November 21, 2018

2019 Call to research Grant ASI

350,000 EURO FOR RESEARCH ON ANGELMAN SYNDROME

Call for Applications for Research Grants into Angelman Syndrome

Deadline 4th January 2019

Angelman Syndrome is a rare neurogenetic disorder, resulting from the loss of function of a single gene, UBE3A, which encodes for the ubiquitin ligase E6-AP. It is characterised by severe intellectual disability, lack of speech, motor coordination deficits, sleep disturbance and an unusually happy demeanour.

The international Angelman Syndrome Alliance (ASA) is an initiative of parent support organisations from 15 countries (including Ireland, UK, Belgium, Netherlands, Germany, Austria, France, Italy, Portugal, Japan, Israel, and Spain) who have pooled resources and funds in order to advance research into the syndrome.

The ASA is now inviting applications from researchers from any country for grants of up to €175,000 (total grant fund €350,000). Proposals covering preclinical (basic) research as well as translational research are welcomed, however applications for behavioural therapies are not eligible at this point.

The duration of the grant is flexible up to a maximum of 4 years. After 50% completion of the anticipated duration, an evaluation of the project’s progress will be expected. This will be in the form of both a detailed written scientific report for the Scientific Advisory Board (SAB) and an update in more general layman’s terms to share with the parent organisations who fund the grant.

This is a two-stage application process. In the first instance, a short initial application should be submitted via the ASA website (www.angelmanalliance.org) by 12:00 noon on 4th January 2019 and should include the following:

  1. A cover letter, including the title of the proposal and the name of the principal investigator.
  2. A two-page summary of the proposed research including paragraphs on the hypothesis, background, research plan, principal methodology, and the significance of the proposed research. An overview of the budget must also be included (indirect costs cannot be applied for).
  3. The Curriculum vitae of the applicant.

All applications should be written in English, in Times New Roman, 10 point, 1.5 spaced on A4 paper.

From 14th January 2019, detailed applications will be invited for proposals passing stage 1, to be submitted by 12:00 noon on or before 11th March 2019. Successful applicants will be informed by 22nd July 2019 and will be invited to give the ASA-AWARD LECTURE at the next biennial ASA scientific conference in autumn 2020.

Proposals involving laboratory animal testing must be approved by the respective institutional review boards before funding can be released.  Research applications from outside the EU, must still conform to ethical guidelines and laws within the EU.

à Questions about the grant process should be directed to the ASA Board at: Manuel Duarte – direccao@angel.pt

à Questions about research on Angelman Syndrome should be directed to the Scientific Advisory Board at: Harald Sitte –  harald.sitte@meduniwien.ac.at

 

www.angelman.ie

 

October 25, 2018

Research updates and summary Germany 2018

See below  the latest summary of Alliance Funded research presented in Germany October 2018. Thanks to Annette Kent who has summarised it in a family friendly version.

 

Summary report from the 6th International Scientific Conference, Hamburg, Germany by Annette Kent Bsc PhD, on behalf of Angelman Syndrome Ireland

Molecular mechanisms underlying Angelman Syndrome (AS) and investigating treatment options 

Rosella Avagliano Trezza, Centre for Neurodevelopmental Disorders, Dept of Neurosciences, Erasmus MC, Netherlands

This work aimed to understand the key proteins in the cytoplasm and nucleus of the cell, exploring the circumstances under which the various forms of Ube3a move or are retained in either location.  This work showed that 80% of the ube3a is the short form and 20% is the long form.

Removing the AZUL domain changed the location of the short form from nucleus to the cytoplasm and it is thought that the AZUL domain is necessary but not sufficient for Ube3a nuclear localization.

Doing experiments using leptomycin (inhibitor of movement from the nucleus) showed that the short form starts and stays nuclear however the short form without the Azul domain stays in the cytoplasm.

*AZUL domain is a specific location on Uba3e gene.

*Individual cells are comprised of a nucleus (the part of the cell that contains the genetic material) and the cytoplasm.  The cytoplasm describes all the material in the cells outside of the nucleus.
A behavioural test battery for mouse models for AS: a powerful tool for testing drugs and novel Ube3a mutants:

Monica Sonzogoni, Centre for Neurodevelopmental Disorders, Dept of Neurosciences, Erasmus MC, Netherlands

This group previously described the characteristics/phenotypes of AS, and their work now focused on testing the robustness of these characteristics using a battery of behaviour tests.  It was pointed out that there are important clinical questions to focus on including: information on meaningful outcome measures, assessment of the characteristics and various interventions.

