Protein-protein interaction conferring stability to an extracellular acetyl (xylan) esterase produced by Termitomyces clypeatus

Acetyl esterase (AE) activity present in the culture filtrate of Termitomyces clypeatus was separated into lower molar mass (LMM) and higher molar mass (HMM) protein fractions during BioGel P-200 gel chromatography. AE was purified as a 30 kDa nonglycosylated protein from LMM fractions by CM-Sepharose ion exchange chromatography and HPGPLC. Although the HMM fraction had a number of enzyme activities (sucrase, beta-xylosidase, beta-glucosidase, and alpha-L-arabinofuranosidase) other than AE, protein present in the fraction was eluted as a single protein peak in HPGPLC and gave a single band in native PAGE. The fraction, subsequently purified by DEAE-Sephadex chromatography, was a SDS-PAGE homogeneous 80 kDa glycoprotein, but with both AE and cellobiase activities. The aggregate dissociated during ConA-Sepharose chromatography and 30 kDa AE and 56 kDa glycosylated cellobiase were purified separately. The dissociation caused significant loss of cellobiase activity but not that of AE. AE purified from both HMM and LMM fractions was characterized to be the same enzyme in terms of molar masses, pI (7.3), and other physicochemical properties. AE as an aggregate with cellobiase showed higher thermostability, temperature optimum, and resistance toward chemical denaturants than those of purified AE. Compared to cellobiase purified earlier from the same fungus, the enzyme present with AE in the aggregate also showed higher catalytic activity, thermostability, and temperature optimum. The study indicated that the formation of such SDS-resistant enzyme aggregate was associated with significant changes in the physicochemical properties of the enzymes, mainly toward improvement of rigidity of enzymes, and sometimes with the improvement of catalytic activity.     

Inhibitory Activity by Barley Coffee Components Towards Streptococcus Mutans Biofilm

It was shown that barley coffee (BC) interferes with Streptococcus mutans adsorption to hydroxyapatite. After BC component fractionation by dialysis and gel filtration chromatography (GFC), it was found that the low molecular mass (<1,000 Da) fraction (LMM fraction) containing polyphenols, zinc and fluoride ions and, above all, a high molecular mass (HMM > 1,000 kDa) melanoidin fraction display strong anti-adhesive properties towards S. mutans. In this study, we have further examined the potential of BC, BC LMM fraction and BC HMM melanoidin fraction as caries controlling agents by evaluating their anti-biofilm activity.The effects of BC and BC fractions on biofilm formation by S. mutans ATCC 25175 and its detachment from pre-developed biofilms were evaluated by microtiter plate assay. It was found that BC and its fractions, at concentrations ranging from 60 to 15 mg ml⁻¹ that are devoid of antimicrobial activity, inhibited S. mutans biofilm formation. An increase of S. mutans ATCC 25175 detachment from 24 h developed biofilm was observed at the highest tested concentrations. Interestingly, BC and BC fractions also showed anti-biofilm activity towards a variety of S. mutans clinical strains isolated from saliva, plaque and caries lesions of adult donors. In general, the HMM melanoidin fraction was more active than the LMM fraction. These findings, classifying BC LMM fraction and BC HMM melanoidin fractions as natural anti-biofilm agents, represent the basis for studying their possible use as anti-caries agents.     

Influences of dissolved organic carbon on stream water chemistry in two forested catchments in central Sweden

Stream water chemistry in two headwater streams draining two small, adjacent catchments in Bispgården, central Sweden was studied during the 2003 and 2004 growing seasons. The two catchments closely resemble each other in regard to size, shape and drainage density, with the major difference found in the area of wetland lining the streambeds. The emphasis of the study was to investigate the stream water chemistry of these closely resembling catchments, regarding the quality and quantity of dissolved organic carbon (DOC) and its influence on the concentrations of di- and trivalent cations. The streams showed significant differences in the content and size distribution of DOC and in the distribution of cations between the different size fractions. For both streams the high flow events induced by precipitation influenced the chemistry of the streams through increase of organic matter and its associated cations. Fanbergsbäcken, with relatively low pH and high DOC concentration, had a greater amount of high molecular mass (HMM) DOC to which approximately 75% of Al and Fe and about 50% of Ca and Mg were associated. Gråbergsbäcken, with a higher pH and lower DOC level, had approximately 65% of Al, 40% of Fe and 30% of Ca and Mg associated to its HMM DOC fraction. Sixteen different low molecular mass organic acids were found in the stream water, of which oxalic and lactic acid were present in the highest concentrations.     

