Dissertations / Theses on the topic 'Skin Diseases Treatment Thailand'

Controlled drug delivery systems are becoming increasingly interesting with the contribution of nanotechnology. In the case of transdermal applications the greatest limitation is the highly impermeable outermost layer of the skin, the stratum corneum. One promising method of controlled transdermal drug delivery of the skin therapeutics is the use of nanoparticles as carriers. Encapsulation of the drug, as opposed to classical topical application of creams or emulsions, allows the drug to diffuse into hair follicles where drug release can occur in the deeper layers of the skin. The aim of this study was to develop micro and nano sized carriers as drug delivery systems to achieve treatment for skin conditions like psoriasis, aging or UV damage, caused by radiation or health problems. Two different types of bioactive agents, retinyl palmitate (RP) and Dead Sea Water (DSW), were used by encapsulating in poly(3-hydroxybutyrate-co-3-hydroxyvalerate) carriers. In some tests MgCl2 was used as a substitute for DSW when quantification was needed. Bioactive agent loaded nanospheres and nanocapsules were prepared with o/w and w/o/w methods in low micron (1.9 µ
m), mid nano (426 nm) and nano (166 nm) sizes. Loading, encapsulation efficiency and release kinetics were studied. The encapsulation efficiency and loading values are low especially for the water soluble agents, DSW and MgCl2. It was observed that the capsules loaded with hydrophilic agents released their content in the first 24 h in aqueous media. The encapsulation efficiency and loading values for RP were higher because of the insolubility of the agent in water. In the in vitro studies carried out with L929 mouse fibroblast cells, the nano sized PHBV capsules were detected in the cytoplasm of the cells. Cell viability assay (MTT) for L929 cells showed a growth trend indicating that the particles were not cytotoxic and the values were close to the controls. Hemolytic activity was examined using human erythrocytes and micro/nanoparticles of PHBV were found to be non hemolytic. In vivo testing with BALB/c mice, nanocapsule penetration revealed that a small amount of nano sized particles penetrated the mice skin, despite the highly impermeable outer skin layer. As a result, PHBV micro/nanoparticles have a significant potential for use as topical drug delivery systems in the treatment of skin diseases.

La pioglitazona (PGZ) es un agonista del peroxisoma proliferador activado del receptor gamma (PPARγ) receptor, pertenece a la clase de las tiazolidindionas y es usado para el tratamiento de la diabetes tipo 2. Muchos estudios han reportado el uso de este fármaco con diferentes funciones, tales como: antiinflamatoria, antiangiogénica, antifibrótica, antitumoral y neuroprotectora. Además, este fármaco ha sido investigado debido a su capacidad de reducir la respuesta inflamatoria por diferentes mecanismos. Con el fin de obtener formulaciones apropiadas de PGZ que mejoren su liberación, esta tesis tuvo como principal objetivo el desarrollo y la caracterización de formas de dosificación de PGZ (nanopartículas (NPs) poliméricas de PLGA-PEG y formulaciones líquidas) capaces de reducir la inflamación asociada con enfermedades dérmicas (rosácea), oculares (uveítis) y neurodegenerativas (Alzheimer). Estos sistemas fueron optimizados y caracterizados fisicoquímicamente, analizando las interacciones fármaco-vehículo por métodos espectroscópicos y térmicos. Estudios de estabilidades fueron llevados a cabo durante 3 meses. Se estudió también la liberación in vitro, la permeación ex vivo en diferentes membranas biológicas, la tolerancia y la distribución del fármaco en los diferentes tejidos. Asimismo, estudios de toxicidad se realizaron en líneas celulares de queratinocitos (HaCat), retinoblastoma (Y-79) y células endoteliales del cerebro (hCMEC/D3). Estudios de internalización, transporte y permeabilidad de las NPs-PGZ fueron llevados a cabo en hCMEC/D3. Para finalizar se evaluó también la eficacia terapéutica in vivo de las formulaciones desarrolladas para rosácea, uveítis y Alzheimer. Las NPs-PGZ mostraron un tamaño promedio de partícula menor a 200 nm, una polidispersión aproximada de 0.1, carga superficial de -13.6 mV y una eficiencia de asociación en torno al 92 %. Los estudios de interacción fármaco-vehículo confirmaron que la PGZ fue encapsulada en la matriz polimérica. Las formas farmacéuticas fueron estables 3 meses, con un perfil reológico newtoniano. Los