Effects of forest thinning on sap flow dynamics and transpiration in a Japanese cedar forest.

Historically, forest thinning in Japan was conducted to obtain high-quality timber from plantations. Today, in contrast, thinning is also motivated by forest water balance and climate change considerations. It is in this context that the present study examines the effects of thinning on the ecophysiological responses of remaining trees, which are inadequately understood, especially in relation to changes in the magnitude and duration of transpiration. Sap flux densities were measured in both outer and inner sapwood to obtain stand-scale transpiration for two years in the pre-thinning state and three years post-thinning. The effects of thinning on transpiration were quantitatively evaluated based on canopy conductance models. The larger increases in outer sap flux density were found in the first year after the treatment, while those in inner sap flux density were detected in the second and third years. The remaining trees required a few of years to adjust to improved light conditions of the lower crown, resulting in a delayed response of inner sap flux density. As a result of this lag, transpiration was reduced to 71 % of the pre-thinning condition in the first year, but transpiration recovered to the pre-thinning levels in the second and third years due to compensating contributions from inner sap flow. In terms of more accurately chronicling the thinning effect, the distribution of sap flux density with respect to its radial pattern, is necessary. Such measurements are key to more comprehensively examining the ecophysiological response of forest plantations to thinning and, ultimately, its effect on the forest water balance.

Multisite Partial Glycosylation Approach for Preparation of Biologically Relevant Oligomannosyl Branches Contribute to Lectin Affinity Analysis.

High-mannose-type glycans play essential biological roles, e.g., immune response and glycoprotein quality control, and preparing a series of oligomannosyl branches of high-mannose-type glycans is critical for biological studies. However, obtaining sufficient amounts of the various oligomannosyl branches is challenging. In this study, we demonstrated a partial glycosylation strategy for the single-step synthesis of various biologically relevant oligomannosyl-branched structures. First, Manα1-6(Manα1-3)Man-type oligomannosyl branch was synthesized via double glycosylation from a 3,6-di-OH mannosyl acceptor and fluorinated mannosyl donor with perfect α-selectivity. Subsequent partial glycosylation by reducing the equivalent of the mannosyl donor enabled to obtain biologically relevant Manα1-2Manα1-6(Manα1-2Manα1-3)Man, Manα1-6(Manα1-2Manα1-3)Man, Manα1-2Manα1-6(Manα1-3)Man, and Manα1-6(Manα1-3)Man in one-pot. Each oligomannosyl branch could be easily purified by liquid chromatography. The resulting structural isomers were identified by 2D-HMBC NMR. A systematic lectin affinity assay using the prepared oligomannosyl branches showed different specificities for the Galanthus nivalis lectin between structural isomers of the oligomannosyl branches with the same number of mannose residues..

Synthetic trisaccharides reveal discrimination of endo-glycosidic linkages by exo-acting α-1,2-mannosidases in the endoplasmic reticulum.

A tri-antennary Man9GlcNAc2 glycan on the surface of endoplasmic reticulum (ER) glycoproteins functions as a glycoprotein secretion or degradation signal after regioselective cleavage of the terminal α-1,2-mannose residue of each branch. Four α-1,2-mannosidases-ER mannosidase I, ER degradation-enhancing α-mannosidase-like protein 1 (EDEM1), EDEM2, and EDEM3-are involved in the production of these signal glycans. Although selective production of signal glycans is important in determining the fate of glycoproteins, the branch-discrimination abilities of the α-1,2-mannosidases are not well understood. A structural feature of the Man9GlcNAc2 glycan is that all terminal glycosidic linkages of the three branches are of the α-1,2 type, while the adjacent inner glycosidic linkages are different. In this study, we examined whether the α-1,2-mannosidases showed branch specificity by discriminating between different inner glycosides. Four trisaccharides with different glycosidic linkages [Manα1-2Manα1-2Man (natural A-branch), Manα1-2Manα1-3Man (natural B-branch), Manα1-2Manα1-6Man (natural C-branch), and Manα1-2Manα1-4Man (unnatural D-branch)] were synthesized and used to evaluate the hypothesis. When synthesizing these oligosaccharides, highly stereoselective glycosylation was achieved with a high yield in each case by adding a weak base or tuning the polarity of the mixed solvent. Enzymatic hydrolysis of the synthetic trisaccharides by a mouse liver ER fraction containing the target enzymes showed that the ER α-1,2-mannosidases had clear specificity for the trisaccharides in the order of A-branch > B-branch > C-branch ≈ D-branch. Various competitive experiments have revealed for the first time that α-1,2-mannosidase with inner glycoside specificity is present in the ER. Our findings suggest that exo-acting ER α-1,2-mannosidases can discriminate between endo-glycosidic linkages.

