Desarrollo de nuevas formulaciones tópicas de anfotericina B para el tratamiento de micosis

Las infecciones por hongos se han convertido en un tema de gran preocupación en todo el mundo, se estima que más de 40 millones de personas sufren infecciones por hongos, tanto en países desarrollados como en países en vías de desarrollo (Güngör, et al., 2013). Las micosis superficiales se encuentran entre las formas más frecuentes de infecciones en los humanos. Se estima que afectan un 20-25 % de la población mundial y su incidencia está constantemente en aumento (Vena, et al., 2012) (Havlickova, et al., 2008) (Das, et al., 2007). Actualmente, este tipo de infecciones son un frecuente motivo de consulta para el médico de familia (Hernández, et al., 2014) y el dermatólogo. Lo cual nos obliga a permanecer constantemente actualizados La candidiasis es la micosis emergente con mayor efecto en el ser humano debido a su frecuencia y a la gravedad de sus complicaciones (López-Martínez, R., 2010). La candidiasis superficial es una de las formas clínicas más comunes. Es característicamente crónica y recurrente, y, a veces, indica el comienzo de las formas graves de esta micosis (Pappas, et al., 2009). Las levaduras del género Candida son microorganismos pertenecientes a la microbiota normal de individuos sanos, principalmente en la mucosa oral, el tracto gastrointestinal y el tracto genitourinario femenino (Shao, et al., 2007). Sin embargo, estos hongos son responsables de diferentes manifestaciones clínicas, especialmente en pacientes inmunocomprometidos, que van desde infecciones de la piel y mucosas a infecciones sistémicas (Sardi, et al., 2013). Su importancia viene de la alta frecuencia con que colonizan e infectan el huésped humano (De Bernardis, et al., 2004), siendo el cuarto patógeno más común asociado con los casos de infección nosocomial (Wisplinghoff, et al., 2004)...

Human Papillomavirus (HPV): Making the case for “Immunisation for All”

Human Papillomavirus (HPV) contributes to the most common sexually transmitted infections, with repeated and persistent infection with particular types causing disease in both men and women. Infection with low-risk HPV types can lead to genital warts and benign lesions of the oral cavity, while high-risk types can cause various HPV-related malignancies. The incidence of head and neck cancer has been rising in the past number of decades mostly due to oropharyngeal cancer linked to HPV infection. HPV vaccination has been shown to be effective for cervical and other anogenital HPV-related cancers, and there is significant potential for HPV vaccination to prevent oropharyngeal cancers, given that the HPV types implicated in this disease can be protected against by the HPV vaccine. Few countries have implemented a universal HPV vaccination programme for males and females, with many countries arguing that female only vaccination programmes protect males via herd immunity, and that men-who-have-sex-with-men will be protected via targeted vaccination programmes. We argue these may be limited in their effectiveness. We propose that the most effective, practical, ethical and potentially cost effective solution is universal HPV vaccination that might lead to control of HPV-related diseases in men and women alike.

Erosão dentária associada a distúrbios alimentares

Os distúrbios alimentares traduzem-se em comportamentos, quer pela falta, quer pelo excesso da ingestão de alimentos, sendo a anorexia nervosa e a bulimia as que mais frequentemente se verificam na população. O crescente aumento destes distúrbios, nomeadamente a obsessão pelos corpos magros, está relacionada com o impacto que os media têm na sociedade, uma vez que a magreza é vista como mecanismo de atracção sexual e de integração social. O diagnóstico das desordens alimentares não é fácil de ser efectuado, dado que os sinais são muitas vezes omitidos/ocultados pelos indivíduos. Os médicos dentistas podem ter um papel importante na sua detecção, dadas as manifestações precoces das alterações alimentares na cavidade oral. Há diversos sintomas comuns aos dois distúrbios alimentares como a erosão dentária, a hipersensibilidade dentinária, a hipertrofia das glândulas salivares e consequente hipossalivação, a cárie dentária, a doença periodontal, as mucosites, a candidíase oral e a queilite angular. Resultante do vómito induzido, há o aporte de ácido proveniente do conteúdo gástrico que induz alterações estruturais do esmalte e dentina, facilitando o processo de desgaste erosivo. Os fenómenos erosivos são uma das manifestações orais mais evidentes dos distúrbios alimentares. O conhecimento dos sinais, sintomas e da forma de evolução da erosão dentária, é imprescindível, e acaba por diferenciar a atuação profissional que possibilita um diagnóstico eficaz e o tratamento correto. A elaboração desta dissertação tem como objectivo reforçar a informação sobre estes fenómenos para que possam ser mais eficazmente prevenidos, diagnosticados e controlados/tratados. Para tal efectuou-se uma pesquisa na B-On, Medline/PubMed, sciELO, RCAAP e em livros, de informação válida sobre o tema. Interpôs-se limitação temporal e usaram-se as seguintes palavras-chave na seleção de artigos: “Dental erosion”, “Erosive wear”, “Anorexia”, “Bulimia”, “Eating disorders”.

