Sites P4 Research Articles

A mutagenesis study of autoantigen optimization for potential T1D vaccine design

A previously reported autoreactive antigen, termed the X-idiotype, isolated from a unique cell population in Type 1 diabetes (T1D) patients, was found to stimulate their CD4+ T cells. This antigen was previously determined to bind more favorably than insulin and its mimic (insulin superagonist) to HLA-DQ8, supporting its strong role in CD4+ T cell activation. In this work, we probed HLA-X-idiotype-TCR binding and designed enhanced-reactive pHLA-TCR antigens using an in silico mutagenesis approach which we functionally validated by cell proliferation assays and flow cytometry. From a combination of single, double, and swap mutations, we identified antigen-binding sites p4 and p6 as potential mutation sites for HLA binding affinity enhancement. Site p6 is revealed to favor smaller but more hydrophobic residues than the native tyrosine, such as valine (Y6V) and isoleucine (Y6I), indicating a steric mechanism in binding affinity improvement. Meanwhile, site p4 methionine mutation to hydrophobic residues isoleucine (M4I) or leucine (M4L) modestly increases HLA binding affinity. Select p6 mutations to cysteine (Y6C) or isoleucine (Y6I) exhibit favorable TCR binding affinities, while a swap p5-p6 tyrosine-valine double mutant (V5Y_Y6V) and a p6-p7 glutamine-glutamine double mutant (Y6Q_Y7Q) exhibit enhanced HLA binding affinity but weakened TCR affinity. This work holds relevance to potential T1D antigen-based vaccine design and optimization.

Use of fatty acids as tracer of organic matter input associated with level of land urbanization.

Suspended sediments (SSs) were examined regarding the content of fatty acids (FAs) to associate them with sources of soil entry into the river. The source of organic matter was traced through fatty acid distribution, as well as erosion. Also, TOC, TN, and TOC/NT were used to support the results of FAs. For this, a tropical river was chosen to understand the main source of input considering the level of land occupation along the river. The Barigui river, in southern Brazil, was segmented in four distinct areas regarding the soil occupation (P1, P2, P3, and P4). Nine sampling campaigns were conducted from Nov/2014 to Nov/2015 using a time-integrated sampler. Site P1 has the lowest level of urbanization and showed the lowest concentration of FAs (16.35μg-1). In contrast, site P4, the most urbanized, showed the highest content of fatty acids, including those associated with erosion, 378.53μgg-1, specifically those with long chains. The mean concentrations of the saturated fatty acids (FAs) was 283.40μgg-1, monounsaturated fatty acids (MUFAs) was 79.46μgg-1, and polyunsaturated fatty acids (PUFAs) was 15.66μgg-1. Twenty-seven fatty acids were examined, nevertheless C15:0, C16:0, C18:0, and C18:1ω9 prevailed in all samples. Generally, those acids indicate sewage inputs. Statics analyses were used to find the relation between the source of organic matter (autochthonous, allochthones, and anthropogenic) and FAs. Finally, the input of organic matter is associated with land occupation, which can be distinguished by FA distribution.

