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The Triterpenoid Cddo Inhibits Expression Of Matrix Metalloproteinase-1, Matrix Metalloproteinase-13 And Bcl-3 In Primary Human Chondrocytes, Sarah Elliott, Ezra Hays, Michael Mayor, Michael Sporn, Matthew P. Vincenti

Dartmouth Scholarship

A synthetic triterpenoid, 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO), has been reported to have anti-inflammatory properties and to decrease the interleukin-1 (IL-1)-induced expression of matrix metalloproteinase-1 (MMP-1) and MMP-13. We have shown previously that IL-1 induces expression of the inhibitor of NF-κB (IκB) family member Bcl-3, and that this contributes to MMP-1 expression. To quantify the effects of CDDO on IL-1-induced MMP-1, MMP-13 and Bcl-3 expression, we stimulated the chondrosarcoma cell line SW-1353 and human primary chondrocytes with IL-1, in the presence or absence of CDDO. Harvested RNA was subjected to quantitative real-time reverse-transcriptase polymerase chain reaction. In SW-1353 cells, 300 nM CDDO …

Compromised Constitutions: The Iranian Experience With The 1918 Influenza Pandemic., Amir Afkhami

Psychiatry and Behavioral Sciences Faculty Publications

The global demographic impact of the 1918-19 influenza pandemic continues to fascinate researchers and scholars. This paper examines the social and demographic effects of this outbreak on Iranian society, through a comprehensive investigation of the modes of transmission and propagation, mortality rates, and other distinctive features of the region, and reveals the importance of taking a country's unique sociopolitical settings into account. Iran was one of the regions hit hardest by the pandemic, with mortality rates significantly higher than in most regions of the world. Though globally the victims of influenza lived primarily in urban areas, it was Iran's rural …

Crystal Structure Of The Sars Protein From Staphylococcus Aureus, Ronggui Li, Adhar C. Manna, Shaodong Dai, Ambrose L. Cheung, Gongyi Zhang

Dartmouth Scholarship

The expression of virulence determinants in Staphylococcus aureus is controlled by global regulatory loci (e.g., sarA and agr). One of these determinants, protein A (spa), is activated by sarS, which encodes a 250-residue DNA-binding protein. Genetic analysis indicated that the agr locus likely mediates spa repression by suppressing the transcription of sarS. Contrary to SarA and SarR, which require homodimer formation for proper function, SarS is unusual within the SarA protein family in that it contains two homologous halves, with each half sharing sequence similarity to SarA and SarR. Here we report the 2.2 Å …

Children With Disturbances In Sensory Processing: A Pilot Study Examining The Role Of The Parasympathetic Nervous System., Roseann C. Schaaf, Lucy Jane Miller, Duncan Seawell, Shannon O'Keefe

Department of Occupational Therapy Faculty Papers

This study was a preliminary investigation of parasympathetic nervous system (PNS) functioning in children with disturbances in sensory processing. The specific aims of this study were to (1) provide preliminary data about group differences in parasympathetic functions, as measured by the vagal tone index, between children with disturbances in sensory processing and those without; (2) determine effect size and power needed for future studies; and (3) to lay the foundation for further examination of the relations of parasympathetic functioning and functional behavior in children with disturbances in sensory processing. Participants were 15 children, nine with disturbances in sensory processing and …

A Functional Relationship Between Numa And Kid Is Involved In Both Spindle Organization And Chromosome Alignment In Vertebrate Cells, Aime A. Levesque, Louisa Howard, Michael B. Gordon, Duane A. Compton

Dartmouth Scholarship

We examined spindle morphology and chromosome alignment in vertebrate cells after simultaneous perturbation of the chromokinesin Kid and either NuMA, CENP-E, or HSET. Spindle morphology and chromosome alignment after simultaneous perturbation of Kid and either HSET or CENP-E were no different from when either HSET or CENP-E was perturbed alone. However, short bipolar spindles with organized poles formed after perturbation of both Kid and NuMA in stark contrast to splayed spindle poles observed after perturbation of NuMA alone. Spindles were disorganized if Kid, NuMA, and HSET were perturbed, indicating that HSET is sufficient for spindle organization in the absence of …