Bacterioplankton Growth on Fractions of Dissolved Organic Carbon of Different Molecular Weights from Humic and Clear Waters

The ability of fractions of dissolved organic carbon (DOC) of different molecular weights (MW) to support bacterial growth was studied in batch culture experiments. Natural pelagic bacteria were inoculated into particle-free (0.2-μm filtered) water, taken from 10 oligotrophic lakes of differing humic content, and either used without further treatments or ultrafiltered to remove DOC of >10,000 MW or >1,000 MW. Stationaryphase abundance of bacteria in the cultures was used as an estimate of bacterial carrying capacity. High-MW DOC (>10,000) comprised an increasing fraction of total DOC with increasing total DOC and humic content of the lakes. High-MW DOC was generally more available to bacteria (i.e., more bacteria were produced per unit of organic carbon initially present) than low-MW (<10,000) DOC. The availability to bacteria of this high-MW DOC decreased with increasing humic content. However, although less available in humic lakes than in clearwater lakes, the higher abundance of high-MW DOC made it quantitatively more important as a bacterial substrate; i.e., a larger fraction of the total bacterial yield of the cultures was due to high-MW DOC compounds in humic lakes than in clearwater lakes. On the average, 48% of bacterial growth occurred at the expense of DOC of <10,000 MW. DOC of <1,000 MW was responsible for an average of 22% of bacterial growth, with no significant correlation to humic content and DOC concentration of lakes. The DOC which supports bacterial growth, as well as the total DOC, is of different quality in humic and clearwater lakes.     

Effects of the Dry–Wet Hydrological Shift on Dissolved Organic Carbon Dynamics and Fate Across Stream–Riparian Interface in a Mediterranean Catchment

Abstract The stream–riparian interface, characterized by a dynamic and complex hydrology, is an important control point for nutrient fluxes and processing between terrestrial and aquatic systems. Predicted alterations in the discharge regime in Mediterranean climate regions make it necessary to understand the effects of abrupt hydrological transition between dry and wet conditions on the transport and fate of dissolved organic carbon (DOC) across the stream–riparian interface. In this study, the concentrations and fate of total DOC (TDOC) and a subset of four molecular weight fractions (<1 kDa, 1–10 kDa, 10–100 kDa, >100 kDa) were investigated in stream water and riparian groundwater during autumn of 2003 and 2004. The two study periods were characterized by contrasting antecedent hydrological conditions: the streamflow was interrupted in summer 2003 but was permanent in summer 2004. Comparison of the two study periods indicates that an abrupt dry–wet hydrological transition amplifies the water exchange across the stream–riparian interface and favors retention of up to 57% of the TDOC that flows across the interface. Furthermore, the efficiency of DOC retention across the stream-riparian interface also varies greatly depending on DOC molecular size. More than 70% of DOC fractions higher than 10 kDa were retained, whereas the smaller fraction (less than 1 kDa) was nearly conserved. Consequently, our study helps to clarify the effects of extreme hydrological events on DOC transport in running waters in Mediterranean regions.     

Longitudinal patterns of dissolved organic carbon concentration and suspended bacterial density along a blackwater river

Dissolved organic carbon (DOC) is the dominant form of carbon in transport in blackwater rivers, and bacteria are the major biological agents of its utilization. This study describes longitudinal patterns in DOC concentration and relates them to suspended bacterial populations in the channel. Concentrations of total DOC, three molecular weight fractions, and bacterial numbers were determined at 12 sites along the Ogeechee River in 1985–1986 and 1989 during periods of low and high discharge. Suspended bacterial populations were compared with DOC concentrations to determine if differences in bacterial abundance were related to longitudinal patterns of DOC concentration. Three distinct longitudinal patterns were observed: (1) The longitudinal pattern followed by both total and intermediate molecular weight DOC concentrations was a linear function of the geographic distance along the river. (2) During low flow conditions, there was a high degree of correspondence between patterns of bacterial numbers and low MW DOC (< 1000 apparent MW). (3) During periods of high discharge, the proportion of high (> 10,000) and intermediate (1000–10,000) MW fractions increased, and there was no longer a clear relationship between bacterial cells and low MW DOC.     

