5-Thioxoimidazolidine-2-one derivatives: Synthesis,anti-inflammatory activity,analgesic activity,COX inhibition assay and molecular modelling study

A series of 5-imino-4-thioxo-2-imidazolidinone derivatives with different substituents at N¹ and N³ was synthesized with high yield and excellent purity by the reaction of different N-arylcyanothioformamide derivatives with isocyanate derivatives. Treatment 5-imino-4-thioxo-2-imidazolidinone derivatives with acidic medium afforded 4-thioxoimidazolidin-2,5-dione derivatives. The structures of the obtained products were established based on spectroscopic IR, ¹H NMR, ¹³C NMR, ¹H, ¹H-COSY, HSQC and elemental analyses. The anti-inflammatory activity of the synthesized compounds through the carrageenan-paw edema model as well as in vitro COX-1 and COX-2 inhibition assay were evaluated where most of the synthesized compounds showed significant anti-inflammatory activity. Mostly, all of our synthesized compounds have greater activity more than celecoxib toward both cyclooxygenase enzymes. All of the tested compounds (except one compound) exhibited IC₅₀ valves for COX-2 ranged from 0.001 × 10⁻³ to 0.827 × 10⁻³ µM while the reference drug has IC₅₀ 40.0 × 10⁻³ µM. Furthermore, the analgesic activity of such compounds was also determined. Molecular modeling study was also conducted to rationalize the potential as anti-inflammatory agents of our synthesized compounds by predicting their binding modes, binding affinities and optimal orientation at the active site of the COX enzymes.     

Binding of lipophorin to the fat body of the migratory locust

The interaction of locust high density lipophorin (HDLp) with pieces of fat body tissue was studied at 33°C using a radiolabelled ligand binding assay. Under the assay conditions, binding of tritium-labelled HDLp ([³H]HDLp) was demonstrated to correlate linearly with tissue concentration up to ∼ 7 mg of fat body protein per ml of incubation medium. The [³H]HDLp binding that was displaceable by a 20-fold excess of unlabelled HDLp (which is an approximation of the specific binding) reached equilibrium after ∼ 2 h, whereas low levels of non-displaceable binding increased linearly during this time interval. Analysis of the concentration dependent total binding of [³H]HDLp revealed the presence of a specific binding site with an equilibrium dissociation constant of K_d = 3.1 (±0.5) × 10⁻⁷ M and a maximal binding capacity of 9.8 (±0.5) ng μg⁻¹ tissue protein. Competition experiments demonstrated that the affinity of unlabelled HDLp for the binding site is similar to the affinity of [³H]HDLp. Unlabelled low density lipophorin (LDLp), however, was shown to have an approx. 20-fold lower affinity for the binding site.     

Stepwise Frontal Analysis Coupled with Affinity Chromatography: A Fast and Reliable Method for Potential Ligand Isolation and Evaluation from Mahuang-Fuzi-Xixin Decoction

Mahuang-Fuzi-Xixin Decoction (MFXD) is widely used in the treatment of asthma, however, the functional components in the decoction targeting beta2-adrenoceptor (β₂-AR) remain unclear. Herein, we immobilized the haloalkane dehalogenase (Halo)-tagged β₂-AR on the 6-chlorocaproic acid-modified microspheres. Using the affinity stationary phase, the interactions of four ligands with the receptor were analyzed by stepwise frontal analysis. The association constants were (4.75±0.28)×10⁴ M⁻¹ for salbutamol, (2.93±0.15)×10⁴ M⁻¹ for terbutaline, (1.23±0.03)×10⁴ M⁻¹ for methoxyphenamine, (5.67±0.38)×10⁴ M⁻¹ for clorprenaline at high-affinity binding site, and (2.73±0.05)×10³ M⁻¹ at low-affinity binding site. These association constants showed the same rank order as the radioligand binding assay, demonstrating that immobilized β₂-AR had capacity to screen bioactive compounds binding to the receptor while stepwise frontal analysis could predict their binding affinities. Application of the immobilized receptor in analysis of MFXD by chromatographic method revealed that ephedrine, aconifine, karakoline, and chasmanine were the bioactive compounds targeting β₂-AR. Among them, ephedrine and chasmanine exhibited association constants of (2.94±0.02)×10⁴ M^-1and (4.60±0.15)×10⁴ M⁻¹ to the receptor by stepwise frontal analysis. Molecular docking analysis demonstrated that ephedrine, chasmanine, and the other two compounds interact with β₂-AR through the same pocket involving the key amino acids such as Asn312, Asp113, Phe289, Trp286, Tyr316, and Val114. As such, we reasoned that the four compounds dominate the therapeutic effect of MFXD against asthma through β₂-AR mediating pathway. This work shed light on the potential of immobilized β₂-AR for drug discovery and provided a valuable methodology for rapid screening.     

