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1.
Sci Rep ; 9(1): 3718, 2019 03 06.
Article in English | MEDLINE | ID: mdl-30842491

ABSTRACT

The biogeochemistry of hypersaline environments is strongly influenced by changes in biological processes and physicochemical parameters. Although massive evaporation events have occurred repeatedly throughout Earth history, their biogeochemical cycles and global impact remain poorly understood. Here, we provide the first nitrogen isotopic data for nutrients and chloropigments from modern shallow hypersaline environments (solar salterns, Trapani, Italy) and apply the obtained insights to δ15N signatures of the Messinian salinity crisis (MSC) in the late Miocene. Concentrations and δ15N of chlorophyll a, bacteriochlorophyll a, nitrate, and ammonium in benthic microbial mats indicate that inhibition of nitrification suppresses denitrification and anammox, resulting in efficient ammonium recycling within the mats and high primary productivity. We also suggest that the release of 15N-depleted NH3(gas) with increasing salinity enriches ammonium 15N in surface brine (≈34.0‰). Such elevated δ15N is also recorded in geoporphyrins isolated from sediments of the MSC peak (≈20‰), reflecting ammonium supply sufficient for sustaining phototrophic primary production. We propose that efficient nutrient supply combined with frequent bottom-water anoxia and capping of organic-rich sediments by evaporites of the Mediterranean MSC could have contributed to atmospheric CO2 reduction during the late Miocene.

2.
Geobiology ; 8(2): 101-11, 2010 Mar.
Article in English | MEDLINE | ID: mdl-20059556

ABSTRACT

Earth scientists have searched for signs of microscopic life in ancient samples of permafrost, ice, deep-sea sediments, amber, salt and chert. Until now, evidence of cyanobacteria has not been reported in any studies of ancient DNA older than a few thousand years. Here, we investigate morphologically, biochemically and genetically primary evaporites deposited in situ during the late Miocene (Messinian) Salinity Crisis from the north-eastern Apennines of Italy. The evaporites contain fossilized bacterial structures having identical morphological forms as modern microbes. We successfully extracted and amplified genetic material belonging to ancient cyanobacteria from gypsum crystals dating back to 5.910-5.816 Ma, when the Mediterranean became a giant hypersaline brine pool. This finding represents the oldest ancient cyanobacterial DNA to date. Our clone library and its phylogenetic comparison with present cyanobacterial populations point to a marine origin for the depositional basin. This investigation opens the possibility of including fossil cyanobacterial DNA into the palaeo-reconstruction of various environments and could also be used to quantify the ecological importance of cyanobacteria through geological time. These genetic markers serve as biosignatures providing important clues about ancient life and begin a new discussion concerning the debate on the origin of late Miocene evaporites in the Mediterranean.


Subject(s)
Calcium Sulfate , Cyanobacteria/classification , Fossils , Genes, rRNA , Paleontology , RNA, Ribosomal, 16S/genetics , Calcium Sulfate/chemistry , Crystallization , Cyanobacteria/genetics , DNA, Bacterial/analysis , DNA, Bacterial/isolation & purification , Evolution, Molecular , Geologic Sediments/microbiology , Italy , Microscopy, Electron, Scanning , Phylogeny , Sequence Analysis, DNA , Sodium Selenite
3.
Br J Haematol ; 115(3): 554-62, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11736935

ABSTRACT

The localization and establishment of follicular lymphoma (FL) cells in distinct anatomic sites probably involves chemokine and adhesion receptors on the neoplastic cells and appropriate chemokines and adhesion receptor ligands in the microenvironment. Several chemokines play an important role in normal B-cell trafficking and differentiation. Monocyte chemoattractant protein-1 (MCP-1) is a C-C chemokine that induces chemotaxis of a variety of lymphoid cells through its receptor CCR2. CCR2 is also expressed on B cells, and MCP-1 induces chemotaxis of normal B cells. In this report, we investigated expression and function of CCR2 on FL cells. We found FL cells as well as the t(14; 18)+ B-cell lymphoma line H2 expressed CCR2. MCP-1 potentiated SDF-1-induced chemotaxis of FL cells and H2 cells, but MCP-1 alone did not induce chemotaxis. The specificity of the effects of MCP-1 and SDF-1 was demonstrated by antibody blocking studies. Because FL cells are generally associated with follicular dendritic cells (FDCs), FDCs may be an important source of chemokines. We found that cultured FDCs produced MCP-1, and this production was enhanced by tumour necrosis factor. These data implicate MCP-1 in the migration and localization of FL cells.


