LOH at 6p21.3 region and HLA class altered phenotypes in bladder carcinomas

Alterations in HLA class I antigen expression have been frequently described in different epithelial tumors and are thought to favor tumor immune escape from T lymphocyte recognition. Multiple molecular mechanisms are responsible for these altered HLA class I tumor phenotypes. Some are structural defects that produce unresponsiveness to treatment with interferons. Others include alterations in regulatory mechanisms that can be switched on by treatment of tumor cells with different cytokines. One important mechanism belonging to the first group is loss of heterozygosity (LOH) at chromosome region 6p21.3, which can lead to HLA haplotype loss. In this investigation, the frequency of LOH at 6p21 chromosome region was studied in 69 bladder carcinomas. Short tandem repeat analysis showed that 35% of cases had LOH in this chromosome region. By considering these results together with immunohistological findings previously published by our group, we identified a distribution pattern of HLA class I altered phenotypes in bladder cancer. The most frequently altered phenotype in bladder carcinomas was total loss of HLA class I expression (17 cases, 25%), followed by phenotype II associated with HLA haplotype loss (12 cases, 17.5%), and HLA allelic loss (ten cases, 14.5%). Nine cases (13%) were classified as having a compound phenotype, five cases (7%) as having HLA locus loss, and in 16 cases (23%) no alteration in HLA expression was detected. An important conclusion of this report is that a combination of different molecular and immunohistological techniques is required to precisely define which HLA alleles are lost during tumor progression and to characterize the underlying mechanisms of these losses. These studies should be performed when a cancer patient is to be included in an immunotherapy protocol that aims to stimulate different immune effector mechanisms.     

Estrogen protects renal endothelial barrier function from ischemia-reperfusion in vitro and in vivo

Emerging evidence suggests that renal endothelial function may be altered in ischemia-reperfusion injury. Acute kidney injury is sexually dimorphic, and estrogen protects renal tubular function after experimental ischemic injury. This study tested the hypothesis that during ischemia-reperfusion, estrogen alters glomerular endothelial function to prevent hyperpermeability. Glomerular endothelial cells were exposed to 8-h oxygen-glucose deprivation (OGD) followed by 4- and 8-h reoxygenation-glucose repletion. After 4-h reoxygenation-glucose repletion, transendothelial permeability to Ficoll-70 was reduced, and transendothelial resistance increased, by 17β-estradiol vs. vehicle treatment during OGD (OGD-vehicle: 91.0 ± 11.8%, OGD-estrogen: 102.6 ± 10.8%, P < 0.05). This effect was reversed by coadministration of G protein-coupled receptor 30 (GPR30) antagonist G15 with 17β-estradiol (OGD-estrogen-G15: 89.5 ± 6.9, P < 0.05 compared with 17β-estradiol). To provide preliminary confirmation of this result in vivo, Ficoll-70 was administered to mice 24 h after cardiac arrest and cardiopulmonary resuscitation (CA/CPR). Blood urea nitrogen (BUN) and serum creatinine (SCr) in these mice were elevated within 12 h following CA/CPR and reduced at 24 h by pretreatment with 17β-estradiol (BUN/SCr 17β-estradiol: 34 ± 19/0.2 ± 0.1 vehicle: 92 ± 49/0.5 ± 0.3, n = 8-12, P < 0.05). Glomerular sieving of Ficoll 70 was increased by CA/CPR within 2 h of injury and 17β-estradiol treatment (θ; 17β-estradiol: 0.74 ± 0.26 vs. vehicle: 1.05 ± 0.53, n = 14-15, P < 0.05). These results suggest that estrogen reduces postischemic glomerular endothelial hyperpermeability at least in part through GPR30 and that estrogen may regulate post CA/CPR glomerular permeability in a similar fashion in vivo.     

Assessing Dispersal Patterns of Fish Propagules from an Effective Mediterranean Marine Protected Area

Successfully enforced marine protected areas (MPAs) have been widely demonstrated to allow, within their boundaries, the recovery of exploited species and beyond their boundaries, the spillover of juvenile and adult fish. Little evidence is available about the so-called ‘recruitment subsidy’, the augmented production of propagules (i.e. eggs and larvae) due to the increased abundance of large-sized spawners hosted within effective MPAs. Once emitted, propagules can be locally retained and/or exported elsewhere. Patterns of propagule retention and/or export from MPAs have been little investigated, especially in the Mediterranean. This study investigated the potential for propagule production and retention/export from a Mediterranean MPA (Torre Guaceto, SW Adriatic Sea) using the white sea bream, Diplodus sargus sargus, as a model species. A multidisciplinary approach was used combining 1) spatial distribution patterns of individuals (post-settlers and adults) assessed through visual census within Torre Guaceto MPA and in northern and southern unprotected areas, 2) Lagrangian simulations of dispersal based on an oceanographic model of the region and data on early life-history traits of the species (spawning date, pelagic larval duration) and 3) a preliminary genetic study using microsatellite loci. Results show that the MPA hosts higher densities of larger-sized spawners than outside areas, potentially guaranteeing higher propagule production. Model simulations and field observation suggest that larval retention within and long-distance dispersal across MPA boundaries allow the replenishment of the MPA and of exploited populations up to 100 km down-current (southward) from the MPA. This pattern partially agrees with the high genetic homogeneity found in the entire study area (no differences in genetic composition and diversity indices), suggesting a high gene flow. By contributing to a better understanding of propagule dispersal patterns, these findings provide crucial information for the design of MPAs and MPA networks effective to replenish fish stocks and enhance fisheries in unprotected areas.     

