Tesi sul tema "Therapy of digestive disorders"

Oxygen therapy remains a cornerstone of medical practice and is generally regarded as being safe. However, there is a lack of clinical evidence to support the routine use of oxygen therapy, and in certain conditions, injudicious oxygen may cause harm. In this thesis, I will present two audits and three randomised controlled trials of oxygen therapy. Methods  A prospective audit of the prescription and use of oxygen therapy before and after the introduction of an oxygen prescription section on a drug chart  A retrospective audit of ambulance oxygen administration, in patients with acute exacerbations of chronic obstructive pulmonary disease (AECOPD)  Two randomised controlled trials of high flow versus titrated oxygen in 150 patients with community acquired pneumonia and 106 patients with acute severe asthma  A randomised controlled trial of 24 subjects with obesity hypoventilation syndrome (OHS) comparing 100% oxygen with air Results  Oxygen prescription is suboptimal in hospital inpatients. Whilst an oxygen prescription section improved prescription, this intervention did not improve clinical practice  Over 70% of patients presenting with AECOPD received high flow oxygen prior to presentation to the emergency department. The risk of adverse outcomes increased progressively with increased PaO2  High concentration oxygen leads to a rise in PaCO2 compared to titrated oxygen, when administered to patients presenting with asthma or pneumonia  Breathing 100% oxygen leads to a rise in PaCO2 in patients with OHS Conclusion This series of studies has shown that further measures are warranted to ensure the safe practice of oxygen therapy in the pre-hospital and hospital setting. In addition, the findings suggest that the potential for high concentration oxygen therapy to increase PaCO2 is not limited to COPD but may occur in other respiratory conditions in which abnormal gas exchange or respiratory drive are present.

