes of cell lysates. Loaded proteins: 15 μg (HepG2), 20 μg (Caco-2 −DTT), and 22 μg (Caco-2 +DTT). Lane 1 is recombinant human soluble COMT. Lysates designated as +DTT are lysates prepared using extraction buffer containing 5 mM DTT. Those designated as −DTT are lysates prepared in the absence of DTT; (B) Susceptibility of −DTT lysates to inhibitors. Effect of bLF and entacapone to COMT activity (−DTT lysates) was investigated in the reaction mixtures that contained 1 mM DTT or did not. Concentrations of bLF and entacapone were 63 μM and 0.19 μM, respectively. The vertical axis represents COMT activity expressed as counts per minute; (C) Concentration dependency of COMT activity on inhibitors. The horizontal axis represents the concentration of inhibitors. COMT activity of the −DTT lysate prepared from Caco-2 cell was examined. The reaction was performed in the absence of DTT. Full article ">Figure 5
Summary of bLF inhibitory activity on COMT. Full article ">

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Open AccessArticle

Synthesis and Antimicrobial Studies of New Antibacterial Azo-Compounds Active against Staphylococcus aureus and Listeria monocytogenes

by Stefano Piotto, Simona Concilio, Lucia Sessa, Rosita Diana, Gabriel Torrens, Carlos Juan, Ugo Caruso and Pio Iannelli

Molecules 2017, 22(8), 1372; https://doi.org/10.3390/molecules22081372 - 19 Aug 2017

Cited by 51 | Viewed by 7329

Abstract

Some novel (phenyl-diazenyl)phenols (4a–m) were designed and synthesized to be evaluated for their antibacterial activity. Starting from an active previously-synthesized azobenzene chosen as lead compound, we introduced some modifications and optimization of the structure, in order to improve solubility [...] Read more.

Some novel (phenyl-diazenyl)phenols (4a–m) were designed and synthesized to be evaluated for their antibacterial activity. Starting from an active previously-synthesized azobenzene chosen as lead compound, we introduced some modifications and optimization of the structure, in order to improve solubility and drug conveyance. Structures of all newly-synthesized compounds were confirmed by ¹H nuclear magnetic resonance (NMR), mass spectrometry, and UV-Vis spectroscopy. Antibacterial activity of the new compounds was tested with the dilution method against the bacteria strains Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa PAO1. All the compounds were selectively active against Gram-positive bacteria. In particular, compounds 4d, 4h, and 4i showed the highest activity against S. aureus and Listeria monocytogenes, reaching remarkable MIC₁₀₀ values of 4 ?g/mL and 8 ?g/mL. The relationship between antimicrobial activity and compound structure has suggested that the presence of hydroxyl groups seems to be essential for antimicrobial activity of phenolic compounds. Full article

(This article belongs to the Special Issue Antibacterial Materials and Coatings)

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Open AccessFeature PaperArticle

BET & ELF Quantum Topological Analysis of Neutral 2-Aza-Cope Rearrangement of ?-Alkenyl Nitrones

by Pedro Merino, Maria A. Chiacchio, Laura Legnani and Tomás Tejero

Molecules 2017, 22(8), 1371; https://doi.org/10.3390/molecules22081371 - 19 Aug 2017

Cited by 5 | Viewed by 5915

Abstract

The 2-Aza-Cope rearrangement of ?-alkenyl nitrones is a rare example of the neutral thermal 2-aza-Cope process that usually takes place with cationic species. During the rearrangement, a redistribution of bonds and electronic density occurs in one kinetic step. However, the introduction of substituents [...] Read more.

The 2-Aza-Cope rearrangement of ?-alkenyl nitrones is a rare example of the neutral thermal 2-aza-Cope process that usually takes place with cationic species. During the rearrangement, a redistribution of bonds and electronic density occurs in one kinetic step. However, the introduction of substituents with different steric requirements and electronic features might alter the activation energies and the synchronicity of the reaction. The electron localization function (ELF) analysis and its application to Bonding Evolution Theory (BET) analysis within the context of Molecular Electron Density Theory (MEDT) is an excellent tool to monitor the electron density along the reaction coordinate and thus investigate in detail bond breaking and formation and the corresponding energy barriers. By analyzing topological ELF calculations of seventeen 2-aza-Cope nitrone rearrangements with selected substituents, the main factors influencing the synchronicity of the process were investigated. This MEDT study results revealed that the rearrangement is a non-polar process mostly influenced by steric factors rather than by electronic ones, and confirms the pseudoradical character of the process rather than any pericyclic electron-reorganization. Full article

(This article belongs to the Special Issue The Molecular Electron Density Theory: A Modern View of Molecular Reactivity in Organic Chemistry)

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Open AccessReview

Metal-Based Nanoparticles for the Treatment of Infectious Diseases

by Blessing Atim Aderibigbe

Molecules 2017, 22(8), 1370; https://doi.org/10.3390/molecules22081370 - 18 Aug 2017

Cited by 214 | Viewed by 15167

Abstract

Infectious diseases can be transmitted and they cause a significant burden on public health globally. They are the greatest world killers and it is estimated that they are responsible for the demise of over 17 million people annually. The impact of these diseases [...] Read more.

