Turnover of “new” and “old” carbon in soil microbial biomass

The contributions of “new” carbon coming from plants with the C4-type of photosynthesis (maize) and “old” carbon from soil organic matter (SOM) formed under C3 vegetation as carbon sources for microorganisms was determined. Soil samples were taken from the plots of field experiments on Chernozem and Phaeozem. The values of δ¹³C were determined in evolved CO₂, SOM, total microbial biomass (C_mic), and phospholipid fatty acids (PLFA), assuming that the PLFA markers for certain taxonomic groups of microorganisms enriched in C4 carbon indicated a more significant role of these microorganisms in the transformation of root exudates and plant residues. Carbon pools were arranged in the following order by the degree of their enrichment with “new” C: SOM < C_mic < CO₂. Consequently, the “new” carbon proved to be a more preferable substrate for microbial growth than the “old” one. The share of C4 in the markers varied from 18 to 60% (on average 38%) in Phaeozem and from 15 to 40% in Chernozem (on average 28%). The groups of microorganisms in Phaeozem were arranged in the following order by the degree of their enrichment with “new” carbon: protozoa < saprotrophic fungi < actinomycetes < gram-positive bacteria < gramnegative bacteria < mycorrhizal fungi. In Chernozem, the contribution of C4 to the carbon composition of PLFA did not differ significantly for various groups of microorganisms. The C4 content within the PLFA markers of fungi and gram-negative bacteria did not demonstrate any crucial contribution of these groups of organisms to the transformation of “new” C. The long-term C3–C4 transition probably results in formation of a broad range of carbon pools similar in their C4 content but different in resistance to mineralization; therefore, gram-positive bacteria could assimilate C4 from resistant C pools. The low content of “new” carbon in the PLFA markers of fungi may be explained by a considerable portion of dormant forms.     

Schistosoma mansoni: circulating antigens and immune complexes in infected mice.

Circulating schistosome antigens (CSA) and circulating immune complexes (CIC) were investigated during the course of Schistosoma mansoni infection in mice. The radioimmunoprecipitation-polyethylene glycol (PEG) assay (RIPEGA) with [¹²⁵I]anti-S. mansoni antibodies or [¹²⁵I] anti-antigen “4” antibodies detected, respectively, total CSA and antigen “4” in serum and in 3% polyethylene glycol-precipitated CIC from infected mice. Complement fixation test and [¹²⁵I] C_1q-binding test revealed, respectively, an anticomplementary activity and the presence of C_1q-binding CIC. All these substances appeared in infected mice at approximately the same period, i.e., between the 40th- and the 55th- day postinfection. No correlation was observed between the detection of anticomplementary active substances and C_1q-binding CIC. In contrast, a close relationship was noticed between CSA and complement-activating material during the course of the infection. This suggests that substances with anticomplementary activity in serum from infected mice could be one or various CSA. A close correlation was also observed between C_1q-binding CIC and free or “complexed” antigen “4.” This observation supports well the possibility that antigen “4” is one of the major complexed circulating antigen present in schistosomiasis. The immunoglobulins G₁, G_2a, M, and A were also characterized in 3% PEG-precipitated CIC from infected mice during the period in which we detected C_1q-binding CIC. The roles played by specific S. mansoni CIC in either schistosomal nephropathy or protective mechanisms to a challenge infection in mice are discussed.     

Climatic information recorded in stable carbon isotopes in tree rings of Cryptomeria fortunei,Tianmu Mountain,China

