O tabagismo é um problema mundial de saúde pública e é considerado a principal causa de morte evitável no mundo associado com câncer de pulmão, doença pulmonar obstrutiva crônica e infarto agudo do miocárdio. O tabagismo induz alterações morfológicas e funcionais no sistema respiratório. O transporte mucociliar (TMC) é um dos principais mecanismos de defesa do sistema respiratório que pode ser alterado com a fumaça e outros produtos do cigarro. O objetivo desse estudo foi avaliar os efeitos do tabagismo e da cessação do tabagismo no TMC nasal, nas propriedades do muco e sobre marcadores inflamatórios. Trinta e três indivíduos tabagistas foram incluídos no estudo após concordância com o termo de consentimento livre e esclarecido. O recrutamento de voluntários foi realizado na Faculdade de Medicina da Universidade de São Paulo (FMUSP) e no Ambulatório de Cessação do Tabagismo da Disciplina de Pneumologia do Hospital das Clínicas da FMUSP. As variáveis desfecho foram o TMC nasal analisado por meio do teste de trânsito da sacarina, as propriedades do muco por meio do ângulo de contato e da transportabilidade da tosse por alto fluxo e a quantificação de células inflamatórias e concentração de interleucinas (IL)-6 e IL-8 em lavado nasal. Vinte cessadores (idade média: 51 anos, 9 do sexo masculino) foram avaliados no tempo basal do estudo, 1o mês, 3o mês e 12o mês de cessação do tabagismo e 13 tabagistas (média de idade: 52 anos, 6 do sexo masculino) foram avaliados no tempo basal e 12 meses após o basal. As características demográficas, hábito tabágico inicial e morbidades de tabagistas e de cessadores foram similares. No tempo basal do estudo, os tabagistas e cessadores apresentaram disfunção do TMC nasal (17,9 ± 10,1 min e 17,4 ± 7,7 min, respectivamente, p=0,880). A cessação do tabagismo induziu melhora significativa do TMC nasal no 1o mês, 3º mês e 12o mês em 63%, 76% e 85% dos indivíduos, respectivamente. Somente aos 12 meses, foi possível observar melhora na transportabilidade do muco por alto fluxo (~ 23%), porém com aumento do número de macrófagos (2x) em lavado nasal. Não observamos alterações no ângulo de contato do muco e nas concentrações de citocinas em lavado nasal. Concluímos que a cessação do tabagismo induz melhora rápida no TMC nasal, porém melhora nas propriedades do muco foi observada somente após 12 meses de cessação do tabagismo / Smoking is a health problem in the world. It is considered a main cause of preventable death and is associated with lung cancer, chronic obstructive pulmonary disease and myocardium infarction. Smoking induces morphological and functional changes in the respiratory system. Mucociliary clearance (MCC) is one of the main defense mechanisms of the respiratory system that can be affected by smoke and other cigarette products. The aim of this study was to assess the effects of smoking and smoking cessation on nasal MCC, mucus properties and inflammatory biomarkers. Thirty three smokers were included in this study after agreement with the written informed consent. Subject´s recruitment was performed at Faculdade de Medicina da Universidade de São Paulo (FMUSP) and Ambulatório de Cessação do Tabagismo da Disciplina de Pneumologia do Hospital das Clínicas da FMUSP. The outcome variables were nasal MCC evaluated by saccharin transit test, mucus properties using contact angle and mucus transportability by high airflow and quantification of inflammatory cells number and interleukin (IL)-6 and IL-8 in the nasal lavage fluid. Twenty volunteers in the smoking cessation program (mean age: 51 years, 9 male) were assessed at baseline, month 1, month 3 and month 12 after of the smoking cessation and 13 smokers (mean age: 52 years, 6 male) were assessed at baseline and 12 months after baseline. Demographic characteristics, smoking history and morbidities were similar between the two groups. At baseline, smokers showed impaired nasal MCC (17.9 ± 10.1 min and 17.4 ± 7.7 min, respectively, p=0.880). Smoking cessation significantly improved nasal MCC at 1 month, 3 months and 12 months in 63%, 76% and 85% of the subjects, respectively. Only after 12 months of smoking cessation, improvement in mucus transportability by high airflow (~ 23%) was observed, however, with increased number of macrophages (2-fold) in nasal lavage fluid. No changes were observed in mucus contact angle and cytokines concentrations in nasal lavage fluid. We concluded that smoking cessation induces rapid improvement in nasal MCC, however, improvement in mucus properties were observed only after 12 months of smoking cessation