Infectious diseases can be transmitted and they cause a significant burden on public health globally. They are the greatest world killers and it is estimated that they are responsible for the demise of over 17 million people annually. The impact of these diseases is greater in the developing countries. People with compromised immune systems and children are the most affected. Infectious diseases may be caused by bacteria, viruses, and protozoa. The treatment of infectious diseases is hampered by simultaneous resistance to multiple drugs, indicating that there is a serious and pressing need to develop new therapeutics that can overcome drug resistance. This review will focus on the recent reports of metal-based nanoparticles that are potential therapeutics for the treatment of infectious diseases and their biological efficacy (in vitro and in vivo). Full article

(This article belongs to the Special Issue Metal Based Drugs: Opportunities and Challenges)

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Open AccessReview

Anesthetic Agents of Plant Origin: A Review of Phytochemicals with Anesthetic Activity

by Hironori Tsuchiya

Molecules 2017, 22(8), 1369; https://doi.org/10.3390/molecules22081369 - 18 Aug 2017

Cited by 91 | Viewed by 16450

Abstract

The majority of currently used anesthetic agents are derived from or associated with natural products, especially plants, as evidenced by cocaine that was isolated from coca (Erythroxylum coca, Erythroxylaceae) and became a prototype of modern local anesthetics and by thymol and [...] Read more.

The majority of currently used anesthetic agents are derived from or associated with natural products, especially plants, as evidenced by cocaine that was isolated from coca (Erythroxylum coca, Erythroxylaceae) and became a prototype of modern local anesthetics and by thymol and eugenol contained in thyme (Thymus vulgaris, Lamiaceae) and clove (Syzygium aromaticum, Myrtaceae), respectively, both of which are structurally and mechanistically similar to intravenous phenolic anesthetics. This paper reviews different classes of phytochemicals with the anesthetic activity and their characteristic molecular structures that could be lead compounds for anesthetics and anesthesia-related drugs. Phytochemicals in research papers published between 1996 and 2016 were retrieved from the point of view of well-known modes of anesthetic action, that is, the mechanistic interactions with Na⁺ channels, ?-aminobutyric acid type A receptors, N-methyl-d-aspartate receptors and lipid membranes. The searched phytochemicals include terpenoids, alkaloids and flavonoids because they have been frequently reported to possess local anesthetic, general anesthetic, antinociceptive, analgesic or sedative property. Clinical applicability of phytochemicals to local and general anesthesia is discussed by referring to animal in vivo experiments and human pre-clinical trials. This review will give structural suggestions for novel anesthetic agents of plant origin. Full article

(This article belongs to the Special Issue Polypharmacology and Multitarget Drug Discovery)

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Open AccessArticle

Phase Behaviour and Miscibility Studies of Collagen/Silk Fibroin Macromolecular System in Dilute Solutions and Solid State

by Ima Ghaeli, Mariana A. De Moraes, Marisa M. Beppu, Katarzyna Lewandowska, Alina Sionkowska, Frederico Ferreira-da-Silva, Maria P. Ferraz and Fernando J. Monteiro

Molecules 2017, 22(8), 1368; https://doi.org/10.3390/molecules22081368 - 18 Aug 2017

Cited by 26 | Viewed by 6863

Abstract

Miscibility is an important issue in biopolymer blends for analysis of the behavior of polymer pairs through the detection of phase separation and improvement of the mechanical and physical properties of the blend. This study presents the formulation of a stable and one-phase [...] Read more.

Miscibility is an important issue in biopolymer blends for analysis of the behavior of polymer pairs through the detection of phase separation and improvement of the mechanical and physical properties of the blend. This study presents the formulation of a stable and one-phase mixture of collagen and regenerated silk fibroin (RSF), with the highest miscibility ratio between these two macromolecules, through inducing electrostatic interactions, using salt ions. For this aim, a ternary phase diagram was experimentally built for the mixtures, based on observations of phase behavior of blend solutions with various ratios. The miscibility behavior of the blend solutions in the miscible zones of the phase diagram was confirmed quantitatively by viscosimetric measurements. Assessing the effects of biopolymer mixing ratio and salt ions, before and after dialysis of blend solutions, revealed the importance of ion-specific interactions in the formation of coacervate-based materials containing collagen and RSF blends that can be used in pharmaceutical, drug delivery, and biomedical applications. Moreover, the conformational change of silk fibroin from random coil to beta sheet, in solution and in the final solid films, was detected by circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR), respectively. Scanning electron microscopy (SEM) exhibited alterations of surface morphology for the biocomposite films with different ratios. Surface contact angle measurement illustrated different hydrophobic properties for the blended film surfaces. Differential scanning calorimetry (DSC) showed that the formation of the beta sheet structure of silk fibroin enhances the thermal stability of the final blend films. Therefore, the novel method presented in this study resulted in the formation of biocomposite films whose physico-chemical properties can be tuned by silk fibroin conformational changes by applying different component mixing ratios. Full article

(This article belongs to the Special Issue Natural Polymers and Biopolymers)

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Open AccessFeature PaperArticle

Copper-Catalyzed Synthesis of Unsymmetrical Diorganyl Chalcogenides (Te/Se/S) from Boronic Acids under Solvent-Free Conditions

by Sumbal Saba, Giancarlo Vaccari Botteselle, Marcelo Godoi, Tiago Elias Allievi Frizon, Fábio Zazyki Galetto, Jamal Rafique and Antonio L. Braga

Molecules 2017, 22(8), 1367; https://doi.org/10.3390/molecules22081367 - 18 Aug 2017

Cited by 46 | Viewed by 7616

Abstract

The efficient and mild copper-catalyzed synthesis of unsymmetrical diorganyl chalcogenides under ligand- and solvent-free conditions is described. The cross-coupling reaction was performed using aryl boric acids and 0.5 equiv. of diorganyl dichalcogenides (Te/Se/S) in the presence of 3 mol % of CuI and [...] Read more.

