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We tested the usefulness of the 5α(H)-stanol/Δ5-sterol ratios obtained by the tetramethylammonium hydroxide (TMAH) thermochemolysis gas chromatography–mass spectrometry (GC–MS) method used to reconstruct redox events in marine sediments as a redox tracer in lake sediments using cores from Lake Suigetsu, Japan. The lake was previously a freshwater lake and is now brackish owing to excavation works conducted during the 17th century. Other redox tracer analyses (e.g., based on farnesol) have shown that C26(nor-24)Δ22/C26(nor-24)Δ5,22 (24-nordehydrocholestanol/24-nordehydrocholesterol), C28(24Me)Δ22/C28(24Me)Δ5,22 (diatomstanol/diatomsterol), and C27Δ22/C27Δ5,22 (22-dehydrocholestanol/22-dehydrocholesterol) ratios have risen (∼0.53) following strong anoxic conditions that formed in Lake Suigetsu (A.D. 1848 and 1935). This study is the first to show that 5α(H)-stanol/Δ5-sterol ratios determined using the TMAH method could be useful as a redox tracer in lacustrine environments. In particular, the ratio of diatomsterol, which is detected in both brackish and fresh water in relatively high abundance, has the potential to be a very useful tracer of redox events. On the other hand, the other 5α(H)-stanol/Δ5-sterol ratios (C27Δ0/C27Δ5 [cholestanol/cholesterol], C28(24Me)Δ0/C28(24Me)Δ5 [campestanol/campesterol], and C29(24Et)Δ0/C29(24Et)Δ5 [sitostanol/sitosterol] ratios) showed no increasing trends, suggesting the influence of terrestrial inputs from degraded products.

Shallow coastal ecosystems are reservoirs of carbon derived from allochthonous organic matter and autochthonous organic matter produced by microalgae and macrophytes. Carbon stored in vegetated coastal ecosystems has attracted broad attention as an important component of carbon sinks. Characterizing the source of carbon in sediments is essential for quantifying the carbon-sequestration function of shallow coastal ecosystems. In this study, we investigated the origins of organic matter using organic biomarkers (lignin phenols, fatty acids, cutin acids, diacids, and ω-hydroxy acids) in surface sediments in a seagrass-dominated lagoon (Furen Lagoon, Japan). Biomarkers derived from allochthonous vascular plants, such as long-chain fatty acids, showed higher concentrations near river mouths. Furthermore, biomarker signals indicated that sedimentary organic carbon originated in large part from degraded allochthonous vascular plants including roots. A Bayesian mixing model using the ratios of syringyl phenols to vanillyl phenols and cinnamyl phenols to vanillyl phenols indicated that up to about 65% of lignin in the sediments was derived from seagrass. This result indicates a substantial contribution of seagrass to the sedimentary lignin pool in an estuarine seagrass meadow. However, the percent contribution of seagrass to the lignin pool varied, with higher values near a tidal inlet and relatively low values near river mouths. Vertical profiles of organic biomarkers varied with the differences in degradability of organic compounds. Specifically, long-chain fatty acids decreased with increasing depth more than the other compounds, suggesting that they degraded more easily. Conversely, we observed a tendency for lignin phenols to be selectively preserved in the vertical sediment profiles. Our results show that sediment organic biomarkers can provide diverse information such as the composition and origins of organic carbon, the contribution of seagrass derived lignin, and the varying degrees of decomposition. This approach should bring new insights to the estimation of carbon in future blue carbon studies.

Thermodynamic studies on protein adsorption onto chromatographic surfaces mainly focus on the molecular level interaction between proteins and ligands. Yet, not much attention is given to the study of polymer grafted ligand architecture effect on thermodynamic parameters, nor to the relation between chromatographic parameters and the directly obtained thermodynamic parameters. These relations are needed in order to confer meaning and to ease future data interpretation of thermodynamic studies of protein adsorption. In this study, the adsorption of bovine serum albumin monomer (BSAm) onto chromatographic surfaces with grafted ligands was studied from a thermodynamic point of view together with chromatographic data. Isothermal titration calorimetry (ITC) results showed that BSAm adsorption is exothermic ( Δ H ¯ a d s Δ H ¯ a d s > 0) when adsorbs onto Toyopearl SuperQ and a conventional resin (Q Sepharose Fast Flow), showing clear differences in the driving forces of adsorption caused by different ligand architectures. In addition, we found a new relation between the salt required for protein elution and the change in adsorption enthalpy ( Δ H ¯ a d s ) directly measured with ITC, intrinsically connecting both adsorption and desorption mechanisms.

In this study, we analyzed 5α(H)-stanol/Δ 5 -sterol ratios in offshore marine sediments using offline tetramethylammonium hydroxide thermochemolysis (TMAH) gas chromatography–mass spectrometry (GC–MS). Data were used to reconstruct redox events recorded in a southern California marine sediment core (Ocean Drilling Program, Leg167, Hole 1017E). The 24-nordehydrocholestanol/24-nordehydrocholesterol and diatomstanol/diatomsterol ratios clearly recorded oxygen minimum zone (OMZ) fluctuations in southern California during the last 40 kyr. In particular, two ratios corresponding to Marine Isotope Stage 3 (MIS3) varied significantly at the OMZ, exhibiting millennial-scale fluctuations related to Dansgaard-Oeschger shifts of warming and cooling intervals. In contrast, other ratios (cholestanol/cholesterol, campestanol/campesterol, sitostanol/sitosterol, and 22-dehydrocholestanol/22-dehydrocholesterol) showed no significant trends, possibly reflecting the influence of stanol-creating organisms. Our results show that 5α(H)-stanol/Δ 5 -sterol ratios obtained using the TMAH GC–MS method are useable tracers of water column and surface sediment redox conditions in southern California marine sediments.

Delignification by chemical pre-treatment improves the anaerobic digestibility of lignocellulosic biomass, but the by-products can potentially inhibit the digestion process. The present study investigates the inhibitory effect of dissolved lignin on three steps of anaerobic digestion: methanogenesis, acidogenesis, and hydrolysis. Dissolved lignin was extracted from a lignin-rich aquatic macrophyte, and anaerobic toxicity tests were performed in batch mode at dissolved lignin concentrations of 0, 0.5, 1.0, 2.5, and 5.0 g L−1 under mesophilic conditions. Minor reductions of up to 15% and 10% against the control were observed in methanogenic and acidogenic activities, respectively, at a dissolved lignin concentration of 5.0 g L−1. By contrast, hydrolysis efficiency dropped by 25% against the control at a dissolved lignin concentration of 1.0 g L−1 and dropped by 35% against the control at a dissolved lignin concentration of 5.0 g L−1. These results suggest that enzymatic hydrolysis of cellulose is the step that is most susceptible to inhibition by dissolved lignin in the anaerobic digestion process.