Investigation of Ag in the king scallop Pecten maximus using field and laboratory approaches

The bioaccumulation, tissue and subcellular distributions of Ag were investigated in the king scallop Pecten maximus from the Bay of Seine fishery area (France) in laboratory and in field conditions. Experimental investigations with the radiotracer ^110mAg showed that the scallop readily concentrated Ag when exposed via seawater and to a much lower extent when exposed via sediment. Retention of the metal incorporated via all tested contamination pathways was shown to be very strong, but the assimilation efficiency of Ag ingested with food was found to be tightly depending on the phytoplankton strain used to feed the scallops (74 and 33% with Skeletonema costatum and Isochrysis galbana, respectively). The uptake and depuration kinetic parameters determined in the laboratory experiments were used to run a global bioaccumulation model. The latter indicated that the major uptake pathway of Ag in P. maximus was strongly depending on the food quality. Indeed, when fed the diatom S. costatum which was characterised by a high affinity for Ag (high distribution constant K_df) the relative contribution of the feeding pathway reached 98% of the global Ag bioaccumulation. In contrast, when fed I. galbana which displayed a lower K_df than S. costatum, dietary Ag was retained to a lesser extent by P. maximus, and seawater appeared as the major contributing uptake pathway. In wild scallops collected from reference and contaminated sites, Ag was mainly concentrated in the digestive gland and secondarily in the gills, and was mainly found associated with the insoluble subcellular fraction in all the scallop tissues.     

Habituation of wild chimpanzees (Pan troglodytes) of the South Group at Taï Forest, Côte d'Ivoire: empirical measure of progress

The last part of the chimpanzee (Pan troglodytes) habituation process of the Ta? South Group was monitored over 2 years (1994-1996), during which all males and most of the females became habituated to observation by humans. The time needed (5-7 years) to habituate the community was similar to that recorded for the Ta? North Group and is comparable to other populations studied in Africa. Variation emerged in habituation rate: males were habituated earlier than females, and among females, sexually cycling individuals were habituated faster than non-cycling females. Such differences may be a function of both the methods used to find the chimpanzees and the sex of the individual. Reproductive status and individuality may also have influenced habituation rates by affecting the number of contacts required to habituate a chimpanzee to neutral humans.     

The potential role of spherocrystals in the detoxification of essential trace metals following exposure to Cu and Zn in the fighting conch <Emphasis Type="Italic">Strombus</Emphasis> (<Emphasis Type="Italic">Lobatus</Emphasis>) <Emphasis Type="Italic">pugilis</Emphasis>

Crypt cells—one of the three cell types composing Strombidae digestive tubules—are characterized by the presence of numerous metal-containing phosphate granules termed spherocrystals. We explored the bioaccumulation and detoxification of metals in Strombidae by exposing wild fighting conch Strombus pugilis for 9 days to waterborne CuSO₄ and ZnSO₄. The total amount of Cu and Zn was determined in the digestive gland and in the rest of the body by Inductively Coupled Plasma (ICP) analyses. The digestive gland spherocrystal metal content was investigated based on the semi-quantitative energy dispersive X-ray (EDX) elemental analysis. ICP analyses of unexposed individuals revealed that 87.0?±?5.9% of the Zn is contained in the digestive gland, where its concentration is 36 times higher than in the rest of the body. Regarding Cu, 25.8?±?16.4% of the metal was located in the digestive gland of the control individuals, increasing to 61.5?±?16.4% in exposed individuals. Both Cu and Zn concentrations in the digestive gland increased after exposures, pointing to a potential role of this organ in the detoxification of these metals. EDX analysis of spherocrystals revealed the presence of Ca, Cl, Fe, K, Mg, P, and Zn in unexposed individuals. No difference was found in the relative proportion of Zn in spherocrystals of exposed versus control individuals. Contrastingly, copper was never detected in the spherocrystals from controls and Zn-exposed individuals, but the relative proportion of Cu in spherocrystals of Cu-exposed individuals varied from 0.3 to 5.7%. Our results show the direct role of spherocrystals in Cu detoxification.     