The efficient and mild copper-catalyzed synthesis of unsymmetrical diorganyl chalcogenides under ligand- and solvent-free conditions is described. The cross-coupling reaction was performed using aryl boric acids and 0.5 equiv. of diorganyl dichalcogenides (Te/Se/S) in the presence of 3 mol % of CuI and 3 equiv. of DMSO, under microwave irradiation. This new protocol allowed the preparation of several unsymmetrical diorganyl chalcogenides in good to excellent yields. Full article

(This article belongs to the Special Issue Celebrating Two Centuries of Research in Selenium Chemistry: State of the Art and New Prospectives)

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Full article ">Scheme 1
Solvent- and ligand-free synthesis of unsymmetrical diorganyl chalcogenides catalyzed by CuI. Full article ">Scheme 2
Synthesis of unsymmetrical organotellurides a,b. a Reaction conditions: 1 (0.25 mmol), 2 (0.5 mmol) in the presence of CuI (3.0 mol %) and DMSO (3.0 equiv.) applied for 15 min at 100 °C with 100 W of MW-irradiation; b Isolated yields. Full article ">Scheme 3
Synthesis of unsymetrical organoselenides and sulfides a,b. a Reaction conditions: 4 or 5 (0.25 mmol), 2 (0.5 mmol) in the presence of CuI (3.0 mol %) and DMSO (3.0 equiv.) for 15 min at 100 °C and 100 W of MW-irradiations; b Isolated yields. Full article ">Scheme 4
Scale-Up of the reaction. Full article ">Scheme 5
Control experiments for reaction mechanism. (a) Reaction in the presence of radical inhibitor; (b) Reaction under inert atmosphere; (c) Reaction under oxygen atmosphere. Full article ">Scheme 6
A plausible reaction pathway. Full article ">

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Open AccessArticle

Detection of Interactions between Proteins by Using Legendre Moments Descriptor to Extract Discriminatory Information Embedded in PSSM

by Yan-Bin Wang, Zhu-Hong You, Li-Ping Li, Yu-An Huang and Hai-Cheng Yi

Molecules 2017, 22(8), 1366; https://doi.org/10.3390/molecules22081366 - 18 Aug 2017

Cited by 28 | Viewed by 5037

Abstract

Protein-protein interactions (PPIs) play a very large part in most cellular processes. Although a great deal of research has been devoted to detecting PPIs through high-throughput technologies, these methods are clearly expensive and cumbersome. Compared with the traditional experimental methods, computational methods have [...] Read more.

Protein-protein interactions (PPIs) play a very large part in most cellular processes. Although a great deal of research has been devoted to detecting PPIs through high-throughput technologies, these methods are clearly expensive and cumbersome. Compared with the traditional experimental methods, computational methods have attracted much attention because of their good performance in detecting PPIs. In our work, a novel computational method named as PCVM-LM is proposed which combines the probabilistic classification vector machine (PCVM) model and Legendre moments (LMs) to predict PPIs from amino acid sequences. The improvement mainly comes from using the LMs to extract discriminatory information embedded in the position-specific scoring matrix (PSSM) combined with the PCVM classifier to implement prediction. The proposed method was evaluated on Yeast and Helicobacter pylori datasets with five-fold cross-validation experiments. The experimental results show that the proposed method achieves high average accuracies of 96.37% and 93.48%, respectively, which are much better than other well-known methods. To further evaluate the proposed method, we also compared the proposed method with the state-of-the-art support vector machine (SVM) classifier and other existing methods on the same datasets. The comparison results clearly show that our method is better than the SVM-based method and other existing methods. The promising experimental results show the reliability and effectiveness of the proposed method, which can be a useful decision support tool for protein research. Full article

(This article belongs to the Special Issue Computational Analysis for Protein Structure and Interaction)

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Open AccessArticle

Biosynthesis of S-Adenosylmethionine by Magnetically Immobilized Escherichia coli Cells Highly Expressing a Methionine Adenosyltransferase Variant

by Chunli Yin, Tao Zheng and Xin Chang

Molecules 2017, 22(8), 1365; https://doi.org/10.3390/molecules22081365 - 18 Aug 2017

Cited by 15 | Viewed by 5609

Abstract

S-Adenosylmethionine (SAM) is a natural metabolite having important uses in the treatment of various diseases. To develop a simple and effective way to produce SAM, immobilized Escherichia coli cells highly expressing an engineered variant of methionine adenosyltransferase (MAT) were employed to synthesize SAM. [...] Read more.

