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The biomonitoring of metallic contamination in marine ecosystems is often focused on animal species of commercial interest and in lesser extent on non-model marine invertebrates. The aim of this study was to compare the metal concentrations (Li, Al, Ti, Cr, Fe, Ni, Cu, Zn, As, Ag, Cd, Hg, Pb) in seven marine sponges with a particular interest in the homoscleromorph sponge Oscarella lobularis at different sites of the Bay of Marseille, France. Inter-species variabilities suggest that the seven sponge species studied accumulate metals differently. In O. lobularis , a multi-site analysis shows different bioaccumulation between the eight sampled populations. These inter-site differences may reflect differences in the hydrodynamic features and in past and present industrial activities. Because Oscarella lobularis shows a homogeneous metal accumulation pattern in comparison with the other tested species, it appears to be suitable for metal contamination biomonitoring in Mediterranean coastal waters, in particular of the coralligenous communities. [Display omitted] • The sponge Oscarella lobularis shows site-dependent metal concentrations. • Metals are differently accumulated between sponge species at the same site. • O. lobularis can be a suitable bioindicator of metallic contamination of coastal waters. [ABSTRACT FROM AUTHOR]

Introduction Like other tissues in the body, the human dental pulp is equipped with a network of immune cells that can be mobilized against pathogens when they invade the tooth. Very little data, mostly obtained with classic histologic methods, have reported their quantities and relative percentages. The objective of this study was to characterize and precisely quantify immunocompetent cells in healthy human dental pulp by using fluorescence-activated cell sorting, together with identifying specific cell subsets in the leukocyte (CD45 + ) cells. Methods Healthy human third molars were collected from 42 young patients. Dental pulps were separated from the hard tissues and prepared for flow cytometry or immunostaining analyses. Results CD45 + cells represented 0.94% ± 0.65% of cells obtained from the enzymatic digestion of whole dental pulps ( n = 34). CD16 + CD14 + granulocytes/neutrophils (50.01% ± 9.08%, n = 7) were found to represent the major subpopulation in CD45 + cells followed by CD3 + T lymphocytes (32.58% ± 11%, n = 17), CD14 + monocytes (8.93% ± 5.8%, n = 7), and HLA-DR high Lin1 - dendritic cells (4.51% ± 1.12%, n = 7). Minor subpopulations included CD3 − CD56 + natural killer cells (2.63% ± 1.15%, n = 7) and CD19 + B lymphocytes (1.65% ± 0.89%, n = 17). We further identified cells harboring a phenotype compatible with Foxp3/CD25-expressing regulatory T lymphocytes (CD45 + CD3 + CD4 + CD127 low ). Fluorescence-activated cell sorting analysis and confocal microscopy also revealed expression of HO-1 in HLA-DR + cells. Conclusions For the first time, this study identifies and precisely quantifies the relative proportion of immunocompetent cells potentially involved in tissue homeostasis of healthy human dental pulp. [ABSTRACT FROM AUTHOR]

Highlights: [•] VAR2CSA ID1-DBL2Xb is the leading candidate for the development of a vaccine against pregnancy-associated malaria. [•] It displays substantial sequence polymorphism in natural P. falciparum populations. [•] Most sequence variants are common and extensively shared among worldwide parasite populations. [•] Several peaks of strong population differentiation are observed at specific polymorphic loci. [Copyright &y& Elsevier]

Highlights: [•] Plasmodium falciparum gene polymorphisms of mothers’ and infants’ infections were compared. [•] Geographic Information System concluded on the random repartition of glurp alleles. [•] In infants, infections with shared placental-infant glurp alleles were favored. [•] Plasmodial antigen polymorphism contributes to the structuring of specific immunity. [ABSTRACT FROM AUTHOR]

Mediterranean marine biota suffers from various anthropogenic threats. Among them, pollutants such as mercury (Hg) represent important environmental issues that are exacerbated by bioaccumulation and bioamplification along food webs via its organic form, monomethylmercury (MMHg). To date, very little is known regarding the impact of mercury on Porifera and the few available studies have been exclusively focused on Demospongiae. This work studies the effect of MMHgCl at different biological levels of Oscarella lobularis (Porifera, Homoscleromorpha). Bioaccumulation assays show that MMHgCl significantly accumulated in sponge tissues after a 96-h exposure to 0.1 μg L−1. Toxicity assays (LC50 96h) show a sensibility that depends on life-stage (adult vs bud). Additionally, we show that the exposure to 1 μg L−1 MMHgCl negatively impacts the epithelial integrity and the regeneration process in buds, as shown by the loss of cell-cell contacts and the alteration of osculum morphogenesis. For the first time in a sponge, a whole set of genes classically involved in metal detoxification and in antioxidant response were identified. Significant changes in catalase, superoxide dismutase and nuclear factor (erythroid-derived 2)-like 2 expressions in exposed juveniles were measured. Such an integrative approach from the physiological to the molecular scales on a non-model organism expands our knowledge concerning sensitivity and toxicity mechanisms induced by MMHg in Porifera, raising new questions regarding the possible defences used by marine sponges. [Display omitted] • The sensitivity of the sponge Oscarella lobularis life cycle-dependent. • Three antioxidant genes are under-expressed after exposure to mercury. • Exposure to mercury induces strong defects on major epithelial features. [ABSTRACT FROM AUTHOR]

