Background: Patients with nonallergic irritant rhinitis (NAIR) have symptoms of nasal congestion, nasal irritation, rhinorrhea, and sneezing in response to nasal irritants. We currently have no reliable objective means to quantify these patients' subjective symptoms. In this study, we used the transient receptor potential vanilloid receptor (TRPV1) receptor agonist, capsaicin, as an intranasal challenge while comparing the changes in blood flow with optical rhinometry between subjects with NAIR and healthy controls (HCs). Methods: Six HCs and 6 NAIR subjects were challenged intranasally with saline solution followed by increasing concentrations of capsaicin (0.005 mM, 0.05 mM, and 0.5 mM) at 15-minute intervals. We recorded maximum optical density (OD) and numeric analog scores (NAS) for nasal congestion, nasal irritation, rhinorrhea, and sneezing for each subject after each challenge. Correlations between NAS and maximum OD were calculated. Results: Maximum OD increased with increasing concentrations of intranasal capsaicin in NAIR subjects. There were significant differences in maximum OD obtained for 0.05 mM and 0.5 mM capsaicin between NAIR subjects and HCs. Significant differences were found in the NAS for nasal irritation at 0.005 mM, 0.05 mM, and 0.5 mM, and nasal congestion at 0.5 mM. Correlation between maximum OD and mean NAS was most significant for 0.05 mM capsaicin. Conclusion: Optical rhinometry with intranasal capsaicin challenge could prove a viable option in the diagnosis of NAIR. Further studies will investigate its use to monitor a patient's response to pharmacologic therapy and provide further information about the underlying mechanisms of NAIR.

Anyone who has eaten spicy foods has experienced the adverse effects of capsaicin, the pungent chemical found in hot chili that causes a burning sensation. The specific action of capsaicin occurs by the activation of receptors in sensory neurons. This thesis investigates the interaction of capsaicin with model cell membranes representing the structure of neurons. In particular, we are interested in the changes induced by capsaicin to the structure and dynamics of membranes. Molecular dynamics simulations are used to study the molecular interactions. The first part of this study evaluates different molecular representations for capsaicin in an 1-octanol/water system. This inhomogeneous system is commonly used to determine the partition of compounds between hydrophilic and hydrophobic environments, as that found in biological membranes. The results of these simulations validate the OPLS united-atom force field as a reasonable molecular representation of capsaicin, as it describes the behavior of capsaicin both quantitatively and qualitatively in 1-octanol/water mixtures. In the second part, simulations are performed for capsaicin and model cell membranes consisting of dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylethanolamine, two of the most commonly found lipids. Simulations investigated capsaicin in the aqueous and lipid phases. The results provide insight into the changes to the bilayers caused by capsaicin. Bilayers containing dipalmitoylphosphatidylethanolamine showed lower permeabilities to capsaicin than those composed of pure dipalmitoylphosphatidylcholine. Temperature is found to be an important factor in the permeability of capsaicin in the bilayer. Capsaicin in the bilayer concentrated in a region beneath the lipid/water interface, in which favorable hydrophilic and lipophilic interactions occur. The structure of the bilayer is not significantly changed at the concentrations of capsaicin considered. One important result from the simulations indicates that the interfacial density decreases with increasing capsaicin concentration in the bilayer, supporting the experimental observations of increased permeability in bilayers exposed to capsaicin. / Master of Science