Introduction Epithelial homeostasis is guaranted by somatic stem cells wich, through different patways, such as p63 and Notch signaling, self-renew, differentiate and control tissue function and integrity. In vivo behavior of epithelial stem cells (ESCs) from different epithelia reflects differences in physiological role of the tissue. Limbal stem cells deficiency (LSCD) is characterized by conjunctival epithelial ingrowth, neovascularization, recurrent corneal erosion and persistent ulcers, as well as corneal scarring and ultimately leads to visual impairment and blindness. When the cornea is entirely covered with a fibro-vascular tissue, the chances of success of a traditional penetrating keratoplasty are virtually absent. Transplantation of autologous limbal epithelial stem cells (LESCs) or oral mucosa epithelial stem cells (OMESCs) cultured on fibrin glue has shown to be successful for unilateral or bilateral LSCD treatment, respectively. Despite that, fibrin presented several limitations including inability to repair or replace damaged corneal stroma. Aim The purpose of this work is to better understand the potentiality of somatic epithelial stem cells in order to identify possible approaches to exploit this potentiality. In the light of this, my research group has been able to identify and set up a cell therapy approach for a unique homozygous-heterozygous mosaicism of EEC Syndrome, demonstrating that epithelial stem cells have an intrinsic potential for regenerative medicine that can be exploited with a deeper characterization. We, also, aimed to test human keratoplasty lenticules (HKLs): particularly attractive, full-thickness scaffolds for corneal epithelial and stromal reconstruction that provides an interesting organotypic culture system for evaluation of growth, proliferation, and differentiation processes of epithelial stem cells (ESCs). Results During the three years of PhD I had the opportunity to collect and review cellular biology data of four diverse types of primary epithelial cells, derived from four different epithelia (skin, oral mucosa, limbus/cornea and conjunctiva) and, to better investigate the exhaustion of clonogenic potential and the self-renewal of epithelial stem cells we took in consideration also p63-defective oral mucosa primary cell lines obtained from three patients affected by EEC syndrome, already known to cause an acceleration in epithelial aging. Demonstrating that epithelial stem cells have an intrinsic potential for regenerative medicine that can be exploited with a deeper characterization, we have also been able to apply a cell therapy protocol to a patient affected by EEC syndrome in a rare form of mosaicism. We isolated the cell population with a mild phenotype from this patient, enriching these primary cells in vitro and producing well-organized and stratified epithelial sheets. The novelty and the importance of this case was related to the possibility to start a customized cell therapy approach for this unique case of EEC syndrome, based solely on epithelial stem cell manipulation. Limbal stem cells (H-LESCs) expanded onto HKLs gave rise to a keratinized stratified squamous epithelium morphologically similar to that of normal corneas. To set up the cohort of animal patients, we proceded with the characterization of cell lines obtained from biopsies of the cornea (C-CESCs) and oral mucosa (C-OMESCs) of canine origin. Primary lines were serially propagated until exhaustion in order to get life span data to compare their behavior with human limbal stem cells. For each passage, we also performed colony forming efficiency assays (CFE) to estimate the proportion of clonogenic cells present in the culture. Results showed a trend of canine cell lines comparable to human limbal cells trend, with a similar decrease of clonogenic cells number, a similar percentage of aborted colonies during serial cultivation and a similar replicative senescence. Then, primary human and canine epithelial stem cells were seeded onto HKLs. The resulting epithelia was well organized and stratified into four to five cell layers with basal cuboidal cells differentiating upward to winged cells. The layer of basal cuboidal cells was firmly attached to the underlying ECM and to the basement membrane through integrinβ4. Maintenance of stemness potential and differentiation pathways were assessed checking the expression of the stem cells marker p63 and the terminally differentiated cells marker Involucrine. Importantly, expression of the different markers resembled that observed in normal epithelia, thus suggesting that HKLs are able to support the growth and maintain the differentiation pathways of epithelial stem cells. Discussion and conclusions Our findings demonstrate that primary epithelial cells have unique characteristics with an inimitable potential that makes them a malleable tool, able to adapt to different necessities. And so on, although these data are intriguing, further investigation could provide more and more useful data for their clinical application in regenerative medicine. As a proof-of-principle that epithelial stem cells have an intrinsic potential for regenerative medicine that can be exploited with a deeper characterization, my group have been able to apply, in vitro, a cell therapy protocol to a patient affected by a rare mosaic form of EEC syndrome. HKLs appear to be particularly attractive, animal-free (feeder-free) and full-thickness scaffolds for corneal reconstruction. We have already started with the the recruitment of canine patients to assess the transplantability and functionality of these organotypic structures and, through the collaboration with different veterinary departments, we are creating a small cohort on which to start with first transplantation trials.
Introduzione L'omeostasi epiteliale è garantita da cellule staminali somatiche che, attraverso diversi pathways, come quello di p63 o di Notch, si auto-rinnovano, differenziano e controllano la funzione e l'integrità del tessuto. Il comportamento in vivo delle cellule staminali epiteliali (ESC) di diversi epiteli riflette le differenze nel ruolo fisiologico che queste cellule hanno nel tessuto di origine. Il deficit di cellule staminali limbali (LSCD) è caratterizzato da crescita epiteliale congiuntivale, neovascolarizzazione, erosione corneale ricorrente e ulcere persistenti, nonché cicatrici corneali, e porta infine a deficit visivo e cecità. Quando la cornea è interamente coperta da un tessuto fibro-vascolare, le probabilità di successo di una cheratoplastica penetrante tradizionale sono praticamente assenti. Il trapianto di cellule staminali epiteliali autologhe del limbus (LESCs) o di cellule staminali epiteliali della mucosa orale (OMESCs) coltivate su scaffold di fibrina ha dimostrato di avere successo per il trattamento unilaterale o bilaterale di LSCD, rispettivamente. Nonostante ciò, la fibrina presentava numerose limitazioni tra cui l'incapacità di riparare o sostituire lo stroma corneale danneggiato. Scopo Lo scopo di questo lavoro è quello di comprendere meglio la potenzialità delle cellule staminali epiteliali somatiche al fine di identificare possibili approcci per sfruttare questa potenzialità. Alla luce di ciò, il mio gruppo di ricerca è stato in grado di identificare e impostare un approccio di terapia cellulare per un caso unico di mosaicismo omozigote-eterozigote della Sindrome EEC, dimostrando che le cellule staminali epiteliali hanno un potenziale intrinseco per la medicina rigenerativa che può essere sfruttata con una caratterizzazione più profonda. Un secondo scopo della tesi è stato quello di testare gli Human Keratoplasty Lenticules (HKL): scaffolds particolarmente attraenti per la ricostruzione a tutto spessore della superficie anteriore dell’occhio che fornisce, anche, un interessante sistema di coltura organotipica per la valutazione dei processi di crescita, proliferazione e differenziazione delle cellule staminali epiteliali (ESCs ). Risultati Durante i tre anni di dottorato ho avuto l'opportunità di raccogliere e rivedere i dati di biologia cellulare di quattro diversi tipi di cellule epiteliali primarie, derivate da quattro diversi epiteli (pelle, mucosa orale, limbus/cornea e congiuntiva) e, per studiare meglio l'esaurimento del potenziale clonogenico e dell'auto-rinnovamento delle cellule staminali epiteliali abbiamo preso in considerazione anche delle linee cellulari primarie di mucosa orale con difetti genetici del gene p63 già noti per causare un'accelerazione nell'invecchiamento epiteliale, ottenute da tre pazienti affetti da sindrome EEC. Dimostrando che le cellule staminali epiteliali hanno un potenziale intrinseco per la medicina rigenerativa che può essere sfruttato con una caratterizzazione più profonda, siamo stati anche in grado di applicare un protocollo di terapia cellulare a un paziente affetto da sindrome EEC in una rara forma di mosaicismo. Abbiamo isolato la popolazione cellulare con un fenotipo lieve da questo paziente, arricchendo queste cellule primarie in vitro e producendo foglietti epiteliali ben organizzati e stratificati. La novità e l'importanza di questo caso è legata alla possibilità di iniziare un approccio di terapia cellulare personalizzata per questo caso unico di sindrome EEC, basata esclusivamente sulla manipolazione delle cellule staminali epiteliali. Le cellule staminali limbari (H-LESC) coltivate su HKL hanno dato origine a un epitelio squamoso stratificato cheratinizzato morfologicamente simile a quello delle cornee normali. Per costituire la coorte di pazienti animali, abbiamo proceduto alla caratterizzazione di linee cellulari ottenute da biopsie della cornea (C-CESCs) e mucosa orale (C-OMESCs) di origine canina. Le linee primarie sono state propagate in serie fino ad esaurimento al fine di ottenere dati di life span per confrontare il loro comportamento con le cellule staminali limbari umane. Per ogni passaggio, abbiamo anche eseguito analisi di Colony Forming Efficiency (CFE) per stimare la proporzione di cellule clonogeniche presenti nella coltura. I risultati hanno mostrato una tendenza delle linee cellulari canine comparabile all'andamento delle cellule limbari umane, con una diminuzione simile del numero di cellule clonogeniche, una percentuale simile di colonie abortive durante i passaggi in coltura e una simile senescenza replicativa. Quindi, cellule staminali epiteliali primarie umane e canine sono state seminate su HKL. L'epitelio risultante era ben organizzato e stratificato in quattro o cinque strati cellulari con cellule cuboidali basali che differenziano le cellule verso l'alto e quelle alate. Lo strato di cellule cuboidali basali era saldamente attaccato alla matrice extracellulare sottostante e alla membrana basale attraverso l'integrina-4. Sono stati valutati il mantenimento del pool di cellule staminali e i corretti processi di differenziamento controllando l'espressione del marcatore di cellule staminali, p63, e del marcatore di cellule terminalmente differenziate, Involucrina. È importante sottolineare che l'espressione dei diversi marcatori assomiglia a quella osservata negli epiteli normali, suggerendo quindi che gli HKL sono in grado di supportare la crescita e il mantenimento di tessuti epiteliali ricostruiti. Discussione e Conclusioni I nostri risultati dimostrano che le cellule epiteliali primarie hanno caratteristiche uniche con un potenziale inimitabile che le rende uno strumento malleabile, in grado di adattarsi alle diverse necessità. Sebbene questi dati siano intriganti, ulteriori indagini potrebbero fornire dati sempre più utili per la loro applicazione clinica nella medicina rigenerativa. Come prova di principio che le cellule staminali epiteliali hanno un potenziale intrinseco per la medicina rigenerativa che può essere sfruttato con una caratterizzazione più profonda, il mio gruppo è stato in grado di applicare, in vitro, un protocollo di terapia cellulare a un paziente affetto da un raro mosaicismo di sindrome EEC. Le HKL sembrano essere scaffold particolarmente attraenti (animal-free) per la ricostruzione a tutto spessore della cornea. Abbiamo già iniziato con il reclutamento di pazienti canini per valutare la trapiantabilità e la funzionalità di queste strutture organotipiche e, attraverso la collaborazione con diversi dipartimenti veterinari, stiamo creando una piccola coorte sulla quale iniziare con le prime prove di trapianto.

Thesis (D.C.P. / M. Sc.)--Faculty of Science, University of Sydney, 2006.
Submitted in fulfilment of the requirements for the degree of Doctor of Clinical Psychology/Master of Science to the Faculty of Science. Title from title screen (viewed 2 August 2007). Bibliography: leaves 95-107.

