Unaffected Sites Research Articles

Secretion of arsenic, cholesterol, vitamin E, and zinc from the site of arsenical melanosis and leucomelanosis in skin

Background. Melanosis and leucomelanosis with or without keratosis are the earliest symptoms of arsenicosis. Uneven distribution of arsenical melanosis and leucomelanosis in skin led us to investigate the possibility of preferential secretion of arsenic and three constituents of sweat; cholesterol, vitamin E, and zinc. Methods. Twenty-four-hour skin secretion was collected from skin lesions and unaffected sites of 20 patients. Skin secretions were collected from 20 people exposed to arsenic-contaminated drinking water and 20 healthy, unexposed individuals. Results. The secretion of arsenic from the skin of healthy controls (mean ± SE; unit: μg/in.2 of skin/24 h; chest: 0.6 ± 0.2; back: 0.3 ± 0.1; abdomen: 0.5 ± 0.2) was increased several folds in arsenic-exposed controls (chest: 8.4 ± 1.8; back: 8.3 ± 1.9; abdomen: 6.7 ± 1.8) and patients (chest: 9.2 ± 1.3; back: 7.8 ± 1.3; abdomen: 5.2 ± 1.0). There was no difference in the skin lesions and unaffected sites in patients. However, the secretion of cholesterol was significantly lower in the chest of patients (190 ± 36) and healthy controls (686 ± 100) (p < 0.001). Secretions of vitamin E were low in healthy controls (chest: 8.5 ± 3.1; back: 5.2 ± 1.7; and abdomen: 8.7 ± 2.4), higher in arsenic-exposed controls (chest: 30.2 ± 8.1; back: 16.3 ± 8.9; and abdomen: 24.8 ± 9.3), and highest in patients [chest: 91.4 ± 14.9 (p < 0.0001 vs. control); back: 72.4 ± 13.2 (p < 0.001 vs. control); and abdomen: 46.8 ± 12.9]. Chronic intake of arsenic led to several folds higher secretion of zinc both in patients and in arsenic-exposed controls. One molecule of arsenic appears to be co-secreted with two molecules of zinc. Conclusion. Arsenic skin lesions showed no alteration in secretion of arsenic, although the secretion of cholesterol, vitamin E, and zinc was changed. Potential implications are discussed.

Additive effects from a combined treatment using parathyroid hormone and ibandronate in the ovariectomized rat

Additive effects from a combined treatment using parathyroid hormone and ibandronate in the ovariectomized rat

Arbuscular mycorrhiza of herbs colonizing a salt affected area near Kraków (Poland)

The arbuscular mycorrhizal (AM) status of plants colonizing an area affected by leakage of salty water (Barycz near Kraków, Poland) was studied in 2000 and 2001. The occurrence of plants typical for soils of increased salinity was observed. Among the 13 examined plant species 7 were mycorrhizal. The abundance of mycorrhizal plant populations was increased in the second year of study. Strains of 4 species of AMF, including &lt;em&gt;Glomus caledonium&lt;/em&gt;, &lt;em&gt;G. claroideum&lt;/em&gt;, &lt;em&gt;G. geosporum &lt;/em&gt;and &lt;em&gt;G. intraradices &lt;/em&gt;were isolated using trap cultures. On the basis of morphological characters the presence of &lt;em&gt;G. tenue &lt;/em&gt;was detected in plant roots of several species from the study area. The efficiency of mycorrhizal colonization and arbuscule formation by two strains of &lt;em&gt;G. geosporum &lt;/em&gt;isolated from a saline area and a strain of &lt;em&gt;G. intraradices &lt;/em&gt;from unaffected sites was tested in an experiment carried out on &lt;em&gt;Plantago lanceolata &lt;/em&gt;cultivated on substrata of different salinity levels. The increase in mycorrhizal parameters with growing salt content was observed in the case of strains originating from the salt-affected area. At the highest salt level these strains formed better developed mycorrhiza than the strain from the non-saline site, suggesting a better adaptation of the strains from the saline area. The data on vitality (alkaline phosphatase test) of intraradical AM fungi gave a clearer picture than those obtained by the conventional aniline blue staining.

