Stability Testing of Spray- and Spouted Bed-Dried Extracts of Passiflora alata

The aim of this research was to perform a stability testing of spray- and spouted bed-dried extracts of Passiflora alata Dryander (Passion flower) under stress storage conditions. Spouted bed- and spray-dried extracts were characterized by determination of the average particle diameter (dP), apparent moisture content (XP), total flavonoid content (TF), and vitexin content. Smaller and more irregular particles were generated by the spouted bed system due to a higher attrition rate (surface erosion) inside the dryer. The SB dryer resulted in an end product with higher concentration of flavonoids (approximate to 10%) and lower moisture content (1.6%, dry basis) than the spray dryer, even with both dryers working at similar inlet drying air temperature and ratio between the extract feed flow rate to drying air flow rate (Ws/Wg). Samples of the spouted bed- and spray-dried extracts were stored at two different temperatures (34 and 45 degrees C) and two different relative humidities (52 and 63% RH for 34 degrees C; 52 and 60% RH for 45 degrees C) in order to perform the stability testing. The dried extracts were stored for 28 days and were analyzed every 4 days. The flavonoid vitexin served as the marker compound, which was assayed during the storage period. Results revealed shelf lives ranging from 9 to 184 days, depending on the drying process and storage conditions.

The assessment of oral cytology utilizing ploidy analysis and virtual microscopy for the early detection of epithelial dysplasia and neoplasia in oral mucosal lesions

Oral squamous cell carcinoma (OSCC) is associated with high morbidity and mortality which is due, at least in part, to late detection. Precancerous and cancerous oral lesions may mimic any number of benign oral lesions, and as such may be left without investigation and treatment until they are well advanced. Over the past several years there has been renewed interest in oral cytology as an adjuvant clinical tool in the investigation of oral mucosal lesions. The purpose of the present study was to compare the usefulness of ploidy analysis after Feulgen stained cytological thin-prep specimens with traditional incisional biopsy and routine histopathological examination for the assessment of the pre-malignant potential of oral mucosal lesions. An analysis of the cytological specimens was undertaken with virtual microscopy which allowed for rapid and thorough analysis of the complete cytological specimen. 100 healthy individuals between 30 and 70 years of age, who were non-smokers, non-drinkers and not taking any medication, had cytological specimens collected from both the buccal mucosa and lateral margin of tongue to establish normal cytology parameters within a control population. Patients with a presumptive clinical diagnosis of lichen planus, leukoplakia or OSCC had lesional cytological samples taken prior to their diagnostic biopsy. Standardised thin preparations were prepared and each specimen stained by both Feuglen and Papanicolau methods. High speed scanning of the complete slide at 40X magnification was undertaken using the Aperio Scanscope TM and the green channel of the resultant image was analysed after threshold segmentation to isolate only nuclei and the integrated optical density of each nucleus taken as a gross measure of the DNA content (ploidy). Preliminary results reveal that ploidy assessment of oral cytology holds great promise as an adjunctive prognostic factor in the analysis of the malignant potential of oral mucosal lesions.

Relationship between induced sputum cytology and inflammatory status with lung structural and functional abnormalities in asbestosis

Background Asbestosis is associated with lung cellular and immunological abnormalities. Induced sputum cytology and local and systemic markers of inflammation may be helpful to characterize disease status and progression in these patients. Methods Thirty-nine ex-workers with asbestosis on high-resolution CT (HRCT) and 21 non-exposed controls were evaluated. Sputum cytology and IL-8 in serum and sputum were related to lung function impairment. Results Subjects with asbestosis had reduced sputum cellularity but higher macrophagel neutrophil ratio and % macrophage as compared with controls. Sputum and serum IL-8 were also higher in patients with asbestosis (P < 0.05). In addition, evidence of lung architectural distorption on HRCT was associated with increased levels of serum IL-8. Interestingly, absolute macrophage number was negatively correlated with total lung capacity (r = -0.40; P = 0.04) and serum IL-8 to lung diffiusing capacity (r = -0.45; P = 0.01). Conclusions Occupationally exposed subjects with asbestosis on HRCT have cytologic abnormalities in induced sputum and increased local and systemic pro-inflammatory status which are correlated to functional impairment.