S-Adenosylmethionine (SAM) is a natural metabolite having important uses in the treatment of various diseases. To develop a simple and effective way to produce SAM, immobilized Escherichia coli cells highly expressing an engineered variant of methionine adenosyltransferase (MAT) were employed to synthesize SAM. The recombinant I303V MAT variant was successfully produced at approximately 900 mg/L in a 10-L bioreactor and exhibited significantly less product inhibition and had a four-fold higher specific activity (14.2 U/mg) than the wild-type MAT (3.6 U/mg). To reduce the mass transfer resistance, the free whole-cells were permeabilized and immobilized using gellan gum gel as support in the presence of 100 mg/L Fe₃O₄ nanoparticles, and the highest activity (4152.4 U/L support) was obtained, with 78.2% of the activity recovery. The immobilized cells were more stable than the free cells under non-reactive conditions, with a half-life of 9.1 h at 50 °C. Furthermore, the magnetically immobilized cells were employed to produce SAM at a 40-mM scale. The residual activity of the immobilized cells was 67% of its initial activity after 10 reuses, and the conversion rate of ATP was ?95% in all 10 batches. These results indicated that magnetically immobilized cells should be a promising biocatalyst for the biosynthesis of SAM. Full article

(This article belongs to the Section Green Chemistry)

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Figure 1
The 3D structure of the methionine adenosyltransferase (MAT) derived from E. coli (PDB: 1RG9). (a) The dimeric MAT with S-adenosylmethionine (SAM) bound between two monomer units; (b) The binding site of the product SAM (purple carbon atoms) in the active site of MAT. The interaction between the methyl group of SAM and the isoleucine I303 was hypothesized to affect the dissociation of SAM in the active center of the enzyme. Thus, isoleucine I303 became the target of site-directed mutagenesis. Full article ">Figure 2
Conversion rates of adenosine triphosphate (ATP) catalyzed by the purified wild-type MAT and its variant I303V MAT. (a) SDS-PAGE (12%) analysis of the purified wild-type and I303V MAT proteins. Lane M: protein molecular weight marker. Lane 1: The purified wild-type MAT. Lane 2: The purified I303V MAT; (b) Production of SAM catalyzed by the wild-type and I303V MAT proteins as a function of the substrate concentration. The reaction mixtures, which contained 10 mM ATP and varying concentrations of l-methionine (0.1–10 mM), were incubated for 5 h at 37 °C in the presence of excess enzyme. The reaction yield was determined by measuring the phosphate that was released from the substrate ATP during the reaction. Curves were fitted by using the logistic model; (c,d) Effect of varying concentrations of sodium p-toluenesulfonate on the ATP conversion rate using wild-type MAT (c) and I303V MAT (d) in the presence of a 1.3-fold molar excess of l-methionine after a 6-h incubation at 37 °C, pH 7.0. The consumption of ATP and the production of SAM were determined by HPLC analysis. Full article ">Figure 3
Expression of recombinant I303V MAT in a 10-L bioreactor. (a) Effect of induction time on cell biomass (g wet cell weight/L) and the activity of recombinant I303V MAT. The recombinant E. coli cells were induced by the addition of IPTG (isopropyl β-d-1-thiogalactopyranoside) to a final concentration of 0.1 mM; (b) SDS-PAGE (10%) analysis of the expression of the recombinant I303V MAT. Lane M: Molecular weight marker. Lane 1: Total protein of recombinant E. coli before IPTG induction. Lanes 2–5: Total protein of recombinant E. coli induced with IPTG after 5, 9, 13, and 17 h, respectively. Full article ">Figure 4
Permeabilization of free whole-cells of E. coli and synthesis of SAM catalyzed by the untreated and permeabilized cells. (a) Cell-bound I303V MAT activity for E. coli cells permeabilized with different detergents (0.1% w/v) or organic solvents. Treatment was carried out for 30 min with 10 mL/g wet cell weight; (b) Conversion rate of the ATP substrate catalyzed by untreated and permeabilized E. coli cells as a function of time. Reactions were performed in a 100-mL volume and included: 40 mM ATP, 50 mM l-methionine (l-Met), 50 mM K2SO4, 100 mM MgSO4, 0.3 M sodium p-toluenesulfonate, 100 mM Tris-HCl, and 1 g of cells at 37 °C, pH 7.0. The consumption of ATP and the production of SAM were determined by HPLC analysis, and the MAT activity was calculated according to the production of SAM. Full article ">Figure 5
Effect of the concentration of Fe3O4 nanoparticles on the activity of immobilized cells, and the thermal stability of free and magnetically immobilized cells; (a) Biosynthesis of SAM by magnetically immobilized cells prepared by the addition of different concentration of Fe3O4 nanoparticles (50–150 mg/L), nonmagnetically immobilized cells, and free cells. The reaction mixtures contained: 40 mM ATP, 50 mM l-methionine, 50 mM K2SO4, 100 mM MgSO4, 0.3 M sodium p-toluenesulfonate, 100 mM Tris-HCl (pH 7.0), and the equivalent amount of biocatalyst for free or immobilized cells, at 37 °C. The conversion rate of ATP was determined by HPLC analysis; (b,c) The thermal stability of free and magnetically immobilized cells. Percent enzymatic activity remaining as a function of time for free (b); and magnetically immobilized cells (c). Assay conditions: 50 mM Tris-HCl buffer, pH 7.5. Initial activities were defined as 100%. The residual activity of free and immobilized cells was measured using the spectrophotometric assay. Enzyme activity was calculated according to the phosphate concentration that was released from the substrate ATP during the reaction. Full article ">Figure 6
Reuse of magnetically immobilized cells for SAM biosynthesis. (a) The effect of recycling on the activity of magnetically immobilized cells. The residual activity was measured using the spectrophotometric assay. Enzyme activity was calculated according to the phosphate concentration that was released from the substrate ATP during the reaction; (b) Production of SAM with magnetically immobilized cells in 10 repeated batches. The reaction was performed in 100 mM Tris-HCl buffer containing 40 mM ATP, 50 mM l-methionine, 50 mM K2SO4, 100 mM MgSO4, and 0.3 M sodium p-toluenesulfonate, pH 7.0 with a 6-h incubation at 37 °C. The consumption of ATP and the production of SAM were determined by HPLC analysis, and the conversion rate of ATP was calculated. Full article ">

