Methylenedioxy- and methoxyflavones from Melicope coodeana syn. Euodia simplex

Three new natural products, 3,8-dimethoxy-5,7-dihydroxy-3′,4′-methylenedioxyflavone, 3,6,8-trimethoxy-5,7-dihydroxy-3′,4′-methylenedioxyflavone and 3,6,8,3′,4′-pentamethoxy-5,7-dihydroxyflavone were isolated from Melicope coodeana syn. Euodia simplex (Rutaceae) along with 3,6,3′-trimethoxy-5,7,4′-trihydroxyflavone and 3,3′-dimethoxy-5,7,4′-trihydroxyflavone. The structural assignments are based on ¹H and ¹³C NMR data, including discussion of the chemical shifts of C-2 in 3,5-dihydroxy- and 3-methoxy-5-hydroxyflavones. The presence of highly methoxylated and methylenedioxyflavones is characteristic of the genus Melicope, and the present findings support the recent transfer of Euodia simplex to Melicope.     

Sn and C NMR study of some trivinyltin(IV) compounds and their complexes

The¹¹⁹Sn and ¹³C NMR spectra of ten trivinyltin(IV) compounds in solutions of non-coordinating (deuteriochloroform, trideuterionitromethane) and coordinating (hexadeuteriodimethyl sulphoxide) solvents have been studied. From δ(¹¹⁹Sn) chemical shifts and ¹J(¹¹⁹Sn,¹³C) coupling constants an evaluation of the coordination number of the central tin atom and the shape of coordination polyhedra around the tin atom has been carried out. Various effects on the δ(¹³C) chemical shifts of both carbon atoms of the vinyl group are also discussed.     

Changes in drug 13C NMR chemical shifts as a tool for monitoring interactions with DNA

The antibiotic drug, netropsin, was complexed with the DNA oligonucleotide duplex [d(GGTATACC)]2 to monitor drug 13C NMR chemical shifts changes. The binding mode of netropsin to the minor groove of DNA is well-known, and served as a good model for evaluating the relative sensitivity of 13C chemical shifts to hydrogen bonding. Large downfield shifts were observed for four resonances of carbons that neighbor sites which are known to form hydrogen bond interactions with the DNA minor groove. Many of the remaining resonances of netropsin exhibit shielding or relatively smaller deshielding changes. Based on the model system presented here, large deshielding NMR shift changes of a ligand upon macromolecule binding can likely be attributed to hydrogen bond formation at nearby sites.     

Incorporation of Petroleum Carbon Into Soil Organic Matter During Biodegradation

A procedure, based on measurement of the stable carbon isotope ¹³C, has been developed for determining the extent to which petroleum carbon is incorporated into soil organic matter (SOM) by humification of biomass produced during biodegradation of the petroleum in soil. We have shown that a crude oil having a δ¹³C of-27.4%, when biodegraded in a soil containing SOM with a δ¹³C of-15.7%, resulted in a change of the δ¹³C of the bound SOM reflecting that of petroleum carbon. Comparison of five soil biodegradation tests using different amounts and types of fertilizer to stimulate biodegradation of the oil in this soil showed that the extent of the δ¹³C change in the bound SOM varied with the extent of oil biodegradation observed. To obtain ¹³C data on the SOM, the residual petroleum was first removed by rigorous extraction with dichloromethane using a Soxhlet apparatus. The extracted soil was then combusted to release bound carbon as CO₂, which was analyzed for ¹³C. Where the SOM has a δ¹³C similar to that of petroleum, ¹⁴C measurements of SOM would give similar results. This type of data, referred to as the petroleum “footprint” in the SOM, could be useful in identifying or confirming intrinsic biodegradation of petroleum in contaminated soil.     

Activation of [C]chlorobiphenyls to protein-binding metabolites by rat liver microsomes

The irreversible binding of [¹⁴C]2,2′-di- and [¹⁴C]2,4,5,2′,4′,5′-hexachlorobiphenyl ([¹⁴C]DCB and [¹⁴C]HCB) to protein was studied in the presence of rat liver microsomes and a NADPH-generating system. Protein-bound radioactivity was found with [¹⁴C]DCB but not with [¹⁴C]HCB. The binding of ¹⁴C-metabolites was increased by pretreatment of the rats with phenobarbital or polychlorinated biphenyls. Protein binding was linear for 80 min. In contrast, monohydroxy-metabolites of DCB were formed and degraded within 40 min. Inhibition of secondary oxidation of DCB by scavening superoxide anions or by glucuronidation of the monophenols markedly decreased the protein binding. Addition of trichloropropene oxide or styrene oxide, both inhibitors of epoxide hydrase, did not significantly stimulate the binding. The results suggest that the majority of reactive metabolites of DCB arise from secondary metabolism, i.e., the subsequent oxidation of the phenolic metabolites. Arene oxides, the primary products, appear to play a minor role in the protein binding of DCB.     

