Desenvolvimento e validação de método bioanalítico por difusão em ágar para determinação da potência do fenticonazol creme vaginal

O nitrato de fenticonazol é um derivado imidazólico de amplo espectro de ação contra patógenos dermatófitos, como Malassezia furfur e leveduras, a principal delas Candida albicans. É necessário que se possa garantir a qualidade dos medicamentos produzidos. O controle de qualidade é uma ferramenta que assegura a qualidade, eficácia e segurança desses produtos até o momento de sua utilização e faz-se necessário que métodos analíticos sejam desenvolvidos e validados para o doseamento desses fármacos. A grande importância dos métodos microbiológicos utilizados no controle de qualidade de fármacos com atividade antimicrobiana é demonstrar sua atividade biológica por meio de seu efeito inibitório no crescimento de micro-organismo. A diminuição da atividade antimicrobiana ou potência pode indicar alguma alteração sutil na molécula, que é pouco detectável em um método analítico físico-químico. O objetivo deste trabalho foi desenvolver e validar o método microbiológico difusão em ágar para o produto nitrato de fenticonazol creme vaginal. O método de difusão em ágar foi desenvolvido utilizando meio de cultura ágar Sabouraud e cepa de Candida albicans ATCC 64548 como micro-organismo teste. Para o ensaio foi utilizado o delineamento 3 x 3, conforme preconiza a Farmacopeia Brasileira (2010). A validação do método proposto demonstrou linearidade entre os diâmetros dos halos de inibição e o logaritmo da concentração na faixa de 50,0 a 200,0 μg/mL. O método obteve resultados precisos e lineares com r2=1. A exatidão foi comprovada pelo intervalo de tolerância de 98,83%, dentro dos limites permitidos. O método foi validado de maneira satisfatória, podendo ser utilizado em análises do Controle de Qualidade deste fármaco.

Perfil de fatores de virulência das Escherichia coli isoladas do útero de vacas holandesas após o parto e sua relação com a microbiota uterina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Performance da contração muscular do assoalho pélvico de primíparas que realizaram parto vaginal ou parto cesárea

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Effect of short training on vaginal fluid microscopy (wet mount) learning

Objective: Is it feasible to learn the basics of wet mount microscopy of vaginal fluid in 10 hours?Materials and Methods: This is a pilot project wherein 6 students with different grades of education were invited for being tested on their ability to read wet mount microscopic slides before and after 10 hours of hands-on training. Microscopy was performed according to a standard protocol (Femicare, Tienen, Belgium). Before and after training, all students had to evaluate a different set of 50 digital slides. Different diagnoses and microscopic patterns had to be scored. kappa indices were calculated compared with the expert reading. Results: All readers improved their mean scores significantly, especially for the most important types of altered flora (p < .0001). The mean increase in reading concordance (kappa from 0.64 to 0.75) of 1 student with a solid previous experience with microscopy did not reach statistical significance, but the remaining 5 students all improved their scores from poor performance (all kappa < 0.20) to moderate (kappa = 0.53, n = 1) to good (kappa > 0.61, n = 4) concordance. Reading quality improved and reached fair to good concordance on all microscopic items studied, except for the detection of parabasal cells and cytolytic flora. Conclusions: Although further improvement is still possible, a short training course of 10 hours enables vast improvement on wet mount microscopy accuracy and results in fair to good concordance of the most important variables of the vaginal flora compared to a reference reader.

Immunocytochemistry associated with oral exfoliative cytology methodological analysis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evaluation of dentists' knowledge of the use of oral exfoliative cytology in clinical practice

Oral exfoliative cytology (EC) is a rapid and practical complementary test that can be used in the diagnosis of various diseases. The objectives of this study were to assess dentists'knowledge of EC and to disseminate information on the technique. The study included 240 dentists from the city of Anapolis (Goias, Brazil) who answered a questionnaire regarding demographic data and data relating to EC and who then received information on the technique. The data revealed that 26.92% of the dentists did not know about EC. Among those who reported knowledge of EC, 33.91% did not know the procedures for performing the technique, and 85.38% had no experience with EC. In addition, 54.09% of the answers regarding the instrument used to collect the material and 56.82% of the answers concerning suitable fixatives were inappropriate. Although 73.02% of the dentists adequately identified the pathologies for which EC is recommended, it was concluded that the respondents had poor knowledge of EC. There is a need to increase dentists'awareness of this subject and of EC's use in diagnostic practices.

Primary squamous cell carcinoma of the thyroid diagnosed as anaplastic carcinoma: failure in fine-needle aspiration cytology?