Introdução: Uma imunossupressão eficaz é fundamental para a sobrevivência do paciente após o transplante de pulmão. Estudos prévios demonstraram que drogas imunossupressoras como ciclosporina A, tacrolimo, micofenolato de sódio e prednisona prejudicaram a depuração das vias aéreas de ratos. Para prevenir a rejeição aguda, o basiliximab tem sido utilizado como terapia de indução previamente ao transplante de pulmão em muitos centros ao redor do mundo. Entretanto, existem poucos trabalhos reportando seus efeitos adversos. Objetivo: Avaliar se o basiliximab isolado e em conjunto com a terapia tríplice (tacrolimo, micofenolato de sódio e prednisona) causa efeitos adversos no aparelho mucociliar traqueobrônquico, alterando a depuração mucociliar das vias aéreas de ratos. Método: Oitenta ratos foram distribuídos em quatro grupos conforme o tratamento: Controle, Basiliximab, Tríplice e Basiliximab+Tríplice; e dois subgrupos de acordo com o tempo de tratamento: 7 e 15 dias. Após o período de tratamento, os animais foram eutanasiados e as seguintes análises foram realizadas: análise do lavado broncoalveolar, frequência de batimento ciliar (FBC), velocidade de transporte mucociliar (VTMC), transportabilidade do muco in vitro, histologia, expressão do gene Muc5ac, concentração da proteína mucina do gene Muc5ac e avaliação de apoptose celular no epitélio das vias aéreas. Resultados: Não houve alteração do número de leucócitos no lavado broncoalveolar e da FBC entre os grupos. Já a VTMC foi menor nos grupos Basiliximab e Basiliximab+Tríplice tratados por 7 dias, enquanto nos animais tratados por 15 dias a velocidade foi menor nos grupos Trípice e Basiliximab+Tríplice. O transporte do muco in vitro foi menor no grupo Basiliximab+Tríplice tratado por 15 dias. A porcentagem dos mucos ácido e neutro não foi diferente entre os grupos tratados por 7 e 15 dias, o mesmo ocorreu para a expressão e concentração da proteína mucina do gene Muc5ac. Os grupos Tríplice e Basiliximab+Tríplice tratados por 7 e 15 dias, respectivamente, apresentaram número maior de células apoptóticas no epitélio da via aérea. Conclusão: A droga basiliximab, isolada e em conjunto com a terapia tríplice, prejudicou o aparelho mucociliar traqueobrônquico de ratos, especificamente a depuração mucociliar / Introduction: Optimal immunosuppression is critical to the survival of the patient after lung transplantation. Previous studies showed that immunosuppressive drugs such as cyclosporine, tacrolimus, sodium mycophenolate and prednisone impaired mucociliary clearance of rats. To prevent acute rejection, basiliximab has been used as induction therapy before lung transplantation in many centers around the world. However, there are few studies reporting its side effects. Objective: Evaluate if basiliximab alone and in combination with triple therapy (tacrolimus, sodium mycophenolate and prednisone) causes adverse effects on the tracheobronchial mucociliary apparatus by impairing airways mucociliary clearance of rats. Method: Eighty rats were divided into four groups according to treatment: Control, Basiliximab, Triple and Basiliximab+Triple; and according to the treatment time: 7 and 15 days. After the treatment period, the animals were euthanized and the following analyzes were performed: bronchoalveolar lavage, ciliary beating frequency (CBF), mucociliary transport velocity (MCTV), mucus transportability rate in vitro, hystology, Muc5ac gene expression, concentration of the mucin protein of the Muc5ac gene and evaluation of cellular apoptosis in the airway epithelium. Results: There was no alteration in the number of leukocytes in the bronchoalveolar lavage fluid and the CBF between groups. The MCTV was lower in the Basiliximab and Basiliximab+Triple groups treated for 7 days, while the velocity was lower in the Triple and Basiliximab+Triple groups treated for 15 days. The mucus transportability rate in vitro was lower in the Basiliximab+Triple group treated for 15 days. There was no difference in percentage of both acidic mucus and neutral mucus between groups treated for 7 and 15 days. Also, there was no difference in the expression and concentration of the mucin of the Muc5ac gene. The Triple and Basiliximab+Triple groups treated for 7 and 15 days, respectively, had a higher number of apoptotic cells in the airway epithelium. Conclusion: The basiliximab, alone and in conjunction with triple therapy, impaired the tracheobronchial mucociliary apparatus of rats, specifically the mucociliary clearance

Les mucines sont les molécules structurales de base du gel de mucus qui assure la protection et la lubrification de la muqueuse gastro-intestinale. Notre but était de déterminer l'effet de la leptine sur la sécrétion et l'expression des mucines gastro-intestinales. Cette étude a été réalisée in vivo chez le rat et in vitro à l'aide des lignées cellulaires de rat DHE et humaine HT29-MTX. Nous avons trouvé que ces 2 lignées expriment les récepteurs (Ob-R) de la leptine. La perfusion luminale de la leptine dans le côlon de rat ainsi que la stimulation des cellules DHE par la leptine a entraîné une augmentation de la sécrétion de mucines et du niveau des ARNm codant pour rMuc2, rMuc3 et rMuc4. Cet effet de la leptine était dose dépendant. Testée dans les HT29-MTX, la leptine a provoqué une augmentation de la sécrétion de mucines ainsi qu'une augmentation des transcrits de MUC2, MUC4 et MUC5AC via la voie de la PKC, la PI3K et les MAPK. Ces résultats montrent que la leptine pourrait jouer un rôle essentiel dans la protection de la muqueuse colique.