The present study examined whether or not the temporal pattern of symptom change defined as sudden gains is applicable to and has significant ramifications for understanding recovery from eating disorders. Sudden gains were defined as stable and clinically significant changes that take place between two sessions of treatment. Data for the current study were drawn from an efficacy study of CBT for eating disorders which included session-by-session measures of eating disorder symptomatology. Predictors of sudden gains were measured by an observer coded scale that included ratings of therapist use interventions, client change in behaviors and beliefs, client engagement, and homework completion. Three research questions were addressed: First, is the phenomenon of sudden gains present in CBT for eating disorders? Second, do sudden gains in CBT for eating disorders follow the three-stage model proposed for sudden gain recovery in other disorders (i.e., cognitive changes during critical sessions => sudden gains => upward spiral that includes further cognitive changes and greater long-term symptom improvement (Tang & DeRubeis, 1999b)? Third, what are the predictors of sudden gains in CBT for eating disorders that distinguish the critical session that takes place right before the sudden gain? Findings suggest that many eating disordered clients (62%) experienced at least one sudden gain during the course of CBT treatment. Three distinct types of sudden gains were identified: total symptom sudden gains, eating-related sudden gains, and body-related sudden gains. The average magnitude of these sudden gains was large representing on average 35% of total symptom improvement. Clients who experienced total symptom and body-related sudden gains demonstrated fewer eating disordered symptoms than the other clients at posttreatment. During the session preceding the sudden gain, therapists had increased levels of cognitive interventions and empathy, and clients experienced more cognitive changes and increased motivation.

Thesis (B.Sc)--University of Hong Kong, 1997.
"A dissertation submitted in partial fulfilment of the requirements for the Bachelor of Science (Speech and Hearing Sciences), The University of Hong Kong, April 30, 1997." Also available in print.

Computerised cognitive behavioural therapy (CCBT) is a clinically effective method of delivering CBT which may help address the under – treatment of common mental health disorders (CMHDs) in the population. However, concerns regarding acceptability, attrition rates and the therapeutic alliance are obstacles to widespread population dissemination. This thesis aimed to address these implementation issues by applying concepts from human – computer interaction (HCI) and attachment theory to the field of CCBT. Chapter 1 presents a meta – analysis investigating the effectiveness of CCBT for CMHDs and moderators of this effect. Chapter 2 presents a systematic review and analysis conducted to examine predictors of CCBT engagement. A process – based model of engagement with CCBT developed from the findings of this review is also presented. Adult attachment is known to influence engagement and alliance in face to face therapies, but research has not explored whether these relationships are mirrored in CCBT. Four empirical studies intended to address this question. Study 1 used a student population based survey to explore the acceptability of CCBT in a student population and the associations with adult attachment. Results demonstrated adult attachment was not associated with acceptability of CCBT. Study 2a utilised an open trial of a supported CCBT program to investigate whether adult attachment would predict engagement and alliance in vivo. Results showed attachment did not predict these outcomes. Study 2b utilised an open trial with a non – supported online CCBT program. Results indicated attachment security was positively associated with program engagement and alliance. It is proposed a combination of attachment system activation and perceiving computers as social actors account for these findings. Study 3 used a randomised, experimental paradigm to test the benefits of security priming in CCBT. Security priming produced higher levels of program engagement and better working alliance compared to neutral primes. Furthermore these effects were not moderated by dispositional attachment styles. These results demonstrate something so uniquely human, dispositional attachment orientations, founded on the intimate bonds we form in infancy and in adulthood , extend their influence into the experience of unguided CCBT, a solely human – computer interaction. Unguided – CCBT, a highly cost effective intervention with the potential for considerable public health impact, may benefit from incorporating security priming techniques in program designs to maximise engagement and alliance. Engagement and alliance is attainable in CCBT and paying attention to the attachment styles of program users may present a distinctive opportunity to overcome these implementation barriers.

Le développement récent de l’endoscopie interventionnelle digestive nous a conduit à prendre en charge deux types de pathologies préoccupantes. Il s’agit d’une part des fistules digestives souvent responsables d’une morbi-mortalité élevée et d’autre part des sténoses œsophagiennes après résection tumorale endoscopique étendue. Dans ces deux situations, des phénomènes inflammatoires chroniques conduisent soit à l’absence de fermeture de la fistule soit à une fibrose importante responsable de sténose de l’œsophage. La thérapie cellulaire a déjà été utilisée pour diminuer ces phénomènes inflammatoires et entrainer une cicatrisation. La thérapie tissulaire par cellules souches organisées en construction 3D représente un avantage important en permettant de cibler le site d’action par dépôt direct du feuillet cellularisé. Notre objectif était d’évaluer l’effet thérapeutique de ces nouveaux outils pour fermer les fistules digestives et pour prévenir la survenue des sténoses œsophagiennes. La première étape a consisté a évaluer l’efficacité du traitement par des cellules souches mésenchymateuses provenant de moelle osseuse humaine, marquées puis organisées en doubles feuillets, dans un modèle de fistule entéro-cutanée post-chirurgicale chez la souris nude. L’évaluation clinique et en imagerie (IRM et microscopie confocale) a montré une meilleure cicatrisation avec une augmentation de la microvascularisation et une accélération de la fermeture de la fistule chez les souris greffées. Les effets observés semblent liés à une augmentation précoce de la synthèse des facteurs de réparation (EGF et le VEGF) et des cytokines anti-inflammatoires (TGF-ß2 et IL-10). Après avoir développé un modèle inédit de fistule oeso-cutanée chez le porc grâce à la mise en place par voie endoscopique et chirurgicale de prothèses plastiques entre la lumière œsophagienne et la peau, nous avons évalué l’efficacité thérapeutique d’un gel contenant des vésicules extracellulaires issues de cellules souches isolées du tissu adipeux de porc. Ce gel injecté dans la fistule par voie endoscopique a permis la fermeture des fistules. Enfin, la troisième partie de notre travail a consisté à évaluer l’efficacité de la greffe allogénique de doubles feuillets de cellules souches mésenchymateuses pour prévenir la survenue des sténoses œsophagiennes dans un modèle porcin après dissection sous muqueuse étendue. Il existait une réduction significative du taux de sténose œsophagienne cicatricielle dans le groupe greffé avec une fibrose moins importante. En conclusion, l’effet paracrine antifibrosant des cellules souches mésenchymateuses organisées en feuillets est efficace à la fois pour fermer les fistules entéro-cutanées chez la souris et pour prévenir les sténoses œsophagiennes chez le porc. Un gel avec des vésicules extracellulaires issues des cellules souches a de la même façon un effet cicatrisant anti-inflammatoire permettant la fermeture des fistules œsophagiennes chez le porc. Ces résultats sont très encourageants et posent la question d’une évaluation future chez l’homme
Recent developments in digestive interventional endoscopy lead us to manage two types of digestive disease. First, it is digestive fistulas associated in many cases with high morbi-mortality; and second is oesophageal stenosis after extended superficial endoscopic resection. In both situations, chronic inflammatory process resulted in delayed or no fistula healing for the first case or oesophageal stenosis due to fibrosis. Cellular therapy has proved to be successful in reducing the inflammatory process and to promote tissue healing. Tissue therapy with 3D construct stem cells represents a major advantage by allowing a direct adaptation on the right place. Our objective was to evaluate the therapeutic effect of new strategy to close the digestive fistula and to prevent oesophageal stenosis. First step was to evaluate the effect of labelled human bone marrow derived mesenchymal stem cells engraftment in the form of double cellsheet in a post-surgical fistula model in nude mice. Clinical and radiological (MRI and probe based confocal microscopy) evaluation showed a better fistula healing with higher microvascularization and a faster fistula closing in grafted mice. These effects appear to be related to an increase production of factors involved in tissue repair (EGF et le VEGF) and anti-inflammatory cytokines (TGF-ß2 et IL-10). We developed an unpublished eso-cutaneous fistula in a porcine model after plastic catheters placement by surgical and endoscopic way between the oesophageal lumen and the skin. We evaluated the therapeutic effect of a hydrogel with extracellular vesicles extracted from porcine adipose derived stem cells. The hydrogel with vesicles was injected into the fistula by endoscopy. Radiological and histological evaluation 15 days after injection showed a fistula tract closure in treated group.The third part of this work was to evaluate the effect of allograft of adipose derived stem cells 3D construct to prevent the stenosis after extended endoscopic submucosal dissection in a porcine model. There was a significant reduction of number and degree of stenosis with decrease fibrosis infiltration in the grafted group.In summary, thanks to their paracrine and antifibrotic effect, the mesenchymal stem cells organised as 3D construct allowed fistula closure in an entero-cutaneous model in mice and prevention of stenosis after extended oesophageal submucosal dissection in a porcine model. Moreover, endoscopic hydrogel and extracellular vesicles injection allowed oesophageal fistula healing in a porcine model. These promising results pose the challenge of future clinical studies