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Open AccessArticle

Phytochemicals of Euphorbia lathyris L. and Their Antioxidant Activities

by Lizhen Zhang, Chu Wang, Qiuxia Meng, Qin Tian, Yu Niu and Wei Niu

Molecules 2017, 22(8), 1335; https://doi.org/10.3390/molecules22081335 - 18 Aug 2017

Cited by 21 | Viewed by 5482

Abstract

The objectives of this study were to characterize the antioxidant capacities and phytochemicals such as phenolics and flavonoids in four parts of Euphorbia lathyris L. HPLC was employed to detect the type and content of phenolic acids and flavonoids in the root, stem, [...] Read more.

The objectives of this study were to characterize the antioxidant capacities and phytochemicals such as phenolics and flavonoids in four parts of Euphorbia lathyris L. HPLC was employed to detect the type and content of phenolic acids and flavonoids in the root, stem, seed, and testa of the plant. The total phenolic content (TPC) and total flavonoid content (TFC) were different among various parts of E. lathyris. The highest TPC were found in the testa (290.46 ± 15.09 mg of gallic acid equiv/100 g dry weight (DW)). However, the root contained the highest TFC (215.68 ± 3.10 mg of rutin equiv/g DW). Of the different antioxidant activities detected, DPPH free radical scavenging activity was highest in the testa (61.29 ± 0.29 mmol Trolox/100 g DW), but the highest FRAP antioxidant activity was found in the seed (1131.25 ± 58.68 mg FeSO₄/100 g DW of free compounds and 1927.43 ± 52.13 mg FeSO₄/100 g DW of bound compounds). There was a positive correlation between the total phenolic contents and DPPH free radical scavenging activity in different parts of E. lathyris. Full article

(This article belongs to the Special Issue Selected Papers from the 6th International Conference on Biotechnology and Bioengineering (6-ICBB 2017))

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Open AccessReview

Significance of Resveratrol in Clinical Management of Chronic Diseases

by Awais Wahab, Kuo Gao, Caixia Jia, Feilong Zhang, Guihua Tian, Ghulam Murtaza and Jianxin Chen

Molecules 2017, 22(8), 1329; https://doi.org/10.3390/molecules22081329 - 18 Aug 2017

Cited by 92 | Viewed by 9072

Abstract

Resveratrol could be beneficial to health and provides protection against a wide array of pathologies and age-associated problems, as evident from preclinical studies. However, a comparison of animal and human studies reveals that this dietary polyphenol cannot protect against metabolic diseases and their [...] Read more.

Resveratrol could be beneficial to health and provides protection against a wide array of pathologies and age-associated problems, as evident from preclinical studies. However, a comparison of animal and human studies reveals that this dietary polyphenol cannot protect against metabolic diseases and their associated complications. The clinical outcomes are affected by many factors such as sample size. This article not only presents a comprehensive review of the current advances concerning the dose, the extent of absorption, interaction and toxicity of resveratrol in human studies, but also describes its therapeutic effects against several chronic diseases such as diabetes mellitus, obesity, cardiovascular diseases, cancer and aging and the related diseases. Full article

(This article belongs to the Special Issue Improvements for Resveratrol Efficacy)

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17205 KiB

Open AccessArticle

Biochemical and Comparative Transcriptomic Analyses Identify Candidate Genes Related to Variegation Formation in Paeonia rockii

by Qianqian Shi, Long Li, Xiaoxiao Zhang, Jianrang Luo, Xiang Li, Lijuan Zhai, Lixia He and Yanlong Zhang

Molecules 2017, 22(8), 1364; https://doi.org/10.3390/molecules22081364 - 17 Aug 2017

Cited by 23 | Viewed by 5596

Abstract

Paeonia rockii is a wild tree peony species with large and dark purple variegations at the base of its petals. It is the genetic resource for various variegation patterns in tree peony cultivars, which is in contrast to the pure white petals of [...] Read more.