A conclusion from this work was that the AS mouse model can be used to assess the majority of the shared AS characteristics except the absence of speech, providing confirmation that AS characteristics can be identified, can be put into a mouse model and can be used for drug testing.
Cellular phenotypes of human AS iPSC derived neurons

Stormy Chamberlain, Dept of Genetics and Genome Sciences, UConn Health, USA

This work focused on stem cell derived human neurons and characterisation of neurons generated from people with AS.  It aimed to understand neuronal maturation and seeing whether specific phenotypes could be made from human AS neurons. One of the key reasons for doing this work was to help develop future therapies.

The generated neurons facilitated various studies, used to assess whether specific mutations/changes in Ube3a are likely to cause AS and whether therapeutic approaches can successfully restore neuronal function.

*iPSC – induced Pluripotent Stem cell culture are those generated from adult cells.  Pluripotent stem cell are master cells. They can potentially produce any cell or tissue the body needs to repair itself. This “master” property is called pluripotency.

*Phenotype is the set of observable characteristics of an individual resulting from the interactions of its genes or genotype with the environment.

 

 

Potential therapeutic approach for synaptic deficit in AS – the JNK inhibitor peptide (human in vitro model)

Tiziano Borsello, IRCCS Cytogenetics and Molecular Genetics Lab, Istito Auxologico Italiano, Milano, Italy

This work focused on the role of JNK at the nerve synapse.  It is known that Ube3a loss impairs synaptic transmission but the mechanism involved needs to be further understood.  Early data from this work shows that differences exist between samples from people with AS and controls (i.e. non-AS).   JNK plays a significant role and works by controlling a number of cellular processes including proliferation, embryonic development and apoptosis. It is thought that differences observed in AS group between the males and females may be due to hormonal differences.

*Apoptosis is the death of cells as a normal and controlled part of development and growth.

*JNK, c-jun N terminal kinase, a group of enzymes which play an important an important role in the regulation of neuronal functions. *

*Synapses is a structure that allows a nerve cell to pass an electrical or chemical signal to another nerve.
AS treatment opportunities and clinical biomarkers

Ben Philpot, University of North Carolina, USA

This work focused on ‘Epiloptogenesis’ which is the serialisation of seizures i.e. the gradual process of becoming more susceptible to seizures.  Using mice, they showed that the loss of Ube3a from GABAergic neurons (inhibitory neurons) increased seizure sensitivity which was not observed in glutamerigic neurons (excitatory neurons).

Tests carried out using mice showed similar susceptibility in early stages of the experiments, with larger differences observed in the later stages in the normal vs the AS mice (greater susceptibility in AS neurons).  They also used a hyperthermia induced seizure model where it was observed that seizures occur at much lower temperatures in the AS mice than in normal mice.

A very interesting piece of work gave the results of deleting ube3a in excitatory and inhibitory neurons followed by their reinstatement, helping to understand the reversibility of the epiloptogenesis. Reinstatement of ube3a reduced susceptibility in juvenile mice which was not seen in adult AS.

 

 

GABA mediated inhibitory dysfunction and its pathophysiological contribution in AS

 Kiyoshi Egawa, Pediatric Dept., Hokkaido University Graduate School of Medicine, Japan

Analysis of the somatosensory evoked response using magnetoencephalography was performed and shown to be modified by GABAa receptor agonists. The total amount of GABAa receptor is not decreased in people AS. Tonic inhibition is different in different parts of the brain and maybe due to the GAT1 (GABA transporter 1).  Dysregulation of tonic inhibition is a potential target for developing pharmacological strategy for AS.

*Tonic inhibition is a constant, long-lasting inhibition which is in contrast with phasic inhibition describing short intermittent bursts of inhibition.  Recently, gain, rather than loss, of tonic inhibition has been shown to underlie the pathophysiology of epilepsy.

