ADRA2A is involved in neuro‐endocrine regulation of bone resorption

Adrenergic stimulation is important for osteoclast differentiation and bone resorption. Previous research shows that this happens through β2‐adrenergic receptor (AR), but there are conflicting evidence on presence and role of α2A‐AR in bone. The aim of this study was to investigate the presence of α2A‐AR and its involvement in neuro‐endocrine signalling of bone remodelling in humans. Real‐time polymerase chain reaction (PCR) and immunohistochemistry were used to investigate α2A‐AR receptor presence and localization in bone cells. Functionality of rs553668 and rs1800544 single nucleotide polymorphism SNPs located in α2A‐AR gene was analysed by qPCR expression on bone samples and luciferase reporter assay in human osteosarcoma HOS cells. Using real‐time PCR, genetic association study between rs553668 A>G and rs1800544 C>G SNPs and major bone markers was performed on 661 Slovenian patients with osteoporosis. α2A‐AR is expressed in osteoblasts and lining cells but not in osteocytes. SNP rs553668 has a significant influence on α2A‐AR mRNA level in human bone samples through the stability of mRNA. α2A‐AR gene locus associates with important bone remodelling markers (BMD, CTX, Cathepsin K and pOC). The results of this study are providing comprehensive new evidence that α2A‐AR is involved in neuro‐endocrine signalling of bone turnover and development of osteoporosis. As shown by our results the neurological signalling is mediated through osteoblasts and result in bone resorption. Genetic study showed association of SNPs in α2A‐AR gene locus with bone remodelling markers, identifying the individuals with higher risk of development of osteoporosis.     

Duetting Behaviour in the Leafhopper Aphrodes makarovi (Hemiptera: Cicadellidae)

Mate recognition and location in Cicadellidae is mediated exclusively via substrate-borne vibrational signals. In the present study we investigated vibrational signals and mate searching behaviour of the leafhopper Aphrodes makarovi. We studied mating behaviour and exchange of vibrational signals between live insects and in playback experiments. Males emitted long and complex calling signals composed of several sections. Female reply was long and always overlapped the end of the male call. The exchange of male and female vibrational signals was a complex and dynamic interaction during which both partners modified their signals according to partner’s reply. The duration of female reply was influenced by the duration of the male call to which she was responding, while the duration of male call was influenced by the duration of the previous female reply. Such relationship suggests the role of sexual selection in the evolution of male vibrational signals.     

Synthesis, structural studies, and cytostatic evaluation of 5,6-di-O-modified L-ascorbic acid derivatives

The 5,6-di-O-tosylated derivative of l-ascorbic acid was synthesized by selective protection and deprotection of 2,3- and 5,6-dihydroxy functional groups involving 5,6-ditosylation in the final step, while the novel 6-acetoxy, 6-hydroxy, and 6-chloro derivatives of 4,5-didehydro-l-ascorbic acid were obtained by reaction of ditosylated compound with nucleophilic reagents. The analysis of 3JH-4-H-5 homonuclear coupling constants shows that all l-ascorbic acid derivatives except for epoxy and 4,5-didehydro compounds exist in high population as gauche conformers across C-4-C-5 bonds, while 3JC-3-H-5 heteronuclear coupling constants in 4,5-didehydro derivatives indicate cis geometry along C-4-C-5 double bond. The X-ray crystal structure analysis of 2,3-di-O-benzyl-5,6-epoxy- and 5,6-isopropylidene-l-ascorbic acid shows that the oxygen atoms attached at positions 2 and 3 of the lactone ring are disposed in a synperiplanar fashion. Besides that, the dioxolane ring adopts half-chair conformation. The molecules of epoxy derivative are joined into infinite chains by one weak hydrogen bond of C-H...O type. Two O-H...O, and C-H...O hydrogen bonds link the molecules of 5,6-di-O-isopropylidene compound into two-dimensional network. 6-Chloro derivative of 2,3-di-O-benzyl-l-ascorbic acid showed the best cytostatic effects against all tested malignant tumor cells (IC50: approximately 18 microM).     

Monitoring of cross-country skiers by means of an expert model of potential performance

On the basis of expert knowledge, an expert model of potential performance covering the motor, morphological, psychological, and sociological subspace was constructed (MMPS). The scores of variables were obtained by application of the computer program Sport Measurement Management System (SMMS). In the subjects included in measurements, trends of the obtained average scores of variables were established through various competition categories and age periods. The sample of subjects consisted of 48 cross-country skiers in three competition categories. Fluctuations in development in individual age periods are larger in the motor and morphological subspace. In the psychological subspace, an upward trend of average scores can be noticed, while the sociological subspace is not subjected to any essential changes in different age and competition categories. Monitoring of cross-country skiers across all three competition categories showed that in these age categories there are periods which owing to laws of development do not allow uniform progress. Therefore, the principle of individuality must be taken into account especially in planning the transformation process.     

Xylophagous entomofauna in branches of oaks (Quercus spp.) and its significance for oak health in the Karst region of Slovenia

Samples of dying branches from 121 trees of Quercus pubescens, Q. cerris, and Q. petraea were taken from 102 sites in Seslerio autumnalis-Quercetum petraeae and Ostryo-Quercetum pubescentis forests. After nine months’ rearing in eclectors, branches were cut to 20-cm sections, and signs of infestation as well as larvae and adult insects were noted. We analysed 395 sections of branches from Q. pubescens, 125 sections from Q. cerris, and 85 sections from Q. petraea. We found 44 adult insects: 45% were from the Cerambycidae family (Callimus angulatus ssp. angulatus, Poecilium alni, and Exocentrus adspersus), 18% were from the Scolytinae subfamily (Scolytus intricatus, Xyleborus dispar), 4% from the Buprestidae family (Coraebus florentinus), 4.5% from the Cleridae family (Tilloidea unifasciata), and 28.5% from other families. In addition, 145 larvae were found: 50% from the Cerambycidae family, 39% from the Buprestidae family, 4% from the Scolytinae subfamily, and 7% from other families. Significant difference in the abundance of adult insects and larvae with regard to the diameter of branch sections and the host species were found. 78% of dying branches showed signs of infestation. Species of the Buprestidae, Scolytinae, and Cerambycidae represent important factors in the decline in oak vitality in the lower Karst region of Slovenia.     

