13C and 1H solid state NMR investigation of hydration effects on gluten dynamics

The effect of hydration on the molecular dynamics of soft wheat gluten was investigated by solid state NMR. For this purpose, we recorded static and MAS ¹H spectra and SPE, CP, and other selective ¹³C spectra under MAS and dipolar decoupling conditions on samples of dry and H₂O and D₂O hydrated gluten. Measurements of carbon-proton CP times and several relaxation times (proton T₁, T_1ρ and T₂, and carbon T₁) were also performed. The combination of these techniques allowed both site-specific and domain-averaged motional information to be obtained in different characteristic frequency ranges. Domains with different structural and dynamic behaviour were identified and the changes induced by hydration on the dynamics of different domains could be monitored. The proton spin diffusion process was exploited to get information on the degree of mixing among different gluten domains. The results are consistent with the “loop and train” model proposed for hydrated gluten.     

Determination of the degree of acetylation of chitin materials by 13C CP/MAS NMR spectroscopy

¹³C CP/MAS NMR spectroscopy has been shown to be a powerful tool to quantify the degree of acetylation of chitin and chitosan. In order to optimise the parameters which afford quantitative ¹³C cross-polarisation magic-angle spinning NMR spectra, a detailed relaxation study has been carried out on selected chitin and deacetylated chitin samples. A relaxation delay of 5 s and a contact time of 1 ms have been found to yield quantitative NMR spectra of samples with deacetylation degree values of 0.68 and 0.16. The measured spin-lattice relaxation times in the rotating frame, T_1ρH, are in the range 6.4–8.9 ms for chitin and 4.3–7.3 ms for deacetylated chitin, while T_CH values for both samples are very similar and range from 0.03 to 0.19 ms. Spin-counting experiments indicate that, within experimental error, all carbon is detected by NMR indicating that the samples studied contain no (or very few) paramagnetic centres.     

Behavior of cellulose production of Acetobacter xylinum in C-enriched cultivation media including movements on nematic ordered cellulose templates

Acetobacter xylinum NQ-5 (ATCC 53582) and AY-201 (ATCC 23769) cultivated in the Hestrin–Schramm medium containing d-glucose with a natural ¹³C-abundance of 1.1% were investigated regarding their cell division rates and the bacterial movements. Comparative studies were carried out in the presence of d-glucose-U-¹³C₆ with a uniform ¹³C-labeling of 99% as the sole carbon source. The bacterial growth rates in numbers were found to increase in the ¹³C-enriched media by about 13% for NQ-5 and 26% for AY-201, respectively. The movements of single cells caused by the inverse force of the secretion and deposition of cellulose nanofibers on nematic ordered cellulose (NOC) templates were investigated by real-time video analyzes using light microscopy. As a result, d-glucose-U-¹³C₆ reduced the speed of motion for both strains, which was an opposite trend to the above growth rates in the cell division. The deposited cellulose fibers were proved to be in a cellulose crystalline form by CP/MAS ¹³C NMR and NIR FT Raman spectroscopic measurements, although the biosynthesized and thereafter-deposited cellulose has a strong interaction with the surface of NOC.     

Celery (Apium graveolens) parenchyma cell walls: cell walls with minimal xyloglucan

The primary walls of celery ( Apium graveolens L.) parenchyma cells were isolated and their polysaccharide components characterized by glycosyl linkage analysis, cross-polarization magic-angle spinning solid-state ¹³C nuclear magnetic resonance (CP/MAS ¹³C NMR) and X-ray diffraction. Glycosyl linkage analysis showed that the cell walls consisted of mainly cellulose (43 mol%) and pectic polysaccharides (51 mol%), comprising rhamnogalacturonan (28 mol%), arabinan (12 mol%) and galactan (11 mol%). The amounts of xyloglucan (2 mol%) and xylan (2 mol%) detected in the cell walls were strikingly low. The small amount of xyloglucan present means that it cannot coat the cellulose microfibrils. Solid-state ¹³C NMR signals were consistent with the constituents identified by glycosyl linkage analysis and allowed the walls to be divided into three domains, based on the rigidity of the polymers. Cellulose (rigid) and rhamnogalacturonan (semi-mobile) polymers responded to the CP/MAS ¹³C NMR pulse sequence and were distinguished by differences in proton spin relaxation time constants. The arabinans, the most mobile polymers, responded to single-pulse excitation (SPE), but not CP/MAS ¹³C NMR. From solid-state ¹³C NMR of the cell walls the diameter of the crystalline cellulose microfibrils was determined to be approximately 3 nm while X-ray diffraction of the cell walls gave a value for the diameter of approximately 2 nm.     