*Somatosensory is a part of the sensory nervous system which is made up a sensory neurons and pathways.
Cannabidiol extract as a new approach in the anticonvulsive treatment of the Angelman Syndrome – based on neurotransmitter profile, particularly in consideration of GABA and glutamate

Christel KannegieBer-Leitner, Research Group Angelman e.V., Germany

This presentation focused on the use cannabidiol extract (CBD) in AS a potential new approach.  Details of experience with a young man, the son of Christel, with AS and NCSE (Non-Convulsive Status Epilepticus) who had immediate freedom from seizures for 8 months after treatment with CBD.

A stronger glutamate was found in Magnesium (Osborn et al., The Journal of Clinical Pharmacology, 2016, 56(3) 260-265).  The experience was that irregular use of Magnesium could be useful.  However, questions remain on both these approaches and research is needed to understand long- and short-term side and also the role that genetics may play.
Imprinting defects and differential diagnosis

Bernhard Horsthemke, Jasmin Beygo and Karin Buiting Universitatsklinikum, Essen, Germany

This presentation focused on the imprinting defects and differential diagnoses within the family of AS syndromes.
Adeno Associated Virus Mediated Ube3a gene or protein replacement strategies

Ed Weber, Molecular Pharmacology and Physiology, CSO Byrd Alzheimer’s Institute, USA and also for AGILIS Biotherapeutics, USA

This talk focused on a gene therapy shown to restore learning and memory in a mouse model. This group previously showed a recovery in spatial and associated memory defects as well as the underlying alteration in hippocampal synaptic plasticity following the use of “Adeno Associated Virus” also known as AAV to deliver either the ube3a protein or gene.

This work is advancing and the current focus is on bioavailability. The phase I clinical trial protocol is being worked on however it needs good outcome measures but plans to be active for late 2020.

*Bioavailability is the proportion of a drug or other substance which enters the circulation when given to an individual and able to have effect.
Potential therapeutic approach for synaptic deficit in AS: the JUNK peptide (mouse model in vivo)

Silvia Russo, Neuronal Death and Neuroprotection Lab, Dept of Pharmacological and Biomolecular Sciences, CEND, Milano, Italy

Potential therapeutic approach for synaptic deficit in AS with the aim of creating a patient specific model to detect biomarkers (morphological biochemical ad functional biomarkers).
Natural history study efforts in the AS and the scientific rationale for therapy with antisense oligonucleotides

Rebecca Crean, Director, Clinical Development, IONIS, USA

IONIS are working on anitsense oligonucleotide (ASO) therapy as a platform and focused on AS to unsilence a protein. IONIS are working on a large number of ASOs for activity in multiple tissues.  ASOs do not cross the blood brain brain so administration is via bolus intra-thecal injections. In AS paternal ube3a is silenced, using an ASO this could be unsilenced and therefore could be a novel approach to treat AS.

Foundational science needs to enable feasible efficient drug development and assist in the understanding the impact on the individual patient measures including patient reported outcomes will be very important.

IONIS is focused on developing natural history studies which will facilitate drug development.  Natural history studies are studies on individuals over time who have or are at risk of developing a specific medical condition. They allow door greater understanding of the disease and help guide clinical trials.  The main gaps in knowledge are longitudinal changes, biomarkers, correlation between ube3a levels and clinical features.  A number of international registries exist which assist including the following, Backpackhealth.com, AS registry, China Angelman registry, and Angelman Biomarkers and Outcomes Measures (ABOM).

*Anti-sense oligonucleotides are short, synthetic, single-stranded DNA (that can alter protein production.

*Intra-thecal is a route of administration for drugs via an injection into spinal cord
OVID Therapeutics: Update on the AS STARS: Topline results from a phase 2 adults and adolescent AS clinical trial.  A randomised, double blind, safety and efficacy study of Gaboxadol (OV101)

Amit Rakhit, Chief Medical and Portfolio Officer, OVID Therapeutics, USA.

This presentation focused on the development work that OVID are focused on in particular the STARS clinical trial using OV101.   OV101 is a selective GABAa receptor agonists which was tested in this phase II study in 3 groups I.e. placebo, once daily dosing and twice daily dosing (10mg, 15mg).

A total of 88 peope with AS were recruited to the study and assessments completed included were adverse events, global symptoms, behaviour scales and sleep diaries.

The primary endpoint of the study was met in that the agent was safe and tolerable as measured by the adverse events.  Some exploratory end points were assessed at 12 weeks with global functioning AS assessed by a Clinician Global Impression-improvement (CGI-I) was statistically different between placebo and the treatment groups, with a 27.4% difference being reported.

