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Sex reversal caused by Mus musculus domesticus Y chromosomes linked to variant expression of the testis-determining gene Sry.

When the Y chromosomes from certain populations of Mus musculus domesticus are introduced into the mouse strain C57BL/6 (B6), testis determination can fail, resulting in gonads developing either as ovotestes (with both ovarian and testicular components) or as ovaries. Not all Y(DOM) chromosomes cause sex reversal. Y(DOM) chromosomes are divided into three classes based upon their ability to induce testes in B6. The molecular basis underlying the three Y(DOM) classes is an enigma. The simplest explanation is that they harbor different alleles of the testis-determining gene, Sry. Sequencing of Sry(DOM) genes has indeed identified polymorphisms. However, none were unequivocally linked to the sex-reversal trait. It was concluded that all SRY(DOM) proteins are functionally equivalent. Using a semiquantitative RT-PCR assay, we now show that representatives of the three Y(DOM) classes have variant Sry expression patterns, that severity of sex reversal correlates with Sry mRNA titers, and that genetic correction of the sex reversal results in the upregulation of Sry expression. We propose that the variant Sry expression patterns result from polymorphisms at the site of a putative Sry enhancer.

3' Untranslated Regions↗

Human homology and candidate genes for the Dominant megacolon locus, a mouse model of Hirschsprung disease.

Hirschsprung disease (HSCR) is a congenital disorder of the enteric nervous system characterized by the absence of enteric ganglia. Three genes for HSCR have been identified: the RET proto-oncogene, the gene coding for the endothelin B receptor (EDNRB), and the endothelin 3 gene (EDN3). In mice, natural and in vitro-induced mutations affecting the Ret, Ednrb, and Edn3 genes generate a phenotype similar to human HSCR. Another model of HSCR disease is the Dominant megacolon (Dom), a spontaneous mouse mutation for which the target gene has not yet been identified. The Dom mutation has been mapped to the middle-terminal region of mouse chromosome 15, between D15Mit68 and D15Mit2. Using new or known polymorphisms for conserved human/mouse genes, we established the homology between the Dom locus and human chromosome 22q12-q13. Two genes, Smstr3 and Adsl, not previously mapped in the mouse genome, were located on mouse Chromosome 15. Three genes (Smstr3, Lgals1, and Pdgfb) are possible Dom candidates, as they do not recombine with the Dom mutation in a 252 Dom/+ animal backcross.

Animals↗

Domoic acid inhibits adenylate cyclase activity in rat brain membranes.

Adenylate cyclase activity measured by the formation of cyclic AMP in rat brain membranes was inhibited by a shellfish toxin, domoic acid (DOM). The inhibition of enzyme was dependent on DOM concentration, but about 50% of enzyme activity was resistant to DOM-induced inhibition. Rat brain supernatant resulting from 105,000 x g centrifugation for 60 min, stimulated adenylate cyclase activity in membranes. Domoic acid abolished the supernatant-stimulated adenylate cyclase activity. The brain supernatant contains factors which modulate adenylate cyclase activity in membranes. The stimulatory factors include calcium, calmodulin, and GTP. In view of these findings, we examined the role of calcium and calmodulin in DOM-induced inhibition of adenylate cyclase in brain membranes. Calcium stimulated adenylate cyclase activity in membranes, and further addition of calmodulin potentiated calcium-stimulated enzyme activity in a concentration dependent manner. Calmodulin also stimulated adenylate cyclase activity, but further addition of calcium did not potentiate calmodulin-stimulated enzyme activity. These results show that the rat brain membranes contain endogenous calcium and calmodulin which stimulate adenylate cyclase activity. However, calmodulin appears to be present in membranes in sub-optimal concentration for adenylate cyclase activation, whereas calcium is present at saturating concentration. Adenylate cyclase activity diminished as DOM concentration was increased, reaching a nadir at about 1 mM. Addition of calcium restored DOM-inhibited adenylate cyclase activity to the control level. Similarly, EGTA also inhibited adenylate cyclase activity in brain membranes in a concentration dependent manner, and addition of calcium restored EGTA-inhibited enzyme activity to above control level.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclase Inhibitors↗

Enhanced temporal summation of pressure pain in the trapezius muscle after delayed onset muscle soreness.