Paeonia rockii is a wild tree peony species with large and dark purple variegations at the base of its petals. It is the genetic resource for various variegation patterns in tree peony cultivars, which is in contrast to the pure white petals of Paeonia ostii. However, the molecular mechanism underlying the formation of variegation in this plant is still unknown. Here, we conducted Illumina transcriptome sequencing for P. rockii, P. ostii (with pure white petals) and their F1 individuals (with purple-red variegation). A total of 181,866 unigenes were generated, including a variety of unigenes involved in anthocyanin biosynthesis and sequestration and the regulation of anthocyanin biosynthesis. The dark purple or purple-red variegation patterns mainly occurred due to the proportions of cyanidin (Cy)- and peonidin (Pn)-based anthocyanins. The variegations of P. rockii exhibited a “Cy > Pn” phenotype, whereas the F1 progeny showed a “Pn > Cy” phenotype. The CHS, DFR, ANS, and GST genes might play key roles in variegation pigmentation in P. rockii according to gene expression and interaction network analysis. Two R2R3-MYB transcription factors (c131300.graph_c0 and c133735.graph_c0) regulated variegation formation by controlling CHS, ANS and GST genes. Our results indicated that the various variegation patterns were caused by transcriptional regulation of anthocyanin biosynthesis genes, and the transcription profiles of the R2R3-MYBs provided clues to elucidate the mechanisms underlying this trait. The petal transcriptome data produced in this study will provide a valuable resource for future association investigations of the genetic regulation of various variegation patterns in tree peonies. Full article

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1897 KiB

Open AccessArticle

Spirulina maxima Extract Prevents Neurotoxicity via Promoting Activation of BDNF/CREB Signaling Pathways in Neuronal Cells and Mice

by Eun-Jeong Koh, Young-Jin Seo, Jia Choi, Hyeon Yong Lee, Do-Hyung Kang, Kui-Jin Kim and Boo-Yong Lee

Molecules 2017, 22(8), 1363; https://doi.org/10.3390/molecules22081363 - 17 Aug 2017

Cited by 35 | Viewed by 8169

Abstract

Spirulina maxima is a microalgae which contains flavonoids and other polyphenols. Although Spirulina maxima 70% ethanol extract (SM70EE) has diverse beneficial effects, its effects on neurotoxicity have not been fully understood. In this study, we investigated the neuroprotective effects of SM70EE against trimethyltin [...] Read more.

Spirulina maxima is a microalgae which contains flavonoids and other polyphenols. Although Spirulina maxima 70% ethanol extract (SM70EE) has diverse beneficial effects, its effects on neurotoxicity have not been fully understood. In this study, we investigated the neuroprotective effects of SM70EE against trimethyltin (TMT)-induced neurotoxicity in HT-22 cells. SM70EE inhibited the cleavage of poly-ADP ribose polymerase (PARP). Besides, ROS production was decreased by down-regulating oxidative stress-associated enzymes. SM70EE increased the factors of brain-derived neurotrophic factor (BDNF)/cyclic AMPresponsive elementbinding protein (CREB) signalling pathways. Additionally, acetylcholinesterase (AChE) was suppressed by SM70EE. Furthermore, we investigated whether SM70EE prevents cognitive deficits against scopolamine-induced neurotoxicity in mice by applying behavioral tests. SM70EE increased step-through latency time and decreased the escape latency time. Therefore, our data suggest that SM70EE may prevent TMT neurotoxicity through promoting activation of BDNF/CREB neuroprotective signaling pathways in neuronal cells. In vivo study, SM70EE would prevent cognitive deficits against scopolamine-induced neurotoxicity in mice. Full article

(This article belongs to the Collection Phytochemicals: Biosynthesis, Metabolism and Biological Activities)

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The effects of SM70EE on cell viability and neuronal death in HT-22 cells. (A) Cell viability was measured by a MTT assay. The experiments were performed in hexa-plicate; (B) HT-22 cells were treated with up to 10 μΜ TMT. Cell lysates were subjected to western blot analysis to analyze to the expression of PARP; (C) HT-22 cells were pre-treated 50 and 100 μg/mL SM70EE for 4 h before treatment with 10 μM TMT for 24 h. Cell lysates were subjected to western blot analysis to probe the expression of PARP. The protein expression level was quantified using Image J software and normalized against α-tubulin and control. Results were analyzed by one-way analysis of variance (ANOVA) and Duncan’s multiple range tests. The p-value in the multiple comparison results (e.g., a, b, and c) indicate significant differences among the groups (p < 0.05). Full article ">Figure 2
SM70EE inhibits ROS production in TMT-induced neurotoxicity in HT-22 cells. (A) HT-22 cells were pretreated 50 and 100 μg/mL SM70EE for 4 h before treatment with 10 μM TMT for 24 h. ROS production was measured by a DCF-DA assay. The experiments were performed in hexa-plicate; (B) HT-22 cells were pre-treated 50 and 100 μg/mL SM70EE for 4 h before treatment with 10 μM TMT for 24 h. Cells lysates were subjected to western blot analysis to measure the levels of HO-1, NOX4, and SOD2. The protein expression level was quantified using Image J software and normalized against α-tubulin and control. Results were analyzed by one-way ANOVA and Duncan’s multiple range tests. The p-value in the multiple comparison results (e.g., a, b, c, and d) indicate significant differences among the groups (p < 0.05). Full article ">Figure 3
SM70EE prevents neuronal cell damage through enhancing BDNF/CREB signaling pathways and suppressing AChE activity against TMT-induced neurotoxicity in HT-22 cells. (A) Proteins were harvested at the indicated time points and subjected to western blot analysis to probe the expressions of p-CREB, p-TrkB; (B) Proteins were harvested at the indicated time points and subjected to western blot to analyze to the expression of BDNF. The protein expression level was quantified using Image J software and normalized against α-tubulin and control; (C) HT-22 cells were pre-treated 50 and 100 μg/mL SM70EE for 4 h before treatment with 10 μM TMT for 24 h. AChE activity was measured by Ellman method. The experiments were performed in tri-plicate. Results were analyzed by one-way ANOVA and Duncan’s multiple range tests. The p-value in the multiple comparison results (e.g., a, b, c, and d) indicate significant differences among the groups (p < 0.05). Full article ">Figure 4
SM70EE prevents learning and memory impairment caused by scopolamine-induced neurotoxicity in mice. (A) Mice were subjected to a passive avoidance test; (B) Mice were subjected to a Morris water maze test. Results were analyzed by one-way ANOVA and Duncan’s multiple range tests. The p-value in the multiple comparison results (e.g., a, b, c, and d) indicate significant differences among the groups (p < 0.05). Full article ">