Fear is an innate emotion and an adaptive response to provide protection from potential harm. When fear is excessive and out of proportion in relation to the confronted situation, it can lead to the development of an anxiety disorder. Many individuals feel anxious at some point, but not all experience clinical anxiety or meet the diagnostic criteria of an anxiety disorder. Still, anxiety disorders are the most prevalent form of psychiatric disorder in the general population. More often than not people suffering from one anxiety disorder also present other psychiatric conditions. As of today, cognitive and behavioural treatments have been tested and found to positively affect anxiety disorders, making them the treatment of choice. Nevertheless, many patients do not seek or receive adequate treatment. One common critique of the research trials from which the recommendations for treatments stem is the use of a single protocol targeting only one diagnosis. This is because many people suffer from comorbidities. Another problem connected to the recommendation that cognitive behavioural therapy (CBT) should be the treatment of choice for anxiety disorders is the lack of therapists with adequate training. One possible way of dealing both with the shortcoming of therapists and making CBT more accessible is the use of the Internet. Internet-based CBT (ICBT) has been tested in numerous trials during the last 15 years, showing positive outcomes for a large variety of disorders. Many ICBT trials also make use of a single protocol. Another way of dealing with comorbidities might be to tailor the treatment to let characteristics and preferences of the patient guide the design of the protocol. Little is known about possible effects of tailoring the ICBT, the effects of therapeutic relationships in ICBT, and the effectiveness and cost-effectiveness of these treatments. This thesis is based on three studies on two separate randomized controlled trials (RCTs) using the same set of modules accessible for the tailored protocol. Study I was an RCT investigating treatment effects up to two-year after completion, showing favourable outcomes of the treatment in a self-recruited sample at all measure points. Study II was a secondary analysis exploring possible relations between working alliance and treatment outcome for participants in the treatment group recruited for Study I indicating that working alliance predict outcome in this tailored treatment. The second RCT was an effectiveness trial (Study III) analysing treatment effects and cost-effectiveness of the treatment up to one year post treatment in a primary-care population. This study showed positive treatment effects both regarding symptom reduction and cost-effectiveness, and that effects were sustained at one year post treatment. Conclusions drawn from these studies are that individually tailored ICBT seems to be a feasible approach for patients with anxiety disorders regardless of comorbidities, and a responsible choice in terms of societal costs.
Rädsla är en medfödd känsla och en adaptiv respons för att skydda organismen från potentiell skada. När rädslan blir överdriven och oproportionerlig i relation till den konfronterade situationen, kan det leda till utvecklandet av ångestsyndrom. Många personer upplever någon gång ångest, men inte alla upplever klinisk ångest eller uppfyller de diagnostiska kriterierna för något ångestsyndrom. Trots detta är ångest det vanligaste psykiatriska tillståndet i befolkningen i stort och oftast uppfyller personer som lider av ett ångestsyndrom även andra  psykiatriska tillstånd. Till dags dato har både kognitiva och beteendeinriktade behandlingar testats och visat sig verksamma vid ångestproblem, vilket gjort dem till de behandlingar som rekommenderas för dessa tillstånd. Trots god effekt av behandling söker många patienter ändå inte hjälp, alternativt erhåller inte adekvat behandling. En vanlig kritik mot den forskning från vilka behandlingsrekommendationerna för ångestsyndrom stammar är att många använt en manual eller ett protokoll som riktar sig mot bara en diagnos. Detta på grund av den stora komorbiditeten. Ett annat problem kopplat till rekommendationerna att kognitiv beteendeterapi (KBT) ska vara förstahandsval vid behandling av ångest är bristen på behandlare med adekvat utbildning. Ett möjligt sätt att göra KBT mer tillgängligt är att använda Internet. Internet- förmedlad KBT (IKBT) har prövats i ett stort antal studier de senaste 15 åren dessa har visat positiva resultat vid ett stort antal psykiatriska tillstånd. Flertalet av dessa studier har dock använt ett enda behandlingsprotokoll. En annan möjlighet att hantera komorbiditet kan vara att skräddarsy behandlingen för att låta patientens egenskaper och preferenser vara med och styra utformningen av behandlingsprotokollet. Möjliga effekter av att skräddarsy IKBT är relativt lite undersökt, likaså effekterna av terapeutiska relationer i IKBT samt klinisk effektivitet och kostnadseffektiviteten för dessa behandlingar. Denna avhandling bygger på tre studier från två randomiserade kontrollerade studier med samma uppsättning av moduler tillgängliga för att skräddarsy behandlingsprotokollen. I Studie I undersöktes behandlingseffekter upp till två år efter avslutad behandling i en självrekryterad grupp patienter. Studie II var en sekundäranalys av behandlingsgruppen från Studie I där eventuella samband mellan arbetsallians och behandlingsresultat undersöktes. Den andra randomiserade kontrollerade studien var en prövning av huruvida denna behandling var effektiv för en klinisk population (Studie III) rekryterad via primärvården. Förutom behandlingseffekter undersöktes även kostnadseffektiviteten upp till ett år efter behandlingsavslut. De slutsatser som dras utifrån dessa studier är att skräddarsydd IKBT verkar vara en framkomlig väg för patienter med ångest oavsett komorbiditet, att arbetsalliansen kan vara en faktor som påverkar utfallet, samt att det är ett ansvarsfullt val vad gäller samhälleliga kostnader.