Temporal summation of muscle pain is an important factor in musculoskeletal pain as central integration of repetitive nociceptive input can be facilitated in musculoskeletal pain patients. The aim of this study is to evaluate changes in temporal summation of pressure pain after induction of delayed onset muscle soreness (DOMS) of the trapezius muscle. Sixteen healthy volunteers participated in the study. Temporal summation of pain was induced by sequential pressure stimulation by a computer-controlled algometer. Sequential stimulation consisting of ten stimuli (at pressure pain threshold intensity) was applied over the trapezius muscle. Stimulus duration was 1 s and inter-stimulus intervals (ISI) were 1, 5, 10, and 30 s, respectively. The pain was rated on a continuous visual analogue scale (VAS, 10 cm) after each stimulus and normalised to the VAS score from the first stimulus. DOMS was induced in the right trapezius muscle by eccentric shoulder exercises while the left trapezius muscle served as control. Temporal summation of pressure evoked pain was measured before and 24 h after the exercise. At 24 h after exercise, soreness intensity during shoulder elevation was 3.7+/-0.2 cm, while no soreness was observed on the control side. When sequential pressure stimulation was applied to the DOMS muscle, VAS scores for 1 s ISI progressively increased to a higher level than before exercise (VAS increase for the last stimulus: 0.8+/-0.2 cm vs. 0.6+/-0.1 cm, P<0.05), while VAS scores for ISI 5, 10, and 30 s were not increased. On the control side, significant increases in VAS scores was observed for all ISIs but not affected by contralateral DOMS. Facilitation of temporal summation for 1 s ISI indicated that DOMS may increase the central excitability besides involving peripheral sensitisation. During DOMS there was no potential for further nociceptor sensitisation by repeated noxious pressure stimuli, which may account for the diminishment of temporal summation evoked by pressure stimuli with ISI 5, 10, and 30 s. These data indicate that muscle soreness might facilitate the central components of temporal summation to mechanical stimulation.

Adult↗

Bacterial Growth on Photochemically Transformed Leachates from Aquatic and Terrestrial Primary Producers.

We measured bacterial growth on phototransformed dissolved organic matter (DOM) leached from eight different primary producers. Leachates (10 mg C liter(-1)) were exposed to artificial UVA + UVB radiation, or kept in darkness, for 20 h. DOM solutions were subsequently inoculated with lake water bacteria. Photoproduction of dissolved inorganic carbon (DIC), ranging from 3 to 16 µg C liter(-1) h(-1), and changes in the absorptive characteristics of the DOM were observed for all leachates upon UV irradiation. The effects of irradiation exposure on DOM bioavailability varied greatly, depending on leachate and type of bacterial growth criterion. Bacterial carbon utilization (biomass production plus respiration) over the entire incubation period (120 h) was enhanced by UV radiation of leachate from the terrestrial leaves, relative to carbon utilization in non-irradiated leachates. Conversely, carbon utilization was reduced by radiation of the leachates from aquatic macrophytes. In a separate experiment, the stable C and N isotope composition of bacteria grown on irradiated and non-irradiated DOM was estimated. Bacterial growth on UV-irradiated DOM was enriched in (13)C relative to the bacteria in the non-irradiated treatments; this result may be explained by selective assimilation of photochemically produced, isotopically enriched labile compounds.

Journal Article↗

Further glycogen decrease during early recovery after eccentric exercise despite a high carbohydrate intake.