The Cryptomeria fortunei (CF) tree-ring δ¹³C_p series, which was collected from the West Tianmu Mountain forestland (30°20′ N, 119°26′ E), located in the north-west of Zhejiang Province, China, belonging to the northern margin of the mid-subtropical region of Eastern China, were determined based on cross-dated tree-ring age. There was a significant decline in the δ¹³C_p series occurring from 1685 to 1985, more especially from 1835 to 1985 in response to increasing atmospheric CO₂ concentrations and decreasing atmospheric δ¹³C_a. To reduce the noise and enhance the climatic signals, we compared the polynomial function with the correction method developed by McCarroll and Loader (2004) to remove the low-frequency variation in the raw tree ring δ¹³C_p series (defined as the δ¹³C_poly series, δ¹³C_cor series, respectively), and found the most suited correction method was the correction method developed by McCarroll and Loader (2004) in our study area. High-frequency correlation analysis between the δ¹³C_cor series and many meteorological parameters recorded by Xian Rending weather station revealed that the current August–September mean maximum temperature and previous year mean minimum and mean maximum temperature (P < 0.005) most strongly influenced tree ring δ¹³C_p discrimination from 1956 to 1985, and the strongest temperature signal captured was the current August–September mean maximum temperature (r = 0.54, P < 0.005). Mainly on this basis, the varied history of current August–September mean maximum temperatures in the West Tianmu Mountain area were reconstructed from 1685 to 1985. The reconstructed maximum temperatures revealed a slight warming trend and showed close correlation with the climatic fluctuations of the Little Ice Age cold period before 1900 as well as the 20th century warm period after 1900. It also better corresponded with some climate events recorded in historical records. Spectrum analysis showed that in the reconstructed series there was quasi-periodicity of 66.7 yr, 21.1 yr, 3.2 yr, 2.3 yr and 2.0 yr. These cycles coincided with the “torque effect” variation of planets and the geocentric convergence, and changes in solar activity and irradiance, as well as the “Quasi-biennial oscillation” (QBO). This indicated that the δ¹³C_p chronology of tree rings in West Tianmu Mountain showed a good record of the sun's activities, the change in the sun radiation and ENSO events.     

Modification of the canonical profile transport model on the basis of new DIII-D experiments

Recent DIII-D experiments have shown that the stiffness of the ion temperature profile κ _i ^PC in the region 0.4 < ρ < 0.7 increases by one order of magnitude with increasing radius. At ρ < 0.4, the stiffness is low and the ion temperature profile is “soft.” The stiffness of the temperature profile also increases with decreasing the toroidal rotation velocity. The approximation of the experimental stiffness profiles allows one to modify the canonical profile transport model. The heat conductivity κ _i ⁰ in the plasma core is determined by minimizing the r.m.s. deviations of the calculated ion temperature from the measured one. This procedure also makes it possible to determine how κ _i ⁰ depends on the central ion temperature.     

Separation of targeted ganoderic acids from Ganoderma lucidum by reversed phase liquid chromatography with ultraviolet and mass spectrometry detections

Ganoderic acids are valuable bioactive secondary metabolites produced by a traditional medicinal mushroom Ganoderma lucidum (“Ling-zhi” in Chinese and “Reishi” in Japanese). In this work, a fast and efficient method for the recovery and purification of ganoderic acid T (GA-T) and ganoderic acid Me (GA-Me) from triterpene-enriched extracts of G. lucidum mycelia was developed by using reversed phase HPLC (RP-HPLC) on a C₁₈ column with an acidified methanol–water mobile phase in combination with ultraviolet (UV) detection and electrospray ionization mass spectrometry (ESI-MS). The presence of each targeted GA (GA-T and GA-Me) in its corresponding peak was easily identified and confirmed by UV and MS. The chemical structures of the purified GA-T and GA-Me were further confirmed by ¹H NMR. The retention behaviors of the two GAs over a temperature range of 15–55 °C were also investigated. From the retention time data, van’t Hoff plots were obtained. The estimated enthalpy (ΔH) and entropy (ΔS) data suggest that the retention time difference between GA-T and GA-Me might be driven by an enthalpy difference. Furthermore, a semi-preparative HPLC purification was achieved on a semi-preparative C₁₈ column using the conditions optimized for the analytical column. The method presented in this work can be a valuable tool for the rapid semi-preparative purification of targeted GAs, and it may also be applicable to some other natural products.     

