Showing papers by "University of Münster published in 2003"

From vulnerable plaque to vulnerable patient: a call for new definitions and risk assessment strategies: Part II.

Abstract: Atherosclerotic cardiovascular disease results in >19 million deaths annually, and coronary heart disease accounts for the majority of this toll. Despite major advances in treatment of coronary heart disease patients, a large number of victims of the disease who are apparently healthy die suddenly without prior symptoms. Available screening and diagnostic methods are insufficient to identify the victims before the event occurs. The recognition of the role of the vulnerable plaque has opened new avenues of opportunity in the field of cardiovascular medicine. This consensus document concludes the following. (1) Rupture-prone plaques are not the only vulnerable plaques. All types of atherosclerotic plaques with high likelihood of thrombotic complications and rapid progression should be considered as vulnerable plaques. We propose a classification for clinical as well as pathological evaluation of vulnerable plaques. (2) Vulnerable plaques are not the only culprit factors for the development of acute coronary syndromes, myocardial infarction, and sudden cardiac death. Vulnerable blood (prone to thrombosis) and vulnerable myocardium (prone to fatal arrhythmia) play an important role in the outcome. Therefore, the term "vulnerable patient" may be more appropriate and is proposed now for the identification of subjects with high likelihood of developing cardiac events in the near future. (3) A quantitative method for cumulative risk assessment of vulnerable patients needs to be developed that may include variables based on plaque, blood, and myocardial vulnerability. In Part I of this consensus document, we cover the new definition of vulnerable plaque and its relationship with vulnerable patients. Part II of this consensus document focuses on vulnerable blood and vulnerable myocardium and provide an outline of overall risk assessment of vulnerable patients. Parts I and II are meant to provide a general consensus and overviews the new field of vulnerable patient. Recently developed assays (eg, C-reactive protein), imaging techniques (eg, CT and MRI), noninvasive electrophysiological tests (for vulnerable myocardium), and emerging catheters (to localize and characterize vulnerable plaque) in combination with future genomic and proteomic techniques will guide us in the search for vulnerable patients. It will also lead to the development and deployment of new therapies and ultimately to reduce the incidence of acute coronary syndromes and sudden cardiac death. We encourage healthcare policy makers to promote translational research for screening and treatment of vulnerable patients.

MALAT-1, a novel noncoding RNA, and thymosin beta4 predict metastasis and survival in early-stage non-small cell lung cancer.

Abstract: Early-stage non-small cell lung cancer (NSCLC) can be cured by surgical resection, but a substantial fraction of patients ultimately dies due to distant metastasis. In this study, we used subtractive hybridization to identify gene expression differences in stage I NSCLC tumors that either did or did not metastasize in the course of disease. Individual clones (n=225) were sequenced and quantitative RT-PCR verified overexpression in metastasizing samples. Several of the identified genes (eIF4A1, thymosin beta4 and a novel transcript named MALAT-1) were demonstrated to be significantly associated with metastasis in NSCLC patients (n=70). The genes' association with metastasis was stage- and histology specific. The Kaplan-Meier analyses identified MALAT-1 and thymosin beta4 as prognostic parameters for patient survival in stage I NSCLC. The novel MALAT-1 transcript is a noncoding RNA of more than 8000 nt expressed from chromosome 11q13. It is highly expressed in lung, pancreas and other healthy organs as well as in NSCLC. MALAT-1 expressed sequences are conserved across several species indicating its potentially important function. Taken together, these data contribute to the identification of early-stage NSCLC patients that are at high risk to develop metastasis. The identification of MALAT-1 emphasizes the potential role of noncoding RNAs in human cancer.