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The commercial formulations of glyphosate herbicide have been widely used in agriculture and pisciculture for controlling plant weeds. Fish can be affected when drainage water reaches water courses, causing a disturbance in the aquatic ecosystem. Therefore, the aim of this study was to investigate whether exposure to a commercial formulation of glyphosate affects toxicological parameters in piavas (Leporinus obtusidens) and jundiás (Rhamdia quelen). Piavas were exposed to: 0 (control), 3, 6, 10 or 20 mg/L of glyphosate (480 g/L) and jundiás to 0 (control), 0.2 or 0.4 mg/L, both for 96 h. Enzymatic (AChE), hematological (hematocrit, hemoglobin and erythrocyte and leucocyte count) and metabolic (glucose, glycogen, lactate, protein and ammonia) parameters were analyzed in different tissues of these species. In addition, oxidative stress parameters, such as activity of the antioxidant enzyme catalase, protein carbonilation and TBARS levels were analyzed. The results showed that brain acetylcholinesterase (AChE) activity decreased significantly in both species exposed, but there was no significant change in the muscle tissue. There was a decrease in hematological parameters in blood of piavas. After exposure to this herbicide, both species demonstrated metabolic disorders. In piavas, there was a reduction of glycogen and an increase of glucose in hepatic tissue, and a reduction of muscle glycogen and glucose. Lactate and protein showed a decrease in hepatic tissue after exposure to all concentrations of the herbicide, but in muscle tissue there was an increase in these metabolites. The levels of ammonia increased in both tissues after exposure, varying with the glyphosate concentration. In jundiás, there was an increase of hepatic glycogen, but a reduction of muscle glycogen at both concentrations tested. Glucose showed a decrease in liver and increase in muscle. After exposure to this herbicide, the levels of lactate were increased in both tissues. The levels of protein were increased in hepatic tissue and decreased in muscle tissue, while the levels of ammonia showed an increase at both herbicide concentrations in both tissues tested. Catalase activity showed an increase in jundiás, but not in piavas. TBARS levels showed an elevation in muscle tissue in jundiás while in piavas there was an increase in hepatic tissue. An increase in protein carbonyl formation in liver of jundiás was observed. In addition, there was an increase in mucus parameters in piavas. These results indicate that the parameters measured may be good indicators of herbicide contamination in piavas and jundiás of the southern region of Brazil. / As formulações comerciais do herbicida glyphosate têm sido amplamente utilizadas na agricultura e na piscicultura para controle de plantas daninhas. Os peixes podem ser afetados quando as águas de drenagem atingem os cursos d água, acarretando um desequilíbrio no ecossistema aquático. Sendo assim, o objetivo deste estudo foi avaliar o efeito de uma formulação comercial de glyphosate sobre parâmetros toxicológicos em piavas (Leporinus obtusidens) e jundiás (Rhamdia quelen). Piavas foram expostas à: 0 (controle), 3, 6, 10 ou 20 mg/L de glyphosate (480 g/L) e os jundiás à 0 (controle), 0,2 ou 0,4 mg/L , ambos por 96 h. Os parâmetros analisados foram enzimáticos (AChE), hematológicos (hematócrito, hemoglobina, contagem de eritrócitos e de leucócitos) e metabólicos (glicose, glicogênio, lactato, proteína e amônia) em diferentes tecidos destas espécies. Ademais, analisaram-se parâmetros de estresse oxidativo, como a atividade da enzima antioxidante catalase, carbonilação de proteínas e níveis de TBARS. Os resultados mostraram que a atividade da acetilcolinesterase (AChE) cerebral diminuiu significativamente em ambas as espécies expostas, porém no tecido muscular não se observaram alterações significativas. Houve um decréscimo nos parâmetros hematológicos no sangue de piavas expostas. Após exposição ao glyphosate, as duas espécies demonstraram desordens metabólicas. Em piavas, ocorreu uma redução nos níveis de glicogênio e um aumento nos níveis de glicose no tecido hepático, porém uma significante redução do glicogênio e glicose muscular. As concentrações de lactato e proteína apresentaram uma diminuição no tecido hepático após exposição a todas as concentrações deste herbicida, porém no tecido muscular houve um aumento destes metabólitos. Os níveis de amônia aumentaram em ambos tecidos após exposição às diferentes concentrações do glyphosate. Em jundiás, a glicose esteve reduzida no fígado e aumentada no músculo. Houve um aumento do glicogênio hepático, mas uma redução do glicogênio muscular em ambas as concentrações testadas. Após exposição a este herbicida, os níveis de lactato estavam aumentados em ambos tecidos. Os níveis de proteína estavam aumentados no tecido hepático e diminuídos no tecido muscular, enquanto os níveis de amônia estavam aumentados em ambas as concentrações e tecidos testados. A atividade da enzima antioxidante catalase apresentou um aumento em jundiás expostos, porém em piavas não houve alterações. Os níveis de TBARS em jundiás mostraram uma elevação no tecido muscular enquanto em piavas houve um aumento no tecido hepático. Observou-se um aumento na formação de proteína carbonila em fígado de jundiás. Além disso, houve aumento nos parâmetros do muco (glicose e proteína) em piavas expostas. Estes resultados indicam que os parâmetros medidos podem ser bons indicadores da contaminação deste herbicida em piavas e jundiás da região Sul.