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Open AccessArticle

Extended Physicochemical Characterization of the Synthetic Anticoagulant Pentasaccharide Fondaparinux Sodium by Quantitative NMR and Single Crystal X-ray Analysis

by William de Wildt, Huub Kooijman, Carel Funke, Bülent Üstün, Afranina Leika, Maarten Lunenburg, Frans Kaspersen and Edwin Kellenbach

Molecules 2017, 22(8), 1362; https://doi.org/10.3390/molecules22081362 - 17 Aug 2017

Cited by 8 | Viewed by 6286

Abstract

Fondaparinux sodium is a synthetic pentasaccharide representing the high affinity antithrombin III binding site in heparin. It is the active pharmaceutical ingredient of the anticoagulant drug Arixtra^®. The single crystal X-ray structure of Fondaparinux sodium is reported, unequivocally confirming both structure [...] Read more.

Fondaparinux sodium is a synthetic pentasaccharide representing the high affinity antithrombin III binding site in heparin. It is the active pharmaceutical ingredient of the anticoagulant drug Arixtra^®. The single crystal X-ray structure of Fondaparinux sodium is reported, unequivocally confirming both structure and absolute configuration. The iduronic acid adopts a somewhat distorted chair conformation. Due to the presence of many sulfur atoms in the highly sulfated pentasaccharide, anomalous dispersion could be applied to determine the absolute configuration. A comparison with the conformation of Fondaparinux in solution, as well as complexed with proteins is presented. The content of the solution reference standard was determined by quantitative NMR using an internal standard both in 1999 and in 2016. A comparison of the results allows the conclusion that this method shows remarkable precision over time, instrumentation and analysts. Full article

(This article belongs to the Special Issue Looking Forward to the Future of Heparin: New Sources, Developments and Applications)

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Structural formula of the synthetic anticoagulant pentasaccharide Fondaparinux sodium. The anomeric D1 and F1 protons used for qNMR integration have been indicated in red. Full article ">Figure 2
Example of the 500 MHz 1H-NMR spectrum of a mixture of the Fondaparinux sodium standard and maleic acid in D2O with the relevant integrated signals. Full article ">Figure 3
Comparison of the observed powder diffraction patterns of amorphous (green) and crystallized Fondaparinux (blue) with the pattern calculated from the single-crystal structure (red). The patterns are given an arbitrary displacement along the intensity axis to aid comparison. For the calculated pattern, the unit-cell parameters were refined using the Rietveld method to allow for the difference in data collection temperature. The reflection positions of observed and calculated patterns show a good correlation. The relative intensities show some differences, which are most likely due to the limited number of crystallites in the powder measurement, some preferred orientation effects and small changes in structure (especially in the water molecules) due to the temperature difference. Full article ">Figure 4
Polarized light microscopic picture at 40× magnification of Fondaparinux crystals formed from a water/ethanol solution. Full article ">Figure 5
Perspective drawing of part of the asymmetric unit of the crystal structure of Fondaparinux. Sodium ions and water molecules (coordinated and non-coordinated) are excluded for clarity. Rings and atoms mentioned in the discussion are labelled. The bonds defining the glycosidic conformation are highlighted in pink. Hydrogen bonds are marked with dashed lines. N3-H3N donates a hydrogen bond to a translation related Fondaparinux molecule (x − 1, y, z − 1). The involved symmetry positions are indicated with a *. Hydroxyl O16 donates a hydrogen bond to a water molecule not shown. Full article ">Figure 6
A comparison of the conformations of the iduronic acid G ring structures in uncomplexed Fondaparinux (a) and Fondaparinux co-crystallized with antithrombin (b); from PDB entry 2GD4) [<a href="#B5-molecules-22-01362" class="html-bibr">5</a>]. Full article ">

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Open AccessArticle

Anticancer Activity of Ramalin, a Secondary Metabolite from the Antarctic Lichen Ramalina terebrata, against Colorectal Cancer Cells

by Sung-Suk Suh, Tai Kyoung Kim, Jung Eun Kim, Ju-Mi Hong, Trang Thu Thi Nguyen, Se Jong Han, Ui Joung Youn, Joung Han Yim and Il-Chan Kim

Molecules 2017, 22(8), 1361; https://doi.org/10.3390/molecules22081361 - 17 Aug 2017

Cited by 28 | Viewed by 7586

Abstract

Colorectal cancer is a leading cause of death worldwide and occurs through the highly complex coordination of multiple cellular pathways, resulting in carcinogenesis. Recent studies have increasingly revealed that constituents of lichen extracts exhibit potent pharmaceutical activities, including anticancer activity against various cancer [...] Read more.