It is recognised in their studies that behaviour scales need to be altered to consider non-verbal nature of AS.  Additional data is presented in 2 weeks at the international conference for American Academy of Child and Adolescent Psychiatry.
December 20, 2017

Angelman Syndrome Alliance Research update

ASI as part of our ongoing funding of  research are delighted to be able to provide a summary of the projects to date. Kindly complied by Annette Kent, they are explained in a family friendly version with a glossary to assist. Please see our research page on this website for the full summary.

 

November 25, 2016

Angelman European Alliance

We were delighted to be represented at the 2016 International Scientific Conference by Sarah Roarty and Annette Kent. The following is a summary of the two awardees, whom we are contributing funding to. It also explains the 2016 awardees. A glossary of terms at the end is also included to help as background. This was a really well attended meeting of parents, scientists and country organizations from many parts of the world, making it a truly international meeting.

SUMMARY:

Stormy Chamberlain, Assistant Professor of Genetics and Genome Sciences, University of Connecticut Health Centre, kicked off what was a day of high level science. Stormy spoke about her work with pluripotent stem cells and the steps they are taking to generate and identify an Angelman Syndrome (AS) neurons generated from these stem cells. Once generated these AS neurons will facilitate greater understanding of events in AS.

Ben Philpot, Department of Cell Biology and Physiology, University of North Carolina, later in the day gave a very clear concise summary of his focus on the white matter deficits in AS and the meaning of the same. He also spoke about the hyper-excitability within AS and the loss of UBE3A from inhibitory neurons increasing the susceptibility to seizures.

Chloe Simmons from FAST Australia, presented on the Global Registry that has been established by FAAT Australia for AS. This registry aims to capture information about individual AS patients to provide a collection of data to assist with future research.

Other presentations included details on the key features of AS mouse model, genetic approaches to reinstatement of the UBE3A, and novel therapeutic approaches. There were presentations also from the awardees from previous ASA funds and more recent awardees. Summaries of this work is given below:

Previous awardees
1. Dr. Ben Distal, AMC Amsterdam
Target identification for E6AP

The purpose of this project was to identify the target for E6AP, the protein produced from the UBE3A gene. This work also focused on the breakdown pathway involving proteasomes and the ubiquitination process, a process by which very small molecules are tagged with ubiquitin molecules and degraded by proteasomes.

Experiments which included proteasome quantification and in-vitro system bacterial ubiquitination, confirmed the presence of 4 interactors. In particular, there was the RPN10 molecules that binds to E6AP, the AZUL domain. Identifying the target of E6AP enables understanding of how the protein molecules work and what action could be taken.

2. Dr. Geeske van Woerden – Prof. Dr. Ype Elgersma
In vivo characterization of proteins that interact with UBE3A.

This work relates to the characterization of proteins, PML and RPN10 which are overexpressed in in-vitro systems when UBE3A is deficient. In-vitro experiments on cells have studied the alterations in neuron shape. In-vivo experiments using in-utero mice have used the electroporation (a process of introducing DNA or chromosomes into bacteria or other cells using an electrical pulse to open the pores in the cell membranes briefly) method and observed changes in the neuronal migration and UBE3A silencing observed a neuronal migration deficit in the developing mice.

PML – higher expression of the PML gene was observed in the mutant mice (PML gene encodes a protein which is a tumor suppressor, promyelocytic leukemia). PML has been shown to play an important role in the development of the neocortex. In-vitro experiments show that the overexpression of PML impairs the maturation of neurons, the in-vivo equivalent has shown that the increased levels of PML results in profound defects of migration, in that reducing the levels of this protein shows an improvement on cognitive and motor tests in animal model of AS.

RPN10 – It shows that RPN10 is a substrate for almost all E3 ligases, it is not degraded by UBE3A. Reduction in RPN10 can result in a migration delay, while the overexpression has no effect. Since it is an important protein for brain development, work is still ongoing to figure out what its function in AS.
2016 awardees

1. Dr. ssa Silvia Russo – Dr. ssa Tizana Borsella
Characterisation of synapses of neurons in AS

Recent studies have shown that cognitive deficits in AS are partly due to defects in the synapses of neurons of patients. In these special structures, called synapses, they are irregular shape, there are fewer of them and they work differently. Synapses are critical for learning memory and cognitive function, as they represent the point of contact between neurons and are necessary for the passage of information within neurons of our brain.