estudios de liberación de las formulaciones mostraron perfiles cinéticos de primer orden e hiperbólico. En los ensayos de permeación en la piel, la solución de PGZ con un promotor fue capaz de penetrar en la piel con altos valores para flujo (Jss), cantidad retenida del fármaco (Qret) y concentraciones predichas en estado de equilibrio estacionario (Css). Además, en los estudios de permeación en las distintas mucosas (bucal, sublingual, nasal e intestinal), dependiendo de la forma farmacéutica y en base a los resultados de Css, ha sido posible predecir que pueden proporcionar un efecto farmacológico sistémico tanto a nivel de la mucosa nasal como de la intestinal. Por otro lado, en el ojo la PGZ a partir de las NPs mostró altas concentraciones permeadas a través de la esclera. Los ensayos de tolerancia in vivo (Draize test) indicaron que la solución de PGZ no producía daño en la piel. Además, las NPs-PGZ no indujeron ninguna irritación a nivel ocular tanto in vivo como in vitro por el (HET CAM test). También las fórmulas fueron testadas mostrando no toxicidad a las concentraciones determinadas en las diferentes líneas celulares. Las NPs fueron capaces de internalizar, atravesar y no alterar la permeabilidad en un modelo in vitro de la barrera hematoencefálica con las células hCMEC/D3. Con respecto a la eficacia terapéutica de las formulaciones: la solución de PGZ disminuyó notablemente los niveles de eritema después de la inducción de la inflamación en las espaldas de los ratones BALB/c en todos los intervalos de tiempos analizados, corroborado por estudios histológicos. En los estudios de eficacia ocular en cerdos, las NPs-PGZ fueron capaces de prevenir la inflamación ocular después de la inducción de la inflamación con araquidonato sódico. Las NPs-PGZ también fueron testadas (in vivo) en un modelo de ratón transgénico con la enfermedad del Alzheimer y los resultados mostraron que las NPs fueron capaces de reducir el déficit de memoria cuando se comparan con el vehículo. Además, mostró clara tendencia a reducir las placas de la proteína beta amiloide. En resumen, las formulaciones de PGZ desarrolladas podrían constituir en un futuro, después de ensayos clínicos, una nueva indicación terapéutica para este fármaco.

Due to the immuncompromised status of AIDS patients, secondary infections and malignancies are common. Conditions secondary to AIDS for which patients require treatment include Karposi's sarcoma (treated with methotrexate), varicella-zoster (treated with antivirals such as acyclovir) and herpes simplex (also treated with antivirals like acyclovir or idoxuridme). However the clinical efficacy of these drugs is limited by poor skin permeability. Few reports, however, have dealt with the delivery of low molecular weight hydrophilic drugs from these vesicles (El Maghraby et al, 2000). The aim of our study was to investigate in vitro permeation of methotrexate, acyclovir and idoxuridine across human epidermal membrane from elastic liposomes. The intent was to establish whether formulation of these hydrophilic drugs into elastic liposomes would enhance their skin permeation parameters. We developed and validated high-performance liquid chromatographic techniques for quantitative analysis of methotrexate, idoxuridine and acyclovir. Elastic liposomes were prepared from various phospholipids- phosphatidylcholine 78.6%; phosphatidylcholine 50%; hydrogenated phosphatidylcholine 90%; phosphatidylcholine 95% and surfactants - sodium cholate, sodium deoxycholate, Span 20, 40, 60, 80. These vesicles were characterised by transmission electron microscopy. The solubilities of methotrexate, acyclovir and idoxuridine were determined. Phospholipon G (95% phosphatidylcholine) was chosen for the preparation of the liposomes with different surfactants. Permeation of methotrexate, acyclovir and idoxuridme from these vesicles across human epidermal membrane was investigated. Flux values for methotrexate, acyclovir and idoxuridine values (J) obtained by curve-fitting of data using Easyplot were compared to those obtained by linear regression. We used Student's t-test to determine statistically significant differences in the flux values of the formulations. A computer program http://www.physics.csbsju.edu/stats/ttest- bulk-form.html was used for this purpose. Our results indicate that there are no statistically significant differences between flux values from elastic liposomes and saturated aqueous solutions.
Thesis (Ph.D. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2004.

