Differential alkaloid profile in <Emphasis Type="Italic">Uncaria tomentosa</Emphasis> micropropagated plantlets and root cultures

The alkaloids of Uncaria tomentosa micropropagated plantlets and root cultures were isolated and identified by NMR and mass spectrometry. Plantlets yielded pteropodine (1), isopteropodine (2), mitraphylline (3), isomitraphylline (4), uncarine F (5), speciophylline (6), rhynchophylline (7) and isorhynchophylline (8). In plantlets growing under continuous light, tetracyclic alkaloids 7 and 8 decreased from 20 ± 1.8 at 2 months to 2.2 ± 0.33 mg/g dry wt at 6 months, while the pentacyclic alkaloids 1–4 increased from 7.7 ± 1.4 to 15 ± 0.05 mg/g dry wt, supporting their biogenetic conversion. Micropropagated plantlets produced four times more alkaloids (27.6 ± 3.1 mg/g dry wt) than greenhouse plants. Plantlet roots yielded 3, 4, 8 and the glucoindole alkaloids 3α-dihydrocadambine (9) and dolichantoside (10), the last one not previously found in Uncaria.     

Allometric biomass,nutrient and carbon stock models for <Emphasis Type="Italic">Kandelia candel</Emphasis> of the Sundarbans,Bangladesh

Key message

Present study recommends DBH as independent variable of the derived allometric models and Biomass = a + b DBH ² has been selected for total above-ground biomass, nutrients and carbon stock.

Abstract

Kandelia candel (L.) Druce is a shrub to small tree of the Sundarbans mangrove forest of Bangladesh. The aim of the study was to derive the allometric models for estimating above-ground biomass, nutrient and carbon stock in K. candel. A total of eight linear models with 64 regression equations were tested to derive the allometric models for biomass of each part of plant; and nutrients and carbon stock in total above-ground biomass. The best fitted allometric models were selected by considering the values of R ², CV, R _mse, MS_error, S _a, S _b, F value, AICc and Furnival Index. The selected allometric models were Biomass = 0.014 DBH² + 0.03; √Biomass = 0.29 DBH ? 0.21; √Biomass = 0.66 √DBH ? 0.57; √Biomass = 1.19 √DBH ? 1.02; Biomass = 0.21 DBH² + 0.12 for leaves, branches, bark, stem without bark and total above-ground biomass, respectively. The selected allometric models for Nitrogen, Phosphorous, Potassium and Carbon stock in total above-ground biomass were N = 0.39 DBH² + 0.49, P = 0.77 DBH² + 0.14, K = 0.87 DBH² + 0.07 and C = 0.09 DBH² + 0.05, respectively. The derived allometric models have included DBH as a single independent variable, which may give quick and accurate estimation of the above-ground biomass, nutrient and carbon stock in this species. This information may also contribute to a broader study of nutrient cycling, nutrient budgeting and carbon sequestration of the studied forest.

     

Bioconversion of α-pinene by a novel cold-adapted fungus <Emphasis Type="Italic">Chrysosporium pannorum</Emphasis>

The psychrotrophic fungus Chrysosporium pannorum A-1 is reported for the first time as a novel biocatalyst for O₂-promoted oxidation of α-pinene. GC–MS analysis indicated that the main products of the reaction were compounds of a high commercial value, verbenol (1) and verbenone (2). Exponentially growing cells (days 2–3) were about twice as active as cells in the late stationary phase in terms of the total concentration of products. The highest yields of 1 and 2 were obtained using three-day and two-day-old mycelia and a medium containing 1.5 and 1 % (v/v) of the substrate, respectively. The optimal time for the bioconversion of α-pinene varied from 1 to 3 days, and depended on the kind of product desired. Most of 1 was produced at a relatively high concentration of 360 mg/L after the first six hours of α-pinene bioconversion [with an average yield of 69 mg/(g _{dry cell} L aqueous phase)]. The oxidative activity of C. pannorum was identified across a wide temperature range of 5–25 °C, 10 °C being the optimum for the production of 1 and 20 °C for the production of 2. Sequential addition of the substrate during 3 days of the biotransformation resulted in a significant increase in 1 and 2 up to 722 and 176 mg/L, respectively, and a 2-fold enhancement of product yield as compared to bioconversion with a single supply of α-pinene. The concentration of total conversion products in the culture medium reached 1.33 g/L [which corresponded product yield of 225 mg/(g _{dry cell} L)]. This represents probably the most promising result reported to date for oxidative biotransformation of α-pinene by a wild-type microorganism.     