Microbial risk assessment in recreational freshwaters from southern Brazil

In this study, total coliforms (TC), Escherichia coli, enterovirus (EV), rotavirus (RV), and human mastadenovirus species C and F (HAdV-C and HAdV-F) were evaluated in water samples from Belo Stream. For HAdV-C and F, the infectivity was assessed by integrated cell culture quantitative real-time polymerase chain reaction (ICC-qPCR). Samples were collected monthly (May/2015 to April/2016) at four sites. Viral analyses were performed for both ultracentrifuge-concentrated and unconcentrated samples. For site P4 (used for recreational purposes), QMRA was applied to estimate health risks associated with exposure to E. coli and HAdV-C and F. TC and E. coli were present throughout the collection period. EV and RV were not detected. HAdV-C were present in 8.51% (1.89E + 06 to 2.28E + 07 GC (Genomic Copies)/L) and 21.27% (2.36E + 05 to 1.29E + 07 GC/L) for unconcentrated and concentrated samples, respectively. For HAdV-F were 12.76% (2.77E + 07 to 3.31E + 08 GC/L) and 48.93% (1.10E + 05 to 4.50E + 08 GC/L) for unconcentrated and concentrated samples, respectively. For unconcentrated samples, infectivity for HAdV-C was detected in 37.20% (1st ICC-qPCR) and 25.58% (2nd ICC-qPCR). For HAdV-F, infectivity was detected in 6.97% (1st ICC-qPCR) and 6.97% (2nd ICC-qPCR). For concentrated samples, HAdV-C infectious was observed in 17.02% (1st ICC-qPCR) and in 8.51% (2nd ICC-qPCR). For HAdV-F, were present in 8.51% for both 1st and 2nd ICC-qPCR. Statistical analyzes showed significant difference between the collection sites when analyzed the molecular data of HAdV-F, data of TC and E. coli. Correlation tests showed direct correlation between HAdV-F with E. coli and TC. E. coli concentrations translated to the lowest estimates of infection risks (8.58E-05 to 2.17E-03). HAdV-F concentrations were associated with the highest infection risks at 9.99E-01 and for group C, 1.29E-01 to 9.99E-01. These results show that commonly used bacterial indicators for water quality may not infer health risks associated with viruses in recreational freshwaters.

Using computed tomography images to characterize the effects of soil compaction resulting from large machinery on three-dimensional pore characteristics in an opencast coal mine dump

Purpose: The use of heavy machinery in mining areas increases during mining and dumping of waste soils, causing severe soil compaction and potentially affecting soil recovery. This study aimed to capture the features of complex compacted soil pore structure using computed tomography (CT) and to quantify soil pore distribution properties based on Minkowski theory. Materials and methods: A total of 45 cylindrical soil columns were extracted from different soil layer at 5 sampling sites that differed in compaction level in Pingshuo opencast coal mine in Shanxi Province of China. The penetration resistance at these sites ranged from 27.3 to 1023.1 kPa. Three-dimensional reconstruction software VGStudio was used to obtain the morphological characteristics (pore volume density, VD; surface density, SD; mean curvature density, MCD; and Euler number, EN) of soil pores after processing the X-ray scanned image. Results and discussion: Compaction generally reduced VD, SD, and MCD, which ranged from 0.10 to 0.20, 2.23 to 2.80 mm 2 mm −3 , and 10.75 to 15.64 mm mm −3 respectively in undisturbed site P0, whereas, they ranged from 0.02 to 0.07, 0.13 to 0.60 mm 2 mm −3 , and 1.61 to 4.27 mm mm −3 respectively in highly compacted site P4. The three index exhibited the similar trend, and decreased with the increase of compaction level. Soil compaction had positive effects on EN, which was low at the depth of 20- to 40-cm layers but showed an increasing trend with increasing soil compaction level. The value of EN varied from 28,878 to 129,861 in P0, whereas it varied from 1,169,967 to 2,281,998 in P4. Conclusions: All of macroporosity, mesoporosity, and microporosity were significantly affected by heavy machinery compaction, among which macroporosity exhibited the maximum decrease in P4 compared to that in P0 (P0 ranged from 14.2 to 26.4% and P4 ranged from 0.2 to 1.4%). The soil compaction in mining area significantly affected soil properties, thus artificial methods such as fertilizer application and deep tillage were needed.