Transcriptional mechanisms regulating type I collagen genes expression in physiopathological situations are not completely known. In this study, we have investigated the role of nuclear factor-κB (NF-κB) transcription factor on type I collagen expression in adult normal human (ANF) and scleroderma (SF) fibroblasts. We demonstrated that NF-κB, a master transcription factor playing a major role in immune response/apoptosis, down-regulates COL1A1 expression by a transcriptional control involving the -112/-61 bp sequence. This 51-bp region mediates the action of two zinc fingers, Sp1 (specific protein-1) and Sp3, acting as trans-activators of type I collagen expression in ANF and SF. Knockdown of each one of these trans factors by siRNA confirmed the trans-activating effect of Sp1/Sp3 and the p65 subunit of NF-κB trans-inhibiting effect on COL1A1 expression. Despite no existing κB consensus sequence in the COL1A1 promoter, we found that Sp1/Sp3/c-Krox and NF-κB bind and/or are recruited on the proximal promoter in chromatin immunoprecipitation (ChIP) assays. Attempts to elucidate whether interactions between Sp1/Sp3/c-Krox and p65 are necessary to mediate the NF-κB inhibitory effect on COL1A1 in ANF and SF were carried out; in this regard, immunoprecipitation assays revealed that they interact, and this was validated by re-ChIP. Finally, the knockdown of Sp1/Sp3/c-Krox prevents the p65 inhibitory effect on COL1A1 transcription in ANF, whereas only the siRNAs targeting Sp3 and c-Krox provoked the same effect in SF, suggesting that particular interactions are characteristic of the scleroderma phenotype. In conclusion, our findings highlight a new mechanism for COL1A1 transcriptional regulation by NF-κB, and these data could allow the development of new antifibrotic strategies. [ABSTRACT FROM AUTHOR]

Type II collagen is composed of α1(II) chains encoded by the COL2A1 gene. Alteration of this cartilage marker is a common feature of osteoarthritis. Interleukin-6 (IL-6) is a pro-inflammatory cytokine that needs a soluble form of receptor called sIL-6R to exert its effects in some cellular models. In that case, sIL-6R exerts agonistic action. This mechanism can make up for the partial or total absence of membrane-anchored IL-6 receptors in some cell types, such as chondrocytes. Our study shows that IL-6, sIL-6R, or both inhibit type II collagen production by rabbit articular chondrocytes through a transcriptional control. The cytokine and/or sIL-6R repress COL2A1 transcription by a -63/-35 sequence that binds Sp1·Sp3. Indeed, IL-6 and/or sIL-6R inhibit Spi and Sp3 expression and their binding activity to the 63-bp promoter. In chromatin immunoprecipitation experiments, IL-6·sIL-6R induced an increase in Sp3 recruitment to the detriment of Spi. Knockdown of Sp1·Sp3 by small interference RNA and decoy strategies were found to prevent the IL-6- and/or sIL-6R-induced inhibition of COL2A1 transcription, indicating that each of these Sp proteins is required for down-regulation of the target gene and that a heterotypic Sp1·Sp3 complex is involved. Additionally, Spi was shown to interact with Sp3 and HDAC1. Indeed, overexpression of a full- length Sp3 cDNA blocked the Spi up-regulation of the 63-bp COL2A1 promoter activity, and by itself, inhibits COL2A1 transcription. We can conclude that IL-6, sIL-6R, or both in combination decrease both the Sp1·Sp3 ratio and DNA-binding activities, thus inhibiting COL2A1 transcription. [ABSTRACT FROM AUTHOR]

