Species Differences in the Selective Inhibition of Monoamine Oxidase (1-methyl-2-phenylethyl)hydrazine and its Potentiation by Cyanide

The potentiating effects of cyanide on the inhibition of rat liver mitochondrial monoamine oxidase-A & B and of ox liver mitochondrial MAO-B by pheniprazine [(1-methyl-2-phenylethyl)hydrazine] has been studied. Pheniprazine was shown to behave as a mechanism-based MAO inhibitor. For rat liver MAO-B, the initial non-covalent step was characterized by dissociation constant (K _i) of 2450 nM and the first-order rate constant (k ₊₂) for the covalent adduct formation was 0.16 min⁻¹. As a reversible inhibitor it was selective towards rat liver MAO-A (K _i = 420 nM) but the rate of irreversible inhibition of that enzyme was considerably slower (k ₊₂ = 0.06 min⁻¹). MAO-B from ox liver more closely resembled MAO-A from the rat in sensitivity to reversible inhibition by pheniprazine (K _i = 450 nm) but it was closer to rat liver MAO-B in rate of irreversible inhibition (k ₊₂ = 0.29 min⁻¹). The K _i values were significantly decreased in the presence of KCN but there was little effect on the k ₊₂ values. However, sensitivities of the different enzymes to KCN varied widely and considerably higher concentrations of KCN were required for this effect to be apparent with the rat liver mitochondrial MAO-A than with MAO-B from rat and ox liver. The kinetic behaviour of cyanide activation was consistent with partial (non-essential) competitive activation in all cases. Special issue dedicated to Dr. Moussa Youdim.     

Estimation of linear and mass attenuation coefficients of soy–lignin bonded Rhizophora spp. particleboard as a potential phantom material using caesium-137 and cobalt-60

In this study, linear and mass attenuation coefficients of fabricated particleboards intended for use as phantom material were estimated using ¹³⁷Cs and ⁶⁰Co radiation sources. Particleboards made of Rhizophora spp. wood trunk bonded with soy flour and lignin were fabricated at a target density of 1.0 g cm^?3, with and without gloss finish coating. Elemental composition of the particleboards was obtained by means of energy dispersive X-ray (EDX) spectroscopy. Experimental setups were simulated via the GATE Monte Carlo (MC) package, with particle histories of 1?×?10⁶–1?×?10⁷. Linear and mass attenuation coefficients obtained from measurements and GATE simulations were compared and discussed. The percentage differences between the measured and simulated linear and mass attenuation coefficients of the samples were reasonably small (2.05–4.88% for ¹³⁷Cs and 3.24–5.38% for ⁶⁰Co). It is shown that all the particleboards have the potential to be used as phantom materials as the attenuation coefficients measured were in good agreement with those of water (calculated with XCOM) and with those simulated with the GATE toolkit. The use of gloss finish coating also did not show any significant effect on the attenuation coefficient of the phantom material. Verification of experimental results via GATE simulations has been shown crucial in providing reliable data for energy transmission studies. Based on the results achieved in this study, it is concluded that the studied material—Rhizophora spp. wood trunk bonded with soy flour and lignin including gloss finish coating—can be used in radiation dosimetry studies.

     