Typing of methicillin-resistant Staphylococcus aureus in a university hospital setting by using novel software for spa repeat determination and database management

Abstract: The spa gene of Staphylococcus aureus encodes protein A and is used for typing of methicillin-resistant Staphylococcus aureus (MRSA) We used sequence typing of the spa gene repeat region to study the epidemiology of MRSA at a German university hospital One hundred seven and 84 strains were studied during two periods of 10 and 4 months, respectively Repeats and spa types were determined by Ridom StaphType, a novel software tool allowing rapid repeat determination, data management and retrieval, and Internet-based assignment of new spa types following automatic quality control of DNA sequence chromatograms Isolates representative of the most abundant spa types were subjected to multilocus sequence typing and pulsed-field gel electrophoresis One of two predominant spa types was replaced by a clonally related variant in the second study period Ten unique spa types, which were equally distributed in both study periods, were recovered The data show a rapid dynamics of clone circulation in a university hospital setting spa typing was valuable for tracking of epidemic isolates The data show that disproval of epidemiologically suggested transmissions of MRSA is one of the main objectives of spa typing in departments with a high incidence of MRSA

Proinflammatory Cytokines Disrupt Epithelial Barrier Function by Apoptosis-Independent Mechanisms

Abstract: It is well known that inflammatory conditions of the intestinal mucosa result in compromised barrier function. Inflammation is characterized by an influx into the mucosa of immune cells that influence epithelial function by releasing proinflammatory cytokines such as IFN-γ and TNF-α. Mucosal barrier function is regulated by the epithelial apical junctional complex (AJC) consisting of the tight junction and the adherens junction. Since the AJC regulates barrier function, we analyzed the influence of IFN-γ and TNF-α on its structure/function and determined the contribution of apoptosis to this process using a model intestinal epithelial cell line, T84, and IFN-γ and TNF-α. AJC structure/function was analyzed by confocal microscopy, biochemical analysis, and physiologic measurement of epithelial gate/fence function. Apoptosis was monitored by determining cytokeratin 18 cleavage and caspase-3 activation. IFN-γ induced time-dependent disruptions in epithelial gate function that were potentiated by coincubation with TNF-α. Tight junction fence function was somewhat disrupted. Cytokine treatment was associated with internalization of AJC transmembrane proteins, junction adhesion molecule 1, occludin, and claudin-1/4 with minimal effects on the cytoplasmic plaque protein zonula occludens 1. Detergent solubility profiles of junction adhesion molecule 1 and E-cadherin and their affiliation with “raft-like” membrane microdomains were modified by these cytokines. Inhibition of cytokine-induced apoptosis did not block induced permeability defects; further emphasizing their primary influence on the epithelial AJC structure and barrier function. Our findings for the first time clearly separate the proapoptotic effects of IFN-γ and TNF-α from their abilities to disrupt barrier function.

Polyester synthases: natural catalysts for plastics.

Abstract: Polyhydroxyalkanoates (PHAs) are biopolyesters composed of hydroxy fatty acids, which represent a complex class of storage polyesters. They are synthesized by a wide range of different Gram-positive and Gram-negative bacteria, as well as by some Archaea, and are deposited as insoluble cytoplasmic inclusions. Polyester synthases are the key enzymes of polyester biosynthesis and catalyse the conversion of (R)-hydroxyacyl-CoA thioesters to polyesters with the concomitant release of CoA. These soluble enzymes turn into amphipathic enzymes upon covalent catalysis of polyester-chain formation. A self-assembly process is initiated resulting in the formation of insoluble cytoplasmic inclusions with a phospholipid monolayer and covalently attached polyester synthases at the surface. Surface-attached polyester synthases show a marked increase in enzyme activity. These polyester synthases have only recently been biochemically characterized. An overview of these recent findings is provided. At present, 59 polyester synthase structural genes from 45 different bacteria have been cloned and the nucleotide sequences have been obtained. The multiple alignment of the primary structures of these polyester synthases show an overall identity of 8-96% with only eight strictly conserved amino acid residues. Polyester synthases can been assigned to four classes based on their substrate specificity and subunit composition. The current knowledge on the organization of the polyester synthase genes, and other genes encoding proteins related to PHA metabolism, is compiled. In addition, the primary structures of the 59 PHA synthases are aligned and analysed with respect to highly conserved amino acids, and biochemical features of polyester synthases are described. The proposed catalytic mechanism based on similarities to alpha/beta-hydrolases and mutational analysis is discussed. Different threading algorithms suggest that polyester synthases belong to the alpha/beta-hydrolase superfamily, with a conserved cysteine residue as catalytic nucleophile. This review provides a survey of the known biochemical features of these unique enzymes and their proposed catalytic mechanism.

The desorption process in MALDI.