Synthesis, in vitro and in vivo biological evaluation,COX-1/2 inhibition and molecular docking study of indole-N-acylhydrazone derivatives

The objective of this work was to obtain and evaluate anti-inflammatory in vitro, in vivo and in silico potential of novel indole-N-acylhydrazone derivatives. In total, 10 new compounds (3a–j) were synthesized in satisfactory yields, through a condensation reaction in a single synthesis step. In the lymphoproliferation assay, using mice splenocytes, 3a and 3b showed inhibition of lymphocyte proliferation of 62.7% (±3.5) and 50.7% (±2), respectively, while dexamethasone presented an inhibition of 74.6% (±2.4). Moreover, compound 3b induced higher Th2 cytokines production in mice splenocytes cultures. The results for COX inhibition assays showed that compound 3b is a selective COX-2 inhibitor, but with less potency when compared to celecoxib, and compound 3a not presented selectivity towards COX-2. The molecular docking results suggest compounds 3a and 3b interact with the active site of COX-2 in similar conformations, but not with the active site of COX-1, and this may be the main reason to the COX-2 selectivity of compound 3b. In vivo carrageenan-induced paw edema assays were adopted for the confirmation of the anti-inflammatory activity. Compound 3b showed better results in suppressing edema at all tested concentrations and was able to induce an edema inhibition of 100% after 5?h of carrageenan injection at the 30?mg?kg^?1 dosage, corroborating with the COX inhibition and lymphoproliferation results. I addition to our experimental results, in silico analysis suggest that compounds 3a and 3b present a well-balanced profile between pharmacodynamics and pharmacokinetics. Thus, our preliminary results revealed the potentiality of a new COX-2 selective derivative in the modulation of the inflammatory process.     

PRODUCTIVITY OF PODOSTEMUM CERATOPHYLLUM IN THE NEW RIVER,VIRGINIA

Productivity of Podostemum ceratophyllum, the dominant aquatic macrophyte in the New River, was measured at four sites representing soft- and hardwater reaches of the river. Available dissolved inorganic carbon (DIC) was 4–5 times greater in the hardwater reach. The difference in available DIC was reflected in standing crop and productivity of P. ceratophyllum. Maximum standing crops of P. ceratophyllum at the two hardwater sites (Sites 1 and 2) were 244.8 ± 30.7 g ash-free dry wt (AFDW) m^−-2 and 193.8 ± 18.7 g AFDW m^−-2 compared to 128.5 ± 14.9 g AFDW m^−-2 and 101.3 ± 6.9 g AFDW m^−-2 for the softwater sites (Sites 3 and 4). Productivity, based on differences in standing crops, was: Site 1, 1.08 ± 0.12 g C m^−-2 d^−-1; Site 2, 0.86 ± 0.08 g Cm^−-2d^−-1; Site 3,0.58 ± 0.06 g C m^−-2 d^−-1; Site 4,0.45 ± 0.03 g C m^−-2 d^−-1. Corresponding values for productivity as ¹⁴C uptake were: 2.77 ± 0.44 g C m^−-2 d^−-1; 2.10 ± 0.45 g C m^−-2 d^−-1; 0.34 ± 0.04 g C m^−-2 d^−-1; 0.28 ± 0.03 g C m^−-2 d^−-1. Productivity/biomass (P/B) based on ¹⁴C uptake and standing crop revealed that P. ceratophyllum productivity was inhibited at the softwater sites perhaps due to carbon limitation. Because of its abundance and its high productivity, P. ceratophyllum is hypothesized to contribute significantly to the New River organic matter budget.     