Effect of factor XIII levels and polymorphisms on the risk of myocardial infarction in young patients

The aim of this work was the synthesis, characterization, and cytotoxicity evaluation of three new Ru(II) complexes with a general formula [Ru(Spy)(bipy)(P-P)]PF₆ [Spy = pyridine-6-thiolate; bipy = 2,2′-bipyridine; P-P = 1,2-bis(diphenylphosphine)ethane (1); 1,3-bis(diphenylphosphine) propane (2); and 1,1′-bis(diphenylphosphino)ferrocene] (4). Complex (3) with the 1,4-bis(diphenylphosphine)butane ligand, already known from the literature, was also synthesized, to be better studied here. The cytotoxicities of the complexes toward two kinds of cancerous cells (K562 and S-180 cells) were evaluated and compared to normal cells (L-929 and PBMC) by MTT assay. The complex [Ru(Spy)(bipy)(dppb)]PF₆ (3) was selected to study both the cellular and molecular mechanisms underlying its promising anticancer action in S-180 cells. The results obtained from this study indicated that complex (3) induces cell cycle arrest in the G0/G1 phase in S-180 cells associated with a decrease in the number of cells in S phase. After 24 and 48 h of exposure to complex (3), the cell viability decreased when compared to the negative control. Complex (3) does not appear to be involved in the DNA damage, but induced changes in the mitochondrial membrane potential in S-180 cells. Furthermore, there was also an increase in the gene expression of Bax, Caspase 9, and Tp53. According to our results, complex (3) induces cell apoptosis through p53/Bax-dependent intrinsic pathway and suppresses the expression of active antiapoptotic Bcl-2 protein.     

Tyrosinase and catechol oxidase activity of copper(I) complexes supported by imidazole-based ligands: structure–reactivity correlations

Four new imidazole-based ligands, 4-((1H-imidazol-4-yl)methyl)-2-phenyl-4,5-dihydrooxyzole (L _OL 1), 4-((1H-imidazol-4-yl)methyl)-2-(tert-butyl)-4,5-dihydrooxyzole (L _OL 2), 4-((1H-imidazol-4-yl)methyl)-2-methyl-4,5-dihydrooxyzole (L _OL 3), and N-(2,2-dimethylpropylidene)-2-(1-trityl-1H-imidazol-4-yl-)ethyl amine (L _imz 1), have been synthesized. The corresponding copper(I) complexes [Cu(I)(L _OL 1)(CH₃CN)]PF₆ (CuL _OL 1), [Cu(I)(L _OL 2)(CH₃CN)]PF₆ (CuL _OL 2), [Cu(I)(L _OL 3)(CH₃CN)]PF₆ (CuL _OL 3), [Cu(I)(L _imz 1)(CH₃CN)₂]PF₆ (CuL _imz 1) as well as the Cu(I) complex derived from the known ligand bis(1-methylimidazol-2-yl)methane (BIMZ), [Cu(I)(BIMZ)(CH₃CN)]PF₆ (CuBIMZ), are screened as catalysts for the oxidation of 3,5-di-tert-butylcatechol (3,5-DTBC-H₂) to 3,5-di-tert-butylquinone (3,5-DTBQ). The primary reaction product of these oxidations is 3,5-di-tert-butylsemiquinone (3,5-DTBSQ) which slowly converts to 3,5-DTBQ. Saturation kinetic studies reveal a trend of catalytic activity in the order CuL _OL 3 ≈ CuL _OL 1 > CuBIMZ > CuL _OL 2 > CuL _imz 1. Additionally, the catalytic activity of the copper(I) complexes towards the oxygenation of monophenols is investigated. As substrates 2,4-di-tert-butylphenol (2,4-DTBP-H), 3-tert-butylphenol (3-TBP-H), 4-methoxyphenol (4-MeOP-H), N-acetyl-l-tyrosine ethyl ester monohydrate (NATEE) and 8-hydroxyquinoline are employed. The oxygenation products are identified and characterized with the help of UV/Vis and NMR spectroscopy, mass spectrometry, and fluorescence measurements. Whereas the copper complexes with ligands containing combinations of imidazole and imine functions or two imidazole units (CuL _imz 1 and CuBIMZ) are found to exhibit catalytic tyrosinase activity, the systems with ligands containing oxazoline just mediate a stoichiometric conversion. Correlations between the structures of the complexes and their reactivities are discussed.     