Fluorescence and amino acid characteristics of molecular size fractions of DOM in the waters of Lake Biwa

Dissolved organic matter (DOM) in the waters from Lake Biwa, Japan was fractionated using tangential flow ultrafiltration, and subsequently characterized by fluorescence properties and amino acids. While major dissolved organic carbon (DOC), UV absorbance (Abs), humic-like fluorescence (Flu) and total hydrolyzed amino acids (THAA) occurred in the less than 5 kDa molecular size fraction, they were not evenly distributed among various molecular size fractions. Flu/Abs ratios increased, and THAA/DOC ratios decreased with decreasing molecular size. Humic-like fluorescence occurred in all molecular size fractions, but protein-like fluorescence only occurred in the 0.1 m-GF/F fraction. Subtle differences in amino acid compositions (both individuals and functional groups) were observed between various molecular size fractions, this may indicate the occurrence of DOM degradation from higher to lower molecular weight. The results reported here have significance for further understanding the sources and nature of DOM in aquatic environments.     

Stabilization and improvement of catalytic activity of a low molar mass cellobiase by cellobiase-sucrase aggregation in the culture filtrate of Termitomyces clypeatus

The extracellular cellobiase (EC 3.2.1.21) of Termitomyces clypeatus separated in two protein fractions when culture filtrate or ammonium sulfate precipitated proteins were chromatographed on BioGel P-200 column. During purification of cellobiase (CBS) from the lower molar mass (LMM) protein fraction, the enzyme behaved like a low molecular weight multimeric protein. The purified enzyme gave a single 56 kDa band in SDS-PAGE but ladderlike bands (14, 28, 42, and 56 kDa) on denaturation by reducing-SDS and urea. The protein, however, dissociated on dilution and protomeric (14 kDa) and multimeric forms (28 and 60 kDa) were eluted separately during HPGPLC. Specific activity of CBS gradually decreased as the molar mass of the enzyme was lowered in different eluted peaks. Protein present in all CBS pool fractions had the same amino acid composition and all displayed the same, single protein peak in reverse-phase HPLC and 56 kDa band in SDS-PAGE. Thus, T. clypeatus CBS was a multimeric 14 kDa protein that was optimally active as a tetramer. CBS purified from the higher molar mass fraction (HMM) as a SDS-PAGE homogeneous 110-kDa protein did not dissociate on dilution or by SDS-urea. The purified protein was a protein aggregate as CBS consistently contained 20 +/- 5% sucrase (SUC) Units in the preparation. The aggregate resolved during reverse-phase chromatography on a C(4) column, and an additional protein peak other than CBS was detected. The aggregated CBS had a higher temperature optimum and was more stable toward thermal and chemical denaturations than SUC-free CBS. Increase of stability and catalytic activity of CBS by aggregation with SUC was much higher than those by the multimerization of CBS itself. All of these observations for the first time suggested that the heterologous protein-protein aggregation, observed for a long time for fungal enzymes, might have a significant role in modulating physicochemical properties of the extracellular enzyme.     