Rhizobium indicum sp. nov., isolated from root nodules of pea (Pisum sativum) cultivated in the Indian trans-Himalayas

Three strains of rhizobia isolated from effective root nodules of pea (Pisum sativum L.) collected from the Indian trans-Himalayas were characterized using 16S rRNA, atpD and recA genes. Phylogeny of the 16S rRNA genes revealed that the newly isolated strains were members of the genus Rhizobium with ≥99.9% sequence similarity to the members within the “Rhizobium leguminosarum” group. Phylogenetic analyses based on the concatenated sequences of atpD and recA gene, and 92 core genes extracted from the genome sequences indicated that strains JKLM 12A2^T and JKLM 13E are grouped as a separate clade closely related to R. laguerreae FB206^T. In contrast, the strain JKLM 19E was placed with “R. hidalgonense” FH14^T. Whole-genome average nucleotide identity (ANI) values were 97.6% within strains JKLM 12A2^T and JKLM 13E, and less than 94% with closely related species. The digital DNA-DNA hybridization (dDDH) values were 81.45 within the two strains and less than 54.8% to closely related species. The major cellular fatty acids were C_18:1w7c in summed feature 8, C_{14:0 3OH}/C_{16:1 iso I} in summed feature 2, and C_18:0. The DNA G + C content of JKLM 12A2^T and JKLM 13E was 60.8 mol%. The data on genomic, chemotaxonomic, and phenotypic characteristics indicates that the strains JKLM 12A2^T and JKLM 13E represent a novel species, Rhizobium indicum sp. nov. The type strain is JKLM 12A2^T (= MCC 3961^T = KACC 21380^T = JCM 33658^T). However, the strain JKLM 19E represents a member of “R. hidalgonense” and the symbiovar viciae.     

Bacterial production and refolding from inclusion bodies of a “Weak” toxin, a disulfide rich protein

The gene for the “weak” toxin of Naja kaouthia venom was expressed in Escherichia coli. “Weak” toxin is a specific inhibitor of nicotine acetylcholine receptor, but mechanisms of interaction of similar neurotoxins with receptors are still unknown. Systems previously elaborated for neurotoxin II from venom of the cobra Naja oxiana were tested for bacterial production of “weak” toxin from N. kaouthia venom. Constructs were designed for cytoplasmic production of N. kaouthia “weak” toxin in the form of a fused polypeptide chain with thioredoxin and for secretion with the leader peptide STII. However, it became possible to obtain “weak” toxin in milligram amounts only within cytoplasmic inclusion bodies. Different approaches for refolding of the toxin were tested, and conditions for optimization of the yield of the target protein during refolding were investigated. The resulting protein was characterized by mass spectrometry and CD and NMR spectroscopy. Experiments on competitive inhibition of ¹²⁵I-labeled α-bungarotoxin binding to the Torpedo californica electric organ membranes containing the muscle-type nicotine acetylcholine receptor (α1₂β1_γδ) showed the presence of biological activity of the recombinant “weak” toxin close to the activity of the natural toxin (IC₅₀ = 4.3 ± 0.3 and 3.0 ± 0.5 µM, respectively). The interaction of the recombinant toxin with α7 type human neuronal acetylcholine receptor transfected in the GH₄C₁ cell line also showed the presence of activity close to that of the natural toxin (IC₅₀ 31 ± 5.0 and 14.8 ± 1.3 µM, respectively). The developed bacterial system for production of N. kaouthia venom “weak” toxin was used to obtain ¹⁵N-labeled analog of the neurotoxin.     