Introdução: Uma imunossupressão eficaz é fundamental para a sobrevivência do paciente após o transplante de pulmão. Estudos prévios demonstraram que drogas imunossupressoras como ciclosporina A, tacrolimo, micofenolato de sódio e prednisona prejudicaram a depuração das vias aéreas de ratos. Para prevenir a rejeição aguda, o basiliximab tem sido utilizado como terapia de indução previamente ao transplante de pulmão em muitos centros ao redor do mundo. Entretanto, existem poucos trabalhos reportando seus efeitos adversos. Objetivo: Avaliar se o basiliximab isolado e em conjunto com a terapia tríplice (tacrolimo, micofenolato de sódio e prednisona) causa efeitos adversos no aparelho mucociliar traqueobrônquico, alterando a depuração mucociliar das vias aéreas de ratos. Método: Oitenta ratos foram distribuídos em quatro grupos conforme o tratamento: Controle, Basiliximab, Tríplice e Basiliximab+Tríplice; e dois subgrupos de acordo com o tempo de tratamento: 7 e 15 dias. Após o período de tratamento, os animais foram eutanasiados e as seguintes análises foram realizadas: análise do lavado broncoalveolar, frequência de batimento ciliar (FBC), velocidade de transporte mucociliar (VTMC), transportabilidade do muco in vitro, histologia, expressão do gene Muc5ac, concentração da proteína mucina do gene Muc5ac e avaliação de apoptose celular no epitélio das vias aéreas. Resultados: Não houve alteração do número de leucócitos no lavado broncoalveolar e da FBC entre os grupos. Já a VTMC foi menor nos grupos Basiliximab e Basiliximab+Tríplice tratados por 7 dias, enquanto nos animais tratados por 15 dias a velocidade foi menor nos grupos Trípice e Basiliximab+Tríplice. O transporte do muco in vitro foi menor no grupo Basiliximab+Tríplice tratado por 15 dias. A porcentagem dos mucos ácido e neutro não foi diferente entre os grupos tratados por 7 e 15 dias, o mesmo ocorreu para a expressão e concentração da proteína mucina do gene Muc5ac. Os grupos Tríplice e Basiliximab+Tríplice tratados por 7 e 15 dias, respectivamente, apresentaram número maior de células apoptóticas no epitélio da via aérea. Conclusão: A droga basiliximab, isolada e em conjunto com a terapia tríplice, prejudicou o aparelho mucociliar traqueobrônquico de ratos, especificamente a depuração mucociliar / Introduction: Optimal immunosuppression is critical to the survival of the patient after lung transplantation. Previous studies showed that immunosuppressive drugs such as cyclosporine, tacrolimus, sodium mycophenolate and prednisone impaired mucociliary clearance of rats. To prevent acute rejection, basiliximab has been used as induction therapy before lung transplantation in many centers around the world. However, there are few studies reporting its side effects. Objective: Evaluate if basiliximab alone and in combination with triple therapy (tacrolimus, sodium mycophenolate and prednisone) causes adverse effects on the tracheobronchial mucociliary apparatus by impairing airways mucociliary clearance of rats. Method: Eighty rats were divided into four groups according to treatment: Control, Basiliximab, Triple and Basiliximab+Triple; and according to the treatment time: 7 and 15 days. After the treatment period, the animals were euthanized and the following analyzes were performed: bronchoalveolar lavage, ciliary beating frequency (CBF), mucociliary transport velocity (MCTV), mucus transportability rate in vitro, hystology, Muc5ac gene expression, concentration of the mucin protein of the Muc5ac gene and evaluation of cellular apoptosis in the airway epithelium. Results: There was no alteration in the number of leukocytes in the bronchoalveolar lavage fluid and the CBF between groups. The MCTV was lower in the Basiliximab and Basiliximab+Triple groups treated for 7 days, while the velocity was lower in the Triple and Basiliximab+Triple groups treated for 15 days. The mucus transportability rate in vitro was lower in the Basiliximab+Triple group treated for 15 days. There was no difference in percentage of both acidic mucus and neutral mucus between groups treated for 7 and 15 days. Also, there was no difference in the expression and concentration of the mucin of the Muc5ac gene. The Triple and Basiliximab+Triple groups treated for 7 and 15 days, respectively, had a higher number of apoptotic cells in the airway epithelium. Conclusion: The basiliximab, alone and in conjunction with triple therapy, impaired the tracheobronchial mucociliary apparatus of rats, specifically the mucociliary clearance