The composition of organic matter in early Miocene sediment cores obtained from International Ocean Discovery Program Expedition 359 (IODP Exp. 359) was analysed using tetramethylammonium hydroxide (TMAH) thermochemolysis, a method used to determine concentrations of lignin phenols, fatty acids, sterols and hopanoids. These early Miocene sediment cores contain thick sequences (∼100 m) of interbedded light (organic-poor) and dark (organic-rich) sedimentary layers. A total of 49 samples from cores between 704.4 mbsf to 803.95 mbsf (∼20–23 Ma) at Site U1466 (518 m water depth) was selected with particular emphasis placed on the dark layers, although the light layers were also sampled. While organic compounds were below detection limits in the lighter (white) coloured sedimentary layers, the dark layers, with concentrations of organic carbon up to 20%, contained varying concentrations of all analysed compounds. Although sterols were present, hopanoids were more abundant in the dark layers. The contribution of prokaryotic organisms, estimated from the ratio ([hopanols and hopanoic acids]/[sterols + hopanols and hopanoic acids]), indicates that it was prokaryotic organisms rather than eukaryotic organisms that contributed the majority (∼90%) of the organic matter in the dark layers. In conjunction with this record, more negative δ15N values in the darker layers suggest that the prokaryotic organisms were nitrogen-fixing microorganisms (presumably cyanobacteria). The long-to-short fatty acid ratios, together with sterol distributions and the concentration of lignin phenols, suggest the input of terrestrial plant material occurred during a high total organic carbon (TOC) interval (∼770–810 mbsf), which coincides with more negative carbon isotope ratios. Additionally, the long-to-short fatty acid ratio in this interval was significantly higher in the dark layers than the light layers, suggesting that terrestrial environments suitable for vegetation growth expanded during the interval represented by the dark layers. Furthermore, the ratio of cinnamyl lignin phenols to vanillyl lignin phenols (C/V) is relatively high during these intervals. From these biomarker tracers, we speculate that the terrestrial vegetation was dominated by herbaceous plants adapted to frequent sea-level changes.

The effect of light on BPA degradation by an adapted bacterial consortium was investigated. BPA was completely degraded up to 50 mg l −1 , and the degradation followed first-order reaction kinetics both in the light and in the dark. The degradation half-life of BPA when the consortium was grown in presence of light was 21.9, 17.2, and 12.6 h for concentrations of 10, 20, and 50 mg l −1 , respectively; the degradation half-life of BPA in the dark was 13.1, 10.8, and 10.2 h for concentrations of 10, 20, and 50 mg l −1 , respectively. Therefore, light inhibited BPA biodegradation. However, under both conditions, BPA was completely depleted. The bacterial consortium effectively utilised BPA as a growth substrate to sustain a cell yield of 0.95 g g −1 and 0.97 g g −1 in the light and dark, respectively. A total of ten and nine biodegradation intermediates were detected in the light and dark, respectively. Three bacterial metabolic pathways and one photodegradation pathway were proposed to explain their occurrence. This study demonstrated that bacterial consortia may assemble a wide range of catabolic pathways to allow for efficient degradation of BPA, converting BPA to principally bacterial biomass and metabolites exhibiting low or no oestrogenic activity.

We examined the vertical distribution of hopanoid hydrocarbons and perylene in sedimentary sections from Lake Biwa (Japan) and discuss the results with respect to paleoenvironmental information from around the lake region over the last 120 kyr. Moretene concentration was high above the sections where fall-out events of tephra (tephra AT) occurred ca. 30 kyr before present (BP). This feature suggests that the tephra fall-out events created O 2 deficient conditions, resulting in high activity of methane producing bacteria, and of methanotrophic moretene-producing bacteria becoming active above the tephra layer. Five sections at 10–20 kyr, ca. 35 kyr, ca. 70 kyr, ca. 90 kyr and ca.110 kyr BP exhibited peaks in the ratio of 17α, 21β-22R-homohopane to 17β, 21β-22R-homohopane (hopC 31 αβ/ββ). The n -C 23 / n -C 31 alkane ratio in these sections was 0.6 ± 0.1 ( n = 49), very low if compared with typical characteristics of n -alkanes for a Sphagnum peat environment ( n -C 23 / n -C 31 ca. 3.6; Bingham et al., 2010 ). We conclude that Sphagnum peat may not have been an important contributor of n -alkanes over the past 120 kyr. Comparison with previous pollen records for the lake region suggests that high hopC 31 αβ/ββ values may be caused by production of acidic soil from expansion of temperate conifer (e.g. Cryptomeria ) forest under cold and wet climate conditions. The abundance of perylene relative to sediment or total organic carbon (TOC) was high in the sections at ca. 20 kyr, ca. 70 kyr and ca. 90 kyr BP. These sections were approximately in accord with those having a maximum in hopC 31 αβ/ββ. This concordance indicates that the climatic conditions suitable for the production of perylene quinone in soil surrounding the lake coincided with those for a high hopC 31 αβ/ββ ratio. Thus, hopC 31 αβ/ββ and perylene/TOC might be useful as proxies of soil acidity, temperature and humidity.

Aquatic weeds including submerged macrophyte have been excessively propagated and causing environmental issues in freshwater environment of many countries, and the sustainable treatments have been investigated. In the present study, five submerged macrophyte species dominant in Lake Biwa, Japan, Ceratophyllum demersum, Egeria densa, Elodea nuttallii, Potamogeton maackianus and Potamogeton malaianus were used as a substrate for anaerobic digestion to investigate the chemical composition and the anaerobic digestibility. The lignin content of the submerged macrophyte widely ranged from 3.2 to 20.7%-TS depending on species. The lignin of all macrophytes contained 27.2–59.4% of hydroxycinnamic acids, suggesting they are relatively alkali-labile as compared with woody plants. The total CH 4 yield of submerged macrophytes greatly varied from 161.2 to 360.8 mL g-VS −1 depending on species. The CH 4 conversion efficiency of C. demersum , El. nuttallii , Eg. densa , P. maackianus and P. malaianus was 57.1, 61.4, 60.6, 33.9 and 72.2%, respectively. The results showed that C. demersum , El. nuttallii , Eg. densa and P. malaianus are feasible for anaerobic digestion due to the high methane recovery, whereas P. maackianus was not preferable for anaerobic digestion. The present study revealed that the methane recovery of submerged macrophytes is regulated by the lignin content, as well as other lignocellulosic biomass.