Aromaticity of graphene nanoflakes in a new way: fragment analysis by combination of the nucleus-independent chemical shifts and the anisotropy of current induced density

A graphene nanoflake (GNF) is a polycyclic aromatic hydrocarbon (PAH) with a huge two-dimensional π-conjugated carbon material in which a central benzene ring is surrounded by identical benzene-type rings through infinite alternant method. In this paper, we explore the structure-aromaticity relationship of the GNFs and the GNFs with hollow sites (GNFHs) by combining the nucleus-independent chemical shifts (NICS) with the anisotropy of the current induced density (ACID). Firstly, the benzene is a typical aromatic molecule (NICS = ?9.671 ppm), GNFs 1-6 is darned with benzene and the corresponding GNFHs 1′-6′. Secondly, the NICS values of GNFs 1-6 alternately vary: ?1.214 (1) > ?13.847 (2) < ?2.662 (3) > ?14.530 (4) < ?3.932 (5) > ?13.978 (6) ppm, the GNFs (2, 4, 6) with even fragments of annulene have larger aromaticity than that of GNFs (1, 3, 5) with odd fragments of annulene. Significantly, the NICS values of GNFs 1-6 can also be fragment analyzed by the NICS values and ACID of benzene and corresponding GNFHs 1′-6′. The NICS values for GNFs (2, 4, 6) can be roughly estimated by the NICS value of benzene minus the NICS value of the GNFHs (2′, 4′, 6′), respectively. The NICS values for GNFs (1, 3, 5) can be roughly estimated by the NICS value of the GNFHs (1′, 3′, 5′) minus the NICS value of benzene, respectively. We hope that the present work can provide a simple and reliable method for the rational design of the GNF with aromaticity, which may be used to understand the origin of the graphene nanoflake aromatic properties.     

Highly enantioselective production of a chiral intermediate of sitagliptin by a novel isolate of <Emphasis Type="Italic">Pseudomonas pseudoalcaligenes</Emphasis>

Objective

To produce (S)-3-hydroxy-1-(3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-4-(2,4,5-trifluorophenyl)butan-1-one (S)-1 from 4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro [1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)-1-(2,4,5-trifluorophenyl)butan-2-one (2) by microbial bioreduction.

Results

A new isolate of Pseudomonas pseudoalcaligenes reduced enantioselectively prochiral ketone 2 to chiral alcohol (S)-1. Whole cells of the bacterium were tolerant towards 20 % (v/v) DMSO and 10 g 2/l. Under the optimal conditions, the preparative-scale bioreduction yielded (S)-1 at 90 % yield and >99 % ee. Cells could be re-used with the yield and ee of product being 45 % and >99 %, respectively, after five cycles.

Conclusion

Bioreduction using whole cells of P. pseudoalcaligenes is an attractive approach to produce (S)-1, as a chiral intermediate of the anti-diabetic drug, sitagliptin.

     

An optical material for the detection of trace S<Subscript>2</Subscript>O<Subscript>3</Subscript><Superscript>2−</Superscript> in milk based on a copper complex

A novel S₂O₃ ^2? luminescent sensor (Cu²⁺-p-CPIP) was developed and the presence of S₂O₃ ^2? caused an obvious fluorescence enhancement at 420 nm upon excitation at 330 nm, which could be distinguished with the naked eye under a UV lamp. Remarkably, the compound exhibited excellent selective and sensitive response to S₂O₃ ^2? over other common anions with a micromolar limit of detection (0.442 μM) in DMSO/H₂O (v/v, 1:1) buffer. The absorbance intensity and the color of Cu²⁺-p -CPIP solution changed gradually with the increase of S₂O₃ ^2? concentration. The proposed method was applied to the determination of S₂O₃ ^2? in milk samples and the recoveries were 97.5–105%. The preparation of Cu²⁺-p -CPIP exhibited the quick, simple and facile advantages. The results showed that Cu²⁺-p -CPIP can be a good candidate for simple, rapid and sensitive colorimetric detection of S₂O₃ ^2? in aqueous solution.     

