Synthesis and biological activity of oxytocin analogues containing unnatural amino acids in position 9: structure activity study

We report the solid phase synthesis and some pharmacological properties of 24 oxytocin (OT) analogues. Basic modifications at position 9 (introduction of l- or d-β-(2-thienyl)-alanine [L- or D-Thi], or l- or d-3-Pyridylalanine [l- or d-3-Pal]) were combined with d-tyrosine(OEthyl) [d-Tyr(Et)] or d-1-naphthylalanine [d-1-Nal] in position 2 and β-mercaptopropionic acid (Mpa) in position 1 modifications in altogether 14 analogues. Additionally, 8 analogues having α-aminoisobutyric acid [Aib] or d-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (d-Tic) or diethylglycine (Deg) in position 9 and d-Tyr(Et) or d-1-Nal or d-Tic in position 2 and Mpa or Pen (ββ-dimethylcysteine) in position 1 were prepared. Two of these analogues have one more modification in position 6, i.e. Pen. Furthermore, two analogues having Mpa in position 1 and d-Tyr(Et) or d-1-Nal in position 2 were prepared for comparison purposes. The analogues were tested for rat uterotonic activity in vitro, in the rat pressor assay and for binding affinity to human OT receptor. The analogue having the highest anti-oxytocic activity was [Mpa¹, d-Tyr(Et)², Deg⁹]OT (pA₂ = 8.68 ± 0.26); this analogue was also selective.     

Melectin MAPs: the influence of dendrimerization on antimicrobial and hemolytic activity

The recently described antimicrobial peptide melectin (MEP, GFLSILKKVLPKVMAHMK-NH₂) exhibits high antimicrobial activity against Gram-positive and Gram-negative bacteria. Here we describe the synthesis and biological activities of 23 new analogues of MEP. We studied the influence of dimerization and tetramerization (MAP-constructs of MEP) on the antimicrobial and hemolytic activities, as well as the role of Met in positions 14 and 17 of the peptide chain. Oxidation of the Met to Met(O) and Met(O₂) decreases antimicrobial activity of all tested bacteria if the peptide is in the monomeric form, however, only to Staphylococcus aureus if in the form of dimer or tetramer. Dimerization and tetramerization increase the undesirable hemolytic activity of the peptides. Interestingly, substitution of Leu for Val in position 6 leads to the decrease of hemolytic activity. Introduction of the isosteric amino acid Nle into positions 14 or 17 or both leads to slight increase of hemolytic activity under preservation of high antimicrobial activities. Unfortunately, dimerization again leads to an increase of hemolytic activity.     

Novel antimicrobial peptides from the venom of the eusocial bee Halictus sexcinctus (Hymenoptera: Halictidae) and their analogs

Two novel antimicrobial peptides, named halictines, were isolated from the venom of the eusocial bee Halictus sexcinctus. Their primary sequences were established by ESI-QTOF mass spectrometry, Edman degradation and enzymatic digestion as Gly-Met-Trp-Ser-Lys-Ile-Leu-Gly-His-Leu-Ile-Arg-NH₂ (HAL-1), and Gly-Lys-Trp-Met-Ser-Leu-Leu-Lys–His-Ile-Leu-Lys-NH₂ (HAL-2). Both peptides exhibited potent antimicrobial activity against Gram-positive and Gram-negative bacteria but also noticeable hemolytic activity. The CD spectra of HAL-1 and HAL-2 measured in the presence of trifluoroethanol or SDS showed ability to form an amphipathic α-helical secondary structure in an anisotropic environment such as bacterial cell membrane. NMR spectra of HAL-1 and HAL-2 measured in trifluoroethanol/water confirmed formation of helical conformation in both peptides with a slightly higher helical propensity in HAL-1. Altogether, we prepared 51 of HAL-1 and HAL-2 analogs to study the effect of such structural parameters as cationicity, hydrophobicity, α-helicity, amphipathicity, and truncation on antimicrobial and hemolytic activities. The potentially most promising analogs in both series are those with increased net positive charge, in which the suitable amino acid residues were replaced by Lys. This improvement basically relates to the increase of antimicrobial activity against pathogenic Pseudomonas aeruginosa and to the mitigation of hemolytic activity.     