The aim of this study is to examine whether the increased expression of phosphorylated c-Jun (p-c-Jun) and phosphorylated c-Jun N-terminal kinase (p-JNK) are significantly associated with neuronal cell death in diabetic rat retinas and retinas exposed to high glucose. Retinas isolated from six adult male Sprague-Dawley rats and six streptozotocin-induced diabetic rats (DM) were cultured in serum-free medium. The explants from non-diabetic controls were cultured in normal-glucose (N) or high-glucose (HG) medium. Furthermore, neurotrophin-4 (NT-4) and Taurine-conjugated ursodeoxycholic acid (TUDCA) were incubated in HG medium. After 7 days, the numbers of regenerating neurites were counted per explant. After counting, the explants were fixed, cryosectioned, and stained by TUNEL, and also immunostained for p-c-Jun and p-JNK. The numbers of TUNEL-positive, p-c-Jun- and p-JNK-immunopositive cells in the GCL were significantly higher and the numbers of regenerating neurites were significantly lower in the HG and the DM groups than in the N groups. In the HG groups supplemented with NT-4 and TUDCA, the numbers of TUNEL-positive, p-c-Jun- and p-JNK-immunopositive cells were significantly lower and the numbers of neurites were significantly higher than in the HG group without NT-4 and TUDCA. Increased expression of p-c-Jun and p-JNK is associated with neuronal cell death in diabetic rat retinas and retinas exposed to high glucose. Neuroprotective effect of TUDCA and NT-4 is correlated with the suppression of p-c-Jun and p-JNK expression. These results provide a better understanding of the neurodegenerative process underlying DR.

Our recent work has shown that the intestinal absorption of insulin can be improved significantly by coadministration of cell-penetrating peptides (CPPs), especially penetratin. However, a relatively high dose of penetratin is required to adequately stimulate the intestinal absorption of insulin. Therefore, in this study, we sought to determine the CPP that most effectively enhanced intestinal insulin absorption. An in situ loop absorption study using 26 penetratin analogues suggested that the chain length, hydrophobicity, and amphipathicity of the CPPs, as well as their basicity, contribute to their absorption-enhancing efficiency. Moreover, a molecular orbital method with self-organizing maps (SOMs) classification suggested that multiple factors, including the molecular weight, basicity, the lowest unoccupied molecular orbital energy, absolute hardness, and chemical potential of CPPs, are associated with their effects on intestinal insulin absorption. Furthermore, the new CPPs proposed by SOM clustering had a marked capacity to interact with insulin, and their ability to enhance insulin absorption was much stronger than that of the original penetratin. Therefore, the peptide sequence that optimally enhances intestinal insulin absorption could be defined by SOM with the molecular orbital method, and our present work emphasizes the utility of such methodologies in the development of effective drug delivery systems. © 2012 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:469–479, 2013

Carbon dioxide (CO2) sequestration in deep aquifers is one of the options for reducing the emission of greenhouse gases to the atmosphere. It is efficient to inject super critical CO2 into an aquifer if the geomechanical stability of the aquifer -caprock system can be maintained during the injection. An axisymmetric horizontal aquifer -caprock system is modeled including hydromechanical coupling in order to study influence of various factors - such as injection rate, permeability and stiffness of the caprock and aquifer - on the stability and especially the integrity of the caprock. Here the local safety factor is considered as an index of caprock integrity. It is shown that the effective stress state of the caprock close to the injection well could become critical for shear failure quite early in the injection. A buildup of gas pressure in the aquifer causes caprock deformation, inducing an increase in deviatoric stress within the caprock. Larger injection rates and lower aquifer permeability amplify such phenomena, increasing the risk of losing caprock integrity. Increased aquifer stiffness, in contrast, restricts the caprock deformation, and lowers the risk.

Purpose To evaluate the microstructure of the inner and outer retina and the visual function after macular hole (MH) surgery using brilliant blue G (BBG) or indocyanine green (ICG) to make the internal limiting membrane (ILM) more visible. Design Comparative, retrospective, interventional case series. Participants Sixty-three eyes of 63 consecutive cases with MH were studied. Thirty-five eyes of 35 cases were treated with BBG between January and August 2011. Twenty-eight eyes of 28 MH cases were treated with ICG from April 2009 through April 2010. Methods Vitrectomy was performed with a 23-gauge system and 0.25 mg/ml BBG or with 0.125% ICG. Main Outcome Measures The best-corrected visual acuity (BCVA) and the microperimetry-determined retinal sensitivity were measured at baseline and at 3 and 6 months after surgery. The length of the defect of the photoreceptor inner segment/outer segment (IS/OS) junction and external limiting membrane (ELM), the central foveal thickness (CFT), and the thickness of the ganglion cell complex (GCC) were measured in the spectral-domain optical coherence tomographic images. Results The average BCVA was significantly better in the BBG group than in the ICG group at 3 months ( P = 0.021) and 6 months ( P = 0.045) after surgery. The mean retinal sensitivity in the BBG group was improved significantly in the central 2° at 3 and 6 months ( P = 0.001 and P = 0.030, respectively), but was not significantly improved in the adjacent 10°. The length of IS/OS junction defect was significantly shorter in the BBG group at 3 months ( P = 0.048), but was not significantly different at 6 months ( P = 0.135). The length of ELM defect and the GCC thickness were not significantly different between the 2 groups at 3 and 6 months. The CFT was significantly thinner in the ICG group than in the BBG group at 3 and 6 months ( P = 0.013 and P = 0.001, respectively). Conclusions The postoperative BCVA and retinal sensitivity in the central 2° were better in eyes after BBG-assisted vitrectomy. The restoration of IS/OS junction was faster in the BBG group, and the CFT was significantly thinner in eyes after ICG. Brilliant blue G may be a better agent than ICG to make the ILM more visible. Financial Disclosure(s) The author(s) have no proprietary or commercial interest in any materials discussed in this article.

The objective of this study was to investigate the effects of particle size reduction and solubilization on biogas production from food waste (FW). To clarify the effects of volatile fatty acids (VFAs) in the digestion process, the relationship between particle size and VFA accumulation was investigated in detail. For this purpose, substrates of various particle sizes were prepared by bead milling to support hydrolysis. Batch anaerobic digestion experiments were carried out using these pretreated substrates at mesophilic temperature for a period of 16 days. The results of pretreatment showed that the mean particle size (MPS) of substrates ground with a bead mill decreased from 0.843 to 0.391 mm, and solubilization accounted for approximately 40% of the total chemical oxygen demand (total COD) for grinding pretreatment by bead milling. Anaerobic digestion batch experiments revealed that MPS reduced by bead milling at 1000 rpm improved methane yield by 28% compared with disposer treatment. Moreover, this may have increased microbial degradation during the VFA production process with increasing total number of revolutions (operation time × revolutions per minute). However, excessive reduction of the particle size of the substrate resulted in VFA accumulation, decreased methane production, and decreased solubilization in the anaerobic digestion process. These results suggest that optimized reduction of the particle size of the substrate in conjunction with optimized microbial growth could improve the methane yield in anaerobic digestion processes.