Design and synthesis of an amphiphilic graft hydrogel having a hydrophobic domain formed by multiple interactions.

A novel hydrogel having hydrophobic oligo segments and hydrophilic poly(acrylamidoglycolic acid) (PAGA) as pH responsive polymer segments was designed and synthesized to be used as a soft biomaterial. Poly(trimethylene carbonate) (PTMC) as the side chain, for which the degrees of polymerization were 9, 19, and 49, and the composition ratios were 1, 5, and 10mol%, was used as the oligo segment in the hydrogel. The swelling ratio of the hydrogel was investigated under various changes in conditions such as pH, temperature, and hydrogen bonding upon urea addition. Under pH2-11 conditions, the graft gel reversibly swelled and shrank due to the effect of PAGA main chain. The interior morphology and skin layer of the hydrogel was observed by a scanning electron microscope. The hydrogel composed of PAGA as the hydrophilic polymer backbone had a sponge-like structure, with a pore size of approximately 100µm. On the other hand, upon increasing the ratio of trimethylene carbonate (TMC) units in the hydrogel, the pores became smaller or disappeared. Moreover, thickness of the skin layer significantly increased with the swelling ratio depended on the incorporation ratios of the PTMC macromonomer. Molecular incorporation in the hydrogel was evaluated using a dye as a model drug molecule. These features would play an important role in drug loading. Increasing the ratio of TMC units favored the adsorption of the dye and activation of the incorporation behavior.

Gel formation driven by tunable hydrophobic domain: design of acrylamide macromonomer with oligo hydrophobic segment.

Nowadays, biomaterials with amphiphilic properties are undergoing remarkable development. Here, we present one such development, in which we prepared amphiphilic graft copolymers, with a main chain composed of hydroxyethyl acrylamide (HEAA), to introduce hydrophilicity, and a side chain composed of poly(trimethylene carbonate) (PTMC) to introduce tunable hydrophobicity. These macromonomers were created with a novel molecular design, which introduced a ring-opening polymerization by the hydroxyl end group of HEAA in the presence of 1,8-diazabicyclo[5.4.0]undec-7-ene, and were analyzed by (1)H NMR and gel permeation chromatography. The amphiphilic graft copolymers were shown to form a hydrogel, the swelling ratio of which was greatly influenced by the number of trimethylene carbonate units. These copolymers also exhibited the Tyndall phenomenon in aqueous solution; they aggregated spontaneously due to hydrogen bonding and hydrophobic interactions, and a sodium 8-anilino-1-naphthalenesulfonate (ANS) fluorescence probe was introduced into the hydrophobic domain. The solution property of ANS in the polymer solution was analyzed by fluorescence measurement and (1)H NMR. The maximum fluorescence wavelength of ANS shifted to shorter wavelengths as the degree of polymerization of the hydrophobic PTMC, the composition of the macromonomer, and the concentration of the copolymer increased. The resulting copolymer formed a polymer micelle structure due to the tunable hydrophobic domain formation in selected solvents. Therefore, these amphiphilic graft copolymers containing a PTMC segment are excellent candidates for use as hydrophobic drug delivery carriers.