Les voies aériennes humaines sont bordées d'un épithélium pseudostratifié composé principalement de cellules basales et de cellules pyramidales parmi lesquelles figurent les cellules sécrétrices de mucus et les cellules multiciliées. Toutes ces cellules contribuent à la clairance mucociliaire des voies respiratoires. Cet épithélium se régénère lentement dans des conditions homéostatiques, mais il est capable de se régénérer rapidement après agression grâce à des processus de prolifération, de migration, de polarisation et de différenciation. Chez les patients atteints de maladies respiratoires chroniques telles que la broncho-pneumopathie chronique obstructive, l'asthme ou la mucoviscidose, la réparation tissulaire est souvent défectueuse, caractérisée par une perte de cellules multiciliées et une hyperplasie des cellules sécrétrices, ayant pour conséquence une clairance mucociliaire affectée. La séquence des événements cellulaires conduisant à un tissu fonctionnel ou remodelé est encore mal décrite. Notre principal objectif a été d’identifier les types cellulaires successifs mis en jeu lors de la régénération tissulaire et les événements moléculaires responsables d'une régénération saine ou pathologique. Nous avons analysé la composition cellulaire de l’épithélium des voies respiratoires à plusieurs stades de différenciation en utilisant un modèle de culture 3D in vitro qui reproduit la composition cellulaire in vivo. En appliquant une méthode de transcriptomique sur cellule unique couplée à des méthodes bioinformatiques, nous avons établi les hiérarchies cellulaires permettant de reconstruire les différentes trajectoires cellulaires mises en jeu lors de la régénération de l’épithélium des voies respiratoires humaines. Après avoir confirmé les lignages cellulaires qui ont été précédemment décrits, nous avons découvert une nouvelle trajectoire reliant les cellules sécrétrices de mucus aux cellules multiciliées. Nous avons également caractérisé de nouvelles populations cellulaires et de nouveaux acteurs moléculaires impliqués dans le processus de régénération de l'épithélium des voies respiratoires humaines. Enfin, grâce à ces approches, nous avons mis en évidence des réponses spécifiques de chaque type cellulaire survenant dans des situations pathologiques d’hyperplasie sécrétoire. Ainsi, nos données, en apportant d'importantes contributions à la compréhension de la dynamique de différenciation de l’épithélium des voies respiratoires humaines, ouvrent de nouvelles voies vers l’identification de cibles thérapeutiques. / Human airways are lined by a pseudostratified epithelium mainly composed of basal and columnar cells, among these cells we can find multiciliated, secretory cells and goblet cells. All these cells work together in the mucociliary clearance of the airways. This epithelium regenerates slowly under homeostatic conditions but is able to recover quickly after aggressions through proliferation, migration, polarization and differentiation processes. However, in patients with chronic pulmonary diseases such as chronic obstructive pulmonary disease, asthma or cystic fibrosis, epithelial repair is defective, tissue remodeling occurs, leading to loss of multiciliated cells and goblet cell hyperplasia, impairing correct mucociliary clearance. The sequence of cellular events leading to a functional or remodelled tissue are still poorly described. Hence, we aim at identifying the successive cell types appearing during tissue regeneration and the molecular events that are responsible for healthy or pathological regeneration. We have analysed airway epithelial cell composition at several stages of differentiation using an in vitro 3D culture model which reproduces in vivo epithelial cell composition. Applying single cell transcriptomics and computational methods, we have identified cell lineage hierarchies and thus constructed a comprehensive cell trajectory roadmap in human airways. We have confirmed the cell lineages that have been previously described and have discovered a novel trajectory linking goblet cells to multiciliated cells. We have also discovered novel cell populations and molecular interactors involved in the process of healthy human airway epithelium regeneration. Using these approaches, we have finally shed light on cell-type specific responses involved in pathological goblet cell hyperplasia. Our data, by bringing significant contributions to the understanding of differentiation’s dynamics in the context of healthy and pathological human airway epithelium, may lead to the identification of novel therapeutic targets.