Temporal changes in paleoproductivity of Lake Biwa (Japan) over the past 32 kyr have been studied by analyzing bulk organic carbon and photosynthetic pigments (chlorins) in the BIW95-5 core. Primary productivity was estimated on the assumption of C/Norg values of 8 for autochthonous organic matter (OM) and 25 for allochthonous OM and using an equation developed for the marine environment. The estimate indicates that primary productivity ranges from 50 to 90 g C m−2 yr−1 in the Holocene, while it is ∼60 g C m−2 yr−1 on average in the last glacial. Pheophytin a and pheophorbide a are the major chlorins. A downcore profile of chlorin concentration normalized to autochthonous organic carbon (OC) shows a decreasing trend. Chlorin productivity was corrected by removal of the effect of post-burial chlorin degradation. The temporal profile of chlorin productivity thereby obtained resembles that from autochthonous OC. The difference in primary productivity between the Holocene and the glacial for the lake is markedly smaller than that for Lake Baikal situated in the boreal zone. This difference between the two lakes is probably caused by the difference in their climatic conditions, such as temperature and precipitation. Precipitation at Lake Biwa is relatively large during the glacial and the Holocene because of the continuous influence of the East Asian monsoon. Lake Baikal precipitation is generally small as a result of control by the continental (Siberia) climate regime. In addition, a significant difference in productivity between the glacial and the Holocene for Lake Baikal may be essentially controlled by the hydrodynamic systems in the lake. Lake Biwa terrigenous OM input events occurred at least five times over the period 11–32 kyr BP, suggesting enhanced monsoon activity. Molecular examination of the layer with a large input of terrigenous OM during the Younger Dryas indicates that concentrations of terrigenous biomarkers such as n-C27–C31 alkanes, lignin phenols, cutin acids, ω-hydroxy acids and C29 sterols are high, suggesting that soil OM with peat-like material entered the lake as a result of flooding. An enhanced sedimentation rate in the last 3000 years might have been partially caused by agricultural activity around the lake.

The retention and peak spreading in linear gradient elution of charged large biomolecules were investigated by using numerical simulations. Oligo-DNA separation by monolithic anion-exchange chromatography was chosen as a model system. The peak width and the retention were well predicted by using the parameters obtained by gradient elution experiments at different gradient slopes. As the distribution coefficient at the peak retention volume K(R) decreases with increasing molecular size, the peak became sharper for larger DNAs. This is due to very large effective charge (binding site) values of large DNAs (20-60). The peak width was well correlated with K(R) based on the model equation developed for linear gradient elution of proteins. It was shown that the monolithic disk is best suited for very large charged biomolecule separations at high flow velocities with shallow gradients slopes.

Our previous study has shown that there is a good correlation between the number of charges of DNA (from trimer to 50-mer) and the number of binding sites B in electrostatic interaction chromatography (ion-exchange chromatography, IEC). It was also found that high salt (NaCl) concentration is needed to elute large DNAs (>0.6 M). In this paper we further performed experiments with large DNAs (up to 95-mer polyT and polyA) and charged liposome particles of different sizes (ca. 30, 50 and 100 nm) with a monolithic anion-exchange disk in order to understand the binding and elution mechanism of very large charged biomolecules or particles. The peak salt (NaCl) concentration increased with increasing DNA length. However, above 50-mer DNAs the value did not increase significantly with DNA length (ca. 0.65–0.70 M). For liposome particles of different sizes the peak salt concentration (ca. 0.62 M) was similar and slightly lower than that for large DNAs (ca. 0.65–0.70 M). The binding site values (ca. 25–30) are smaller than those for large DNAs. When arginine was used as a mobile phase modulator, the elution position of polyA and polyT became very close whereas in NaCl gradient elution polyT appeared after polyA eluted. This was mainly due to suppression of hydrophobic interaction by arginine.

We characterized the compositions of organic compounds in a Cheremushka bog sediment core (deposited over the last 35 kyr), located at the eastern coast of Lake Baikal, to obtain basic information about the terrestrial organic matter (OM) which contributed to Lake Baikal sediments. The bog sediment was analyzed for the molecular composition of n -alkanes, lignin phenols and n -C 24 to C 30 alkanoic acids, as well as the carbon isotopic composition of plant wax derived n -C 27 to C 33 alkanes. Concentrations of lignin phenols [vanillyl (V) plus syringyl (S) phenols] normalized to total organic carbon (TOC) in the Holocene are twice those for the last glacial maximum (LGM), while concentrations of TOC-normalized n -C 24 to C 30 alkanoic acids do not change markedly in this period. Thus, the ratio of lignin phenols to n -C 24 to C 30 alkanoic acids increases from the LGM to the Holocene. This result is essentially consistent with pollen analysis indicating an expansion of woody plants in the Holocene and a prevailing herb-abundant environment for the LGM. The δ 13 C values of n -C 27 to C 33 alkanes (e.g. −29‰ to −33‰ for C 31 ) indicate the presence of C 3 -dominant plants throughout the core. The contribution of terrestrial OM to Lake Baikal sediments was estimated using the biomarkers, on the assumption that the OM in the bog sediments is a representative of the terrestrial OM around the lake. Hence, the estimation using lignin phenol or n -C 24 to C 30 alkanoic acid parameters indicates that 11–24% of the TOC in the Academician Ridge sediments is land-derived for both the Holocene and the LGM, which is similar to the estimates from C/N values of bulk OM. However, the estimates for terrestrial OM using the n -C 27 to C 33 alkane parameter are generally higher than those using lignin phenol or n -C 24 to C 30 alkanoic acid parameters. The difference is thought to be associated with the difference in source and behavior of these biomarkers.

As it is important to understand how protein conformational changes affect the separation performance in ion exchange chromatography (IEC), we investigated two model systems, unfolded proteins (lysozyme and bovine serum albumin) with urea and dithiothreitol, and PEGylated proteins (lysozyme attached with polyethyleneglycol molecular weight 5000). Linear gradient elution IEC experiments were carried out and the data were analysed by our model previously presented in order to obtain the binding site value B and the peak salt concentration IR. Unfolded proteins (bovine serum albumin and lysozyme) with urea and dithiothreitol showed weaker retention and larger binding site values compared with the values for native proteins. Multiple PEGylated lysozyme peaks were separated, and eluted earlier than the native peak appeared. There is a good correlation between B and IR for PEGylated lysozymes.

We applied the model described in our previous paper to the rapid scale-up in the ion exchange chromatography of proteins, in which linear flow velocity, column length and gradient slope were changed. We carried out linear gradient elution experiments, and obtained data for the peak salt concentration and peak width. From these data, the plate height (HETP) was calculated as a function of the mobile phase velocity and iso-resolution curve (the separation time and elution volume relationship for the same resolution) was calculated. The scale-up chromatography conditions were determined by the iso-resolution curve. The scale-up of the linear gradient elution from 5 to 100mL and 2.5L column sizes was performed both by the separation of beta-lactoglobulin A and beta-lactoglobulin B with anion-exchange chromatography and by the purification of a recombinant protein with cation-exchange chromatography. Resolution, recovery and purity were examined in order to verify the proposed method.