Spatial variation of metals and phosphorus in sediments of a river influenced by urbanization

ABSTRACT The suspended sediments are a special compartment that characterizes the effects of land use in the watershed. Such compartment is able to trap organic compounds as well as metals in general. Spatial variation of concentrations can be accounted for the land use, soil composition and pollution. In this work, we aimed to assess the spatial variation of metals and phosphorus concentration presented in suspended sediments collected in a river under urban occupation. Using time-integrated suspended sediment samplers, samples were collectec in three sites covering different levels of occupation: Almirante Tamandaré (P1) and two in Curitiba city (Tingui Park -P2 and Barigui Park - P3). The sampler was settled to be 20 cm below the water surface and it was removed after 30 days. Twelve samples were collected to examining the following elements Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, La, Mg, Mn, Na, Nb, Ni, P, Pb, Sc, Sn, Sr, Th, Ti, V, Y, Zn and Zr by ICP-OES. The first site is characterized with only 5.73% of urban area; meanwhile the site P3 has the highest urban area 54.06%. Among the twenty seven elements, nineteen showed a positive variation in concentration between collected sites, giving indication that an enrichment of metals along the river. The following elements showed positive variation: P (151.20%), La (117.98%), Cd (93.33%), Cr (71.43%), Na (68.75%), Ni (68.07%), Y (66.67%), Th (63.64%) and Zn (56.77%). Considering local reference values it is concluded that Cd, Cr, Ni, P, Y and Zn appear associated with urbanization increase in the basin; Co, Sc, Sn, Cu and Mn appear possibly associated with urbanization; Ba, La, Na, Pb and Sr show inconclusive behavior in relation with urban influence ; Al, Fe, Nb, Th, Ti, As, Ca, K, Mg, Zr and Va demonstrate not be associated with urbanization.

Assessment of water quality and genotoxic impact by toxic metals in Geophagus brasiliensis

This study used the pearl cichlid Geophagus brasiliensis as a bioindicator to survey the health of the aquatic environment on four sites (P1, P2, P3 and P4) of the Mumbuca stream located at Monte Carmelo/MG, Brazil. The selection of different sites was made with reference to the gradient of urban activity and via physicochemical and biological evaluation of water quality and genotoxicity. The water quality index was classified as ‘good’ for P1 and P4, regular in P2 and ‘poor’ for P3. The micronuclei (MN) frequency obtained from blood analysis was in agreement with the water quality, such that the higher values of MN were detected in sites evaluated as poor. Water degradation conditions worsen according to the flow of the stream over the sites P1, P2 and P3, but for site P4, located after the Monte Carmelo Sewage Treatment Plant, improvements in the micronuclei frequency are detected. Our results showed high levels of potentially toxic metals (chromium, lead, aluminum and nickel) in specific stream sites (P2 and P3). We suggest that the micronuclei induction in G. brasiliensis could be due to the presence of these compounds.

English

Further objectives were to determine the different patterns of land use and occupation within the microbasin formed by these rivers, and to assess the influence of the quantity of riparian vegetation on the presser/vation of the aquatic ecosystems. The studied basin extends over a total area of 77.75 km2, of which 92.98% comprised agriculture (44.11%) and native forest (48.87%). The drainage area of the sampling site P1 was 87.57% covered by crops, and so was 22.30% of the area of the sampling site P4. However, the application of the Tukey test (p < 0.05) to the data for thermotolerant coliforms (also known as faecal coliforms - FC) led to the inference of similarity (p<0.05) between the sampling sites P1 and P4. This indicates that the agriculture impact is probably mitigated by the area of native forest around the source of the spring that feeds P1. From the sampling site P2 to P3, the drainage area showed an improvement in the preservation of natural areas, with the area covered by crops decreasing to 44.41% and an increase of 47.93% in the land covered by native forest. Sampling sites P2 and P3 also showed significant similarity between their annual averages for FC, displaying a reduction in the concentration of these microorganisms from 2.15 to 1.61 (× 10³ MPN/100 mL), which again provided evidence of the positive influence of the recovery of plant cover within the drainage area. Recognising that the other sampling sites with very similar land use characteristics did not display significant alterations in the water quality, the influence of the vegetation and its role as a buffer were made evident, since for these sampling sites most water sources are not bereft of surrounding vegetation. In this way, the lack of correlation (Pearson and Spearman) between the seasonal variations for the assessed water quality parameters allowed us to provide evidence for the positive effect of riparian vegetation in the maintenance of the integrity of aquatic ecosystems and the preservation of the water quality. The water quality comprises physical, chemical, and microbiological parameters related to the different land uses within the river basin. Key words: Monitoring, total and thermotolerant coliforms, biological indicators, human impact, water quality, multifactorial analysis.

