Floating plastics as integrative samplers of organic contaminants of legacy and emerging concern from Western Mediterranean coastal areas.

This study investigates the role of floating plastics as integrative samplers of organic contaminants. To this end, plastics items were collected in two Western Mediterranean coastal areas: the Mar Menor lagoon, and the last transect of Ebro river. Floating plastics were identified and characterized by attenuated total reflection Fourier-transform infrared spectrometry. Then, organic contaminants were extracted from plastic items by ultrasonic extraction with methanol, and the concentrations of 168 regulated and emerging contaminants were analysed. These compounds were analysed by stir bar sorptive extraction coupled to gas chromatography-mass spectrometry (GC-MS), except for bisphenol analogues, which were analysed with a ultraperformance liquid chromatography pump coupled to a triple quadrupole mass spectrometer (UHPLC-MS/MS), and pharmaceutical compounds, determined by UPLC coupled to hybrid triple quadrupole-linear ion trap mass spectrometer (UPLC-MS/MS). All the contaminants groups considered were detected in the samples, being particularly relevant the contribution of plastic additives. The most frequently detected contaminants were UV-filters, PAHs, pharmaceuticals and synthetic musks. Apart from plasticizers, the individual contaminants octocrylene, homosalate, galaxolide, salycilic acid and ketoprofen were frequently detected in plastics items. The results pointed out to urban and touristic activities as the main sources of pollution in the coastal areas investigated. The utility of floating plastics as integrative samplers for the detection of organic contaminants in aquatic ecosystems has been demonstrated.

Combined exposure of the bivalve Mytilus galloprovincialis to polyethylene microplastics and two pharmaceuticals (citalopram and bezafibrate): Bioaccumulation and metabolomic studies.

Pharmaceuticals and microplastics constitute potential hazards in aquatic systems, but their combined effects and underlying toxicity mechanisms remain largely unknown. In this study, a simultaneous characterization of bioaccumulation, associated metabolomic alterations and potential recovery mechanisms was performed. Specifically, a bioassay on Mediterranean mussels (Mytilus galloprovincialis) was carried out with polyethylene microplastics (PE-MPLs, 1 mg/L) and citalopram or bezafibrate (500 ng/L). Single and co-exposure scenarios lasted 21 days, followed by a 7-day depuration period to assess their potential recovery. PE-MPLs delayed the bioaccumulation of citalopram (lower mean at 10 d: 447 compared to 770 ng/g dw under single exposure), although reaching similar tissue concentrations after 21 d. A more limited accumulation of bezafibrate was observed overall, regardless of PE-MPLs co-exposure (

Occurrence and distribution of contaminants of legacy and emerging concern in surface waters of two Western Mediterranean coastal areas: Mar Menor Lagoon and Ebro Delta.

The occurrence and distribution of UV filters, plastic additives, synthetic musks, other personal care products (Other PCPs), triazines, polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs), organophosphorus pesticides (OPPs), polychlorinated biphenyls (PCBs) and other current-use pesticides (Other CUPs) were characterised during summer 2018 and winter 2019 in surface waters of two sensitive areas of the Spanish coast located on the Mediterranean Sea (Mar Menor lagoon and Ebro Delta). Sixty-three organic contaminants out of a total of 100 compounds were detected, thus confirming the presence of all groups of pollutants studied in surface water at concentrations of ng/L. Both areas are affected by agricultural, urban and recreational activities, PCPs (mainly UV filters) being the predominant compounds found in both seasons which showed significant increases in concentrations in summer. The contaminants found at the highest concentrations were octocrylene, homosalate and ethylhexyl salicylate, which showed risk quotients higher than 1, indicating a potential risk to aquatic organisms, particularly in summer.

Occurrence of selected endocrine disrupting compounds in Iberian coastal areas and assessment of the environmental risk.

