A new species, Trichiurus australis (Perciformes: Trichiuridae), from Australia

Trichiurus australis, a new trichiurid fish, is described on the basis of four specimens collected off Burnett River mouth, Pialba and Thursday Island, Queensland, Australia. The new species strongly resembles Trichiurus brevis Wang and You, 1992 (distributed in the South China Sea), T. russelli Dutt and Thankam, 1967 (the northwestern Bay of Bengal and the northern Gulf of Thailand), and T. nickolensis Burhanuddin and Iwatsuki, 2003 (northwestern Australia) in having the highest point of the supraoccipital crest situated directly above the posterior margin of the eye and being relatively small in size (less than ca. 700mm in total length: TL). It differs from those three species in having spinescent gill rakers almost with 2 equally long cusps [vs. (1, rarely 2 but 1 of them clearly shorter than another)], posterior caudal peduncle vertebrae bearing neural spines (vs. neural spines absent), longer caudal peduncle length (mean 16% TL vs. 6% in T. brevis, 8% in T. nickolensis, and 8% in T. russelli), and shorter precaudal length (83% vs. 93%, 91%, and 91%), preanal length (26% vs. 33%, 31%, and 35%), and head length (9% vs. 12%, 11%, and 13%). We have tentatively classified this new species under the Trichiurus russelli complex because the foregoing diagnostic characters are identical to this species complex except for the presence of neural spines in the posterior caudal vertebrae.     

Vitamin E inhibits hepatic oxidative stress, toxicity and hyperproliferation in rats treated with the renal carcinogen ferric nitrilotriacetate

Ferric nitrilotriacetate (Fe-NTA) is a potent renal and hepatic tumor promoter, which acts through a mechanism involving oxidative stress. Fe-NTA when injected intraperitoneally into rats induces hepatic ornithine decarboxylase activity as well as hepatic DNA synthesis. Vitamin E is a well-known, lipid-soluble and chain-breaking antioxidant which protects cell membranes from peroxidative damage. In this study, we investigated the protective effect of vitamin E, a major fat-soluble antioxidant, against Fe-NTA-mediated hepatic oxidative stress, toxicity and hyperproliferation in Wistar rats. Animals were treated with two different doses of vitamin E for 1 week prior to Fe-NTA treatment. Vitamin E at a higher dose of 2.0 mg/animal/day showed significant reduction in Fe-NTA-induced hepatic ornithine decarboxylase activity, DNA synthesis, microsomal lipid peroxidation and hydrogen peroxide generation. Fe-NTA treatment alone caused depletion of glutathione, glutathione metabolizing and antioxidant enzymes in rat liver, whereas pretreatment of animals with vitamin E reversed these changes in a dose-dependent manner. Taken together, our results suggest that vitamin E may afford substantial protection against the damage caused by Fe-NTA exposure and can serve as a potent preventive agent to suppress oxidant-induced tissue injury.     

Molecular properties and enhancement of thermostability by random mutagenesis of glutamate dehydrogenase from Bacillus subtilis

The rocG gene encoding glutamate dehydrogenase from Bacillus subtilis (Bs-GluDH) was cloned, and expressed at considerable magnitude in Escherichia coli. The recombinant Bs-GluDH was purified to homogeneity and has been determined to have a hexameric structure (M(r) 270 kDa) with strict specificity for 2-oxoglutarate and L-glutamate, requiring NADH and NAD+ as cofactors respectively. The enzyme showed low thermostability with T(m) = 41 degrees C due to dissociation of the hexamer. To improve the thermostability of this enzyme, we performed error-prone PCR, introducing random mutagenesis on cloned GluDH. Two single mutant enzymes, Q144R and E27F, were isolated from the final mutant library. Their T(m) values were 61 degrees C and 49 degrees C respectively. Furthermore, Q144R had a remarkably high k(cat) value (435 s(-1)) for amination reaction at 37 degrees C, 1.3 times higher than that of the wild-type. Thus, Q144R can be used as a template gene to modify the substrate specificity of Bs-GluDH for industrial use.     