A study of ordered structure in acid-modified tapioca starch by C CP/MAS solid-state NMR

Acid modification of tapioca starch earlier reported to increase the mechanical strength of tablets. The development of ordered structure (double helices) of these starches was monitored after equilibrating at 0.90 a_w (25 °C) using ¹³C CP/MAS NMR and X-ray diffraction. As the hydrolysis time increased, the intensity of the resonance for C1 and C4 amorphous fractions decreased while that for C1 and C4 double helix fractions increased. Relative crystallinity (%) obtained from ¹³C CP/MAS NMR and X-ray diffraction methods both increased sharply initially and then levelled off with hydrolysis time. The initial increase in relative double helix content and crystallinity was due to a hydrolytic destruction in the amorphous domain, retrogradation of the partially hydrolyzed amylose and crystallization of free amylopectin double helices. After 192 h, these two parameters were not significantly different (=0.05) indicating that the double helices that were not arranged into crystalline regions either had been hydrolyzed or crystallized.     

Bradykinin effects on phospholipid metabolism and its relation to arachidonic acid turnovers in neuroblastoma × glioma hybrid cells (NG 108-15)

In neuroblastoma × glioma hybrid cells (NG 108-15) labelled with [³²P]-trisodium phosphate, [³H]-inositol and [¹⁴C]-arachidonic acid, bradykinin stimulated the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP₂) while it had no effect on the release of [¹⁴C]-arachidonic acid (AA). The effect on PIP, was time- and dose-dependent with a maximal effect on [³H]-inositol- and [³²P]-labelled cells after 10–30 s of stimulation with 10⁻⁶ M bradykinin. However, the hydrolysis of [¹⁴C]-AA labelled PIP₂ was delayed compared to the effect on [³H]- and [¹⁴C]-PIP₂ and was not detectable until after 60 s of stimulation. Bradykinin stimulation resulted in an increased formation of [³H]-inositol phosphates (IP) and [³²P]- and [¹⁴P]- and [¹⁴C]-phosphatidic acid (PA) but the time course for PA formation did not allow the time-course for PIP₂ hydrolysis. A reduced labelling of [²³P]- and [¹⁴C]-phosphatidylcholine was also found in stimulated cells suggesting that PA may derive from other sources than PIP₂. In conclusion, our results indicate that bradykinin activates phospholipase C, but not phospholipase A₂, in NG 108-15 cells.     

红壤和风沙土添加不同化学结构有机碳对外源碳去向及累积贡献的影响

植物凋落物碳输入显著影响陆地生态系统土壤CO₂排放和有机碳(SOC)形成,然而,针对不同质地土壤添加不同化学结构外源碳去向依然不清楚。本研究将¹³C标记的葡萄糖、淀粉和纤维素添加至红壤和风沙土,比较2种质地土壤添加不同化学结构外源碳在土壤释放的CO₂、SOC、可溶性有机碳(DOC)和微生物生物量碳(MBC)库的净累积量、回收率及贡献比例上的差异。结果表明: 添加外源有机碳显著提高了CO₂、SOC、DOC和MBC的δ¹³C值,且随着外源有机碳化学结构复杂性的增加,CO₂的δ¹³C峰值依次延迟出现;外源有机碳种类、土壤类型和培养时间均显著改变外源碳去向及其在各碳库的贡献比例;在风沙土中,外源有机碳更多被矿化为CO₂,且CO₂库的外源碳净累积量和回收率大小依次为葡萄糖>淀粉>纤维素;红壤添加外源碳转变为SOC的累积量和回收率显著高于风沙土,且红壤SOC库的外源碳净累积量和回收率大小顺序也为葡萄糖>淀粉>纤维素。可见,外源有机碳化学结构和土壤质地共同调控外源碳去向及累积贡献。     