Probucol Prevents Early Coronary Heart Disease And Death In The High-Density Lipoprotein Receptor Sr-Bi/Apolipoprotein E Double Knockout Mouse, Anne Braun, Songwen Zhang, Helena E. Miettinen, Shamsah Ebrahim, Teresa M. Holm, Eliza Vasile, Mark J. Post

Dartmouth Scholarship

Mice with homozygous null mutations in the high-density lipoprotein receptor SR-BI (scavenger receptor class B, type I) and apolipoprotein E genes fed a low-fat diet exhibit a constellation of pathologies shared with human atherosclerotic coronary heart disease (CHD): hypercholesterolemia, occlusive coronary atherosclerosis, myocardial infarctions, cardiac dysfunction (heart enlargement, reduced systolic function and ejection fraction, and ECG abnormalities), and premature death (mean age 6 weeks). They also exhibit a block in RBC maturation and abnormally high plasma unesterified-to-total cholesterol ratio (0.8) with associated abnormal lipoprotein morphology (lamellar/vesicular and stacked discoidal particles reminiscent of those in lecithin/cholesterol acyltransferase deficiency and cholestasis). Treatment …

Copper Chelation Represses The Vascular Response To Injury, Lazar Mandinov, Anna Mandinova, Stanimir Kyurkchiev, Dobroslav Kyurkchiev, Ivan Kehayov, Vihren Kolev, Raffaella Soldi, Cinzia Bagala, Ebo D. De Muinck, Volkhard Lindner, Mark J. Post, Michael Simons

Dartmouth Scholarship

The induction of an acute inflammatory response followed by the release of polypeptide cytokines and growth factors from peripheral blood monocytes has been implicated in mediating the response to vascular injury. Because the Cu2+-binding proteins IL-1alpha and fibroblast growth factor 1 are exported into the extracellular compartment in a stress-dependent manner by using intracellular Cu2+ to facilitate the formation of S100A13 heterotetrameric complexes and these signal peptideless polypeptides have been implicated as regulators of vascular injury in vivo, we examined the ability of Cu2+ chelation to repress neointimal thickening in response to injury. We observed that the oral administration of …

Is Hp1 An Rna Detector That Functions Both In Repression And Activation?, Rebecca Kellum

Biology Faculty Publications

Heterochromatin is defined as regions of compact chromatin that persist throughout the cell cycle (Heitz, 1928). The earliest cytological observations of heterochromatin were followed by ribonucleotide labeling experiments that showed it to be transcriptionally inert relative to the more typical euchromatic regions that decondense during interphase. Genetic studies of rearrangements that place euchromatic genes next to blocks of heterochromatin also pointed out the repressive nature of heterochromatin (Grigliatti, 1991; and references therein). The discovery of the heterochromatin-enriched protein heterochromatin protein 1 (HP1) by Elgin and co-workers in the mid-1980s suggested that the distinct cytological features of this chromatin may be …

Regulation And Localization Of Endogenous Human Tristetraprolin, Anna-Marie Fairhurst, John E. Connolly, Katharine A Hintz, Nicolas J Goulding

Dartmouth Scholarship

Tumor necrosis factor (TNF) has been implicated in the development and pathogenicity of infectious diseases and autoimmune disorders, such as septic shock and arthritis. The zinc-finger protein tristetraprolin (TTP) has been identified as a major regulator of TNF biosynthesis. To define its intracellular location and examine its regulation of TNF, a quantitive intracellular staining assay specific for TTP was developed. We establish for the first time that in peripheral blood leukocytes, express

Genomic And Proteomic Profiling Of Responses To Toxic Metals In Human Lung Cells, Angeline S. Andrew, Amy J. Warren, Aaron Barchowsky, Kaili A. Temple, Linda Klei, Nicole V. Soucy, Kimberly A. O'Hara, Joshua W. Hamilton

Dartmouth Scholarship

Examining global effects of toxic metals on gene expression can be useful for elucidating patterns of biological response, discovering underlying mechanisms of toxicity, and identifying candidate metal-specific genetic markers of exposure and response. Using a 1,200 gene nylon array, we examined changes in gene expression following low-dose, acute exposures of cadmium, chromium, arsenic, nickel, or mitomycin C (MMC) in BEAS-2B human bronchial epithelial cells. Total RNA was isolated from cells exposed to 3 M Cd(II) (as cadmium chloride), 10 M Cr(VI) (as sodium dichromate), 3 g/cm2 Ni(II) (as nickel subsulfide), 5 M or 50 M As(III) (as sodium arsenite), or …