BACKGROUND: Delayed onset muscle soreness (DOMS) is a well-known phenomenon of athletes. It has been reported from muscle biopsies that the rate of muscle glycogen resynthesis is reduced after eccentric compared to concentric exercise. AIM OF THE STUDY: Try to compensate by a carbohydrate (CHO)-rich diet the decelerated glycogen resynthesis after eccentric exercise, measured by magnetic resonance spectroscopy. METHODS: Glycogen, phosphocreatine, ATP, and Pi were measured in the human calf muscle. Twenty athletes divided into two groups (DOMS and CONTROL), reduced glycogen in M. gastrocnemius during two different running protocols. Additionally, 12 DOMS subjects performed an eccentric exercise while the CONTROL group rested. Subsequently, subjects consumed a CHO-rich diet (> 10 g/kg body mass/24 h). RESULTS: In both groups, glycogen has been reduced by about 50%. The first 2 h after exercise, glycogen dropped further (-15.6 +/- 15.7 mmol/ kg ww) in the DOMS but rose by +18.4 +/- 20.8 mmol/kg ww in the CONTROL group (P < 0.001). CONTROL subjects reached resting glycogen within 24 h (137 +/- 47 mmol/kg ww), while DOMS subjects needed more than one day (91 +/- 23 mmol/kg ww; P < 0.001). Pi and Pi/PCr, indicators of muscle injury, rose significantly in the DOMS but not in the CONTROL group. CONCLUSION: The diet rich in CHO's was not able to refill glycogen stores after eccentric exercise. Glycogen decreased even further during the beginning of recovery. This loss, which to our knowledge has not been measured before is probably the consequence of muscle cell damage and their reparation.

Adult↗

Association of Chernobyl-derived 239+240Pu, 241Am, 90Sr and 137Cs with different molecular size fractions of organic matter in the soil solution of two grassland soils.

Radiocesium is normally bound only rather weakly and unspecifically by humic substances, in contrast to the actinides Pu and Am. Recently, however, it was observed that fallout 137Cs in the soil solution from an Of-horizon of a podzol forest soil (slightly decomposed plant material) was associated essentially only with one single size fraction of the humic substances. In deeper soil layers with well humified material (AOh-horizon), radiocesium was associated with all size fractions of the dissolved organic matter (DOM). To examine whether this unexpected behaviour is also observable for DOM isolated from other soils, we determined the association of fallout 137Cs, 90Sr, 238Pu, 239+240Pu and 241Am with various size fractions of DOM from in situ soil solutions isolated from two layers (0-2 cm and 2-5 cm) of two grassland soils (a soddy podzolic soil and a peat soil) within the 10 km zone of the nuclear reactor at Chernobyl (Ukraine). The four size fractions of DOM as obtained by gel filtration of the soil solution were (mean nominal molecular weight in daltons): fraction I: > or = 2000, fraction II: 1300; fraction III: 560, fraction IV: inorganic compounds. The results for the well humified DOM (humus accumulation horizon of podzol, deeper layer of peat soil) showed that Pu and Am are essentially associated with the high molecular weight fractions, while Sr is present only in the 'inorganic' fraction. Radiocesium is found in all the size fractions separated. A quite similar pattern was also found for Pu, Am, and Sr in the soil solution from only slightly decomposed plant material (0-2 cm of peat soil), but not for radiocesium. This radionuclide was again essentially only observable in one single low molecular weight fraction of DOM. The above results thus support our recent observations in the different horizons of a forest podzol mentioned above, even though no reason for the different binding of radiocesium by well humified soil organic matter and by only slightly decomposed plant material can be given at present. The data demonstrate, however, that information on only the total amount of a radionuclide in the soil solution will not be sufficient to interpret or predict its fate adequately in the soil.

Americium↗

Eccentric exercise, isokinetic muscle torque and delayed onset muscle soreness: the role of reactive oxygen species.