Anti-inflammatory Effect from a Hydrogel Containing Nanoemulsified Copaiba oil (<Emphasis Type="Italic">Copaifera multijuga</Emphasis> Hayne)

Copaiba oil is used as a popular medicine in the Amazonian forest region, especially due to its anti-inflammatory properties. In this paper, we describe the formulation of hydrogel containing copaiba oil nanoemulsions (with positive and negative charges), its skin permeation, and its anti-inflammatory activity in two in vivo models: mouse ear edema and rat paw edema. Three hydrogels were tested (Carbopol^®, hydroxyethylcellulose and chitosan), but only Carbopol^® and hydroxyethylcellulose hydrogels presented good stability and did not interfere with the nanoemulsions droplet size and polydispersity index. In skin permeation assay, both formulations, positively charged nanoemulsion (PCN) and negatively charged nanoemulsion (NCN), presented a high retention in epidermis (9.76 ± 2.65 μg/g and 7.91 ± 2.46 μg/cm², respectively) followed by a smaller retention in the dermis (2.43 ± 0.91 and 1.95 ± 0.56 μg/cm², respectively). They also presented permeation to the receptor fluid (0.67 ± 0.22 and 1.80 ± 0.85 μg/cm², respectively). In addition, anti-inflammatory effect was observed to NCN and PCN with edema inhibitions of 69 and 67% in mouse ear edema and 32 and 72% in rat paw edema, respectively. Histological cuts showed the decrease of inflammatory factors, such as dermis and epidermis hyperplasia and inflammatory cells infiltration, confirming the anti-inflammatory effect from both copaiba oil nanoemulsions incorporated in hydrogel.     

Proliferation inhibition of novel diphenylamine derivatives

Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most widely used drugs in the world but some NSAIDs such as diclofenac and tolfenamic acid display levels of cytotoxicity, an effect which has been attributed to the presence of diphenylamine contained in their structures. A novel series of diphenylamine derivatives were synthetised and evaluated for their cytotoxic activities and proliferation inhibition. The most active compounds in the cytotoxicity tests were derivative 6g with an IC₅₀ value of 2.5 ± 1.1 × 10⁻⁶ M and derivative 6f with an IC₅₀ value of 6.0 ± 3.0 × 10⁻⁶ M (L1210 cell line) after 48 h incubation. The results demonstrate that leukemic L1210 cells were much more sensitive to compounds 6f and 6g than the HEK293T cells (IC₅₀ = 35 × 10⁻⁶ M for 6f and IC₅₀ > 50 × 10⁻⁶ M for 6g) and NIH-3T3 (IC₅₀ > 50 × 10⁻⁶ M for both derivatives). The IC₅₀ values show that these substances may selectively kill leukemic cells over non-cancer cells. Cell cycle analysis revealed that a primary trend of the diphenylamine derivatives was to arrest the cells in the G₁-phase of the cell cycle within the first 24 h. UV–visible, fluorescence spectroscopy and circular dichroism were used in order to study the binding mode of the novel compounds with DNA. The binding constants determined by UV–visible spectroscopy were found to be in the range of 2.1–8.7 × 10⁴ M⁻¹. We suggest that the observed trend for binding constant K is likely to be a result of different binding thermodynamics accompanying the formation of the complexes.     