Mammalian Erv46 Localizes To The Endoplasmic Reticulum–Golgi Intermediate Compartment And To Cis-Golgi Cisternae, Lelio Orci, Mariella Ravazzola, Gary J. Mack, Charles Barlowe, Stefan Otte

Dartmouth Scholarship

Yeast endoplasmic reticulum (ER) vesicle protein Erv46p is a novel membrane protein involved in transport through the early secretory pathway. Investigation of mammalian Erv46 (mErv46) reveals that it is broadly expressed in tissues and protein-secreting cells. By immunofluorescence microscopy, mErv46 displays a crescent-shaped perinuclear staining pattern that is characteristic of the Golgi complex. Quantitative immunoelectron microscopy indicates that mErv46 is restricted to the cis face of the Golgi apparatus and to vesicular tubular structures between the transitional ER and cis-Golgi. Minor amounts of mErv46 reside in ER membranes and later Golgi cisternae. On Brefeldin A treatment, mErv46 redistributes to punctate …

Cell-Cycle Regulatory Proteins In Podocyte Cell In Idiopathic Nephrotic Syndrome Of Childhood., Tarak Srivastava, Robert E. Garola, J M. Whiting, Uri S. Alon

Manuscripts, Articles, Book Chapters and Other Papers

BACKGROUND: The podocyte cell is believed to play an important role in idiopathic nephrotic syndrome (INS) of childhood. In adults with cellular and collapsing focal segmental glomerulosclerosis (FSGS), the expression of cell-cycle regulatory proteins such as p27, p57, and cyclin D is decreased and expression of cyclin A, Ki-67, and p21 is observed in podocyte cells suggestive of a dysregulated podocyte phenotype. We investigated for alterations in the expression of cyclin kinase inhibitors, p27, p57, p21, and cyclins D and A in the podocyte cell of children with INS.

METHODS: Forty-two kidney biopsies were investigated; 14 with minimal-change disease (MCD), …

Human Acyl-Coenzyme A:Cholesterol Acyltransferase Expressed In Chinese Hamster Ovary Cells: Membrane Topology And Active Site Location, Song Lin, Xiaohui Lu, Catherine C.Y. Chang, Ta-Yuan Chang

Dartmouth Scholarship

Acyl-CoA:cholesterol acyltransferase (ACAT) is a membrane-bound enzyme that produces cholesteryl esters intracellularly. Two ACAT genes (ACAT1 and ACAT2) have been identified. The expression of ACAT1 is ubiquitous, whereas that of ACAT2 is tissue restricted. Previous research indicates that ACAT1 may contain seven transmembrane domains (TMDs). To study ACAT2 topology, we inserted two different antigenic tags (hemagglutinin, monoclonal antibody Mab1) at various hydrophilic regions flanking each of its predicted TMDs, and expressed the recombinant proteins in mutant Chinese hamster ovary cells lacking endogenous ACAT. Each tagged ACAT2 was expressed in the endoplasmic reticulum as a single undegraded protein band …

Minus-End Capture Of Preformed Kinetochore Fibers Contributes To Spindle Morphogenesis, Alexey Khodjakov, Lily Copenagle, Michael B. Gordon, Duane A. Compton, Tarun M. Kapoor

Dartmouth Scholarship

Near-simultaneous three-dimensional fluorescence/differential interference contrast microscopy was used to follow the behavior of microtubules and chromosomes in living alpha-tubulin/GFP-expressing cells after inhibition of the mitotic kinesin Eg5 with monastrol. Kinetochore fibers (K-fibers) were frequently observed forming in association with chromosomes both during monastrol treatment and after monastrol removal. Surprisingly, these K-fibers were oriented away from, and not directly connected to, centrosomes and incorporated into the spindle by the sliding of their distal ends toward centrosomes via a NuMA-dependent mechanism. Similar preformed K-fibers were also observed during spindle formation in untreated cells. In addition, upon monastrol removal, centrosomes established a transient …