The spatial and temporal distribution of selected endocrine disrupting compounds (4-tert-octylphenol, 4-n-octylphenol, 4-n-nonylphenol, nonylphenol, and bisphenol A) in two coastal areas of the Iberian Peninsula (Ria de Vigo and Mar Menor lagoon) were evaluated for the first time. Seawater and sediment samples collected during spring and autumn of 2015 were analysed using greener extraction techniques and liquid chromatography-tandem mass spectrometry. The presence of branched isomers (4-tert-octylphenol and nonylphenol) and bisphenol A in almost all seawater and sediment samples demonstrated their importance as pollutants in the frame of water policy, while no concentrations of linear isomers (4-n-octylphenol and 4-n-nonylphenol) were found. Higher seawater levels were observed in Mar Menor lagoon, especially in spring, associated with wastewater treatment plant effluents and nautical, agricultural and industrial activities. Similar sediment concentrations were measured in both studied areas, being nonylphenol levels five times higher than those measured for the other EDCs. Experimental sediment-water partition coefficients showed a moderate sorption of target compounds to sediments. Risk quotients for water compartment evidenced a moderate risk posed by nonylphenol, considering the worst-case scenario. For sediments, moderate risk related to 4-tert-octylphenol and high risk to nonylphenol were estimated.

Impact of flash flood events on the distribution of organic pollutants in surface sediments from a Mediterranean coastal lagoon (Mar Menor, SE Spain).

The influence of flash flood events on the input and distribution of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and organochlorinated pesticides (OCPs) in surface sediments from the Mar Menor lagoon were characterized in this study. These contaminants were analyzed in surface water samples collected during two flash flood events in the main surface watercourse which flow into the Mar Menor lagoon. Surface sediments were sampled semiannually before and after flash flood events. The total input of PAHs, OCPs, and PCBs (sorbed + dissolved) during two flash flood events was estimated at 0.98, 1.32, and 0.34 kg, respectively, the main input corresponding to p,p'-DDE (1.00 kg). The distribution of organic contaminants in surface sediments was not homogeneous as a consequence of the presence of many simultaneous sources and different meteorological, hydrodynamic, and physicochemical conditions. As a consequence of flash flood events, p,p'-DDE concentrations in surface sediments increased significantly in the central and south zones of the lagoon. However, in the case of PCBs, a dilution effect was observed in the south zone after such events, reducing the environmental risk. These changes in the pollutant distribution persisted at least 1 year later (autumn 2010), showing that the impact of flood events in the distribution of persistent organic contaminants in Mediterranean coastal lagoons is of relevance according to the ecological risk assessment carried out. The impact of these events should be also considered in other coastal systems, especially in semiarid and semiconfined areas.

Pretransplant CD28 Biomarker (Levels of Expression and Quantification of Molecules per Cell) in Peripheral CD4+ T Cells Predicts Acute Rejection Episodes in Liver and Kidney Recipients.

BACKGROUND: Acute rejection (AR) remains a significant cause of graft loss. Better approaches to predict AR are being investigated. Surface CD28 protein is essential for T-cell proliferation and survival as well as cytokine production. PATIENTS AND METHODS: Pretransplant CD4+CD28+ peripheral T cells were examined in 30 liver recipients (LRs) and 31 kidney recipients (KRs) by flow cytometry. RESULTS: Pretransplant CD4+CD28+ T cells in LRs were significantly lower in rejectors than nonrejectors (P = .002). Furthermore, the total number of CD28 molecules per cell in LRs (P = .02) as well as KRs (P = .047) was significantly lower in rejectors than nonrejectors. The healthy group did not display differences when compared with patients with end-stage liver disease or renal failure; however, stratification analysis displayed higher levels of CD4+CD28+ when compared with rejected LRs (P = .04) but not KRs. CD28 levels <41.94% were able to discriminate LRs at high risk of AR (P = .003). Similarly, a total number of CD28 molecules ≤8359 (P = .031) in LRs and ≤7669 (P = .046) in KRs correlated with high risk of AR. CONCLUSION: The preliminary results presented herein exhibit a fast and noninvasive method that assists clinicians to prevent AR by monitoring CD4+CD28+ peripheral T cells.