A range of conifers of the southern hemisphere have been analysed (using gas chromatography–mass spectrometry (GC–MS)) for oxygenated cyclic diterpenoids. Parts of the trees examined include resins, cones, shoots, woods and barks. Trimethylsilyl (TMS) derivatives were mainly used for identification purposes. Identifications were facilitated by using previous work based on classical isolation procedures on certain species. Rules for interpreting the mass spectra of such compounds in the derivatised form were derived and are described. A compilation of such compounds with mass spectra and retention times now exists. Trends relating to the part of the tree analysed and to the tree family/genus were observed as follows: (a) labdanoids were confined to resin samples with two exceptions (Podocarpus falcatus and Podocarpus elongatus shoots); (b) all resin samples examined contained labdanoids of some kind; (c) ferruginol derivatives dominated the phenolic compounds in the Cupressaceae; (d) callitrisic acid was only found in Callitris sp.; (e) Podocarpus samples were dominated by totarol derivatives (particularly) and structures related to ferruginol; (f) other members of the Podocarpaceae were dominated by quite different compound types. A Microcachrys sp. uniquely contained an unusual and complex mixture of alcohols/ketones/aldehydes of the pimarane/abietane types; (g) Phyllocladus species contained only ferruginol structures with no evidence for totarol types; and (h) Araucaria bidwillii was the only species in which clerodanes could be found despite the fact that several Araucaria and Agathis species were analysed. Three new compounds of the totarol class have been identified and two of the ferruginol class. A review and classification (including principal stereochemical features) of di- and tricyclic oxygenated diterpenoid compounds present in members of the Araucariaceae, Cupressaceae, Phyllocladaceae, Pinaceae and Podocarpaceae based on this and previous work are presented. Distinctions between the families may be made on the basis of (a) a strong preference for C-19 oxygenation, rather than C-18 oxygenation, in the Cupressaceae and Podocarpaceae; (b) the apparently unique occurrence of the ent-clerodane and ent-labdane classes of diterpenoids in certain species of the Araucariaceae; (c) certain highly oxygenated ferruginols are unique to the Podocarpaceae and Cupressaceae; and (d) podocarpic acid has only been found unequivocally in the Podocarpaceae.

This investigation attempts to examine the hydrophobic interactions of ion-exchanger's ligand on protein adsorption at mild to high ionic strengths in solution. Adsorption isotherms and enthalpies of lysozyme and myoglobin on cation exchanger resin (CM-Sepharose, SP-Sepharose and SOURCE 30S) were measured at selected ionic strengths of phosphate buffer. Binding enthalpies showed that adsorption occurs mainly by hydrophobic interaction at high ionic strength. The investigation found the binding affinities and enthalpies were ligand-dependent and CM-Sepharose contains the most hydrophobic ligand among those studied. Results of this study provide further insight into the hydrophobic effect of ion-exchange resin on chromatography applications for protein purification under high ionic strength conditions.

Hydroxyapatite (HA) chromatography has been used extensively for the separation of single-strand deoxyribonucleic acids (ssDNA) and double-strand DNA (dsDNA). However, the details of the mechanism of ssDNA and dsDNA separation by hydroxyapatite are still not clear. In this study, we examined the effects of binding parameters (i.e. salt concentration, temperature and pH) and the effects of ssDNA and dsDNA base composition and sequence (i.e. GC content, length, secondary structure effects and GC or CG stacking rich, …) on the binding behavior. The affinities and binding amounts were obtained from equilibrium batch isotherm analysis, and the binding enthalpies were derived from isothermal titration calorimetry (ITC). Isotherm analysis reveals that electrostatic forces are the main driving force of dsDNA binding to HA. Due to hydrophobic bases and the negative charge of the phosphate backbone, the ssDNA molecule binds to HA with both hydrophobic and electrostatic interactions. Through thermodynamic analysis, we find that the adsorption enthalpies of both ssDNA and dsDNA are endothermic under all of the conditions studied and the dehydration step in the binding process plays a key role. The binding behavior responses to the binding parameters are discussed accordingly.

Linear gradient elution experiments were carried out on monolithic anion-exchange chromatography (AEC) with oligo-DNAs of various sizes (4–50mer, molecular weight M W = 1200–15,000) and compositions in order to investigate the retention mechanism. The binding site ( B ) values as well as the peak salt elution concentration I R values were determined. The B values determined for the monolithic AEC were similar to the values for non-porous AEC and porous AEC. The B value increased linearly with the number of charges (bases) of single-strand DNA when M W is less than ca. 3600 (12mer). When M W is greater than 6000, the slope of B versus M W decreased, and became very small at M W > 30,000. The I R value also increased linearly with M W for M W M W for M W > 10,000. It was shown that a very difficult separation of a single-strand 50mer poly(T) and a double-strand 50mer poly(A) and poly(T) was accomplished within 10 min by using a very shallow gradient at a high initial salt concentration (0.5 M) and a high flow-velocity (2.7 cm/min).

A 10 m sediment core from Academician Ridge in Lake Baikal was analyzed for its molecular composition using on-line TMAH (tetramethylammomium hydroxide) thermochemolysis. Major products are lignin phenols, n-C14 to C30 fatty acids (alkanoic acids), cutin acids, hydroxy acids and aliphatic dicarboxylic acids. Lignin phenols are abundant in warmer periods (the interglacial: marine isotope stage (MIS) 5e and MIS 1), but extremely low in the other (colder) periods. This result coincides well with pollen records reported for a core near the present site, where an expansion of coniferous forests in sub-stage 5e and MIS 1 was implied. Normal C24–C30 alkanoic acids, important components of plant wax esters, are abundant in 5e and MIS 1 and are present in significant amounts in the other (colder) periods, unlike the lignin phenols. A high abundance of n-C24 to C30 alkanoic acids relative to lignin phenols in the Bolling–Allerod warm period suggests an enhanced development of herbs. It is implied from comparison of the sedimentary lignin phenol record with fossil pollen records and lignin phenol analysis of modern pollen that the ratio of cinnamyl phenols to vanillyl phenols may serve as an indicator of pollen contribution to sedimentary organic matter.