The use of medicinal plants for treatment of skin infections and improvement of skin tone was assessed. The study covered Amathole and OR Tambo municipalities of the Eastern Cape Province of South Africa. The study sought to identify and document some medicinal plants which are used by the indigenous people on the skin, evaluate some biological properties which attribute to their use for therapeutic use. This report provides ethnobotanical data on some plant which were identified. Six medicinal plants (Kniphofia drepanophylla, Gnidia capitata, Hypoxis hemerocallidea, Syzgium cordatum, Macaranga capensis and Protorhus longifolia) which were implicated for treatment of skin diseases were screened for their antibacterial and antioxidant properties. Guided questionnaires were used to interview and gather ethnobotanical information from the traditional healers. Members of the communities which were indicated to be knowledgeable on use of medicinal plants in the areas were visited and interviewed. The results revealed that a total of 45 plant species distributed in 41 Genera belonging to 28 Families are used for treatment of skin infections and improvement of skin texture. The plant medicines were reported to be used as remedies against common skin problem such as wounds, pimples, acnes and itches. Some were reported to have anti-inflammatory effect on the skin while other were reported to oil dry skin and prevent skin dryness and dry eczematous conditions. Others like Cassipourea flanaganii, and Spirostachys africana were commonly used to enlighten skin and protect skin from ultra-violet radiation. The most common mode of preparation of these medicinal plants was pastes or decoctions. Sometimes crushed plant materials were put in bathing water or boiled and steamed on affected parts. It was reported that mixing pastes with oil, animal fat and milk improve medicinal properties. Water, methanol and acetone extracts of K. drepanophylla, G. capitata, H. hemerocallidea, S. cordatum, M. capensis and P. longifolia were screened against five strains of Gram negative bacteria. The results revealed that water extracts from five plant species inhibited two or more strains of bacteria with the most common minimum inhibitory concentrations (MICs) ranging between 5,0 to 10,0 mg/ml with the exception of aqueous extracts of K. drepanophylla which failed to inhibit all strains of bacteria. The screening of methanol extract of these plants for antioxidant and free radical activity revealed a significant activity with 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) free radical (DPPH). The extracts of P. longifolia, G. capitata, M. capensis, S. cordutum and H. hemerocallidea showed significantly higher free radical scavenging activity with IC50 of ranging from about 11.0 to 41.2 μg/ ml while the IC50 value for K. drepanophylla could not be determined at 100 μg/ ml. Generally the study showed that medicinal plants still play a very important role in the health care delivery system, especially in the O R Tambo and Amathole municipalities of the Eastern Cape. Botanical medicine remains pivotal in the treatment of skin ailments and improvement of the skin tone.

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas
A psoríase é uma doença crónica inflamatória da pele muito comum que atinge aproximadamente 2% da população mundial. Esta possui uma etiologia multifactorial, com envolvimento genético, imunológico e factores ambientais que despoletam a patologia. Carateriza-se principalmente pela hiperproliferação dos queratinócitos e um infiltrado de leucócitos na epiderme, além da expansão vasculares e alteração na produção de citoquinas na derme. As áreas afectas causam incómodos físicos e problemas sociais, afectando a qualidade de vida pois, apesar da doença não ser contagiosa, os portadores sentem-se constrangidos devido à aparência provocada pelas lesões. O diagnóstico baseia-se no histórico e nos sintomas clínicos do paciente, contudo, pode-se utilizar a biopsia para a confirmação deste e prevenção e tratamento apenas retardar as redicivas. Este tratamento passa, essencialmente, por utiliza agentes terapêuticos tópicos ou sistémicos, biológicos e foto/quimioterapia, não visando a cura e apenas mantendo a doença em remissão ou com melhoras do bem-estar e da qualidade de vida do paciente. O seguinte trabalho tem como objectivo principal discorrer sobre o estado da arte actual da psoríase, assim como os principais elementos da imuno-patologia e tratamento da mesma. Foi realizada uma revisão bibliográfica da literatura existente, utilizando artigos com o maior número de palavras-chave em comum e similaridade na temática. Although litlle is know about it, psoriasis is a common inflammatory chronic skin disease that reaches around 2% of the worldwide population. Is has multipathogenic aspects with genetic involvement, immunological and environment factors. The main characterisitics of psiorisis are hiperproliferation of the keratinocytes and lymphocytes infiltrated in the epidermis, and the vascular expansion and alteration in the production of cytokines in the derme. The affected areas can also cause physical discomfort and social ability problems, affecting the quality of life. Even though the disease is not contagious, patients tend to feel awkward due to the skin appearance, as a result of the injuries. The diagnosis is based on the description and on the clinic sumptoms of the patient; however the biopsy for confirmation can be also used. The prevention consists only on delaying the returns of it. The treatment that uses topical or systemic therapeutical agents, biological and photo/chemotherapy, does not aim the cure, only keeping the disease in remission improving the welfare of the patient. The choice of the treatment depends on the presentation and severity of the disease, the cost/benefit and the desire for quality of life of the patient. The following work has as main objective discuss the current state of the art of psoriasis, as well as key elements of immune-pathology and treatment of the same. We performed a literature review of existing literature, using papers with the highest number of keywords in common and thematic similarity.