Glucuronidation Enzymes, Genes And Psychiatry, Jose De Leon

Psychiatry Faculty Publications

The phase I cytochrome P450 (CYP) isoenzymes have received substantial attention in the pharmacogenetic literature. Researchers are beginning to examine the role of the phase II UDP-glucuronosyltransferase (UGT) enzymes, which produce products that are more water-soluble, less toxic and more readily excreted than the parent compounds. Several reasons may have contributed to neglect of UGTs (compared to CYPs) including: (1) the overlapping activity of UGTs and lack of selective probes; (2) the complexity of the glucuronidation cycle; and (3) the difficulty in developing analytic methods to measure glucuronides. Current CYP knowledge is used as a model to predict advances in …

Transfection Of Il-10 Expression Vectors Into Endothelial Cultures Attenuates Alpha4beta7-Dependent Lymphocyte Adhesion Mediated By Madcam-1., Makoto Sasaki, Paul Jordan, Jeff Houghton, Xianmin Meng, Makoto Itoh, Takashi Joh, J Steven Alexander

Department of Dermatology and Cutaneous Biology Faculty Papers

BACKGROUND: Enhanced expression of MAdCAM-1 (mucosal addressin cell adhesion molecule-1) is associated with the onset and progression of inflammatory bowel disease. The clinical significance of elevated MAdCAM-1 expression is supported by studies showing that immunoneutralization of MAdCAM-1, or its ligands reduce inflammation and mucosal damage in models of colitis. Interleukin-10 (IL-10) is an endogenous anti-inflammatory and immunomodulatory cytokine that has been shown to prevent inflammation and injury in several animal studies, however clinical IL-10 treatment remains insufficient because of difficulties in the route of IL-10 administration and its biological half-life. Here, we examined the ability of introducing an IL-10 expression …

Responsiveness Of Bovine Cumulus-Oocyte-Complexes (Coc) To Porcine And Recombinant Human Fsh, And The Effect Of Coc Quality On Gonadotropin Receptor And Cx43 Marker Gene Mrnas During Maturation In Vitro., Michele D Calder, Anita N Caveney, Lawrence C Smith, Andrew J Watson

Obstetrics & Gynaecology Publications

Substantially less development to the blastocyst stage occurs in vitro than in vivo and this may be due to deficiencies in oocyte competence. Although a large proportion of bovine oocytes undergo spontaneous nuclear maturation, less is known about requirements for proper cytoplasmic maturation. Commonly, supraphysiological concentrations of FSH and LH are added to maturation media to improve cumulus expansion, fertilization and embryonic development. Therefore, various concentrations of porcine FSH (pFSH) and recombinant human FSH (rhFSH) were investigated for their effect on bovine cumulus expansion in vitro. Expression of FSHr, LHr and Cx43 mRNAs was determined in cumulus-oocyte complexes to determine …

Promoters Of The Murine Embryonic Β-Like Globin Genes Ey And Βh1 Do Not Compete For Interaction With The Β-Globin Locus Control Region, Xiao Hu, Michael Bulger, Julia N. Roach, Susan K. Eszterhas, Emmanuel Olivier, Eric Bouhassira, Mark Groudine, Steven Fiering

Dartmouth Scholarship

Mammalian β-globin loci contain multiple β-like genes that are expressed at different times during development. The murine β-globin locus contains two genes expressed during the embryo stage, Ey and βh1, and two genes expressed at both the fetal and postnatal stages, β-major and β-minor. Studies of transgenic human β-like globin loci in mice have suggested that expression of one gene at the locus will suppress expression of other genes at the locus. To test this hypothesis we produced mouse lines with deletions of either the Ey or βh1 promoter in the endogenous murine β-globin locus. Promoter deletion eliminated expression of …

Effects Of Tacrolimus On Ischemia-Reperfusion Injury., Shawn D. St Peter, Adyr A. Moss, David C. Mulligan

Manuscripts, Articles, Book Chapters and Other Papers

In addition to efficacious immunosuppression for the benefit of organ transplantation, tacrolimus has diverse actions that result in amelioration of ischemia-reperfusion injury. Knowledge is accumulating rapidly on the mechanisms through which tacrolimus exerts these cytoprotective effects, including alterations in microcirculation, free radical metabolism, calcium-activated pathways, inflammatory cascades, mitochondrial stability, apoptosis, stress-response proteins, and tissue recovery. Within the nucleus, actions mediating the effects of tacrolimus appear to be dominantly influenced by interactions with the transcription factor, nuclear factor-kappaB. Because tacrolimus is a cornerstone agent in immunosuppression regimens throughout the world and knowledge of its cellular mechanisms is evolving, it is important …

Might A Psychosocial Approach Improve Our Understanding Of Itching And Scratching?, Herbert M. Adler

Department of Psychiatry and Human Behavior Faculty Papers

No abstract provided.