Die Osteoarthrose (OA) des Ellbogengelenks ist eine der wichtigsten Gelenkerkrankungen des Hundes, die sich aufgrund erblich bedingter Fehlstellung der Gelenke sekundär häufig schon bei sehr jungen Hunden entwickelt und zu einer verminderten Lebensqualität der Tiere führt. Im Laufe der Erkrankung kommt es zu einer Schädigung des Gelenkknorpels, der an Elastizität und Spannkraft verliert und seine Funktion, nicht mehr ausüben kann. Eine Ablösung von Knorpelfragmenten und Entzündungen im Gelenk können die Zerstörung des Gelenkknorpels beschleunigen. Die OA ist nicht heilbar. Das Therapieziel ist eine möglichst vollständige Schmerzreduktion und die Verbesserung der Lebensqualität. Eine chirurgische Denervation ist im Ellbogengelenk des Hundes ohne Traumatisierung anatomischer Strukturen nicht möglich. Ziel der vorliegenden Arbeit war es, erstmalig einen experimentellen Ansatz zur chemischen Denervation sensibler Gelenkafferenzen im Ellbogengelenk beim Beagle mit dem Neurotoxin OX7-Saporin durchzuführen. Zusätzlich wurden Substanzen aus der Humanmedizin zur Schmerzreduktion und symptomatischen Therapie der OA am Hund erprobt und der Therapieerfolg evaluiert. Im Rahmen eines Therapieversuches in der Klinik für Kleintiere der Universität Leipzig wurde bei zwei, an OA erkrankten Hunden Capsaicin einmalig intraartikulär injiziert. Der erste Hund erhielt intraartikulär 250 mg Capsaicin. Dieser Patient zeigte Nebenwirkungen in Form einer Herz- und Atemfrequenzerhöhung, starker Schmerzhaftigkeit und neurologischen Symptomen. Der zweite Hund wurde mit 83 mg Capsaicin behandelt und zeigte keine Nebenwirkungen. Sowohl die Bewertung der Besitzer als auch die orthopädische Untersuchung ergaben keine Verbesserung der Lahmheit. Vier kranken Hunden wurde ebenfalls im Rahmen eines Therapieversuches Botox (Botulinumtoxin A) intraartikulär einmalig injiziert. Drei Hunde wurden mit 50 Einheiten, ein Hund mit 100 Einheiten Botox behandelt. Die Applikation von Capsaicin und Botox führten zu keiner Verbesserung der Symptomatik. In der Humanmedizin ist die Schmerzbehandlung mit Capsaicin und Botox erfolgreich. Deshalb sollten diese Therapieansätze auch für die Anwendung beim Hund weiter optimiert werden. Das Neurotoxin OX7-Saporin wurde in einer Dosierung von 100 µg erstmalig in das Ellbogengelenk von drei Beagle-Hündinnen appliziert, um eine sensible Denervation des Gelenkes zu erreichen. Der retrograde Transport des Ribosomen-inaktivierenden Proteins in die Perikarya der Spinalganglienzellen, die das Ellbogengelenk sensibel innervieren, sollte eine selektive Zerstörung der Neurone durch das Neurotoxin bewirken. Der retrograde Tracer Fluoro-Gold wurde, zur Kontrolle der Wirkung des Neurotoxins, nach einer Wartezeit von 15 Tagen intraartikulär in das linke und das rechte Ellbogengelenk appliziert. Fluoro-Gold kann ausschließlich von intakten Nervenfasern transportiert werden. Die Ganglien C4 - Th3 wurden bilateral zur histologischen Auswertung entnommen und in Paraffin eingebettet. Zur Bestimmung der Gesamtneuronenanzahl wurden alle Neurone in jedem dritten Schnitt eines Ganglions gezählt. Die Summe dieser Nervenzellen ergab die Gesamtneuronenanzahl eines Ganglions. Die Einlagerung von Lipofuszin führte zu einer starken Autofluoreszenz im Zytoplasma der Neuronen. Alle Schnitte wurden mit Sudan-Schwarz gefärbt, um die Autofluoreszenz von der retrograden FG-Markierung zu unterscheiden. Die FG-Markierung konnte nur in den Ganglien C6 und Th1 im Zytoplasma sehr weniger Neuronen dokumentiert werden. Der histologische Nachweis einer Neurodegeneration nach Applikation des Neurotoxins OX7-Saporin in das linke Ellbogengelenk war negativ. In den untersuchten Spinalganglien wurden intakte Nervenzellen nachgewiesen. Auch die bilaterale intraartikuläre Injektion des retrograden Tracers FG war nicht erfolgreich. Aus den Ergebnissen dieser experimentellen Ansätze wird zusammenfassend geschlussfolgert: Ø Vor der Durchführung zukünftiger experimenteller Untersuchungen beim Hund muss die optimale Konzentration und die Wartezeit für das Neurotoxins OX7-Saporin ermittelt werden. Ø Die Rezeptoren für OX7-Saporin müssen an den Nervenzellen des Hundes zweifelsfrei nachgewiesen werden, bevor das Neurotoxin im caninen Tiermodell eingesetzt werden kann. Ø Die optimale Konzentration des Fluoreszenzfarbstoffes FG für intraartikuläre Injektionen und dessen retrograde Transportgeschwindigkeit beim Hund müssen ermittelt werden, bevor dieser Tracer wieder im Tierexperiment eingesetzt werden kann. Ø FG wurde als 1,8%ige Lösung in das Ellbogengelenk beim Beagle appliziert. Diese Konzentration war wahrscheinlich zu gering, um eine retrograde Markierung der sensiblen Neurone in den entsprechenden Spinalganglienzellen nachzuweisen. Ø Die intraartikuläre Applikation des Neurotoxins OX7-Saporin führte nicht zur selektiven Neurodegeneration. Eine Aussage zur Schmerzausschaltung und symptomatischen Therapie der OA des Hundes mit OX7-Saporin ist deshalb nicht möglich.

