Dynamic changes in heparan sulfate during muscle differentiation and ageing regulate myoblast cell fate and FGF2 signalling

Satellite cells (SCs) are skeletal muscle stem cells residing quiescent around healthy muscle fibres. In response to injury or disease SCs activate, proliferate and eventually differentiate and fuse to one another to form new muscle fibres, or to existing damaged fibres to repair them. The sulfated polysaccharide heparan sulfate (HS) is a highly variable biomolecule known to play key roles in the regulation of cell fate decisions, though the changes that muscle HS undergoes during SC differentiation are unknown. Here we show that the sulfation levels of HS increase during SC differentiation; more specifically, we observe an increase in 6-O and 2-O-sulfation in N-acetylated disaccharides. Interestingly, a specific increase in 6-O sulfation is also observed in the heparanome of ageing muscle, which we show leads to promotion of FGF2 signalling and satellite cell proliferation, suggesting a role for the heparanome dynamics in age-associated loss of quiescence. Addition of HS mimetics to differentiating SC cultures results in differential effects: an oversulfated HS mimetic increases differentiation and inhibits FGF2 signalling, a known major promoter of SC proliferation and inhibitor of differentiation. In contrast, FGF2 signalling is promoted by an N-acetylated HS mimetic, which inhibits differentiation and promotes SC expansion. We conclude that the heparanome of SCs is dynamically regulated during muscle differentiation and ageing, and that such changes might account for some of the phenotypes and signalling events that are associated with these processes

Heparan sulfate phage display antibodies recognise epitopes defined by a combination of sugar sequence and cation binding.

Phage display antibodies are widely used to follow heparan sulfate (HS) expression in tissues and cells. We demonstrate by ELISA, that cations alter phage display antibody binding profiles to HS and this is mediated by changes in polysaccharide conformation, demonstrated by circular dichroism spectroscopy. Native HS structures, expressed on the cell surfaces of neuroblastoma and fibroblast cells, also exhibited altered antibody binding profiles following exposure to low mM concentrations of these cations. Phage display antibodies recognise conformationally-defined HS epitopes, rather than sequence alone, as has been assumed, and resemble proteins in being sensitive to changes in both charge distribution and conformation following binding of cations to HS polysaccharides

Heparin Isomeric Oligosaccharide Separation Using Volatile Salt Strong Anion Exchange Chromatography

The complexity of heparin and heparan sulfate saccharides makes their purification, including many isomeric structures, very challenging and is a bottleneck for structure–activity studies. High-resolution separations have been achieved by strong anion exchange (SAX) chromatography on Propac PA1 and cetyltrimethylammonium (CTA)-C18 silica columns; however, these entail subsequent desalting methodologies and consequent sample losses and are incompatible with orthogonal chromatography methodologies and, in particular, mass spectrometry. Here, we present the CTA-SAX purification of heparin oligosaccharides using volatile salt (VS) buffer. In VSCTA-SAX, the use of ammonium bicarbonate buffer for elution improves resolution through both weaker dissociation and conformational coordination of the ammonium across the sulfate groups. Using ion mobility mass spectrometry, we demonstrate that isomeric structures have different structural conformations, which makes chromatographic separation achievable. Resolution of such structures is improved compared to standard SAX methods, and in addition, VSCTA-SAX provides an orthogonal method to isolate saccharides with higher purity. Because ammonium bicarbonate is used, the samples can be evaporated rather than desalted, preventing substantial sample loss and allowing more effective subsequent analysis by electrospray mass spectrometry. We conclude that VSCTA-SAX is a powerful new tool that helps address the difficult challenge of heparin/heparan sulfate saccharide separation and will enhance structure–activity studies

Intricate Correlation between Body Posture, Personality Trait and Incidence of Body Pain: A Cross-Referential Study Report

OBJECTIVE: Occupational back pain is a disorder that commonly affects the working population, resulting in disability, health-care utilization, and a heavy socioeconomic burden. Although the etiology of occupational pain remains largely unsolved, anecdotal evidence exists for the contribution of personality and posture to long-term pain management, pointing to a direct contribution of the mind-body axis. In the current study, we have conducted an extensive evaluation into the relationships between posture and personality. METHOD: We have sampled a random population of 100 subjects (50 men and 50 women) in the age range of 13-82 years based on their personality and biomechanical profiles. All subjects were French-Canadian, living in Canada between the Québec and Sorel-Tracy areas. The Biotonix analyses and report were used on the subjects being tested in order to distinguish postural deviations. Personality was determined by using the Myers-Briggs Type Indicator questionnaire. RESULTS: We establish a correlation between ideal and kyphosis-lordosis postures and extraverted personalities. Conversely, our studies establish a correlative relationship between flat back and sway-back postures with introverted personalities. CONCLUSION: Overall, our studies establish a novel correlative relationship between personality, posture and pain

Heparin and Heparan Sulfate: Analyzing Structure and Microheterogeneity [chapter]

available in PMC 2013 August 28The structural microheterogeneity of heparin and heparan sulfate is one of the major reasons for the multifunctionality exhibited by this class of molecules. In a physiological context, these molecules primarily exert their effects extracellularly by mediating key processes of cellular cross-talk and signaling leading to the modulation of a number of different biological activities including development, cell proliferation, and inflammation. This structural diversity is biosynthetically imprinted in a nontemplate-driven manner and may also be dynamically remodeled as cellular function changes. Understanding the structural information encoded in these molecules forms the basis for attempting to understand the complex biology they mediate. This chapter provides an overview of the origin of the structural microheterogeneity observed in heparin and heparan sulfate, and the orthogonal analytical methodologies that are required to help decipher this information