A case of primary squamous-cell carcinoma (SCC) of the thyroid which had been initially diagnosed as an anaplastic carcinoma (ATC) is described: female, 73 years old, with a fast-growing cervical nodule on the left side and hoarseness for 3 months. Ultrasonography showed a 4.5 cm solid nodule. FNA was compatible with poorly differentiated carcinoma with immunoreactivity for AE1/AE3, EMA. Thyroidectomy was performed. Histopathological examination showed a nonencapsulated tumor. Immunohistochemistry disclosed positivity for AE1/AE3, p53,p63, and Ki67. The diagnosis was ATC. A second opinion reported tumor consisting of squamous cells, with intense inflammatory infiltrate both in tumor and in the adjacent thyroid, with final diagnosis of SCC, associated with Hashimoto thyroiditis. No other primary focus of SCC was found. Patient has shown a 48-month survival period. Clinically, primary SCCs of the thyroid and ATCs are similar. The distinction is often difficult particularly when based on the cytological analysis of FNA material.

A curcumin-loaded liquid crystal precursor mucoadhesive system for the treatment of vaginal candidiasis

Women often develop vaginal infections that are caused primarily by organisms of the genus Candida. The current treatments of vaginal candidiasis usually involve azole-based antifungals, though fungal resistance to these compounds has become prevalent. Therefore, much attention has been given to molecules with antifungal properties from natural sources, such as curcumin (CUR). However, CUR has poor solubility in aqueous solvents and poor oral bioavailability. This study attempted to overcome this problem by developing, characterizing, and evaluating the in vitro antifungal action of a CUR-loaded liquid crystal precursor mucoadhesive system (LCPM) for vaginal administration. A low-viscosity LCPM (F) consisting of 40% wt/wt polyoxpropylene-(5)-polyoxyethylene-(20)-cetyl alcohol, 50% wt/wt oleic acid, and 10% wt/wt chitosan dispersion at 0.5% with the addition of 16% poloxamer 407 was developed to take advantage of the lyotropic phase behavior of this formulation. Notably, F could transform into liquid crystal systems when diluted with artificial vaginal mucus at ratios of 1:3 and 1:1 (wt/wt), resulting in the formation of F30 and F100, respectively. Polarized light microscopy and rheological studies revealed that F behaved like an isotropic formulation, whereas F30 and F100 behaved like an anisotropic liquid crystalline system (LCS). Moreover, F30 and F100 presented higher mucoadhesion to porcine vaginal mucosa than F. The analysis of the in vitro activity against Candida albicans revealed that CUR-loaded F was more potent against standard and clinical strains compared with a CUR solution. Therefore, the vaginal administration of CUR-loaded LCPMs represents a promising platform for the treatment of vaginal candidiasis.

Avaliação proteômica do conteúdo vaginal em resposta ao tratamento da vaginose bacteriana

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Maturidade pulmonar e fetal de bezerros da raça holandesa preta e branca tecém-nascidos, a termo e hígidos: análise do líquido amniótico

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Carcinoma de células escamosas em assoalho vaginal de um bovino nelore associado a metástase em pulmão: relato de caso

The squamous cell carcinoma (SCC) is a neoplasm that affect pets and production animals and it’s very common in tropical countries like Brazil; develops in sparsely pigmented, stratified squamous epithelium and in mucosal surfaces exposed Ultraviolet action. The SCC is quite infiltrative but rarely causes metastases. Its occurrence in the female reproductive tract is recognized in the literature on cattle breeds from Europe. This case was a female bovine, Nelore, adult, who was referred to the "Hospital Veterinário Luiz Quintiliano de Oliveira" with dark brown fluid leakage and putrid odor, associated with ulcerative growth of the vagina. Because the extent of injury, the animal was euthanized and taken to the necropsy, which was observed on board structure, ulcerated and pus in the vaginal floor, infiltrated into the pelvic cavity to the serosa of the uterine body. The microscopic findings were detected neoplastic proliferation of epithelial cells in the floor and vaginal metastases in the lung parenchyma, and classified the primary tumor and metastasis as squamous cell carcinoma moderately differentiated.

Fine needle aspiration cytology of lobular breast carcinoma and its variants

Objective: To identify associations between cytological criteria in fine needle aspiration (FNA) specimens and histological subtypes of lobular breast carcinoma (classical and other types). Study Design: FNA cytology and mastectomy specimens from 72 cases of invasive lobular breast carcinoma were consecutively retrieved from the files of the Amaral de Carvalho Hospital, Jaú-São Paulo, Brazil. All cases were reviewed regarding five cytological criteria: cellularity, cellular cohesion, presence of inflammation, nucleoli and nuclear atypia. The χ2 test or Fisher's exact tests with 95% confidence intervals (CI) were used. Results: The classical type showed lower initial cytological diagnosis of malignancy compared to the other variants (p = 0.017; odds ratio (OR) 0.26, 95% CI 0.89-0.80). Moderate/intense cellular cohesion (p = 0.011; OR 0.18, 95% CI 0.04-0.73) and mild atypia (p = 0.000; OR 16.15, 95% CI 3.20-81.48) were significantly associated with the classical type of lobular breast carcinoma, while the absence of inflammation (p = 0.082; OR 0.36, 95% CI 0.12-1.15) was marginally associated with the classical type. Conclusions: In cytology, the characterization of lobular carcinoma as malignant is difficult, especially the classical type. The association between cell cohesion and the classical type of lobular breast carcinoma may be one of the factors that complicate this diagnosis.

