Relationship between climate and growth of two North African varieties of Pinus pinaster Arn.

Planting tree species that are well adapted to local ecological conditions guarantees the success and sustainability of forest restoration. The aim of this study was to investigate the acclimation of two varieties of Pinus pinaster (var. renoui from Tunisia and var. maghrebiana from Morocco), to the ecological conditions of the Kroumirie Mountains in northwest of Tunisia. Tree growth performance (diameter at 1.30 m [DBH], ring widths and total height) and climate–growth responses over the period 1970–2013 were evaluated for two varieties. The trees used in this study were from pine variety and provenance trials growing in common garden in Souiniet (21 trees per variety). Significant difference in height growth rate, DBH and ring widths was found between the two varieties. The Maghrebiana variety had the highest survival and mean radial growth rates. The mean sensitivity to climate was the same in two varieties. A significant negative correlation between May precipitation and radial growth was found for var. maghrebiana. Both varieties showed a significant negative correlation between May and June temperatures and radial growth. January–February temperatures had a positive influence on ring width. The Maghrebiana variety appears well acclimatised so it is expected to ensure more successful restoration of Kroumirie Mountains.     

Does Ramadan fasting affect acylated ghrelin and growth hormone concentrations during short-term maximal exercise in the afternoon?

Ghrelin is an important stomach hormone that influences several metabolic activities and influences growth hormone (GH) release in response to change in energy homeostasis. The aim of this study was to investigate the effect of Ramadan fasting on acylated ghrelin and GH concentrations during a short-term maximal exercise. Eleven male soccer players (age, 22.11 ± 1.3 years; height, 1.76 ± 0.2 m; body mass, 75.3 ± 2.2; mean ± SD) were asked to perform a 30-s Wingate test, during which we recorded the peak (PP) and mean (MP) powers and fatigue index. Measurements were performed in the afternoon at 18:00 h, after three different occasions: (i) one week before Ramadan (BR), (ii) the first week of Ramadan (FWR), and (iii) the fourth week of Ramadan (ER). Blood samples were taken before, immediately and 60 min after the exercise. Our results show that PP and MP were affected by Ramadan fasting with a significant decrease during Ramadan (i.e. FWR and ER) in comparison with ER (p < 0.05). However, no significant difference was observed between BR and FWR (p > 0.05). Also, a significant decrease in body weight and body fat percentage was observed during Ramadan in comparison with ER. From diet record analysis, protein, fat, and carbohydrate decreased significantly during FWR (p < 0.01), often with further decreases by ER. A significant increase in plasma concentrations of ghrelin was observed during the ER (p < 0.001) compared with BR and FWR, and GH levels were significantly higher during ER (p < 0.01) in comparison with BR and FWR. However, the 30-s Wingate tests had no significant effect on plasma concentrations of ghrelin before or during Ramadan. Our study shows that the concentrations of ghrelin and GH change in the last week of Ramadan fasting, though it is unclear if these changes are related to the changes in body composition during Ramadan, or if they are responsible for the change in performance at the short-duration anaerobic test.     

Diurnal variation in stroke parameters and motor organization in front-crawl swimmers

The aim of the present study was to examine the effects of time of day on stroke parameters and motor organization in front-crawl swimmers. In a randomized order, fourteen regional swimmers (age: 18.7 ± 1.6 years) performed maximal front crawls over 12.5 m during two experimental sessions; the morning sessions were conducted between 07:00 and 09:00 h and the evening experiments were conducted between 17:00 and 19:00 h. Stroke parameters (swim velocity, stroke rate [SR], and stroke length), motor organization (arm stroke phases and arm coordination) were calculated from aerial and underwater side-view cameras. Arm coordination was quantified in terms of an index of coordination (Idc). Results showed that oral temperature was significantly higher in the evening 36.8 ± 0.2 °C than in the morning 36.1 ± 0.2 °C (p < 0.001), with a morning–evening difference of ?0.7 ± 0.1 °C. Performance was also higher in the evening (7.4 ± 0.6 s) than in the morning (8.0 ± 0.8 s) (p < 0.001), with a morning–evening difference of 0.55 ± 0.30 s. Likewise, values of swim velocity and SR were higher in the evening than in the morning (p < 0.001) with morning–evening differences of ?0.10 ± 0.04 m s^?1 and ?3.99 ± 2.91 cycles min^?1, respectively. Percentage Idc increased significantly (p < 0.01) between the morning (?5.1 ± 6.5%) and evening (?1.6 ± 7.0%). It is concluded that maximal swimming trials are performed better in the evening than the morning, and that this might be explained by better stroke parameters and motor organization at this time.     

Precise determination of protein antigenic structures has unravelled the molecular immune recognition of proteins and provided a prototype for synthetic mimicking of other protein binding sites

Summary Intensive research in the author's laboratory had culminated in the determination and synthesis of all the antigenic sites of myoglobin in 1975 and of lysozyme in 1978. Very recently most of the antigenic sites of serum albumin were also localized and synthesized. These investigations provided the first unique insight into the molecular features responsible for the immune recognition of protein antigens and of the factors which determine and regulate the antigenicity of the sites. But moreover, these studies have charted a multi-approach chemical strategy for investigation and synthetic duplication of protein binding sites. Furthermore, the concept of surface-simulation synthesis, which we introduced and developed during our determination of the antigenic structure of lysozyme, has provided a remarkable dimension of unlimited versatility for the synthetic mimicking of any type of protein binding sites. In this concept, the spatially adjacent residues of a protein binding site are linked directly via peptide bonds with appropriate spacers into a single peptide which does not exist in the protein but mimicks a surface region of it. This has proved to be a powerful concept in protein molecular recognition and has opened up many untapped avenues in investigation, duplication and perhaps manipulation of a variety of protein activities. In fact, binding sites representing other protein activities (including antibody combining sites) have or are now being mimicked synthetically in our laboratory by the concept of surface-simulation synthesis.     