Abstract: A matrix-assisted laser desorption ionization (MALDI) “event” constitutes a complex process, involving optical and mechanical phenomena as well as thermodynamic and physicochemical processes of phase transition and ionization. A successful MALDI analysis encompasses several crucial steps: sample preparation, excitation of sample and disintegration of the condensed phase, generation and separation of charges and ionization of analyte molecules, and, finally, extraction, separation according to the massto-charge ratio of the ions in the mass spectrometer, and detection. Despite the rapid acceptance of the method in chemistry and biomedicine after its introduction by Karas and Hillenkamp,1-4 the underlying mechanisms have been less well understood for a long time and a more comprehensive picture has only recently begun to emerge. Systematic variation of one or more of the relevant “input” parameters is one of the most straightforward experimental tools to obtain insight into the involved mechanisms, and has consequently been utilized in a large number of fundamental studies. The role of the different relevant irradiation (laser) parameters in the desorption/ionization process, as well as those of the matrix and the preparation protocol, have been addressed in these investigations. These included, for example, the role of the laser wavelength, pulse duration, and laser fluence (laser energy per pulse and unit area) for the laser parameters (addressed in section III), and the type of (co-)crystallization and the matter of the incorporation of analyte molecules into matrix crystals for the material side (section IV). In other work, the dynamical parameters of the expanding MALDI particle “plume” have been addressed (section V): the initial kinetic energies and energy distributions of molecules and ions, and the composition of the plume (ion-to-neutral ratio and ejection of particles and clusters versus the emission of molecular constituents). Several researchers have developed theoretical models for the desorption as well as the ionization * E-mail: dreisew@uni-muenster.de. Fax +49-251-8355121. Tel. +49-251-8356726. Klaus Dreisewerd was born in Beckum, Germany, in 1961. He received his diploma in Physics from the University of Münster in 1990 and his Ph.D. degree in 1995 under the supervision of Franz Hillenkamp. He moved to the Free University of Amsterdam as a post-doc in the molecular neurobiology department at the end of 1994 and returned to the Münster institute in 1997. His main current interests are in the field of MALDI fundamentals and that of MALDI-MS with pulsed infrared lasers (IRMALDI). 395 Chem. Rev. 2003, 103, 395−425

S100B in brain damage and neurodegeneration

Abstract: S100B is a calcium-binding peptide produced mainly by astrocytes that exert paracrine and autocrine effects on neurons and glia. Some knowledge has been acquired from in vitro and in vivo animal experiments to understand S100B's roles in cellular energy metabolism, cytoskeleton modification, cell proliferation, and differentiation. Also, insights have been gained regarding the interaction between S100B and the cerebral immune system, and the regulation of S100B activity through serotonergic transmission. Secreted glial S100B exerts trophic or toxic effects depending on its concentration. At nanomolar concentrations, S100B stimulates neurite outgrowth and enhances survival of neurons during development. In contrast, micromolar levels of extracellular S100B in vitro stimulate the expression of proinflammatory cytokines and induce apoptosis. In animal studies, changes in the cerebral concentration of S100B cause behavioral disturbances and cognitive deficits. In humans, increased S100B has been detected with various clinical conditions. Brain trauma and ischemia is associated with increased S100B concentrations, probably due to the destruction of astrocytes. In neurodegenerative, inflammatory and psychiatric diseases, increased S100B levels may be caused by secreted S100B or release from damaged astrocytes. This review summarizes published findings on S100B regarding human brain damage and neurodegeneration. Findings from in vitro and in vivo animal experiments relevant for human neurodegenerative diseases and brain damage are reviewed together with the results of studies on traumatic, ischemic, and inflammatory brain damage as well as neurodegenerative and psychiatric disorders. Methodological problems are discussed and perspectives for future research are outlined.

Elliptic flow of identified hadrons in [formula presented] collisions at [formula presented]

Abstract: The anisotropy parameter (v(2)), the second harmonic of the azimuthal particle distribution, has been measured with the PHENIX detector in Au+Au collisions at roots(NN)=200 GeV for identified and inclusive charged particle production at central rapidities (eta 2 GeV/c, in marked contrast to the predictions of a hydrodynamical model. A quark-coalescence model is also investigated.

