Malate synthase from Gossypium hirsutum

Malate synthase was purified 2000-fold from cotyledons of dark-germinated cotton, Gossypium hirsutum. The purified enzyme had a pH optimum of 8.2, and an absolute requirement for a divalent cation. Only glyoxylate and acetyl-CoA served as condensation partners. Results obtained with functional-group directed inhibitors suggest the presence of lysine, tyrosine and histidine residues in the active site. Temperature optimum was 40°, and energy of activation was 3.3 kcal/mol. The MW of cotton malate synthase, determined by rate-zonal density gradient sedimentation, was 750 000. Initial-rate studies indicated Michaelis-Menten kinetics. Inhibition by substrate analogs, plus substrate-interaction kinetics gave results consistent with a sequential bireactant mechanism.     

Spectral imaging for precise surgical intervention in Hirschsprung's Disease

We used advanced spectral imaging for intrasurgical decision making in a preclinical study, on a mouse model of Hirschsprung's Disease. Our imaging device sampled areas from normal and abnormal (aganglionic) colon in these animals. Spectral segmentation and classification of the resulting images showed a clear distinction between the normal and aganglionic regions, as confirmed by pathological analysis and use of mutant mice. We developed a simple algorithm that could distinguish normal from aganglionic colon with high spatial resolution and reproducibility, and the following statistics: sensitivity = 97%, specificity = 94%, positive predictive value = 92%, negative predictive value = 98%. These studies showed translational proof of concept that spectral imaging could be used during operations, in real time, to help surgeons precisely distinguish normal from abnormal tissue without requiring traditional biopsy. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

The Concept of Additivity of Drug Combinations in the Case of Quantal Response

The concept of additivity of drug combinations is widely accepted in pharmacology and toxicology. Up to now, no general statistical methods to test that property are available. The present paper gives a mathematical formulation of additivity, a method to fit dose response surfaces under additivity assumption and a statistical test.     

PDB-wide identification of physiological hetero-oligomeric assemblies based on conserved quaternary structure geometry

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Variability of qualitative and quantitative secondary metabolites traits among wild genetic resources of Lavandula stoechas L.

Lavandula stoechas L. (Lamiaceae) is an attractive shrub native to the Mediterranean regions used for ornamental, melliferous, aromatic and medicinal purposes. Furthermore, this species presents an increasing interest in cosmetics, perfumery and pharmaceutical industries. The variability of qualitative and quantitative metabolic traits among nine wild germplasms representing the distribution area of this species in Tunisia was undertaken. A total of 45 essential oil components were identified in the aerial parts of the studied germplasms. The main essential oil components were camphor (15.32–50.63%), fenchone (6.57–34.70%), 1,8-cineole (0.05–13.45%) and γ-gurjunene (1.10–12.15%). In addition to the well known chemotypes camphor/fenchone and camphor/1.8-cineole, a new chemotype camphor/γ-gurjunene was detected in Tunisian L. stoechas L. Six phenolic acids (quinic acid, gallic acid, p-coumaric acid, 4,5-di-O-caffeoyquinic acid, salviolinic acid and trans cinnamic acid) and five flavonoids (luteolin-7-o-glucoside, naringin, apegenin-7-o-glucoside, quercetin and kampherol) were identified in the ethanolic extracts. Salviolinic acid (46.30–615.18 μg/g) and luteolin-7-o-glucoside (5.98–38.54 μg/g) were the most abundant phenolic compounds. A high significant phytochemical variability (p ˂ 0.01) in the accumulation of volatile and phenolic secondary metabolites among the studied germplasms was recorded. The conducted multivariate (PCA) and clustering (HCA) analyses revealed different classification pattern for essential oil and phenolic compounds. The detected phytochemical polymorphism among the investigated lavender ecotypes didn't show accordance with bioclimatic and geographical areas which suggests genetic background as the main explaining factor. The detected secondary metabolites polymorphism valorises Tunisian L. stoechas L. genetic resources as valuable plant material in further breeding programs. Moreover, an urgent in situ and ex situ conservation measures are required for these wild germplasms threatened by human over-harvesting practices and the occurring dramatic changes in climatic conditions.     

Composition of uterine milk and its changes with gestational period in red stingrays (Hemitrygon akajei)

Uterine milk is secreted in the uterus for embryo nutrition in several elasmobranch species and may contribute to rapid embryonic growth, but the details of its composition and its functions are poorly understood. In this study, to explore the roles of uterine milk for embryos, its components throughout the gestational period were analysed in detail. Uterine milk was collected from pregnant red stingrays (Hemitrygon akajei) in the early, middle and late gestational periods, respectively (n= 3 for each period). The crude composition, constituent proteins and fatty acids in the milk were analysed. The uterine milk was rich in proteins throughout the gestational period, whereas lipids dramatically increased in the middle period and reduced slightly towards the late period. Some proteins potentially associated with nutrition, cartilage growth and embryonic immunity were found. Several enzymes related to central metabolism were also detected. The constituent fatty acids in the middle and late periods were similar to those in the egg yolks of elasmobranchs, except for C18:2, which was rich only in the uterine milk. The most abundant fatty acid in the milk was C16:1, which could function as a lipokine to promote lipid metabolism in the embryo. This study's data suggest that uterine milk may be secreted in addition to the egg yolk in elasmobranchs to support rapid and healthy embryonic growth.     

Evaluation of Antioxidant,Antibacterial and Enhancement of Antibiotic Action by Punica granatum Leaves Crude Extract and Enriched Fraction against Multidrug-Resistant Bacteria

The aim of this study was to evaluate the phytochemical composition, antioxidant, and antimicrobial potential of crude extract and fractions of Punica granatum leaves. The extract was produced by turbo extraction, after which hexanic, ethyl acetate, and aqueous fractions were obtained by partitioning. The chemical analyses were performed by thin layer chromatography and high-performance liquid chromatography, and the antioxidant activities were assayed by DPPH^. and ABTS^.+. Minimal inhibitory and bactericidal concentrations (MIC/MBC) were applied to twenty-two bacteria. Most strains susceptible to extract/fractions and resistant to antibiotics were selected, and ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with the ethyl acetate fraction (EAF) against multidrug-resistant strains in modulatory and checkboard models. The data from chromatographic analyses showed flavonoids and tannins in the extract, as well as the enrichment of EAF in phenols, mainly flavonoids. The flavonoids were connected to the electron transfer activity demonstrated in the DPPH^. and ABTS^.+ assays. Gram-positive strains are more susceptible to EAF. The subinhibitory concentrations of P. granatum enhanced the antimicrobial activity of the agents and reduced the EAF individual MIC, and the combination of EAF and antibiotics demonstrated a synergistic effect. These results present a promising approach for developing a therapy in which antioxidant extracts and fractions can be used in combination with antibiotics.