Lability of organic carbon in lakes of different trophic status

1. We used first‐order kinetic parameters of biological oxygen demand (BOD), the constant of aerobic decomposition (k) and the asymptotic value of BOD (BOD_ult), to characterise the lability of organic carbon pools in six lakes of different trophic state: L. Naroch, L. Miastro and L. Batorino (Belarus), L. Kinneret (Israel), L. Ladoga (Russia) and L. Mendota (U.S.A.). The relative contributions of labile and refractory organic carbon fractions to the pool of total organic carbon (TOC) in these lakes were quantified. We also determined the amounts of labile organic carbon within the dissolved and particulate TOC pools in the three Belarus lakes. 2. Mean annual chlorophyll concentrations (used as a proxy for lake trophic state) ranged from 2.3 to 50.6 μg L⁻¹, labile organic carbon (OC_L = 0.3BOD_ult) from 0.75 to 2.95 mg C L⁻¹ and k from 0.044 to 0.14 day⁻¹. 3. Our data showed that there were greater concentrations of OC_L but lower k values in more productive lakes. 4. In all cases, the DOC fraction dominated the TOC pool. OC_L was a minor component of the TOC pool averaging about 20%, irrespective of lake trophic state. 5. In all the lakes, most (c. 85%) of the DOC pool was refractory, corresponding with published data based on measurements of bacterial production and DOC depletion. In contrast, a larger fraction (27–55%) of the particulate organic carbon (POC) pool was labile. The relative amount of POC in the TOC pool tended to increase with increasing lake productivity. 6. Long‐term BOD incubations can be valuable in quantifying the rates of breakdown of the combined particulate and dissolved organic carbon pools and in characterising the relative proportions of the labile and recalcitrant fractions of these pools. If verified from a larger number of lakes our results could have important general implications.     

Can phosphorus limitation inhibit dissolved organic carbon consumption in aquatic microbial food webs? A study of three food web structures in microcosms

Microcosms with three different food web structures and phosphorus (P) limited growth medium were used to study the interactions between P and organic carbon (C) fractions in pelagic food webs. The cultures were run with low dilution to allow the biological processes to determine the outcome. A double isotope technique was used to follow the C and P compartments. In all systems the primary production was P limited. The measured P:C ratios and the observed accumulation of degradable dissolved organic carbon (DOC) indicated that the growth of heterotrophic bacteria was also P limited. The presence of neither algal grazers nor flagellates feeding on bacteria altered the limitation pattern. A net loss of P from the bacterial fraction was observed after the bloom. Different strategies for nutrient aquisition and growth are proposed as mechanisms enabling simultaneous P limitation of algae and bacteria, and a concomitant accumulation of degradable DOC. The ability of the algae to grow with low P:C ratio keeps the regeneration of P through grazers low enough to cause sustained P limitation of both algae and bacteria. The grazers were important producers of DOC when present. This implies that the usual assumption of carbon limited bacterial growth may lead to wrong conclusions regarding the dynamics of plankton communities and the DOC pool.     

Microbial control of dissolved organic carbon in lakes: research for the future

The dissolved organic carbon (DOC) of lakes dominates any budget of organic carbon in these systems. Limnologists are still limited by techniques and particularly by the lack of measurements of rates of microbial transformation and use of this DOC. There are now four different approaches to the study of the microbial control of DOC in lakes. The first is through measurements of the total DOC. Recent advances in measurement with high temperature combustion will likely lead to higher and more consistent measurements in freshwaters than previously. It is possible that a biologically active fraction may be identified. The second approach is through measurements of microbial incorporation and respiration of ¹⁴C-labeled organic matter. The kinetics of this process are well known but advances in measurement of the size of the substrate pool are still being made. The third approach is to use bacterial growth in batch or continuous flow experiments in order to understand how much of the total DOC can be decomposed by microbes. The assay in this approach may be microbial growth (thymidine incorporation, biomass changes) or change in the DOC (total concentrations, specific compounds, or fractions of the DOC by molecular weight). These methods are promising but are not developed enough for routine use. For example, growth measurements in the laboratory are all subject to experimental artifacts caused by changes in the DOC and in the bacterial populations. Finally, the fourth approach is through the use of isotopes of the natural DOC. In the sea this approach has given the age of the bulk DOC (¹⁴C data). In freshwaters it has great potential for differentiating between bacterial use of terrestrial DOC vs. use of algal-derived DOC (13C data). Stable isotopes are also useful for experimentally labeling DOC produced by algae and following the use of this material by bacteria.     