Pyrite as a proxy for the identification of former coastal lagoons in semiarid NE Brazil

This work aimed to test the suitability of pyrite (FeS2) as a proxy for reconstructing past marine environmental conditions along the semiarid coast of Brazil. Morphological description combined with physicochemical analyses including Fe partitioning were conducted for soil depth profiles (30 and 60 cm depths) at three sites in two contrasting lagoons of the state of Ceara: a suspected former lagoon that would have been transformed into a freshwater “lake” at a site vegetated by Juncus effusus (site P1), and another lagoon with connection to the sea at sites vegetated by J. effusus (site P2) or Portulaca oleracea (site P3). Soil samples were collected in September 2010. Site P3 had more reducing conditions, reaching Eh values of –132 mV in the surface layer (0–10 cm), whereas minimum values for the P1 and P2 sites were +219 and +85 mV, respectively. Lower pyritic Fe values were found at site P1, with a degree of pyritization (DOP) ranging from 10 to 13%. At sites P2 and P3, DOP ranged from 9 to 67% and from 55 to 72%, respectively. These results are consistent with an interruption of tidal channels by eolian dune migration inducing strong changes in the hydrodynamics and physicochemical characteristics (lower salinity, oxidizing conditions) of these sites, causing the dieback of suspected former mangroves and a succession to freshwater marshes with an intermediate salt marsh stage. Together with other physicochemical signatures, pyrite can evidently serve as a useful proxy in tracking environmental changes in such ecotones, with implications for coastal management.

Avaliação da toxicidade em sedimento do rio Cubatão-SP

The present paper assessed acute and chronic biological effects at sediment samples from Cubatao River and two othercontributors (Pereque and Piloes Rivers), by using ecotoxicity assays. Hyalella azteca were exposed to integral sediments.Body growing size was also determined. The samples were collected in 2010 and 2011 (four data) and nine different siteswere included, representing the Cubatao River Basin. The acute effects for Hyalella azteca were obtained at P2 and P5 (2aand 4a samplings), with mortality higher than 50% of the total number of exposed organisms. Organisms body size analysisdetected lower growing rates at sites P4 and P5, places identified as pertaining to the mainly industrialized sites.

Seasonal variability of basic pollution parameters in atmospheric precipitation in Strzelin

Abstract Electrical conductivity and pH are the basic indices of rainfall pollution, constituting the basis for evaluation of the general nature of rainwater. This article presents an analysis of these two parameters in samples of rainfall collected in the years 2008-2010 (in the heating season and outside it) in three measurement sites P1, P2 and P3, located in the city and commune Strzelin (Lower Silesian Voivodeship). Preliminary results show a high variation of pH and electrical conductivity of rainfall occurring in the analysed area. Approximately 30% of the samples were characterised by a pH that allowed to qualify them, according to JANSEN et al. [1988], as normal rainfall. A similar share in all the collected samples represented rainfall of pH &lt; 5.1 that was classified as slightly, considerably and highly acidic. The problem of acid rain occurred in the analysed area mainly during the heating season, when rainfall of pH &lt; 5.1 accounted for a considerable share in all rainfall samples collected in the said period. The electrical conductivity of most samples fell below 60 μS·cm-1, which allowed us to classify them as slightly polluted rainfall (site P2), considerably polluted (site P3) and highly polluted (site P1).