The induction of apoptosis in SGC-7901 cells through the ROS-mediated mitochondrial dysfunction pathway by a Ir(III) complex

A new ligand BTCP and its iridium(III) complex [Ir(ppy)₂(BTCP)]PF₆ (Ir-1) were synthesized and characterized by elemental analysis, ESI–MS, IR, ¹H NMR and ¹³C NMR. The cytotoxic activity in vitro of the ligand and its complex against SGC-7901, HeLa, HOS, PC-12, BEL-7402, MG-63, SiHa, A549, HepG2 and normal cell LO2 were evaluated by MTT method [MTT = (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)]. The apoptosis was assayed with AO/EB and Hoechst 33258 staining methods. The reactive oxygen species (ROS), mitochondrial membrane potential, autophagy and cell invasion were studied under fluorescent microscope. The expression of caspases and Bcl-2 family proteins were investigated by western blot. The IC₅₀ values of complex toward SGC-7901, BEL-7402 and MG-63 cells are 3.9 ± 0.5, 5.4 ± 1.2 and 4.2 ± 0.6 µM. The complex can increase the levels of ROS, and induce a decrease in the mitochondrial membrane potential. Ir-1 inhibits the cell growth at G0/G1 phase in SGC-7901 cells, and the complex can induce both autophagy and apoptosis and inhibit the cell invasion. And the complex induces apoptosis through a ROS-mediated mitochondrial dysfunction pathway.     

Comparative reproductive study of two <Emphasis Type="Italic">Streptocephalus sirindhornae</Emphasis> (Branchiopoda: Anostraca) populations and F1 hybrids: a laboratory study

We investigated the effects of inbreeding and crossbreeding on the reproductive biology of two populations of fairy shrimp Streptocephalus sirindhornae from Maha Sarakham (M) and Suphan Buri (S) provinces of Thailand. Four groups of the mature fairy shrimp were experimentally mated; S × S, M × M, M × S and S × M. The data on life span, age at first spawning, number of eggs per brood and total number of eggs per female of the inbred and crossbred parent generations (P) were not significantly different. The spawning frequency and capacity of the crossbred P were significantly more productive than those of the inbred. In both populations, the majority of P had the greatest egg numbers in the 8th to 14th broods. The hatching percentages of eggs and the survival percentages of nauplii from the 10th brood of the F ₁ populations were significantly higher than those of the 20th and 1st broods. The inbreeding in F ₁ had no effect on hatching and survival percentages, sex ratios of males to females, average weight, body length and specific growth percentages. Our findings suggest that the inbred populations in P and F ₁ were able to yield eggs similar to the crossbred populations, with similar growth, hatching and survival percentages.     

Regional chronologies of <Emphasis Type="Italic">Cedrela</Emphasis><Emphasis Type="Italic">fissilis</Emphasis> and <Emphasis Type="Italic">Cedrela angustifolia</Emphasis> in three forest types and their relation to climate

Key message

Although tree-ring chronologies of Cedrela fissilis and Cedrela angustifolia showed a common climatic signal, local conditions influence growth, suggesting that forest guidelines should be appropriate to the species and context.

Abstract

Cedrela species are highly valued because of the quality of their timber. Understanding the behaviour of each different Cedrela species and their ecology is of importance to ensuring that forest harvesting and management do not endanger the survival of natural populations. These species grow in a wide range of environmental gradients and different types of forests in Bolivia. This study used dendrochronological methods to analyse growth–precipitation relationships of two Cedrela species coming from three locations with different environmental conditions: dry Chiquitano (Concepción), Chiquitano transitional Amazonian (Guarayos), and Bolivian-Tucuman montane forests (Postrervalle). The rainy season in all locations runs from October to April and the dry season runs from May to September. Twelve Cedrela fissilis specimens were sampled from dry Chiquitano, 11 Cedrela fissilis specimens from Chiquitano transitional Amazonian, and 30 Cedrela angustifolia specimens from Bolivian-Tucuman montane forests. The samples were crossdated and exhibited a common signal between trees from three sites, despite tree rings from the Chiquitano transitional Amazonian forest being narrower and displaying blurred bands of parenchyma in the boundaries. Significant inter-series correlation was found for the C. fissilis species series from dry Chiquitano with r = 0.261 (p < 0.01) and Chiquitano transitional Amazonian forests with r = 0.284 (p < 0.01), and for Cedrela angustifolia from Bolivian-Tucuman montane forests with r = 0.374 (p < 0.01). Mean annual growth was 2.07, 1.92, and 2.82 mm year^?1 at the three sites, respectively. Cedrela species from dry Chiquitano and Bolivian-Tucuman montane forests were sensitive to precipitation from October to April of the current growth year (wettest season) and to low temperatures from May to July of the current growth year (driest season). Samples from Chiquitano transitional Amazonian were more sensitive to precipitation during late rainy season (March, April, and May of the current growth year) and high temperatures during the rainy months (November–December). Growth differences between sites and species in response to climate variations and local conditions should be taken into account and handled with different forest management guidelines.