Sources and accumulation rates of organic carbon in an equatorial peat bog (Burundi, East Africa) during the Holocene: carbon isotope constraints

The relationship between organic carbon accumulation rates and ¹³C/¹²C ratios of total organic carbon (TOC) was investigated in an highland peat bog core (Ru-3) from Equatorial Africa. This core yielded a sequence spanning the last 14 kyr and was analysed with a 100–300 yr resolution for TOC-δ¹³C values. The Holocene section shows contrasted TOC accumulation regimes and TOC δ¹³C varying between −28.5 and −19.5‰ with a few very short ‘isotopic excursions' (dated at ca. 9.3, 7.5, 4.2 ka B.P.). The organic carbon accumulation rates range from 2 to 20 mg C cm⁻² yr⁻¹. They increase when TOC becomes more depleted in ¹³C, notably between 12 and 9.8 ka B.P., 8.5 and 7.8 ka B.P. and after 1.6 ka B.P. Periods of restricted carbon storage correspond to heavier TOC accumulation at 9.3, and between 7.5 and 1.6 ka B.P. At the study site, the δ-variations can be related to variable C4-plant inputs, and possibly, to changes in the fractionation between CO₂ and the organic carbon in C3 vascular plants. The Ru-3 record indicates restricted carbon storage during the periods of increased contribution from C4 plants and/or of decreased fractionation between CO₂ and organic carbon in C3 plants. Changes in TOC-δ¹³C values in core Ru-3 seem to match fluctuations of East Equatorial African lakes. High lake stands correspond to low δ¹³C intervals and vice versa. This points to indirect climatic forcing of δ¹³C changes in intertropical peats.     

Variation in δC values for the seagrass Thalassia testudinum and its relations to mangrove carbon

Carbon isotope ratios (¹³C/¹²C) were measured for the leaves of the seagrass Thalassia testudinum Banks ex König and carbonates of shells collected at the seagrass beds from seven sites along the coast of southern Florida, U.S.A. The δ¹³C values of seagrass leaves ranged from −7.3 to −16.3‰ among different study sites, with a significantly lower mean value for seagrass leaves from those sites near mangrove forests (−12.8 ± 1.1‰) than those far from mangrove forests (−8.3 ± 0.9‰; P < 0.05). Furthermore, seagrass leaves from a shallow water area had significantly lower δ¹³C values than those found in a deep water area (P < 0.01). There was no significant variation in δ¹³C values between young and mature leaves (P = 0.59) or between the tip and base of a leaf blade (P = 0.46). Carbonates of shells also showed a significantly lower mean δ¹³C value in the mangrove areas (−2.3 ± 0.6‰) than in the non-mangrove areas (0.6 ± 0.3‰; P <0.025). In addition, the δ¹³C values of seagrass leaves were significantly correlated with those of shell carbonates (δ¹³C seagrass leaf = −9.1 + 1.3δ¹³C shell carbonate (R² = 0.83, P < 0.01)). These results indicated that the input of carbon dioxide from the mineralization of mangrove detritus caused the variation in carbon isotope ratios of seagrass leaves among different sites in this study.     

Nuclear magnetic resonance spectroscopy of 3 beta,7 beta-dihydroxy-5-cholen-24-oic acid multi-conjugates: unusual bile acid metabolites in human urine

Complete ¹H and ¹³C resonance assignments were made for a new type of 3β,7β-dihydroxy-5-cholen-24-oic acid doubly conjugated with sulfuric acid at C-3 and N-acetylglucosamine at C-7 and its glycine- and taurine-amidated triple-conjugates by the combined use of several homonuclear and heteronuclear shift-correlated 2D NMR techniques. The effects of sulfation at C-3, N-acetylglucosaminidation at C-7, and aminoacyl amidation at C-24 on the ¹H and ¹³C chemical shifts and signal multiplicity were clarified. The shielding data serving to characterize each of the bile acid multi-conjugates are also discussed.     