Salinicoccus halitifaciens sp. nov., a novel bacterium participating in halite formation

Strain JC90^T was isolated from a soda lake in Lonar, India. Strain JC90^T maintains its external pH to 8.5 and participates in halite formation. Based on 16S rRNA gene sequence similarity studies, strain JC90^T was found to belong to the genus Salinicoccus and is most closely related to “Salinicoccus kekensis” K164^T (99.3 %), Salinicoccus alkaliphilus T8^T (98.4 %) and other members of the genus Salinicoccus (<96.5 %). However Strain JC90^T is <36 % related (based on DNA–DNA hybridization) with the type strains of “S. kekensis” K164^T and S. alkaliphilus T8^T. The DNA G+C content of strain JC90^T was determined to be 46 mol %. The cell-wall amino acids were identified as lysine and glycine. Polar lipids were found to include diphosphatidylglycerol, phosphatidylglycerol, phosphatidyl ethanolamine, an unidentified glycolipid and unidentified lipids (L1,2). Major hopanoids of strain JC90^T were determined to be bacterial hopane derivatives (BHD1,2), diplopterol, diploptene and two unidentified hopanoids (UH1,2). The predominant isoprenoid quinone was identified as menaquinone (MK-6). Anteiso-C_15:0 was determined to be the predominant fatty acid and significant proportions of iso-C_14:0, C_14:0, iso-C_15:0, C_16:0, iso-C_16:0, iso-C_17:0, anteiso-C_17:0 and C_18:02OH were also detected. The results of physiological and biochemical tests support the molecular evidence and allowed a clear phenotypic differentiation of strain JC90^T from all other members of the genus Salinicoccus. Strain JC90^T is therefore considered to represent a novel species, for which the name Salinicoccus halitifaciens sp. nov. is proposed. The type strain is JC90^T (=KCTC 13894^T =DSM 25286^T).     

Grass maturity effects on cattle fed silage-based diets. 1. Organic matter digestion,rumen fermentation and nitrogen utilization

Four silages were harvested at approximately one-week intervals from the same timothymeadow fescue sward. Advanced maturity of the herbage was evidenced by increased neutral detergent fibre [409, 497, 579 and 623 g in 1 kg dry matter (DM)] and decreased nitrogen (N; 29.9, using four ruminally and duodenally cannulated young cattle in a 4 × 4 Latin square experiment. On DM basis (g kg⁻¹), the diet comprised grass silage (700), rolled barley (240) and rapeseed meal (60) and it was given at a rate of 70 g DM (kg live weight)^−0.75 per day.Organic matter digestibility decreased in a curvilinear manner (P_{LINEAR (L)} < 0.001, P_{CUBIC (C)} < 0.01) the values being 0.821, 0.816, 0.758 and 0.747 for the diets based on the four silages in the order of harvest date. Rumen pH increased linearly (P_L < 0.05) and ammonia N concentration decreased curvilinearly (P_L < 0.01, P_C < 0.05) as the grass matured. The molar proportion of acetate in the rumen VFA increased (P_L < 0.001) and the proportion of butyrate decreased (P_L < 0.001) with increased grass maturity. The silage harvest date did not affect the proportion of propionate. The changes in rumen fermentation pattern were associated with a decrease (P_L < 0.05) in rumen protozoal number with increasing maturity of grass.N intake decreased significantly (P_L < 0.001, P_C < 0.01) with the maturity of grass from 167.5 to 118.0 g per day, but duodenal non-ammonia N decreased only from 111.3 to 97.3 g per day indicating greater N losses from the rumen with early-cut silages. The efficiency of microbial protein synthesis in the rumen was not affected by the maturity of grass ensiled. Apparent digestibility of N decreased (P_L < 0.001, P_C < 0.01) and the degradability of N in the rumen decreased (P_L < 0.05) as the grass matured.     