The biomarker compositions of six ambers (two glessites, two succinites, goitschite, and stantienite) from the Upper Oligocene of the Bitterfeld deposit in Germany were analyzed by gas chromatography–mass spectrometry (GC-MS) to determine their botanical origin. The extracts of the two glessites have similar compositions comprised of sesquiterpenoids of the cadinane and bisabolane classes and triterpenoids of the oleanane, ursane, lupane, and allobetulane classes. These triterpenoids are common in angiosperms. The botanical origin of the glessite ambers is proposed to be Betula, because allobetulane triterpenoids are the specific biomarkers for that genus. The constituents of two succinites and goitschite are similar, mainly consisting of succinic acid, borneol monoterpenoids, cadinane sesquiterpenoids, isopimarane, pimarane, and abietane type diterpenoids, with a predominance of dehydroabietic acid. Monoterpanyl succinates and monoterpanyl diterpenoates are also present. Monoterpanyl esters are one notable characteristic of succinite type ambers. The terpenoid compositions of these ambers indicate Pinaceae as the botanical source. Furthermore, the detailed examination of our results suggests that the ambers are derived from resins of Pinus or Picea (Pinaceae). The stantienite yielded an unresolved complex mixture (UCM) of branched and cyclic compounds, C12–C32 methyl alkanoates (maximizing at C25) and C12–C27 dimethyl alkanedioates (maximizing at C18), with minor amounts of aromatic diterpenoids and triterpenoids including hopanes. The series of methyl esters are regarded to be derived from leaf waxes and suberins of higher plants. The carbon preference indices (CPI) of the methyl alkanoates and alkanedioates indicate full diagenetic/catagenetic maturity of the amber. Unknown compounds A and B (MW = 400) are major components in the amber and also occur in the Miocene resin of Cupressospermum saxonicum. Thus the botanical source of stantienite may be in part the same as the Miocene resin. The amber is also admixed with microbial triterpenoids, leaf wax and biopolymer products indicating mixed sources.

Our simple method for optimization of the elution salt concentration in stepwise elution was applied to the actual protein separation system, which involves several difficulties such as detection of the target. As a model separation system, reducing residual protein A by cation-exchange chromatography in human monoclonal antibody (hMab) purification was chosen. We carried out linear gradient elution experiments and obtained the data for the peak salt concentration of hMab and residual protein A, respectively. An enzyme-linked immunosorbent assay was applied to the measurement of the residual protein A. From these data, we calculated the distribution coefficient of the hMab and the residual protein A as a function of salt concentration. The optimal salt concentration of stepwise elution to reduce the residual protein A from the hMab was determined based on the relationship between the distribution coefficient and the salt concentration. Using the optimized condition, we successfully performed the separation, resulting in high recovery of hMab and the elimination of residual protein A.

Although linear gradient elution ion-exchange chromatography is best suited for very fine separation of proteins, it is not commonly employed for the actual protein purification process. Instead, stepwise elution is usually used. However, it is difficult to design an efficient stepwise elution process as small changes in the buffer salt concentration often affect the retention volume and the peak width significantly. In this paper, a design calculation method for stepwise elution proposed previously was modified and extended. This method uses the data obtained by linear gradient elution experiments. The efficiency of the process was examined on the basis of the peak width and the separation time. The advantages using convection-aided chromatography media (monolithic columns) were investigated.

Simple methods for the optimization of ion-exchange chromatography of proteins in our previous papers were applied to cation-exchange chromatography purification of monoclonal antibodies (Mab). We carried out linear gradient elution experiments, and obtained the data for the peak salt concentration and the peak width. From these data, the distribution coefficient as a function of salt concentration, and the height equivalent to a theoretical plate (HETP) as a function of mobile phase velocity were calculated. The optimized linear gradient elution conditions were determined based on the relationship between buffer consumption and separation time. The optimal stepwise elution conditions were determined based on the relationship between the distribution coefficient and the salt concentration.

A stratigraphic study of total organic carbon (TOC) concentration, carbon isotopic composition of total organic matter (TOM), lipids (n-alkanes, n-alkanols, n-alkanoic acids and sterols) and lignin/cutin compounds has been conducted for a Lake Baikal core (323-PC1) over the last glacial/interglacial. TOC concentrations (mg g−1 dry sediment) are low (2–4) in the glacial sections, but high (10–29) in the interglacial ones. The concentrations of land plant-derived compounds such as n-C14 to C28 ω-hydroxyalkanoic acids, lignin phenols (p-hydroxyl, vanillyl and syringyl), total cutin acids, normalized to TOC in the interglacial, are 5–6 times higher than those in the glacial. Carbon isotopic compositions of TOM are −24 ± 0.9‰ in the glacial, while they are −27 ± 1.3‰ in the interglacial. The heavy isotope composition of TOM in the glacial might be due to a contribution from submerged plants and/or grasses. The light isotopic composition of TOM for the interglacial may be due to a contribution from both land plant- and phytoplankton (mainly diatom)-derived OM. Autochthonous organic carbon contents were estimated from the C/N ratio corrected for the effect of inorganic nitrogen and based on the assumption of C/N values of 8 for autochthonous and 25 for allochthonous OM. The estimation indicates that autochthonous OC comprises 74 ± 6% of TOC for the glacial and 72 ± 9% for the interglacial, respectively. Primary palaeoproductivity was estimated to be below 20 gC m−2 y−1 in the glacial (before 18 ka), but high (100–120 gC m−2 y−1) in the interglacial (11–8 ka), which is 10–30% higher than the modern values.

Thermochemolysis with tetramethylammonium hydroxide (TMAH) was conducted for 18 humic acids (HAs) obtained from a wide range of soils, and the yields and compositions of the released compounds were analyzed in terms of the degree of humification and the proportions of C species estimated by 13C CPMAS NMR. Eighty-six peaks could be assigned on GC/MS chromatograms, and notable differences in the kind of compound assigned were not found among the HAs. The total yields of aliphatic compounds ranged from 0.40 to 4.2 mg/g. C12- to C18-even fatty acid methyl esters (FAMEs) accounted for 30–66% of the entire aliphatic compounds assigned, followed by C20- to C34-even FAMEs (4.8–47%), C9- to C33-odd FAMEs (11–26%), and C16- to C30-even dicarboxylic acid dimethyl esters (DAMEs; 0.3–9.1%). The total yields of aliphatic compounds decreased with decreasing gross alkyl C during the progress in the degree of humification, although their proportion in gross alkyl C was small. On the other hand, the total yields of aromatic compounds ranged from 1.7 to 37 mg/g. Of these, 7.5–82% and 0.12–64% were phenol derivatives that were possibly derived from lignin and benzenepolycarboxylic acid polymethyl esters, respectively. The yields of phenol derivatives and stilbene compounds, possibly originating from lignin dimers, decreased with an increase in the degree of humification, contrary to the increase in the proportion of aromatic C in total C. The yields of heterocyclic compounds, namely, indole and proline derivatives, were also higher in HAs with lower degrees of humification. The variation in the yield of benzenepolycarboxylic acid polymethyl esters suggested parallel progression of oxidation with increasing degree of humification. Aromatic compounds responsible for the high aromaticity and the dark color of HAs, such as quinones and polycyclic compounds, were not detected even in HAs with a high degree of humification.

