Evaluation of the metal-ion-coordinating differences between the 2'-, 3'- and 5'-monophosphates of adenosine

The stability constants of the 1:1 complexes formed between Mg2+, Ca2+, Sr2+, Ba2+, Mn2+, Co2+, Ni2+, Cu2+, Zn2+ or Cd2+ and 2'AMP2-, 3'AMP2- or 5'AMP2- were determined by potentiometric pH titration in aqueous solution (I = 0.1 M, NaNO3; 25 degrees C). The experimental conditions were carefully selected such that self-association of the nucleotides and their complexes is negligibly small; i.e. it was made certain that the properties of the monomeric divalent-metal-ion--AMP [M(AMP)] complexes were studied. Based on recent measurements with simple phosphate monoesters, R-MP2- where R is a non-coordinating residue [Massoud, S. S. & Sigel, H. (1988) Inorg. Chem. 27, 1447-1453], it is shown that all the M(AMP) complexes of the alkaline earth ions, with the possible exception of Mg(5'AMP), have exactly the stability expected for a sole-phosphate coordination of the metal ion. The same property is revealed for the complexes with Mn2+, Co2+, Zn2+ or Cd2+ and 3'AMP2-; in case of Ni(3'AMP) and Cu(3'AMP) a slight stability increase just at the edge of the experimental-error limits is indicated. This slight stability increase is attributed to the formation of a macrochelate (possibly with N-3); in fact, additional information confirms macrochelation for Cu(3'AMP). About 45% of Cu(2'AMP) exists in aqueous solution as a macrochelate (probably involving N-3); the other M(2'AMP) complexes (M2+ = Mn2+, Co2+, Ni2+, Zn2+, Cd2+) form (if at all) only traces of a base-backbound species. Most pronounced is macrochelate formation with 5'AMP2-: all mentioned 3d ions and Zn2+ or Cd2+ form to some extent macrochelates via N-7 (the structures of these closed species are indicated). In case of M(5'AMP) the base-binding site is certain: replacement of N-7 by a CH unit (tubercidin 5'-monophosphate) eliminates any increased complex stability, whereas formation of the 1,N6-etheno bridge to form 1,N6-ethenoadenosine 5'-monophosphate results in the phenanthroline-like N-6,N-7 site which facilitates macrochelation significantly.     

Macro- and microscopic morphology of the flank scales of families Lutjanidae and Serranidae from the Persian Gulf Coral Reefs (Teleosts: Perciformes)

Macro- and microscopic characteristics of flank scales for 12 species were investigated from the Persian Gulf Coral Reefs. In Lutjanus argentimaculatus and L. russellii (family Lutjanidae), the scales of different flank regions were not different, while four characters showed variation in the scale of L. lutjanus i.e., scale shape (pentagonal, hexagonal and square), anterior margin (waved, scalloped and smooth), focus shape (circular and oblong) and focus position (postero-central and central), displayed variation. Scale type (ctenoid) and posterior margin (transforming ctenii) did not show variation and could be considered to be specific in this family. In Epinephelus chlorostigma (family Serranidae), the scales of flank regions did not display variation, while in E. areolatus, E. diacanthus and E. radiates, the scales showed considerable variation. The most variable characters were scale shape, posterior margin and focus shape. Therefore, in fish systematics studies on the base of scale, it is particularly important to compare scales from the same flank regions. Also, some criteria such as size-dependent alternation, ontogenetic changes and variation between flank regions, should be considered. This study supports the potential of scale morphology to help for the understanding of fish diversity in the coral reef ecosystem.     

Genetic relationships among actinomycetes that produce the immunosuppressant macrolides FK506, FK520/FK523 and rapamycin

Summary A polyphasic taxonomic study was undertaken to establish the genetic and phenotypic relationships among six actinomycetes that produce the immunosuppressant macrolides FK506, FK520/FK523 and rapamycin. Chemotaxonomic studies reveal that all have Type I cell walls. Gas chromatography (GC) of fatty acid methyl esters revealed patterns consistent for strains ofStreptomyces with 160 and 150anteiso predominating. Principal component analysis of GC data revealed distinct profiles for each culture. Reciprocal DNA homology studies atT _m -25 showed the rapamycin-producing strain and one FK506-producing strain to have 38–50% homology with the type strain ofStreptomyces hygroscopicus (ATCC 27438). The remaining strains exhibited 6–17% homology. To further explore the relationships among these strains all were probed for the presence of anO-methyltransferase gene specific to this biosynthetic pathway. Among the strains of interest, onlyStreptomyces hygroscopicus subsp.yakushimaensis, the patent strain for FK520/FK523, failed to hybridize with the probes.     