Report From the First and Second Spanish Killer Immunoglobulin-Like Receptor Genotyping Workshops: External Quality Control for Natural Killer Alloreactive Donor Selection in Haploidentical Stem Cell Transplantation.

An important factor affecting the success in the setting of related haploidentical hematopoietic stem cell transplantation (HSCT) is the graft-versus-leukemia effect mediated by natural killer (NK) cells when the donor displays NK alloreactivity versus the recipient. NK cell function is regulated by killer immunoglobulin-like receptors (KIR) and it has been described that donor KIR genotype influences transplantation outcome. This has led to a requirement of laboratories to have a quality assurance program for validation and control of their KIR genotyping methods. The goal of the 1st and 2nd Spanish KIR Genotyping Workshops was to provide an external proficiency testing program in KIR genotyping for Spanish immunology and transplant laboratories. These workshops were conducted during the years 2014-2016 and consisted of 17 participating laboratories typing a set of 20 samples. The presence/absence of 16 mandatory KIR loci (2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 2DP1, 3DL1, 3DL2, 3DL3, 3DS1, and 3DP1) was evaluated per sample. Methods for KIR genotyping included polymerase chain reaction with the use of sequence-specific primers and sequence-specific oligoprobes. Consensus typing was reached in all samples, and the performance of laboratories in external proficiency testing was satisfactory in all cases. The polymorphism detected in the small sample studied in both workshops is indicative of an ample variety of KIR gene profiles in the Spanish population.

Pro- and anti-inflammatory cytokine gene single-nucleotide polymorphisms in inflammatory bowel disease.

Anti-inflammatory cytokines have an important role in disease, tumour and transplant processes. Alterations in the regulation of several cytokines have been implicated in a variety of inflammatory disorders, including IBD (inflammatory bowel disease) [Crohn's disease (CD) and ulcerative colitis (UC)]. Cytokine polymorphisms are also known to affect the level of gene expression. Thus, the aim of this study was to determine the relationship between cytokine polymorphisms and the IBD pathologies in a Spanish population. Polymorphisms analysis was performed using PCR-SSOP using a microbeads luminex assay. The following polymorphisms were determined: TNFα [-238G/A (rs361525) and -308G/A (rs1800629)], IFNγ [+874A/T (rs62559044)], TGFß [+869C/T (rs1982073) and +915G/C (rs1800471)], IL10 [-1082A/A (rs1800896), -592A/C (rs1800872), -819C/T (rs1800871)], IL6 [-174C/G (rs1800795)], IL12p40 [3'UTR -1188A/C (rs3212227)], IL1α [-889C/T (rs1800587)], IL1ß [-511C/T (rs1143634) and +3962C/T (rs1143633)], IL1R [Pst-1 1970C/T] and IL1RA [Mspa-1 11100C/T]. No statistical differences in TNFα, IFNγ, TGFß, IL10, IL6, IL1α, IL1ß, IL1R and IL1Ra genotypes and allele distributions between the IBD groups and healthy controls were found. However, we observed significant differences in the 3'UTR -1188A/C polymorphism of IL12p40. So -1188A allele was increased in patients with UC and the -1188C allele (high IL12p40 production) was increased in patients with CD with respect to controls. These data are in concordance with the fact that CD has been shown to be associated with a Th1 T-cell-mediated inflammation model and high IL12/IFNγ production at histological affected sites. These data suggest that cytokine polymorphisms in TNFα, IFNγ, TGFß, IL10, IL6 and IL1α, IL1ß, IL1R and IL1Ra cytokine gene do not seem to be relevant in IBD susceptibility and IL12p40 3'UTR -1188A/C polymorphism seems to be associated with a differential IBD development.

Heterogeneous distribution of polycyclic aromatic hydrocarbons in surface sediments and red mullet along the Spanish Mediterranean coast.