     

How are anatomical and hydraulic features of the mangroves <Emphasis Type="Italic">Avicennia marina</Emphasis> and <Emphasis Type="Italic">Rhizophora mucronata</Emphasis> influenced by siltation?

Key message

This article provides significant data in the debate on whether siltation might have a negative impact on the hydraulic functioning of two widespread mangrove tree species Avicennia marina and Rhizophora mucronata.

Abstract

Elevated sediment addition, or siltation, within mangrove ecosystems is considered as being negative for trees and saplings, resulting in stress and higher mortality rates. However, little is known about how siltation influences the hydraulic functioning of mangrove trees. Comparing two mangrove tree species (Avicennia marina Vierh. Forsk. and Rhizophora mucronata Lam.) from low and high-siltation plots led to the detection of anatomical and morphological differences and tendencies. Adaptations to high siltation were found to be either mutual among both species, e.g., significant smaller single leaf area (p _A.marina  = 0.058, F1.38 = 3.8; p _R.mucronata  = 0.005, F1.38 = 8.7; n = 20 × 20) and a tendency towards smaller stomatal areas (p _A.marina  = 0.131, F1.8 = 2.8; p _R.mucronata  = 0.185, F1.8 = 2.1, n = 5 × 60), or species-specific trends for A. marina, such as higher phloem band/growth layer ratios (p = 0.101, F1.8 = 3.4, n = 5 × 3) and stomatal density (p = 0.052, F1.8 = 5.2, n = 5 × 4). All adaptations seemingly contributed to a comparable hydraulic conductivity independent of the degree of siltation. These findings indicate that silted trees level off fluctuations in their hydraulic performance as a survival mechanism to cope with this less favourable environment. Most of the trees’ structural adaptations to cope with siltation are similar to known drought stress-imposed adaptations.

     

Genetic diversity and population structure of <Emphasis Type="Italic">Melia azedarach</Emphasis> in North-Western Plains of India

Key message

Genetic structure among M. azedarach populations was detected and two subpopulations were present among them. A significant ‘isolation by distance’ was found in M. azedarach population in North-Western Plains of India.

Abstract

Melia azedarach is an important forest tree with pharmaceutical, insecticidal, pesticidal, and commercial significance. It is a good reforestation tree because of its fast growth and drought hardy nature. Genetic variation in a species allows itself to adapt, evolve and respond to environmental stress. It provides the basis for survival of a species and critically influences its evolutionary potential. Assessment of genetic diversity is necessary for improvement and conservation of a species. For this, microsatellite markers are of particular interest given the attributes like co-dominance, reproducibility, hyper variability and abundance throughout the genome. In the present study, we analyzed the genetic diversity and population structure of M. azedarach, an ecologically imperative species growing in the North-Western Plains of India. We developed 43 microsatellite markers, of which 20 were subsequently employed for analysis of diversity and population structure among 33 populations encompassing 318 genotypes representing North-Western Plains of India. A moderate level of diversity (Na = 5.1, Ho = 0.506, He = 0.712, I = 1.386) was assessed. The highest value of ΔK estimated using STRUCTURE indicated 2 subpopulations (K = 2). AMOVA exhibited 73 % variation within populations and 12 % variation was found among regions. Significant positive correlation between geographical and genetic distance was found (Rxy = 0.365, P = 0.010). The present study lays a foundation on a better understanding of genetic dynamics of the species and reveals its diversity and population structure in North-Western Plains of India.