Conformational Preferences of Modified Nucleoside N(4)-Acetylcytidine, ac4C Occur at “Wobble” 34th Position in the Anticodon Loop of tRNA

Conformational preferences of modified nucleoside, N(4)-acetylcytidine, ac⁴C have been investigated using quantum chemical semi-empirical RM1 method. Automated geometry optimization using PM3 method along with ab initio methods HF SCF (6-31G**), and density functional theory (DFT; B3LYP/6-31G**) have also been made to compare the salient features. The most stable conformation of N(4)-acetyl group of ac⁴C prefers “proximal” orientation. This conformation is stabilized by intramolecular hydrogen bonding between O(7)···HC(5), O(2)···HC2′, and O4′···HC(6). The “proximal” conformation of N(4)-acetyl group has also been observed in another conformational study of anticodon loop of E. coli elongator tRNA^Met. The solvent accessible surface area (SASA) calculations revealed the role of ac⁴C in anticodon loop. The explicit molecular dynamics simulation study also shows the “proximal” orientation of N(4)-acetyl group. The predicted “proximal” conformation would allow ac⁴C to interact with third base of codon AUG/AUA whereas the ‘distal’ orientation of N(4)-acetyl cytidine side-chain prevents such interactions. Single point energy calculation studies of various models of anticodon–codon bases revealed that the models ac⁴C₍₃₄₎(Proximal):G₃, and ac⁴C₍₃₄₎(Proximal):A₃ are energetically more stable as compared to models ac⁴C₍₃₄₎(Distal):G₃, and ac⁴C₍₃₄₎(Distal):A₃, respectively. MEPs calculations showed the unique potential tunnels between the hydrogen bond donor–acceptor atoms of ac⁴C₍₃₄₎(Proximal):G₃/A₃ base pairs suggesting role of ac⁴C in recognition of third letter of codons AUG/AUA. The “distal” conformation of ac⁴C might prevent misreading of AUA codon. Hence, this study could be useful to understand the role of ac⁴C in the tertiary structure folding of tRNA as well as in the proper recognition of codons during protein biosynthesis process.     

Analysis of binding curves in multivalent antigen-heterogeneous antibody systems

The binding of antibodies to N-valent antigens can be utilized to gain information on the antibody affinity distribution, under the assumption, that the formation of each bond occurs as an independent event. The analysis of the most widely used plots of binding data (double reciprocal, Scatchard, Sips and the so-called “avidity” plot) leads to expressions which correlate asymptotical features of the binding curves to the antibody site concentration to the antigen valence and to the affinity moments <K^?1>, <K> and <K²>. Only in the “avidity” plot is the shape of the curve independent of the valence of the antigen, depending merely on the antibody affinity distribution.     

Active center and subunit structure of transaldolase from Candida utilis.

Crystalline transaldolase (type III) isolated from Candida utilis is composed of two identical subunits, as shown by the following lines of evidence. 1. Tryptic digestion of the performic acid oxidized enzyme yields the number of ninhydrin- and arginine-positive peptides expected for identical subunits. 2. All attempts to separate both subunits by molecular weight or charge differences have failed. 3. Cyanogen bromide cleavage and sodium dodecyl sulfate gel electrophoresis of S-carboxymethylated transaldolase revealed four distinct peptides designated C₂ to C₅ according to their decreasing molecular weight and one additional peak, C₁, in low yield, presumably an aggregate or partially degraded peptide.By chromatography on Sephadex G-100 the maleylated cyanogen bromide digest from ¹⁴C-labeled β-giyceryl-transaldolase could be separated into four peptide peaks which have been analyzed for their amino acid composition. The largest peptide C₂ with a molecular weight of 16,800 was identified as the active site containing fragment. The four fragments together account for all amino acid residues in the entire protein.From transaldolase (type I) containing four methionine residues three cyanogen bromide peptides could be identified. By addition of the individual peptides a molecular weight of 37,100 ± 3500 could be calculated, which is half the molecular weight of the native enzyme. From experimental data presented so far both isoenzymes of transaldolase can be regarded as “half-of-the-sites” enzymes.     