Transient receptor potential (TRP) channels are expressed throughout the central nervous system and have a unique ability to detect a wide range of stimuli including changes in voltage, temperature, pH, lipid environment, small molecule agonists, and mechanical stress. While it is known that TRP channels contain the same six transmembrane helix (S1-S6), tetrameric architecture as voltage-gated channels, the degree to which functional and structural analogies are relevant remains poorly understood. This thesis describes a multidisciplinary approach toward understanding the structure and function of TRP channel transmembrane domains by focusing on the S1-S4 transmembrane helices of the TRPV1. This focus is inspired by the voltage-sensor domain (VSD) of the S1-S4 helices of voltage-gated channels, for which a range of studies show functional and structural independence. While some TRP channels are voltage-sensitive, their S4 helix does not contain the positive string of amino acids of canonical VSDs. However, the S1-S4 helices are functionally significant as the binding site of small molecule ligands in both TRPV1 and TRPM8 (for capsaicin and menthol, respectively). The question of TRP channel transmembrane domain modularity is addressed in this thesis by expression and purification trials as well as radioligand-binding assays. It is demonstrated that the S1-S4 and S1-S6 helices of TRPV1 can be properly inserted, overexpressed, and show signs of stability upon detergent-extraction from Saccharomyces cerevisiae membranes. However the TRPV1 S1-S4 and S1-S6 helices do not show wildtype (WT)-like binding in [³H]-RTX binding assays. These results indicate that the TRPV1 transmembrane domains are likely structural but not functional domains. The S. cerevisiae expression system remained promising for the overexpression of TRP transmembrane domains as well as the production of functional, though not stable upon detergent-extraction, WT TRPV1. This WT TRPV1 was subsequently found to functionally bind both RTX, used in ligand binding assays, as well as the double-knot toxin (DkTx), targeted to the pore domain (the S5-S6 helices). An effect of DkTx on RTX binding affinity demonstrates an allosteric interaction indicative of a possible tighter packing between the two transmembrane domains than is seen in voltage-gated channels containing the canonical VSD. Computational approaches additionally allowed for the investigation of the intramembrane capsaicin binding site in the TRPV1 S1-S4 helices, crucial to the initial motivations of this study. While the literature locates the capsaicin binding site to the TRPV1 S1-S4 helices, a `binding pocket' has yet to be defined, with regards to the orientation of bound capsaicin and its access route to the site via the bilayer. Using molecular dynamics (MD) simulations the preferred location of capsaicin within the bilayer is defined, as well as the elucidiation of capsaicin flip-flop between bilayer leaflets as a key event prior to TRPV1 binding. A transient binding was also observed between a homology model of the TRPV1 S1-S4 helices and capsaicin, possibly encouraging the idea that the S1-S4 helices still contain a partial binding site, though of too low affinity to be observed in the binding experiments performed here.

O câncer é o segundo grupo de doenças que mais mata em todo o mundo, atrás apenas das doenças cardiovasculares. No Brasil, esse panorama é preocupante devido ao período de acentuada tendência de envelhecimento, fase etária de maior incidência do câncer. Neste contexto, a atual terapia antineoplásica continua apresentando diversos vieses quanto à toxicidade e seletividade. Muitos dos fármacos empregados na clínica médica são de origem natural, ou tiveram em suas etapas iniciais de desenvolvimento, correlação com produtos naturais, enaltecendo a importância de fontes naturais no desenvolvimento de novos compostos bioativos. A capsaicina, substância responsável pela pungência apresentada pelos frutos de pimenteiras do gênero Capsicum, tem se mostrado um importante agente natural com atividade citotóxica à diversas linhagens neoplásicas, porém, sua ação pungente e baixa estabilidade físico-química, limitam seu emprego terapêutico. Com isso, a capsaicina apresenta grande potencial em servir como arcabouço para o planejamento de novas entidades químicas bioativas, análogas à mesma, para emprego no tratamento do câncer. Este trabalho propôs-se a planejar e sintetizar análogos sulfonamídicos e sulfonatos da capsaicina, promovendo variações moleculares no núcleo estrutural capsaicinóide. Após síntese em única etapa, os análogos foram obtidos e caracterizados por faixa de fusão, RMN 1H/13C e análise elementar. Posteriormente os compostos foram submetidos a ensaio de citotoxicidade nas linhagens tumorais: MDA-MB-231 e MCF-7 (de câncer de mama), B16-F10 (melanoma murino) e sobre linhagens sadias de fibroblasto (3T3), pelo método de avaliação da viabilidade celular por biorredução do sal de tetrazólio, o MTT. Quatro compostos apresentaram atividade biológica interessante, em concentrações micromolares (µM) iferiores à atividade apresentada pela capsaicina, destacando-se os compostos RPF-101 e RPF-151, os mais ativos e que tiveram seus mecanismos de indução de morte investigados. Estudos teóricos subsequentes realizados com os análogos de maior atividade (RPF-101 e RPF-151), revelaram que as