FACES -TUNNEKORTIT : apuna tunteiden käsittelyssä ja ilmaisussa

Tässä toiminnallisessa opinnäytetyössä kehitettiin kriisitilanteisiin sopivat tunnekortit. Idea tunnekorttien tekemiseen lähti työmme toimeksiantajalta. Työn toimeksiantaja on Ylä-Savon SOTE -kuntayhtymän hyvinvointipalveluiden perhekeskukseen kuuluvassa perheneuvolan koulutiimissä työskentelvä vastaava kuraattori. Hän työskentelee Iisalmessa ala- ja yläkouluissa sekä lyseossa. Hän kaipasi keskustelun tueksi työvälinettä, jonka avulla lasten ja nuorten elämää koskettavista vaikeista asioista olisi helpompi puhua. Vaikeista asioista puhuminen on tarpeellista, jotta asiat eivät jää käsittelemättä. Kehittämistyön tuloksena syntyi yhteensä 17 tunnekorttia. Opinnäytetyön teoreettisessa viitekehyksessä käsitellään ennaltaehkäisevää työtä koulussa, lapsen tunne-elämän kehitystä sekä kouluikäisten lasten ja nuorten kriisejä ja surua. Kriiseistä selviytymisen tukeminen on myös yksi keskeisimmistä sisällöistä viitekehyksessä. Opinnäytetyön loppuosassa kuvataan opinnäytetyöprosessin ja tunne-korttien kehittämisprosessin vaiheita. Taide on vahva ilmaisun keino ja se herättää paljon ajatuksia ja tunteita. Halusimme tuoda esiiin omaa taiteellista osaamistamme, joten valitsimme korttien tekotavaksi piirtämisen. Piirsimme kortteihin ihmisen kasvoja, joissa näkyy erilaisia tunnetiloja. Tunnetiloja on kuvattu erilaisten ilmeiden ja eleiden avulla. Kortit ovat mustavalkoisia. Tunnekorttien perusajatus on sanoittaa tunteita. Tunnekortteja on olemassa jo monenlaisia, mutta erityisesti kriisi-tilanteisiin sopivia kortteja ei ole kuin muutamia markkinoilla. Toimeksiantaja käytti tunnekortteja 8-17 vuotiaiden lasten ja nuorten kanssa sekä yksilö- että ryhmätapaamisissa. Testijakso kesti noin 10 viikkoa. Testitulosten perusteella olemme onnistuneet kehittämään toimivan työvälineen, joka hyödyttää sekä toimeksiantajaa että meitä itseämme tulevina sosiaalialan ammattilaisina. Korttien soveltamismahdollisuudet erilaisissa ryhmä- ja yksilötilanteissa ovat laajat. Kortteja voi käyttää myös muissa toimintaym-päristöissä kuin kouluissa. Jatkamme tunnekorttien kehittämistä saatujen kehittämisehdotusten pohjalta vielä tämän opinnäytetyöprosessin jälkeen.The target of this functional thesis was to develop emotion cards which can be used with school aged children and adolescents when dealing with different kinds of crises in everyday life. The client organization in this thesis was Ylä-Savon SOTE Joint Municipal Authority in Social and Health Care and the person who ordered these cards was a school social worker in Iisalmi. The school social worker told that she needed some kind of a tool by which it woud be easier to talk about life’s challenges and difficulties with children and adolescents. It is important and necessary to talk about life’s challenges and difficulties because otherwise it could cause mental health problems later on. It also makes a person feel better if he or she can talk about the difficult issues with someone. During this development process 17 emotion cards were created. The theory part of this thesis handles preventive work in school, child’s emotional development and also crises and sorrow in school aged childrens’ lives. One of the most important things is to support the child who has gone through some kind of a crisis. This is also one of the main points in the theory part of this thesis. The last chapters of this thesis are describing the process of this thesis and how the emotion cards were made. The fine arts are a very powerful way of expression. They also rouse a lot of thoughts and feelings. In this thesis we wanted to use our own talents and that is why we chose to draw the pictures of the emotion cards by ourselves. On the emotion cards there are pictures of human faces with different kinds of facial expressions. The emotion cards are greyscaled. The main point of the emotion cards is to help to express feelings and to talk about them. There are a lot of different kinds of emotion card series on sale but just a few which are designed especially to help to cope with the crises. The school social worker used the emotion cards with children and adolescents aged 8 to 17 year-olds in personal meetings and in group sessions. The experimentation period of cards lasted 10 weeks. According to the results of experimentation a tool was created which is helping the school social worker to bring up life’s difficulties and crises which children and adolescents are facing in everyday life. The emotion cards can be used in many different ways and also in different kinds of surroundings in social work. The emotion cards will be still improved and finished after the given improvement ideas after the process of this thesis

Chemical synthesis of a sulfated d-glucosamine library and evaluation of cell proliferation capabilities.

'No abstract

Single-entity heparan sulfate glycomimetic clusters for therapeutic applications

Heparan sulfate (HS) is a highly sulfated glycosaminoglycan with a variety of critical functions in cell signaling and regulation. HS oligosaccharides can mimic or interfere with HS functions in biological systems; however, their exploitation has been hindered by the complexity of their synthesis. Polyvalent displays of small specific HS structures on dendritic cores offer more accessible constructs with potential advantages as therapeutics, but the synthesis of single-entity HS polyvalent compounds has not previously been described. Herein we report the synthesis of a novel targeted library of single-entity glycomimetic clusters capped with varied HS saccharides. They have the ability to mimic longer natural HS saccharides in their inhibition of the Alzheimer's disease (AD) protease BACE-1. We have identified several single-entity HS clusters with IC50 values in the low-nanomolar range. These HS clusters are drug leads for AD and offer a novel framework for the manipulation of heparan sulfate-protein interactions in general