Tumores do espaço parafaríngeo: experiência de 10 anos do IPO-LFG

Introdução: As neoplasias do espaço parafaríngeo são raras, representando apenas 0,5% dos tumores da cabeça e pescoço. A maioria são benignas, mas uma ampla variedade de patologias benignas e malignas podem ser encontradas neste espaço, o que cria desafios complexos de diagnóstico e tratamento. Objetivo: Descrever e analisar uma série de casos de neoplasias primárias do espaço parafaríngeo tratadas no Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG). Material e métodos: Estudo retrospetivo, com recolha e análise dos dados dos processos clínicos de tumores primários do espaço parafaríngeo, que foram diagnosticados ou referenciados ao IPOLFG entre 1 de Janeiro de 2003 e 31 de Dezembro de 2013. Resultados: Foram incluídos 38 doentes. A idade mediana foi de 52 anos (Âmbito Interquartil: 40-63 anos). Dez (26,3%) doentes eram assintomáticos. O sintoma mais comum à apresentação foi a sensação de corpo estranho orofaríngeo (23,7%) e o achado mais frequente foi um abaulamento orofaríngeo (78,4%). Todos os doentes fizeram exames de imagem pré-operatórios: 94,7% tomografia computorizada e 68,4% ressonância magnética. A citologia aspirativa foi realizada em 39,5%. 31 tumores eram benignos (81,6%), sendo os mais frequentes os adenomas pleomórficos (58,1%). 7 eram malignos (18,4%), com os carcinomas exadenomas pleomórficos (28,6%) e os linfomas (28,6%) sendo os mais comuns. 36 doentes (94,7%) foram submetidos a tratamento cirúrgico primário; os outros 2 doentes (5,3%) receberam tratamento não cirúrgico, com quimioterapia e quimioradioterapia, respectivamente. A abordagem cervical foi a mais utilizada (80%). A mandibulotomia foi necessária em apenas 5,7%. A complicação mais frequente foi a neuropatia de pares cranianos de novo, identificada em 22,2%. Destes, 75% foram sequela da resseção de tumores neurogénicos. Todas as neuropatias que resultaram da resseção de tumores não neurogénicos foram transitórias. O follow-up mediano foi de 6,5 anos. A taxa de recorrência foi de 13,5%. Conclusões: Os tumores do espaço parafaríngeo requerem um elevado índice de suspeição para serem diagnosticados num estadio precoce. A resseção cirúrgica completa é o principal tratamento. A abordagem cirúrgica deve ser selecionada caso a caso, mas a cervical fornece um excelente acesso à maioria dos tumores deste espaço

Frecuencia, manifestaciones clínicas y de laboratorio de infecciones respiratorias en pacientes con V.I.H./S.I.D.A. del Hospital Vicente Corral Moscoso de la ciudad de Cuenca período 2001-2007