Subject(s)
Chemokine CCL2/pharmacology , Chemotaxis, Leukocyte/drug effects , Lymphoma, Follicular/metabolism , Receptors, Chemokine/analysis , Antibodies, Monoclonal/pharmacology , Cell Line , Chemokine CCL2/immunology , Chemokine CXCL12 , Chemokines, CXC/immunology , Chemokines, CXC/pharmacology , Chromosomes, Human, Pair 14 , Chromosomes, Human, Pair 18 , Dendritic Cells, Follicular/metabolism , Drug Synergism , Flow Cytometry/methods , Humans , Lymphoma, B-Cell , Lymphoma, Follicular/immunology , Receptors, CCR2 , Receptors, Chemokine/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Translocation, Genetic , Tumor Necrosis Factor-alpha/pharmacology
4.
Cell Immunol ; 203(2): 134-43, 2000 Aug 01.
Article in English | MEDLINE | ID: mdl-11006011

ABSTRACT

Recent studies suggest that tumor necrosis factor (TNF) family members such as TNFalpha and lymphotoxin alphabeta (LTalpha1beta2) are important in the development of follicular dendritic cells (FDCs) and maintenance of FDC function. In this study we used FDC-like cells (FDC-LC) cultured from normal human tonsil and investigated the effects of TNF and LTalpha1beta2 on expression of adhesion molecules and the production of cytokines and chemokines. TNF and LTalpha1beta2 both increased the expression of VCAM-1 and ICAM-1 on FDC-LC. In addition, IL-4 with LTalpha1beta2 synergistically increased the expression of VCAM-1, but not ICAM-1. Cytokine IL-6 and IL-15 mRNAs were induced following stimulation with TNF and LTalpha1beta2. These two cytokines were present in FDC-LC supernatants by ELISA and increased following TNF and LTalpha1beta2 stimulation. We also examined FDC-LC for chemokines, which affect B cells, including IL-8, SDF-1, MIP3beta/ELC, and BCA-1/BLC. SDF-1 mRNA and protein were expressed by FDC-LC, and following stimulation with TNF and LTalpha1beta2, decreases in both were observed. Therefore, TNF and LTalpha1beta2, which are produced by activated B cells, increased the expression of adhesion molecules and cytokines from FDC-LC, potentially providing key signals to support germinal center B cell survival and differentiation.


Subject(s)
Dendritic Cells, Follicular/immunology , Lymphotoxin-alpha/immunology , Tumor Necrosis Factor-alpha/immunology , Cell Line , Chemokines/biosynthesis , Child, Preschool , Cytokines/biosynthesis , Dendritic Cells, Follicular/cytology , Dendritic Cells, Follicular/metabolism , Humans , Intercellular Adhesion Molecule-1/biosynthesis , Tumor Necrosis Factor-alpha/biosynthesis , Vascular Cell Adhesion Molecule-1/biosynthesis
5.
J Cancer Res Clin Oncol ; 125(3-4): 209-13, 1999.
Article in English | MEDLINE | ID: mdl-10235475

ABSTRACT

Three questions associated with the stimulation of cell division by chloride salts have been investigated: (i) whether cations other than sodium show a similar effect, (ii) whether vitamins can have a preventive activity, and (iii) whether subchronic treatment with sodium chloride in the diet is also effective. Male Fischer 344 rats were given solutions of the chloride salts of sodium, potassium, magnesium, and calcium by oral gavage. Water was used for control. After 4 h, a 24-h osmotic minipump containing 5-bromo-2'-deoxyuridine was implanted subcutaneously. The forestomach and glandular stomach, as well as liver and bladder were analyzed immunohistochemically 24 h later for the proportion of cells in S phase as an indicator of the rate of replicative DNA synthesis. For both the forestomach and the glandular stomach, potassium was as potent as sodium, and the divalent cations Mg and Ca were even more potent on a molar basis. Supplementation of the diet with ascorbic acid (2 g/kg food) or beta-carotene (12.5 mg/kg food) for 1 week before gavage of the sodium chloride solution resulted in an inhibition of the stimulation of cell division. A putative tumor-chemopreventive activity of the two vitamins might therefore not only rely on their antioxidative properties but may include effects on the cell cycle. A 4-week treatment with a sodium chloride supplement in the diet (2% and 4% supplement) resulted in a significant stimulation of cell division not only in both parts of the stomach and in the bladder (with the 4% supplement) but also in the liver (even with the 2% supplement). Sodium-chloride-stimulated cell turnover therefore is a sustained effect.