Regulation Of Human Col9a1 Gene Expression. Activation Of The Proximal Promoter Region By Sox9., Ping Zhang, Sergio A. Jimenez, David G Stokes

Department of Medicine Faculty Papers

The COL9A1 gene contains two promoter regions, one driving expression of a long alpha1(IX) chain in cartilage (upstream) and one driving expression of a shorter chain in the cornea and vitreous (downstream). To determine how the chondrocyte-specific expression of the COL9A1 gene is regulated, we have begun to characterize the upstream chondrocyte-specific promoter region of the human COL9A1 gene. Transient-transfection analyses performed in rat chondrosarcoma (RCS) cells, human chondrosarcoma (HTB) cells, and NIH/3T3 cells showed that the COL9A1 promoter was active in RCS cells but not HTB or NIH/3T3 cells. Inclusion of the first intron had no effect on promoter …

The Permeability Of The Endoplasmic Reticulum Is Dynamically Coupled To Protein Synthesis, Anirban Roy, William F. Wonderlin

Faculty & Staff Scholarship

No abstract provided.

Nucleotide Excision Repair- And Polymerase Eta-Mediated Error-Prone Removal Of Mitomycin C Interstrand Cross-Links, H. Zheng, X. Wang, A. J. Warren, R. J. Legerski, Rodney S. Nairn, Joshua W. Hamilton, Lei Li

Dartmouth Scholarship

Interstrand cross-links (ICLs) make up a unique class of DNA lesions in which both strands of the double helix are covalently joined, precluding strand opening during replication and transcription. The repair of DNA ICLs has become a focus of study since ICLs are recognized as the main cytotoxic lesion inflicted by an array of alkylating compounds used in cancer treatment. As is the case for double-strand breaks, a damage-free homologous copy is essential for the removal of ICLs in an error-free manner. However, recombination-independent mechanisms may exist to remove ICLs in an error-prone fashion. We have developed an in vivo …

Consumption Of Omega-3 Fatty Acid-Enriched Eggs And Serum Lipids In Humans, Ji-Young Lee, Nancy M. Lewis, Sheila Scheideler, Timothy P. Carr

Department of Nutrition and Health Sciences: Faculty Publications

This study examined the effectiveness of consuming omega-3 fatty acid-enriched eggs (Omega Eggs) in increasing total diclary omega-3 fatty acids. Also examined was the impact of Omega Egg consumption on serum lipids. Sixteen hypercholesterolemic men and women with baseline serum total cholesterol concentrations of 5.17-7.76 mmol/L (200-300 mg/dL) followed the National Cholesterol Education Program Step I diet guidelines under the following conditions: (a) Step 1 diet without eggs, (b) Step I diet plus 12 regular eggs per week, and (c) Step I diet plus 12 Omega Eggs per week. The study design was a repeated 3 x 3 Latin square …

Family Connectedness And Sexual Risk-Taking Among Urban Youth Attending Alternative High Schools, Christine M Markham, Susan R Tortolero, S Liliana Escobar-Chaves, Guy S Parcel, Ronald Harrist, Robert C Addy

Faculty, Staff and Student Publications

CONTEXT: Youth in alternative high schools engage in risky sexual behavior at higher rates than do their peers in regular schools, placing themselves at an increased risk of sexually transmitted disease and unintended pregnancy. Family connectedness is associated with reduced adolescent sexual risk-taking, although this association has not been tested among alternative school youth.

METHODS: A sample of 976 urban, predominantly minority alternative high school students in Houston, Texas, were surveyed in 2000-2002. Survey data were analyzed using logistic regression to determine whether family connectedness is related to sexual risk-taking.