Use of stable nitrogen isotope signatures of anthropogenic organic matter in the coastal environment: The case study of the Kosirina Bay (Murter Island, Croatia)

In this study stable nitrogen isotope ratios of particulate matter POM, zooplankton and selected biota such as Mytilus galloprovincialis were used to assess the impact of anthropogenically derived organic matter from the untreated domestic sewage, municipal and industrial effluents on the coastal ecosystem of the Kosirina Bay (Murter Island). The differences in δ15N values observed in POM and organisms collected in Kosirina Bay as compared to POM and biota sampled at unaffected sites from the southern part of the Kornati Island and highly impacted Pirovac Bay revealed only a very minor effects of anthropogenic inputs of nutrients and organic matter which is most probably derived from a sewage outfall south of the Tužbina Island.

Marker Paper, HMP, skin

The Human Microbiome Project seeks to explore the diversity of microbiota that resides in and on the human body, including the skin, in health and disease. The NIH Intramural Skin Microbiome Consortium (NISMC) is a trans-disciplinary group of experts engaged in the practice of genomics, bioinformatics, large-scale DNA sequencing, dermatology, immunology, allergy, infectious disease, and clinical microbiology. Atopic dermatitis (AD, “eczema”) is a chronic relapsing skin disorder that affects ~15% of U.S. children and is associated with $1 billion of medical costs annually. AD is characterized by dry, itchy skin, infiltrated with immune cells. Colonization by _Staphylococcus aureus_ (S. aureus) is ten-fold more common in AD patients and is associated with disease flares. We hypothesize that, in addition to S. aureus, AD may also be associated with additional microbes and/or selective shifts of commensal microbes that are relevant to disease progression. The NISMC seeks to define the microbiota that resides in and on the skin and nares of three patient groups, all of whom have eczematous lesions and are currently seen at the NIH Clinical Center: (1) AD patients; (2) Wiskott-Aldrich syndrome (WAS) patients; and (3) Hyper IgE syndrome (HIES) syndrome patients.Examination of the microbiome of patients with WAS or HIES syndromes, both rare immunodeficiencies, will advance our understanding of how an individual’s immune system shapes their cutaneous microbial community. We are performing a prospective longitudinal study that follows these groups of patient thorough the cycles of eczema flares, ascertaining clinical data and samples at each stage. To analyze the longitudinal samples obtained from both affected and unaffected skin sites, we will use an integrated approach including: (1) extensive characterization of bacterial diversity; (2) analysis of fungal diversity; (3) generation of whole-genome sequences of novel skin microbial isolates; and (4) metagenomic sequencing to assess microbial abundance and deduce metabolic activities. Examination of the microbial communities on affected and unaffected skin of patients with eczema will yield insights into the gene-environment interactions relevant to these diseases.

Skin Microbiome in Disease States: Atopic Dermatitis and Immunodeficiency

AbstractThe Human Microbiome Project seeks to explore the diversity of microbiota that resides in and on the human body, including the skin, in health and disease. The NIH Intramural Skin Microbiome Consortium (NISMC) is a trans-disciplinary group of experts engaged in the practice of genomics, bioinformatics, large-scale DNA sequencing, dermatology, immunology, allergy, infectious disease, and clinical microbiology. Atopic dermatitis (AD, “eczema”) is a chronic relapsing skin disorder that affects ~15% of U.S. children and is associated with $1 billion of medical costs annually. AD is characterized by dry, itchy skin, infiltrated with immune cells. Colonization by _Staphylococcus aureus_ (S. aureus) is ten-fold more common in AD patients and is associated with disease flares. We hypothesize that, in addition to S. aureus, AD may also be associated with additional microbes and/or selective shifts of commensal microbes that are relevant to disease progression. The NISMC seeks to define the microbiota that resides in and on the skin and nares of three patient groups, all of whom have eczematous lesions and are currently seen at the NIH Clinical Center: (1) AD patients; (2) Wiskott-Aldrich syndrome (WAS) patients; and (3) Hyper IgE syndrome (HIES) syndrome patients.Examination of the microbiome of patients with WAS or HIES syndromes, both rare immunodeficiencies, will advance our understanding of how an individual’s immune system shapes their cutaneous microbial community. We are performing a prospective longitudinal study that follows these groups of patient thorough the cycles of eczema flares, ascertaining clinical data and samples at each stage. To analyze the longitudinal samples obtained from both affected and unaffected skin sites, we will use an integrated approach including: (1) extensive characterization of bacterial diversity; (2) analysis of fungal diversity; (3) generation of whole-genome sequences of novel skin microbial isolates; and (4) metagenomic sequencing to assess microbial abundance and deduce metabolic activities. Examination of the microbial communities on affected and unaffected skin of patients with eczema will yield insights into the gene-environment interactions relevant to these diseases.

