The inhibitor of the redox activity of APE1/REF-1, APX2009, reduces the malignant phenotype of breast cancer cells.

Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/REF-1) is a multifunctional protein acting on cellular signaling pathways, including DNA repair and redox activities. APE1/REF-1 has emerged as a target for cancer therapy, and its role in breast cancer models would reveal new strategies for cancer therapy. APX2009 is a specific APE1/REF-1 redox inhibitor whose anticancer properties have not been described in breast cancer cells. Here, we investigated the effect of the APX2009 treatment in the breast cancer cell lines MDA-MB-231 and MCF-7. Breast cancer cell lines were cultured, and WST1 and colony formation assays were performed to evaluate cell proliferation. Annexin V-FITC/7-AAD and LDH-Glo™ assays were performed to evaluate cell death. The wound healing assay and Matrigel transwell assay were performed after APX2009 treatment to evaluate the cellular migration and invasion processes, respectively. Our findings demonstrated that APX2009 treatment decreased breast cancer cell proliferative, migratory, and invasive properties. Furthermore, it induced apoptosis in both cell lines. Our study is the first to show the effects of APX2009 treatment on apoptosis in a breast cancer cell. Therefore, this study suggested that APX2009 treatment is a promising anticancer molecule for breast cancer.

Microencapsulation of Renealmia alpinia (Rottb.) Maas pulp pigment and antioxidant compounds by spray-drying and its incorporation in yogurt.

Renealmia alpinia (Rottb.) Maas pulp was processed by spray drying using Maltodextrin (MDX), and Gum Arabic (GA), and the mixture of both encapsulating agents (MDX-GA). Yield, moisture, water activity (a w ), apparent and bulk densities, size and morphology of capsules, color, and antioxidant potential (antioxidant activity, total carotenoids, and phenolic compounds) were analyzed. The encapsulates were incorporated as pigments in yogurt and the stability of antioxidant compounds (1, 7, 14, 21, and 28 days of storage) and the sensory properties were evaluated. The yields of all formulations (MDX, GA, MDX-GA) were around 17.86% with low moisture and a w range values (2.62-3.29% and 0.276-0.309, respectively). The microcapsules presented multiples particle sizes (0.67-27.13 µm) with irregular and smooth surfaces. Furthermore, these capsulates preserved yellow color and the retention of carotenoids was significantly higher with MDX (34.12 mg/100 g of powder), while the phenolic compounds and antioxidant activity increased with GA (474.17 mg GAE/100 g and 552.63 mg TE/100 g of powder, respectively). The main compounds ß-carotene and gallic acid were identified and quantified in positive and negative mode respectively using LC-MS/MS. Finally, the addition of the encapsulated pigments to yogurt allowed to obtain a yellow coloration and the yogurt added with MDX-GA presented the best formulation with not significant changes in antioxidant activity and acceptable sensory attributes up 28 days of storage.

Larvicidal activity of the skin secretion of Rhinella marina and Rhaebo guttatus (Anura: Bufonidae) against the Brazilian malaria vector, Anopheles darlingi (Diptera: Culicidae)

@#Malaria, a mosquito-borne disease, is caused by protozoa of the genus Plasmodium and constitutes a serious public health problem. Because current insecticides used to control malaria face resistance due to continuous use, new alternatives are prompted. Considering this context, and the insecticidal potential of vertebrate venoms/secretions, crude and methanolic extracts from two frog species were tested as larvicides against Anopheles darlingi. Skin secretions of Rhinella marina and Rhaebo guttatus were obtained by manual stimulation. Then, methanol was added to obtain steroidal fractions from both venoms. Mosquitos were captured in suburban areas of Porto Velho and An. darlingi females were later fed with blood and stimulated to oviposit. The larvae were fed with fish food until the 3rd and 4th instars. For the larvicidal assays, crude secretions and methanolic fractions of both frog species were evaluated, and larvae mortality was recorded after 48 hours. Crude extracts and steroidal fractions from both species had larvicidal effects, with an LC50 of 127.5 and 133 ppm for the crude extract and steroidal fraction of R. marina, and an LC50 of 37.5 and 35.8 ppm for the crude extract and steroidal secretion of R. guttatus, respectively. The present work reports for the first time the larvicidal effects of the skin secretions from bufonid species occurring in the western Amazon region. Further studies should be carried out to investigate the purified components responsible for the observed activity.