Synthesis and characterization of a dianionic carbohydrate-based phospholipid

A novel carbohydrate-based phospholipid containing a phosphatidic acid head group, bis-(2,3-lauroyl)-1-methoxy-5-ribo-phosphatidic acid (DLRPA), was synthesized and characterized by ¹H NMR, ¹³C NMR, and ³¹P NMR. This molecule is an analog of dilauroyl phosphatidic acid (DLPA). The T_m of DLRPA decreases with increasing pH in a similar pattern to DLPA, as determined by MDSC. From this thermotropic behavior, the apparent pK_as of DLRPA are estimated to be 4 and 8.     

新疆阿勒泰地区近440年来大气δ13C变化

化石燃料的大量使用和森林的过度砍伐,引起大气中CO₂浓度的大幅度增加,同时由于Suess效应,大气CO₂中的δ¹³C在不断地下降。植物中δ¹³C的变化是大气CO₂浓度和同位素比值变化的敏感指示器。文中利用树木年轮δ¹³C序列和植物碳同位素分馏模型,尝试恢复了新疆阿勒泰地区近440年来大气δ¹³C的变化。结果表明,1850年之前,从树木年轮δ¹³C序列恢复的大气δ¹³C相对恒定在-6.60‰(R²=0.052),而1850年之后,该大气δ¹³C明显降低(R²=0.65),平均约为-7.04‰,平均年降低0.0084‰。这一结果高于从冰芯气泡所恢复的大气δ¹³C,1850年～1981年冰芯大气δ¹³C平均年降低约0.00657‰这可能与从树木年轮δ¹³C序列恢复的大气δ¹³C有更高的分辨率及树木生长点大气δ¹³C不同于全球大气δ¹³C值有关。     

Synthesis and characterization of trisodium salt of bis(thiosalicylato)aurate(I): Na3[Au(TSA)2]·5H2O

Sodium salt of a water-soluble, anionic, and monomeric 1:2 complex of Au(I) with a dianion of thiosalicylic acid TSA²⁻(Hin2TSA) = o-HS(C₆H₄)COOH) was first prepared and isolated as colorless needle crystals through a stoichiometric reaction of NaAuCl₄:H₂TSA:NaOH = 1:4:8 molar ratio in aqueous/EtOH solution. In this reaction, TSA²⁻ ligand has played a role of a reducing agent for the starting Au(III) ion and also of donor ligands coordinating to the reduced Au(I). This compound was characterized by complete elemental analyses, TG/DTA, FT-IR, 2D-NMR (¹H-¹H COSY, ¹H-¹³C HMBC, and ¹H-¹³C HMQC) spectroscopy, and the molmass measurement based on the cryoscopic method. It was shown that this complex was a monomeric species of Au(I) with a formula of Na₃[Au(TSA)₂]·5H₂O in the solid state, but not a polymeric species even in aqueous solution. A full assignment of seven carbon and four proton resonances in the coordinated TSA²⁻ ligand was achieved by the 2D ¹H-¹³C HMBC NMR technique.     