modificações moleculares promovidas nas estruturas dos mesmos conferiram maior caráter hidrofílico e maior momento de dipolo, o que pode estar relacionado com o incremento na atividade biológica de ambos. A avaliação dos compostos ativos frente às propriedades constantes na Regra dos Cinco de Lipinski (RO5) revelou que estes respeitam todos os parâmetros da regra, enaltecendo assim o caráter druglikeness dos mesmos, em serem ativos via administração oral, visto que o RPF151 não apresentou ação pungente in vivo. A obtenção dos compostos RPF-101 e RPF-151 indicam o sucesso alcançado com a otimização estrutural promovida no arcabouço capsaicinóide, e enaltece o potencial destes análogos em inspirar o planejamento de novas estruturas, ainda mais otimizadas, que possuam maior potência e menor toxicidade associada. / Cancer is the second most lethal group of diseases all over the world, just behind cardiovascular diseases. This scenery is worrying in Brazil due the pronounced aging trend observed, that presents the higher incidence of cancer. In this context, the current anticancer therapy still have several biases regarding toxicity and selectivity. Many current medicines correlate to natural products, even in its early stages of development, which highlight the importance of natural sources in the design of new bioactive compounds. Capsaicin is the substance responsible for the pungency of Capsicum genus\' chili peppers. Capsaicin has been shown as an important natural agent because of its cytotoxic activity against several tumor cell lines. However, the pungency and poor stability presented by capsaicin restricts its therapeutic employments. Thus, the capsaicinoid scaffold has a great potential to inspire the design of new bioactive analog entities to be employed in anticancer therapy. This project aimed at designing and synthesizing sulfonamide and sulfonate analogues of capsaicin, performing molecular changes on capsaicinoid core. After a single step synthesis, the analogues were purified and characterized by melting point, 1H/13C NMR and elementary analysis. Subsequently, cytotoxicity assays were performed by MTT method on tumor cell lines MDA-MB-231 and MCF-7 (breast cancer), B16-F10 (murine melanoma) and on healthy fibroblast (3T3). Four compounds showed interesting biological activity at micromolar concentrations (µM) and their IC50 were lower than that presented by capsaicin. The most active compounds, RPF-101 and RPF-151, had their death induction mechanisms investigated. In silico subsequent studies with the most active compounds (RPF-101 and RPF-151) showed that molecular changes promoted on each one increased their hydrophilicity and raised their dipole moment, which may be related to the improvement on biological activity of both. The evaluation of active compounds for Lipinski´s Rule of Five (RO5) properties revealed that they respect all properties presenting its druglikeness for oral administration since RPF151 did not presented in vivo pungency. The RPF-101 and RPF-151 indicate the success obtained by the structural optimization promoted on capsaicinoid scaffold and emphasizes the potential of these analogues to inspire the designing of new future optimized structures, with greater potency and less associated toxicity.

Foram desenvolvidos três experimentos para avaliação de uma mistura de extratos vegetais de orégano, pimenta e canela na produção de frangos de corte: Experimento I - Utilizou-se 320 pintos machos de um dia de idade da linhagem Cobb 500, distribuídos em delineamento experimental inteiramente casualisado com quatro tratamentos (0, 75, 150 e 225 ppm de extratos vegetais) com dez repetições de oito aves cada, criadas até 21 dias para determinação da digestibilidade dos nutrientes das dietas, pela metodologia de coleta do conteúdo ileal. Experimento II - Utilizou-se 600 pintos machos de um dia de idade da linhagem Cobb 500, distribuídos em delineamento experimental inteiramente casualisado com cinco tratamentos (Controle Positivo (CP) com 54 ppm de Bacitracina de Zinco; Controle Negativo (CN) sem aditivos melhoradores de desempenho; CN com a adição de 75 ppm; 150 ppm e 225 ppm de extratos vegetais) com dez repetições de doze aves cada para avaliação de desempenho e características de carcaça. Experimento III - Utilizou-se 600 pintos machos de um dia de idade da linhagem Cobb 500, distribuídos em delineamento experimental inteiramente casualisado com cinco tratamentos (Controle Positivo (CP) com 54 ppm de Bacitracina de Zinco; Controle Negativo (CN) sem aditivos melhoradores de desempenho; CP até 33 dias e posteriormente substituído pelo CN, no período de 34 a 42 dias de idade, com a adição de 75 ppm; 150 ppm e 225 ppm de extratos vegetais) com dez repetições de doze aves cada para avaliação de desempenho e características. Todos os dados foram analisados pelo Statistical Analysis System (SAS Institute Inc., 2008), submetidos à análise de regressão polinomial pelo procedimento GLM ao nível de 5% de significância. No experimento I, não foi observado efeito significativo entre os tratamentos para digestibilidade ileal da energia bruta, e a digestibilidade ileal da proteína bruta e dos aminoácidos apresentou efeito quadrático (P<0,05). No experimento II o consumo de ração não apresentou efeito significativo entre os tratamentos, já para ganho de peso e conversão alimentar foi observado efeito significativo (P<0,05) na comparação entre o antibiótico e os níveis de extratos vegetais adicionados a dieta. No experimento III, não foi observado efeito significativo entre os tratamentos para consumo de ração e ganho de peso, mas a conversão alimentar apresentou efeito significativo (P<0,05) na comparação entre o antibiótico e os níveis de extratos vegetais adicionados a dieta. Nos experimentos II e III, para as características de carcaça não foram observados efeitos significativos (P<0,05) entre os tratamentos. Concluiu-se que a inclusão de uma mistura de extratos vegetais de orégano, pimenta e canela na dieta de frangos de corte em substituição aos aditivos melhoradores de desempenho proporcionou maior digestibilidade ileal da proteína bruta e dos aminoácidos, e de acordo com os níveis praticados, os extratos vegetais não proporcionaram efeitos positivos no desempenho quando comparados com o tratamento com antibiótico, no período total de criação. Entretanto, o fornecimento na fase final apresentou um comportamento diferente, melhorando o rendimento de peito. / We developed three experiments to evaluate a mixture of herbal extracts of oregano, pepper and cinnamon in the production of broilers: Experiment I - was used 320 male chicks from one day old Cobb 500, distributed in completely randomized design with four treatments (0, 75, 150 and 225 ppm of plant extracts) with ten replicates of eight birds each, created up to 21 days to determine the nutrient digestibility of diets, the collection methodology of the ileal content. Experiment II - was used 600 male chicks from one day old Cobb 500, distributed in completely randomized design with five treatments (Positive Control (PC) with 54 ppm Bacitracin Zinc; Negative Control (NC) without enhancing additives performance; CN with the addition of 75 ppm, 150 ppm and 225 ppm of plant extracts) with ten replicates of twelve chicks each to assess performance and carcass characteristics. Experiment III - was used 600 male chicks from one day old Cobb 500, distributed in completely randomized design with five treatments (Positive Control (PC) with 54 ppm Bacitracin Zinc; Negative Control (NC) without enhancing additives performance; CP up to 33 days and subsequently replaced by CN, from 34 to 42 days of age, with the addition of 75 ppm, 150 ppm and 225 ppm plant extracts), with ten replicates of twelve chickens each for performance evaluation and characteristics. All data were analyzed using Statistical Analysis System (SAS Institute Inc., 2008), submitted to polynomial regression analysis by GLM procedure at the 5% level of significance. In the first experiment, no significant effect was observed between treatments for ileal digestibility of raw energy and ileal digestibility of crude protein and amino acids showed a quadratic effect (P <0.05). In experiment II feed intake had no significant effect between treatments, as for weight gain and feed conversion was no significant effect (P <0.05) in the comparison between the antibiotic and the levels of plant extracts added to the diet. In experiment III, no significant effect was observed between treatments for feed intake and weight gain, feed conversion but significant effect (P <0.05) in the comparison between the antibiotic and the levels of plant extracts added to the diet. In experiments II and III, for carcass traits were not significant effects (P <0.05) among treatments. It was concluded that the inclusion of a mixture of herbal extracts of oregano, pepper and cinnamon in the diet of broilers in place of performance enhancing additives provided higher ileal digestibility of crude protein and amino acids, and according to the levels prevailing, vegetable extracts yielded no positive effect on performance when compared with the antibiotic treatment, the total periods. However, the provision in the final phase showed a different behavior by improving the yield of breast.

A Síndrome de Ardência Bucal (SAB) caracteriza - se por sensação de ardor, com ausência de sinais clínicos ou laboratoriais associados. A etiopatogenia é desconhecida, inexistindo protocolo terapêutico satisfatório. O objetivo deste trabalho foi avaliar a eficácia da aplicação tópica de capsaicina, como alternativa terapêutica, em um grupo de pacientes portadores da SAB, além de investigar, nessa população, indicativos de ansiedade e depressão, correlacionando estes últimos aspectos com a resposta à terapêutica aplicada. Constituiu-se um grupo de vinte indivíduos portadores da síndrome, todos de acordo com os termos do consentimento esclarecido. Quinze sujeitos constituíram o grupo teste (GT) e foram tratados com capsaicina, em aplicações diárias, durante três semanas, repetindo-se o ciclo por quatro semanas após uma semana de intervalo. O grupo controle (GC) foi tratado com o creme base utilizado como veículo da capsaicina, durante o mesmo período. Ambos foram controlados após 30 dias do término da medicação. A evolução dos sintomas foi controlada por escala visual de sintomatologia (EVS) e questionário acerca do efeito global percebido (EGP). A intensidade média do sintoma de ardência antes do início dos ciclos de tratamento, mensurado pela EVS, foi de 5,1 (GT) e 4,4 (GC). Ao final da quarta semana o GT mostrou redução dos sintomas (EVS=3,6), enquanto o GC declarou aumento da sintomatologia (EVS=4,8). No GT, entre a quarta e a oitava semana houve redução dos sintomas da