Subject(s)
Antioxidants/pharmacology , Ascorbic Acid/pharmacology , Chlorides/pharmacology , Sodium Chloride/antagonists & inhibitors , Stomach/cytology , Stomach/drug effects , beta Carotene/pharmacology , Animals , Bromodeoxyuridine/metabolism , Calcium Chloride/pharmacology , Cell Division/drug effects , DNA/metabolism , Immunohistochemistry , Magnesium Chloride/pharmacology , Male , Potassium Chloride/pharmacology , Rats , Rats, Inbred F344 , S Phase/drug effects , S Phase/physiology , Sodium Chloride/pharmacology
6.
Lab Invest ; 78(5): 591-602, 1998 May.
Article in English | MEDLINE | ID: mdl-9605184

ABSTRACT

Interleukin (IL)-4 and IL-13 are known to bind to shared heteromultimeric receptor complexes of variable composition. Given the many regulatory effects of IL-4 and IL-13 on synovial cells, we aimed to characterize their IL-4/IL-13 receptor (R). Cultivated synovial fibroblasts expressed transcripts for IL-4Ralpha and IL-13Ralpha1, the human homolog of the recently cloned mouse IL-13R, but not the common gamma-chain of the IL-2R. In particular, IL-13Ralpha2 mRNA, encoding a different IL-13R recently cloned from human renal carcinoma cells, was expressed at a strikingly high level. Correspondingly, a predominant protein migrating at 65 to 75 kd was cross-linked by iodinated IL-13 and was not cross-competed by an excess of unlabeled IL-4. However, by flow cytofluorometry, IL-13Ralpha1 (detected by the anti-lL-13Ralpha1 mAb 65) and IL-4Ralpha (detected by the mAb S697) were expressed at similar low density. Radioligand binding studies revealed for both cytokines approximately 300 receptors/cell with similar high affinity. An additional class of IL-13Rs was identified after occupation of the shared high-affinity receptors by the nonsignaling, double-mutant IL-4121R-->D, 124Y-->D (RY-IL-4). In these experiments, 1251-IL-13 bound to a single receptor population with a Kd of approximately 300 pM and approximately 5000 sites/cell, matching the published affinity of monomeric IL-13Ralpha2 when expressed in COS7 cells. RY-IL-4 blocked the IL-4- and IL-13-mediated vascular cell adhesion molecule (VCAM)-1 expression and Stat6 activation, suggesting that the large number of high-affinity IL-13Ralpha2 monomers are silent receptors, likely representing a decoy target for IL-13.


Subject(s)
Receptors, Interleukin-4/metabolism , Receptors, Interleukin/metabolism , Synovial Membrane/metabolism , Carrier Proteins/metabolism , Fibroblasts/metabolism , Humans , Interleukin-13/metabolism , Interleukin-13/pharmacology , Interleukin-13 Receptor alpha1 Subunit , Interleukin-4/metabolism , Interleukin-4/pharmacology , RNA, Messenger/metabolism , Receptors, Interleukin/genetics , Receptors, Interleukin-13 , Receptors, Interleukin-4/genetics , Signal Transduction/physiology , Synovial Membrane/cytology , Vascular Cell Adhesion Molecule-1/metabolism
7.
Fundam Appl Toxicol ; 39(2): 131-7, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9344625