Abstract Bullous pemphigoid (BP) is an autoimmune skin disease predominantly found in elderly people, which causes blistering of the skin and severe itching. The incidence of BP reported by previous studies has varied greatly between 0.05 and 42.8 per 1 million persons per year. Higher incidences have been reported in Western Europe and the USA, while countries around the Mediterranean have reported lower rates. However, the epidemiology of BP has not previously been studied in any Scandinavian country. The one-year mortality of BP is highly variable with estimates between 11% and 41% worldwide. As for comorbidities, the previous studies have shown that BP is associated with neurological disorders. The aim of this study was to investigate the incidence and mortality of BP in Finland, to assess the treatments used for BP, and the potential contribution of systemic glucocorticoid treatment to the high mortality rate found in BP patients. A further aim was to obtain more specific information about the neurological diseases associated with BP, and to clarify the less studied association with psychiatric disorders. For these purposes, we collected the records of all immunologically confirmed BP patients diagnosed in the Oulu University Hospital between 1985 and 2012, and, for a sub-study III, data for all patients diagnosed with BP in Finnish hospitals between 1987 and 2013. We found that the incidence of BP in Northern Finland has increased over the past two decades to approximately 27 new BP cases per 1 million persons per year. The one-year mortality of BP patients is 17%, and the standardized mortality ratio (SMR) is 7.6. Common comorbidities found in the sample of BP patients were: cardiovascular diseases (76%), neurodegenerative diseases (41%), skin conditions other than BP (37%) and type 2 diabetes (23%). Many neurodegenerative diseases of the central nervous system were associated with BP, as were many psychiatric disorders. The association was strongest between multiple sclerosis (MS) and BP, with MS patients having almost a 6-fold higher risk of BP than controls. The present study reports for the first time the incidence and mortality of BP in Finland, and provides new information about the association between BP and neurological and psychiatric disorders
Tiivistelmä Rakkulainen pemfigoidi (josta jatkossa käytetään nimitystä pemfigoidi) on autoimmuunisairaus, joka esiintyy yleensä iäkkäillä, ja aiheuttaa ihon rakkulointia ja hankalaa kutinaa. Aiemmissa tutkimuksissa pemfigoidin ilmaantuvuus on vaihdellut 0,05:sta 42,8:aan tapaukseen miljoonaa ihmistä kohden vuodessa. Ilmaantuvuuden on havaittu olevan korkeampi Länsi-Euroopassa, kun taas Välimeren ympäristössä ilmaantuvuus on matalampi. Pemfigoidia sairastavien kuolleisuus vuoden kuluessa diagnoosista vaihtelee noin 11-41%:n välillä. Aiemmat tutkimukset ovat myös osoittaneet, että pemfigoidi liittyy neurologisiin sairauksiin. Pemfigoidin epidemiologiaa ei ole kuitenkaan tutkittu Suomessa tai muissa Pohjoismaissa. Tämän tutkimuksen tarkoituksena oli selvittää pemfigoidin ilmaantuvuus ja kuolleisuus Suomessa, tutkia sen hoitoon käytettyjä lääkkeitä sekä arvioida systeemisen glukokortikoidihoidon osuutta korkeaan kuolleisuuteen. Lisäksi tavoitteena oli saada yksityiskohtaista tietoa pemfigoidiin liittyvistä neurologisista sairauksista ja selvittää lisää aiemmissa tutkimuksissa ristiriitaiseksi jäänyttä yhteyttä psykiatrisiin sairauksiin. Tätä varten keräsimme tiedot kaikista Oulun yliopistollisessa sairaalassa diagnosoiduista, immunologisesti varmennetuista pemfigoiditapauksista vuosilta 1985-2012. Kolmannessa osatyössä käytimme kansallista aineistoa, joka sisälsi kaikkialla Suomessa diagnosoidut pemfigoidia sairastavat potilaat vuosilta 1987-2013. Pemfigoidin ilmaantuvuus kasvoi seuranta-aikana ollen nykyisin Pohjois-Suomessa noin 27 tapausta miljoonaa ihmistä kohden vuodessa. Kuolleisuus vuoden kuluessa diagnoosista oli 17% ja vakioitu kuolleisuussuhde (standardized mortality ratio) 7,6. Yleisiä oheissairauksia pemfigoidia sairastavilla olivat sydän- ja verisuonisairaudet (76%), neurodegeneratiiviset sairaudet (41%), muut ihosairaudet (37%) sekä tyypin 2 diabetes (23%). Tutkimuksessa todettiin, että monet neurogeneratiiviset sairaudet ja monet psykiatriset sairaudet liittyvät pemfigoidiin. Yhteys oli vahvin pesäkekovettumataudin (MS-tauti) ja pemfigoidin välillä, ja MS-tautia sairastavilla riski sairastua pemfigoidiin oli lähes 6-kertainen verrattuna kontrollipotilaisiin. Tämä tutkimus on ensimmäinen, joka raportoi pemfigoidin ilmaantuvuuden ja kuolleisuuden Suomessa. Tutkimus antaa lisäksi uutta tietoa pemfigoidin yhteydestä neurologisiin ja psykiatrisiin sairauksiin

Лікувальне використання низьких температур відоме з давніх давен. Терапевтичний ефект місцевої дії холоду для зупинки кровотеч і при травматичному шоці описував ще Гіппократ. В XX ст. розвиток науково-технічного прогресу дав можливість використовувати низькі температури з лікувальною метою.