Substituted phenyl[(5-benzyl-1,3,4-oxadiazol-2-yl)sulfanyl]acetates/acetamides as alkaline phosphatase inhibitors: Synthesis,computational studies,enzyme inhibitory kinetics and DNA binding studies

Substituted phenyl[(5-benzyl-1,3,4-oxadiazol-2-yl)sulfanyl]acetates/acetamides 9a-j were synthesized as alkaline phosphatase inhibitors. Phenyl acetic acid 1 through a series of reactions was converted into 5-benzyl-1,3,4-oxadiazole-2-thione 4. The intermediate oxadiazole 4 was then reacted with chloroacetyl derivatives of phenols 6a-f and anilines derivatives 8a-d to afford the title oxadiazole derivatives 9a-j. All of the title compounds 9a-j were evaluated for their inhibitory activity against human alkaline phosphatise (ALP). It was found that compounds 9a-j exhibited good to excellent alkaline phosphatase inhibitory activity especially 9h displayed potent activity with IC₅₀ value 0.420 ± 0.012 µM while IC₅₀ value of standard (KH₂PO₄) was 2.80 µM. The enzyme inhibitory kinetics of most potent inhibitor 9h was determined by Line-weaever Burk plots showing non-competitive mode of binding with enzyme. Molecular docking studies were performed against alkaline phosphatase enzyme (1EW2) to check the binding affinity of the synthesized compounds 9a-j against target protein. The compound 9h exhibited excellent binding affinity having binding energy value (−7.90 kcal/mol) compared to other derivatives. The brine shrimp viability assay results proved that derivative 9h was non-toxic at concentration used for enzyme assay. The lead compound 9h showed LD₅₀ 106.71 µM while the standard potassium dichromate showed LD₅₀ 0.891 µM. The DNA binding interactions of the synthesized compound 9h was also determined experimentally by spectrophotometric and electrochemical methods. The compound 9h was found to bind with grooves of DNA as depicted by both UV–Vis spectroscopy and cyclic voltammetry with binding constant values 7.83 × 10³ and 7.95 × 10³ M⁻¹ respectively revealing significant strength of 9h-DNA complex. As dry lab and wet lab results concise each other it was concluded that synthesized compounds, especially compound 9h may serve as lead compound to design most potent inhibitors of human ALP.     

Synthesis and biological evaluation of some thiazole derivatives as new cholinesterase inhibitors

In the present study, some thiazole derivatives were synthesized via the ring closure reaction of 1-[2-(2-oxobenzo[d]thiazol-3(2H)-yl)acetyl]thiosemicarbazide with various phenacyl bromides. The chemical structures of the compounds were elucidated by ¹H NMR, ¹³C NMR and mass spectral data and elemental analyses. Each derivative was evaluated for its ability to inhibit acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) using a modification of Ellman’s spectrophotometric method. The compounds were also investigated for their cytotoxic properties using MTT assay. The most potent AChE inhibitor was found as compound 4e (IC₅₀?=?25.5?±?2.12 µg/mL) followed by compounds 4i (IC₅₀?=?38.50?±?2.12 µg/mL), 4c (IC₅₀?=?58.42?±?3.14 µg/mL) and 4g (IC₅₀?=?68?±?2.12 µg/mL) when compared with eserine (IC₅₀?=?0.025?±?0.01 µg/mL). Effective compounds on AChE exhibited weak inhibition on BuChE (IC₅₀ > 80 µg/mL). MTT assay indicated that the cytotoxic dose (IC₅₀?=?71.67?±?7.63 µg/mL) of compound 4e was higher than its effective dose.     

Citrus Honeys from Three Different Regions of Turkey: HPLC-DAD Profiling and in Vitro Enzyme Inhibition,Antioxidant, Anti-Inflammatory and Antimicrobial Properties with Chemometric Study