Evaluation of attractive toxic sugar baits (ATSB) against Aedes aegypti (Diptera: Culicidae) in laboratory.

Toxic baits, widely used against insect pests, are being successfully used to control mosquito vectors. In the present study, basic aspects for Attractive Toxic Sugar Baits (ATSBs) use as a control tool against Aedes aegypti including insecticide dosage, bait composition and plant application under laboratory conditions were evaluated. The Lethal Concentrations (LC 50 and 90) of boric acid (insecticide) Ae. aegypti engorgement and mortality were determined using ATSBs prepared using fruits (guava, mango and cupuaçu) and offered to mosquitoes on cotton discs and also sprayed on a Kalanchoe blossfeldiana plant. LCs of Ae. aegypti males and females did not differ significantly and varied from 0.53 to 2.46%, decreasing from 24 to 48 hours. No significant difference in the proportion of engorged male mosquitoes in ATSB (0.60) and Attractive Sugar Bait (ASB) (0.65) was found, but females engorged more on ASB (control bait) (0.80) compared to ATSBs (0.67). General mortality rate of mosquitoes in ATSB and ASB were 0.81 and 0.10 for males, respectively; 0.61 and 0.12 for females, respectively. Fruit composition affected neither engorgement nor mortality. ATSB applied on plants caused the mortality of males and females ranging from 0.75-0.87 while mortality on ASB sprayed plants varied from 0.07-0.14. Different common fruit juices and a low toxic oral insecticide are readily accepted, engorged and causes a high mortality both males and females Ae. aegypti using ATSBs. Moreover, the use of a common indoor plant in the region sprayed with ATSB under laboratory conditions leads to significant mosquito mortality.

Evaluation of attractive toxic sugar baits (ATSB) against Aedes aegypti (Diptera: Culicidae) in laboratory

@#Toxic baits, widely used against insect pests, are being successfully used to control mosquito vectors. In the present study, basic aspects for Attractive Toxic Sugar Baits (ATSBs) use as a control tool against Aedes aegypti including insecticide dosage, bait composition and plant application under laboratory conditions were evaluated. The Lethal Concentrations (LC 50 and 90) of boric acid (insecticide) Ae. aegypti engorgement and mortality were determined using ATSBs prepared using fruits (guava, mango and cupuaçu) and offered to mosquitoes on cotton discs and also sprayed on a Kalanchoe blossfeldiana plant. LCs of Ae. aegypti males and females did not differ significantly and varied from 0.53 to 2.46%, decreasing from 24 to 48 hours. No significant difference in the proportion of engorged male mosquitoes in ATSB (0.60) and Attractive Sugar Bait (ASB) (0.65) was found, but females engorged more on ASB (control bait) (0.80) compared to ATSBs (0.67). General mortality rate of mosquitoes in ATSB and ASB were 0.81 and 0.10 for males, respectively; 0.61 and 0.12 for females, respectively. Fruit composition affected neither engorgement nor mortality. ATSB applied on plants caused the mortality of males and females ranging from 0.75-0.87 while mortality on ASB sprayed plants varied from 0.07-0.14. Different common fruit juices and a low toxic oral insecticide are readily accepted, engorged and causes a high mortality both males and females Ae. aegypti using ATSBs. Moreover, the use of a common indoor plant in the region sprayed with ATSB under laboratory conditions leads to significant mosquito mortality.

Nasal airway evaluation in obstructive sleep apnoea patients: volumetric tomography and endoscopic findings.