Abstract Glucocorticoids are the most important and widely used treatment modality in dermatology. A large variety of topical as well as systemic preparations is available. Most patients treated with glucocorticoids respond quickly to the treatment, but some are considered insensitive or even resistant to glucocorticoid therapy. Currently, there is no known measurable variable, through which the response can be predicted. Glucocorticoids mediate their actions through glucocorticoid receptors (GR). Several isoforms of GR exist, but the α (GRα) and β (GRβ) isoforms are clinically the most important. Based on previous studies, it has been proposed that the abundance of GR isoforms or the GRβ: GRα –ratio could affect individual responsiveness to corticosteroid treatment. In particular, up-regulation of GRβ expression has been shown to be linked to resistance to corticosteroid treatment. This thesis comprises three sub-studies. Firstly, we wanted to determine whether GRα and GRβ are expressed in inflammatory skin diseases. Secondly, we examined if the expression is altered by corticosteroid treatment in eczema atopicum, bullous pemphigoid and psoriasis. Finally, we measured the effects of a topical vitamin D3 analogue (calcipotriol) combined with betamethasone compared with betamethasone monotherapy on inflammatory biomarkers of psoriasis. Our studies provide detailed novel data about the expression of GRα and GRβ. GRα and GRβ were shown to be expressed in the blood lymphocytes and lesional skin of patients with eczema atopicum, bullous pemphigoid and psoriasis, as well as in the skin of patients with eczema nummulare, lichen simplex chronicus and lichen ruber planus. Systemic corticosteroid treatment was shown to affect the expression of GRα and GRβ in eczema atopicum and bullous pemphigoid, but the inconsistent variation in their expression between patients prevented us from drawing firm conclusions. Neither GRα nor GRβ as a single marker were found to be a suitable predictor of corticosteroid responsiveness. Clinical and laboratory analyses showed that topical treatment of psoriasis with calcipotriol/betamethasone combination ointment is more beneficial measured by both than betamethasone monotherapy
Tiivistelmä Glukokortikoideja (”kortisoni”) käytetään tulehduksellisten ihotautien hoidossa paikallisesti tai systeemisenä lääkkeenä. Suurin osa potilaista reagoi hoitoon nopeasti, mutta osalla hoitovaste on heikompi tai ilmenee hitaasti. Tällä hetkellä ei tunneta keinoja ennustaa luotettavasti kortisonihoidon vastetta. Glukokortikoidit vaikuttavat elimistössä glukokortikoidireseptorien (GR) kautta. Glukokortikoidireseptorista tunnetaan useita alatyyppejä, joista tärkeimmät ovat α (GRα) ja β (GRβ). Aiemman tiedon pohjalta on pidetty mahdollisena, että GR-alatyyppien suhteella tai määrällä on merkitystä kortisonivasteen syntymisessä. Erityisesti on arveltu, että ylimäärä GRβ:aa voisi estää kortisonihoidon vaikutusta. Tässä väitöskirjassa tavoitteena on ollut selvittää, tapahtuuko GR-alatyyppien ilmenemisessä muutoksia tulehduksellisia ihosairauksia sairastavilla potilailla sekä tutkia, miten kortisonihoito vaikuttaa GR-tasoihin atooppista ihottumaa, pemfigoidia ja psoriaasia sairastavilla potilailla. Lisäksi olemme verranneet paikallishoitoa pelkällä kortisonivoiteella D-vitamiinijohdos kalsipotriolin ja kortisonin yhdistelmähoitoon psoriaatikoilla. Tutkimus on antanut uutta yksityiskohtaista tietoa GRα:n ja GRβ:n esiintymisestä ihossa ja tulehdussoluissa ihosairauksia sairastavilla potilailla. Tutkimustulosten perusteella voidaan todeta, että GRα ja GRβ esiintyvät atooppista ihottumaa, pemfigoidia ja psoriaasia sairastavien potilaiden ihossa ja veren tulehdussoluissa sekä nummulaari-ihottumaa, neurodermatiittia ja punajäkälää sairastavien potilaiden ihossa. Suun kautta annettu kortisonihoito vaikuttaa GRα- ja GRβ–lähetti-RNA:n ilmenemiseen, mutta potilaskohtaiset erot ovat suuret, eikä kumpikaan, GRα tai GRβ, sovellu yksinään ennustamaan kortisonihoidon vastetta. Paikallisella kortisonihoidolla D-vitamiinijohdos kalsipotrioliin yhdistettynä on suotuisampi vaikutus psoriaasin tulehduksellisiin välittäjäaineisiin ja tulehdussoluihin kuin pelkällä paikallisella kortisonihoidolla

Apurinic/apyrimidinic DNA repair endonuclease-1 (APE1), first recognized as an important DNA excision repair enzyme, is also known as Redox Factor-1 (Ref-1) involved in the activation of many nuclear transcription factors in both redox-dependent and independent manner. It has been well-documented that the overexpression of APE/Ref-1 contributes to the development of chemo-resistance and is associated with tumor progression in many human malignancies [1]. Our previous study in melanoma demonstrated that the development of novel inhibitors targeting the redox regulation domain of APE/Ref-1 is a promising strategy for melanoma treatment. To date, limited successes have been reported in developing novel APE/Ref-1 inhibitors for cancer treatment. Utilizing a structure-based approach, our study identified and characterized small molecular inhibitors of APE/Ref-1. First, N-terminally truncated APE/Ref-1 protein lacking the first 40 amino acid residues (∆40APE-1wt) was cloned into the pGEX-6P1 vector to express the GST-∆40APE-1wtprotein. After cleavage of GST-tag, the concentrated ∆40APE-1wt protein was subjected to protein crystallization study. We have successfully diffracted ∆40APE-1wt crystals and collected data with a resolution of 1.57Å. The crystal structure was further determined by molecular replacement in Molrep using the already available human APE-1 structure (PDB: 5CFG). For the first time, we observed the dimerization of APE/Ref-1 protein formed under oxidative conditions, which may contribute to the redox regulation of APE/Ref-1. Such structural transformation of APE/Ref-1 protein under distinct redox conditions may pave the way for future drug development and optimization. The binding affinity of the candidate compounds with ∆40APE-1wt protein was also determined using Surface Plasmon Resonance (SPR), and the Ki values were analyzed. One of the potent inhibitors developed by our group by structure-based approach, exhibited promising anti-melanoma activities both in vitro and in vivo. Future studies on the structure-activity association are warranted.