P.M.R studies on fully methylated aldohexopyranosides and their 6-deoxy analogues using lanthanide shift reagents

The complexes of lanthanide shift reagents (LSR) with permethylated aldo-hexopyranosides and their 6-deoxy analogues having the gluco, galacto, and manno configurations have been studied. On the basis of shift data from Eu(fod)₃ and Pr(fod)₃, and broadening data from Gd(fod)₃, it was found that the LSR bind preferentially to two neighbouring MeO-oxygens having the axial-equatorial relationship. Because of its steric requirements, the C-5 substituent hinders the binding increasingly in the following order: O-2(ax)-O-3(eq)<O-1(ax)-O-2(eq)<O-4(ax)-O-3(eq). Equatorial groups bind the LSR only weakly. Strong binding to O-6 was found when MeO-6 is predominantly “axially” oriented; when this group has the “equatorial” position, O-6 is not favoured over any other equatorial oxygen. In view of the preference of the LSR to bind to an O(ax)-O(eq) site, it is proposed that O-5 is involved in the binding to the axial O-6. Eu(fod)₃ seems to have less tendency to bind to the O-6(ax)-O-5 site than the other two LSR.     

Reassociation and dissociation of cytoplasmic membrane-associated DNA

The relationship between nuclear DNA and cytoplasmic membrane-associated DNA, extracted from a human lymphocyte cell line, was examined by DNA-DNA reannealing and by dissociation of renatured molecules. Up to 2% of the total cellular DNA is found in the cytoplasm as cytoplasmic membrane-associated DNA and of this 2%, approximately 70% is comprised of repeated sequences. These sequences are homologous to only about 4% of the repeated sequences of nuclear DNA. The repeat fraction of cytoplasmic membrane-associated DNA consists of sequences which are only moderately repeated. The number of copies in the average “family” could range from about 1500 copies to as few as 25 copies. A small rapidly reannealing portion of cytoplasmic membrane-associated DNA (C₀t < 4 × 10^?3) appears to consist of sequences derived from a single “family”.About 30% of cytoplasmic membrane-associated DNA reassociates slowly with a $\text{C}{}_0\text{t}\text{1}\text{2}$ value of 223 (unique cytoplasmic membrane-associated DNA). This fraction has homology with about 11% of the unique sequences of nuclear DNA. However, unique cytoplasmic membrane-associated DNA comprises only about 0·6% of the total cellular DNA. If it is assumed that each cell has the same amount of cytoplasmic membrane-associated DNA, homology with 11% of the unique sequences of nuclear DNA suggests that different cells may have different unique nucleotide sequences in the cytoplasm.     

An ecological indicator to evaluate the effect of chemical insecticide pollution management on complex ecosystems

The extensive input of chemical insecticides for pest control is considered as a serious risk to the environment, and the ecological disturbance of chemical insecticides has both positive and negative effects on complex agro-ecosystems. This paper proposed an indicator based on ecological two-sidedness theory and Shannon entropy, which is intended for analyzing informational complexity in a decision network of the chemical insecticide pollution management. The results indicated that the order of the value of R_CC/CP index (where the R_CC/CP index matrix W_CC/CP is defined as the index optimization matrix of comprehensive cost divided by the index optimization matrix of comprehensive profit) for three insect pest-controlling strategies in scallion fields was “applying frequency vibration lamps and environment-friendly insecticides 8 times” (0.8714) < “applying trap devices and environment-friendly insecticides 9 times” (0.8858) < “applying common insecticides 15 times” (0.9077). The treatment “applying frequency vibration lamps and environment-friendly insecticides 8 times” was recognized as the optimal strategy for chemical insecticide pollution management in scallion fields in Shanghai, China. The results demonstrate that our proposed ecological indicator might arouse the interest of policy makers and eco-environmentalists who seek to minimize the use of chemicals, and the farmers who hope to optimize pest-controlling strategies in practice.     

Acyl coenzyme A dehydrogenases and electron-transferring flavoprotein from beef hart mitochondria.