Exosomes for angiogenesis induction in ischemic disorders

Ischaemic disorders are leading causes of morbidity and mortality worldwide. While the current therapeutic approaches have improved life expectancy and quality of life, they are unable to “cure” ischemic diseases and instate regeneration of damaged tissues. Exosomes are a class of extracellular vesicles with an average size of 100–150 nm, secreted by many cell types and considered a potent factor of cells for paracrine effects. Since exosomes contain multiple bioactive components such as growth factors, molecular intermediates of different intracellular pathways, microRNAs and nucleic acids, they are considered as cell-free therapeutics. Besides, exosomes do not rise cell therapy concerns such as teratoma formation, alloreactivity and thrombotic events. In addition, exosomes are stored and utilized more convenient. Interestingly, exosomes could be an ideal complementary therapeutic tool for ischemic disorders. In this review, we discussed therapeutic functions of exosomes in ischemic disorders including angiogenesis induction through various mechanisms with specific attention to vascular endothelial growth factor pathway. Furthermore, different delivery routes of exosomes and different modification strategies including cell preconditioning, gene modification and bioconjugation, were highlighted. Finally, pre-clinical and clinical investigations in which exosomes were used were discussed.     

Synthesis,anticancer activity,and β‐lactoglobulin binding interactions of multitargeted kinase inhibitor sorafenib tosylate (SORt) using spectroscopic and molecular modelling approaches

Sorafenib tosylate (SORt) is an oral multikinase inhibitor used for treatment of advanced renal cell, liver, and thyroid cancers. In this study, this drug was synthesized and its antiproliferative activities against HCT116 and CT26 cells were assessed. The interaction of SORt with β‐lactoglobulin (BLG) was studied using different fluorescence techniques, circular dichroism (CD), zeta potential measurements, and docking simulation. The results of infrared (IR), mass, ^HNMR, and ^CNMR spectra demonstrated that the drug was produced with high quality, purity, and efficiency. SORt showed potent cytotoxicity against HCT116 and CT26 cells with IC₅₀ of 8.12 and 5.42 μM, respectively. For BLG binding of SORt, the results showed that static quenching was the cause of the high affinity drug–protein interaction. Three‐dimensional fluorescence and synchronous spectra indicated that SORt conformation was changed at different levels. CD suggested that the α‐helix content remained almost constant in the BLG–SORt complex, whereas random coil content decreased. Zeta potential values of BLG were more positive after binding with SORt, due to electrostatic interactions between BLG and SORt. Thermodynamic parameters confirmed van der Waals and hydrogen bond interactions in the complex formation. Molecular modelling predicted the presence of hydrogen bonds and electrostatic forces in the BLG–SORt system, which was consistent with the experimental results.     

Optimization of the Emerson–Trinder enzymatic reaction by response surface methodology

The Emerson–Trinder reaction has been optimized in this work using an initial rate spectrophotometric method and response surface methodology (RSM). In this investigation, the variation range of critical variables along with the fixed parameters were selected based on a preliminary ‘one at a time’ (OVAT) procedure for the subsequent RSM chemometric analysis as follows: pH (6–10), buffer concentration (50–250?mM), 4-aminoantipyrine (4-AAP) concentration (1–5?mM), temperature (25–45°C). The optimum values of fixed parameters were: 4-fluorophenol (4-FP, 30?mM), horseradish peroxidase (HRP) enzyme activity (0.12?U?mL^?1), and the fixed concentration of the H₂O₂ in the chemometric experiments was 11.4 µM. The non-linear nature of the experimental response of the reaction system was explained by a second-order polynomial equation, which revealed the impact of the experimental factors, their interactions and also their optimum values. The results of the reported RSM analysis proved to be quite appropriate for the design and optimization of this reaction, as illustrated by the relatively high value of the determination coefficient (R²=96.7%) for the fitting of quadratic model, along with the satisfactory results generated by the analysis of variance (ANOVA). All the evaluated analytical characteristics of this method: typical reaction progress curves, resulting linear calibration curve, within-day precisions at low and at high levels, and the upper and lower detection limits were, also, reported. In addition, to check the quality of the optimization and validity of the model, the assay of H₂O₂, in pooled serum matrix and in cosmetic samples, was performed.     