Seasonal and spatial distribution of dissolved and particulate organic carbon and bacteria in the bank of an impounding reservoir on the Enns River, Austria

1. Interstitial bacterial abundance, production and ectoenzyme activity were investigated over an annual cycle in an Austrian river when infiltration of oligotrophic river water into a river-bank was artificially enhanced. These microbial parameters were related to porewater chemistry and the concentration of particulate (POC) and dissolved organic carbon (DOC).
2. Porewater chemistry reflected the hydrodynamic mixing of infiltrating river water with riparian groundwater. Seasonal fluctuations in the microbial parameters resulted mainly from changes in temperature and organic matter supply. Seasonal change in porewater chemistry in the river-bank was detectable laterally only within the first metre of the sediment and decreased rapidly with increasing distance from the sediment–water interface.
3. The DOC concentration decreased only slightly during lateral transport through the aquifer, while total organic carbon (TOC) concentration as well as abundance and activity of interstitial bacteria were reduced by up to one order of magnitude within the top metre of the sediment. Retention of incoming particulate matter structured the lateral distribution pattern of TOC concentration. The POC and not the DOC pool was the main source of carbon for interstitial bacteria and, therefore, the quality of POC determines the distribution of microbial metabolism within the riparian zone.     

Nutrient transfer between parafluvial interstitial water and river water: influence of gravel bar heterogeneity

1. Nutrient concentrations in interstitial water springs at the downstream ends of two gravel bars in the River Rhône were measured in order to estimate the importance of nutrient inputs [nitrate, dissolved organic carbon (DOC), biodegradable and refractory fractions of DOC] from the parafluvial zone (saturated sediments adjacent to the wetted channel, i.e. interstitial habitats of gravel bars) to the river as well as seasonal variations in these inputs.
2. Compared with surface water, parafluvial water had lower concentrations of DOC (especially the refractory fraction) and generally higher concentrations of nitrate. These differences were at their lowest during winter.
3. The influence of gravel bar geomorphological heterogeneity (grain size and spatial distribution of fine sediments) on parafluvial inputs was studied in terms of nutrient content and biofilm characteristics along transects across the gravel bars.
4. A gravel bar located in a stable area of the river had low transversal heterogeneity and acted as a sink for DOC and as a source of nitrate. The low fine sediment content and the high oxygenation of interstitial water must have enabled aerobic processes such as mineralization of organic matter and nitrification to occur.
5. In contrast, gravel bars located in a degradation zone of the river had a spatially variable structure, acting as a sink for DOC and nitrate because localized accumulation of fine sediment and ensuing hypoxic conditions induced anaerobic processes such as denitrification.
6. This study highlights the important influence of geomorphological heterogeneity in gravel bars for nutrient transformations as well as nutrient exchanges between parafluvial interstitial systems and the adjacent river over a seasonal cycle.     

TRANSFORMATION AND ALLELOPATHY OF NATURAL DISSOLVED ORGANIC CARBON AND TANNIC ACID ARE AFFECTED BY SOLAR RADIATION AND BACTERIA1

The aim of this study was to test whether abiotic and biotic factors may affect allelopathic properties. Therefore, we investigated how solar radiation and bacteria influence allelopathic effects of the plant‐derived, polyphenolic tannic acid (TA) on microalgae. Using a block design, lake water samples with and without TA were exposed to solar radiation or kept in darkness with or without bacteria for 3 weeks. Dissolved organic carbon (DOC), specific size fractions of DOC analyzed by chromatography–organic carbon detection (LC‐OCD), and concentrations of total phenolic compounds (TPC) were measured to follow the fate of TA in lake water with natural DOC exposed to photolytic and microbial degradation. DOC and TPC decreased in dark‐incubated lake water with TA and bacteria indicating microbial degradation. In contrast, exposure to solar radiation of lake water with TA and bacteria did not decrease DOC. Chromatographic analyses documented an accumulation of DOC mean size fraction designated as humic substances (HS) in sunlit water samples with TA. The recalcitrance of the humic fraction indicates that photolytic degradation may contribute to a DOC less available for bacterial degradation. Subsequent growth tests with Desmodesmus armatus (Chodat) E. Hegewald showed low but reproducible difference in algal growth with lower algal growth rate cultured in photolytically and microbially degraded TA in lake water than cultured in respective dark treatments. This finding highlights the importance of photolytic processes and microbial degradation influencing allelopathic effects and may explain the high potential of allelochemicals for structuring the phytoplankton community composition in naturally illuminated surface waters.     