Erythema multiforme, Stevens–Johnson syndrome and toxic epidermal necrolysis: Frozen‐section diagnosis

Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) may be fatal. Although classified by body surface area skin detachment, initial stages of both may present with erythema multiforme (EM)-like lesions. To diagnose and predict disease activity adequately as early as possible for patients revealing EM-like lesions, we performed frozen-section diagnosis. Thirty-five patients clinically diagnosed as EM, SJS or TEN were biopsied to diagnose and predict disease progression within the initial-visit day. Half of a histological section taken from a lesion was snap-frozen and immediately cryostat-sectioned, acetone-fixed and stained with hematoxylin-eosin. Specimens were examined with light microscopy for presence of epidermal necrosis. A section from unaffected sites was also examined for 11 patients. Specimens were examined with light microscopy for presence of graft-versus-host reaction (GVHR)-like findings: apoptotic keratinocytes and satellite cell necrosis. Epidermal necrosis was seen in nine patients. Initial diagnosis of the nine was one of overlap SJS-TEN, four of SJS and four of EM, and final diagnosis of those was one of TEN, one of overlap SJS-TEN, four of SJS and three of EM. Dissociation between initial and final diagnosis was seen in three cases. GVHR-like findings in the epidermis were observed in two patients finally diagnosed as overlap SJS-TEN and TEN. Frozen sections are useful not only to make a diagnosis of erythema multiforme but to assess a potential to exhibit more aggressive clinical behaviors (SJS or TEN).

Impact of severe forest dieback caused by Phytophthora cinnamomi on macrofungal diversity in the northern jarrah forest of Western Australia

Abstract The relative diversity and abundance of different functional groups of macrofungi were investigated in the northern jarrah forest, a mediterranean climate sclerophyllous forest dominated by eucalyptus trees in Western Australia. We sampled paired sites that were either severely affected by dieback, a disease caused by Phytophthora cinnamomi which causes selective plant mortality, or unaffected by this type of forest decline. Macrofungi were sampled 3 times during the growing season along six 100 m × 2 m transects in these sites. Dieback-unaffected sites were found to have significantly different macrofungal floras than unaffected sites. Macrofungal abundance and diversity were approximately 1.5 times and 1.8 times greater respectively in dieback-unaffected sites than in severely affected sites. Dieback-affected sites had a similar diversity of saprotrophic and ectomycorrhizal fungi, whereas more fungal taxa on dieback-unaffected sites were mycorrhizal (>60%). Dung fungi were the most common saprophytes, especially in dieback-affected sites, but abundance data greatly overestimated the importance of these relatively small fungi. We concluded that vegetation changes linked to dieback had a negative effect on fungal community structure and biodiversity in the northern jarrah forest, in a similar manner to other forms of severe disturbance. Conversely, high tree mortality increased the abundance of wood decay fungi, at least in the short term. We expect that reductions in macrofungal species richness were indirectly linked to impacts on mycorrhizal host plants and saprotrophic substrates. Our data show that changes in vegetation composition had the greatest effect on ectomycorrhizal fungi, presumably due to their obligate symbiotic associations.

Plant-metal (Al, Cd, Fe, Mn, Mo, Pb, Zn) interactions in contaminated freshwater sediments of the Fly River Floodplain, Papua New Guinea.