L’instabilité d’ADN mitochondrial (ADNmt) peut être quantitative avec la déplétion de l’ADNmt ou qualitative avec des délétions de l’ADNmt. Ces anomalies sont une des causes les plus commmunes des maladies mitochondriales. Un des gènes qui contrôle la stabilité et le maintien de l’ADNmt est POLG. Ce gène code pour la polymerase gamma mitochondriale. Chez l’homme, les mutations dans le gène POLG sont liées aux maladies mitochondriales telle que; l’insuffisance hépatique, le syndrome d’Alpers, le PEO ou Progressive External Ophtalmoplegia, la neuropathie sensorielle et l’ataxie. Des mutations dans le gène POLG sont aussi associées au syndrome de Parkinson. Aujourd’hui, il n’existe aucune thérapie pour ces maladies. Compte tenu de la conservation évolutive de la fonction mitochondriale de la levure à l’homme, nous avons utilisé deux organismes modèles, Saccharomyces cerevisiae et Caenorhabditis elegans, pour identifier des molecules chimiques capables de compenser l’instabilité de l’ADNmt liée à des mutations du gène POLG dans des fibroblastes d’un patient. Nous avons trouvé trois molécules candidates potentielles: MRS2, MRS3 et MRS4, à partir d’un criblage primaire chez la levure, en utilisant une chimiothèque d’environ 2000 molécules chimiques. MRS3 est la molécule candidate la plus efficace pour la stabilization d’ADNmt chez des mutants POLG de la levure, du champignon filamenteux, du nématode et sur des fibroblastes de patients. MRS3, ou clofilium tosylate (CLO), est un agent antiarrhytmique, médicament pour soigner les troubles du rythme cardiaque. Dans cette étude, nous avons aussi montré que deux autres antiarrhythmiques appartenant à la même classe que CLO avaient un effet positive chez un mutant POLG de C. elegans. En utilisant une approche de chemogénomique chez la levure, nous avons identifié Fis1, un acteur de la fission mitochondriale qui pourrait être impliqué dans la mode d’action de CLO. Fis1 est requise pour la viabilité cellulaire en concentration légèrement toxique de CLO et nécesaire pour la stabilization de l’ADNmt par CLO. L’ensemble de ces résultats ont montré que CLO pourrait être la première molécule chimique qui stimule la réplication de l’ADNmt et qui pourrait être développée pour le traitement des maladies liées à des mutations dans le gène POLG. Ces résultats ont aussi permis de mettre en évidence une nouvelle connexion entre replication de l’ADNmt et la fission mitochondriale
The instability of mitochondrial DNA (mtDNA) in form of mtDNA depletion (quantitative instability) or large deletion (qualitative instability) is one of the most common cause of mitochondrial diseases.. One of the genes responsible for human mtDNA stability, POLG, is exploited in this study. POLG encodes the human mitochondrial polymerase gamma. In human, POLG mutations are a major cause of mitochondrial disorders including hepatic insufficiency; Alpers syndrome, progressive external ophthalmoplegia, sensory neuropathy and ataxia. They are also associated with Parkinsonism. Currently, there is no effective and disease-specific therapy for these diseases. Based on the conservation of mitochondrial function from yeast to human, we used Saccharomyces cerevisiae and Caenorhabditis elegans as first pass filters to identify chemical compounds that suppresses mtDNA instability in cultured fibroblasts of a POLG-deficient patient. We found three potential candidates, MRS2, MRS3 and MRS4, from a chemical screening of nearly 2000 compounds in yeast. MRS3 is the most efficacious in stabilizing mtDNA in yeast, filamentous fungi, worm and patient fibroblasts. This unsuspected compound, clofilium tosylate (CLO), belongs to a class of antiarrhythmic agents for cardiovascular disease. Two other antiarrhythmic agents (FDA-approved) sharing common pharmacological properties and chemical structure with CLO also show potential benefit for POLG deficiency in C. elegans. Using a chemogenomic approach in yeast, we also discovered that a mitochondrial fission actor Fis1 is implicated in the mechanism of action of CLO. Fis1 is important for cellular viability in a slightly toxic concentration of CLO and is required for the mtDNA stabilizing potency of CLO. Our findings provide evidence of the first mtDNA-stabilizing compound that may be an effective pharmacological alternative for the treatment of POLG-related diseases and uncover a new connection between the mitochondrial fission process and mtDNA replication
Ketidakstabilan DNA mitokondria (mtDNA) dalam bentuk pengurangan kopi mtDNA di dalam sel (ketidakstabilan kuantitatif), atau pun dalam bentuk delesi pada sekuens mtDNA (ketidakstabilan kualitatif) merupakan salah satu penyebab penyakit mitokondria. Salah satu gen yang bertanggung jawab dalam menjamin kestabilan mtDNA adalah POLG. Gen POLG mengkode protein polimerase gamma pada manusia, yang mereplikasi dan mereparasi mtDNA di dalam mitokondria. Mutasi pada gen POLG dapat menyebabkan penyakit kelainan mitokondria pada manusia, seperti gagal ginjal, sindrom Alpers, Progressive External Ophtalmoplegia, neuropati sensorial, ataxia dan bisa dikaitkan dalam beberapa gejala Parkinsonisme. Saat ini, belum ada terapi obat yang dapat mengatasi penyakit – penyakit tersebut. Berdasarkan kesamaan evolutif dari ragi hingga manusia, pada studi ini kami menggunakan Saccharomyces cerevisiae dan Caenorhabditis elegans untuk mengidentifikasi molekul obat yang berpotensi mengatasi ketidakstabilan mtDNA dari fibroblas pasien manusia yang memiliki mutasi gen POLG. Kami mengidentifikasi tiga kandidat potensial, yakni MRS2, MRS3 dan MRS4 dari penapisan kurang lebih 2000 molekul obat dengan menggunakan ragi. MRS3 adalah kandidat yang paling berkhasiat dan mampu mengatasi ketidakstabilan mtDNA pada ragi, Podospora, cacing dan fibroblas manusia. MRS3 adalah alias bagi clofilium tosylate (CLO), sebuah molekul antiaritmia untuk penyakit kardiovaskuler. Pada studi ini, kami juga menguji aktifitas dua molekul antiaritmia lain yang tergabung dalam kelas yang sama dengan CLO, dan menemukan bahwa kedua molekul ini juga berpotensi mengatasi defisit POLG pada cacing C. elegans. Dengan menggunakan metode kemogenomik pada ragi, kami juga mengidentifikasi sebuah aktor prosesus pembelahan mitokondria, Fis1, yang berpotensi terlibat dalam mekanisme seluler CLO. Fis1 dibutuhkan untuk: (1) kelangsungan hidup ragi pada konsentrasi toksik CLO dan (2) efek CLO dalam menstabilkan mtDNA pada ragi. Keseluruhan studi ini membuktikan potensi CLO sebagai molekul penstabil mtDNA yang pertama, yang dapat dikembangkan sebagai salah satu alternatif terapi obat untuk penyakit – penyakit mitokondria terkait mutasi POLG. Melalui studi ini, juga diungkap adanya hubungan antara kestabilan mtDNA dan prosesus pembelahan mitokondria