The binary diffusivities of water in low molecular weight sugars; fructose, sucrose and a high molecular weight carbohydrate; maltodextrin (DE 11) and the effective diffusivities of water in mixtures of these sugars (sucrose, glucose, fructose) and maltodextrin (DE 11) were determined using a simplified procedure based on the Regular Regime Approach. The effective diffusivity of these mixtures exhibited both the concentration and molecular weight dependence. Surface stickiness was observed in all samples during desorption, with fructose exhibiting the highest and maltodextrin the lowest.

Purpose: To assess the potential role of monocyte chemotactic protein-1 (MCP-1) in the pathogenesis of proliferative vitreoretinopathy (PVR) and to investigate its possible interaction with the macrophage migration inhibitory factor (MIF). Methods: We assayed MCP-1 and MIF levels in the vitreous samples of 85 consecutive patients with PVR (29 eyes), rhegmatogenous retinal detachment (RRD; 22 eyes), and macular hole or idiopathic epimacular membrane (controls; 34 eyes), by enzyme-linked immunosorbent assay. Results: Vitreous levels of MCP-1 were 1760.7 ± 471.3 pg/mL (mean ± SD) in PVR patients, 1200.4 ± 579.8 pg/mL in RRD patients, and 436.3 ± 286.1 pg/mL in the controls. Vitreous MCP-1 levels in PVR patients were significantly higher than those in RRD patients and in the controls (P < .0001, respectively). MCP-1 levels in grade C of PVR (1883.7 ± 479.5 pg/mL) were significantly greater than those in grade D (1437.8 ± 258.8 pg/mL) (P = .0112). Vitreous concentrations of MCP-1 had no correlation with those of MIF. Conclusions: The results indicate the possibility that MCP-1 may have a role mainly in the early stage of PVR and that the role of MCP-1 in PVR may differ from that of MIF.

As various types of gels or gelled solutions are important in food industries, mass transport phenomena in sugar solutions gelled with polysaccharides such as agar, pectin, sodium alginate and a protein (gelatin) were investigated. Water diffusivities from the isothermal regular regime drying curves, desorption isotherms, viscosities, and electroconductivities were determined for gelled sugar solutions. Gels tested were divided into two types (I and II). Type I gels (agar and sodium alginate) did not affect the transport properties (water diffusivities) of sugar solutions although the viscosities changed very much. The water diffusivities of Type II gels (gelatin and pectin) were much lower than those for Type I gels, especially at low water contents. The desorption isotherms of Type I and Type II gels were similar. Although the electroconductivities decreased with increasing sugar concentration, they did not change when the solutions were gelled. It is suggested that Type I gel network was quite uniform during drying while a rather dense gel network was formed near the surface of Type II gels after a certain period of drying. This kind of hard skin layer might govern the water diffusivity (and the drying rate).

This study examined the interaction mechanisms of beta-lactoglobulins A and B (Lg A, Lg B) with an anion exchanger, Q-Sepharose at pH near the isoelectric point at which the proteins are expected to be electrically neutralized under various NaCl concentrations and temperatures by the equilibrium binding analysis and the adsorption enthalpy directly measured by isothermal titration calorimetry. The data evaluated from isotherms fitted by the Langmuirean model reveal that the addition of NaCl considerably reduced the binding affinities and capacities of both the proteins with Q-Sepharose at pH 5.2, indicating that electrostatic forces are dominant during the adsorption. However, the hydrophobic interaction seems to be involved in adsorption as well at a higher NaCl concentration, and the adsorption enthalpies confirm this suggestion. In addition, the effects of temperature on the equilibrium binding behaviors for Lg A or Lg B with Q-Sepharose were found to be salt concentration-dependent, probably due to their different binding mechanisms at 0.03 M and 0.3 M NaCl. Where, at 0.3 M NaCl, the hydrophobic interaction plays a more pronounced role. This implication was again supported by the adsorption enthalpies. The presented data provide further insight to the interaction mechanisms between proteins and ion exchangers, facilitating the optimization of protein separations.

Six piston cores were collected along the NW–SE line in the Okinawa Trough close to the Chinese continental shelf edge and dated by 23 AMS 14 C measurements, along with planktonic δ 18 O stratigraphy and an ash layer dated as ∼6.3 ka. On a time scale to ∼19 ka, the ratio of total organic carbon to total nitrogen concentration (C/N ratio) displays gradual decreases toward the present and suggests the decrease of vascular plant fragment input to the trough from continental rivers. This suggestion is confirmed by similar decreases of lignin phenol content and ratio of C 28 fatty acid to C 16 fatty acid content in core RN93-PC3. The gradual decrease of land plant fragment input is interpreted to be a consequence of the retreat of the continental coastline during the deglacial period.

Eicosanoid receptors exhibit a highly conserved ERY(C)XXV(I)XXPL sequence in the second intracellular loop. The carboxyl end of this motif contains a bulky hydrophobic amino acid (L,I,V, or F). In human thromboxane A2 receptor (TXA(2)R), phenylalanine 138 is located at the carboxyl end of this highly conserved motif. This study examined the function of the F138 in G protein coupling. F138 was mutated to aspartic acid (D) and tyrosine (Y), respectively. Both mutants F138D and F138Y showed similar ligand binding activity to that of the wild type TXA(2)R. The Kd and Bmax values of either mutant were comparable to those of the wild type receptor. However, both mutants showed significant impairment of agonist induced Ca(2+) signaling and phospholipase C activation. These results suggest that the F138 plays a key role in G protein coupling.

Purpose: To evaluate the properties of the retina of a Japanese patient with enhanced S-cone syndrome by analyzing electroretinograms (ERGs) and visual evoked potentials (VEPs) elicited by different spectral stimuli. Methods: Ganzfeld spectral flashes in the presence of strong white adapting background illumination were used to elicit cone ERGs and VEPs. Results: The cone ERG elicited in the patient by short wavelength stimuli was distinctly different from the normal S-cone ERG. The action spectrum of the cone ERG confirmed its relative hypersensitivity to short wavelengths. The action spectrum of the VEP for the patient showed a similar relative hypersensitivity to short wavelengths. The response of the VEPs to short wavelength stimuli was different in waveform from the VEP response to longer wavelength stimuli observed in a normal subject. Conclusions: These results indicate that the hypersensitivity to short wavelengths is transmitted to the central nervous system and that there is a short wavelength transducing photopigment in many of the photoreceptors, either abnormal S-cones or photopic rods.