El concepto de alimento funcional se emplea para describir a aquellos alimentos adicionados con ingredientes de diversas clases y orígenes que pueden ejercer efectos beneficiosos sobre quien los ingiere. Este concepto surgido en Japón ha ido popularizándose y expandiéndose hacia otros continentes como Europa fundamentalmente debido a la preocupación sobre la nutrición, la dieta y la salud de la sociedad actual. Los probióticos representan una gran área dentro de los alimentos funcionales y se ha intensificado la investigación para desarrollar productos probióticos y profundizar en el conocimiento de los efectos sobre la salud humana. Los probióticos se definen como "suplementos alimentarios microbianos vivos que afectan de forma ventajosa al animal huésped mejorando su equilibrio intestinal microbiano". Son microorganismos que estimulan las funciones protectoras del tracto digestivo, y también son conocidos como bioterapéuticos, bioprotectores o bioprofilácticos, ya que se utilizan para prevenir las infecciones entéricas y gastrointestinales. Un microorganismo probiótico efectivo debe poseer una serie de características: no ha de ser no patógeno ni tóxico, debe ejercer efectos beneficiosos sobre la salud de quien lo ingiere, tener origen humano, ha de ser tecnológicamente utilizable, ha de presentar un elevado porcentaje de células viables, debe ser capaz de sobrevivir a la flora intestinal, ha de permanecer viable durante su almacenamiento en refrigeración, y tener capacidad de adherirse a la superficie mucosa, etc. El establecimiento de criterios de selección y controles de calidad para productos probióticos se considera una prioridad debido a la rápida incorporación de estos productos en el mercado y su distribución en el ámbito internacional sin la existencia previa de una normativa comúnmente aceptada. En los últimos años ha aumentado el interés por los productos elaborados con microorganismos probióticos, pertenecientes a los géneros Lactobacillus y Bifidobac / Collado Amores, MC. (2005). Caracterización de cepas del género Bifidobacterium con carácter probiótico [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/1907 / Palancia

MPH / Department of Public Health / Despite the availability of the South African cervical cancer screening guidelines at clinics, women are still seen in the out-patient department of selected rural district hospital in Thulamela Municipality, referred from Primary Health Care (PHC) facilities with no cervical cancer screening results. Thus, cervical cancer screenings are done at a hospital and results often came back positive for cervical cancer. Such practices pose a risk of delayed cervical cancer diagnoses and its discovery at an advanced stage, increasing cervical cancer mortality rates. The purpose of this study was to investigate the compliance of professional nurses at primary health care facilities to the South African cervical cancer screening guidelines in Thulamela Municipality. This study adopted a qualitative approach as it aimed to get a better understanding on compliance of professional nurses regarding South Africa’s cervical cancer screening guidelines. Nonprobability sampling was used to select primary health care facilities including interested participants who were judged to possess the information that was required to answer the research question of this study. The sample size of the study was determined by data saturation. Trustworthiness was ensured by four criteria of Guba and Linclon, namely credibility, transferability, dependability and confirmability. A digital recorder was used to log individual responses during interview sessions. Data from the digital recordings were transcribed verbatim. Results were analysed and interpreted thematically. This study established that clinic professional nurses are non –compliance to the South African cervical cancer screening guidelines; nurses face several challenges such as inadequate knowledge of the cervical cancer screening guidelines, shortage of resources and shortage of staff. Recommendations that emanated from the discussion of the findings and the conclusion of this study are likely to have implications and applications for supporting and advancing the cervical cancer screenings. / NRF

INTRODUÇÃO: A toxina botulínica tipo A (TXB-A) tem sido testada no tratamento da rinite, principalmente nos casos de rinite idiopática. Sugere-se que um estado de hiper-reatividade do nervo trigêmeo esteja envolvido na fisiopatologia da rinite idiopática. O nervo trigêmeo possui fibras sensitivas não mielinizadas tipo C (FSNMT-C) que contém os neuropeptídeos substância P (SP) e peptídeo relacionado ao gene da calcitonina (CGRP). O óxido nítrico (NO) produzido pelas enzimas óxido nítrico sintase (NOS) também está envolvido nesse processo de neurorregulação nasal. O transporte mucociliar, mecanismo primário de defesa do sistema respiratório, é formado pelo batimento ciliar e muco nasal, e esses componentes podem ser influenciados por diferentes neuropeptídeos e neurotransmissores presentes na mucosa nasal. OBJETIVO: O objetivo deste estudo foi avaliar o efeito da TXB-A sobre a expressão da SP, CGRP e óxido nítrico sintase neural (nNOS), além de sua influência sobre o transporte mucociliar nasal em coelhos. MÉTODOS: Coelhos machos saudáveis da linhagem Nova Zelândia foram divididos em dois grupos: o grupo tratamento recebeu TXB-A (25UI) na concha nasomaxilar (CNM) do lado direito e soro fisiológico a 0,9% (SF0,9%) na CNM esquerda. O grupo controle recebeu SF0,9% na CNM direita e nenhuma intervenção na CNM esquerda. Foram investigados os efeitos da TXB-A sobre a expressão da SP, CGRP e nNOS no tecido de CNM por meio da imuno-histoquímica. Para esta análise, dividiu-se o tecido em camada externa (CE, acima da membrana basal) e camada interna (CI, abaixo da membrana basal). Avaliou-se também a presença de apoptose celular, a