Anaplastic Thyroid Carcinoma Morphologic Findings and PAX-8 Expression in Cytology Specimens

Objective To evaluate the morphologic findings most encountered in anaplastic thyroid carcinomas (ATCs) and evaluate for the expression of PAX-8. Study Design The cytology specimens from 21 cases of ATC were evaluated for the following several cytologic criteria, cell morphology, pleomorphism, presence or absence of multinucleated cells, neutrophilic infiltrate and well-differentiated component. Immunohistochemical studies for PAX-8 were performed on cell blocks in selected cases. Results The most common morphology present was epithelioid, followed by spindle, multinucleated giant cell and rhabdoid, with 75% demonstrating more than 1 cell morphology. Marked pleomorphism (81%), neutrophilic infiltrate (90%) and necrosis (63%) were frequent. No nuclear grooves, colloid or well-differentiated component was idenrifled in any case. Immunocytochemical stains for PAX-8 were negative in 5 cases in which a cell block was available. Conclusion ATC is a tumor with diverse morphologic characteristics, and more than 1 cell morphology is usually present A neutrophilic infiltrate is a common finding and represents a clue to the correct diagnosis. (Acta Cytol 2010;54:668-672)

Peritoneal cytology: impact on disease-free survival in clinical stage I endometrioid adenocarcinoma of the uterus

The prognostic significance of positive peritoneal cytology in endometrial carcinoma has led to the incorporation of peritoneal cytology into the current FIGO staging system, While cytology was shown to be prognostically relevant in patients with stage II and III disease, conflicting data exists about its significance in patients who would have been stage I but were classified as stage III solely and exclusively on the basis of positive peritoneal cytology (clinical stage I). Analysis was based on the data of 369 consecutive patients with clinical stage I endometrioid adenocarcinoma of the endometrium. Standard treatment consisted of an abdominal total hysterectomy, bilateral salpingo-oophorectomy with or without pelvic lymph node dissection. Peritoneal cytology was obtained at laparotomy by peritoneal washing of the pouch of Douglas and was considered positive if malignant cells could be detected regardless of the number of malignant cells present. Disease-free survival (DFS) was considered the primary statistical endpoint. In 13/369 (3.5%) patients, positive peritoneal cytology was found. The median follow-up was 29 months and 15 recurrences occurred. Peritoneal cytology was independent of the depth of myometrial invasion and the grade of tumour differentiation, Patients with negative washings had a DFS of 96'7e at 36 months compared with 67% for patients with positive washings (log-rank P < 0.001). The presence of positive peritoneal cytology in patients with clinically stage I endometrioid adenocarcinoma of the endometrium is considered an adverse prognostic factor. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

Estimação da área foliar por método não destrutivo, utilizando medidas lineares das folhas de espécies de Passiflora

Objetivou-se, com este trabalho, desenvolver uma equação confiável, que possibilite calcular a área foliar de forma rápida e não destrutiva, utilizando medidas lineares da folha, para as diferentes espécies de Passiflora (P. alata, P. coccinea, P. gibertii, P. ligularis, P. misera, P. mucronata, P. nitida, P. setacea). Foram coletadas trezentas folhas, de diferentes tamanhos, de cada espécie. Realizaram-se análises de regressão da área foliar versus comprimento da nervura principal, maior largura da folha e o produto destas. A área foliar foi medida com um dispositivo de medição automática e as medidas lineares foram determinadas utilizando-se régua. O modelo linear escolhido, para todas as espécies, foi o que utiliza o produto das variáveis independentes, por apresentar maior coeficiente de determinação (R²) e maior significância do coeficiente de regressão. Os modelos apresentaram coeficiente de determinação ajustado superior a 0,927 e índice de desempenho ótimo, segundo a classificação de Camargo e Sentelhas. No processo de validação do modelo, mostrou-se que a correlação da área foliar medida com a área foliar observada foi muito alta. Os resultados obtidos neste estudo demonstram que a área foliar das espécies de Passifloras podem ser preditas, usando-se as determinações do comprimento e da largura das folhas.

Enraizamento adventício de estacas do maracujazeiro silvestre Passiflora mucronata Lam.: forma de veiculação e concentrações do ácido indol-3-butírico

Passiflora mucronata Lam. apresenta como características ser ornamental, medicinal e resistente à bacteriose nas folhas, além de ser altamente resistente à antracnose nos frutos e ramos. Objetivou-se, com este trabalho, avaliar a forma de veiculação, líquida ou sólida, e as concentrações do ácido indol-3-butírico (AIB) no enraizamento adventício de estacas de P. mucronata. O experimento foi conduzido no delineamento de blocos ao acaso, em esquema fatorial 2x5 (formas de veiculação do AIB: líquida e sólida x concentrações de AIB: 0; 250; 500; 750 e 1000 mg L-1, se líquido, ou mg Kg-1, se sólido), com quatro repetições, de 16 estacas cada. As características avaliadas foram enraizamento (%), número de raiz, comprimento da maior raiz (cm), volume de raiz (cm³) e massa de matéria seca de raiz (g). Verificou-se que o enraizamento de estacas de P. mucronata Lam. independe da forma como é veiculado o AIB, se sólida ou líquida, porém a qualidade do sistema radicular, avaliada por meio do número, volume e massa de matéria seca de raízes, foi superior à obtida ao se empregar o AIB sólido. O comportamento do enraizamento, do número e do comprimento de raízes foi linear crescente, até a máxima concentração de AIB (1000 mg L-1 ou mg Kg-1).