Electron-transferring flavoprotein (ETF) and long-chain acyl coenzyme A (CoA) dehydrogenase (LC-AD) have been purified essentially to homogeneity from beef heart (BH) mitochondria and partially characterized. The spectra of the major yellow acyl CoA dehydrogenase from BH mitochondria, both oxidized and when bleached with C₁₆CoA, were found to resemble those of pig liver (PL) LC-AD. The subunit molecular weight was found to be about 38,000 both by Na-dodecyl sulfate gel electrophoresis and by minimal molecular weight based on flavin content (A₄₅₀, ? = 11.3 × 10³ cm^?1m^?1). The enzyme is probably a tetramer with no interchain disulfide bonds. When assayed in the presence of ETF, relative activities are C₈CoA > C₁₆CoA ? C₄CoA. These findings show that physicochemical and specificity characteristics do not coincide in the pig liver and the beef heart enzymes. The BH ETF is similar to the PL ETF in its spectra, in subunit molecular weight determined by minimal molecular weight (based on flavin content as A₄₃₈), by Na-dodecyl-SO₄ gel electrophoresis, the absence of interchain disulfide bonds, V?_p, and the presence of two subunits/molecule. There were some changes in the amino acid composition concomitant with a decrease in apparent isoelectric point. The pig and beef enzymes were reactive with each other. The turnover number of the beef heart system at “saturating” ETF was 100 mol of 1, 6-dichlorophenol indophenol reduced/min/ mol of LC-AD. Abnormally low activity at low ETF concentrations as compared to high ETF concentrations was seen with the beef heart enzymes as with the pig liver system previously studied and again a material obtained during purification of the ETF similar to the “factor” from pig liver (based on chromatographie and disc-gel electrophoretic behavior) stimulated the low activity, while the high-ETF activity was relatively unaffected, permitting linear double-reciprocal plots over wide ranges of ETF concentration. Fatty-acid-free bovine serum albumin (BSA-FAF) could mimic this effect at equivalent protein concentrations (50–100 μg), as could increased LC-AD concentration and, to a lesser extent, limited aging. Studies of activity at very high concentrations of C₁₆CoA revealed a marked high-substrate inhibition with activity peaking at about 4 μm, the reported critical micelle concentration for C₁₆CoA. The addition of BSA-FAF resulted in more “normal” v vs [S] curves, and although the substrate inhibition was still present it was less severe. The K_m for C₁₆CoA in the presence of BSA-FAF is about 1 μm. These results suggest that the inhibitory species may be the C₁₆CoA micelle, and the BSA-FAF may reverse or alleviate the inhibition by binding acyl CoA in a manner analogous to its binding of fatty acid anions.     

Seasonal variations in bulk tissue, fatty acid and monosaccharide δC values of leaves from mesotrophic grassland plant communities under different grazing managements

Leaves of 26 grass, herb, shrub and tree species were collected from mesotrophic grasslands to assess natural variability in bulk, fatty acid and monosaccharide δ¹³C values under different grazing management (cattle- or deer-grazed) on three sample dates (May, July and October) such that interspecific and spatiotemporal variations in whole leaf tissues and compound-specific δ¹³C values could be determined. The total mean leaf bulk δ¹³C value for plants was −28.9‰ with a range of values spanning 7.5‰. Significant interspecific variation between bulk leaf δ¹³C values was only determined in October (P = <0.001) when δ¹³C values of the leaf tissues from both sites was on average 1.5‰ depleted compared to during July and May. Samples from May were significantly different between fields (P = 0.03) indicating an effect from deer- or cattle-grazing in young leaves. The average individual monosaccharide δ¹³C value was 0.8‰ higher compared with whole leaf tissues. Monosaccharides were the most abundant components of leaf biomass, i.e. arabinose, xylose, mannose, galactose and glucose, and therefore, fluctuations in their individual δ¹³C values had a major influence on bulk δ¹³C values. An average depletion of ca. 1‰ in the bulk δ¹³C values of leaves from the deer-grazed field compared to the cattle-grazed field could be explained by a general depletion of 1.1‰ in glucose δ¹³C values, as glucose constituted >50% total leaf monosaccharides. In October, δ¹³C values of all monosaccharides varied between species, with significant variation in δ¹³C values of mannose and glucose in July, and mannose in May. This provided an explanation for the noted variability in the tissue bulk δ¹³C values observed in October 1999. The fatty acids C_16:0, C_18:2 and C_18:3 were highly abundant in all plant species. Fatty acid δ¹³C values were lower than those of bulk leaf tissues; average values of −37.4‰ (C_16:0), −37.0‰ (C_18:2) and −36.5‰ (C_18:3) were determined. There was significant interspecific variation in the δ¹³C values of all individual fatty acids during October and July, but only for C_18:2 in May (P = <0.05). This indicated that seasonal trends observed in the δ¹³C values of individual fatty acids were inherited from the isotopic composition of primary photosynthate. However, although wide diversity in δ¹³C values of grassland plants ascribed to grazing management, interspecific and spatiotemporal influences was revealed, significant trends (P = <0.0001) for fatty acid and monosaccharide δ¹³C values: δ¹³C_16:0 < δ¹³C_18:2 < δ¹³C_18:3 and δ¹³C_arabinose > δ¹³C_xylose > δ¹³C_glucose > δ¹³C_galactose, respectively, previously described, appear consistent across a wide range of species at different times of the year in fields under different grazing regimes.     