Colorectal cancer is a leading cause of death worldwide and occurs through the highly complex coordination of multiple cellular pathways, resulting in carcinogenesis. Recent studies have increasingly revealed that constituents of lichen extracts exhibit potent pharmaceutical activities, including anticancer activity against various cancer cells, making them promising candidates for new anticancer therapeutic drugs. The main objective of this study was to evaluate the anticancer capacities of ramalin, a secondary metabolite from the Antarctic lichen Ramalina terebrata, in the human colorectal cancer cell line HCT116. In this study, ramalin displayed concentration-dependent anticancer activity against HCT116 cells, significantly suppressing proliferation and inducing apoptosis. Furthermore, ramalin induced cell cycle arrest in the gap 2/mitosis (G2/M) phase through the modulation of hallmark genes involved in the G2/M phase transition, such as tumour protein p53 (TP53), cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin-dependent kinase 1 (CDK1) and cyclin B1 (CCNB1). At both the transcriptional and translational level, ramalin caused a gradual increase in the expression of TP53 and its downstream gene CDKN1A, while decreasing the expression of CDK1 and CCNB1 in a concentration-dependent manner. In addition, ramalin significantly inhibited the migration and invasion of colorectal cancer cells in a concentration-dependent manner. Taken together, these data suggest that ramalin may be a therapeutic candidate for the targeted therapy of colorectal cancer. Full article

(This article belongs to the Collection Efficient Pharmaceutical and Chemical Approaches for Anticancer Therapy: Design, Preliminary Evaluations, and Further Developments)

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The chemical structure of ramalin. Full article ">Figure 2
Effect of ramalin on cell proliferation in human colorectal cancer cells (HCT116). Ramalin significantly suppressed proliferation of HCT116 cells in a time-dependent manner at the highest concentrations used (50 and 100 μg/mL) (A) relative to the untreated control cells. Ramalin treatment significantly decreased the number of colonies in a concentration-dependent manner (B) relative to the untreated control cells. Full article ">Figure 3
Effect of ramalin on the cell cycle in HCT116 cells. Ramalin induced cell cycle arrest in the gap 2/mitosis (G2/M) phase in a concentration-dependent manner (A,B), relative to the untreated control cells. Full article ">Figure 4
Expression of genes involved in the G2/M transition in HCT116 cells. At both translational (A) and transcriptional (B) levels, ramalin significantly increased the expression of TP53 and p21 in a concentration-dependent manner, whereas cyclin B1 and CDK1 expression was significantly decreased. Full article ">Figure 5
Ramalin-mediated apoptosis in HCT116 cells. Ramalin induced apoptosis at the highest concentration used (100 μg/mL) significantly increased the proportion of apoptotic cells relative to that in the untreated control cells. Full article ">Figure 6
Effect of ramalin on wound healing in HCT116 cells. Ramalin significantly suppressed the wound-healing capacity of HCT116 cells in a concentration-dependent manner, relative to the untreated control cells. Full article ">Figure 7
Anti-invasive and migratory activity of ramalin in HCT116 cells. Ramalin significantly suppressed invasion (A) and migration (B) of HCT116 colorectal cancer cells in a concentration-dependent manner (C), relative to the untreated control cells. Full article ">

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Open AccessReview

Bioactive Compounds from Mexican Varieties of the Common Bean (Phaseolus vulgaris): Implications for Health

by Celia Chávez-Mendoza and Esteban Sánchez

Molecules 2017, 22(8), 1360; https://doi.org/10.3390/molecules22081360 - 17 Aug 2017

Cited by 113 | Viewed by 15074

Abstract

As Mexico is located within Mesoamerica, it is considered the site where the bean plant originated and where it was domesticated. Beans have been an integral part of the Mexican diet for thousands of years. Within the country, there are a number of [...] Read more.

As Mexico is located within Mesoamerica, it is considered the site where the bean plant originated and where it was domesticated. Beans have been an integral part of the Mexican diet for thousands of years. Within the country, there are a number of genotypes possessing highly diverse physical and chemical properties. This review describes the major bioactive compounds contained on the Mexican varieties of the common bean. A brief analysis is carried out regarding the benefits they have on health. The effect of seed coat color on the nutraceutical compounds content is distinguished, where black bean stands out because it is high content of anthocyanins, polyphenols and flavonoids such as quercetin. This confers black bean with an elevated antioxidant capacity. The most prominent genotypes within this group are the “Negro San Luis”, “Negro 8025” and “Negro Jamapa” varieties. Conversely, the analyzed evidence shows that more studies are needed in order to expand our knowledge on the nutraceutical quality of the Mexican bean genotypes, either grown or wild-type, as well as their impact on health in order to be used in genetic improvement programs or as a strategy to encourage their consumption. The latter is based on the high potential it has for health preservation and disease prevention. Full article

(This article belongs to the Special Issue Plant Polyphenols as Potential Therapeutic Agents for Diabetes and Obesity)

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Molecules, Volume 22, Issue 8 (August 2017) – 149 articles

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