Class 3 semaphorins control vascular morphogenesis by inhibiting integrin function

Abstract: The motility and morphogenesis of endothelial cells is controlled by spatio-temporally regulated activation of integrin adhesion receptors, and integrin activation is stimulated by major determinants of vascular remodelling. In order for endothelial cells to be responsive to changes in activator gradients, the adhesiveness of these cells to the extracellular matrix must be dynamic, and negative regulators of integrins could be required. Here we show that during vascular development and experimental angiogenesis, endothelial cells generate autocrine chemorepulsive signals of class 3 semaphorins (SEMA3 proteins) that localize at nascent adhesive sites in spreading endothelial cells. Disrupting endogenous SEMA3 function in endothelial cells stimulates integrin-mediated adhesion and migration to extracellular matrices, whereas exogenous SEMA3 proteins antagonize integrin activation. Misexpression of dominant negative SEMA3 receptors in chick embryo endothelial cells locks integrins in an active conformation, and severely impairs vascular remodelling. Sema3a null mice show vascular defects as well. Thus during angiogenesis endothelial SEMA3 proteins endow the vascular system with the plasticity required for its reshaping by controlling integrin function.

Help Seeking and Help Design in Interactive Learning Environments

Abstract: Many interactive learning environments (ILEs) offer on-demand help, intended to positively influence learning. Recent studies report evidence that although effective help-seeking behavior in ILEs is related to better learning outcomes, learners are not using help facilities effectively. This selective review (a) examines theoretical perspectives on the role of on-demand help in ILEs, (b) reviews literature on the relations between help seeking and learning in ILEs, and (c) identifies reasons for the lack of effective help use. We review the effect of system-related factors, of student-related factors, and of interactions between these factors. The interaction between metacognitive skills and cognitive factors is important for appropriate help seeking, as are a potentially large space of system-related factors as well as interactions among learner- and system-related factors. We suggest directions for future research.

Suppressed π0 Production at Large Transverse Momentum in Central Au + Au Collisions at √sNN = 200 GeV

Abstract: Transverse momentum spectra of neutral pions in the range 1 < p_T < 10 GeV/c have been measured at mid-rapidity by the PHENIX experiment at RHIC in Au+Au collisions at sqrt(s_NN) = 200 GeV. The pi^0 multiplicity in central reactions is significantly below the yields measured at the same sqrt(s_NN) in peripheral Au+Au and p+p reactions scaled by the number of nucleon-nucleon collisions. For the most central bin, the suppression factor is ~2.5 at p_T = 2 GeV/c and increases to ~4-5 at p_T ~= 4 GeV/c. At larger p_T, the suppression remains constant within errors. The deficit is already apparent in semi-peripheral reactions and increases smoothly with centrality.

PHENIX detector overview

Abstract: The PHENIX detector is designed to perform a broad study of A-A, p-A, and p-p collisions to investigate nuclear matter under extreme conditions A wide variety of probes, sensitive to all timescales, are used to study systematic variations with species and energy as well as to measure the spin structure of the nucleon Designing for the needs of the heavy-ion and polarized-proton programs has produced a detector with unparalleled capabilities PHENIX measures electron and muon pairs, photons, and hadrons with excellent energy and momentum resolution The detector consists of a large number of subsystems that are discussed in other papers in this volume The overall design parameters of the detector are presented (C) 2002 Elsevier Science BV All rights reserved

Systematic Investigation of Modern Quantum Chemical Methods to Predict Electronic Circular Dichroism Spectra

Abstract: The ability of different quantum chemical methods to predict experimental electronic circular dichroism (CD) spectra is critically evaluated. Two single-reference, time-dependent approaches based either on density functional theory (TDDFT) or a simplified coupled-cluster expansion (CC2) and two multireference methods (MRMP2 and DFT/MRCI) are considered. The methods are applied to a test suite of seven molecules including a wide range of difficult chromophores (“real-life” examples) and to three model systemsH2S2, twisted ethylene, and dimethyloxiranewhere accurate ab initio MRCI reference data are used for comparison. To investigate the effect of “exact” exchange mixing systematically, the TDDFT calculations were carried out with the BP86, B3-LYP, and BH-LYP functionals. The time-dependent Hartree−Fock (TDHF) method was included as an “upper limit” for the HF-exchange part in the functional. In general, it is found that the accuracy of most of the simulated spectra (except those from TDHF) is good enough ...