Correlação entre a força muscular do assoalho pélvico e status hormonal da vagina em mulheres continentes

Pós-graduação em Bases Gerais da Cirurgia - FMB

Cytology of tracheobronchial and bronchoalveolar lavage in healthy Holteins calves during the first month of life

Benesi F.J., Wachholz L., Bertagnon H.G., Leal M.L.R., Mori E. & Fernandes W.R. 2012. [Cytology of tracheobronchial and bronchoalveolar lavage in healthy Holteins calves during the first month of life.] Citologia dos lavados traqueobronquico (LTB) e broncoalveolar (LBA) de bezerros holandeses sadios durante o primeiro mes de vida. Pesquisa Veterinaria Brasileira 32(3):267-270. Departamento de Clinica Medica, Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Sao Paulo, SP 05508-270, Brazil. E-mail: febencli@usp.br The neonatal calf is a critical moment for adaptation of the newborn to extra uterine life. The respiratory tract is functionally very demanded and often affected by disease, resulting in direct loss of their function and causing serious economic losses in livestock. The basic point to reduce these losses is appropriate clinical evaluation of neonates; but the diagnosis based solely in physical examination is very difficult to establish. The use of complementary analysis such cytology of the respiratory tract becomes an important diagnostic tool; however their findings must be standardized in the face of different techniques employed. This research studied the dynamics of the cellularity of the bronchoalveolar and tracheobronchial region obtained through lung lavage harvested by nasotracheal catheterization technique and tracheocenthesis respectively, during the first month of life of healthy calves. The tracheobronchial cytology was influenced by the time, showing decreased number of alveolar macrophages and greater number of neutrophils, possibly increased by local irritation caused by the technique, which was repeated sequentially, and/or through greater stimulation of inhaled microorganisms deposited in this region. In the bronchoalveolar region no variation in the cellular constituents in function of time was found. The results allowed the conclusion the cell population of the tracheobronchial region has changed over the week-old calves, possibly due to the technique used and/or to the normal region physiology, represented by higher magnitudes of neutrophils. Otherwise, the cells of the broncholaveolar region showed a stable behavior during the first month of life of newborn calves, presenting numerical predominance of alveolar macrophages. O período neonatal dos bezerros é um momento crítico para adaptação do recém-nascido à vida extra uterina e o sistema respiratório, um dos mais exigidos funcionalmente, é frequentemente afetado por enfermidades, redundando no prejuízo direto da sua função e acarretando perdas econômicas importantes na pecuária. O ponto básico para reduzir estas perdas, é representado pela adequada avaliação clínica dos neonatos, todavia o diagnóstico baseado exclusivamente no exame ísico é muito di ícil de ser estabelecido. O uso de exames complementares como a citologia do trato respiratório torna-se uma ferramenta diagnóstica importante nestes casos, porém faz-se necessário, padronizar seus achados frente às diferentes técnicas empregadas para a sua obtenção. Assim, o presente estudo propôs-se acompanhar as variações dos constituintes celulares da região traqueobrônquica e broncoalveolar obtidos por lavados respiratórios pelos métodos de traqueocentese e por colheita nasotraqueal respectivamente, durante o primeiro mês de vida de bezerros sadios. Observou-se alteração no quadro citológico ao longo do tempo, quando a região traqueobrônquica foi lavada, expresso por diminuição da porcentagem de macrófagos alveolares, com aumento de neutró ilos, possivelmente, por maior irritação local provocada pela técnica, que se repetiu sequencialmente e/ou por maior estimulo de microorganismos inalados depositados nesta região. Na região broncoalveolar, não encontraram- -se variações nos constituintes celulares em função do tempo. Os resultados permitiram a conclusão que a população celular da região traqueobrônquica modi icou-se ao longo das semanas de vida dos bezerros, possivelmente pela técnica empregada e/ou isiologia normal da região, sendo representadas por maiores magnitudes de neutró ilos. De modo diverso, na região broncolaveolar, as células evidenciaram um comportamento estável durante o primeiro mês de vida dos bezerros neonatos, apresentando predomínio numérico dos macrófagos alveolares.