Quality substance abuse treatment is needed to help fight the battle against drug addiction. This qualitative study was designed to explore some of the approaches to eye movement desensitization (EMDR) therapy that therapists trained in Parnell's adapted EMDR model use in conjunction with treatment for addictions. The purpose of this narrative inquiry was to investigate the experience of therapists who incorporate substance abuse treatment with Parnell's adapted EMDR model when treating trauma and substance use disorders. The population studied comprised licensed mental health therapists who had completed Parnell's EMDR training and implemented Parnell's modified EMDR protocol in their professional practice. The data from 9 participant interviews were coded and NVIVO data analysis software was used to identify key concepts and themes including deviations from Parnell's modified protocol, incorporating addiction treatment within the modified protocol, and the importance of the resourcing phase in the modified protocol. The study findings provided a deeper understanding of the types of addiction therapies that therapists are using in conjunction with Parnell's EMDR model. The results also showed that that participants perceived Parnell's EMDR model, combined with addiction therapeutic techniques and approaches, as beneficial in treating those with trauma and substance use disorders. By integrating addiction therapies with Parnell's EMDR protocol, EMDR certified trainers may better educate EMDR trainees about useful strategies for treating dual diagnosed clients. The strategies may shorten the client's time in treatment and provide a strong foundation for therapists as they conduct therapy for dual diagnosed people.

Introduction: Differentiating between normal language variation and abnormal language development can be difficult for clinicians working with young children who present with slow language development, so-called “late talkers”. Although the language difficulties of many late talkers resolve spontaneously, there is clearly a group of children whose problems persist, either for a long period or possibly permanently (after the age of five, often referred to as children with specific language impairment: SLI). There is a lack of research examining potential markers of language difficulties in young children which may enable the early detection of children at risk of SLI. The aim of this thesis was to determine the most suitable measure, or combination of measures, that can predict which late talkers at age 3;0 to 4;0 will be likely to have SLI at age 4;0 to 5;0. Methods Forty seven late talkers and 47 children with typical language development (TLD) aged from 3;0 to 4;0 were assessed on a number of language, IQ and marker tasks (baseline assessment). The children were recruited from 13 nurseries and one family centre in Aberdeen city. The children were reassessed one year later on a number of language, IQ and marker tasks (follow-up assessment). Results: Characteristics of the child or family examined were not associated with membership of the expressive language delay group at follow-up. Within the late talker group only, the Preschool Language Scale-3 Expressive Communication (PLS-3 EC) and Recalling Sentences scores at baseline were the best predictors of persistent expressive language delay at follow-up. Late talkers performance on the PLS-3 EC and Recalling Sentences tasks at age 3;0 to 4;0 has potential as predictors of persistent expressive language delay (children likely to have SLI) at age 4;0 to 5;0. Conclusions: A sizeable proportion of children identified as late talkers at age 3;0 to 4;0 have persistent language problems a year later at follow-up. The language measure PLS-3 EC has the potential to differentiate between late talkers who are going to have more persistent problems from those who recover. The marker task Recalling Sentences also has the potential to differentiate between late talkers who are going to have more persistent problems from those who recover. Given the simplicity of Recalling Sentences, this task has the potential to be a useful screening test in clinical practice although this needs to be evaluated in further research.

Current therapies for bone fragility disorders such as Osteogenesis Imperfecta (OI) can reduce fracture risk by improving bone quantity but not bone quality. Cell and/or gene therapy strategies hold promise for addressing the fundamental deficiencies in genetic bone disease but involve a number of technical hurdles that need to be overcome. This thesis takes a stepwise approach to address some of these challenges. Allogenic bone marrow transplantation (BMT) from healthy donors has been suggested for several decades to be able to repopulate the bone compartment with genetically healthy cells. However prior attempts have often featured poor osteoblastic engraftment. We describe the application of cell therapy to the mild-moderate severity Col1a2G610C OI mouse model. The effects of sub-lethal irradiation followed by transplantation of BMT from wild type (WT) mice into OI mice were analysed via DEXA, microCT, and mechanical testing. No differences were observed between the OI transplanted with WT cell group and the naïve WT and OI control groups in any measure. OI cells transplanted into OI mice were also included an additional control group to test for the paracrine effects of BMT, but again no significant differences were found compared to naïve OI controls. Lineage tracking using mice irradiated then transplanted with fluorescently labelled bone marrow cells revealed that most engrafted donor cells expressed the osteoclast marker tartrate-resistant acid phosphatase (TRAP). These results together indicate the inefficacy of irradiation and BMT on the osteopoietic compartment and suggest that alternative novel methods would be needed to increase engraftment for OI. In order to facilitate gene therapy approaches for OI, we aimed to engineer a system allowing the specific targeting of bone cells (osteoblasts and osteocytes) throughout the skeleton. Adeno associated viruses (AAVs) emerged as a prime vector candidate due to their small size, non-immunogenicity, and tissue specificity. A panel of 18 AAV variants expressing Cre recombinase and GFP under CAG ubiquitous promoter were first trialled via local delivery in a murine fracture model and in vitro on a human osteoblastic cell line. High performing variants, AAV8 and AAV-DJ were then used in systemic delivery experiments where vectors driving Cre expression via bone-cell specific promoters were designed and generated. The AAV8-Sp7-Cre vector was demonstrated to specifically and efficiently transduce osteoblasts and osteocytes throughout the skeleton. In a final series of experiments, delivery methods for the Cre constructs were compared and CRISPR/Cas9 gene editing constructs were designed and generated based on the Cre constructs design. Intraperitoneal (IP) delivery of the AAV8-CAG-Cre construct showed a similar transduction profile to intravenous (IV) delivery throughout the organs and bones. The one exception was skeletal muscle where IP delivery was able to transduce some skeletal muscle surrounding the tibia. In utero delivery was also trialled via IV delivery to pregnant female mice at ED17. This did not result in transduction of the pups, and IP injection of the pregnant female mice or direct injection of the pups in utero should be trialled. Finally, two AAV8 CRISPR/Cas9 constructs (a self-assembling intein system) able to drive gene editing of the Ai9 locus were produced. Preliminary studies showed a lack of gene editing in target tissues, and hence further studies were conducted to troubleshoot the constructs. HEK293 cells transduced with the virus in vitro showed staining of the N-terminal Cas9 intein, however the C-terminal Cas9 intein has yet to be validated. Further studies will be taken to resolve issues with these constructs to produce vectors able to mediate global skeletal gene editing in the Ai9 mouse. In summary, the published papers and subsequent experiments detailed in this thesis represent a stepwise approach for developing a gene therapy solution to genetic bone diseases. The creation of a bone specific Cre expressing AAV vector is expected to have remarkable utility as a tool for generating timed bone specific knockouts in floxed mouse lines. Its specificity and efficiency are particularly notable features. It is anticipated that rectification of one or more of the components of the split CRISPR/Cas9 approach will ultimately enable high-efficiency gene editing in bone, which will be a major advance for the field.