RESULTS: Overall, 68% of students reported ever having had sex. …

Plasma Pge-2 Levels And Altered Cytokine Profiles In Adherent Peripheral Blood Mononuclear Cells In Non-Small Cell Lung Cancer (Nsclc), Giovanna E. Hidalgo, Li Zhong, Dennis E. Doherty, Edward A. Hirschowitz

Internal Medicine Faculty Publications

INTRODUCTION: PGE-2 is constitutively produced by many non-small cell lung cancers (NSCLC) and its immunosuppressive effects have been linked to altered immune responses in lung cancer. We asked whether elevated levels of plasma PGE-2 correlated with monocyte IL10 production in the NSCLC environment. Looking for correlation in NSCLC patient blood we assayed plasma from NSCLC patients for PGE2 and IL10; we further evaluated production of IL10 by adherent mononuclear cells from a subset of these patients looking for an altered cytokine profile.

RESULTS: Our initial in vitro experiments show that monocyte IL10 induction correlates with tumor cell PGE-2 production, confirming …

Distinct Neural Systems Subserve Person And Object Knowledge, Jason P. Mitchell, Todd F. Heatherton, C. Neil Macrae

Dartmouth Scholarship

Studies using functional neuroimaging and patient populations have demonstrated that distinct brain regions subserve semantic knowledge for different classes of inanimate objects (e.g., tools, musical instruments, and houses). What this work has yet to consider, however, is how conceptual knowledge about people may be organized in the brain. In particular, is there a distinct functional neuroanatomy associated with person knowledge? By using event-related functional magnetic resonance imaging (fMRI), we measured neural activity while participants made semantic judgments about people or objects. A unique pattern of brain activity was associated with person judgments and included brain regions previously implicated in other …

Mad2 And Bubr1 Function In A Single Checkpoint Pathway That Responds To A Loss Of Tension, Katie Shannon, Julie C. Canman, Edward D. Salmon

Biological Sciences Faculty Research & Creative Works

The spindle checkpoint monitors microtubule attachment and tension at kinetochores to ensure proper chromosome segregation. Previously, PtK1 cells in hypothermic conditions (23°C) were shown to have a pronounced mitotic delay, despite having normal numbers of kinetochore microtubules. At 23°C, we found that PtK1 cells remained in metaphase for an average of 101 min, compared with 21 min for cells at 37°C. The metaphase delay at 23°C was abrogated by injection of Mad2 inhibitors, showing that Mad2 and the spindle checkpoint were responsible for the prolonged metaphase. Live cell imaging showed that kinetochore Mad2 became undetectable soon after chromosome congression. Measurements …

Brain Uptake, Retention, And Efflux Of Aluminum And Manganese, Robert A. Yokel

Pharmaceutical Sciences Faculty Publications

My colleagues and I investigated the sites and mechanisms of aluminum (Al) and manganese (Mn) distribution through the blood-brain barrier (BBB). Microdialysis was used to sample non-protein-bound Al in the extracellular fluid (ECF) of blood (plasma) and brain. Brain ECF Al appearance after intravenous Al citrate injection was too rapid to attribute to diffusion or to transferrin-receptor-mediated endocytosis, suggesting another carrier-mediated process. The brain:blood ECF Al concentration ratio was 0.15 at constant blood and brain ECF Al concentrations, suggesting carrier-mediated brain Al efflux. Pharmacological manipulations suggested the efflux carrier might be a monocarboxylate transporter (MCT). However, the lack of Al …

A 12-Year Prospective Study Of The Long-Term Effects Of Early Child Physical Maltreatment On Psychological, Behavioral, And Academic Problems In Adolescence, Jennifer E Lansford, Kenneth A Dodge, Gregory S Pettit, John E Bates, Joseph Crozier, Julie Kaplow

Faculty, Staff and Students Publications

OBJECTIVE: To determine whether child physical maltreatment early in life has long-term effects on psychological, behavioral, and academic problems independent of other characteristics associated with maltreatment.

DESIGN: Prospective longitudinal study with data collected annually from 1987 through 1999.

SETTING AND PARTICIPANTS: Randomly selected, community-based samples of 585 children from the ongoing Child Development Project were recruited the summer before children entered kindergarten in 3 geographic sites. Seventy-nine percent continued to participate in grade 11. The initial in-home interviews revealed that 69 children (11.8%) had experienced physical maltreatment prior to kindergarten matriculation.

MAIN OUTCOME MEASURES: Adolescent assessment of school grades, standardized …