Estudio descriptivo, se revisó y recolectó la información necesaria obtenidas de las historias clínicas de todos los pacientes diagnosticados de V.I.H.(+) en el departamento de clínica. Para el análisis estadístico: se utilizó los programas S.P.S.S. versión 10.0 para Windows y Microsoft Excel 2007, en la presentación de los datos se utilizó cuadros y gráficos en correspondencia con el tipo de variable y asociación que se deseó. Resultados: de un total de 7709 hospitalizados, 200 pacientes se encontraron infectados con V.I.H.(+), de los cuales 81 pacientes presentaron afección respiratoria, el 78de estos correspondió a los hombres, la mayoría se encontró entre los 26-35 años, la mortalidad se presentó en el 27,16; en los datos demográficos se encontró que los residentes de la sierra, católicos, solteros, un tipo de educación secundaria, la actividad manual sobresalían en los grupos de los pacientes; las principales patologías encontradas fueron Candidiasis Orofaríngea, Neumonía por P. Jirovecii, Tuberculosis Pulmonar y Neumonía Adquirida en la Comunidad. Se encontraron patrones en Radiología y Linfocitos CD4+ según principales patologías respiratorias. Conclusiones: el sexo masculino es el más frecuente dentro de los 81 pacientes V.I.H. que presentaron patologías respiratorias en el área de clínica, donde el sexo masculino fue el más frecuente, se presentó mortalidad alta en las primeras dos semanas de hospitalización, los datos demográficos son semejantes a nuestra región, la Candidiasis Oral y neumonía por P. Jirovecii fueron las principales afecciones en vías aéreas superior e inferior respectivamente, se encontró relación entre características radiológicas y linfocitos C.D.4+ asociadas con infecciones respiratorias en pacientes con VIH/SIDA

Aislamiento e identificación de Candida albicans de la cavidad oral, mediante el uso del Agar cromogénico en la población interna del Asilo San Antonio y Casa de la Misericordia de la ciudad de San Miguel

La investigación se realizó en la población con los adultos mayores del Asilo San Antonio y Casa de la Misericordia de la Ciudad de San Miguel, en el período de Junio a Julio de 2015. El Objetivo de la investigación fue aislar e identificar Candida albicans en muestras de la cavidad oral, mediante el uso del agar cromogénico de la población interna del Asilo San Antonio y Casa de la Misericordia de la Ciudad de San Miguel. El diseño metodológico es de tipo descriptivo, prospectivo, transversal, y de laboratorio, para el cual se tomaron 61 muestras de la cavidad oral de los internos en el Asilo San Antonio y Casa de la Misericordia de la Ciudad de San Miguel a través de un hisopado bucal con el que se realizó un examen directo al fresco con Solución Salina estéril al 0.85% en la búsqueda de levaduras; posteriormente se procedió a sembrar en el agar cromogénico Brilliance Candida, para observar el crecimiento de colonias verdes, las cuales indican la presencia de Candida albicans, como agente causal de Candidiasis oral. Resultados: De las 61 muestras procesadas a 26 se les aisló e identificó Candida albicas con un porcentaje de 42.7%. De los 22 adultos mayores que presentaban lesiones sugestivas a candidiasis oral a 20 se les aisló e identificó Candida albicans de los cuales 15 (88.2%) pertenecen al Asilo San Antonio y 5 (100%) pertenecen a la Casa de la Misericordia. De los 10 adultos mayores que reportaron que no practican el aseo bucal se les aisló e identificó Candida albicans a 3 (33.3%) de ellos que se encuentran en el Asilo San Antonio. A 5 internos que reportaron practicar algunas veces el aseo bucal también se les aisló e identificó Candida albicans, 4 (44.4%) pertenecen al Asilo San Antonio y 1 (50%) a la Casa de la Misericordia. De los 18 adultos mayores que usan prótesis dentales a 7 (53.8%) se les aisló e identificó Candida albicans en el Asilo San Antonio y en la Casa de la Misericordia a 5 (100%) internos se les aisló e identificó Candida albicans. De 35 adultos mayores que reportaron que usan antibiótico a 14 (46.7%) que se encuentran en el Asilo San Antonio se les aisló e identificó Candida albicans. Con respecto Casa de la Misericordia de los 5 (100%) que reportaron que usan antibiótico no se les aisló Candida albicans. En 16 (26.2%) muestras no se observó crecimiento de ninguna especie de Candida. Conclusiones: Se estudió a la población interna del Asilo San Antonio y Casa de La Misericordia debido a que los adultos mayores son vulnerables a las infecciones por hongos oportunistas ya que su sistema inmunológico se encuentra disminuido unido a una serie de factores predisponentes, como el uso de prótesis dentales, antibióticos, mal higiene bucal y diabetes. Mediante el uso del agar Brilliance Candida se logró diferenciar otras especies del género Candida como Candida tropicalis 26.2%, Candida krusei 3.3% y Candida glabrata 1.6%. De acuerdo a estos resultados y las conclusiones de la investigación se plantean algunas recomendaciones orientadas principalmente al personal de salud para brindar apoyo a este tipo de estudios.