DNA Methylation Analysis of the Insulin-like Growth Factor-1 (IGF1) Gene in Swedish Men with Normal Glucose Tolerance and Type 2 Diabetes

Objective: Recent genetic studies have demonstrated that Single Nucleotide Polymorphism (SNP) rs35767(C/T) in the IGF1 gene promoter is associated with insulin resistance and serum IGF-I levels and thereby implicated that IGF1 has genetic effect in Type 2 Diabetes (T2D). The present study aimed to investigate the alteration of DNA methylation levels of the IGF1 gene in T2D. Subjects and methods: A total of 688 Swedish men with Normal Glucose Tolerance (NGT) or T2D were selected from Stockholm Diabetes Prevention Program. DNA methylation levels at rs35767 SNP-CpG site and other three CpG sites (P1-P3) in the IGF1 gene promoter region were analyzed with PyroMark Assays and bisulfite pyrosequencing. Fasting serum IGF-I levels were measured with an in-house radio-immunoassay. Results: DNA methylation levels at CpG site P3 of the IGF1 gene promoter were increased in T2D patients compared with NGT subjects (84.8% vs. 74.2%, P<0.001), while serum IGF-I levels were lower in T2D than that in NGT subjects (152 μg/l vs 169 μg/l, P=0.029). In SNP rs35767(C/T), the carriers with CC genotype had higher DNA methylation levels at SNP-CpG site compared with the carriers with CT and TT genotypes in both NGT and T2D. Conclusions: The present study provides the first evidence that increased DNA methylation levels of the IGF1 gene and decreased serum IGF-I protein concentration are associated with T2D, and suggests that DNA methylation in the IGF1 gene may interact with SNP rs35767 (C/T) in the gene promoter region.

Heavy metal pollution in soil and plants at bone char site

This study determined the heavy metal concentration in soil and plants at a bone char site in Umuahia, Nigeria. Soil and plant samples collected in a randomized complete block design (RCBD) were analyzed for zinc (Zn), lead (Pb), cadmium (Cd), nickel (Ni), and arsenic (As). The concentration of metals in soil and plants in the vicinity of the bone char site are as follows: Zn (172 mg kg−1) and Ni (0.62 mg kg−1) in soil were highest at site P3, Pb (2.37 mg kg−1) and As (0.08 mg kg−1) at site P1, and Cd (18.30 mg kg−1) at site P2. In plants, the concentrations of Zn (41.17 mg kg−1) and Cd (3 mg kg−1) were highest in Albizia ferruginea, Ni in Dialium guineense (0.09 mg kg−1), while Pb was in D. guineense (0.08 mg kg−1) and Spathodea companulata (0.06 mg kg−1). The levels of Zn, Cd, Pb, Ni, and As in soil ranged from 11.2 to 172, 2.68 to 18.2, 0.026 to 2.37, 0.33 to 0.62, and 0.02 to 0.08 mg kg−1, respectively. In plants, the concentration of Zn, Cd, Pb, and Ni ranged from 2.01 to 41.17, 0.12 to 3, 0.02 to 0.08, and 0.03 to 0.09 mg kg−1, respectively. There were significant correlations between Zn and Cd, and Pb and As in soil. The high concentration of Cd in soil might affect soil productivity.

Transcranial direct current stimulation (tDCS) produces localized and specific alterations in neurochemistry: A 1H magnetic resonance spectroscopy study

Transcranial direct current stimulation (tDCS) has been found to produce significant changes in behavior, including a large increase of learning and performance for a difficult visual perceptual task (Clark et al., NeuroImage 2010). The mechanisms by which tDCS produces these behavioral effects are currently uncertain. One hypothesis is that anodal tDCS leads to increased metabolic activity in the brain, which enhances cognitive and memory processes. Here we examined the neuronal mechanisms by which tDCS influences learning by measuring changes in brain metabolite concentrations using proton magnetic resonance spectroscopy ( 1H MRS). As perception and learning can also influence neurochemistry, here we applied tDCS during rest. MRS data was obtained before and after 2.0 mA of anodal tDCS was applied for 30 min over electrode site P4, with the cathode placed on the contralateral arm. MRS data were acquired from the right parietal lobe beneath the anodal tDCS electrode, and from the homologous regions of the left hemisphere once before and once after tDCS. Significantly higher combined glutamate and glutamine levels were found in right parietal cortex, beneath the stimulating electrode, with non-significant increases in homologous regions of the opposite hemisphere. In addition, a significant interaction between hemispheres was found for tDCS effects on tNAA. These results suggest that changes in glutamatergic activity and tNAA may be related to the mechanisms by which tDCS influences learning and behavior.