The uptake of Al, Cd, Fe, Mn, Mo, Pb and Zn by 23 aquatic macrophytes species from mine-impacted sediments from the Fly River flood plain in Papua New Guinea was investigated by analysis of sediment, water and plant samples. Increased metal concentrations in aquatic plants were observed at sites contaminated by mine-derived sediment. The study also showed that the plant metals were primarily derived from the sediment, except molybdenum, which was also correlated to it dissolved species. The factor increase ranges for Cd (2-9), Fe (2-95), Mn (2-174), Pb (2-10), Zn (2-79, Al (2-7) and Mo (2-29) were calculated relative to background concentrations. These factors increased with the degree of impact from mire-derived sediment. Based on analysis of the data presented in this study the monocots L. Hexandra, H. Acutigluma and I. Polystachium could be targeted for further study. Only I. Polystachium appear to be the best choice for monitoring of the impact of mine-derived sediment on aquatic plant species in the Fly River. Leaves generally have higher metal concentration than stems in monocots. Of the two dicots which were compared, L. Hyssopifolia displayed a trend common to all sites, where metal concentrations decrease in the following pattern seeds<stems<shoots<leaves. Thie following trend was observed for N. Nucifera leaves<stems<flowers<pods<seeds for all elements except aluminium and manganese. The opposite was observed for Fe and Pb in N. Nucifera at the control site. Fully submerged species were found to contain metal concentrations that were higher, sometimes orders fo magnitude higher compared with emergent species. Submerged species from unaffected sites still had higher concentrations compared to emergent species from contaminated sites. Plant metal concentrations are highly dependent on the Eh, pH and organic matter.teh significant negative relationship between SOM and plant metals (with Cd, Fe, Al and Mo) reveal that these metals are bound up as organic matter complex. However, these can then become available as the organic matter complexes breakdown during oxidation .Similarly, under reduced conditions most metals are bound up as metal sulphides, however changes in the redox status of the sediment from reducing to oxidising also renders these metals available for uptake.

Development and validation of a multiplex bead assay for measuring growth mediators in wound fluid

Large amounts of biological samples are usually required to measure multiple components by the enzyme-linked immunosorbant assay. However, the amounts of many tissue extracts and fluids, including gingival crevicular fluid (GCF), are generally extremely small. The aim of this study was, therefore, to develop and validate a novel multiplex bead assay (MBA) to simultaneously measure a profile of healing-related mediators in the GCF of treated periodontal wounds. An MBA was developed and validated by assessment of assay selectivity, recovery, precision and sensitivity, using eight recombinant human growth mediators as assay standards. GCF samples were collected on paper strips from healing wound (test) and healthy unaffected (control) sites of 15 patients with periodontitis, seven days post-periodontal surgery. Each GCF sample was eluted and the levels of the mediators measured using the MBA and antibody pairs specific for angiopoietin-1, vascular endothelial growth-factor, bone morphogenetic protein-2, osteoprotegerin, tissue inhibitor of metalloprotease-1 (TIMP-1), basic fibroblast growth-factor, keratinocyte growth-factor, and platelet derived growth-factor. Less than 1.8% of cross-reactivity was observed between antibodies and the eight different analytes, for which the recovery was more than 85%. Mean intra- and inter-assay precision were within the acceptance criteria of 20% and 25%, respectively. Detection of all mediators was highly sensitive (<or=70 ng/L) except for TIMP-1 (215 ng/L). Angiogenic factors were the most highly secreted in the GCF seven days post-surgery. This new MBA can simultaneously measure small amounts of eight different growth mediators in the GCF of healing periodontal wounds. It might also be a valuable tool for evaluating the components of wound fluids as a prognostic indicator of the success of therapeutic intervention.