Remarkable progress has been made in the development of effective treatments for patients with rheumatoid arthritis (RA). To ensure that a patient is optimally responding to treatment, consistent monitoring of disease activity is recommended. Established composite and individual disease activity measures often cannot be computed due to missing laboratory values. Simplified measures that can be calculated without a lab value have been developed and previous studies have validated these new measures, yet differences in their performance compared with established measures remain. Therefore, the goal of my doctoral research was to examine and evaluate disease activity and composite measures to facilitate monitoring of response in clinical care settings and inclusion of patients with missing laboratory values in epidemiological research. In the first study, the validity of two composite measures, the Clinical Disease Activity Index (CDAI) and the Disease Activity Score with 28 joint count (DAS28) was examined and both were significantly associated with a rheumatologist’s decision to change therapy (CDAI OR=1.58; 95% CI: 1.42, 1.76) (DAS28 OR=1.34; 95% CI 1.27,1.56). However, further evaluation using receiver operating characteristic (ROC) analysis found that they were not strong predictors of physician decisions to change therapy (AUC=0.75, 0.76, respectively). Thus, they should not be used to guide treatment decisions in the clinic. Two measures of disease activity, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) are often not measured and impede the computation of composite measures of disease activity. In the second study, significant factors which may predict the measurement of the ESR and CRP were identified and included physician and clinical variables but no quantitative disease activity measures. Thus the suitability of the ESR and CRP as measures of disease activity is suspect. In the final study, I created a new composite measure, the modified disease activity score with 28 joint count (mDAS28), by replacing the laboratory value in the DAS28. The mDAS28 was then validated by comparing its performance with the DAS28. The measures were strongly correlated (r=0.87), and strong agreement was found between the two measures when categorizing patients to levels of disease activity (ĸ=0.77) and treatment response (ĸ=0.73). Therefore, the mDAS28 could be used in place of the DAS28 when laboratory values needed to compute the DAS28 are missing. In summary, I found that the CDAI and DAS28 were not strong predictors of the rheumatologist’s decision to change therapy. I also found that the variability in the measurement of ESR and CRP was not associated with disease activity. I was able to modify the DAS28 by replacing the laboratory measure and create a new simplified measure, the mDAS28. I also validated the mDAS28 for use in the clinic and in epidemiological research when the DAS28 is unavailable.