Controls on mesopelagic particle fluxes in the Sub-Antarctic and Polar Frontal Zones in the Southern Ocean south of Australia in summer—Perspectives from free-drifting sediment traps

The SAZ-Sense project examined ecosystem controls on Southern Ocean carbon export during austral summer (January–February 2007) at three locations: P1 in the low biomass Subantarctic Zone (SAZ) west of Tasmania, P3 in a region of elevated biomass in the SAZ east of Tasmania fuelled by enhanced iron supply, and P2 in High-Nutrient/Low Chlorophyll (HNLC) Polar Frontal Zone (PFZ) waters south of P1 and P3. Sinking particles were collected using (i) a cylindrical time-series (PPS3/3) trap for bulk geochemical fluxes, (ii) indented rotating sphere (IRS) traps operated as in-situ settling columns to determine the flux distribution across sinking-rate fractions, and (iii) cylindrical traps filled with polyacrylamide gels to obtain intact particles for image analysis. Particulate organic carbon (POC) flux at 150 m (PPS3/3 trap) was highest at P1, lower at P2, and lowest at P3 (3.3±1.8, 2.1±0.9, and 0.9±0.4 mmol m −2 d −1, respectively). Biogenic silica (BSi) flux was very low in the SAZ (0.2±0.2 and 0.02±0.005 mmol m −2 d −1 at P1 and P3, respectively) and much higher in the PFZ (2.3±0.5 mmol m −2 d −1 at P2). Hence, the high biomass site P3 did not exhibit a correspondingly high flux of either POC or BSi. Separation of sinking-rate fractions with the IRS traps (at 170 and 320 m depth) was only successful at the PFZ site P2, where a relatively uniform distribution of flux was observed with ∼1/3 of the POC sinking faster than 100 m d −1 and 1/3 sinking slower than 10 m d −1. Analysis of thousands of particles collected with the gel traps (at 140, 190, 240, and 290 m depth) enabled us to identify 5 different categories: fluff-aggregates (low-density porous or amorphous aggregates), faecal-aggregates (denser aggregates composed of different types of particles), cylindrical and ovoid faecal pellets, and isolated phyto-cells (chains and single cells). Faecal-aggregates dominated the flux at all sites, and were larger in size at P1 in comparison to P3. The PFZ site P2 differed strongly from both SAZ sites in having a much higher abundance of diatoms and relatively small-sized faecal-aggregates. Overall, the particle images suggest that grazing was an important influence on vertical export at all three sites, with differences in the extents of large aggregate formation and direct diatom export further influencing the differences among the sites.

P22−and P3−Units in the [Rh8P9]δ−Polyanion of La4Rh8P9

The phosphide La(4)Rh(8)P(9) was synthesized from the elements in a bismuth flux. The structure was refined from single crystal diffractometer data: space group Cmcm, a = 1303.1(2), b = 1893.2(2), c = 576.70(6) pm, wR2 = 0.0277, 1380 F(2) values, 65 variables. The rhodium and phosphorus atoms build up a three-dimensional [Rh(8)P(9)] polyanion which leaves larger cages for the three crystallographically independent lanthanum sites. The rhodium atoms have between four and six phosphorus neighbors at Rh-P distance ranging from 229 to 254 pm. Three of the four crystallographically independent phosphorus atoms are isolated (P(3-) units), while the P4 atoms form dimers with double bond character (208 pm P-P). The P(2)(2-) diphosphenide units bond side-on to a Rh3 and end-on to four Rh5 atoms. (31)P magic angle spinning (MAS) NMR spectroscopy is able to resolve three of the four crystallographically distinct phosphorus sites. The doubly bonded phosphorus site P4 is characterized by an axially symmetric shielding tensor of moderate anisotropy Δσ = σ(33) - σ(iso) = 257 ppm. Electronic band structure calculations prove the metallic character and reveal the significant difference between the isolated P(3-) and the phosphorus atoms of the P(2)(2-) units. Magnetic susceptibility measurement reveals Pauli paramagnetism.