Dizziness is very common, but it is never normal. Dizziness can make performing daily activities, work, and walking difficult. Many people get dizzy when they turn their head, which can cause problems with walking and makes people more likely to fall. Most of the time dizziness is not from a life-threatening disease. Often dizziness is because of a disorder of the vestibular (or inner ear balance) system. People can get vestibular disorders from infections in the ear, problems with the immune system, medications that harm the inner ear, and rarely from diabetes or stroke because of a lack of blood flow to the inner ear. Stress, poor sleep, migraines, overdoing some activities, and feeling sad can increase symptoms. New guidelines for the treatment of vestibular disorders were published in the April 2016 issue of the Journal of Neurologic Physical Therapy. The guideline describes which exercises are best to treat the dizziness and balance problems commonly seen with an inner ear disorder.

Sixteen couples in which one of the partners met criteria for Somatoform Disorder or Undifferentiated Somatoform Disorder as determined by the SOMS and who scored less than or equal to 101 on the Dyadic Adjustment Scale participated in this study. These couples were randomly assigned to 8 sessions of emotionally focused therapy or to a 12-week wait list condition. The purpose of the study was to investigate the effectiveness of emotion focused therapy as a treatment of somatoform disorders. Results suggest that the intervention of emotion focused couples therapy (EFT) was not effective in the treatment of somatoform disorders. A significant effect (.023) was found in the increased reporting of symptoms at posttest. Implications for EFT and marriage and family therapy are discussed.
Ph. D.

Thesis (B.Sc.)--University of Hong Kong, 2003.
"A dissertation submitted in partial fulfilment of the requirements for the Bachelor of Science (Speech and Hearing Sciences), The University of Hong Kong, April 30, 2003." Includes bibliographical references (p. 28-30) Also available in print.

Research on art therapy and psychosis has typically focused on individuals who have experienced psychotic symptoms for many years. This study used a grounded theory methodology to explore how service users experience art therapy following their first diagnosis of a psychotic disorder, and the possible mechanisms through which art therapy might be helpful for such individuals. Eight participants were interviewed, with two participants being interviewed twice. A preliminary theory was created and seven categories were constructed from the data, namely unpressured atmosphere, pleasure and engagement in art-making, expression and communication, connecting with others, changing emotional experience and experience of self, supporting recovery and continuation of art, and barriers. Participants reported that through the atmosphere of art therapy, art-making, and communication, they were able to build relationships, connect with others, experience a sense of commonality, absorption, sense of freedom and discover alternative perspectives and different understandings. Whilst this study suffered from some limitations, the results build on the current research base by suggesting possible processes and mechanisms through which art therapy is helpful, and focusing on a previously under-represented population. The findings are considered alongside existing research and theoretical perspectives. Clinical implications and recommendations for future research are also highlighted.

Amniotic membrane is applied to the diseased ocular surface to stimulate wound healing and tissue repair, because it releases supportive growth factors and cytokines. These effects fade within about a week after application, necessitating repeated application. Generally, amniotic membrane is fixed with sutures to the ocular surface, but surgical intervention at the inflamed or diseased site can be detrimental. Therefore, we have developed a system for the mounting of amniotic membrane between two rings for application to a diseased ocular surface without surgical intervention (sutureless amniotic membrane transplantation). With this system, AmnioClip, amniotic membrane can be applied like a large contact lens. First prototypes were tested in an experiment on oneself for wearing comfort. The final system was tested on 7 patients in a pilot study. A possible influence of the ring system on the biological effects of amniotic membrane was analyzed by histochemistry and by analyzing the expression of vascular endothelial growth factor-A (VEGF-A), hepatocyte growth factor (HGF), fibroblast growth factor 2 (FGF 2) and pigment epithelium-derived factor (PEDF) from amniotic membranes before and after therapeutic application. The final product, AmnioClip, showed good tolerance and did not impair the biological effects of amniotic membrane. VEGF-A and PEDF mRNA was expressed in amniotic membrane after storage and mounting before transplantation, but was undetectable after a 7-day application period. Consequently, transplantation of amniotic membranes with AmnioClip provides a sutureless and hence improved therapeutic strategy for corneal surface disorders.

The purpose of this thesis was to assess the agreement between youths and therapists on homework adherence. Kappas were computed for sessions 4 through 12. Agreement ranged from poor (κ = .22) in session 4 to excellent (κ = .83) in session 6, with percent agreement ranging from 69.9% to 95.6%. All other sessions fluctuated from excellent to fair to good. Additionally, homework completion was examined as a function of youth characteristics according to both reporters. An independent sample t-test determined there were no significant differences in percent completion according to both reporters, except for parent income according to therapists. Possible explanations for discrepancies in agreement are discussed. It is suggested that in order to avoid discrepancies between youth and therapist reports, it may be necessary for therapists to clearly set rules and enlist the help of parents in order to ensure homework completion throughout therapy.