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.     

Phytochemical investigation on Mercurialis annua

A new natural flavonol glycoside, isorhamnetin-3-rutinoside-4′-glucoside, together with rutin, narcissin, quercetin-3-(2^G-glucosyl)-rutinoside and isorhamnetin-3-rutinoside-7-glucoside, was identified from the MeOH extract of Mercurialis annua L. The structures were established on the basis of chemical and spectral (¹H and ¹³C NMR, FAB MS) data.     

Effect of acyl donor chain length and substitutions pattern on the enzymatic acylation of flavonoids

Rutin and esculin were enzymatically acylated with different aliphatic acids as acyl donors (fatty acids, dicarboxylic acids and ω-substituted fatty acids) by an immobilized lipase from Candida antarctica. The effect of the water content and the acyl donors pattern on the flavonoid initial acylation rate and conversion yield were investigated. The obtained results indicated that the water content of the medium has a strong effect on the performance of these reactions. The best conversion yields were reached when the water content was kept lower than 200 ppm. At low water content of the medium, these syntheses are influenced by carbon chain length and substitution pattern of the acyl donors. Higher conversion yields of esculin and rutin (>70%) were obtained with aliphatic acids having high carbon chain length (>12). Moreover, it has been found that the amine and thiol groups on ω-substituted fatty acid chain were unfavourable to these reactions. The ¹H NMR and ¹³C NMR analyses of some synthesized esters (esculin and rutin palmitate) show that only monoesters were produced and that the esterification takes place on the primary OH of glucose moiety of the esculin and on the secondary 4′′′-OH of the rhamnose residue of rutin.     

Monitoring Anaerobic Natural Attenuation of Petroleum Using a Novel In Situ Respiration Method in Low-Permeability Sediment

Assessing petroleum biodegradation rates is an important part of predicting natural attenuation in subsurface sediments. Monitoring carbon dioxide (CO₂) and methane (CH₄) produced in situ, and their radiocarbon ¹⁴C), stable carbon (¹³C) and deuterium (D). signature provide a novel method to assess anaerobic microbial processes. Our objectives were to: (1) estimate the rate of anaerobic petroleum hydrocarbon (PH) mineralization by monitoring the production of soil gas CH₄ and CO₂ in the vadose zone of low-permeability sediment, (2) evaluate the dominant microbial processes using δ¹³C and δD, and (3) determine the proportion of CH₄ and CO₂ attributable to anaerobic mineralization of PH using ¹⁴C analysis. Argon was sparged into the subsurface to dilute existing CO₂ and CH₄ concentrations. Vadose zone CO₂, CH₄, oxygen, total combustible hydrocarbons, and argon concentrations were measured for 75 days. CO₂ and CH₄ samples were collected on day 86 and analyzed for ¹⁴C, δ¹³C, and δD. Based on CH₄ soil gas production, the anaerobic biodegradation rate was estimated between 0.017 to 0.055 mg/kg soil-d. CH₄ ¹⁴C (2.6 pMC), δ¹³C (-45.64‰), and δD (-316‰) values indicated that fermentation of PH was the sale source of CH₄ in the vadose zone. CO₂ ¹⁴C (62 pMC) indicated that approximately 47% of the total CO₂ was from PH mineralization and 53% from plant root respiration. Although low-permeability sediment increases the difficulty of completely replacing in situ soil gas and assuring anaerobic conditions, this novel respiration method distinguished between anaerobic processes responsible for PH degradation.     

Synthesis and NMR structural analysis of O-succinyl derivative of low-molecular-weight κ-carrageenan

Low-molecular-weight (LMW) κ-carrageenan was achieved through mild hydrochloric acid hydrolysis of κ-carrageenan. The acylation of LMW κ-carrageenan was performed by use of tetrabutylammonium (TBA) salt of the anionic polysaccharide fragments, succinic anhydride, 4-dimethylaminopyridine and tributylamine under homogeneous conditions in N,N-dimethylformamide at 80 °C. Investigation of FT-IR spectrum of the succinylated LMW κ-carrageenan showed that a monoester derivative with succinyl group was formed when LMW κ-carrageenan reacted with succinic anhydride. The ¹H and ¹³C NMR spectroscopy has been used to characterize the fine structure of O-succinyl derivative of the LMW κ-carrageenan. The ¹³C and ¹H NMR chemical shifts of disaccharide unit of O-succinyl LMW κ-carrageenan have been fully assigned using 2D NMR spectroscopic techniques.     