Molecular dynamics studies of the P pilus rod subunit PapA

Adhesion of uropathogenic Escherichia coli to host tissues is mediated by pili, which extend from the outer cell membrane of the bacterium. Here we report molecular dynamics (MD) characterizations of the major constituent of P pili from the uropathogenic E. coli, PapA, in unliganded state and in complex with the G1 strand of the chaperone PapD. To mimic the PapA response to the gradual dissociation of the PapD G1 strand and to evaluate the role of PapA chaperone recognition sites, we also carried out MD simulations of complexes of PapA with fragments of PapD G1 strand, that leave either the P4 or both P3 and P4 sites unoccupied. Data on the unbound form of PapA indicate that, upon release of the chaperone, PapA evolves toward compact states that are likely not prone to subunit-subunit association. In line with recent experimental reports, this finding implies that chaperone release and subunit-subunit association must be concerted. Our data also indicated that the gradual unbinding of the chaperone from the PapA groove has increasingly strong structural consequences. Indeed, the release of the chaperone from the site P4, which is closest to the initiation site (P5), does not have dramatic effects on the domain structure, whereas its release from both the P4 and the adjacent P3 sites induces a quick structural transition toward a collapsed state, where the subunit groove is obstructed.

Complementary Approaches to Assess the Environmental Quality of Estuarine Sediments

An assessment of the environmental quality of sediments at several locations of the Ria de Pontevedra (NW of Spain) was performed by integrating toxicity data obtained from multiespecies bioassays, chemical data from analysis of mussels and sediment, and physical–chemical parameters of the sampled sites. Subsequently, a toxicity identification evaluation (TIE) method intended for characterization and identification of the toxic agents was applied to the most polluted location by using the Paracentrotus lividus sea urchin bioassay. Both metals and organic compounds seem to be the causative agents of toxicity in elutriates of the studied sediment. Finally, multivariate statistics were applied for a better interpretation of results. A factor analysis was developed to establish the relationship among variables and to derive local sediment quality guidelines (SQG) by linking chemical contamination to biological effects. When multidimensional scaling and cluster analysis were performed to group the locations according to either the chemistry or toxicity data, P3-site was always clearly broken up the others. The different approaches all supported the same conclusion: site P3 can be considered highly contaminated by both trace metals and PAHs resulting in high toxicity for all the tested species.

Mercury concentration in the sediment at different gold prospecting sites along the Carmo stream, Minas Gerais, Brazil, and frequency of resistant bacteria in the respective aquatic communities

In Brazil mercury is used indiscriminately at prospecting sites for gold extraction, with a consequent enormous discharge of this metal into the aquatic ecosystem. Mercury concentration was investigated in 1995, 1996 and 1997 in the sediments of ten sites along the Carmo stream, MG, Brazil, almost all of them located in prospecting areas. Analysis of mercury in the sediments of all sampling sites showed that the levels were above the tolerable limit (0.1μg g−1), except for sites P4 (1997), P6 (1995) and P9 (1995 and 1997). The results showed wide mercury contamination in an environment not limited to the active or inactive prospecting areas, but also including sites outside these areas but downstream from them. There was a high incidence of bacteria resistant to mercury in the aquatic communities of the sites under study, ranging from 27.3 to 77.1%, except for P1 (an ecological station upstream from the sites under study) in which all bacteria isolated from water were sensitive. Furthermore, the fall in mercury concentration in the sediment at site P5 was not as marked as at other sites, with a frequency of resistant bacteria of only 27.3%, possibly indicating a slower detoxification. The statistical analysis (Pearson’s correlation = − 0,527) showed that the hypothesis about negative correlation between the incidence of bacterial resistance and the total mercury concentration in this environment is supported. Hence, all these data denote a moderate association between the distribution of resistant bacteria and the presence of mercury compounds.