There is growing evidence that reactive oxygen species (ROS) are involved in the muscular damage and soreness that is observed following strenuous or unaccustomed exercise. This study investigated the relationship between delayed onset muscle soreness (DOMS), muscle function and ROS following downhill running using electron spin resonance (ESR) spectroscopy and plasma malonaldehyde (MDA) concentrations. Eight physically active male subjects participated in two trials consisting of 30 min of running at approximately 65% VO(2max) on the flat (FLA) or a 15% downhill (DWN) gradient. Venous blood samples were drawn before, immediately after, and then 24, 48 and 72 h post exercise, and at the same time DOMS and muscle function were assessed. Blood was analysed for markers of ROS, total and differential white blood cell count, and creatine kinase. Muscle function was measured on an isokinetic dynamometer, whilst DOMS was assessed using a visual analogue scale. An increase in ROS, detected via ESR spectroscopy and MDA, was observed following DWN ( P<0.05) but not following FLA. Increased DOMS and loss of muscle function were observed following DWN ( P<0.05) but not following FLA ( P>0.05). DWN resulted in a transient leukocytosis ( P<0.05) occurring immediately post-exercise but returning to pre-exercise levels by 24 h. Although DWN resulted in an increase in ROS production, the increase occurred after the peak decline in muscle function and DOMS, suggesting that there may be a disassociation in the temporal relationship between ROS, loss of muscle function and DOMS.

Adult↗

Markers of inflammation and myofibrillar proteins following eccentric exercise in humans.

The purpose of this study was to examine the time-course and relationships of technetium-99m (99mTc) neutrophils in muscle, interleukin-6 (IL-6), myosin heavy chain fragments (MHC), eccentric torque, and delayed onset muscle soreness (DOMS) following eccentric exercise in humans. Twelve male subjects completed a pre-test DOMS questionnaire, performed a strength test and had 100 ml blood withdrawn for analysis of plasma IL-6 and MHC content. The neutrophils were separated, labelled with 99mTc, and re-infused into the subjects immediately before the exercise. Following 300 eccentric repetitions of the right quadriceps muscles on an isokinetic dynamometer, the subjects had 10 ml of blood withdrawn with repeated the eccentric torque exercise tests and DOMS questionnaire at 0, 2, 4, 6, 20, 24, 48, 72 h, and 6 and 9 days. Bilateral images of the quadriceps muscles were taken at 2, 4, and 6 h. Computer analysis of regions of interest was used to determine the average count per pixel. The 99mTc neutrophils and IL-6 increased up to 6 h post-exercise (P < 0.05). The neutrophils were greater in the exercised muscle than the non-exercised muscle (P < 0.01). The DOMS was increased from 0 to 48 h, eccentric torque decreased from 2 to 24 h, and MHC peaked at 72 h post-exercise (P < 0.001). Significant relationships were found between IL-6 and 2 h and DOMS at 24 h post-exercise (r = 0.68) and assessment of the magnitude of change between IL-6 and MHC (r = 0.66). These findings suggest a relationship between damage to the contractile proteins and inflammation, and that DOMS is associated with inflammation but not with muscle damage.

Adult↗

[Delayed post effort muscle soreness].

Muscle intolerance to exercise may result from different processes. Diagnosis involves confirming first the source of pain, then potential pathological myalgia. Delayed-onset muscle soreness (DOMS), commonly referred as tiredness, occurs frequently in sport. DOMS usually develops 12-48 h after intensive and/or unusual eccentric muscle action. Symptoms usually involve the quadriceps muscle group but may also affect the hamstring and triceps surae groups. The muscles are sensitive to palpation, contraction and passive stretch. Acidosis, muscle spasm and microlesions in both connective and muscle tissues may explain the symptoms. However, inflammation appears to be the most common explanation. Interestingly, there is strong evidence that the progression of the exercise-induced muscle injury proceeds no further in the absence of inflammation. Even though unpleasant, DOMS should not be considered as an indicator of muscle damage but, rather, a sign of the regenerative process, which is well known to contribute to the increased muscle mass. DOMS can be associated with decreased proprioception and range of motion, as well as maximal force and activation. DOMS disappears 2-10 days before complete functional recovery. This painless period is ripe for additional joint injuries. Similarly, if some treatments are well known to attenuate DOMS, none has been demonstrated to accelerate either structural or functional recovery. In terms of the role of the inflammatory process, these treatments might even delay overall recovery.

Exercise↗

Muscle damage is linked to cytokine changes following a 160-km race.