The impact of former iron ore mining on the transformation of vegetation cover of the Gielniowski Hump (Małopolska Upland)

The impact of former iron ore mining on the transformation of vegetation cover of the Gielniowski Hump (Małopolska Upland) The impact of the 17th-19th century iron ore mining on the current state of vegetation cover of the Gielniowski Hump, an upland region situated within the Old Polish Industrial Region (a former centre of mining and smelting), was studied. In the course of the research, the detailed floristic lists were compiled using the cartogram method (ATPOL grid) and phytosociological relevés were made according to the Braun-Blanquet method. Relevés were taken both at the post-mining sites (on spoil banks) and, for comparative purposes, in the adjacent, not affected areas. The obtained results show significant differences between the flora and vegetation of the sites affected and unaffected by mining. These differences result from the changed mineralogical composition of soil in the post-mining areas, which has lead to an increase in the proportion of mesophilous forest communities and in the number of species of the class Querco-Fagetea as compared to the unaffected sites. Overall, the former iron ore mining activity has increased the biodiversity level in the Gielniowski Hump mesoregion due to an increase in the total number of species in the area, brought about by the formation of a mining-related habitat mosaic.

Material properties are related to stress fracture callus and porosity of cortical bone tissue at affected and unaffected sites

Stress fractures are overuse injuries of bone that affect elite athletes and military recruits. One response of cortical bone to stress fracture is to lay down periosteal callus. The objectives of this study were to determine if material properties are different among bones with different stages of stress fracture callus, at both a callus site and at a distal site. Cortical specimens were mechanically tested to determine their stress-strain response. Material property differences were examined using nonparametric and regression analyses. At the callus site, material properties were low during the earliest stages of callus, higher with increasing callus maturity, but dropped at the late stage of callus. At the distal site, the material properties were low during early stages of callus and approached, or returned to, those of bones without callus during the late stages of callus. The effects of stress fracture and bone callus are not limited to the focal site of stress fracture.

Species composition and diversity of aero-terrestrial algae and cyanobacteria of the Boreč Hill ventaroles.

The algal flora of the Boreč Hill ventaroles was examined and compared with the flora of their close surroundings. In comparison to unaffected sites, the ventaroles differ in seasonal temperature fluctuation as well as in soil pH. Winter exhalations caused by continual air circulation in the cranny system of rock massive result in a significant increase in air temperature and soil pH. The ventaroles were inhabited by a markedly richer and more diversified algal flora. Over half of investigated species occurred only in the ventaroles, including all chrysophyte, eustigmatophyte and desmid taxa. In contrast to the unaffected sites, different algal populations were discovered in the ventaroles. The investigated species could be separated into two groups: ventarole-specific ones preferentially occurred in both the ventaroles, and those occurred only in a large ventarole. There, water condensation in moss plants during winter exhalations enables a short-term occurrence of several algal species preferring aquatic environments. The diatom flora of the ventaroles resembles well the species composition found in the caves. By contrast, the desmid flora is rather similar to algal communities found in ephemeral water bodieslike temporary peat bog pools or dripping rocks. Thus, the Boreč Hill ventaroles represent a unique type of biotope, with a specific algal flora adapted to periodical periods of warm, moist air exhalations of several months' duration.

Age-Dependent Demise of GNAS-Mutated Skeletal Stem Cells and “Normalization” of Fibrous Dysplasia of Bone

We studied the role of somatic mosaicism in fibrous dysplasia of bone (FD) within the context of skeletal ("mesenchymal") stem cells by assessing the frequency of mutated colony forming unit-fibroblasts (CFU-Fs) from FD lesions, and in some cases, from unaffected sites, in a series of patients. There was a tight inverse correlation between the percentage mutant CFU-F versus age, suggesting demise of mutant stem cells caused by exuberant apoptosis noted in samples from young patients. In older patients, either partially or completely normal bone/marrow histology was observed. On in vivo transplantation, FD ossicles were generated only by cell strains in which mutant CFU-Fs were identified. Strains that lacked mutant CFU-F (but were mutation positive) failed to regenerate an FD ossicle. These data indicate that GNAS mutations are only pathogenic when in clonogenic skeletal stem cells. From these data, we have evolved the novel concept of "normalization" of FD. As a lesion ages, mutant stem cells fail to self-renew, and their progeny are consumed by apoptosis, whereas residual normal stem cells survive, self-renew, and enable formation of a normal structure. This suggests that activating GNAS mutations disrupt a pathway that is required for skeletal stem cell self-renewal.