Interaction of bilirubin with sealed and human serum albumin-entrapped sealed membranes

In order to study the mechanism of entry and localization of bilirubin (BR) into cell membrane, binding of BR to sealed and human serum albumin (HSA)-entrapped sealed membranes was studied by CD spectroscopy. An induced bisignate CD cotton effects (CDCEs) of BR-bound sealed membranes were observed with maxima at 515 nm and minima at 470 nm with a shoulder at 430 nm. BR-bound HSA-entrapped sealed membranes produced CD spectra with additional positive peaks at 450 and 475 nm and negative troughs at 390 and 415 nm. The induced CDCEs of BR-bound sealed membranes and BR-bound HSA-entrapped sealed membranes were perturbed by the addition of drugs (ceftriaxone and sodium salicylate) with the effect of ceftriaxone being more pronounced. Drugs’ being the displacer of BR from albumin, their incorporation in the incubation mixture was paralleled by reduction in CDCEs. Taken together, these results suggest that BR can traverse the membrane bilayer towards the inner surface instead of remaining intercalated in the exterior half of the bilayer.     

Conductivity and pH dual detection of growth profile of healthy and stressed Listeria monocytogenes

In this study, growth of Listeria monocytogenes in a low conductivity growth medium (LCGM) was simultaneously monitored by conductivity and pH measurements. Detection times obtained from the conductivity and pH growth curves were inversely related to the initial concentration of L. monocytogenes in the medium. Linear responses were found by plotting detection times obtained from both conductivity and pH growth curves as a function of initial cell concentration in the range of 10(2) to 10(7) cfu/mL. The detection time was approximately 12 and 2 h for 10(2) and 10(7) cfu/mL of viable L. monocytogenes, respectively, using the conductivity growth curves, whereas it was approximately 1 h less using the pH growth curves. This dual detection system was used for evaluating the growth of acid-, temperature-, and salt-treated L. monocytogenes in the medium. Acid stress at pH 2 and 3 for 3 h caused approximately 12 and 4 h delay in the detection time on pH growth curves, while stress at pH 5 for 3 h did not cause a significant delay in detection time. Delay in detection times was also observed for L. monocytogenes cells exposed to 45 degrees C for more than 1 h (2 and 6 h). Exposure to 10% NaCl for 3 h did not cause visible delay in the detection time. These observations on detection times for stressed L. monocytogenes had a consistent trend with the cell number decrease determined by surface plating method.     

Cholinesterase inhibitory pregnane-type steroidal alkaloids from Sarcococca hookeriana

The bioassay-guided phytochemical investigation on Sarcococca hookeriana have resulted in the isolation of four new pregnane-type steriodal alkaloids: hookerianamide-D [(2'E,20S)-20-(N,N-formyl(methyl)amino)-3beta-(3',4'-dimethyl-2'-pentenamido)-5alpha-pregnane] (1), hookerianamide-E [(2'E,20S)-20-(N,N-dimethylamino)-3beta-(senecioylamino)-5alpha-pregn-14-en-2beta-O-acetate] (2), hookerianamide-F [(2'E,20S)-20-(N-methylamino)-3beta-(tigloylamino)-5alpha-pregn-2,14-dien-4-one] (3), and hookerianamide-G [(20S)-20-(N,N-dimethylamino)-3beta-(N-methylbenzamido)-5alpha-pregn-4beta-O-acetate] (4), along with five known alkaloids 5-9. Their structures were determined by spectroscopic analysis. These steroidal alkaloids and chemically derived derivatives of compound 5 have displayed varying degree of inhibitory activities against acetylcholinesterase and butyrylcholinesterase enzymes in a concentration-dependent fashion, with the IC(50) values ranging from 1.5 to 148.2 and 0.6 to 100.2 microM, respectively.     