Mobility of lipid in complexes of amylose–fatty acids by deuterium and C solid state NMR

Palmitic and lauric acid complexes with amylose were studied by solid state methods: ¹³C CP/MAS NMR, deuterium NMR, X-ray powder diffraction and differential scanning calorimetry (DSC). The crystalline amylose complexes were found to be in a V-type sixfold single chain helix. The melting points of the complexes were over 100°C, at least 40–50°C higher than the melting points of the free fatty acids. CP/MAS ¹³C NMR spectra revealed two resonance peaks at 33.6 and 32.4 ppm for the palmitic acid, which were assigned as free and complexed fatty acid, respectively. A single resonance peak at 32.4 ppm was found for the lauric acid and assigned to the complex. The chemical shift of 32.4 ppm for the complexed fatty acids suggests a combined trans and gauche conformation for the fatty acid chain in the complex. T₁ relaxation measurements on the two palmitic acid resonances show different behavior: a very slow relaxation for the 33.6 ppm and a much faster relaxation (1.2 s) for the 32.4 ppm resonances. The latter was similar to the relaxation of the single resonance of the lauric acid (1.1 s). Temperature dependent deuterium spectra of the amylose–lauric acid and amylose–palmitic acid complexes suggest a complete complexation for the amylose–lauric acid, whereas the amylose–palmitic acid complex is partially disassociated by the thermal treatment. Based on the overall data, a partially disordered model is proposed: an imperfect helix with the fatty acid partly inside and partly out, depending on crystallization conditions and the necessity of placing the carboxyl head outside the V-helix.     

Autoradiographic mapping of pulmonary NK1 and NK2 tachykinin receptors and changes after repeated antigen challenge in guinea pigs

An autoradiographic technique was used to study the distribution of changes in pulmonary NK₁ and NK₂ receptors in guinea pig lung after repeated antigen challenge. Specific labeling of [³H] CP96345 (NK₁ receptors) and [³H] SR48968 (NK₂ receptors) was localized over the tracheal and bronchial smooth muscle; the density of binding increased towards smaller airways with a higher density for [³H] CP96345 binding. Bronchial epithelium and pulmonary blood vessels were also labeled densely with [³H] CP96345. No remarkable difference in the pattern of distribution of pulmonary NK₁ and NK₂ tachykinin receptors was observed between control, vehicle-challenged, and repeatedly antigen-challenged (weekly for three times) guinea pigs. A significant reduction in specific labeling of [³H] CP96345 (p < 0.01) and [³H] SR48968 (p < 0.05) over pulmonary structures was observed in antigen-challenged compared to control or vehicle-challenged animals. This study provides evidence that NK₁ and NK₂ tachykinin receptors are both localized to smooth muscle of all sizes in guinea pig airways and provides further evidence for a discrete distribution of NK₁ and NK₂ tachykinin receptors, consistent with their relative functional activities. In a established model of airway inflammation a decrease in the expression of NK₁ and NK₂ tachykinin receptors was evident on several different cell types within the lung, and this could influence airway and vascular reactivity.     

Coordination of a disubstituted alkene in a chelated d-metal-alkyl-alkene complex. Evidence for equilibrium between complexes with coordinated and free alkenes

Addition of (Cp*₂YH)₂ (4) to 2-methyl-1,4-pentadiene produced the yttrium-alkyl-alkene chelate complex Cp*₂YCH₂CH₂CH₂C(CH₃)=CH₂ (2) in which a disubstituted alkene is complexed to the metal center. Evidence for coordination of the alkene unit of 2 comes from the ¹H and ¹³C NMR chemical shifts of the vinyl units and from observation of nOe effects between Cp* protons and vinyl hydrogens. The disubstituted alkene ligand of 2 is weakly bound, and evidence for an equilibrium with substantial amounts of complex 3 with a free alkene was obtained from variable temperature ¹H NMR spectroscopy.     

A Regioselective, Chemoenzymatic Synthesis of Sucrose- 1'-Methacrylate

Pure sucrose is inexpensive and readily available, making this disaccharide a highly desirable starting material for new polymers. In order to achieve a clean polymerization, the disaccharide must be regioselectively monofunctionalized in good yield. This paper describes the practical, enzyme-mediated synthesis of sucrose-1 '-methacrylate 2 from sucrose and vinyl methacrylate using subtilisin Carlsberg (Sigma, Protease VIII), a readily available bacterial serine protease. A key aspect of this process, ascertaining the positional selectivity of acylation, was unambiguously accomplished using ¹H-detected (¹H, ¹³C) one-bond shift correlation (HMQC) spectroscopy and¹H-detected (¹H, ¹³C) multiple bond correlation (HMBC) spectroscopy.     