Quantitative phosphoproteome analysis of a mouse liver cell line reveals specificity of phosphatase inhibitors

The liver is a central organ involved in many aspects of physiology and disease. Signaling properties of hepatocytes, the main liver cell type, are of special interest in metabolic diseases and in regeneration. For this reason we investigated the phosphoproteome of the mouse liver cell line Hepa1-6 by stable isotope labeling by amino acids in cell culture (SILAC) and high resolution MS. Using stringent statistical evaluation criteria, we obtained 5433 phosphorylation sites on 1808 proteins. The phosphoproteome encompasses all major protein classes, including a large number of transcription factors. We compared control and phosphatase inhibitor treated cells by SILAC. This enabled ready identification of in vivo phosphorylation sites by sequencing the more abundant, inhibitor induced version of the peptide while still observing the endogenous version. We employed a mixture of pervanadate for blocking protein tyrosine phosphatases (PTPs) and calyculin A and deltamethrin for blocking the activities of serine/threonine phosphatases. Interestingly, these commonly used inhibitors in standard concentrations affected only 28% of the phosphopeptides by at least two-fold. The unaffected sites may be substrates of phosphatases that are not efficiently inhibited, have slow kinetic or sites that are almost stoichiometric in normally growing cells. Finally, we devised a triple labeling strategy comprising control cells, stimulated cells, and phosphatase treated cells to derive an upper bound on phosphorylation occupancy.

Feather-picking Psittacines: Histopathology and Species Trends

Histologic findings are described for 408 feather-picking or self-mutilating psittacines with the use of biopsies from clinically affected and unaffected skin. Inflammatory skin disease was diagnosed in 210 birds, and traumatic skin disease was diagnosed in 198 birds. Criteria used for the diagnosis of inflammatory skin disease included the presence of perivascular inflammation in the superficial or deep dermis of clinically affected and unaffected sites. The primary histologic criteria for the diagnosis of traumatic skin disease were superficial dermal scarring with or without inflammation in the affected sites and an absence of inflammation in the unaffected sites. The inflammatory cells associated with the lesions were typically lymphocytes and occasionally plasma cells, histiocytes, and granulocytes. A preponderance of inflammatory skin disease was seen in macaws (Ara spp.) and Amazon parrots (Amazona spp.). A preponderance of traumatic skin disease was seen in cockatoos (Cacatua spp.) and African grey parrots (Psittacus erithacus). The prevalence of each was approximately equal in several other species, including conures (Aratinga and Pyrrhura spp.), eclectus parrots (Eclectus roratus), quaker parrots (Myiopsitta monachus), cockatiels (Nymphicus hollandicus), parakeets (Cyanorhamphus and Psittacula spp.), and caiques (Pionites spp.). No geographic or gender-based trends were identified. These findings could be helpful for identifying and treating birds with feather-picking disorders.

Bivalve tissue as a carbon and nitrogen isotope baseline indicator in coastal ecosystems

Pinctada fucata martensii mantle tissue and gut contents were examined as baseline indicators of carbon and nitrogen isotope composition at six stations in the Uwa Sea, Japan. Substantial variations in δ 13C and δ 15N values of oysters among stations were observed, with δ 13C being consistently lower at Hiburi Island (−18.1‰) than at other stations (−17.2‰). Oysters from fish farm sites were enriched in δ 15N (8.1‰) relative to those from unaffected sites (6.8‰), suggesting that fish farming tends to increase baseline δ 15N values. The mean Δ δ 13C (0.8‰) was consistent over space and time, whereas the average Δ δ 15N slightly increased in summer. The relatively low δ 15N enrichment compared to the theoretical isotope fractionation factor (3.4‰) may be due to oyster-specific physiological attributes. Carbon and nitrogen isotope turnover rates were roughly similar within a tissue, and mantle tissue turnover rate was estimated to be 120–180 days. These results indicated that oysters are long-term integrators of δ 13C and δ 15N from their diet and that δ 13C of oysters is a more accurate bioindicator of isotopic baselines than δ 15N for marine ecological studies.