Candidaemia or Candidasis: Controversy of Staphylococcus Sexually Transmitted Infection?

Herbal medications are becoming increasingly popular but a most-extraordinary claim by traditional/herbal medical practitioners relates to a Gram-positive bacterium, Staphylococcus , which has been depicted as a deadly sexually transmitted disease that manifest in the form of worms and other symptoms; with contributory roles including infertility, sexual dysfunction and impotency. They further boasted that they are the only ones that possessed the remedy (herbal) for the Staphylococcus sexually transmitted scourge. In the absence of distinguishing phenotypic taxonomic tools, Staphylococcus and Candida spp. may be confused for each other. However, Staphylococcus is a bacterium and not an infection; therefore, there must be more to the traditional medical practitioners’ boasts in ability to cure an infection that was not an infection in the first place. In conclusion, the common sense is that candiaemia or candidiasis is most likely the misdiagnosed sexually transmitted Staphylococcus disease, which is of significant human clinical health issue.

Estudio de la patogenicidad de Candida y de la actividad de los fármacos antifúngicos en un modelo animal en Caenorhabditis elegans

171 p.

Review: Head and neck squamous cell carcinoma in sub-Saharan Africa

Aim Review the literature from 1990 to 2013 to determine known anatomic sites, risk factors, treatments, and outcomes of head and neck squamous cell carcinoma (HNSCC) in sub-Saharan Africa. Methods Using a systematic search strategy, literature pertaining to HNSCC in sub-Saharan Africa was reviewed and patient demographics, anatomic sites, histology, stage, treatment, and outcomes were abstracted. The contributions of human immunodeficiency virus (HIV), human papillomavirus (HPV) and behavioural risk factors to HNSCC in the region were assessed. Results Of the 342 papers identified, 46 were utilized for review, including 8611 patients. In sub-Saharan Africa, the oropharyngeal/oral cavity was found to be the most common site, with 7750 cases (90% of all cases). Few papers distinguished oropharyngeal from oral cavity, making identification of possible HPV-associated oropharyngeal squamous cell carcinoma (SCC) difficult. SCC of the nasopharynx, nasal cavity, or paranasal sinuses was identified in 410 patients (4.8% of all cases). Laryngeal SCC was found in 385 patients (4.5% of all cases), and only 66 patients (0.8% of all cases) with hypopharyngeal SCC were identified. In 862 patients with data available, 43% used tobacco and 42% used alcohol, and reported use varied widely and was more common in laryngeal SCC than that of the oropharyngeal/oral cavity. Toombak and kola nut use was reported to be higher in patients with HNSCC. Several papers reported HIV-positive patients with HNSCC, but it was not possible to determine HNSCC prevalence in HIV-positive compared to negative patients. Reports of treatment and outcomes were rare. Conclusions The oropharyngeal/oral cavity was by far the most commonly reported site of HNSCC reported in sub-Saharan Africa. The roles of risk factors in HNSCC incidence in sub-Saharan Africa were difficult to delineate from the available studies, but a majority of patients did not use tobacco and alcohol.

Polyphasic characterisation of Burkholderia cepacia complex species isolated from children with cystic fibrosis

Cystic fibrosis (CF) patients with Burkholderia cepacia complex (Bcc) pulmonary infections have high morbidity and mortality. The aim of this study was to compare different methods for identification of Bcc species isolated from paediatric CF patients. Oropharyngeal swabs from children with CF were used to obtain isolates of Bcc samples to evaluate six different tests for strain identification. Conventional (CPT) and automatised (APT) phenotypic tests, polymerase chain reaction (PCR)-recA, restriction fragment length polymorphism-recA, recA sequencing, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) were applied. Bacterial isolates were also tested for antimicrobial susceptibility. PCR-recA analysis showed that 36 out of the 54 isolates were Bcc. Kappa index data indicated almost perfect agreement between CPT and APT, CPT and PCR-recA, and APT and PCR-recA to identify Bcc, and MALDI-TOF and recA sequencing to identify Bcc species. The recA sequencing data and the MALDI-TOF data agreed in 97.2% of the isolates. Based on recA sequencing, the most common species identified were Burkholderia cenocepacia IIIA (33.4%), Burkholderia vietnamiensis (30.6%), B. cenocepacia IIIB (27.8%), Burkholderia multivorans (5.5%), and B. cepacia (2.7%). MALDI-TOF proved to be a useful tool for identification of Bcc species obtained from CF patients, although it was not able to identify B. cenocepacia subtypes.