The retention and the resolution of β-lactoglobulin A and B (LgA, LgB) were investigated with various ion-exchange chromatography media. The number of sites involved in the retention (adsorption) decreased as the mobile phase pH approached the isoelectric points pI (=5.1–5.2). However, even at pH 5.2 both LgA and LgB were retained on anion- and cation-exchange chromatography columns. The separation (resolution) of LgA and LgB became better when the pH approached the pI in anion-exchange chromatography columns where the number of adsorption site values are small (ca. 2–3). The two proteins were not separated on cation-exchange chromatography columns. Factors affecting the resolution and the retention near the pI were discussed.

Very fine separation of proteins by stepwise elution ion-exchange chromatography is very often a unstable process. To characterize the unstability of such processes the elution volume variations were examined by the model equation which contained the ion-exchange capacity and the number of adsorption sites. The data needed for the model calculation were obtained from gradient elution experiments. As a model separation system stepwise elution of a model protein (beta-lactoglobulin) near the isoelectric point on a weak cation-exchange chromatography column was chosen. The elution volume varied significantly with a small change in the ion-exchange capacity. It was found that the ionic strength of the elution buffer must be adjusted in order to compensate a change in the elution volume due to the ion-exchange capacity variations. The ionic strength and the pH of the elution buffer were also found to be important variables affecting the elution volume. In this model separation system, it was indicated that the pH should be within +/-0.1 unit and the ionic strength within +/-0.002 mol/l in order to meet the criteria (+/-5% elution volume variation). It is recommended that gradient elution data be obtained for predicting elution volume variations in stepwise elution. By using the gradient elution data the process diagnosis can be performed, and the important information on the process stability can be obtained.

Human leukemia (HL) 60 cells were differentiated by dimethylsulfoxide (DMSO) treatment to granulocyte-like cells, leukotriene (LT) synthesizing activity of which was increased in response to the differentiation of the cells. Four synthesizing enzymes, cytosolic phospholipase A2 (cPLA2), 5-lipoxygenase (5-LO), LTA4 hydrolase and LTC4 synthase, and an enzyme associated protein, 5-lipoxygenase activating protein (FLAP) are involved in the generation of LTC4 and LTB4. We examined the expression of messenger RNA (mRNA) for these LT synthesizing enzymes and an associated protein in DMSO differentiated HL-60 cells by reverse transcriptase polymerase chain reaction (RT-PCR). The production of LTC4 and LTB4, measured by radioimmunoassay (RIA), was increased after the incubation with DMSO for more than 3 days. Messenger RNA abundance for 5-LO, LTC4 synthase and LTA4 hydrolase was increased, that for FLAP was stable, but that for cPLA2 was decreased. These results indicate that DMSO induced increase of LT synthesis is associated with the increase of mRNA expression of 5-LO, LTC4 synthase and LTA4 hydrolase, although the precise regulatory mechanisms of the increased mRNA expression are not determined. We also investigated an action of dexamethasone (DEX) on DMSO-induced enhancement of LT synthesis. DEX suppressed DMSO induced increase of LTC4 synthesis, but rather enhanced DMSO induced LTB4 production. The DEX attenuated the DMSO-induced increase of mRNA expression for LTC4 synthase, but showed no effect on that for LTA4 hydrolase. The inhibition of LTC4 synthesis is associated with the suppression of mRNA expression for LTC4 synthase. However, increased LTB4 synthesis by DEX is regulated by the mechanisms which are independent from mRNA level of LTA4 hydrolase.

Cone electroretinograms (ERGs) in response to different color flashes were examined using a Ganzfeld stimulus in 19 eyes after successful retinal detachment surgery. In the operated eyes, the short wavelength sensitive (S-) cone b-wave was reduced more than the mixed long (L-) and middle (M-) wavelength sensitive cone b-waves. The ratio of the S-cone ERG b-wave amplitude between operated eyes and fellow eyes was significantly lower than the L- and M-cone ERG b-waves (P < .01). These ERG results indicated that the S-cone system is more impaired than the L- and M-cone systems after retinal detachment surgery.

Background Cytokines, such as interleukin (IL)-5, produced by T helper type 2 (Th2) cells appear to play an important role in the inflammatory processes associated with atopic dermatitis. The roles of cytokines produced by Th1 cells remain controversial. Objective We examined IL-5 and IL-2 mRNA abundance in and protein production by peripheral blood mononuclear cells (PBMCs) from patients with atopic dermatitis and compared with those from controls. Methods Peripheral blood mononuclear cells were isolated from six children with atopic dermatitis and six control children, and stimulated with both phytohemagglutinin (PHA) and phorbol 12-myristate 13-acetate (PMA). The abundance of IL-5 and IL-2 mRNA in PBMCs was measured by reverse transcription-polymerase chain reaction analysis. The production of IL-5 and IL-2 by PBMCs was also determined by enzyme-linked immunosorbent assay. Results After incubation with PHA and PMA, PBMCs from atopic children showed significantly higher IL-5 mRNA abundance (P .05) and IL-5 production ( P .01) as well as a lower amount of IL-2 mRNA ( P = .056) and IL-2 production (P .05) than those from healthy controls. The time course of changes in IL-5 mRNA abundance induced by PHA and PMA in PBMCs from atopic children differed markedly from that observed with healthy controls, whereas the time course of changes in IL-2 mRNA abundance were similar between the two groups. Conclusions The increased IL-5 and decreased IL-2 production observed with PBMCs from children with atopic dermatitis may underlie the activation of eosinophils and high serum immunoglobulin E concentrations also apparent in such individuals. An imbalance in the number and activity of Th1 and Th2 cells is likely to be responsible for the abnormal pattern of cytokine production in atopic dermatitis.

We examined the action of Shinpi-To (Formula divinita; TJ-85), a granular extract of seven Chinese medicinal herbs that is used in treating childhood asthma, on the leukotriene synthesis in rat basophilic leukemia-2H3 cells (RBL-2H3 cells). IgE-loaded cells were stimulated with anti-IgE serum in the presence or absence of Shinpi-To. Released LTC4 and LTB4 were measured by radioimmunoassay (RIA). Shinpi-To significantly inhibited IgE-mediated synthesis of leukotriene (LT)C4 and LTB4. To identify the inhibitory sites, we investigated the action of this extract on four synthetic enzymes, phospholipase A2 (PLA2), 5-lipoxygenase (5-LO), LTC4 synthase, and LTA4 hydrolase. Shinpi-To inhibited the A23187-stimulated release of [3H]arachidonic acid (AA) from the cell membrane, reflecting an effect on PLA2 activity. It also suppressed production of LTC4 and LTB4 when cell lysates were incubated with AA as substrate. It did not inhibit the production of LTC4 and LTB4 when LTA4-free acid was used as the substrate. Shinpi-To did not inhibit the IgE-mediated increase of intracellular Ca2+ ([Ca2+]i) concentration. Results indicate that Shinpi-To inhibits LT synthesis by inhibiting PLA2 and 5-LO activities without affecting the mobilization of [Ca2+]i.