The spatial distribution of polycyclic aromatic hydrocarbon (PAH) concentrations was characterised in surface sediments and red mullet from eleven Iberian Mediterranean coastal areas. Mean PAH concentrations ranged from 28 to 1006 and from 3 to 40 µg kg(-1)d.w. in sediment and red mullet muscle, respectively. The highest PAH concentrations in sediments were detected close to main ports and urban nuclei. However, concentrations of PAHs in red mullet showed little correspondence with PAH concentrations of the sediments due to its metabolic capacity. Phenanthrene was the predominant homologue in red mullet, whereas fluoranthene, pyrene and benzo(b)fluoranthene were the most abundant ones in sediments. Significant correlations between PAHs and organic carbon or fine fraction in sediments were only found in some areas. PAH concentrations in sediments were lower than environmental criteria in the majority of cases, except for benzo(g,h,i)perylene in 25% of samples from the Barcelona coastal area and for several homologues close to the port of Sagunto.

Genetic polymorphisms of tumour necrosis factor alpha (TNF-α) promoter gene and response to TNF-α inhibitors in Spanish patients with inflammatory bowel disease.

Tumour necrosis factor alpha (TNF-α) has an important role in inflammatory response. Alterations in the regulation of TNF-α have been implicated in a variety of inflammatory disorders, including Inflammatory bowel disease (IBD). Indeed, a common treatment for IBD is the use of TNF-α inhibitors. Polymorphisms in the TNF-α promoter region are known to affect the level of gene expression. Our aim was to investigate the influence of these single nucleotide polymorphisms (SNPs) in TNF-α promoter gene play in the risk of IBD in a Spanish population and their individual response to anti-TNF-α treatment. DNA samples from patients with IBD and controls were screened for TNF-α -238G/A (rs361525) and -308G/A (rs1800629) SNPs by PCR-SSOP using a microbeads luminex assay and compared with response to TNF-α inhibitors. There were not statistical differences in -238G/A and -308G/A allele and genotype frequencies between patients. However, we found an increased frequency of -308A allele and -308GA genotype in these nonresponders patients to TNF-α inhibitors with respect to responders patients (Pc < 0.05). This -308GA genotype has been classified as high producer of this cytokine. This fact could actually be interesting to explain the different response of patients with IBD with respect to TNF-α inhibitors. TNF-α promoter gene polymorphism does not seem to play a role in IBD susceptibility, but particular TNF-α genotypes may be involved in the different responses to TNF-α inhibitor treatment in Spanish patients with IBD.

Influence of an intensive agricultural drainage basin on the seasonal distribution of organic pollutants in seawater from a Mediterranean coastal lagoon (Mar Menor, SE Spain).

The seasonal distribution of polycyclic aromatic hydrocarbons, polychlorinated biphenyls, organochlorine pesticides, organophosphorus pesticides, triazines and other organic pollutants in surface seawater from the Mar Menor lagoon (SE Spain) was characterized from spring 2009 to winter 2010 by stir bar sorptive extraction and thermal desorption followed by capillary gas chromatography coupled to mass spectrometry (SBSE-GC-MS). The most ubiquitous pollutants were chlorpyrifos, chlortal-dimethyl, terbuthylazine, naphthalene and propyzamide throughout the year. Insecticides and herbicides were predominant in summer and spring, and fungicides were more abundant in autumn and winter. Chlorpyrifos was predominant in autumn and spring; herbicides (propyzamide, terbuthylazine and terbuthylazine-desethyl) in autumn and fungicides (flutolanil, boscalid, cyprodinil) in autumn and winter. The highest concentrations for the majority of pollutants were detected in autumn as consequence of air and surface/ground water inputs, and above all of desorption from terrigenous materials deposited in sediments by two intense flash flood events (September 2009).

Complement component C6 deficiency in a Spanish family: implications for clinical and molecular diagnosis.