The objectives of the present study are to compare the phenolic profiles and biological activities of 15 citrus honey samples from three different locations in Turkey using a chemometric approach. The HPLC-DAD analysis was used to determine phenolic profiles. Nineteen phenolic compounds were identified. Gallic acid (107.14–717.04 μg/g) was recorded as the predominant compound. AF (Antalya-Finike) had the highest antioxidant activity in ABTS⋅⁺ (IC₅₀: 18.01±0.69 mg/mL), metal chelating (IC₅₀: 6.20±0.19 mg/mL) and CUPRAC (A_0.50: 12.05±0.68 mg/mL) assays, while it revealed the best anti-inflammatory activity against COX-2 (17.28±0.22 %) and COX-1 (43.28±0.91 %). AM (Antalya-Manavgat) was the most active in β-carotene-linoleic acid (IC₅₀: 10.05±0.19 mg/mL), anti-urease (38.90±0.69 %), anti-quorum sensing and antimicrobial activities. AKO1 (Adana-Kozan-1) in DPPH⋅ (IC₅₀: 34.25±0.81 mg/mL) assay, AKU1 (Antalya-Kumluca-1) in tyrosinase inhibition activity (37.73±0.38 %) assay, AKU2 (Antalya-Kumluca-2) in AChE (10.55±0.63 %) and BChE (9.18±0.45 %) inhibition activity assays showed the best activity. Chemometric tools were applied to the phenolic compositions and biological properties. PCA and HCA ensured that 15 citrus honey samples were grouped into 3 clusters. The results showed that myricetin, kaempferol, vanillin, protocatechuic acid, rosmarinic acid, rutin, vanillic acid, gallic acid, catechin and p-hydroxyphenyl acetic acid are phenolic compounds that can be used in the classification of citrus honeys.     

Cultivation of Lemna gibba under desert conditions. I: Twelve months of continuous cultivation in open ponds

Duckweed, Lemna gibba, was grown in 12 m² shallow ponds in the Negev desert, during 12 months of continuous cultivation, beginning April 1984. Average monthly growth rates varied with the season of the year. The lowest daily yield, 2·6±0·4 g dry weight m⁻² day⁻¹, was obtained during January. Highest daily yields, 7·9±2·6 g dry weight m⁻² day⁻¹ and 7·0±1·2 g dry weight m⁻² day⁻¹, were obtained during September and May. A 35% decline of the yield was seen during midsummer (July), 4·8±1·2 g dry weight m⁻² day⁻¹. The average rate for the year was 5·15±1·7 g dry weight m⁻² day⁻¹. The protein content of the plants ranged from 30 to 38% per unit dry weight.Growth performance is discussed in relation to the prevailing climatic conditions.     

Evaluation of the effects of different stocking densities on growth and stress responses of juvenile hybrid grouper ♀ Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatus in recirculating aquaculture systems

This study was aimed at evaluating the physiological and metabolic responses of juvenile hybrid grouper ♀ Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatus to stocking density. Hybrid grouper juveniles (mean ± SE = 25.43 ± 2.36 g live mass) were stocked for 22 weeks in a recirculating aquaculture system (RAS) under four different densities: low stocking density (LD; 1.03 kg m⁻³), medium stocking density (MD; 2.06 kg m⁻³), high stocking density (HD; 3.09 kg m⁻³) and extra-high stocking density (EHD; 4.11 kg m⁻³). Biometric variables were recorded and plasma, liver, intestine and stomach samples were taken for biochemical analysis at the end of the experimental period. Final stocking densities were 6.27, 16.04, 23.77 and 28.32 kg m⁻³, respectively, with significant differences in growth performance. Our results showed that the best growth rates and feed utilisation occurred in the MD group. Higher plasma cortisol and glucose levels and lower triglyceride levels reflected the stress responses in the EHD group. Moreover, the activity of aspartate and alanine transaminases was elevated in the HD and EHD groups due to enhanced gluconeogenesis. The activity of the digestive enzyme pepsin significantly increased in the MD group. We found that 2.06–3.09 kg m⁻³ is the most suitable starting density for culturing juvenile hybrid grouper in recirculating aquaculture systems.     

Antiangiogenic,anti-inflammatory and their antioxidant activities of Turnera subulata Sm. (Turneraceae)