frequência de batimento ciliar (FBC), o perfil histoquímico do muco nasal (glicoproteínas ácidas e neutras) e a espessura do epitélio (ESP-CE). RESULTADOS: Foi observado um aumento significativo na quantidade de células apoptóticas na CNM do grupo tratamento que recebeu TXB-A em comparação aos controles (p <= 0,001). A CNM do grupo tratamento que recebeu SF0,9% exibiu um aumento na quantidade de células apoptóticas na CI ao comparar com os controles (CNM SF0,9%, p=0,035) (CNM sem intervenção, p=0,022), e também um aumento da expressão de SP na CE em comparação aos controles (CNM SF0,9%, p=0,021) (CNM sem intervenção, p=0,040). A expressão de CGRP apresentou um aumento na CNM do grupo tratamento que recebeu TXB-A em comparação à CNM sem intervenção (p=0,008). A FBC, expressão de nNOS, perfil histoquímico do muco nasal e ESP-CE não apresentaram diferenças significativas. DISCUSSÃO: O aumento da expressão de CGRP e SP pode ter sido causado por inibição de sua exocitose vesicular pela TXB-A, levando ao seu acúmulo intracelular. Não foram observadas diferenças significativas na FBC ou perfil histoquímico do muco nasal, indicando que o aumento da expressão de CGRP e SP não foi associado à inflamação. O aumento da quantidade de células apoptóticas e da expressão de SP na CNM SF0,9% do grupo tratamento pode ter sido causado por um efeito central da TXB-A. CONCLUSÃO: A administração nasal de TXB-A aumentou a expressão de CGRP e SP, possivelmente por acúmulo intracelular por causa da inibição da sua exocitose, mas sem alterar a FBC e o perfil histoquímico do muco nasal / INTRODUCTION: Botulinum toxin type A (BoNT-A) has been assessed in the treatment of rhinitis, especially in cases of idiopathic rhinitis. Trigeminal hyper-responsiveness appears to be involved in the pathological process of idiopathic rhinitis. Trigeminal nociceptive type C unmyelinated sensory fibers contain the neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP). Nitric oxide (NO) produced by the enzyme nitric oxide synthase (NOS) are also involved in this nasal neurorregulation process. The mucociliary clearance, primary defense system of the respiratory system, is composed by the ciliary beat and nasal mucus. These components can be influenced by different nasal neuropeptides and neurotransmitters. OBJECTIVE: The aim of this study was to evaluate the effect of BoNT-A on the expression of SP, CGRP and neural nitric oxide synthase (nNOS), and its influence on nasal mucociliary clearance in rabbits. METHODS: Healthy New Zealand male rabbits were divided into two groups: the treatment group was challenged with BoNT-A (25UI) in the right nasomaxillary turbinate (NMT) and saline (SF0.9%) in the left NMT. The control group received SF0.9% in the right NMT and no-intervention in the left NMT. We investigated the effects of BoNT-A on SP, CGRP and nNOS expression in the NMT tissue by immunohistochemistry. Each area of interest was subdivided into an internal layer (IL: below the basement membrane) and outer layer (OL: above the basement membrane) for analysis. It was also assessed signs of cellular apoptosis, ciliary beat frequency (CBF), mucus histochemical profile (acidic and neutral glycoproteins) and epithelial thickness (EP-TH). RESULTS: It was observed a significant increase in the amount of apoptotic cells in the BoNT-A-challanged NMT compared with controls (p <= 0.001). The NMT of treatment group which received only SF0.9% showed an increase in the amount of apoptotic cells in the IL compared with controls (NMT SF0.9%, p = 0.035) (NMT no-intervention, p = 0.022), and also an increase in the SP expression in the OL compared with controls (NMT SF0.9%, p = 0.021) (NMT no-intervention, p = 0.040). CGRP expression showed higher expression in the BoNT-A-challanged NMT compared with no-intervention NMT (p=0.008). The CBF, nNOS expression, mucus histochemical profile and EP-TH did not show significant differences. DISCUSSION: The increased CGRP and SP expression could be due to inhibition of vesicular exocytosis by BoNT-A, leading to CGRP and SP intracellular accumulation. No significant differences in CBF or mucus histochemical profile were observed, indicating that the increased CGRP and SP expression was not associated with inflammation. The increase in the amount of apoptotic cells and SP expression in the SF0.9% NMT of treatment group may be due to a central effect of BoNT-A. CONCLUSION: Nasal administration of BoNT-A increased SP and CGRP expression, possibly via inhibition of their release, but did not change the CBF or mucus profile

Ultraviolet light is damaging but fish have evolved protective mechanisms, which allows them to live in shallow water reefs, high in UV radiation. This thesis details my investigation into the physiological ecology of solar ultraviolet (UV) absorbing compounds, known as mycosporine-like amino acids found in the external epithelial mucus, and examines the supporting role potentially played by a UV-induced DNA repair mechanism in coral reef fish of the Indo-Pacific. Using reverse phase chromatography and UV spectrophotometry, I examined whether the distribution of MAA compounds across different areas of the body is correlated with differential UV exposure. Comparisons were made between the MAA content and the absorbance spectra of mucus from the dorsal, ventral, caudal and head body surface areas in five species of Scaridae (Chlorurus sordidus, Scarus schlegeli, S. niger, S. psittacus and S. globiceps) from Ningaloo Reef, Coral Bay, Western Australia. All fish analysed had at least five MAAs present, and results showed that fish had increased UV absorbance in mucus over the dorsal area, which receives the brunt of UV radiation. Little UV protection was found in mucus from the ventral area, which receives the lower level of UV radiation mostly via reflection of the sand and reef surfaces. Furthermore, UV absorbance per mg