The skin is the largest organ of the human body and is responsible for numerous vital functions as being a physical protective barrier against the invasion of pathogenic microorganisms, penetration of harmful chemicals, and of ultraviolet rays, while having also important roles as in body fluid homeostasis and the elaboration of immune and sensitive responses. The cooperation between the various constituting elements of the skin (keratinocytes, fibroblasts, immune system cells, motor and sensitive nerve fibres, glands, blood and lymphatic vessels) together with the microorganisms residing on its surface is important to keep the body healthy. Numerous common skin disorders such as atopic dermatitis, psoriasis and acne can be associated with an individual's genetic predisposition, but also with an imbalance in the microflora of the skin or even to an uncontrolled activation of responses of the innate and adaptive immune system due often to over-expression of proteins such as cyclooxygenase-2 (COX-2), responsible for the genesis of chronic inflammatory processes and oxidative stress that lead to the production of molecules such as nitric oxide (NO) and reactive oxygen species (ROS) that, when in excess, become toxic and harmful to the cells. Humulus lupulus Linnaeus, belonging to the Cannabaceae family, is popularly used in beer making and in traditional medicine for its relaxing therapeutic properties, such as the treatment of insomnia and anxiety. The inflorescence of H.lupulus is the part of the plant mostly used because it is where the lupulin gland is located, an organ harbouring mainly 15-30% of resins (hard and soft resins), essential oils, polyphenols, among other minority compounds responsible for the medicinal character of the plant. These secondary metabolites of H.lupulus are known to have a high anti-inflammatory, anxiolytic, antidepressant, antioxidant and antimicrobial potential. In this dissertation, the capacity of aqueous extracts of the flower, the mix aqueous extract (mixture of stems, leaves and flowers), the flower hydrolate and the mix hydrolate of H.lupulus was studied to decrease multiplication of Gram-positive (Staphylococcus aureus, Staphylococcus epidermidis and Cutibacterium acnes) and Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria by the microdilution method and in the alteration of the metabolic activity of 3T3 fibroblast cells and RAW macrophages from mouse skin by the colorimetric method, 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT). The impact of these extracts on the production of NO was also evaluated by the Griess method, production of ROS by fluorescent molecules such as H2DCFDA, and on the expression of COX-2 an agent involved in the inflammation processes after activation of macrophages by lipopolysaccharide (LPS) by Western blot. The results show that H.lupulus flower hydrolate presents higher antibacterial capacity (especially against Gram-positive bacteria), strong antioxidant capacity, because it significantly reduces the production of ROS reduces inflammation, decreasing the production of NO and decreasing the expression of COX-2 by LPS activated macrophages. The remaining extracts also presented anti-inflammatory and antioxidant effects, but less evident than the flower hydrolate. Mix hydrolate and aqueous extract showed a low antibacterial effect for some of the strains under study and did not significantly alter the metabolic activity of 3T3 fibroblasts and RAW macrophages. This study demonstrated that H.lupulus, mainly the inflorescence of the plant (flowers), has chemical elements with anti-inflammatory, antioxidant and antibacterial capacities supporting the concept of being H.lupulus products very interesting to be used in treatment and prevention of diseases of the skin while respecting its physiological constitution . However, further studies are necessary in order to better understand the profile of the different chemical compounds of H.lupulus and to establish the best effective concentration yet non-toxic for the cells and microbiota of skin.
A pele, é o maior órgão do corpo humano e é responsável por inúmeras funções vitais como barreira física protetora contra a invasão de microrganismos patogénicos, penetração de produtos químicos nocivos e de raios ultravioleta, mas também desempenha papéis importantes na homeostasia corporal e na elaboração de respostas imunes e sensitivas. A cooperação existente entre os vários elementos constitutivos da pele (queratinócitos, fibroblastos, células do sistema imunitário, fibras nervosas motoras e sensitivas, glândulas, vasos sanguíneos e linfáticos) e até mesmo com os microrganismos, da sua superfície é importante para manter o organismo saudável. Inúmeros distúrbios frequentes da pele como a dermatite atópica, a psoríase e a acne podem estar associadas a uma predisposição genética do indivíduo, mas também a um desequilíbrio da microflora da pele ou mesmo a uma ativação sem controlo de respostas do sistema imunitário inato e adaptativa devido muitas vezes a sobrexpressão de proteínas tais como a ciclooxigenase-2 (COX-2) responsáveis pela génese de processos inflamatórios e de stress oxidativo crónicos que levam à produção de moléculas como o óxido nítrico (NO) e espécies reativas de oxigénio (ROS) que quando em excesso se tornam tóxicos e deletérios para as células. O Humulus lupulus Linnaeus, pertencente a família dos Cannabaceae é popularmente utilizado na confeção da cerveja e na medicina tradicional pelas suas propriedades terapêuticas relaxantes, como, por exemplo, no tratamento de casos de insónia e ansiedade. A inflorescência do H.lupulus é a parte da planta mais utilizada por ser onde está localizada a glândula da lupulina, órgão constituído maioritariamente por resinas 15- 30% (resinas duras e macias), óleos essenciais, polifenóis, entre outros compostos minoritários responsáveis pelo carater medicinal da planta. Estes metabolitos secundários do H.lupulus são conhecidos por apresentarem um alto potencial