First description of Candida nivariensis in Brazil: antifungal susceptibility profile and potential virulence attributes

This study evaluated the antifungal susceptibility profile and the production of potential virulence attributes in a clinical strain of Candida nivariensis for the first time in Brazil, as identified by sequencing the internal transcribed spacer (ITS)1-5.8S-ITS2 region and D1/D2 domains of the 28S of the rDNA. For comparative purposes, tests were also performed with reference strains. All strains presented low planktonic minimal inhibitory concentrations (PMICs) to amphotericin B (AMB), caspofungin (CAS), and voriconazole. However, our strain showed elevated planktonic MICs to posaconazole (POS) and itraconazole, in addition to fluconazole resistance. Adherence to inert surfaces was conducted onto glass and polystyrene. The biofilm formation and antifungal susceptibility on biofilmgrowing cells were evaluated by crystal violet staining and a XTT reduction assay. All fungal strains were able to bind both tested surfaces and form biofilm, with a binding preference to polystyrene (p < 0.001). AMB promoted significant reductions (≈50%) in biofilm production by our C. nivariensis strain using both methodologies. This reduction was also observed for CAS and POS, but only in the XTT assay. All strains were excellent protease producers and moderate phytase producers, but lipases were not detected. This study reinforces the pathogenic potential of C. nivariensis and its possible resistance profile to the azolic drugs generally used for candidiasis management.

Candida albicans Modifies the Protein Composition and Size Distribution of THP1 macrophages-derived Extracellular Vesicles.

The effectiveness of macrophages in the response to systemic candidiasis is crucial to an effective clearance of the pathogen. The secretion of proteins, mRNAs, non-coding RNAs and lipids through extracellular vesicles (EVs) is one of the mechanisms of communication between immune cells. EVs change their cargo to mediate different responses, and may play a role in the response against infections. Thus, we have undertaken the first quantitative proteomic analysis on the protein composition of THP1 macrophages-derived EVs during the interaction with Candida albicans. This study revealed changes in EVs sizes and in protein composition, and allowed the identification and quantification of 717 proteins. Of them, 133 proteins changed their abundance due to the interaction. The differentially abundant proteins were involved in functions relating to immune response, signaling, or cytoskeletal reorganization. THP1-derived EVs, both from control and from Candida-infected macrophages, had similar effector functions on other THP1-differenciated macrophages, activating ERK and p38 kinases, and increasing both the secretion of proinflammatory cytokines and the candidacidal activity; while in THP1 non-differenciated monocytes, only EVs from infected macrophages increased significantly the TNF-α secretion. Our findings provide new information on the role of macrophage-derived EVs in response to C. albicans infection and in macrophages communication.

Estudio proteómico de las vesículas extracelulares y del secretoma de "Candida albicans" y análisis funcional de proteínas de superficie celular

Candida albicans es un importante patógeno oportunista en humanos, que puede causar distintos tipos de infecciones, desde micosis superficiales hasta sistémicas. La candidiasis invasiva es una enfermedad que puede causar mortalidad en pacientes inmunocomprometidos. Para causar daño en el hospedador, C. albicans cuenta con una serie de factores de virulencia. Entre ellos destaca la capacidad de cambiar su forma de crecimiento de levadura a hifa. La superficie celular es la estructura más externa de la célula y el punto de contacto entre el hongo y el hospedador. Las proteínas de superficie tienen un papel importante en la integridad estructural de la célula y en la adherencia e invasión de células del hospedador. Una de las proteínas localizadas en la superficie celular es Ecm33, una proteína de pared celular con anclaje glicosilfosfatidilinositol (GPI). La deleción de esta proteína afecta a la morfología tanto de levaduras como de hifas, dando como resultado células con la pared celular alterada y virulencia reducida tanto en condiciones in vitro como in vivo. El secretoma o las proteínas secretadas por C. albicans son también relevantes en la interacción patógeno-hospedador. C. albicans secreta muchas proteínas importantes relacionadas con diferentes procesos, entre los que se incluyen la formación de biofilms, la adquisición de nutrientes y el mantenimiento de la integridad de la pared celular. Muchas de estas proteínas secretadas, como las pertenecientes a las familias de aspartil proteasas (Sap) y la familia de fosfolipasas B (Plb), también han sido detectadas en la pared celular, ya que deben pasar a través de ella en su tránsito hacia el medio extracelular. Estas proteínas tienen un péptido señal en el extremo N-terminal que es el responsable de dirigirlas a la ruta clásica de secreción. Sin embargo, cerca de un tercio de las proteínas identificadas en el medio extracelular de C. albicans no poseen dicho péptido señal en su secuencia...