Dynamic Motions of the HIV-1 Frameshift Site RNA

The HIV-1 frameshift site (FS) plays a critical role in viral replication. During translation, the HIV-1 FS transitions from a 3-helix to a 2-helix junction RNA secondary structure. The 2-helix junction structure contains a GGA bulge, and purine-rich bulges are common motifs in RNA secondary structure. Here, we investigate the dynamics of the HIV-1 FS 2-helix junction RNA. Interhelical motions were studied under different ionic conditions using NMR order tensor analysis of residual dipolar couplings. In 150 mM potassium, the RNA adopts a 43°(±4°) interhelical bend angle (β) and displays large amplitude, anisotropic interhelical motions characterized by a 0.52(±0.04) internal generalized degree of order (GDO_int) and distinct order tensor asymmetries for its two helices (η = 0.26(±0.04) and 0.5(±0.1)). These motions are effectively quenched by addition of 2 mM magnesium (GDO_int = 0.87(±0.06)), which promotes a near-coaxial conformation (β = 15°(±6°)) of the two helices. Base stacking in the bulge was investigated using the fluorescent purine analog 2-aminopurine. These results indicate that magnesium stabilizes extrahelical conformations of the bulge nucleotides, thereby promoting coaxial stacking of helices. These results are highly similar to previous studies of the HIV transactivation response RNA, despite a complete lack of sequence similarity between the two RNAs. Thus, the conformational space of these RNAs is largely determined by the topology of their interhelical junctions.     

Ethanesulfonic acid-based esterification of industrial acidic crude palm oil for biodiesel production

An industrial grade acidic crude palm oil (ACPO) pre-treatment process was carried out using ethanesulfonic acid (ESA) as a catalyst in the esterification reaction. ESA was used in different dosages to reduce free fatty acid (FFA) to a minimum level for the second stage of biodiesel production via alkaline transesterification reaction. Different process operating conditions were optimized such as ESA dosage (0.25-3.5% wt/wt), methanol to ACPO molar ratio (1:1-20:1), reaction temperature (40-70 °C), and reaction time (3-150 min). This study revealed the potential use of abundant quantities of ACPO from oil palm mills for biodiesel production. The lab scale results showed the effectiveness of the pre-treatment process using ESA catalyst. Three consecutive catalyst recycling runs were achieved without significant degradation in its performance. Second and third reuse runs needed more reaction time to achieve the target level of FFA content. Esterification and transesterification using ESA and KOH respectively is proposed for biodiesel industrial scale production. The produced biodiesel meets the international standards specifications for biodiesel fuel (EN 14214 and ASTM D6751).     

Host range of Secusio extensa (Lepidoptera: Arctiidae), and potential for biological control of Senecio madagascariensis (Asteraceae)

Secusio extensa (Lepidoptera: Arctiidae) was evaluated as a potential biological control agent for Madagascar fireweed, Senecio madagascariensis (Asteraceae), which has invaded over 400 000 acres of rangeland in the Hawaiian Islands and is toxic to cattle and horses. The moth was introduced from southeastern Madagascar into containment facilities in Hawaii, and host specificity tests were conducted on 71 endemic and naturalized species (52 genera) in 12 tribes of Asteraceae and 17 species of non‐Asteraceae including six native shrubs and trees considered key components of Hawaiian ecosystems. No‐choice feeding tests indicated that plant species of the tribe Senecioneae were suitable hosts with first instars completing development to adult stage on S. madagascariensis (78.3%), Delairea odorata (66.1%), Senecio vulgaris (57.1%), Crassocephalum crepidioides (41.2%), and at significantly lower rates on Emilia fosbergii (1.8%) and Erechtites hieracifolia (1.3%). A low rate of complete larval development also was observed on sunflower, Helianthus annuus (11.6%), in the tribe Heliantheae. However, sunflower was rejected as a potential host in larval‐feeding and adult oviposition choice tests involving the primary host S. madagascariensis as control. Although larvae died as first instars on most test species, incomplete development and low levels of feeding were observed on nine species in the tribes Heliantheae, Cardueae and Lactuceae. Larvae did not feed on any non‐Asteraceae tested, including species with similar pyrrolizidene alkaloid chemistry, crops, and six ecologically prominent native species. Because all species of Senecioneae are non‐native and weedy in Hawaii, these results indicate that S. extensa is sufficiently host‐specific for introduction for biological control. High levels of feeding damage observed on potted plants indicate that S. extensa can severely impact the target fireweed as well as D. odorata, a noxious weed in native Hawaiian forests.     