Moored observations of the current and temperature structure over the continental slope off central California: 2. The energetics of the flow off Point Sur

The data from two current meter moorings off Point Sur, California, were analyzed to examine the energy transfers over the continental slope in the California Current system (CCS). The method used was to calculate terms in the heat equation at intermediate depths between instruments, using the thermal wind relation to estimate the horizontal temperature gradients from the vertical shear. Time series of u, v, and T were collected for 17 months (May 1989 to October 1990) at site P2 on the 800‐m isobath and for 12 months (May 1990 to May 1991) at site P3, 25 km farther offshore on the 1800‐m isobath. Instrument depths were 100, 350, and 500 m at both moorings plus 1000 m at site P3. Mean values of the energy conversion terms were computed at sites P2 and P3 at 225 m depth and at site P2 at 425 m depth. The local change of eddy potential energy (EPE) was approximately balanced (∂EPE/∂t ≈ 0) for each case but the dynamics were quite different. At P2 225, the sources of EPE were advection by the mean flow and baroclinic instability, which converted mean potential energy (MPE) to EPE. The losses came from eddy advection out of the region and the vertical eddy heat flux (VEHF), which converted EPE to eddy kinetic energy (EKE). At P2 425, baroclinic instability provided the major source of EPE. This, plus a small positive contribution from the VEHF, was balanced by advection of EPE out of the region by both the mean flow and the eddies. At P3 225, mean advection and baroclinic instability were negligible, and the downward eddy heat flux (VEHF) was balanced by advection of heat out of the region by the eddies. Time series of terms in the EPE equation were used to examine high energy transfer events at both moorings. Events at P2 involved both horizontal and vertical processes and had longer timescales (several days to weeks) than those at P3, which had much shorter timescales (2–5 days) and were dominated by vertical processes. Compared to the Gulf Stream, the mean baroclinic instability at P2 (both depths) was smaller by at least a factor of 10. A term‐by‐term comparison between the mean values off Point Sur and the Kuroshio Extension showed different processes, with the eddies gaining strength at the expense of the mean flow off Point Sur and the opposite occurring in the Kuroshio Extension.

Design and study of peptide-ligand affinity chromatography adsorbents: Application to the case of trypsin purification from bovine pancreas

The purification of trypsin from bovine pancreas was employed in a case study concerning the design and optimization of peptide-ligand adsorbents for affinity chromatography. Four purpose-designed tripeptide-ligands were chemically synthesized (>95% pure), exhibiting an Arg residue as their C-terminal (site P(1)) for trypsin bio-recognition, a Pro or Ala in site P(2), and a Thr or Val in site P(3). Each tripeptide-ligand was immobilized via its N-terminal amino group on Ultrogel A6R agarose gel, which was previously activated with low concentrations of cyanuric chloride (10.5 to 42.5 micromol/g gel). Well over 90% of the peptide used was immobilized. Three different concentrations were investigated for every immobilized tripeptide-ligand, 3.5, 7.0, and 14 micromol/g gel. The K(D) values of immobilized tripeptide-trypsin complexes were determined as well as the purifying performance and the trypsin-binding capacity of the affinity adsorbents. The K(D) values determined were in good agreement with the trypsin purification performance of the respective affinity adsorbents. The tripeptide sequence H-TPR-OH displayed the highest affinity for trypsin (K(D) 8.7 microM), whereas the sequence H-TAR-OH displayed the lowest (K(D) 38 microM). Dipeptide-ligands have failed to bind trypsin. When the ligand H-TPR-OH was immobilized via its N-terminal on agarose, at a concentration of 14 micromol/g gel, it produced the most effective affinity chromatography adsorbent. This adsorbent exhibited high trypsin-binding capacity (approximately 310,000 BAEE units/mL of adsorbent); furthermore, it purified trypsin from pancreatic crude extract to a specific activity of 15,200 BAEE units/mg (tenfold purification), and 82% yield. (c) 1997 John Wiley & Sons, Inc.