They will study the mechanisms that causes the changes in synapses in AS in an animal model (mice) and a human model. The Russian partners will generate human models using stem cells from patients’ blood and then developed into neurons. This project will focus on a specific group of proteins, JNKs proteins, which play an important role in regulating neuronal function.

The therapeutic action of D-JNK11, a specific inhibitor of JNK was presented in a model of Alzheimer’s disease. It is important to stress that the molecule has already passed phase II clinical trials for the treatment of neurodegenerative diseases and phase III for care of hearing loss. To summarize the objective of this proposal is to investigate the involvement of JNK protein in AS and investigator the potential therapeutic effect of D-JNK11.

2. Dr. Ben Philpot
Quantifying EEG abnormalities and identify biomarkers in AS

Epilepsy is very common in patients with AS (90%) and the underlying neural alterations can be measured non-invasively through the EEG. The EEG of AS patients has a distinctive profile that allows a qualitative assessment, but not a quantitative measure that can be evaluated strictly by analytical methods.

There is the unmet need to quantify the EEG profiles in individuals with AS to deepen the understanding of the basic circuits involved in these abnormalities through studies in the mouse AS model. The hypothesis is that a study that parallels EEG in humans and mice with AS, using new clinical biomarkers will make quantitative methods possible, and at the same time be useful for understanding for the mechanisms. In collaboration with two clinics dedicated to AS in the United States, their goal is to improve clinic trials by identifying objective and impartial biomarkers.

Glossary
Pluripotent stem cell are master cells. They can potentially produce any cell or tissue the body needs to repair itself. This “master” property is called pluripotency.
White matter is one kind of brain tissue. There are 2 kinds of tissue in the central nervous system (brain and spinal cord) i.e. grey matter and white matter. Grey matter has a pinkish-grey color in the living brain, contains the cell bodies, dendrites and axon terminals of neurons, so it is where all synapses are. White matter connects different parts of grey matter to each other.
Hyper-excitability is an excessive response to a stumuli, in particular those nerve stimuli.
Proteasomes and the ubiquitination process is the main way that proteins are broken down in human cells. A breakdown in this process results in the accumulation of proteins and can be the basis of many diseases.
AZUL domain is a specific location on UB3EA gene.
PML is promyelocytic leukemia protein. This protein is involved in many cellular processes including cell cycle progression, DNA damage repair and others.
In-vivo experiments are experiments done within the living body of an animal or human.
Electroporation is the process of getting DNA or chromosomes into bacteria or other cells using a pulse of electricity to open the pores in the cell membranes/surface briefly.
Neuronal migration is the method by which neurons travel from they are created to their final position in the brain as the brain develops.
Neocortex is a part of the cerebral cortex concerned with sight and hearing in mammals, regarded as the most recently evolved part of the cortex.
Synapses is a structure that allows a nerve cell to pass an electrical or chemical signal to another nerve.

December 29, 2015

Angelman Syndrome Alliance call to research January 2016.

MORE THAN 300 000 EUROS FOR RESEARCH ON ANGELMAN SYNDROME

Call for Applications for Research Grants on  Angelman Syndrome

Call will be open between the 25th November 2015 and the 24th of January 2016

The Angelman Syndrome Alliance (ASA) supports research on Angelman syndrome with up to 300 000 Euros to be awarded at the next International Angelman Syndrome Scientific Conference. The Angelman Syndrome Alliance is (currently) an initiative of the following parent organisations: AVO (Austria), ASB (Belgium), ASFA (France), Angelman e.V. (Germany), AIA (Ireland), Israeli Angelman Syndrome Foundation (Israel), ORSA (Italy), PWAV (Netherlands), Angel (Portugal), ASSERT (United Kingdom), and the Nina Foundation (consortium organization on AS science). Their mutual aim is to join forces and funds to increase the financial support of dedicated research projects and thereby intensify research on Angelman Syndrome.

Angelman Syndrome is a genetic disorder resulting from a loss of function of the UBE3A gene which encodes the ubiquitin E3 ligase E6AP. Angelman Syndrome is characterised by severe intellectual disability, lack of speech, motor coordination deficits, sleep disturbance, and usually a happy demeanour.