Biosynthesis of C-labeled branched polysaccharides by pestalotiopsis from C-labeled glucoses and the mechanism of formation

Biosynthesis of branched glucan by Pestalotiopsis from media containing D-(1-¹³C)glucose, D-(2-¹³C)glucose, D-(4-¹³C)glucose, D-(6-¹³C)glucose or a mixture of D-(1-¹³C)glucose and D-(2-¹³C)glucose was carried out to elucidate biosynthetic mechanism of branched polysaccharides. ¹³C NMR spectra of the labeled polysaccharides were determined and assigned. Analysis of ¹³C NMR spectra of glucitol acetates obtained from hydrolysates of the labeled branched polysaccharides indicated that transfer of labeling from C-1 to C-3 and C-6 carbons, from C-2 to C-1, C-3 and C-5 carbons, and from C-6 to C-1 carbon. From the results the percentages of routes via which the polysaccharide is biosynthesized are estimated. They show that the biosynthesis of the polysaccharide via the Embden-Meyerhof pathway and that from lipids and proteins are more active, and the pentose cycle is less active, than in the biosynthesis of cellulose and curdlan. As for the results, labeling at C-6 carbon in the branched polysaccharide cultured from D-(6-¹³C)glucose was low, compared to that of cellulose and curdlan.     

Synthesis, solution behaviour and molecular structures of 16-electron closo-2-(Ph3P)-1-N,2-[μ-(η-CH2CH=Ch2)]-1-N-(σ-CH2CH=CH2)-2,1- RhCB10H10, the first η-olefinic monocarbon metallacarborane

The first η²-olefinic monocarbon metallacarbone closo-2-(Ph₃P)-1-N,2-[μ-(η²-CH₂CH=Ch₂)]-1-N-(σ-CH₂CH=CH₂)-2,1- RhCB₁₀H₁₀ has been prepared by the reaction of the dimeric anion {[Ph₃PRhB₁₀H₁₀CNH₂]₂-μ-H}⁻[PPN]⁺ with allyl bromide and characterized by a combination of spectroscopic methods and a single-crystal X-ray diffraction study. The variable temperature ¹H and ¹³C NMR studies revealed the fluxional behavior of the η²-olefinic complex in CD₂Cl₂ solution which is associated with the allyl side-chain exchange process.     

测墒补灌条件下施氮量对小麦开花后13C同化物积累量和水氮利用效率的影响

为探究黄淮冬麦区测墒补灌节水条件下协同提高小麦产量和水氮利用效率的氮肥管理措施,以小麦品种‘烟农1212'为材料,在拔节期和开花期将各处理0～40 cm土层土壤相对含水量均补灌至70%条件下,设置3个施氮水平,即150(N₁)、210(N₂)和270 kg·hm^-2(黄淮冬麦区常规施氮量,N₃),研究施氮量对小麦开花后旗叶光合特性、¹³C同化物积累与转运及水氮利用效率的影响。结果表明: N₂和N₃处理开花后14～35 d旗叶光合能力显著高于N₁处理,N₂与N₃处理间差异不显著。¹³C同位素示踪结果显示,N₂处理开花后营养器官¹³C同化物转运量比N₁和N₃处理分别高12.1%和7.1%,成熟期¹³C同化物在籽粒中的分配量比N₁和N₃处理分别高10.1%和5.3%。施氮量亦调节了小麦不同生育阶段的耗水量、耗水模系数和总耗水量,小麦全生育期耗水量表现为N₂与N₃处理无显著差异,但均显著高于N₁处理,N₂处理拔节至成熟期阶段耗水量和耗水模系数均较高。N₂处理水分利用效率比N₃和N₁处理分别高7.5%和4.8%,籽粒产量比N₃和N₁处理分别高4.7%和10.9%,氮肥偏生产力比N₃处理高34.6%。综合考虑小麦籽粒产量和水氮利用效率,施氮量为210 kg·hm^-2处理为研究区测墒补灌节水条件下的最佳施氮量。     