Grass cover on forest clear-cut areas ameliorates some soil chemical properties

Clear-cut areas formed after forest decline due to acid deposition, pest attacks, or wind-breaks in temperate mountainous regions are often populated by grass (mainly Calamagrostis villosa). This study focused on the changes of soil chemical characteristics under the grass cover replacing the forest, focusing mainly on aluminium (Al) speciation. Clear-cut area due to strong acid deposition in the Jizera Mountains (Northern Bohemia) was studied. The soils under grass cover exhibit higher pH values and lower exchangeable Al content compared to adjacent surviving forest. Mobile Al species under the grass have larger proportion of non-toxic organic complexes. The content of exchangeable base cations is slightly higher under the grass. The positive effect of grass on soil chemistry was enhanced by liming. The temporary grass cover can therefore improve soil chemical quality for following reforestation. However, the differences are generally limited to surface organic horizons. Similar results were found also on a bark-beetle clear-cut area in the Bohemian Forest (Southern Bohemia) with smaller acid deposition; nevertheless, most differences were not significant there.     

Spatial variation of deuterium enrichment in bulk water of snowgum leaves

Deuterium enrichment of bulk water was measured and modeled in snowgum (Eucalyptus pauciflora Sieber ex Sprengel) leaves grown under contrasting air and soil humidity in arid and wet conditions in a glasshouse. A map of the enrichment was constructed with a resolution of 4 mm by using a newly designed cryodistillation method. There was progressively increasing enrichment in both longitudinal (along the leaf midrib) and transversal (perpendicular to the midrib) directions, most pronounced in the arid-grown leaf. The whole-leaf average of the enrichment was well below the value estimated by the Craig-Gordon model. The discrepancy between model and measurements persisted when the estimates were carried out separately for the leaf base and tip, which differed in temperature and stomatal conductance. The discrepancy was proportional to the transpiration rate, indicating the significance of diffusion-advection interplay (Péclet effect) of deuterium-containing water molecules in small veins close to the evaporating sites in the leaf. Combined Craig-Gordon and desert-river models, with or without the Péclet number, P, were used for predicting the leaf longitudinal enrichment. The predictions without P overestimated the measured values of deltadeuterium. Fixed P value partially improved the coincidence. We suggest that P should vary along the leaf length l to reconcile the modeled data with observations of longitudinal enrichment. Local values of P, P(l), integrating the upstream fraction of water used or the leaf area, substantially improved the model predictions.     

Biomimetic apatite coatings--carbonate substitution and preferred growth orientation

Biomimetic apatite coatings were obtained by soaking chemically treated titanium in SBF with different HCO(3)(-) concentration. XRD, FTIR and Raman analyses were used to characterize phase composition and degree of carbonate substitution. The microstructure, elemental composition and preferred alignment of biomimetically precipitated crystallites were characterized by cross-sectional TEM analyses. According to XRD, the phase composition of precipitated coatings on chemically pre-treated titanium after exposure to SBF was identified as hydroxy carbonated apatite (HCA). A preferred c-axis orientation of the deposited crystals can be supposed due to the high relative peak intensities of the (002) diffraction line at 2theta=26 degrees compared to the 100% intensity peak of the (211) plane at 2theta=32 degrees . The crystallite size in direction of the c-axis of HCA decreased from 26 nm in SBF5 with a HCO(3)(-) concentration of 5 mmol/l to 19 nm in SBF27 with a HCO(3)(-) concentration of 27 mmol/l. Cross-sectional TEM analyses revealed that all distances correspond exactly to the hexagonal structure of hydroxyapatite. The HCO(3)(-) content in SBF also influences the composition of precipitated calcium phosphates. Biomimetic apatites were shown to have a general formula of Ca(10-x-y)Mg(y)(HPO(4))(x-z)(CO(3))(z)(PO(4))(6-x)(OH)(2-x-w)(CO(3))(w/2). According to FTIR and Raman analyses, it can be supposed that as long as the HCO(3)(-) concentration in the testing solutions is below 20 mmol/l, only B-type HCA (0相似文献