NMR studies of [U-C]cyclosporin A bound to human cyclophilin B

NMR data (¹H and ¹³C chemical shifts, NOEs) on [[U-¹³C]cyclosporin A bound to cyclophilin B were compared to previously published data on the [U-¹³C]CsA/CyPA complex [Fesik et al., (1991) Biochemistry 30, 6574–6583]. Despite only 64% sequence identity between CyPA and CyPB, the conformation and active site environment of CsA when bound to CyPA and CyPB are nearly identical as judged by the similarity of the NMR data.     

Pleistocene paleoenvironmental evolution at continental middle latitude inferred from carbon and oxygen stable isotope analysis of ostracodes from the Guadix-Baza Basin (Granada, SE Spain)

A representative paleoenvironmental reconstruction of continental middle latitude from ca. 2 my to the upper part of Middle Pleistocene (279 ± 77 ky) was obtained from the carbon and oxygen stable isotopes analyzed in ostracode shells (Cyprideis torosa) recovered in the Guadix-Baza Basin (SE Spain), an intramontaneous closed depression filled by alluvial and lacustrine sediments. This study was performed along a 356-m-thick composite section, dated previously by paleomagnetism and the amino acid racemization method. δ¹³C and δ¹⁸O profiles reflected changes in temperature, the evaporation/infill ratio in the water bodies and the amount of rain. δ¹³C is also affected by changes in plant biomass: periods with high δ¹³C and δ¹⁸O values are associated with warm and dry regimes, and with less vegetation, which, in some cases, coincide with the development of displacive gypsum crystals, whereas low δ¹³C and δ¹⁸O values correlate with cold and humid episodes, which cause more vegetation biomass and, therefore, increasing the input of isotopically light carbon. Intermediate δ¹⁸O values are linked to temperate dry or humid episodes when they coincide with high or low δ¹³C values, respectively. 86 paleoclimatic events were distinguished in the Pleistocene record from the δ¹³C and δ¹⁸O profiles. From both the statistical analysis of the geochemical data and the geological observations, four Cold and Humid Long Periods (low δ¹⁸O) and four Warm and Dry Long Periods (high δ¹⁸O) were defined. This differs with respect to the paleoclimatological behavior established for the Northern Hemisphere where during cold periods (glacial), no water was available while permafrost conditions persisted, whereas in warm episodes (interglacial), higher precipitation rates occurred. Good correspondences between the Guadix-Baza Basin paleoclimatic record and a marine oxygen-isotope sequence, two continental cores and other long Mediterranean paleoenvironmental records (pollen sequences from Israel) were found, which suggested that climate changes in the Guadix-Baza Basin were in tune with global climatic changes.     

Late Quaternary paleoproductivity changes off the Congo deduced from stable carbon isotopes of planktonic foraminifera

A detailed record (≈2-kyr intervals) of the difference in stable carbon isotopes (Δδ¹³C) between Globigerina bulloides and Globigerinoides ruber (pink) is used to reconstruct changes in upwelling intensity off the Congo River for the last 190,000 yr. Comparisons of the oxygen and carbon isotope data from this core with records from the Niger Fan and from pelagic cores in the eastern equatorial South Atlantic indicate that the Congo Fan isotope records do not contain a strong freshwater signal as is described off other major rivers. The temporal pattern of the Δδ¹³C signal correlates with the marine organic carbon record from the Congo Fan. Thus the planktonic Δδ¹³C record, reflecting past changes in upwelling intensity and nutrient content, corroborates the signal provided by sedimentary organic carbon, which is presumed to indicate changes in the amount of biological productivity and export flux to the seafloor.

The planktonic Δδ¹³C signal is characterized by a dominant 23-kyr periodicity which provides evidence for a strong response of upwelling fluctuations off the Congo to precessional forcing. Minima in the Δδ¹³C record are aligned with periods of minimum boreal summer insolation over Central Africa reflecting an increase of upwelling and biological productivity off the Congo at periods of enhanced zonal intensity of southeast trades and corresponding weak southeast monsoon over the eastern South Atlantic. A strong response to changes in river discharge probably did not occur, indicating that fertilization by river-derived nutrients has played only a minor role with respect to Late Quaternary changes in the total amount of primary productivity off the Congo.     