Turnera subulata is a substantial medicinal plant used in folk medicine to treat various ailments. The current study was assess the total phenolic and flavonoid contents to evaluate the antioxidant and anti-inflammatory activities of the sequentially extracted T. subulata plant samples. In vitro anti-angiogenic activity was evaluated by chick chorioallantoic membrane (CAM) model for chloroform, ethyl acetate and ethanol extracts. The results obtained revealed that total phenolic content of the chloroform extract (24.13 ± 0.27 mg/g) and total flavonoid content (TFC) of the chloroform extract (22.28 ± 0.40 mg/g) were found to be suggestively higher than the other extracts. A strong antioxidant property was observed for all the six extracts. A study anti-inflammatory activity was observed in chloroform and ethanol extracts, with IC₅₀ ranging from 79 ± 1.01 μg/mL to 81 ± 1.01 μg/mL for protein denaturation assay and from 74 ± 0.11 μg/mL to 76 ± 1.11 μg/mL for HRBC membrane stabilization assay, respectively. The chloroform and ethanol extracts have exhibited good antiangiogenic property. Eventually, these results justified that the chloroform and ethanol extracts of T. subulata with great antioxidant, anti-inflammatory and antiangiogenesis potentials could be promising candidates for the development of a cost effective, potent anticancer drug with minimal side effects.     

In vitro and in silico characterization of angiogenic inhibitors from <Emphasis Type="Italic">Sophora interrupta</Emphasis>

Sophora interrupta Bedd, (Fabaceae) is used in Indian folk medicine to treat cancer. Angiogenesis is one of the crucial characteristics of cancer metastasis and is regulated by vascular endothelial growth factor (VEGF). In this study, we examined the antiangiogenic properties of the root ethyl acetate extract of Sophora interrupta by various methods. In vitro antioxidant activity (100–600 μg/ml) of S. interrupta ethyl acetate (SEA) extract was evaluated by DPPH and ABTS, anti-inflammatory activity (50, 100 and 150 μg/ml) by estimating nitric oxide (NO) levels, anti-angiogenic activity (200 and 500 μg/ml) was validated by chorio allantoic membrane (CAM) assay and in silico molecular dynamic (MD) simulations analyses (25 ns) were performed to identify the anti-angiogenic compounds extracted from root extract. The antioxidative activity of SEA extract at IC₅₀ (200?±?0.6 μg/mL) is equal to that of ascorbic acid at IC₅₀ (50?±?0.6 μg/mL), and the anti-inflammatory activity of SEA extract at IC₅₀ (150?±?0.2 μg/mL) was inhibited significantly by nitric oxide (NO) production. The SEA extract significantly reduced the sprouting of new blood vessels at ID₅₀ 500?±?0.13 μg/mL in the CAM assay. Gas chromatography–mass spectrometry analysis of the SEA extract detected 34 secondary metabolites, of which 6a,12a-dihydro-6H-(1,3)dioxolo(5,6)benzofuro(3,2-c)chromen-3-ol (maackiain) and funiculosin formed strong hydrogen bond interactions with Lys 920, Thr 916 and Cys 919 (2H), as well as Glu 917 of VEGFR2, and these interactions were similar to those of the anti-angiogenic compound axitinib. Significant findings in all the assays performed indicate that SEA extract has potential anti-angiogenic compounds that may interfere with VEGF-induced cancer malignancy.     

Cytotoxicity and comparative binding mechanism of piperine with human serum albumin and α-1-acid glycoprotein

Human serum albumin (HSA) and α-1-acid glycoprotein (AGP) (acute phase protein) are the plasma proteins in blood system which transports many drugs. To understand the pharmacological importance of piperine molecule, here, we studied the anti-inflammatory activity of piperine on mouse macrophages (RAW 264.7) cell lines, which reveals that piperine caused an increase in inhibition growth of inflammated macrophages. Further, the fluorescence maximum quenching of proteins were observed upon binding of piperine to HSA and AGP through a static quenching mechanism. The binding constants obtained from fluorescence emission were found to be K_piperine?=?5.7 ± .2 × 10⁵ M^?1 and K_piperine = 9.3± .25 × 10⁴ M^?1 which correspond to the free energy of ?7.8 and ?6.71 kcal M^?1at 25 °C for HSA and AGP, respectively. Further, circular dichrosim studies revealed that there is a marginal change in the secondary structural content of HSA due to partial destabilization of HSA–piperine complexes. Consequently, inference drawn from the site-specific markers (phenylbutazone, site I marker) studies to identify the binding site of HSA noticed that piperine binds at site I (IIA), which was further authenticated by molecular docking and molecular dynamic (MD) studies. The binding constants and free energy corresponding to experimental and computational analysis suggest that there are hydrophobic and hydrophilic interactions when piperine binds to HSA. Additionally, the MD studies have showed that HSA–piperine complex reaches equilibration state at around 3 ns, which prove that the HSA–piperine complex is stable in nature.     