A mass stranding of seven Longman's beaked whales (Indopacetus pacificus) in New Caledonia,South Pacific

Seven Longman's beaked whales (Indopacetus pacificus) stranded together in southern New Caledonia on 16 November 2013 (one adult male, two adult females, two subadult females, one calf, and one unknown). At this time, we have no evidence to suggest that this event was an “atypical” mass stranding associated with active naval sonar or other anthropogenic activities. The adult females were slightly larger (618–640 cm) than the adult male (590 cm). The length of the calf (ca. 300 cm) suggests it was less than a year old. Five of the whales were sampled for mitochondrial (mt) DNA analysis to confirm species identification. All shared the same haplotype over 680 bp of the mtDNA control region. High concentrations of Hg, Fe, Se, Zn (all in the liver), and Cd (in the kidneys) were detected. Necropsies revealed plastic debris in the stomach of two of the whales. One of these same whales had chronic gastritis while the other had acute pleurisy and also tested positive for morbillivirus. This infection may have been a major factor behind this mass stranding event.     

No longer Demospongiae: Homoscleromorpha formal nomination as a fourth class of Porifera

Over the past few years, there has been growing interest among the sponge community in the phylogenetic position of the Homoscleromorpha (i.e. within or outside the class Demospongiae). Recent molecular analyses clearly show that the Homoscleromorpha forms a distinct clade separated from the Demospongiae and is composed of two families, Oscarellidae and Plakinidae. Within the currently more widely accepted hypothesis of a monophyletic Porifera, we formally propose here to raise Homoscleromorpha to the class rank (the fourth one). We, therefore, provide a definition and a formal diagnosis. In the supplementary materials, we also present an alternative classification of the Homoscleromorpha, following the PhyloCode.     

Accounting for transportation impacts in the environmental assessment of waste management plans

Background, aim, and scope  Many recent studies on waste management have described in detail the potential impacts of recycling and final treatment of municipal waste. In public debates, the attention has also been focused on the choice of final disposal technologies (e.g. landfilling vs. incineration). However, a comprehensive assessment of the impacts of waste collection and transport was still lacking. In the present study, we use LCA to evaluate the potential impact of the provincial waste management plan of Varese (northern Italy). Particular attention is devoted to the estimation of environmental impacts generated during waste transport. Materials and methods  A detailed Life Cycle Inventory was built for the transportation phase, based on primary data collected by interviewing the agencies involved in waste collection. To model the recycling and final disposal phase we relied on the BUWAL 250 database. Impacts were evaluated with the Eco-Indicator 99 method in its egalitarian formulation. Results  The results of our analysis reveal that the major potential impacts of the plan are associated with waste collection and transport. These impacts are partially compensated by reduced resource consumption through recycling and energy recovery through incineration. Discussion  The outputs of the LCIA were compared with those obtained by using other ecoindicators (Eco-Indicator 99 hierarchist and individualist, CML2, EPS2000). Although not comparable on a quantitative basis, they are qualitatively consistent. Conclusions  Neglecting the effects of collection and transport might result in a severe underestimation of the environmental impacts of a waste management system, especially as refers to depletion of fossil fuels, emission of respiratory inorganics and climate change. To reduce the environmental impact of waste management systems, an accurate optimisation of waste transport is required. Recommendations and perspectives  Effective waste management planning requires the explicit inclusion of waste collection and transport when comparing alternative management policies.     

Gender-associated differences of perforin polymorphisms in the susceptibility to multiple sclerosis

The granule-dependent exocytosis pathway is an important mechanism to induce apoptosis by CD8(+) T cells and NK cells and involves lytic molecules such as perforin. In the current study, we investigated the perforin 1 gene (PRF1) as a candidate for multiple sclerosis (MS) susceptibility in the Spanish population. We genotyped three PRF1 single nucleotide polymorphisms (rs885822, rs10999426, and rs3758562) in 420 patients with MS and 512 controls. Associations of PRF1 polymorphisms with the disease were restricted to male patients with MS, and the finding was consistently observed at the allele, genotype, and haplotype levels. Gender-associated differences were validated in an additional replication cohort comprised of 292 MS cases and 300 controls. In addition, we identified minor risk haplotypes strongly associated with male patients having primary progressive MS (PPMS). Further characterization of male patients with PPMS carrying the risk haplotypes by means of gene expression microarrays revealed overrepresentation of the cell killing gene ontology category among downregulated genes in these patients compared with male patients with PPMS carrying protective haplotypes. Moreover, PRF1 mRNA expression levels were significantly lower in patients with risk haplotypes, and changes in perforin protein expression by CD8(+) T cells mirrored those observed in gene expression. These findings suggest a gender dimorphism in the PRF1 association with MS and point to the presence of a generalized defect in the expression of genes that code for proteins involved in cell killing in a subgroup of male patients with PPMS.