Amino acid substitutions outside a preselected antigenic region in hemoglobin affect the binding to monoclonal antibodies obtained by immunization with the synthetic region

It is often assumed that amino acid substitutions outside a protein antigenic site have no effect on the reactivity of a protein variant with antibodies, especially monoclonal antibodies (mAbs). Substitutions that exert an effect on the reactivity of a protein variant with mAbs are frequently considered to be within the antigenic site of the mAb. To test this assumption, two mAbs [IgG_l(k) and IgG_2a (k)] were prepared by immunization with a synthetic peptide corresponding to region 63–78 of the chain of human hemoglobin (Hb). The peptide was used as an immunogen in its free form (i.e., without conjugation to a carrier), so that the results will not be made ambiguous by peptide modification nor by an immune response to sites spanning peptide and protein carrier. In addition to their reaction with human Hb, the mAbs were also studied with four primate Hbs which had no substitutions within region 63–78 and only a limited number of substitutions which occurred outside of, and at considerable distances in three-dimensional (3D) structure from, this region. Inhibition studies revealed substantial differences in the binding affinities of some of the primate Hbs, relative to human Hb. Some of the substitutions caused major decreases in binding, although they were at considerable distances in the 3D structure from the indicated site residues. It is concluded that substitutions in a protein, even when distant from an antigenic site, can exert major influences on the protein's reactivity with anti-site mAbs.     

Effects of amino acid substitutions outside an antigenic site on protein binding to monoclonal antibodies of predetermined specificity obtained by peptide immunization: Demonstration with region 15–22 (antigenic site 1) of myoglobin

Monoclonal antibodies (mAbs) of predetermined specificity were prepared by immunizing with a free (i.e., not conjugated to any carrier) synthetic peptide representing region 15–22 (site 1) of sperm whale myoglobin (SpMb). The cross-reactions of Mb variants with three mAbs were studied in order to determine whether such interactions are influenced by substitutions outsde the site. Finback whale Mb, which has no substitutions within region 15–22, showed lower cross-reactivity and relative binding affinity than the reference antigen, SpMb. Bottle-nose Atlantic dolphin myoglobin (BdMb) and badger myoglobin (BgMb), although they have identical substitutions within region 15–22 (Ala-15 to Gly and Val-21 to Leu), showed very different binding properties. The cross-reaction of BdMb was quite comparable to that of SpMb, while that of BgMb was much lower. Since the two proteins have identical structures in regions 15–22, the differences in their cross-reactivities are readily attributed to the effects of substitutions outside this region. Another pair of myoglobins, horse myoglobins (HsMb) and chicken myoglobin (ChMb), also have two identical substitutions (Ala-15 to Gly and Val-21 to Ile) within region 15–22, but possessed different cross-reactivity. The results indicate that the reaction of mAbs, whose specificity is precisely known and predetermined by the immunizing free peptide, can be markedly affected by substitutions outside the indicated binding region on the protein.     

Hemoglobin binding with haptoglobin: Delineation of the haptoglobin binding site on the α-chain of human hemoglobin

Previous studies from this laboratory employing a comprehensive synthetic overlapping peptide strategy showed that the -chain of human hemoglobin (Hb) contains a single haptoglobin (HP) binding region residing within residues 121–135. The present study describes a precise delineation of this Hp-binding site on the -chain. Two overlapping peptides (111–125 and 121–135) spanning this region and a panel of five peptides decreasing at the C-terminal from residue 135 by decrements of two residues (119–135, 119–133, 119–131, 119–129, and 119–127) were synthesized, purified, and characterized. Quantitative radiometric titration of¹²⁵I-labeled human HP (type 2-1) with adsorbents of each of these synthetic peptides showed that the peptide 119–127 retained a Hp-binding activity equivalent to that of peptide 121–135. This finding indicated that Lys-127 marked the C-terminal boundary of the binding site. Another panel of eight peptides was then synthesized, which had their C-terminus fixed at Lys-127 and increased at the N-terminus by one-residue increments from residue 122 up to residue 115 (122–127, 121–127, 120–127, 119–127, 118–127, 117–127, 116–127, and 115–127). The binding of¹²⁵I-Hp to adsorbents of these peptides demonstrated that the N-terminal boundary of the site did not extend beyond Valine 121. It is, therefore, concluded that the Hp-binding site on the -chain of human Hb comprises residues 121–127.     

Synthesis of tolerogenic monomethoxypolyethylene glycol and polyvinyl alcohol conjugates of peptides

Recent studies from this laboratory showed that tolerogenic peptide conjugates are very effective reagents for obtaining epitope-specific immunosuppression of antibody responses to immunopathogenic sites on multideterminant complex protein antigens. This paper describes the procedure for synthesis of well-defined conjugates of peptides to monomethoxypoly-ethylene glycol (mPEG) or to polyvinyl alcohol (PVA). The first step involves succinylation of the hydroxyl groups on the polymers by reaction with succinic anhydride. The polymer is then coupled via the carboxyl of the succinyl group to the -NH₂ of the completed peptide on the synthetic resin, while maintaining intact all the side-chain protecting groups on the peptide. The mPEG or PVA-peptide conjugates are cleaved from the resin and purified by standard procedures. This method results in the preparation of conjugates in which one molecule of tolerogenic polymer is coupled to the N-terminal of an otherwise unaltered peptide molecule.