Muscle damage and perceived soreness following the 160-km Western States Endurance Run were related to changes in plasma cytokines and use of nonsteroidal anti-inflammatory drugs (NSAIDS). Subjects included 60 ultramarathoners (mean+/-SE, age 45.3 +/- 1.1 years) who finished the race in under 30 h (26.3 +/- 0.4 h). Blood samples were collected the morning prior to and immediately following the race, and subjects recorded muscle soreness during the week following the race using a 10-point Likert scale (DOMS). Seven plasma cytokines were measured including IL-6, IL-10, IL-8, IL-1ra, granulocyte colony-stimulating factor (G-CSF), monocyte chemotactic protein 1 (MCP-1), and macrophage inflammatory protein 1beta (MIP-1beta). Cytokine changes were compared between NSAID users and nonusers, and correlated with creatine phosphokinase (CPK) and DOMS. Significant increases were measured for all seven cytokines, with the greatest fold increases seen for IL-6 (125x), IL-10 (24x), and G-CSF (12x). CPK was correlated with changes in IL-6, G-CSF, IL-10, IL-1ra, and MCP-1 (r = .49-.68), (P < .001), but not IL-8 or MIP-1beta. DOMS averaged 7.1 +/- 0.3 the day after the race, and 5.0 +/- 0.3, 2.5 +/- 0.2, and 1.6 +/- 0.1 3 days, 5 days, and 7 days post-race, respectively, and each was correlated with CPK (r = .40-.63, P < .001) and changes in IL-6, G-CSF, IL-10, and MCP-1 (r = .28-.77, P < .05). A comparison of NSAID users (72% of athletes) and nonusers showed no differences in CPK or DOMS, but did reveal greater increases in five of seven cytokines in the NSAID users (P < .05). In conclusion, muscle damage in athletes competing in a 160-km race was significantly correlated with post-race DOMS and increases in five of seven cytokines. NSAID users did not experience a reduction in muscle damage or DOMS, but did have higher post-race plasma levels in five of seven cytokines.

Adult↗

Reverse osmosis sampling does not affect the protective effect of dissolved organic matter on copper and zinc toxicity to freshwater organisms.

Dissolved organic matter (DOM) plays a significant role in protecting freshwater organisms against metal toxicity. To study this, reverse osmosis (RO) has been widely used as a highly efficient method for rapid collection of large quantities of DOM from natural surface waters. The objective of this study was to examine the potential impact of the RO isolation technique on the protective effects of DOM on the toxicity of copper and zinc to the cladoceran Daphnia magna and the green alga Pseudokirchneriella subcapitata. DOM was concentrated from a natural surface water using RO and at the same time a natural (unconcentrated) surface water was taken. The concentrated DOM was rediluted to the level of the natural water to obtain the so-called reconstituted water. Chemical analyses and toxicity tests were performed with both the natural surface water and the reconstituted water. First, most chemical parameters were not significantly changed by the RO sampling. For both copper and zinc, no significant differences were observed in 48 h-EC50s for D. magna and in 72 h-EC50s for P. subcapitata between the reconstituted water and the natural water. Hence, it may be concluded that reverse osmosis does not significantly affect the protective effect of natural DOM against copper and zinc toxicity.

Animals↗

Transformations of dissolved organic matter in a landfill leachate--a size exclusion chromatography/mass spectrometric approach.

The aim of the present study was to search for qualitative changes in the landfill leachate DOM along a groundwater gradient. The study was focused on DOM characteristics of importance for its interaction with pollutants, such as molecular weight distribution and aromaticity. It was concluded that the leachate DOM underwent substantial qualitative changes along the investigated gradient at the Vejen landfill, Denmark. The molecular weight decreased, the polydispersity increased, and the aromaticity varied with the lowest values found in the middle of the gradient. The high aromaticity in the end of the gradient may explain the higher DOM binding capacity towards hydrophobic compounds seen earlier in these samples. The relative abundance of ions with mass to charge ratio (m/z) of 600-1200 seemed to be very stable along the gradient, indicating that the observed qualitative changes of the DOM is mostly attributed to changes in the m/z 100-600 range. The DOM seemed to become more similar to fulvic acids present in uncontaminated groundwater with respect to molecular weight and polydispersity along the gradient.

Benzopyrans↗

Fractionation of dissolved organic matter in mature landfill leachate and its recycling by ultrafiltration and evaporation combined processes.