ABSTRACT

Caffeic acid (CA, 3,4-dihydroxycinnamic acid), at 2% in the diet, had been shown to be carcinogenic in forestomach and kidney of F344 rats and B6C3F1 mice. Based on its occurrence in coffee and numerous foods and using a linear interpolation for cancer incidence between dose 0 and 2%, the cancer risk in humans would be considerable. In both target organs, tumor formation was preceded by hyperplasia, which could represent the main mechanism of carcinogenic action. The dose-response relationship for this effect was investigated in male F344 rats after 4-week feeding with CA at different dietary concentrations (0, 0.05, 0.14, 0.40, and 1.64%). Cells in S-phase of DNA replication were visualized by immunohistochemical analysis of incorporated 5-bromo-2'-deoxyuridine (BrdU), 2 hr after intraperitoneal injection. In the forestomach, both the total number of epithelial cells per millimeter section length and the unit length labeling index of BrdU-positive cells (ULLI) were increased, about 2.5-fold, at 0.40 and 1.64%. The lowest concentration (0.05%) had no effect. At 0.14%, both variables were decreased by about one-third. In the kidney, the labeling index in proximal tubular cells also indicated a J-shaped (or U-shaped) dose response with a 1. 8-fold increase at 1.64%. In the glandular stomach and in the liver, which are not target organs, no dose-related effect was seen. The data show a good correlation between the organ specificity for cancer induction and stimulation of cell division. With respect to the dose-response relationship and the corresponding extrapolation of the animal tumor data to a human cancer risk, a linear extrapolation appears not to be appropriate.


Subject(s)
Antioxidants/toxicity , Caffeic Acids/toxicity , Carcinogens/toxicity , Kidney/drug effects , Stomach/drug effects , Animal Feed , Animals , Body Weight/drug effects , Bromodeoxyuridine/metabolism , Caffeic Acids/administration & dosage , Cell Division/drug effects , Dose-Response Relationship, Drug , Eating , Hyperplasia/chemically induced , Immunohistochemistry , Kidney/pathology , Kidney Tubules, Proximal/drug effects , Kidney Tubules, Proximal/pathology , Male , Pilot Projects , Rats , Rats, Inbred F344 , S Phase , Stomach/pathology
8.
J Biol Chem ; 272(9): 5487-94, 1997 Feb 28.
Article in English | MEDLINE | ID: mdl-9038152

ABSTRACT

Functional receptors for interleukin (IL)-4 and IL-13 on endothelial cells consist of the 130-kDa IL-4 receptor alpha-chain (IL-4Ralpha) and a 65-75-kDa IL-13 binding subunit that are expressed in a ratio of about 1:3, respectively. The restricted number of IL-4Ralpha limits subunit heterodimerization and in turn receptor-mediated signaling. We report here, the effects of tumor necrosis factor alpha (TNF-alpha) on the expression of the receptor subunits for IL-4 and IL-13. By flow cytofluorometry and receptor-binding analysis of iodinated IL-4 and IL-13, stimulation with TNF-alpha-induced a 2-3-fold increase of the IL-4Ralpha expression. The up-regulation was also confirmed at the transcriptional level by reverse transcription-polymerase chain reaction. Radioligand cross-linking experiments revealed no change in the subunit composition of the TNF-alpha-induced receptor complex. Nevertheless, TNF-alpha stimulation led to increased activation of the IL-4-specific signal transducers and activators of transcription protein (Stat6) by IL-4 and IL-13. Thus, TNF-alpha corrects the subunit imbalance of the endothelial IL-4.IL-13 receptor complex thereby increasing receptor heterodimerization and in turn the signaling capability by IL-4 and IL-13.


Subject(s)
Antigens, CD/metabolism , Interleukin-13/metabolism , Interleukin-4/metabolism , Receptors, Interleukin/metabolism , Signal Transduction , Trans-Activators/metabolism , Tumor Necrosis Factor-alpha/pharmacology , Animals , Cricetinae , Cricetulus , Electrophoresis, Polyacrylamide Gel , Endothelium, Vascular/metabolism , Enzyme Activation , Female , Humans , Models, Molecular , Polymerase Chain Reaction , Protein-Tyrosine Kinases/metabolism , Receptors, Interleukin-4 , STAT6 Transcription Factor , Spectrometry, Fluorescence , Up-Regulation
9.
Am J Pathol ; 149(4): 1369-79, 1996 Oct.
Article in English | MEDLINE | ID: mdl-8863684