Researchers of any country are invited to apply for research grants covering preclinical (basic) research as well as translational research such as mechanism-based controlled trials. Applications for behavioural therapies are not eligible at this point.
Proposals addressing new ideas/concepts about the pathogenesis of Angelman Syndrome are particularly invited.
The duration of the grant is flexible, but the maximum is 4 years. There will be an evaluation by the SAB of the project after completion of 50% of the total time and at the end. Every 6 month interim reports are to be sent to Faustine Bourgoin in simple language (lay version) with supporting pictures to inform the parent organisations in more general terms about the project progress.

The application procedure is two-step:
1. First, a short application has to be submitted on the ASA website : CLICK HERE (Deadline 24th of January 2016);
2. A maximum of 10 applicants will be selected for step 2 will be informed by end of February 2016. Selected applicants will then be invited to submit a detailed application by 3rd of May 2016.
3. 1 – 2 applicants will be awarded a grant at the bi-annual meeting of the Angelman Syndrome Alliance on the 30th of September 2016 in Portugal.
4. Via uploading their documents, the applicants accept the terms and conditions of the ASA contract (available on the website). The awardees will be expected to give the ASA-AWARD LECTURE at the beginning and the end of the project.

The short application (step 1) should include the following:
1. Cover letter including the title of the proposal and the name of the principal investigator
2. Two-page summary of the proposed research including paragraphs on the hypothesis, background, research plan, principal methodology, and the significance of the proposed research. An overview of the budget must also be included (indirect costs cannot be applied for).
3. Curriculum vitae of the applicant

All applications should be written in English language using Times New Roman, 10 point, 1.5 spaced.
Proposals involving laboratory animal testing must be approved by the respective institutional review boards before funding can be released.
All proposals, including cover letters, should be uploaded at the ASA website: angelmanalliance.org

à Questions about the grant application procedure and about the contract should be directed to the NINA Foundation at: martijn.van.steensel@ninafoundation.eu
à Questions about research on Angelman Syndrome should be directed to: harald.sitte@meduniwien.ac.at
à All general questions should be directed to: faustine.bourgoin@angelman-afsa.org

Betty Willemsen and Martijn van Steensel are part of the scientific AS organization named ‘Nina Foundation’ (NF) which is organising the call on behalf of the Angelman Syndrome Alliance (ASA). Harald Sitte (MedUni Vienna) is the chairman of the ASA Scientific Advisory Board (SAB); further members are Lidia Larizza (Univ. Milan) and Martin Scheffner (Univ. Konstanz). Faustine Bourgoin is part of AFSA, the French AS parent organization, and supports ASA and SAB with communication and tender process matters.

December 29, 2015

Glossery of scientific terms – new for 2016

Having recently sat in on a number of scientific presentations we thought it might be useful to include a new post on explaining terms. This is something we will update on a regular basis and thank Annette Kent for preparing them. Annette has come on board to support us in the area of science and research.

A target is an entity e.g. protein, gene, structure in the body that can be altered negatively or positively for the treatment of a condition/illness. The change in the target may correct the error or alleviate the problem associated with the condition/illness. Novel targets are those are being researched and have the potential to be used in future treatments.

Rescue drugs are those that used when a condition occasionally gets worse i.e. becomes acute. These medicines are not taken routinely or every day.

Orphan drugs are drugs that deemed suitable for treating very rare diseases also called orphan diseases. Orphan diseases are those that affect a small percentage of the population. These rare disease are usually genetic and present for the person’s entire life.

Proteins are very important compounds in the body and are responsible for doing a lot of the work in the cells of the body including regulating the cells activities, providing structure and functions within cells and organs of the body.

 

 

 

June 23, 2013

RTE radio 1 documentary An Angelman Year.

June 20th RTE Radio 1  – An Angelman Year.

 

May 24, 2013

Parent Conference Dublin July 20th Video sessions.

 

Angelman Syndrome Ireland Parent workshop  July 2013 video sessions  are available to view via Youtube on our home page.

Speakers sesssions:

  • Heather Lynch Director of Camphill Communities in Ireland.
  • Prof Ben Philpot, from the University of North Carolina US
  • Heather J E Crawford MBE, AHP Manager for Children’s Services/ Lead Professional for Speech & Language Therapy South Eastern Trust. NI

 

 

 

 

 

 

 

 

 

« Previous Page