XAD resin procedure was used to isolate dissolved organic matter (DOM) of a mature leachate collected from a municipal solid waste landfill in Beijing, China. The fulvic acids fraction dominated in collected leachate, accounting for 64% of DOM as dissolved organic carbon (DOC), and a high concentration of humic substances (HS) with a value of 441 mg l(-1) was found in the leachate. Molecular size distribution of DOM was also done by using batch ultrafiltration (UF) technique. Two major parts of the molecules of DOM were distributed in the regions of smaller than 1 kDa and 1-3.5 kDa, containing 45% and 43% of DOM as DOC, respectively. The laboratory-scale experiments were conducted to investigate and to develop an overall concept for mature leachate recycling based on UF and evaporation combined processes. Separation factor (SF) was applied to evaluate separation efficiency of UF. The data indicate that SF is strongly correlated with concentration factor. Second retentate from two-stage UF contained relatively high amount of DOM comprising 91% of HS and an insignificant concentration of heavy metals. The SF value for two-stage UF was more than 10. These results demonstrate that two-stage UF could effectively separate and refine HS in mature leachate from inorganic components. For achieving the main requirements as a fertilizer product, second retentate was further concentrated by evaporation, and contents of organic matter and dry-solids were obtained 13 wt.% and 25 wt.%, respectively. This study confirmed the feasibility of recycling HS from mature leachate.

Membranes, Artificial↗

Sorption of phenanthrene by soils contaminated with heavy metals.

The fate of polycyclic aromatic hydrocarbons (PAHs) in soils with co-contaminants of heavy metals has yet to be elucidated. This study examined sorption of phenanthrene as a representative of PAHs by three soils contaminated with Pb, Zn or Cu. Phenanthrene sorption was clearly higher after the addition of heavy metals. The distribution coefficient (K(d)) and the organic carbon-normalized distribution coefficient (K(oc)) for phenanthrene sorption by soils spiked with Pb, Zn or Cu (0-1000 mg kg(-1)) were approximately 24% larger than those by unspiked ones, and the higher contents of heavy metals added into soils resulted in the larger K(d) and K(oc) values. The enhanced sorption of phenanthrene in the case of heavy metal-contaminated soils could be ascribed to the decreased dissolved organic matter (DOM) in solution and increased soil organic matter (SOM) as a consequence of DOM sorption onto soil solids. Concentrations of DOM in equilibrium solution for phenanthrene sorption were lower in the case of the heavy metal-spiked than unspiked soils. However, the decreased DOM in solution contributed little to the enhanced sorption of phenanthrene in the presence of metals. On the other hand, the sorbed DOM on soil solids after the addition of heavy metals in soils was found to be much more reactive and have far stronger capacity of phenanthrene uptake than the inherent SOM. The distribution coefficients of phenanthrene between water and the sorbed DOM on soil solids (K(ph/soc)) were about 2-3 magnitude larger than K(d) between water and inherent SOM, which may be the dominant mechanism of the enhanced sorption of phenanthrene by soils with the addition of heavy metals.

Adsorption↗

Photochemical alteration of the molecular weight of dissolved organic matter.

Molecular weight (MW) is a key control on the physical, chemical, and biological characteristics of dissolved organic mater (DOM). This study investigated the effect of photooxidation on the average MW of DOM by exposing DOM of diverse origins to simulated solar radiation at varying levels of dissolved oxygen and under different incident light wavelength regimes. During irradiation, high-molecular-weight fractions were destroyed and low-molecular-weight constituents were formed. The average MW decreased with irradiation time in all treatments in a manner that can be described by a quasi-exponential function, which suggests that solar radiation is incapable of completely mineralizing DOM even after prolonged exposure. Increasing the oxygen concentration accelerated the MW reduction while the removal of oxygen strongly suppressed this process. The fractional contributions from UV-B, UV-A, and visible radiations to full-spectrum photoinduced MW reduction varied considerably among the DOM samples examined, ranging from 19% to 60%, 17% to 36%, and 15% to 46%, respectively. The MW changes in time-series irradiations were inversely correlated with the ratio of the absorbance at 250 nm to that at 365 nm (i.e., the E(2)/E(3) quotient). Photoinduced MW reduction was accompanied by a decrease of polydispersity, which is indicative of a reduced DOM heterogeneity.