ABSTRACT

The distribution of the interleukin (IL)-4 receptor in normal human and common marmoset (Callithrix jacchus) tissues was examined by immunofluorescence and flow cytometry using monoclonal antibodies specific for the human IL-4 receptor to gain further insight into IL-4-mediated inflammatory and immunological events. IL-4 receptor positivity was unequivocally demonstrated on lymphocytes, predominantly T cells, and on blood vessels in many tissues. Vascular IL-4 receptor immunofluorescence consisted of a strong smooth muscle cell positivity and weaker positive staining of capillary and venular endothelial cells. Subnanomolar concentrations of IL-4 induced a genistein-sensitive up-regulation of VCAM-1 in vascular cell cultures. Tumor necrosis factor-alpha induced a genistein-resistant up-regulation of VCAM-1. IL-4 strongly induced expression of the IL-4 receptor on splenocytes (T lymphocytes) but not on vascular smooth muscle or endothelial cell cultures. Receptor cross-linking to [125I]IL-4 revealed a 65- to 75-kDa accessory receptor subunit consistent with a recently cloned IL-13 receptor associated with the IL-4 receptor on both vascular endothelial and smooth muscle cells. The demonstration of a vascular distribution pattern for the IL-4 receptor in addition to expression on lymphocytes suggests that vascular functional alterations, transduced through a unique IL-4 receptor complex (the type II IL-4 receptor), may be of importance during immunological and allergic inflammatory events.


Subject(s)
Antigens, CD/analysis , Antigens, CD/physiology , Endothelium, Vascular/chemistry , Lymphocytes/chemistry , Muscle, Smooth/chemistry , Receptors, Interleukin/analysis , Receptors, Interleukin/physiology , Animals , Antibodies, Monoclonal , Antigens, CD/chemistry , Callithrix , Endothelium, Vascular/metabolism , Enzyme Inhibitors/pharmacology , Genistein , Humans , Isoflavones/pharmacology , Muscle, Smooth/metabolism , Receptors, Interleukin/chemistry , Receptors, Interleukin-4 , Vascular Cell Adhesion Molecule-1/drug effects , Vascular Cell Adhesion Molecule-1/metabolism
10.
Blood ; 87(10): 4286-95, 1996 May 15.
Article in English | MEDLINE | ID: mdl-8639787

ABSTRACT

Interleukin-4 (IL-4) and IL-13 exert similar, nonadditive effects on endothelial cells, inducing vascular cell adhesion molecule-1 (VCAM-1) expression and subsequent transmigration of eosinophils. The receptor for IL-4 and IL-13 was described as a shared heteromultimeric complex in which the common gamma-chain (gamma c) subunit was essential for activity. Endothelial cell bound both cytokines with high affinity; by flow cytofluorometry and reverse transcription-polymerase chain reaction (RT-PCR), they expressed IL-4 receptor alpha (IL-4R alpha) but did not express the gamma c of the IL-2R. Radioligand cross-linking experiments followed by immunoprecipitation with the monoclonal antibody (MoAb) S697 to the IL-4R alpha showed IL-4-specific binding at 130 kD, the IL-4R alpha, and to a minor extent to a double band coimmunoprecipitated at 65 to 75 kD. [125 I]IL-13 bound predominantly to the 65- to 75- kD band and with a trace amount of binding at 130 kD. However, no ligand-cross-linked receptor was precipitated by the MoAb S697, indicating a cognate novel IL-13-binding subunit. Excess unlabeled IL-4 completely displaced IL-13 binding. Similarly, nonsignaling IL-4 (Y124D)-mutant abolished IL-4- and IL-13-mediated signal transduction. Unlabeled IL-13 competed successfully for IL-4 binding at 65 to 75 kD but was unable to completely displace Il-4 from its binding to the IL-4R alpha. The MoAb TUGh4, specific for the gamma c, failed to precipitate ligand-cross-linked IL-4R and IL-13R. Therefore, the subunit structure of the functional receptors for IL-4 and IL-13 on human endothelial cells does not use or require the common gamma c of the IL-2R.