A qualitative, art-based research approach is used in this study to explore the relationship between trauma and eating disorders in art therapy treatment. The literature review illuminates the function, presentation, and treatment of eating disorders and trauma. Adolescent developmental challenges are discussed in relation to eating disorders, given that this is the period when the diagnosis manifests. The nominal research on the use of art therapy for underlying trauma with clients who are in treatment for eating disorders suggests the importance of considering the implications for the field of art therapy. Two-semi-structured, art-based interviews are the primary data gathering component in understanding the connection between eating disorders and trauma. An axial coding process is used to analyze the textual and visual data to reveal emergent themes. The results of the analysis process suggest that art, which bypasses verbal defenses, allows the client to access and externalize internal experiences such as trauma. The study also reveals the interconnected nature of eating disorders and trauma. The findings recommend future study of this relationship and the necessity of addressing significant traumatic experiences in addition to abuse and most importantly to acknowledge trauma as a primary focus of treatment.

Includes bibliographical references.
This study aimed to determine whether highly active antiretroviral therapy (HAART) improved cognitive function in HIV positive people in South Africa, and whether this effect differed according to the CNS penetration-effectiveness (CPE) of the regimen used. I also investigated potential HIV-Associated Neurocognitive Disorders (HAND) biomarkers (serum neopterin, osteopontin and neurofilament H) to determine their relationship to the severity of cognitive impairment at baseline in HAART-naïve patients, and whether initial levels of these biomarkers related to the change in cognitive function a year later.

The purpose of this study was to examine the characteristics that distinguish children with behavior disorders who drop out of psychotherapy treatment from those who remain in treatment. The sample included 379 children (268 male and 111 female) who were diagnosed with a behavior disorder at Dallas County Mental Health/Mental Retardation (MH/MR), a community mental health clinic in Dallas, Texas. The results indicated that certain characteristics increased the likelihood that a child would drop out of therapy, including reliance on aid, the presence of maternal psychopathology, and more severe externalizing and internalizing behaviors. This study also found that younger children with behavior disorders had a greater probability of dropping out of treatment. Minority status, gender, parent marital status, and referral source were not found to be associated with dropping out of treatment. Future studies should focus on specific interventions that clinicians could employ to deter premature termination from treatment.

The study assessed the evidence for the efficacy of lithium in the acute treatment of prophylaxis of unipolar depression and as an augmentation drug in treatment resistant depression. It examined the relationship between hormonal changes, lithium discontinuation and relapse in bipolar illness. Finally, it investigated the addition of inositol in the diet of bipolar patients as an alternative treatment for lithium side effects. AIMS: 1) to assess the efficacy of lithium in unipolar depression by comparing readmission rates in patients after lithium discontinuation and in patients who continued to take lithium as a prophylactic agent, and evidence for a lithium withdrawal syndrome in unipolar patients during the first three months of discontinuation. 2) to analyze quantitatively the results of lithium trials in the acute treatment and prophylaxis of unipolar depression in order to estimate the therapeutic value of lithium by measuring an overall effect size. 3) to investigate the neuroendocrine changes involving thyroxine (T4) thyroid stimulating hormone (TSH) and cortisol before and after lithium discontinuation in bipolar patients and to test whether they are related to the withdrawal syndrome observed in these patients. 4) to evaluate the effects of adding inositol to the diet of clinically stable bipolar patients on prophylactic lithium upon thyroxine (T4) and thyroid stimulating hormone (TSH) concentrations and on other peripheral side-effects of lithium, comparing the results to those found in a group of normal controls not taking lithium. 5) to assess the efficacy of lithium augmentation in treatment resistant depression by reviewing, using quantitative techniques, the controlled trials on the topic. While the results support the case for lithium augmentation in treatment-resistant depression, there remains considerable uncertainty over the duration of treatment necessary to see and sustain a response to lithium augmentation.

La mancanza di tecnologia per la manipolazione diretta su scala globale del sistema nervoso del topo adulto ha ostacolato la ricerca sui processi e disfunzioni cerebrali. Attualmente, il trasferimento genico è normalmente ottenuto mediante iniezioni virali intraparenchimali, ma queste iniezioni mirano a un'area del cervello ristretta. Qui, abbiamo dimostrato che la somministrazione endovenosa di un vettore virale adeno-associato (AAV) -PHP.B ha permeato e diffuso in tutto il parenchima neurale, trasducendo sia il sistema nervoso centrale e periferico in un modello globale. Abbiamo quindi stabilito molteplici procedure di trasduzione virale per controllare l'espressione genica o disattivare la funzione genica esclusivamente nel sistema nervoso adulto e valutato gli effetti comportamentali. Sulla base di questi risultati, abbiamo stabilito un'efficace strategia di terapia genica per contrastare l'accumulo diffuso di depositi di α-sinucleina in tutto il cervello in un modello murino di sinucleinopatia. Trasduzione di topi transgenici A53T-SCNA con AAV-PHP.B-GBA1 ha ripristinato i livelli fisiologici di Glucocerebrosidasi, ha ridotto l’accumulo di α-sinucleina e ha prodotto un significativo recupero comportamentale. Infine, abbiamo fornito prove che la penetrazione cerebrale di AAV-PHP.B non comporta evidenti disfunzioni nell'integrità o permeabilità della barriera ematoencefalica. Complessivamente, la strategia basata sul vettore virale AAV-PHP.B consente l'espressione neurale globale non invasiva, diffusa e duratura di geni terapeutici, come GBA1, fornendo un approccio inestimabile per il trattamento di malattie neurodegenerative con patologia cerebrale diffusa come le sinucleinopatie.
The lack of technology for direct global-scale targeting of the adult mouse nervous system has hindered research on brain processing and dysfunctions. Currently, gene transfer is normally achieved by intraparenchymal viral injections, but these injections target a restricted brain area. Herein, we demonstrated that intravenous delivery of adeno-associated virus (AAV)-PHP.B viral particles permeated and diffused throughout the neural parenchyma, targeting both the central and the peripheral nervous system in a global pattern. We then established multiple procedures of viral transduction to control gene expression or inactivate gene function exclusively in the adult nervous system and assessed the underlying behavioral effects. Building on these results, we established an effective gene therapy strategy to counteract the widespread accumulation of α-synuclein deposits throughout the forebrain in a mouse model of synucleinopathy. Transduction of A53T-SCNA transgenic mice with AAV-PHP.B-GBA1 restored physiological levels of the enzyme, reduced α-synuclein pathology, and produced significant behavioral recovery. Finally, we provided evidence that AAV-PHP.B brain penetration does not lead to evident dysfunctions in blood-brain barrier integrity or permeability. Altogether, the AAV-PHP.B viral platform enables non-invasive, widespread, and long-lasting global neural expression of therapeutic genes, such as GBA1, providing an invaluable approach to treat neurodegenerative diseases with diffuse brain pathology such as synucleinopathies.