Optimization of immunocytochemistry in cytology: comparison of two protocols for fixation and preservation on cytospin and smear preparations

Objective: A new protocol for fixation and slide preservation was evaluated in order to improve the quality of immunocytochemical reactions on cytology slides. Methods: The quality of immunoreactions was evaluated retrospectively on 186 cytology slides (130 direct smears, 56 cytospins) prepared from different cytology samples. Ninety-three of the slides were air dried, stored at -20 °C and fixed in acetone for 10 minutes (Protocol 1), whereas the other 93 were immediately fixed in methanol at -20 °C for at least 30 minutes, subsequently protected with polyethylene glycol (PEG) and stored at room temperature (Protocol 2). Immunocytochemical staining, with eight primary antibodies, was performed on a Ventana BenchMark Ultra instrument using an UltraView Universal DAB Detection Kit. The following parameters were evaluated for each immunoreaction: morphology preservation, intensity of specific staining, background and counterstain. The slides were blinded and independently scored by four observers with marks from 0 to 20. Results: The quality of immunoreactions was better on methanol-fixed slides protected with PEG than on air-dried slides stored in the freezer: X¯ = 14.44 ± 3.58 versus X¯ = 11.02 ± 3.86, respectively (P < 0.001). Conclusion: Immediate fixation of cytology slides in cold methanol with subsequent application of PEG is an easy and straightforward procedure that improves the quality of immunocytochemical reactions and allows the storage of the slides at room temperature.

A morphological protocol and guide-list on uterine cervix cytology associated to Papillomavirus infection

The present study was designed to further assess the validity of the cytological description of morphological lesions said to be related to Papillomavirus (HPV) infections in senior women. The casuistic comprised 196 cervical smears from a group of women with no clinical or morphological evidence of neoplasia, collected simultaneously with samples submitted to detection of HPV DNA by PCR in a previous study. Three experienced cytologists studied each slide in two different conditions, with an interval of 20 months between them. The first approach was performed under routine laboratory standards, whereas the second was guided by a list of 16 well-defined parameters indicative of HPV-related cytological lesions. When suspicious cases of HPV-related alterations were grouped with positive cases, they showed on average: sensitivity of 25.5%, specificity of 84.4% and positive predictive value (PPV) of 26.8%. When suspicious cases were grouped with negative cases, sensitivity decreased, whereas specificity and PPV increased, as expected. In the second reading, which followed a "guide-list", a decrease in sensitivity was observed, contrasting with a sharp increase of positive predictive value. Among the 16 cytomorphological criteria tested, "koilocytosis", "mild koilocytosis" and "condylomatous parabasal cells" yielded the best predictive value for HPV DNA detection by PCR. In conclusion, despite the low sensitivity, cytopathologic assessment of cervico-vaginal smears leads to a highly specific diagnosis of HPV infection in menopausal women, with PPV of 91.0% when directed by a guide-list of well-defined morphologic criteria.

Biologia da polinização e sistema reprodutivo de Passiflora coccinea Aubl. Em Manaus, Amazonas, Brasil1

Foi estudada a biologia da polinização e o sistema reprodutivo de Passiflora coccinea uma Passifloraceae comum na região Amazônica, conhecida popularmente por maracujá-poranga ou tomé-assu. Este estudo foi realizado em uma área perturbada, na Estação Experimental de Silvicultura Tropical do INPA (03º06'08" S, 59º58'54" W), em agosto e setembro de 1999. Dados de morfometria, horário de abertura e tempo de vida das flores foram obtidos. Diariamente foram observados os animais visitantes, o tipo de alimento procurado, horário e frequência das visitas às flores. Foram realizadas experiências sobre o sistema de reprodução, como também, observada a produção natural de frutos na área. A flor de P. coccinea apresenta a síndrome da ornitofilia pois tem antese diurna, é vermelha, não possui odor e apresenta néctar em abundância. Os beija-flores Amazilia sp. e Phaethornis superciliosus foram considerados seus polinizadores. Duas espécies de Hymenoptera c duas de Lepidoptera foram consideradas "ladras de pólen" e "ladras de néctar", respectivamente, pois não contactam a superfície estigmatífera. O sistema de reprodução desta espécie de maracujá é a xenogamia visto que não foram produzidos frutos por apomixia e por autogamia. A taxa natural de produção de frutos aumentou de 15% para aproximadamente 54% com a realização de polinizações manuais.