Hydrogen-Ion Concentration↗

Nickel speciation in the presence of different sources and fractions of dissolved organic matter.

This study evaluated nickel (Ni) speciation in the presence of different fractions (humic acid (HA), fulvic acid (FA)) and sources (natural sediment, Suwannee River, peat moss) of dissolved organic matter (DOM) at Ni concentrations toxicologically relevant to the freshwater amphipod, Hyalella azteca. The free Ni ion, Ni(2+), was measured in reconstituted water (with or without DOM) using a miniaturized ion-exchange technique (IET). Ni speciation from these experiments was compared to calculated results obtained from equilibrium modelling (WHAM, Model VI). While it is known that Ni will complex with DOM, it was found that under acutely toxic Ni exposure concentrations ([Ni(Total)]=5mg/L, or 85.1 microM) representative surface-water DOC concentrations ( approximately 10mg/L) played little or no role in Ni speciation. Conversely, at sublethal Ni exposure concentrations ([Ni(Total)]=0.2 and 0.5 microg/L, or 3.4 and 8.51 microM, respectively) DOM significantly affected Ni speciation with [Ni(2+)] decreasing with increasing concentration of DOM. It was found that for similar concentrations of DOC (same fraction, different sources), the measured Ni(2+) concentrations were reduced (relative to the control), but similar to one another. Conversely, at similar DOC concentrations, the HA fraction reduced Ni(2+) levels to a greater extent than the associated FA fraction. Overall, this study provides proof of principle that Suwannee River and peat humic substances are suitable analogues for natural sediment pore-water DOM when evaluating Ni bioavailability in freshwater.

Amphipoda↗

Discriminatory classification of natural and anthropogenic waters in two U.K. estuaries.

The ability to distinguish water inputs from both natural and anthropogenic sources was investigated in the complex environment of an urban estuary (Tyne) and a relatively pristine estuary (Tweed). We used a data set from a total of 11 estuarine transects, comprising measurements of bulk dissolved organic matter (dissolved organic carbon and nitrogen), dissolved nitrogen (total dissolved nitrogen, ammonium, nitrate+nitrite and dissolved organic nitrogen), optical absorbance measurements (a350, S290-350) and fluorescence excitation emission matrix measurements (fluorophores A, H, B and T intensity and A and H emission wavelength maxima). In order to investigate trends within the numerous parameters measured, multivariate statistics were employed. Principal components analyses showed 63.4% of the variability in the total data set can be explained by two sets of components and 74.9% of the variability by the spectrophotometric measurements alone. In both analyses the first component correlated to the mixing of terrestrial and marine waters and the second component was correlated to sources of pollution such as domestic sewage. Within the data set, river flow and terrestrially derived DOM were significantly correlated, and situations with high river input showed an increase in terrestrial signature in the estuary. Discriminant analyses were also carried out and indicated that 59.8% (total data set) and 53.3% (solely spectrophotometric data) of the samples can be correctly classified into their respective groups (water categories) assigned on the basis of salinity and sampling location. Overall the results clearly show the potential of spectrophotometric techniques to discriminate distinct water categories with different DOM characteristics. In particular, measurement of the fluorophore H emission maxima, the spectral slope parameter, S290-350, and fluorophores T and B intensity enabled discrimination of DOM from riverine, estuarine, marine, and sewage affected water categories. The results presented here indicate the ability of spectrophotometric data alone to distinguish between marine, anthropogenic and terrestrial DOM and distinguish terrestrial DOM from different catchments (Tyne vs. Tweed). With current advances in the in-situ deployment of absorbance and fluorescence spectroscopy it is anticipated that multivariate statistics will gain importance as a cost effective, powerful and diagnostic approach to assessing the distributions of water types and their associated DOM characteristics and fluxes at the land-ocean interface.

Carbon↗