Empirical Evidence on Corporate Governance in Europe. The Effect on Stock Returns, Firm Value and Performance

Abstract: In this paper we analyze whether good corporate governance leads to higher common stock returns and enhances firm value in Europe. Throughout this study we use Deminor Corporate Governance Ratings for companies included in the FTSE Eurotop 300. Following the approach of Gompers, Ishii and Metrick (2003) we build portfolios consisting of well-governed and poorly governed companies and compare their performance. We also examine the impact of corporate governance on firm valuation. Our results show a positive relationship between these variables and corporate governance. This relationship weakens substantially after adjusting for country differences. Finally, we analyze the relationship between corporate governance and firm performance, as approximated by Net-Profit-Margin (NPM) and Return-on-Equity (ROE). Surprisingly, and contrary to Gompers, Ishii and Metrick (2003), we find a negative relationship between governance standards and these earnings based performance ratios for which we discuss possible implications.

Re: Leptin—A Growth Factor in Normal and Malignant Breast Cells and for Normal Mammary Gland Development

Abstract: BACKGROUND Obesity is a risk factor for breast cancer in postmenopausal women. As body weight and fat mass increase, circulating leptin increases. Leptin is an adipocyte-derived cytokine that acts through the long form of its receptor, termed OB-Rb. To investigate whether leptin is associated with breast cancer, we determined the expression of OB-Rb in human breast epithelial HBL100 cells and human breast carcinoma-derived T-47D cells, determined whether leptin influenced the proliferation of these cells, and evaluated the structure of mammary tissue in genetically obese leptin-deficient Lep(ob)Lep(ob) and leptin receptor-deficient Lepr(db)Lepr(db) mice. METHODS Cell numbers and cell colony formation by HBL100 and T-47D cells were determined by anchorage-dependent and anchorage-independent growth assays. OB-Rb expression was examined by reverse transcription-polymerase chain reaction and immunoblot analyses. Expression of leptin signaling pathway components was evaluated with immunoblot and electrophoretic mobility shift assays. Mammary gland development in lean and obese mice was investigated in whole-mount studies. All statistical tests were two-sided. RESULTS Leptin enhanced anchorage-dependent proliferation by 138% (95% confidence interval [CI] = 108% to 169%) in T-47D cells and 50% (95% CI = 38% to 60%) in HBL100 cells. In both cell lines, OB-Rb was expressed, and leptin increased the expression of phosphorylated signal transducers and activators of transcription 3 (STAT3), phosphorylated extracellular signal-regulated kinase (ERK), and transcript activator protein 1 (AP-1). However, leptin increased anchorage-independent cell growth only in the breast cancer cell line (by 81% [95% CI = 62% to 101%] compared with untreated cells). Obese Lep(ob)Lep(ob) and Lepr(db)Lepr(db) mice had minimal epithelial development in the mature mammary gland compared with their lean counterparts. CONCLUSIONS Leptin appears to be able to control the proliferation of both normal and malignant breast epithelial cells. Consequently, the leptin pathway should be further studied as a target for interventions to treat or prevent breast cancer.

Zircons from Syros, Cyclades, Greece—Recrystallization and Mobilization of Zircon During High-Pressure Metamorphism

Abstract: Zircons were studied from high-pressure/low-temperature metamorphosed meta-igneous lithologies from Syros. These rocks carry several zircon generations related to each other by dissolution---reprecipitation processes. One generation is pristine zircon that shows growth zoning, relatively elevated contents of trivalent cations and high Th/U ratios. The other end-member is a skeletal zircon generation with negligible trivalent cation contents and low Th/U ratios ( 0 1). Texturally between these two, there is a range of zircon crystals with complex inclusion populations of Y---HREE---Th phases and fluid inclusions, showing variable progress of replacement--recrystallization. Both end-members yield distinct sensitive high-resolution ion microprobe (SHRIMP) U---Pb ages. The pristine generation has an age of 80 2 1 6Ma from a metagabbro, and 76 4 2 1Ma from a meta-plagiogranite dyke. The skeletal, low-Th/U zircon generation yields an age of 52 4 0 8Ma. The older, Late Cretaceous, zircons are interpreted to date emplacement of the magmatic protoliths in a small segment of oceanic crust. The younger, Eocene, age, however, dates a zircon recrystallization event, which possibly coincides with high solubility and mobility of high field strength elements in a high-pressure aqueous fluid phase. Intergrowth relations between zircon and peak-metamorphic garnet, and excellent agreement of the U---Pb ages with white mica Ar---Ar ages for the same samples support the conclusion that Eocene is the true age of high-pressure metamorphism on Syros.