Estudo farmacognóstico e atividade in vitro sobre a coagulação sanguínea e agregação plaquetária das folhas de Passiflora nitida Kunth (Passifloraceae)

O gênero Passiflora (Passifloraceae) é utilizado principalmente para tratar doenças do SNC e cardiovasculares. A espécie Passiflora nitida Kunth é comumente conhecida como “maracujá-do-mato". A literatura relata o consumo in natura dos frutos desta espécie pela população local para distúrbios gastrointestinais. Considerando o potencial farmacológico do gênero, este trabalho teve por objetivo realizar estudo de caracterização fitoquímica desta espécie e estudar os efeitos dos extratos aquoso (EA), etanólico (EE) e hexânico (EH) de suas folhas sobre a coagulação sanguínea e agregação plaquetária. Para a caracterização fitoquímica foram realizados testes de cromatografia em camada delgada e ressonância magnética nuclear. O efeito dos extratos sobre a coagulação foi avaliado pelos testes de tempo de protrombina (TP) e tempo de tromboplastina parcial ativada (TTPa). O efeito sobre a agregação plaquetária foi avaliado em plasma rico em plaquetas por método espectrofotométrico, usando adenosina difosfato (ADP) e adrenalina (ADR) como indutores da agregação. Os extratos EA, EE e EH apresentaram atividade coagulante pelo teste do TP e o EE apresentou atividade anticoagulante para o TTPa. Quando induzidos por ADP, os extratos EA, EE e EH apresentaram valores de concentração inibitória 50% (CI50, µg/mL) de 450,5 ± 50,7; 511,2 ± 35,5 e 394,4 ± 8,9, respectivamente, e quando induzidos por ADR apresentaram valores de 438,7 ± 5,2; 21,0 ± 1,9 e 546,9 ± 49,9, respectivamente. O EE apresentou atividade inibitória sobre a agregação. A caracterização fitoquímica foi sugestiva da presença de flavonóides e cumarinas, aos quais podem ser atribuídos, em parte, os efeitos biológicos estudados.

Passiflora fissurosa, uma nova espécie de Passifloraceae para o Amazonas, Brasil

Passiflora fissurosa M.A.D.Souza sp. nov., até o momento conhecida apenas da Reserva Florestal Adolpho Ducke, no Amazonas, Brasil, é descrita e ilustrada. Foi inserida no Subgênero Passiflora, Superseção Passiflora, Seção Laurifoliae e Série Laurifoliae, por apresentar folhas simples e inteiras, pecíolo biglandular, brácteas foliáceas, livres, maiores que 1 cm, pertencendo ao grupo de espécies com as duas séries externas da corona subiguais. Morfologicamente é semelhante a P. nitida, que difere pelo opérculo horizontal-encurvado com margem ereto-divergente, anel nectarífero presente, límen vertical e ovário glabro. A designação do epíteto deve-se à característica do ritidoma, suberoso e profundamente fissurado, característica somente encontrada em P. phellos, também pertencente à Série Laurifoliae, mas do grupo de espécies com a primeira série da corona menor que a segunda.

The role of liquid-based cytology and ancillary techniques in pleural and pericardic effusions: An institutional experience

BACKGROUND: Fine-needle aspiration cytology (FNAC) of serous membrane effusions may fulfil a challenging role in the diagnostic analysis of both primary and metastatic disease. From this perspective, liquid-based cytology (LBC) represents a feasible and reliable method for empowering the performance of ancillary techniques (ie, immunocytochemistry and molecular testing) with high diagnostic accuracy. METHODS: In total, 3171 LBC pleural and pericardic effusions were appraised between January 2000 and December 2013. They were classified as negative for malignancy (NM), suspicious for malignancy (SM), or positive for malignancy (PM). RESULTS: The cytologic diagnoses included 2721 NM effusions (2505 pleural and 216 pericardic), 104 SM effusions (93 pleural and 11 pericardic), and 346 PM effusions (321 pleural and 25 pericardic). The malignant pleural series included 76 unknown malignancies (36 SM and 40 PM effusions), 174 metastatic lesions (85 SM and 89 PM effusions), 14 lymphomas (3 SM and 11 PM effusions), 16 mesotheliomas (5 SM and 11 SM effusions), and 3 myelomas (all SM effusions). The malignant pericardic category included 20 unknown malignancies (5 SM and 15 PM effusions), 15 metastatic lesions (1 SM and 14 PM effusions), and 1 lymphoma (1 PM effusion). There were 411 conclusive immunocytochemical analyses and 47 molecular analyses, and the authors documented 88% sensitivity, 100% specificity, 98% diagnostic accuracy, 98% negative predictive value, and 100% positive predictive value for FNAC. CONCLUSIONS: FNAC represents a primary diagnostic tool for effusions and a reliable approach with which to determine the correct follow-up. Furthermore, LBC is useful for ancillary techniques, such as immunocytochemistry and molecular analysis, with feasible diagnostic and predictive utility.