Complement component C6 deficiency is a genetic disease presenting as increased susceptibility to invasive Neisseria meningitidis infections. This disorder has rarely been diagnosed in the Spanish population. In this work we report the immunochemical and molecular characterization of complement C6 deficiency in a Spanish patient showing no detectable functional activity of either the classical or alternative complement pathways and reporting a history of several episodes of meningococcal meningitis. The levels of individual complement components C3, C4, C5, C7, C8 and C9 were within the normal range. However, C6 level was low in the patient's serum as measured by radial immunodiffusion. Exon-specific polymerase chain reaction and sequencing of the C6 gene revealed a previously described homozygous single base deletion in exon 6 (c.821delA), leading to a shift in the reading frame that caused the generation of a downstream stop codon, which, in turn, provoked the truncation of the C6 protein (p.Gln274fs). To our knowledge, this is the first report on the c.821delA mutation in the Spanish population, which has previously only been identified in individuals of African ancestry. Characterization of this mutation was thought interesting in order to elucidate its source and help understand the molecular basis of this uncommon deficiency in our population. Moreover, this report highlights the importance of complement screening in cases of repeated meningococcal infections in order to establish its involvement and to consider adequate clinical recommendations such as prophylactic antibiotics or meningococcal vaccines and, subsequently, for genetic counselling.

Seasonal input of regulated and emerging organic pollutants through surface watercourses to a Mediterranean coastal lagoon.

Seasonal input of organic pollutants through El Albujón Watercourse to the Mar Menor lagoon was estimated from Spring 2009 to Winter 2010, including regular periods and two flash flood events. 82 semivolatile organic pollutants (persistent organic pollutants, different groups of pesticides and others) were determined by stir bar sorptive extraction and thermal desorption followed by capillary gas chromatography coupled to mass spectrometry from surface waters with quantification limits of a few ngL(-1). Pesticide concentrations varied significantly along the watercourse due to the presence of different sources (groundwaters, wastewater effluent, tributary contributions, brackish waters, etc.) and physicochemical/biological processes that take place simultaneously. The most commonly detected analytes were propyzamide, triazine compounds and chlorpyrifos. A clear seasonal pattern has been detected, with a predominance of insecticides during Summer and of herbicides during Winter. The input of pesticides through this watercourse is particularly relevant during periods of heavy rain, representing more than 70% of total yearly input for many of them.

Spatial distribution and temporal trends of polycyclic aromatic hydrocarbons in Mytilus galloprovincialis from the Iberian Mediterranean coast.

Polycyclic aromatic hydrocarbons (PAHs) have been determined in blue mussels (Mytilus galloprovincialis) from several Iberian Mediterranean coastal areas through the implementation of a monitoring programme from Spain in the framework of the Mediterranean Pollution Programme (MED POL). The selected areas correspond to sites with differing degrees of exposure to the main pollution sources (hot spots, coastal and reference areas). The sampling campaigns were performed from 2004 to 2009, with samples being taken from May to June, the non-spawning period for mussels in this area. Thirteen PAHs were determined by high-performance liquid chromatography with specific fluorescence detection. In general, total PAHs concentration was lower than 50 µg kg(-1) d.w., except in areas close to the principal ports and cities (Barcelona, Tarragona, Valencia and Algeciras) where it varies from 75 to 390 µg kg(-1) d.w. Background concentrations have been proposed for PAHs in mussels (23.8 µg kg(-1) d.w.) from Western Mediterranean area. Temporal trends were not statistically significant for PAHs concentrations from 2004 to 2009. Longer monitoring periods would be required to detect a continuous tendency, especially for PAHs because although the efficiency of combustion engines has reduced PAHs emissions, their increasing use could alter this potential reduction. The predominant PAHs were three and four ring congeners in all cases, with the predominance of phenanthrene in mussels sited far from the main PAHs sources. The phenanthrene/anthracene (lower than 10) and fluoranthene/pyrene (higher than 1) ratios indicate that PAHs detected in Spanish Mediterranean coastal mussels are mainly of pyrolytic origin.

Post-transplant increase in soluble human leukocyte antigen-G associated with non-severe cardiac allograft vasculopathy.