Prevalence of left ventricular diastolic dysfunction in the community. Results from a Doppler echocardiographic-based survey of a population sample.

Abstract: Aims The prevalence of left ventricular diastolic abnormalities in the general population is largely unclear. Thus, the aim of this study was, firstly, to identify abnormal diastolic function by echocardiography in an age-stratified population-based European sample (MONICA Augsburg, \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(n=1274\) \end{document}, 25 to 75 years, mean 51±14) and, secondly, to analyse clinical and anthropometric parameters associated with diastolic abnormalities. Methods and results The overall prevalence of diastolic abnormalities, as defined by the European Study Group on Diastolic Heart Failure (i.e. age dependent isovolumic relaxation time (92–105ms) and early (E-wave) and late (A-wave) left ventricular filling (E/A-ratio, 1–0.5)) was 11.1%. When only subjects treated with diuretics or with left atrial enlargement were considered (suggesting diastolic dysfunction) the prevalence was 3.1%. The prevalence of diastolic abnormalities varied according to age: from 2.8% in individuals aged 25–35 years to 15.8% among those older than 65 years \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \((P{<}0.01)\) \end{document}. Significantly higher rates of diastolic abnormalities were observed in men as compared to women (13.8% vs 8.6%, \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(P{<}0.01\) \end{document}). Independent predictors of diastolic abnormalities were arterial hypertension, evidence of left ventricular (LV) hypertrophy, and coronary artery disease. Interestingly, in the absence of these predisposing conditions, diastolic abnormalities (4.3%) or diastolic dysfunction (1.1%) were rare, even in subjects older than 50 years of age (4.6%) and (1.2%), respectively. In addition to these factors, diastolic dysfunction was related to high body mass index, high body fat mass, and diabetes mellitus. Conclusion The prevalences of diastolic abnormalities and diastolic dysfunction are higher than that of systolic dysfunction and are increased (despite age-dependent diagnostic criteria) in the elderly. However, in the absence of risk factors for diastolic abnormalities or diastolic dysfunction, namely LV hypertrophy, arterial hypertension, coronary artery disease, obesity and diabetes the condition is rare even in elderly subjects. These data allow speculation on whether diastolic heart failure may be prevented by improved implementation of measures directed against predisposing conditions.

Absence of suppression in particle production at large transverse momentum in √sNN = 200 GeV d + Au collisions

Abstract: Transverse momentum spectra of charged hadrons with p(T)<8 GeV/c and neutral pions with p(T)<10 GeV/c have been measured at midrapidity by the PHENIX experiment at BNL RHIC in d+Au collisions at sqrt[s(NN)]=200 GeV. The measured yields are compared to those in p+p collisions at the same sqrt[s(NN)] scaled up by the number of underlying nucleon-nucleon collisions in d+Au. The yield ratio does not show the suppression observed in central Au+Au collisions at RHIC. Instead, there is a small enhancement in the yield of high momentum particles.

National Identity in a United Germany: Nationalism or Patriotism? An Empirical Test With Representative Data

Abstract: Nationalism and patriotism can be thought of as consequences of national identity that represent positive evaluations of one's own group but imply different social goals. This paper investigates the ways in which these concepts are related to attitudes toward minorities. The data analyzed were drawn from a representative sample of residents of the former East and West Germany who responded to items on the national identity of Germans in 1996 as part of a panel study. A model with multiple indicators was tested via a multiple-group analysis of a structural equations model followed by latent class analyses. Both East and West Germans displayed attitudinal patterns that link national identity with tolerance toward others; in both subsamples, nationalism and patriotism were respectively associated with greater intolerance and greater tolerance toward minorities.