Isolation and identification of a plant growth promotive substance from mixture of essential plant oils

The PCS (commercial products by Field Science Co, Japan, used for air fresheners) was analyzed for the presence of bioactive constituents and their role as root growth promoters. Chromatographic separation of the methanolic solution of PCS resulted in the isolation of an promoting active substance, which was identified using GC-mass spectrometry and NMR spectroscopy as 1,2-propanediol (CH₃CH(OH)CH₂OH). Lettuce seedling growth bioassay as test plant revealed that 1,2-propanediol can act as potent root growth promoter; enhancing the growth of lettuce seedling radicle at a concentration 0.01 ppm. The concentration of 1,2-propanediol in PCS mixture was estimated as 4 g/l. These studies suggest that 1,2-propanediol might play an important role in the plant growth promoting activity of PCS.     

Withanolides, a new class of natural cholinesterase inhibitors with calcium antagonistic properties

The withanolides 1-3 and 4-5 isolated from Ajuga bracteosa and Withania somnifera, respectively, inhibited acetylcholinesterase (AChE, EC 3.1.1.7) and butyrylcholinesterase (BChE, EC 3.1.1.8) enzymes in a concentration-dependent fashion with IC50 values ranging between 20.5 and 49,2 microm and 29.0 and 85.2 microm for AChE and BChE, respectively. Lineweaver-Burk as well as Dixon plots and their secondary replots indicated that compounds 1, 3, and 5 are the linear mixed-type inhibitors of AChE, while 2 and 4 are non-competitive inhibitors of AChE with K(i) values ranging between 20.0 and 45.0 microm. All compounds were found to be non-competitive inhibitors of BChE with K(i) values ranging between 27.7 and 90.6 microm. Molecular docking study revealed that all the ligands are completely buried inside the aromatic gorge of AChE, while compounds 1, 3, and 5 extend up to the catalytic triad. A comparison of the docking results showed that all ligands generally adopt the same binding mode and lie parallel to the surface of the gorge. The superposition of the docked structures demonstrated that the non-flexible skeleton of the ligands always penetrates the aromatic gorge through the six-membered ring A, allowing their simultaneous interaction with more than one subsite of the active center. The affinity of ligands with AChE was found to be the cumulative effects of number of hydrophobic contacts and hydrogen bonding. Furthermore, all compounds also displayed dose-dependent (0.005-1.0 mg/mL) spasmolytic and Ca2+ antagonistic potentials in isolated rabbit jejunum preparations, compound 4 being the most active with an ED50 value of 0.09 +/- 0.001 mg/mL and 0.22 +/- 0.01 microg/mL on spontaneous and K+ -induced contractions, respectively. The cholinesterase inhibitory potential along with calcium antagonistic ability and safe profile in human neutrophil viability assay could make compounds 1-5 possible drug candidates for further study to treat Alzheimer's disease and associated problems.     

Kinetics of novel competitive inhibitors of urease enzymes by a focused library of oxadiazoles/thiadiazoles and triazoles

Based on structure of the substrate of urease and for the purpose of designing pharmacophore models for urease inhibitors, which could be effective in physiological and pharmacological studies, a series of twenty-five 1,3,4-diazole-2(3H)-thiones-2(3H)-thiones, 1,3,4-diazoles-2(3H)-thiones, and 1,2,4-tri-3-thiones (OSNs) were designed, synthesized, and evaluated for various kinetic parameters of urease inhibition. OSNs inhibited the activity of urease(s) in a concentration dependent fashion. Dixon as well as Lineweaver-Burk plots and their secondary replots indicated that the nature of inhibition was of pure competitive type for all the 25 compounds. 5-[4-(hydroxy)phenyl]-1,3,4-thiadiazole-2(3H)-thione was found to be the most active one with a Ki value of 2 microM. The Ki values were increased with an increase in substrate concentrations. Apparently, OSNs employ a homologous mechanism of action by exploiting a common transition catalysis state and acting as ligand chelators to form octahedral complexes with the urease enzymes in an orientation-specific mode. The inhibition was slightly potentiated by lower pH and not abolished in the presence of NH2OH (a scavenger of histidine residue). Because of their safe profile in the genotoxic assay, they may be pursued in the near future for human testing