Pyrazoles containing thiophene,thienopyrimidine and thienotriazolopyrimidine as COX-2 selective inhibitors: Design,synthesis, in vivo anti-inflammatory activity,docking and in silico chemo-informatic studies

New thiophene and annulated thiophene pyrazole hybrids were synthesized and screened for their in vitro COX-1/COX-2 enzymatic inhibition and in vivo anti-inflammatory activities. All compounds were more COX-2 selective inhibitors than COX-1 with compound 13 exhibiting the highest COX-2 selectivity index. Compounds 3, 6a, 9 and 11 were the most promising in the acute anti-inflammatory assay while compounds 3, 5, 6a, 6c, 9, 10, 11 and 13 exerted promising anti-inflammatory activity in the sub-acute anti-inflammatory assay. Compounds 3, 6a, 6c, 9, 10 and 11 were evaluated for their ED₅₀ values and were more potent than diclofenac sodium while compounds 6a, 6c and 9 were of greater potency than celecoxib with compound 6a being the most potent showing ED₅₀ = 0.033 mmol/kg. These compounds were non-toxic and proved to be gastrointestinal safe compared to indomethacin, diclofenac sodium and celecoxib. Docking studies into COX-2 active site (PDB code 3LN1) revealed that compounds 3, 6a, 6c, 9, 10, 11 and 13 had binding modes and energies comparable to that of celecoxib. Compounds 3, 9, 10 and 11 complied with Lipinski’s RO5 while compounds 6a and 6c showed one violation whereas compound 13 deviated by 2 violations. Compounds 6a, 6c and 13 showed 100% plasma protein binding (PPB) and showed no aqueous solubility while compounds 3, 10 and 11 demonstrated the best drug likeness model scores. Therefore, the thiophene analog 3 and the thienopyrimidine derivatives 10 and 11 are promising anti-inflammatory candidates that exert moderate selective COX-2 inhibition with acceptable physicochemical properties.     

Nematicidal and Molecular Docking Investigation of Essential Oils from Pogostemon cablin Ecotypes against Meloidogyne incognita

Root-knot nematode, Meloidogyne incognita is one of the most destructive nematodes worldwide. Essential oils (EOs) are being extensively utilized as eco-benign bionematicides, although the precise mechanism of action remains unclear. Pogostemon cablin Benth. is well-known as “Patchouli”. It is native to South East Asia and known for ethno-pharmacological properties. In this study, chemical composition and potential nematicidal effect of EOs hydrodistilled from the leaves of P. cablin grown at three different locations in India were comprehensively investigated to correlate their mechanism of action for target specific binding affinities toward nematode proteins. Aromatic volatile Pogostemon essential oils (PEO) from Northern India (PEO-NI), Southern India (PEO-SI) and North Eastern India (PEO-NEI) were analyzed by Gas Chromatography-Mass Spectrometry (GC/MS) to characterize forty volatile compounds. Maximum thirty-three components were identified in PEO-NEI. Sesquiterpenes were predominant with higher content of α-guaiene (2.3–24.4 %), patchoulol (6.1–32.7 %) and α-bulnesene (5.9–27.1 %). Patchoulol was the major component in PEO-SI (32.7±1.2 %) and PEO-NEI (29.2±1.1 %), while α-guaiene in PEO-NI (24.4±1.2 %). In vitro nematicidal assay revealed significant nematicidal action (LC₅₀ 44.6–87.0 μg mL⁻¹) against juveniles of M. incognita within 24 h exposure. Mortality increases with increasing time to 48 h (LC₅₀ 33.6–71.6 μg mL⁻¹) and 72 h (LC₅₀ 27.7–61.2 μg mL⁻¹). Molecular modelling and in silico studies revealed multi-modal inhibitive action of α-bulnesene (−22 to −13 kJ mol⁻¹) and α-guaiene (−22 to −12 kJ mol⁻¹) against three target proteins namely, acetyl cholinesterase (AChE), odorant response gene-1 (ODR1), odorant response gene-3 (ODR3). Most preferable binding mechanism was observed against AChE due to pi-alkyl, pi-sigma, and hydrophobic interactions. Structure nematicidal activity relationship suggested the presence of hydroxy group for nematicidal activity is nonessential, rather highly depends on synergistic composition of sesquiterpene hydrocarbons.     