High-resolution magic-angle spinning H nuclear magnetic resonance studies of lipid dispersions using spherical glass ampoules

A very useful high-resolution magic-angle spinning (MAS) ¹H NMR method for studying lipid dispersions is presented. The sample can be loaded into the spherical glass ampoule very easily, and a spinning speed of more than 10 kHz can be achieved without the problems of sample leakage or water loss. The line width at half height for the HDO peak is less than 1.5 Hz, and the method can be implemented by anyone who has access to a solid-state MAS NMR spectrometer. By using the spherical glass ampoule method we found two water peaks, which could be ascribed to bulk water outside of the multilamellar liposome (peak at high frequency) and interlamellar water (peak at low frequency), in both POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) liposomes. These are the first two examples of lipids without exchangeable protons that exhibit two distinct water resonances. The respective ¹H spin-lattice relaxation times (T₁) were measured, yielding values twice as long for bulk water as compared with interlamellar water. Both the chemical shift and spin relaxation results demonstrate the ability of MAS ¹H NMR to rapidly monitor changes in physical properties that accompany water interactions with zwitterionic phosphatidylcholines.     

Macrocylic thioether-esters and thioether-thioesters and their palladium, platinum and silver complexes

Preparations by the high dilution method are reported for seven macrocyclic thioether-esters and thioether-thioesters (L1–;L7). Yields in these reactions between thiodiglycolyl dichloride and appropriate ,ω-diols or dithiols range from 10 to 51%. The compounds are characterized by ¹H and ¹³C NMR, IR and high resolution mass spectroscopy. They react with salts of Pd(II), Pt(II) and Ag(I) to form complexes of which MX₂·L2, (M = Pt, X = Cl; M = Pd, X = Cl, Br, I, SCN), [Pd(L2)₂][CF₃SO₃]₂·H₂O and [Ag(L5)₂][CF₃SO₃]·C₂H₅OH have been isolated and characterized by elemental analysis, IR and NMR spectroscopy. NMR spectra indicate reversible dissociation of the ligand occurs in dimethyl sulfoxide solvent for PdCl₂·L2 but not for the Pt analogue. For PtCl₂·L2, spectra indicate that the ligand is undergoing a conformational ‘wag’ about its pair of equivalent sulfurs. These remain bound to the metal while the unique sulfur moves from the apical position of the coordination sphere to a non-coordinated situation. Simultaneously, inversions at the bound sulfurs are occurring.     

Application of CP-MAS and liquid-like solid-state NMR experiments for the study of the ripening-associated cell wall changes in tomato

¹³C and ¹H NMR spectra of an ethanol insoluble material (EIM) prepared from the pericarp of mature-green (MG) and red-ripe (RR) tomato fruits were acquired in ‘liquid-like’ and cross-polarisation with dipolar decoupling and magic angle spinning (CPMAS) conditions using the same triple resonance probe. Such a strategy allowed acquisitions of various NMR experiments aimed at detecting compositional differences as well as distinguishing differences in molecular mobility for various constituent polysaccharides related with the two ripening stages. Increase of the proton dipolar decoupling power levels from 3 to 50–55 kHz during single pulse ¹³C acquisition, led to more intense signals for pectic and hemicellulosic polysaccharides. This behaviour was interpreted as reflecting motional restrictions of these polysaccharides inside the porous cell wall network. Measurements of the proton rotating frame relaxation times T_1ρ in the ‘liquid-like’ conditions and of the proton transverse relaxation times T₂ from CPMAS spectra, revealed changes in mobilities for some pectic polysaccharides in relation with ripening, particularly for the H1 and H5 protons of -1,5 arabinan (Ara) side chains of rhamnogalacturonans. These data are discussed in relation with known pectic modifications occurring during ripening and associated with the tomato fruit softening.