anti-inflamatório, ansiolítico, antidepressivo, antioxidante e antimicrobiano. Na presente dissertação, foi estudada a capacidade de extratos aquosos da flor, do mix (mistura de caules, folhas e flores), do hidrolato da flor e do hidrolato do mix do H.lupulus na diminuição da multiplicação de estirpes bacterianas Gram-positivas (Staphylococcus aureus, Staphylococcus epidermidis e Cutibacterium acnes) e Gram-negativas (Escherichia coli e Pseudomonas aeruginosa) pelo método de microdiluição e na alteração da atividade metabólica de células de fibroblastos 3T3 e de macrófagos RAW provenientes da pele de ratinhos através do método colorimétrico, brometo de [3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium] (MTT). Foi também avaliado o impacto destas substâncias na produção de NO, através do método de Griess, produção de ROS através de moléculas fluorescentes como o H2DCFDA, e na expressão da COX-2, agente interveniente nos processos de inflamação após ativação dos macrófagos pelo lipopolissacarideo (LPS), por Western blot. Os resultados obtidos demonstram que o hidrolato da flor do H.lupulus apresenta maiores capacidades antibacterianas, especialmente contra as bactérias Gram-positivas, forte capacidade antioxidante, porque diminui significativamente a produção de ROS, reduz a inflamação, diminuindo a produção de NO e a expressão da COX-2 pelos macrófagos ativados pelo LPS. Os restantes extratos apresentaram também efeito anti-inflamatório e antioxidante, mas menos evidente do que o hidrolato da flor. O hidrolato do mix e o extrato aquoso do mix demonstraram um baixo efeito antibacteriano para algumas das estirpes em estudo e não alteraram significativamente a atividade metabólica dos fibroblastos 3T3 e dos macrófagos RAW. Este estudo demonstrou que o H.lupulus, principalmente a inflorescência da planta (flores) possui elementos químicos com capacidades anti-inflamatórias, antioxidantes e antibacterianas, podendo ser bastante interessantes na conceção de produtos à base de lúpulo com o intuito terapêutico e de prevenção de doenças que possam surgir na pele respeitando ao mesmo tempo a sua constituição fisiológica. Contudo mais estudos são necessários de modo a se conhecer melhor o perfil dos diferentes compostos químicos do H.lupulus bem como de modo a se estabelecer concentrações não toxicas para as células e microrganismos que compõem a microbiota da pele.

博士
國立成功大學
化學工程學系
106
This study focuses on the feasibility of transdermal delivery of drug particles using dissolvable polymer microneedles and its potential for the treatment of skin diseases. Investigating the efficiency of transdermal delivery of poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles using polyvinyl alcohol/polyvinyl pyrrolidone (PVA/PVP) microneedles is the first section. Transdermal delivery of doxorubicin (DOX) and IR 780-loaded polycaprolactone (PCL) microparticles using hyaluronic acid microneedles for treating A431 tumor by combination of chemotherapy, photodynamic and photothermal therapy is the second section. The topic is efficient delivery of nanoparticles to deep skin layers using dissolvable microneedles with an extended-length design in the first section. Skin pretreatment with microneedles (MNs) increases drug permeation through the skin by creating microchannels in the skin. However, because of skin’s inherent elasticity and self-healing ability, these microchannels shrink or reseal rapidly, thus limiting the nanoparticle (NP) delivery efficiency. This study evaluated dissolvable PVA/PVP MNs with an extended-length design for the efficient transdermal delivery of NPs. In this system, PLGA NPs are encapsulated within the pyramidal structure of the MNs. The extended length of the PVA/PVP MN allows it to counteract skin indentation during insertion, thus enabling complete insertion of the pyramidal structure into the skin to deliver the NPs. In contrast to MN pretreatments that require passive diffusion of NPs through the skin, the extended PVA/PVP MNs can directly bring the NPs into the deeper skin layers, and then rapidly dissolve in 3 min to release the payload. An in vivo transdermal delivery study showed that approximately 90% of the loaded NPs were delivered to the viable epidermis and dermis, whereas only 2% of topically applied NPs were detected in the skin after being treated with a commercial MN product (3MTM Microchannel Skin System). The NPs delivered by the extended MN remained at the insertion site for 5 days, thus providing sustained drug release. The proposed MN system could be a promising tool for the transdermal delivery of NPs for treating deep skin diseases such as bacterial infection and malignant tumor. The topic is delivery of IR 780 and DOX-loaded microparticles into skin cancer using hyaluronic acid microneedles with an extended-length design for treating squamous cell carcinoma in the second section. IR 780 is a near infrared (NIR) heptamethine indocyanine dye, it can exhibite remarkable photothermal effect and photodynamic therapy with NIR light. The NIR light-triggered release experiments demonstrated that PCL microparticles can quickly melt at 48˚C due to the heating of the encapsulated IR 780 when NIR irradiation and then release DOX into the medium. In vitro cell viability assays showed that IR 780 and DOX-loaded PCL microparticles significantly enhanced the cytotoxicity of combination therapy in A-431 cells. Dramatic size decreases in squamous cell carcinoma xenografts were observed for delivery of IR 780 and DOX-loaded PCL microparticles into skin tumor using dissolvable hyaluronic acid microneedles in ICR SCID mice. Compared with IR 780-loaded PCL microparticles, the combined treatment with chemotherapy, photodynamic and photothermal therapy possessed a better therapeutic efficacy, resulting from a DOX effect. In conclusion, we believe that delivery of DOX and IR 780-loaded microparticles into tumor using dissolvable microneedles are promising and may serve as a useful system for superficial cancer treatment.