N2(C2H2)ASE ACTIVITY BY ALNUS INCANA SSP. RUGOSA (BETULACEAE) IN THE NORTHERN HARDWOOD FOREST

Field assays of N₂(C₂H₂)ase activity were performed with intact nodules from a pure alder site (alder) and a mixed alder-aspen site (aspen). Assays were performed between 12 June and 12 August 1980 and in May 1981. N₂(C₂H₂)ase rates are expressed as g N g nodule oven-dry wt⁻¹ hr⁻¹ (g N g⁻¹ hr⁻¹). Diurnal N₂(C₂H₂)ase activity showed an increase in both sites between 0600 and midday, then decreased to a low by 1800. Nighttime activity in the May 1981 assay was approximately 25% of the daytime peak. Mean (±SE) 1200 hr N₂(C₂H₂)ase activity (μg N g⁻¹ hr⁻¹) for all sizes in the alder stand rose from 24.56 ± 6.56 on 12 June to 73.96 ± 28.37 on 26 June and declined to 9.20 ± 2.56 by 12 August. In the aspen stand activity decreased from the 12 June rate of 21.81 ± 4.59 to 3.64 ± 1.87 on 24 July but then increased to 30.00 ± 7.39 by 12 August. Based on diurnal assays, the seasonal mean N influx (μg N g⁻¹ hr⁻¹) is statistically higher (P 0.05) in the alder stand with a value of 26.70 compared to 14.63 in the aspen stand. Small size class shrubs had significantly higher (P < 0.05) N₂(C₂H₂)ase activity (μg N g⁻¹ hr⁻¹) in diurnal assays than medium or large class shrubs. The estimated mean (±SE) N₂(C₂H₂)ase activity (mg N g⁻¹ season⁻¹) for all sizes was 44.4 ± 18.6 in the alder stand compared to 16.2 ± 5.2 in the aspen stand. Nodule excavations showed the g shrub⁻¹ in the alder stand to be 16.48 ± 10.29, 38.57 ± 12.34 and 29.11 ± 7.15 for small, medium and large size shrubs and 12.73 ± 3.23, 28.21 ± 4.36 and 56.45 ± 16.23 for respective sizes in the aspen stand. Seasonal N influx was 4.69 kg ha⁻¹ in the alder stand and 0.84 kg ha⁻¹ in the aspen stand, representing 17.9% of the alder stand. Nitrogen feedback inhibition from uric acid-N influx and allelochemic interference from aspen are discussed as explanations for the differences in N influx in the two stands.     

Typha productivity in a Texas pond: Implications for energy and nutrient dynamics in freshwater wetlands

Above-and below-ground biomass of Typha angustifolia L. was sampled monthly for 18 months from a small Texas pond. Maximum above-ground biomass was 2559±284 g AFDW (ash-free dry weight) m⁻² in 1983 and 2895±217 g AFDW m⁻² in 1984. Peak below-ground biomass for these 2 years was 2506±278 g AFDW m⁻² and 2314±226 g AFDW m^t-2, respectively. Stepwise multiple linear regression analyses revealed that mean above-ground biomass accrual was related to duration of growing season, cumulative precipitation, cumulative degree days and/or cumulative pan evaporation. The same was not true for below-ground biomass increases. Analysis of covariance revealed that the rates of above-ground biomass production were not significantly different (F test, p < 0.05) between the 1983 and 1984 growing seasons. Below-ground biomass turnover times for 1983 and 1984 were 2.47 and 1.21 years, respectively.