Interrogation of human monoclonal antibodies induced by 4C-MenB to identify protective antigens contained in the OMV component

Neisseria meningitidis is a gram negative human obligated pathogen, mostly found as a commensal in the oropharyngeal mucosa of healthy individuals. It can invade this epithelium determining rare but devastating and fast progressing outcomes, such as meningococcal meningitidis and septicemia, leading to death (about 135000 per year worldwide). Conjugated vaccines for serogroups A, C, W135, X and Y were developed, while for N. meningitidis serogroup B (MenB) the vaccines were based on Outern Membrane Vesicles (OMV). One of them is the 4C-MenB (Bexsero). The antigens included in this vaccine’s formulation are, in addition to the OMV from New Zeland epidemic strain 98/254, three recombinant proteins: NadA, NHBA and fHbp. While the role of these recombinant components was deeply characterized, the vesicular contribution in 4C-MenB elicited protection is mediated mainly by porin A and other unidentified antigens. To unravel the relative contribution of these different antigens in eliciting protective antibody responses, we isolated human monoclonal antibodies (mAbs) from single-cell sorted plasmablasts of 3 adult vaccinees peripheral blood. mAbs have been screened for binding to 4C-MenB components by Luminex bead-based assay. OMV-specific mAbs were purified and tested for functionality by serum bactericidal assay (SBA) on 18 different MenB strains and characterized in a protein microarray containing a panel of prioritized meningococcal proteins. The bactericidal mAbs identified to recognize the outer membrane proteins PorA and PorB, stating the importance of PorB in cross-strain protection. In addition, RmpM, BamE, Hyp1065 and ComL were found as immunogenic components of the 4C-MenB vaccine.

Longitudinal Micro CT-driven biomarkers as a tool to evaluate lung fibrosis progression and antifibrotic efficacy in a refined Bleomycin mouse model

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive disease with no curative pharmacological treatment. Animal models play an essential role in revealing molecular mechanisms involved in the pathogenesis of the disease. Bleomycin (BLM)-induced lung fibrosis is the most widely used and characterized model for anti-fibrotic drugs screening. However, several issues have been reported, such as the identification of an optimal BLM dose and administration scheme as well as gender-specificity. Moreover, the balance between disease resolution, an appropriate time window for therapeutic intervention and animal welfare remains critical aspects yet to be fully elucidated. In this thesis, Micro CT imaging has been used as a tool to identify the ideal BLM dose regimen to induce sustained lung fibrosis in mice as well as to assess the anti-fibrotic effect of Nintedanib (NINT) treatment upon this BLM administration regimen. In order to select the optimal BLM dose scheme, C57bl/6 male mice were treated with BLM via oropharyngeal aspiration (OA), following either double or triple BLM administration. The triple BLM administration resulted in the most promising scheme, able to balance disease resolution, appropriate time-window for therapeutic intervention and animal welfare. The fibrosis progression was longitudinally assessed by micro-CT every 7 days for 5 weeks after BLM administration and 5 animals were sacrificed at each timepoint for the BALF and histological evaluation. The antifibrotic effect of NINT was assessed following different treatment regimens in this model. Herein, we have developed an optimized mouse model of pulmonary fibrosis, enabling three weeks of the therapeutic window to screen putative anti-fibrotic drugs. micro-CT scanning, allowed us to monitor the progression of lung fibrosis and the therapeutical response longitudinally in the same subject, drastically reducing the number of animals involved in the experiment.