马尾松稳定碳同位素(δ13C)的地理变异及其对水热因子的响应

利用设置在浙江省淳安县姥山林场和湖北省太子山石龙林场2个试验点的33年生马尾松种源试验林,选取不同纬度的10个代表性种源,研究其在种源间的差异、地理变异模式及对水热因子的响应.结果表明: 马尾松平均年轮δ¹³C在种源间的差异极显著,高纬度种源的平均年轮δ¹³C高于低纬度种源.马尾松年轮δ¹³C呈纬向的地理变异模式,形成了对种源原地环境的适应性.年轮δ¹³C与种源地年均温(MAT)、1月均温(T₁)、年降水量(MAP)、5—9月降水量(P_5-9)及≥10 ℃年积温(CT)均呈显著或极显著负相关,与干燥度指数(AI)呈显著正相关.淳安和太子山点马尾松年轮δ¹³C对种源地干燥度指数的响应函数可分别解释年轮δ¹³C变化的37.5%和42.5%,种源地AI是年轮δ¹³C适应性的重要环境限制因子.马尾松稳定碳同位素δ¹³C的年际变化与生长环境关系密切,湖北太子山试验点处于相对干旱的中西部地区,干燥度指数高,平均年轮δ¹³C比淳安试验点高1.8%.太子山点和淳安点的马尾松年轮δ¹³C分别对7月和8月的气温响应敏感,夏季降水量是年轮稳定碳同位素分馏的主要限制因子,而不同种源对未来气候变化的响应敏感性不同.     

Sesquiterpene polyol esters from Celastrus rosthornianus

Two new sesquiterpene polyol esters with β-dihydroagarofuran skeleton were isolated from the root bark of Celastrus rosthornianus. Their structures were elucidated, mainly on the basis of spectral analyses, as 1 β-acetoxy-8β,9-dibenzoyloxy-6-hydroxy-2β(-methylbutanoyloxy)-β-dihydroagarofuran and 1β-acetoxy-9-benzoyloxy-8β-(β-furanocarbonyloxy)-6-hydroxy-2β(-methylbutanoyloxy)-β-dihydroagarofuran. The complete assignments of ¹³C NMR chemical shifts for both compounds on the basis of ¹H-¹³C chemical-shift correlation spectrum were also carried out.     

NMR study of paper

High quality antique sheets of paper have been characterized by ¹H NMR relaxations and ¹³C CP MAS spectra. Paper can be regarded as a bicomponent material made of cellulose and water plus a small amount of organic and inorganic impurities. Semicrystalline fibrous cellulose, rich in water, is present in the I and I_β forms. The amorphous cellulose, with a low water content, contains a higher amount of paramagnetic impurities and it is characterized by quite short ¹H spin-lattice relaxations and by ¹¹³C resonances with noticeable chemical shifts. Ad hoc tailored pulse sequences are able to produce ¹³C CP MAS spectra in which only the amorphous content of paper is clearly observed. It is shown that water is fully bound to the cellulose lattice. It also seems reasonable to formulate the hypothesis that a larger concentration of paramagnetic ions is located in the amorphous fraction of highly degraded paper compared with paper in good condition.     

高寒草甸不同坡向条件下植物叶片δ13C及水分利用效率的变化

通过对高寒草甸不同坡向条件下25科86种C3植物叶片稳定性碳同位素组成(δ¹³C)的测定,研究了高寒草甸C3植物δ¹³C和水分利用效率对坡向变化的响应以及环境因子对其产生的影响,分析了坡向上控制植物δ¹³C变化的主要环境因子.结果表明: 从北坡到南坡的生境梯度上,土壤含水量不断减少,而土壤温度及光照强度不断增加,植物群落结构也发生了相应变化.5个坡向中,植物叶片δ¹³C值为-31.19‰~-21.80‰,均值为(27.18±0.13)‰;生长季植物叶片δ¹³C均值在南坡最高,其次为西南坡、西坡、西北坡,最低值出现在北坡.坡向间δ¹³C值的差异主要是由不同坡向土壤含水量和土壤温度,以及光照强度的不同导致的,其中土壤含水量是主要的限制因子.北坡-南坡梯度上植物叶片δ¹³C值随土壤含水量下降、土壤温度及光照强度升高而变大,表明不同坡向植物的水分利用状况对干旱胁迫的响应不同,植物逐渐提高了水分利用效率以适应干旱胁迫的生境.