Cardiac allograft vasculopathy (CAV) is the single most important long-term limitation to heart transplantation. This study aimed to assess the value of monitoring soluble human leukocyte antigen-G (sHLA-G) during the first year post-transplantation to predict the severity of CAV, in 21 out of 77 heart recipients assessed by intravascular ultrasound (IVUS). Serum sHLA-G concentration increased after transplant in recipients free of severe CAV, but decreased in recipients suffering from severe CAV, significant differences between these two groups were found 6 to 12 months post-transplantation. The optimal value of the change in post-transplant sHLA-G for identifying severe CAV was ≥0.062%, which maximized sensitivity (80%) and specificity (100%). Importantly, increases in post-transplant sHLA-G were inversely associated with severe CAV, but directly associated with human cytomegalovirus reactivation. In addition, recipients presenting non-severe CAV or an increased sHLA-G post-transplantation, showed higher numbers of CD8(+)CD28(-) T cells and a down-modulation of CD28 on CD4(+) lymphocytes, which typically identifies CD8(+) regulatory T cells and anergic/tolerogenic T helper cells, respectively. In conclusion, quantification of sHLA-G might offer a complementary non-invasive method for identifying recipients at risk of more severe CAV and who might benefit from earlier preventive therapies, although these results need to be confirmed in larger series.

C1q-fixing human leukocyte antigen assay in immunized renal patients: correlation between Luminex SAB-C1q and SAB-IgG.

BACKGROUND: There is no consensus about the impact of thresholds of complement-fixing antibody assays. Recently, a C1q-SAB assay has been developed to identify complement-fixing HLA antibodies with high sensitivity and specificity. Our aim was to determine the correlation between IgG single antigens beads (SAB) and C1q-SAB assay results among patients on the renal waiting list. PATIENTS AND METHODS: Serum samples from immunized renal waiting list patients as well as negative and positive controls were valided by Luminex (LMX). These sera, which were positive for 166 antibody specificities, were tested for HLA class I in parallel by LMX-IgG and LMX-C1q. RESULTS: Comparison of antibody detection revealed no correlation based on median fluorescent intensity (MFI), levels between the IgG SAB and the C1qSAB assay (P > .05). IgG-positive sera with MFIs as low as 700 were able to fix C1q, whereas other sera with MFIs as high 14,500 did not. Furthermore, there appeared to be disparities in the profiles of class I antigens able to fix C1q-SAB. In our series, only 34% class I IgG SAB antibodies were also C1qSAB+. In several patients, we detected C1qSAB+ against IgGSAB- that was surely due to IgM antibodies. So, the C1qSAB assay detected IgM antibodies that fix complement. CONCLUSION: These data suggested that the C1q-SAB assay could be an important method to evaluate pretransplant virtual crossmatch and to define nonpermitted specificities (C1q-fixing) in kidney transplantation.

Health status of red mullets from polluted areas of the Spanish Mediterranean coast, with special reference to Portmán (SE Spain).

A suite of general physiological indicators and hepatic biomarker responses were determined in red mullet (Mullus barbatus) from priority pollution areas of W Mediterranean Sea, including the highly metal polluted area of Portmán (Cartagena, SE Spain). Concentrations of metals and persistent organic pollutants in fish muscle tissue and sediment samples were also analysed. Our results showed that fish from Portmán accumulated the highest concentrations of mercury, lead and arsenic and also of some polycyclic aromatic hydrocarbons and polychlorinated biphenyls congeners. In addition, they had significantly lower condition factor, muscle lipid content and gonadosomatic index, as well as the lowest levels of DNA integrity and the highest ethoxyresorufin-O-deethylase (EROD) activity in liver of the areas investigated. Contaminant body burden in fish only partly corresponded to chemical characteristics of the sediments in the areas in which they were collected. Our findings indicate that red mullets from Portmán had suboptimal health status that warrant further study.

CD28 and KIR2D receptors as sensors of the immune status in heart and liver transplantation.