Problems and progresses in speciation of Al in human serum: An overview

Aluminium (Al) is associated with many clinical disorders in renal patients. Al accumulation in brain has also been related to the neurodegenerative processes in Alzheimer’s disease. In order to better understand Al transport in the human body, it is necessary to identify and quantify chemical species in which Al is present in body fluids and tissues. Among a variety of biological samples, Al speciation was the most frequently investigated in human serum. Improvements were made in the development of analytical techniques for the determination of the amount and composition of high molecular mass Al (HMM-Al) and low molecular mass Al (LMM-Al) species in human serum. However, due to the complex chemistry of Al in serum, its low total concentration and the high risk of contamination, speciation of Al in biological samples is still a difficult task for analytical chemists. In this work, problems related to speciation of Al in human serum are critically discussed. An overview of the progress that was made by the use of different analytical procedures, in order to propose analytical protocols for reliable speciation of Al in serum at low ng mL⁻¹ concentration range, is presented.     

NMR structure of the let-7 miRNA interacting with the site LCS1 of lin-41 mRNA from Caenorhabditis elegans

We have determined the 3D structure of a 34-nt RNA construct, herein named LCS1co, which mimics the interaction of let-7 microRNA (miRNA) to one of its complementary binding sites, LCS1, in the 3′-untranslated region of lin-41 mRNA by solution-state NMR spectroscopy. let-7 miRNAs control the timing of development of the nematode Caenorhabditis elegans and are highly conserved in mammals. The sequence and structure of the two conserved let-7 complementary sites, LCS1 and LCS2, in the 3′-untranslated region of lin-41 mRNA are important for a proper downregulation of lin-41. The high-resolution NMR structure reveals details of the binding of let-7 miRNA to lin-41 mRNA which involves formation of a complex with non-canonical structural elements within the seed region. LCS1co exhibits a stem-loop structure with two stems, an asymmetric internal loop and an adenine bulge. Comparison with the NMR solution-state structure of the let-7:lin-41 complex involving the LCS2-binding site shows that conformational freedom of the asymmetric internal loop of LCS1co correlates with a smaller bend between the upper and lower stems in comparison to the well-defined asymmetric loop of LCS2co.     

1,4-diazepine-2,5-dione ring formation during solid phase synthesis of peptides containing aspartic acid beta-benzyl ester.

The Fmoc-based SPPS of H-Xaa-Asp(OBzl)-Yaa-Gly-NH(2) sequences results in side reactions yielding not only aspartimide peptides and piperidide derivatives, but also 1,4-diazepine-2,5-dione-peptides. Evidence is presented to show that the 1,4-diazepine-2,5-dione derivative is formed from the aspartimide peptide. The rate of this ring transformation depends primarily on the tendency to aspartimide and piperidide formation, which is influenced by the nature of the amino acid following the aspartic acid beta-benzyl ester (Xaa). However the bulkiness of the amino acid side chain preceeding the aspartic acid beta-benzyl ester (Yaa) is also important. Under certain conditions the 1,4-diazepine-2,5-dione peptide derivative may even be formed dominantly, which is a highly undesirable side reaction in peptide synthesis, but which provides a new way for the synthesis of diazepine peptide derivatives with targeted biological or pharmacological activity.     

A unified model for the origin of DNA sequence-directed curvature

The fine structure of the DNA double helix and a number of its physical properties depend upon nucleotide sequence. This includes minor groove width, the propensity to undergo the B-form to A-form transition, sequence-directed curvature, and cation localization. Despite the multitude of studies conducted on DNA, it is still difficult to appreciate how these fundamental properties are linked to each other at the level of nucleotide sequence. We demonstrate that several sequence-dependent properties of DNA can be attributed, at least in part, to the sequence-specific localization of cations in the major and minor grooves. We also show that effects of cation localization on DNA structure are easier to understand if we divide all DNA sequences into three principal groups: A-tracts, G-tracts, and generic DNA. The A-tract group of sequences has a peculiar helical structure (i.e., B*-form) with an unusually narrow minor groove and high base-pair propeller twist. Both experimental and theoretical studies have provided evidence that the B*-form helical structure of A-tracts requires cations to be localized in the minor groove. G-tracts, on the other hand, have a propensity to undergo the B-form to A-form transition with increasing ionic strength. This property of G-tracts is directly connected to the observation that cations are preferentially localized in the major groove of G-tract sequences. Generic DNA, which represents the vast majority of DNA sequences, has a more balanced occupation of the major and minor grooves by cations than A-tracts or G-tracts and is thereby stabilized in the canonical B-form helix. Thus, DNA secondary structure can be viewed as a tug of war between the major and minor grooves for cations, with A-tracts and G-tracts each having one groove that dominates the other for cation localization. Finally, the sequence-directed curvature caused by A-tracts and G-tracts can, in both cases, be explained by the cation-dependent mismatch of A-tract and G-tract helical structures with the canonical B-form helix of generic DNA (i.e., a cation-dependent junction model).