A Single Column Packing for The Gas-Liquid Chromatographic Separation of Various Biologically Important Compounds

The use of a single, commercially available column packing, Tabsorb_R, is described for the g. l. c. separation of a large number of different compounds. The resolution of the homologous members of the following series of compounds was achieved: (1) saturated fatty acids (C₁-C₁₈), (2) normal aliphatic saturated dlcarboxylic acids (C₂-C₁₄), (3) normal aliphatic saturated alcohols (C₁-C₂₄), (4) normal aliphatic saturated amines (C₁C₁₂), (5) the common amino acids except arginlne, histidlne and cysteine, (6) aliphatic hydrocarbons (C₁₀-C₂₀) and (7) monosaccharides. It should be noted that twenty-two monosaccharides including three hexosamines and two anhydrohexoses, could be resolved as aldltol acetates In a single run. In addition, galacturonic, glucuronic and lduronlc acids could be separated from one another as their 1, 4-lactones. The resolution achieved in these series of compounds was found to be consistent and highly reproducible. It is of further interest that certain Isomers of the higher fatty acids and hydrocarbons with one double bond could also be separated from the normal and saturated compounds, respectively. The applicability of “Tabsorb” for the g. l. c. separation, although noted above to be considerably broad, is by far not yet exhausted. These procedures which form the basis for the quantitative determinations of the various compounds studied as demonstrated by analysis of glycopeptldes for neutral hexoses and proteins for the amino acids, can readily be adapted to preparative methods. From the biochemical point of view “Tab-sorb” is an extremely versatile column packing in that it can be used for the identification of many of the common building blocks of natural products.     

1H-nmr study on the preferred conformations of chiral pyridine-dinucleotide models

The synthesis of four chiral NAD⁺ models 1 and their 1,4-dihydro analogs 2 is described. From the temperature dependence of the ¹H-nmr spectra it is concluded that for these compounds two preferred conformations I and II, differing slightly in energy, exist. Both conformations are “folded” with the more or less parallel p-anisyl and pyridine groups mutually gauche, but in I the pyridine group is rotated by about 180° as compared with II, thus leading to a conspicuous difference in orientation of the substituent Z (NH₂CO, C₆H₅NHSO₂, (CH₂)₄NSO₂, or (C₄H₈ON)SO₂) in the pyridine ring toward the anisyl group. The most stable conformation (I) has Z closest to the center of the p-anisyl group. In 360-MHz spectra of the dihydropyridines at low temperature (?10°C), slow interconversion of I and II leads to the observation of an XY pattern for the C-4 methylene protons of the 1,4-dihydropyridine system. The anisochronity in this methylene group is caused mainly by the anisotropy of the neighboring p-anisyl group.