ordem de 8,3%, e entre a oitava e a décima segunda semana observou-se aumento de 13,5% da sintomatologia. No GC houve 22.8% de piora (EVS=5,75) entre o início e a décima segunda semana. Ao EGP houve pelo menos algum alívio do sintoma em seis pacientes (40%) do GT e em um paciente do GC (20%). Quatro pacientes (26,6%) reportaram remissão total do sintoma após tratamento com capsaicina e um paciente (20%) do controle. Para três pacientes do GT e dois do GC não houve modificações do sintoma. Houve relato de piora em dois pacientes (13,3%) do GT e um (20%) do GC. Oito pacientes do GT apresentaram alto nível de ansiedade e sete níveis médios. No GC um paciente apresentou nível baixo, três mostraram valores médios e um classificou-se como alto. Ao CES-D valores indicativos de depressão foram registrados por dez pacientes (66,6%) do GT e 40% (02) do GC. Concluímos que a capsaicina apresentou efetividade no controle da sintomatologia da SAB, parecendo haver correlação com a intensidade inicial de sintomas e manutenção do uso do medicamento. Além disso, houve correlação entre alto nível de ansiedade e indicativos de depressão, embora não se tenha percebido influência destes aspectos sobre a resposta terapêutica. / Burning mouth syndrome (BMS) is characterized by an oral burning sensation, with no corresponding clinical signs or laboratory abnormalities. The etiology is unknown, and there was no satisfactory treatment available. The objective of this study was to evaluate the effectiveness of topical use of capsaicin, as an alternative therapy in a group of BMS patients, as well as to correlate anxiety and depression levels to response to the therapy applied. Twenty BMS individuals in accordance to the terms to informed consent comprised the study group. Fifteen subjects were allocated to the test group (TG) and were treated with capsaicin, in daily applications for three weeks, one-week interval and an additional treatment cycle of four weeks. The control group (CG) was treated with the cream base used as a vehicle of capsaicin preparation, during the same period. All patients were examined 30 days after discontinuation of the medication. Results were assessed through a visual analogue scale (VAS) and a questionnaire on the global perceived effect (GPE). The average symptoms intensity before treatment, on EVS, was 5.1 (TG) and 4.4 (CG). At the fourth week control, TG presented reduction on the level of symptoms (EVS = 3.6), while CG presented an increase of symptoms intensity (VAS = 4.8). In the TG, between fourth and eighth week of follow-up, symptoms decreased around 8.3%, and between the eighth and twelfth week there was an increase of 13.5% on symptoms intensity. In the CG it was registered 22.8% of worsening (EVS = 5.75) between the beginning of the study and the twelfth week of control. On GPE assessment, six patients (40%) of TG and one patient of CG (20%), presented some relief of symptoms; four patients TG (26.6%) reported total remission of symptoms after treatment with capsaicin and one patient (20%) of control; three patients of TG and two of the CG remained unaltered. There were reports of worsening in two patients (13.3%) of TG and one (20%) of the CG. Eight patients of TG showed a high level of anxiety and seven moderate levels. In CG one patient presented low level, three showed a moderate level and one was ranked as having a high level of anxiety. CES-D suggested traits of depression in ten patients (66.6%) of TG and 40% (2) of the CG. We concluded that capsaicin is effective in controlling the burning symptom of BMS, suggesting some correlation with initial symptoms intensity and the maintenance of drug use. Moreover, there was some correspondence between high levels of anxiety and traits of depression, but it was not perceived influence of these aspects to the therapeutic response.

O câncer é o segundo grupo de doenças que mais mata em todo o mundo, atrás apenas das doenças cardiovasculares. No Brasil, esse panorama é preocupante devido ao período de acentuada tendência de envelhecimento, fase etária de maior incidência do câncer. Neste contexto, a atual terapia antineoplásica continua apresentando diversos vieses quanto à toxicidade e seletividade. Muitos dos fármacos empregados na clínica médica são de origem natural, ou tiveram em suas etapas iniciais de desenvolvimento, correlação com produtos naturais, enaltecendo a importância de fontes naturais no desenvolvimento de novos compostos bioativos. A capsaicina, substância responsável pela pungência apresentada pelos frutos de pimenteiras do gênero Capsicum, tem se mostrado um importante agente natural com atividade citotóxica à diversas linhagens neoplásicas, porém, sua ação pungente e baixa estabilidade físico-química, limitam seu emprego terapêutico. Com isso, a capsaicina apresenta grande potencial em servir como arcabouço para o planejamento de novas entidades químicas bioativas, análogas à mesma, para emprego no tratamento do câncer. Este trabalho propôs-se a planejar e sintetizar análogos sulfonamídicos e sulfonatos da capsaicina, promovendo variações moleculares no núcleo estrutural capsaicinóide. Após síntese em única etapa, os análogos foram obtidos e caracterizados por faixa de fusão, RMN 1H/13C e análise elementar. Posteriormente os compostos foram submetidos a ensaio de citotoxicidade nas linhagens tumorais: MDA-MB-231 e MCF-7 (de câncer de mama), B16-F10 (melanoma murino) e sobre linhagens sadias de fibroblasto (3T3), pelo método de avaliação da viabilidade celular por biorredução do sal de tetrazólio, o MTT. Quatro compostos apresentaram atividade biológica interessante, em concentrações micromolares (&#181;M) iferiores à atividade apresentada pela capsaicina, destacando-se os compostos RPF-101 e RPF-151, os mais ativos e que tiveram seus mecanismos de indução de morte investigados. Estudos teóricos subsequentes realizados com os análogos de maior atividade (RPF-101 e RPF-151), revelaram que as modificações moleculares promovidas nas estruturas dos mesmos conferiram maior caráter hidrofílico e maior momento de dipolo, o que pode estar relacionado com o incremento na atividade biológica de ambos. A avaliação dos compostos ativos frente às propriedades constantes na Regra dos Cinco de Lipinski (RO5) revelou que estes respeitam todos os parâmetros da regra, enaltecendo assim o caráter druglikeness dos mesmos, em serem ativos via administração oral, visto que o RPF151 não apresentou ação pungente in vivo. A obtenção dos compostos RPF-101 e RPF-151 indicam o sucesso alcançado com a otimização estrutural promovida no arcabouço capsaicinóide, e enaltece o potencial destes análogos em inspirar o planejamento de novas estruturas, ainda mais otimizadas, que possuam maior potência e menor toxicidade associada. / Cancer is the second most lethal group of diseases all over the world, just behind cardiovascular diseases. This scenery is worrying in Brazil due the pronounced aging trend observed, that presents the higher incidence of cancer. In this context, the current anticancer therapy still have several biases regarding toxicity and selectivity. Many current medicines correlate to natural products, even in its early stages of development, which highlight the importance of natural sources in the design of new bioactive compounds. Capsaicin is the substance responsible for the pungency of Capsicum genus\' chili peppers. Capsaicin has been shown as an important natural agent because of its cytotoxic activity against several tumor cell lines. However, the pungency and poor stability presented by capsaicin restricts its therapeutic employments. Thus, the capsaicinoid scaffold has a great potential to inspire the design of new bioactive analog entities to be employed in anticancer therapy. This project aimed at designing and synthesizing sulfonamide and sulfonate analogues of capsaicin, performing molecular changes on capsaicinoid core. After a single step synthesis, the analogues were purified and characterized by melting point, 1H/13C NMR and elementary analysis. Subsequently, cytotoxicity assays were performed by MTT method on tumor cell lines MDA-MB-231 and MCF-7 (breast cancer), B16-F10 (murine melanoma) and on healthy fibroblast (3T3). Four compounds showed interesting biological activity at micromolar concentrations (&#181;M) and their IC50 were lower than that presented by capsaicin. The most active compounds, RPF-101 and RPF-151, had their death induction mechanisms investigated. In silico subsequent studies with the most active compounds (RPF-101 and RPF-151) showed that molecular changes promoted on each one increased their hydrophilicity and raised their dipole moment, which may be related to the improvement on biological activity of both. The evaluation of active compounds for Lipinski´s Rule of Five (RO5) properties revealed that they respect all properties presenting its druglikeness for oral administration since RPF151 did not presented in vivo pungency. The RPF-101 and RPF-151 indicate the success obtained by the structural optimization promoted on capsaicinoid scaffold and emphasizes the potential of these analogues to inspire the designing of new future optimized structures, with greater potency and less associated toxicity.

Foram desenvolvidos três experimentos para avaliação de uma mistura de extratos vegetais de orégano, pimenta e canela na produção de frangos de corte: Experimento I - Utilizou-se 320 pintos machos de um dia de idade da linhagem Cobb 500, distribuídos em delineamento experimental inteiramente casualisado com quatro tratamentos (0, 75, 150 e 225 ppm de extratos vegetais) com dez repetições de oito aves cada, criadas até 21 dias para determinação da digestibilidade dos nutrientes das dietas, pela metodologia de coleta do conteúdo ileal. Experimento II - Utilizou-se 600 pintos machos de um dia de idade da linhagem Cobb 500, distribuídos em delineamento experimental inteiramente casualisado com cinco tratamentos (Controle Positivo (CP) com 54 ppm de Bacitracina de Zinco; Controle Negativo (CN) sem aditivos melhoradores de desempenho; CN com a adição de 75 ppm; 150 ppm e 225 ppm de extratos vegetais) com dez repetições de doze aves cada para avaliação de desempenho e características de carcaça. Experimento III - Utilizou-se 600 pintos machos de um dia de idade da linhagem Cobb 500, distribuídos em delineamento experimental inteiramente casualisado com cinco tratamentos (Controle Positivo (CP) com 54 ppm de Bacitracina de Zinco; Controle Negativo (CN) sem aditivos melhoradores de desempenho; CP até 33 dias e posteriormente substituído pelo CN, no período de 34 a 42 dias de idade, com a adição de 75 ppm; 150 ppm e 225 ppm de extratos vegetais) com dez repetições de doze aves cada para avaliação de desempenho e características. Todos os dados foram