Subject(s)
Antigens, CD/metabolism , Endothelium, Vascular/metabolism , Gene Expression Regulation/drug effects , Interleukin-13/metabolism , Interleukin-4/metabolism , Receptors, Interleukin/metabolism , Vascular Cell Adhesion Molecule-1/biosynthesis , Amino Acid Sequence , Animals , Antigens, CD/chemistry , Antigens, CD/genetics , Base Sequence , Binding, Competitive , CHO Cells , Cell Line, Transformed , Cricetinae , Cricetulus , Endothelium, Vascular/drug effects , Enzyme Inhibitors/pharmacology , Humans , Interleukin-13/pharmacology , Interleukin-13 Receptor alpha1 Subunit , Interleukin-4/analogs & derivatives , Interleukin-4/genetics , Interleukin-4/pharmacology , Molecular Sequence Data , Polymerase Chain Reaction , Protein Kinase Inhibitors , Receptors, Interleukin/chemistry , Receptors, Interleukin/genetics , Receptors, Interleukin-13 , Receptors, Interleukin-2/biosynthesis , Receptors, Interleukin-2/chemistry , Receptors, Interleukin-2/genetics , Receptors, Interleukin-4 , Signal Transduction , Vascular Cell Adhesion Molecule-1/genetics
11.
J Exp Med ; 181(5): 1893-8, 1995 May 01.
Article in English | MEDLINE | ID: mdl-7722464

ABSTRACT

Interleukin (IL)-12 synergizes with other cytokines to stimulate the proliferation and differentiation of early hematopoietic progenitors in vitro. However, in vivo administration of IL-12 decreases peripheral blood counts and bone marrow hematopoiesis. Here, we used interferon (IFN) gamma receptor-deficient (IFN gamma R-/-) mice to investigate whether the in vivo inhibition of hematopoiesis by IL-12 is indirectly mediated by IL-12-induced IFN-gamma. IL-12 administered for 4 d (1 microgram/mouse per day) resulted in lower peripheral blood counts and a 2-fold decrease in bone marrow cellularity in wild-type mice, but not in IFN gamma R-/- mice. Bone marrow hematopoietic progenitors were decreased after IL-12 treatment in wild-type mice, but rather increased in IFN gamma R-/- mice. Splenic cellularity was 2.3-fold higher after IL-12 administration in wild-type mice, largely due to natural killer (NK) cell and macrophage infiltration together with some extramedullary hematopoiesis. In IFN gamma R-/- mice, spleen cellularity was less increased, there were fewer infiltrating NK cells, but a strong extramedullary hematopoiesis. Thus, alterations mediated by IL-12-induced IFN-gamma include reduction in bone marrow cellularity and hematopoietic progenitors, as well as pronounced splenomegaly, largely caused by NK cell infiltration. In the absence of IFN-gamma signaling, IL-12 promotes hematopoiesis, consistent with its in vitro activities.


Subject(s)
Hematopoiesis/drug effects , Interferon-gamma/physiology , Interleukin-12/antagonists & inhibitors , Animals , Bone Marrow/drug effects , Mice , Receptors, Interferon/analysis , Interferon gamma Receptor
13.
Minerva Chir ; 34(20): 1369-76, 1979 Oct 31.
Article in Italian | MEDLINE | ID: mdl-159421

ABSTRACT

A diagnostic assessment was made of 74 cases of non-penetrating abdominal wounds observed over the previous 5 years. Laparoscopy performed in 49 polytraumatised patients made a decisive contribution to accurate, early diagnosis, with an indication for emergency surgery in 35 cases (32 visceral lesions and 3 retroperitoneal haematoma), whereas it was clear from the negative finding that surgery was not required in the remaining 14. It is felt that all additional radiological and instrumental diagnostic should be employed, and that careful clinical examination and repeated inspection, preferably by the same surgeon, are important in cases of multiple injury, since the situation may become dramatically worse in the space of a few hours.


Subject(s)
Abdominal Injuries/diagnosis , Abdominal Injuries/diagnostic imaging , Abdominal Injuries/surgery , Adolescent , Adult , Aged , Child , Female , Humans , Laparoscopy , Male , Middle Aged , Radiography
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