Microbial Availability and Size Fractionation of Dissolved Organic Carbon After Drought in an Intermittent Stream: Biogeochemical Link Across the Stream–Riparian Interface

The evolution of dissolved organic carbon (DOC) molecular-weight fractions, DOC biodegradability (BDOC), DOC origin [fluorescence index (FI)], and enzyme activities between the stream waters (main and ephemeral channel) and ground waters (riparian and hillslope) were analyzed during the transition from drought to precipitation in a forested Mediterranean stream. After the first rains, DOC content in stream water reached its maximum value (10–18 mg L⁻¹), being explained by the leaching of deciduous leaves accumulated on the stream bed during drought. During this period, the largest molecules (>10 kDa), were the most biodegradable, as indicated by high BDOC values measured during storm events and high enzymatic activities (especially for leucine-aminopeptidase). DOC >100 kDa was strongly immobilized (78%) at the stream–riparian interface, whereas the smallest molecules (<1 kDa) were highly mobile and accumulated in ground waters, indicating their greater recalcitrance. Differential enzymatic patterns between compartments showed a fast utilization of polysaccharides in the flowing water but a major protein utilization in the ground water. The results of the FI indicated a more terrestrial origin of the larger molecules in the flowing water, also suggesting that transformation of material occurs through the stream–riparian interface. Microbial immobilization and fast utilization of the most biodegradable fraction at the stream–riparian interface is suggested as a relevant DOC retention mechanism just after initial recharging of the ground water compartment. Large and rapid DOC inputs entering the intermittent river system during the transition from drought to precipitation provide available N and C sources for the heterotrophs. Heterotrophs efficiently utilize these resources that were in limited supply during the period of drought. Such changes in C cycling may highlight possible changes in organic matter dynamics under the prediction of extended drying periods in aquatic ecosystems.     

不同肥力棕壤溶解性有机碳、氮生物降解特性

溶解性有机碳、氮在土壤全碳、全氮含量中所占的比例很小,但却是土壤有机质中最为重要和活跃的部分。研究利用土壤溶解性有机碳、氮生物降解的测定方法,分别选取沈阳农业大学试验站不同肥力及与定位试验地紧密相连的自然林地棕壤为研究对象,开展棕壤溶解性有机碳、氮的生物降解特性的研究,为了解溶解性有机碳、氮在土壤生态系统碳、氮循环中的作用,探讨棕壤溶解性有机碳、氮与土壤肥力的关系提供理论依据。研究结果表明,棕壤林地溶解性有机碳、氮的含量最高,高肥处理次之,低肥处理含量最低。棕壤溶解性有机碳、氮与全碳、全氮和微生物量碳、氮的相关性达到极显著水平,与土壤肥力紧密相关,可以作为指示土壤肥力的重要指标。不同肥力棕壤溶解性有机碳、氮的降解速率在培养初期较快,而后逐渐减慢,降解数据符合双指数衰变模型。棕壤溶解性有机碳分别由降解速率不同的两个库组成：周转时间在1d的易分解部分和周转时间大约为400d的难分解部分。棕壤溶解性有机氮是由周转速率大约为2d的易降解部分和周转速率在99～105d左右的难分解部分组成． 经过42d的培养,浸提液中剩余溶解性有机质碳氮比值较培养前有所增加。     

Bacterial Growth in Mixed Cultures on Dissolved Organic Carbon from Humic and Clear Waters

Interactions between bacterial assemblages and dissolved organic carbon (DOC) from different sources were investigated. Mixed batch cultures were set up with water from a humic and a clear-water lake by a 1:20 dilution of the bacterial assemblage (1.0 μm of prefiltered lake water) with natural medium (sterile filtered lake water) in all four possible combinations of the two waters and their bacterial assemblages. Bacterial numbers and biomass, DOC, thymidine incorporation, ATP, and uptake of glucose and phenol were followed in these cultures. Growth curves and exponential growth rates were similar in all cultures, regardless of inoculum or medium. However, bacterial biomass produced was double in cultures based on water from the humic lake. The fraction of DOC consumed by heterotrophic bacteria during growth was in the same range, 15 to 22% of the total DOC pool, in all cultures. Bacterial growth efficiency, calculated from bacterial biomass produced and DOC consumed, was in the order of 20%. Glucose uptake reached a peak during exponential growth in all cultures. Phenol uptake was insignificant in the cultures based on the clear-water medium, but occurred in humic medium cultures after exponential growth. The similarity in the carbon budgets of all cultures indicated that the source of the bacterial assemblage did not have a significant effect on the overall carbon flux. However, fluxes of specific organic compounds differed, as reflected by glucose and phenol uptake, depending on the nature of the DOC and the bacterial assemblage.     