Despite several investigations, the transcriptional mechanisms that regulate the expression of both type I collagen genes (COL1A1 and COL1A2) in either physiological or pathological situations, such as scleroderma, are not completely known. We have investigated the role of hc-Krox transcription factor on type I collagen expression by human dermal fibroblasts. hc-Krox exerted a stimulating effect on type I collagen protein synthesis and enhanced the corresponding mRNA steady-state levels of COL1A1 and COL1A2 in foreskin fibroblasts (FF), adult normal fibroblasts (ANF), and scleroderma fibroblasts (SF). Forced hc-Krox expression was found to up-regulate COL1A1 transcription through a –112/–61-bp sequence in FF, ANF, and SF. Knockdown of hc-Krox by short interfering RNA and decoy strategies confirmed the transactivating effect of hc-Krox and decreased substantially COL1A1 transcription levels in all fibroblast types. The –112/–61-bp sequence bound specifically hc-Krox but also Sp1 and CBF. Attempts to elucidate the potential interactions between hc-Krox, Sp1, and Sp3 revealed that all of them co-immunoprecipitate from FF cellular extracts when a c-Krox antibody was used and bind to the COL1A1 promoter in chromatin immunoprecipitation assays. Moreover, hc-Krox DNA binding activity to its COL1A1-responsive element is increased in SF, cells producing higher amounts of type I collagen compared with ANF and FF. These data suggest that the regulation of COL1A1 gene transcription in human dermal fibroblasts involves a complex machinery that implicates at least three transcription proteins, hc-Krox, Sp1, and Sp3, which could act in concert to up-regulate COL1A1 transcriptional activity and provide evidence for a pro-fibrotic role of hc-Krox. [ABSTRACT FROM AUTHOR]

Members of chitinase-like proteins (CLPs) have attracted much attention because of their ability to promote cell proliferation in insects (imaginal disc growth factor) and mammals (YKL-40). To gain insights into the molecular processes underlying the physiological control of growth and development in Lophotrochozoa, we report here the cloning and biochemical characterization of the first Lophotrochozoan CLP from the oyster Crassostrea gigas (Cg-CIp1). Gene expression profiles monitored by real time quantitative reverse transcription-PCR in different adult tissues and during development support the involvement of this protein in the control of growth and development in C. gigas. Recombinant Cg-CIp1 demonstrates a strong affinity for chitin but no chitinolytic activity, as was described for the HC-gp39 mammalian homolog. Furthermore, transient expression of Cg-CIp1 in primary cultures of rabbit articular chondrocytes as well as the use of both purified recombinant protein and conditioned medium from Cg-CIp1-expressing rabbit articular chondrocytes established that Cg-CIp1 stimulates cell proliferation and regulates extracellular matrix component synthesis, showing for the first time a possible involvement ot a CLP on type II collagen synthesis regulation. These observations together with the fact that Cg-CIp1 gene organization strongly resembles that of its mammalian homologues argue for an early evolutionary origin and a high conservation of this class of proteins at both the structural and functional levels. [ABSTRACT FROM AUTHOR]

Interleukin-1β (IL-1β) is a pleiotropic cytokine that was shown to inhibit the biosynthesis of articular cartilage components. Here we demonstrate that IL-1β inhibits the production of newly synthesized collagens in proliferating rabbit articular chondrocytes and that this effect is accompanied by a decrease in the steady-state levels of type II collagen mRNA. IL-1β down-regulates COL2A1 gene transcription through a -41/-33 bp sequence that binds a multimeric complex including Sp1 and Sp3 transcription factors. Specificity of IL-1β effects on COL2A1 promoter activity was demonstrated in experiments in which transfection of a wild type -50/+1 sequence of COL2A1 promoter as a decoy oligonucleotide abolished the IL-1β inhibition of a -63/+47 COL2A1-mediated transcription. By contrast, transfection of the related oligonucleotide harboring a targeted mutation in the -41/-33 sequence did not modify the negative effect the cytokine. Because we demonstrated previously that Sp1 was a strong activator of COL2A1 gene expression via the -63/+1 promoter region, whereas Sp3 overexpression blocked Sp1-induced promoter activity and inhibited COL2A1 gene transcription, we conclude that IL-1β down-regulation of that gene, as we found previously for transforming growth factor-β1, is mediated by an increase in the Sp3/Sp1 ratio. Moreover, IL-1β increased steady-state levels of Sp1 and Sp3 mRNAs, whereas it enhanced Sp3 protein expression and inhibited Sp1 protein biosynthesis. Nevertheless, IL-1β decreased the binding activity of both Sp1 and Sp3 to the 63-bp short COL2A1 promoter, suggesting that the cytokine exerts a post-transcriptional regulatory mechanism on Sp1 and Sp3 gene expressions. Altogether, these data indicate that modulation of Sp3/Sp1 ratio in cartilage could be a potential target to prevent or limit the tissue degradation. [ABSTRACT FROM AUTHOR]