Centaurea shahuensis and C. ravansarensis spp. nov. (Asteraceae,Cardueae) from west Iran

Two new species of Centaurea L. sect. Cynaroides Boiss. ex Walp. (Asteraceae), C. shahuensis Ranjbar and Negaresh and C. ravansarensis Ranjbar and Negaresh are described and illustrated from Kermanshah Province, west Iran. They are closely related to C. regia Boiss. subsp. regia. However, C. shahuensis differs from it by median stem leaves broadly oblanceolate or subpandurate, phyllaries densely lanate‐tomentose, appendages small, concealing a minor part of phyllaries, and also median appendage margin entire sometimes with 1–2 cilia, 1.2–3.0 mm long on each side. Centaurea ravansarensis is distinguished by upper stem leaves loosely arachnoid, phyllaries loosely floccose‐tomentose, inner appendages deep brown to blackish, and spine 4.5–6.0 mm long. Habitat, conservation status and the geographical distribution of the new species are given.     

Liver alteration and hematological and serum biochemical responses of common carp,Cyprinus carpio Linnaeus, 1758, following long‐term feeding of pistachio (Pistacia vera) green hull extract as a source of natural phenol

This study was conducted to investigate the effect of pistachio green hull extract (PGHE) on hematological and serum biochemical changes in common carp, Cyprinus carpio. Three hundred common carp (11.65 ± 1.65 g) were fed one of five different dietary treatments (with three replications) containing 0, 0.5, 1.5, 4.5 or 9 g PGHE kg^?1diet for ten continuous weeks. Each tank had a 90‐L capacity and water flow rate of about 500 ml min^_1.Total phenolic compounds of the different diets differed significantly (P < 0.001) according to the amount of PGHE. At the end of experiment, six fish were removed randomly from each treatment. Blood samples were taken for hematological and serum biochemical analyses at room temperature. Liver tissue samples were processed for histology and stained by H&E. The results indicated that all doses of tested PGHE induced no significant changes in hematocrit, hemoglobin, or erythrocytes, nor alkaline phosphatase, alanine transaminase, lactate dehydrogenase, total protein, albumin, globulin, triglycerides, low‐density lipoprotein, high‐density lipoprotein, glucose or cholesterol in the serum. Leukocytes were higher (P < 0.01) in fish fed a 1.5, 4.5, or 9 g PGHE kg^?1 diet when compared to the 0.5 g PGHE kg^?1 diet group or the control. Serum aspartate transaminase in treatments containing a 4.5 or 9 g PGHE kg^?1 diet was significantly (P < 0.001) higher in comparison with the control. Liver histology showed focal necrosis, cytoplasm degeneration, lateral nuclei and an increase in Kupffer cells following PGHE administrations. The results of this trial indicated that although there were no significant changes in most hematological and biochemical parameters, PGHE could induce some adverse pathological effects on liver tissue.     

Differentiation and Classification of Phytoplasmas Associated with Almond and GF‐677 Witches'‐Broom Diseases using rRNA and rp Genes

Stone fruits are affected by several diseases associated with plant pathogenic phytoplasmas. Previous studies have been shown that phytoplasma agents of almond and GF‐677 witches'‐broom (AlmWB and GWB, respectively) diseases belong to pigeon pea witches'‐broom (16SrIX) phytoplasma group. In this study, partial biological and molecular characterization was used to compare and classify phytoplasma agents of Khafr AlmWB (KAlmWB) and Estahban GWB (EGWB) diseases. Production of different symptoms in periwinkle indicated that agents of KAlmWB and EGWB are differentiable. Expected fragments were amplified from diseased almond and GF‐677 trees in direct PCR using phytoplasma universal primer pairs P1/P7 and rpF1/rpR1 and nested PCR using P1/P7 followed by R16F2n/ R16R2 primer pair. 16S‐rDNA Restriction fragment length polymorphism (RFLP) as well as phylogenetic analysis of rplV‐rpsC and 16S–23S rRNA spacer region sequences classified KAlmWB and EGWB phytoplasmas within 16SrIX‐C (rpIX‐C) and 16SrIX‐B (rpIX‐B) subgroups, respectively.     

The influential factor of narcolepsy on quality of life: compared to obstructive sleep apnea with somnolence or insomnia

Sleep and Biological Rhythms - Narcoleptics tend to have a low quality of life (QoL). Few studies have compared QoL in narcolepsy against other sleep disorders. The purpose of this study was to...