dry mucus versus standard fish length showed that there is a positive relationship in C. sordidus with increasing size. I examined whether there is a difference in the quantity of UV screening compounds found in the mucus of fish along a longitudinal geographical gradient from inshore reefs (Lizard Island, Great Barrier Reef) to the outer edge reefs to oceanic reefs (Osprey Reef). MAA absorbance increased with longitudinal distance from the mainland landmass of Australia to more oligotrophic outer reefs, where UV attenuation is reduced and the ocean is more transparent to UV wavelength. I determined that fish living on inshore, more turbid reefs where UV attenuation in shallow waters is high have lower levels of MAA protection than fish from clear oceanic reefs. Furthermore, there seems to be a direct relationship between light attenuation and exposure with the quantity of protective sunscreening found in the mucus of reef fish. It is know that UV irradiation decreases with water depth and that mucus from fish with deep habitats absorbs less UV than that of fish from shallow habitats. It is unknown however, whether this UV protection is variable within the same individuals and if so, how fast changes 11 occur. To test this, I relocated 9 ambon damselfish from a deep reef (18 m) to a shallow reef (1.5 m) to expose fish to increased levels of UV and relocated another 7 fish from a shallow to a deep reef to expose fish to decreased levels of UV. One week after relocation, all fish were returned to their original reef site to determine whether MAA levels would return to their initial levels. Fish relocated to a shallower depth were recovered and had a 60% (SD+/-2%) increase in mucus UV absorbance. Conversely, the fish relocated to a deeper depth were recovered and had a 41% (SD+/-1%) decrease mucus UV absorbance. No difference was found between UV absorbance of relocated and original fish at both depth. Six days after fish were returned to their original reef, mucus UV absorbance levels had returned to 67% +/- 4% of the original level. These results show that mucus UV absorbance is variable in individual ambon damselfish and that the sunscreen protection typical for a certain depth is reached in relocated fish within just a few days of relocation. The rate of MAA loss is higher than the accumulation of MAAs suggesting that diet is not the sole determining factor involved in the sequestration of MAAs to mucus. The cleaner fish Labroides dimidiatus performs a mutualistic service by removing ectoparasites such as gnathiid isopods as well other dead infected tissue from its clients. Cleaner fish however are also known to feed on client mucus. The benefits of eating mucus until recently were unclear. In this study, we analysed the mucus of several cleaner fish clients to determine whether mucus feeding has a nutritional advantage over gnathiids and whether cleaner fish obtain their own MAA protection through this dietary mucus ingestion. Results show that host fish that are infected with gnathiids of poor nutritional value, in contrast to those that harbour gnathiids with higher nutritional value, continuously exude mucus that has both high nutritional value and high MAA content. These findings support the conclusion that in a competitive market for cleaners some host fish are forced to offer more than parasites to cleaners. Ultraviolet light that is not filtered by UV absorbing compounds such as MAA may still lead to DNA damage such as the formation of cyclobutane pyrimidine dimers (CPDs) or 6-4 photoproducts (6-4 PPs). However, coral reef fish have alternative mechanisms to overcome UV induced damage via the photolyase DNA repair mechanisms. We experimentally demonstrated for the first time that a coral reef fish species, the moon wrasse Thalassoma lunare has the ability to repair DNA damage via photoreactivation. Fish both with and without MAA protection were irradiated with UVB wavelength to induce DNA lesions. Half of the experimental fish were then exposed to photoreactivating wavelength to induce DNA repair 12 while the other fish were blocked from the repair mechanisms. Fish which had undergone DNA repair had the lowest number of lesions regardless of mucus MAA protection. When fish were blocked from photoreactivation wavelengths MAA sunscreens clearly served a photoprotective role. The amount of damage was greatest in fish which both lacked MAAs and which were also blocked from photoreactivating wavelengths. Thus for the overall UV protection of fish both the MAA sunscreens as well as the DNA repair system play a significant role in counteracting UV damage. Ultraviolet protection by MAA sunscreens is ubiquitous in marine fish. To date the same 5 MAA compounds (palythine (λmax 320 nm), asterina (λmax 330 nm), palythinol (λmax 332 nm), usujirene (λmax 357 nm) and palythene (λmax 360nm) have been identified in the mucus of several different species of reef fish from Australia. Here we report the first evidence of the presence of additional UV absorbing compounds found in the mucus of fish from Indonesia. Using UV spectroscopy the mucus of four species of fish was compared between both geographical regions. The presence of an additional peak between 294-296 nm wavelengths suggests the presence of gadusol and/or deoxygadusol, which are photoprotective compounds, thought to be the precursors of MAAs. Thus, UV protecting compounds in the mucus of fish may not be as conserved between different regions as previously assumed. Our knowledge concerning the effect of UV radiation has advanced considerably in the past decade and my research findings contribute to the better understanding of protective mechanisms of marine fish. The correlations I have found between UV attenuation/exposure, depth, and longitude of sampled individuals lead me to believe that mucus UV absorbing MAA compounds are a highly efficient adaptive defence.