     

In silico study of carvone derivatives as potential neuraminidase inhibitors

Recent outbreaks of highly pathogenic influenza strains have highlighted the need to develop new anti-influenza drugs. Here, we report an in silico study of carvone derivatives to analyze their binding modes with neuraminidase (NA) active sites. Two proposed carvone analogues, CV(A) and CV(B), with 36 designed ligands were predicted to inhibit NA (PDB ID: 3TI6) using molecular docking. The design is based on structural resemblance with the commercial inhibitor, oseltamivir (OTV), ligand polarity, and amino acid residues in the NA active sites. Docking simulations revealed that ligand A18 has the lowest energy binding (?G_bind) value of ?8.30 kcal mol^-1, comparable to OTV with ?G_bind of ?8.72 kcal mol^-1. A18 formed seven hydrogen bonds (H-bonds) at residues Arg292, Arg371, Asp151, Trp178, Glu227, and Tyr406, while eight H-bonds were formed by OTV with amino acids Arg118, Arg292, Arg371, Glu119, Asp151, and Arg152. Molecular dynamics (MD) simulation was conducted to compare the stability between ligand A18 and OTV with NA. Our simulation study showed that the A18-NA complex is as stable as the OTV-NA complex during the MD simulation of 50 ns through the analysis of RMSD, RMSF, total energy, hydrogen bonding, and MM/PBSA free energy calculations.     

Activity of phosphino palladium(II) and platinum(II) complexes against HIV-1 and <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis>

Treatment of human immunodeficiency virus (HIV) is currently complicated by increased prevalence of co-infection with Mycobacterium tuberculosis. The development of drug candidates that offer the simultaneous management of HIV and tuberculosis (TB) would be of great benefit in the holistic treatment of HIV/AIDS, especially in sub-Saharan Africa which has the highest global prevalence of HIV-TB coinfection. Bis(diphenylphosphino)-2-pyridylpalladium(II) chloride (1), bis(diphenylphosphino)-2-pyridylplatinum(II) chloride (2), bis(diphenylphosphino)-2-ethylpyridylpalladium(II) chloride (3) and bis(diphenylphosphino)-2-ethylpyridylplatinum(II) (4) were investigated for the inhibition of HIV-1 through interactions with the viral protease. The complexes were subsequently assessed for biological potency against Mycobacterium tuberculosis H37Rv by determining the minimal inhibitory concentration (MIC) using broth microdilution. Complex (3) showed the most significant and competitive inhibition of HIV-1 protease (p = 0.014 at 100 µM). Further studies on its in vitro effects on whole virus showed reduced viral infectivity by over 80 % at 63 µM (p < 0.05). In addition, the complex inhibited the growth of Mycobacterium tuberculosis at an MIC of 5 µM and was non-toxic to host cells at all active concentrations (assessed by tetrazolium dye and real time cell electronic sensing). In vitro evidence is provided here for the possibility of utilizing a single metal-based compound for the treatment of HIV/AIDS and TB.     

Design and Comparative Evaluation of the Anticonvulsant Profile,Carbonic-Anhydrate Inhibition and Teratogenicity of Novel Carbamate Derivatives of Branched Aliphatic Carboxylic Acids with 4-Aminobenzensulfonamide

Epilepsy is one of the most common neurological diseases, with between 34 and 76 per 100,000 people developing epilepsy annually. Epilepsy therapy for the past 100⁺ years is based on the use of antiepileptic drugs (AEDs). Despite the availability of more than twenty old and new AEDs, approximately 30% of patients with epilepsy are not seizure-free with the existing medications. In addition, the clinical use of the existing AEDs is restricted by their side-effects, including the teratogenicity associated with valproic acid that restricts its use in women of child-bearing age. Thus, there is an unmet clinical need to develop new, effective AEDs. In the present study, a novel class of carbamates incorporating phenethyl or branched aliphatic chains with 6–9 carbons in their side-chain, and 4-benzenesulfonamide-carbamate moieties were synthesized and evaluated for their anticonvulsant activity, teratogenicity and carbonic anhydrase (CA) inhibition. Three of the ten newly synthesized carbamates showed anticonvulsant activity in the maximal-electroshock (MES) and 6 Hz tests in rodents. In mice, 3-methyl-2-propylpentyl(4-sulfamoylphenyl)carbamate(1), 3-methyl-pentan-2-yl-(4-sulfamoylphenyl)carbamate (9) and 3-methylpentyl, (4-sulfamoylphenyl)carbamate (10) had ED₅₀ values of 136, 31 and 14 mg/kg (MES) and 74, 53, and 80 mg/kg (6 Hz), respectively. Compound (10) had rat-MES-ED₅₀?=?13 mg/kg and ED₅₀ of 59 mg/kg at the mouse-corneal-kindling test. These potent carbamates (1,9,10) induced neural tube defects only at doses markedly exceeding their anticonvuslnat-ED₅₀ values. None of these compounds were potent inhibitors of CA IV, but inhibited CA isoforms I, II and VII. The anticonvulsant properties of these compounds and particularly compound 10 make them potential candidates for further evaluation and development as new AEDs.     