Viral infections and cellular acute rejection (AR) condition immunosuppressive therapy and compromise the evolution of allografts. Immune monitoring can be useful for ascertaining rejection and for differentiating allo-reaction from activation induced by infections. This work analyzes the usefulness of monitoring the expression of CD28 and KIR2D receptors in peripheral blood T lymphocytes by flow cytometry, to ascertain the immune response in heart and liver transplant recipients. In both types of transplant, the up-regulation of CD28 in CD4(+) lymphocytes in the periods of greatest AR frequency indicates an effective allo-response, whereas the post-transplantation emergence of circulating CD8(+)CD28(-) and CD8(+)CD28(-)KIR2D(+) T cells correlates with better early clinical results. Cytomegalovirus (CMV) infection, but not hepatitis C virus (HCV) or other infections, abrogated both CD28 up-regulation and CD8(+)CD28(-)KIR2D(+) T-cell expansion. Our results show that monitoring the expression of CD28 and KIR2D receptors on T lymphocytes might be considered as sensors of the immune status of heart and liver recipients.

Divergences in KIR2D+ natural killer and KIR2D+CD8+ T-cell reconstitution following liver transplantation.

Natural killer (NK) and CD8(+) T cells may be active elements in the allograft response, but little is known about their role in liver transplantation. Some of these cells express killer immunoglobulin-like receptors (KIRs), which after binding specific ligands may transmit inhibitory/activating signals. In this study, circulating NK and CD8(+) T cells expressing CD158a/h (KIR2DL1/S1) or CD158b/j (KIR2DL2/3/S(2)) receptors were analyzed in 142 liver recipients by flow cytometry. They were underrepresented in patients before transplantation, but following transplantation, whereas the KIR2D(+) NK subsets experienced a late recuperation (day 365) mainly in C2-homozygous patients developing early acute rejection, recovery of the 2 CD8(+)KIR2D(+) T cells started earlier, showing significant differences on day 365 between patients without acute rejection and those suffering from it (p = 0.004 and p < 0.0001, respectively). These differences were also evident when the human leukocute antigen-C genotypes of the recipient were considered. In conclusion, whereas the late recovery of KIR2D(+) NK cells in C2/C2 patients appears to be linked to acute rejection, the increase in early CD8(+)KIR2D(+) T cells in overall liver recipients correlates with a most successful early graft outcome. Therefore, monitoring of KIR2D(+) cells appears to be a useful tool for liver transplant follow-up.

Antioxidant responses in gills of mussel (Mytilus galloprovincialis) as biomarkers of environmental stress along the Spanish Mediterranean coast.

Antioxidant response was used to assess the effects of the main pollutants in wild mussels (Mytilus galloprovincialis) along the Mediterranean coast of Spain. Antioxidant enzyme activities - those of catalase, superoxide dismutase, glutathione peroxidases, glutathione reductase, glutathione S-transferase and DT-diaphorase - as well as lipid peroxidation and metallothionein concentrations were measured in gills of mussels from 16 selected sites. Furthermore, concentrations of the main contaminants (Hg, Pb, Cd, Cu, Zn, As, PAH, PCB, and DDT) were quantified in mussel tissue, and environmental parameters were measured in water samples collected at each site. Results showed that the glutathione-dependent antioxidant enzymes offered an increased and coordinated response against metal (Hg, Pb and Cd) contamination. These enzymatic activities correlated positively to temperature, suggesting the influence of this environmental parameter on antioxidant responses in gill tissues. Furthermore, although temperature did not reach stressful levels in the study area, it seemed to add a synergistic effect to that produced by metals to induce antioxidant enzymes in the most metal-polluted sites. Catalase activity appeared to be involved in a different antioxidant pathway, more related to organic pollutant bioaccumulation, offering an efficient protection mechanism against reactive oxygen species generation due both to organic exposure and high physiological activity, reflected by high condition indices. In general terms, increased levels of antioxidant enzymes at some sites suffering from metal and organic pollution indicated a situation of oxidative stress that nevertheless did not appear to be harmful, since lipid peroxidation levels showed no peroxidative damage in gill tissues of mussels collected from even the most heavily polluted sites. On the other hand, metallothionein and DT-diaphorase did not reflect pollutant exposure and seemed to be more influenced by environmental variables than by the pollutants.