analisados pelo Statistical Analysis System (SAS Institute Inc., 2008), submetidos à análise de regressão polinomial pelo procedimento GLM ao nível de 5% de significância. No experimento I, não foi observado efeito significativo entre os tratamentos para digestibilidade ileal da energia bruta, e a digestibilidade ileal da proteína bruta e dos aminoácidos apresentou efeito quadrático (P<0,05). No experimento II o consumo de ração não apresentou efeito significativo entre os tratamentos, já para ganho de peso e conversão alimentar foi observado efeito significativo (P<0,05) na comparação entre o antibiótico e os níveis de extratos vegetais adicionados a dieta. No experimento III, não foi observado efeito significativo entre os tratamentos para consumo de ração e ganho de peso, mas a conversão alimentar apresentou efeito significativo (P<0,05) na comparação entre o antibiótico e os níveis de extratos vegetais adicionados a dieta. Nos experimentos II e III, para as características de carcaça não foram observados efeitos significativos (P<0,05) entre os tratamentos. Concluiu-se que a inclusão de uma mistura de extratos vegetais de orégano, pimenta e canela na dieta de frangos de corte em substituição aos aditivos melhoradores de desempenho proporcionou maior digestibilidade ileal da proteína bruta e dos aminoácidos, e de acordo com os níveis praticados, os extratos vegetais não proporcionaram efeitos positivos no desempenho quando comparados com o tratamento com antibiótico, no período total de criação. Entretanto, o fornecimento na fase final apresentou um comportamento diferente, melhorando o rendimento de peito. / We developed three experiments to evaluate a mixture of herbal extracts of oregano, pepper and cinnamon in the production of broilers: Experiment I - was used 320 male chicks from one day old Cobb 500, distributed in completely randomized design with four treatments (0, 75, 150 and 225 ppm of plant extracts) with ten replicates of eight birds each, created up to 21 days to determine the nutrient digestibility of diets, the collection methodology of the ileal content. Experiment II - was used 600 male chicks from one day old Cobb 500, distributed in completely randomized design with five treatments (Positive Control (PC) with 54 ppm Bacitracin Zinc; Negative Control (NC) without enhancing additives performance; CN with the addition of 75 ppm, 150 ppm and 225 ppm of plant extracts) with ten replicates of twelve chicks each to assess performance and carcass characteristics. Experiment III - was used 600 male chicks from one day old Cobb 500, distributed in completely randomized design with five treatments (Positive Control (PC) with 54 ppm Bacitracin Zinc; Negative Control (NC) without enhancing additives performance; CP up to 33 days and subsequently replaced by CN, from 34 to 42 days of age, with the addition of 75 ppm, 150 ppm and 225 ppm plant extracts), with ten replicates of twelve chickens each for performance evaluation and characteristics. All data were analyzed using Statistical Analysis System (SAS Institute Inc., 2008), submitted to polynomial regression analysis by GLM procedure at the 5% level of significance. In the first experiment, no significant effect was observed between treatments for ileal digestibility of raw energy and ileal digestibility of crude protein and amino acids showed a quadratic effect (P <0.05). In experiment II feed intake had no significant effect between treatments, as for weight gain and feed conversion was no significant effect (P <0.05) in the comparison between the antibiotic and the levels of plant extracts added to the diet. In experiment III, no significant effect was observed between treatments for feed intake and weight gain, feed conversion but significant effect (P <0.05) in the comparison between the antibiotic and the levels of plant extracts added to the diet. In experiments II and III, for carcass traits were not significant effects (P <0.05) among treatments. It was concluded that the inclusion of a mixture of herbal extracts of oregano, pepper and cinnamon in the diet of broilers in place of performance enhancing additives provided higher ileal digestibility of crude protein and amino acids, and according to the levels prevailing, vegetable extracts yielded no positive effect on performance when compared with the antibiotic treatment, the total periods. However, the provision in the final phase showed a different behavior by improving the yield of breast.

Capsaicinoids are a class of compounds responsible for the "heat" of hot peppers. Capsaicin and dihydrocapsaicin have the highest burning effect. The aim of this work is to separate and quantify the two major capsaicinoids in fruits harvested at different stages of development and at different seasons. Simple and rapid HPLC method involves 73:27% methanol water mobile phase with C18 stationary phase and UV-Vis detector set at 210 nm. The method showed good reproducibility with 1.74% - 4.72% relative standard deviations, a linear response within 0.65–45.5 and 0.25-17.5 μg/mL for capsaicin and dihydrocapsaicin, respectively. The method achieved average recovery of 106% for capsaicin and 102% dihydrocapsaicin. Determination of capsaicinoids in four naturally grown chili and commercial source habanero were analyzed. The amount in the sample ranged from 1184-8156 μg/g for capsaicin and 430-3299 μg/g for dihydrocapsaicin.