Terrestrial export of highly bioavailable carbon from small boreal catchments in spring floods

1. We assessed the terrestrial export of organic carbon, which effectively supported aquatic bacterial production (BP), from small boreal catchments during spring flood. We analysed stream runoff from nine small catchments with different proportions of peat mires and coniferous forests by monitoring the dissolved organic carbon (DOC) flux in combination with conducting bacterial bioassays.
2. Multiple linear regression analysis showed that BP during 7-day-dark bioassays (BP₇; μg C L⁻¹day⁻¹) was explained by both the quantity and quality (low-molecular weight fractions) of the DOC. BP₇ can be used as a measure of export of terrestrial organic carbon that is highly bioavailable.
3. Total export of DOC during spring flood from the different catchments ranged from 20 to 27 kg ha⁻¹ and was negatively correlated to forest cover (%). However, the export of BP₇ carbon was positively correlated to forest cover and varied from about 0.1 kg ha⁻¹ in mire-dominated streams to about 0.2 kg ha⁻¹ in forest-dominated streams.
4. The high bioavailability of forest carbon suggests that forests are the main contributors of BP-supporting carbon in boreal streams although mires have higher area-specific export of DOC.     

Identifying sources of dissolved organic carbon in agriculturally dominated rivers using radiocarbon age dating: Sacramento–San Joaquin River Basin, California

We used radiocarbon measurements of dissolved organic carbon (DOC) to resolve sources of riverine carbon within agriculturally dominated landscapes in California. During 2003 and 2004, average Δ¹⁴C for DOC was ?254‰ in agricultural drains in the Sacramento–San Joaquin Delta, ?218‰ in the San Joaquin River, ?175‰ in the California State Water Project and ?152‰ in the Sacramento River. The age of bulk DOC transiting the rivers of California’s Central Valley is the oldest reported for large rivers and suggests wide-spread loss of soil organic matter caused by agriculture and urbanization. Using DAX 8 adsorbent, we isolated and measured ¹⁴C concentrations in hydrophobic acid fractions (HPOA); river samples showed evidence of bomb-pulse carbon with average Δ¹⁴C of 91 and 76‰ for the San Joaquin and Sacramento Rivers, respectively, with older HPOA, ?204‰, observed in agricultural drains. An operationally defined non-HPOA fraction of DOC was observed in the San Joaquin River with seasonally computed Δ¹⁴C values of between ?275 and ?687‰; the source of this aged material was hypothesized to be physically protected organic-matter in high clay-content soils and agrochemicals (i.e., radiocarbon-dead material) applied to farmlands. Mixing models suggest that the Sacramento River contributes about 50% of the DOC load in the California State Water Project, and agricultural drains contribute approximately one-third of the load. In contrast to studies showing stabilization of soil carbon pools within one or two decades following land conversion, sustained loss of soil organic matter, occurring many decades after the initial agricultural-land conversion, was observed in California’s Central Valley.     

Nanostructures of APOBEC3G support a hierarchical assembly model of high molecular mass ribonucleoprotein particles from dimeric subunits

Human APOBEC3G (hA3G) is a cytidine deaminase that restricts human immunodeficiency virus (HIV)-1 infection in a vif (the virion infectivity factor from HIV)-dependent manner. hA3G from HIV-permissive activated CD4+ T-cells exists as an inactive, high molecular mass (HMM) complex that can be transformed in vitro into an active, low molecular mass (LMM) variant comparable with that of HIV-non-permissive CD4+ T-cells. Here we present low resolution structures of hA3G in HMM and LMM forms determined by small angle x-ray scattering and advanced shape reconstruction methods. The results show that LMM particles have an extended shape, dissimilar to known cytidine deaminases, featuring novel tail-to-tail dimerization. Shape analysis of LMM and HMM structures revealed how symmetric association of dimers could lead to minimal HMM variants. These observations imply that the disruption of cellular HMM particles may require regulation of protein-RNA, as well as protein-protein interactions, which has implications for therapeutic development.