Human carotid lesion linoleic acid hydroperoxide inhibits paraoxonase 1 (PON1) activity via reaction with PON1 free sulfhydryl cysteine 284

Paraoxonase 1 (PON1) is an HDL-associated lactonase with antiatherogenic properties. These include dampening the oxidation properties of human carotid lesion lipid extract (LLE), which in turn inactivates the enzyme. The aims of this study were to identify the PON1 inhibitor in LLE and explore the mechanism of inhibition. LLE inhibited both recombinant PON1 and HDL-PON1 lactonase activity in a dose- and time-dependent manner. Addition of antioxidants or electrophiles to LLE did not prevent PON1 inhibition. LLE was unable to inhibit a PON1 mutant lacking Cys284, whereas it did inhibit all other PON1 mutants tested. The inhibitor in the LLE was identified as linoleic acid hydroperoxide (LA-OOH) and inhibition was specific to this hydroperoxide. During its inhibition, PON1 acted like a peroxidase enzyme, reducing LA-OOH to LA-hydroxide via its Cys284. A similar reaction occurred with external thiols, such as DDT or cysteine, which also prevented PON1 inhibition and restored enzyme activity after inhibition. Thus, the antiatherogenic properties of HDL could be, at least in part, related to the sulfhydryl-reducing characteristics of its associated PON1, which are further protected and recycled by the sulfhydryl amino acid cysteine.     

Vitamin B6s inhibit oxidative stress caused by Alzheimer's disease-related Cu(II)-β-amyloid complexes-cooperative action of phospho-moiety

Cu(II) complexes of Alzheimer's disease-related β-amyloid (Aβ) peptides exhibit metal-centered oxidation chemistry. The metallo-Aβ complexes are the hallmark of the disease and have been attributed to the generation of reactive oxygen species (ROS), causing oxidative stress. In this communication, the inhibitions of the oxidative activity of Cu(II)-Aβ by vitamin B6 compounds pyridoxamine (PM), pyridoxine (PN), pyridoxal (PL), and pyridoxal-5'-phosphate (PLP) are presented. These B6's are competitive inhibitors toward dopamine oxidation by Cu(II)-Aβ(1-20), with K(i) values of 1.4, 8.3, 1.2, and 0.2mM, respectively. The phospho-moiety in PLP seems to exhibit cooperative inhibition, affording a clue for future design of inhibitors.     

Discovery of selective bioactive small molecules by targeting an RNA dynamic ensemble

Current approaches used to identify protein-binding small molecules are not suited for identifying small molecules that can bind emerging RNA drug targets. By docking small molecules onto an RNA dynamic ensemble constructed by combining NMR spectroscopy and computational molecular dynamics, we virtually screened small molecules that target the entire structure landscape of the transactivation response element (TAR) from HIV type 1 (HIV-1). We quantitatively predict binding energies for small molecules that bind different RNA conformations and report the de novo discovery of six compounds that bind TAR with high affinity and inhibit its interaction with a Tat peptide in vitro (K(i) values of 710 nM-169 μM). One compound binds HIV-1 TAR with marked selectivity and inhibits Tat-mediated activation of the HIV-1 long terminal repeat by 81% in T-cell lines and HIV replication in an HIV-1 indicator cell line (IC(50) ～23.1 μM).     

Functional morphology of the metapleural gland in workers of the ant Crematogaster inflata (Hymenoptera,Formicidae)

Abstract. Workers of Crematogaster inflata possess the largest metapleural glands (relative to body size) known among ants, with reservoirs extending anteriorly up to the junction between the pro‐ and the mesothorax, and with over 1400 secretory cells on both sides together. This large secretory capacity is related to the gland's defensive function, which, in members of this species, is directed against larger arthropod and vertebrate enemies, and apparently not against microorganisms, in contrast to other ants, where the gland produces antibiotics. The gland is not equipped with any direct musculature. Secretion release is probably caused by contraction of the oblique longitudinal thorax muscles or by passive expulsion caused by external pressure.