Growth and nodulation in <Emphasis Type="Italic">Alnus sieboldiana</Emphasis> in response to <Emphasis Type="Italic">Frankia</Emphasis> inoculation and nitrogen treatments

Key message

We investigated a Frankia – Alnus sieboldiana symbiosis, including the minimum inoculum dose for constant nodulation, the period of time to nodulation after inoculation, and the effects of N on nodulation.

Abstract

Frankia is a nitrogen-fixing actinomycete that forms root nodules in some dicotyledonous plants, which are called actinorhizal. We studied nodule formation in Alnus sieboldiana, an actinorhizal plant, after inoculation with a Frankia isolate to establish techniques for Frankia inoculation and the cultivation of inoculated plants. Root nodules formed on seedlings of A. sieboldiana by 2 weeks after inoculation, and N₂ fixation measured by acetylene reduction activity started 3 weeks after inoculation. Nodulation was observed after inoculation with a Frankia isolate at 0.001 μL packed cell volume (pcv). The number of nodules formed on the seedlings inoculated with Frankia at more than 0.05 μL pcv was not significantly different. Nodule development and N₂ fixation were reduced when inoculated seedlings were treated weekly with 15 mM NH₄NO₃-N. In contrast, treatment with 3.75 or 0.9375 mM NH₄NO₃-N did not inhibit nodule development or N₂ fixation of inoculated seedlings by 15 weeks of N treatment.

     

HUCMNCs protect vascular endothelium and prevent ISR after endovascular interventional therapy for vascular diseases in T2DM rabbits

Secretory phospholipase A₂ (sPLA₂) is a key enzyme participating in the inflammatory cascade followed by the action of cyclooxygenase-2 and lipoxygenases. Therefore, inhibitors of sPLA₂ could be used as potent anti-inflammatory agents to treat the early phase of inflammation. In this study, we have prepared the fenoprofen and ibuprofen analogs containing 1,3,4-oxadiazole nucleus and tested against Vipera russelli venom's basic sPLA₂ (VRV-PL-VIIIa). Among the tested ligands 5(a–t),2-(2-chlorophenyl)-5-(1-(4-phenoxyphenyl) ethyl)-1,3,4-oxadiazole (5m) inhibited the catalytic activity of VRV-PL-VIIIa with an IC₅₀ value of 11.52 µM. Biophysical studies revealed that the 5m quenches the intrinsic fluorescence of VRV-PL-VIIIa, in a concentration dependent manner. Also, the compound 5m affected VRV-PL-VIIIa conformation, which was observed by circular dichroism spectra that recorded the prominent shift in the α-helix peak and the random coil formation of VRV-PL-VIIIa. Further, molecular docking analysis revealed that the compound 5m possess strong hydrophobic interactions at catalytic triad region of the VRV-PL-VIIIa. Evident to in vitro and in silico studies, 5m strongly inhibited the hemolysis of red blood cells. Our in vivo pharmacological studies revealed that the compound 5m inhibited the edematogenic activity of VRV-PL-VIIIa in mouse foot pad. Additionally, the 5m inhibited VRV-PL-VIIIa-induced myotoxicity and lung hemorrhage in mice. Overall, our ADMET results depicted that 5m possess better druggable property. Thus, this study explored the new fenoprofen and ibuprofen analog 5m as the lead-structure that serves as an anti-inflammatory agent.