Involvement of innate immunity in the pathogenesis of intestinal Behçet's disease

The involvement of excessive T helper 1 (Th1) cell functions in the pathogenesis of Behçet's disease (BD) has been reported. We therefore studied Toll-like receptor (TLR)-expressing cells, which play important roles in innate immunity in patients with BD. Peripheral blood mononuclear cells (PBMC) of BD and healthy controls, and tissue specimens of intestinal BD and Crohn's disease (CD) were analysed for messenger RNA (mRNA) and protein expressions by reverse transcription-polymerase chain reaction and immunostaining respectively. PBMC of BD expressed TLR-2 and TLR-4 mRNA almost comparable with healthy controls. Intestinal lesions of BD expressed TLR-2 and TLR-4 mRNA consistently. In contrast, TLR-4 mRNA was expressed preferentially and TLR-2 mRNA was expressed less frequently in CD lesions. In intestinal samples of BD, TLR-2 and TLR-4 mRNA were detected in ileocaecal ulcer lesions, but not in unaffected sites of the same sample, indicating the association of the TLR expression with the disease manifestation of intestinal BD. TLR-2-expressing cells which were simultaneously cluster of distribution (CD)68-positive produced interleukin (IL)-12 in the lesions, indicating the participation of TLR-2-expressing cells in the Th1 skewed responses in vivo. As a possible ligand of TLR-2, in BD self-heat shock protein 60 was expressed in peripheral blood lymphocytes and intestinal tissues. Collectively, TLR-2-expressing cells as well as TLR-4-expressing cells accumulated in the intestinal lesions of BD. IL-12 produced by TLR-2-expressing cells may contribute to the induction of Th1-dominant immune responses in intestinal BD.

Impact of fish farming on the distribution of phosphorus in sediments in the middle Adriatic area

During the last decade, intensive fish farming developed along the central Croatian coast, creating a need to study and evaluate its potential influence on unaffected sites. We considered phosphorus as an indicator of the influence of fish farming and investigated the distribution of phosphorus forms in sediment from several fish farms and marine areas of different trophic status in the middle Adriatic. Analyses of samples were performed with modified SEDEX techniques. Our results indicated that authigenic apatite phosphorus showed no significant differences among the investigated stations, while organic phosphorus concentrations reflected the trophic status of the station area. Below-cage sediment was characterized by enhanced fish debris phosphorus and low detrital apatite phosphorus concentrations, while sediment from an anthropogenically influenced bay showed the highest values of iron bound phosphorus species. Among the different P fractions, fish debris phosphorus proved to be the most sensitive indicator of the influence of fish farming on marine sediment.

Relationships Among Exceedences of Sediment Guidelines, the Results of Ambient Sediment Toxicity Tests, and Community Metrics in Estuarine Systems

To use bioassessments to help diagnose or identify the specific environmental stressors affecting estuaries, we need a better understanding of the relationships among sediment chemistry guidelines, ambient toxicity tests, and community metrics. However, this relationship is not simple because metrics generally assess the responses at the community level of biological organization whereas sediment guidelines and ambient toxicity tests generally assess or are based on the responses at the organism level. The relationship may be further complicated by the influence of other chemical and physical variables that affect the bioavailability and toxicity of chemical contaminants in the environment. Between 1990 and 1993, the U.S. Environmental Protection Agency (USEPA) conducted an Environmental Monitoring and Assessment Program (EMAP) survey of estuarine sites in the Virginian Province of the eastern United States. The surveys collected data on benthic assemblages, physical and chemical habitat characteristics, and sediment chemistry and toxicity. We characterized these estuarine sites as affected by sediment contamination based on the exceedence of sediment guidelines or on ambient sediment toxicity tests (i.e., 10-day Ampelisca abdita survival). Then, benthic invertebrate metrics were compared among affected and unaffected sites to identify metrics sensitive to the contamination. A number of benthic invertebrate metrics differed between groups of sites segregated using the organism-level measures whereas other metrics did not. The difference among metrics appears to depend on the sensitivity of the individual metrics to the stressor gradient represented by metals or persistent organic toxics in sediments because the insensitive metrics do not effectively quantify the changes in the benthic invertebrate assemblage associated with these stressors. The significant relationships suggest that a relationship exists between the organism-level effects assessed by chemistry or ambient toxicity tests and the community-level effects assessed by community metrics and that the organism-level effects are predictive, to some extent, of community-level effects.