Obstructive sleep apnoea (OSA) results from the recurrent collapse of the upper airway during sleep. Nasal abnormalities influence the stability of the pharynx. The aim of this study was to evaluate the volumetric and anatomical changes of the nasal cavity in patients with OSA. The Nasal Obstruction Symptom Evaluation (NOSE) scale was used to grade nasal obstruction. Sleep-related breathing disorders were evaluated by polysomnography. The nasal airway volume was obtained from computed tomography scans through volumetric reconstruction of the nasal airway. Alterations to the nasal anatomy were identified by nasal fibre-optic endoscopy. Ninety-four patient charts were analyzed. The final sample comprised 32 patients with severe OSA, 16 with moderate OSA, 23 with mild OSA, and 20 without OSA. Three groups were established based on nasal obstruction and OSA. The groups were compared for nasal airway volume (P=0.464) and body mass index (P=0.001). The presence of nasal septum deviation and inferior turbinate hypertrophy were related to the NOSE score (P=0.05 for both), apnoea-hypopnoea index (P=0.03 and P=0.05, respectively), and nasal airway volume (P=0.71 and P=0.78, respectively). In this nasal airway evaluation of OSA patients, the presence of sites of obstruction was correlated with the severity of OSA; this was not the case for the evaluation of the nasal airway volume dimensions.

O papel terapêutico do Programa Farmácia Viva e das plantas medicinais / The therapeutic role of the Program Farmacia Viva and the medicinal plants in the center-south of Piauí

RESUMOEste trabalho realizou um levantamento sobre o uso de plantas medicinais na cidade de Picos-PI, identificou as plantas cultivadas no horto pertencente ao Laboratório Fitoterápico de Picos (LAFIPI), e analisou o uso de fitoterápicos dispensados pelo Programa Farmácia Viva no triênio 2008-2010. Do total dos 750 entrevistados, 37,6% foram homens e 62,4 % mulheres, dentre os quais a maioria não concluiu o segundo grau (69,2%) e 77,2% possuíam renda mensal de até dois salários mínimos. Com relação ao consumo de plantas medicinais, 76,3% afirmaram utilizá-las para tratar doenças, principalmente por considerá-las mais saudáveis (84,8%). A indicação do uso foi orientada, sobretudo, por familiares (82,2%), embora a maioria adquira as plantas em feiras livres (32,8%). Das 127 plantas relatadas, as mais citadas foram erva-cidreira, boldo e hortelã, sendo as folhas a parte mais utilizada (42,3%), predominantemente por infusão (39,4%). As aplicações mais lembradas foram para tratar dores em geral (17%), distúrbios respiratórios (16,5%) e digestivos (16%). As espécies mais cultivadas no horto são chambá (Justicia pectoralis), alecrim pimenta (Lippia sidoides), malva santa (Plectranthus barbatus) e erva cidreira (Lippia alba). O lambedor de chambá foi o fitoterápico mais procurado pela população entre 2008 e 2010. Esse estudo descreveu, pela primeira vez, o uso tradicional de plantas medicinais no município de Picos e demonstrou, também de forma inédita, a relevância de investimentos do Programa Farmácia Viva no município de Picos e sua inclusão no Programa Saúde da Família como forma de disponibilizar à população de baixa renda fitoterápicos produzidos localmente a custos reduzidos.

ABSTRACTThis study performed a research about the use of medicinal plants in Picos city, identifying the plants grown in the garden belonging to the Laboratory of Phytotherapics in Picos (LAFIPI) and analying the use of phytotherapics distributed by the Farmácia Viva Project between 2008-2010. From the total of 750 interviewed participants, 37.6% were men and 62.4 % women. Most of them do not have secondary education (69.2%) and 77.2% had an income of up until two minimum wages. Regarding consumption of medicinal plants, 76.3 % used them to treat diseases, mainly because they considered them to be healthier (84.8%). The indication of use of the plants was mainly suggested by relatives (82.2%), even though the majority of the participants acquires plants in open markets (32.8%). From the 127 plants mentioned, the most cited ones were balm, boldo and mint, and leaves were the most utilized parts (42.3%), predominantly by infusion (39.4%). The most common reasons for the usewere to treat pain in general (17%), and respiratory (16.5%) and digestive disorders (16%). The most cultivated species in the garden were chambá (Justicia pectoralis), alecrim pimenta (Lippia sidoides), malva santa (Plectranthus barbatus) and erva cidreira (Lippia alba). The "chambá licker" was the most herbal medicine searched by the population between 2008 and 2010. This investigation described, for the first time, the traditional use of medicinal plants in Picos and demonstrated,alsounprecedentedly, the relevance of investments in the Farmácia Viva Project in Picos city and its inclusion in the Family Health Program as a strategyin order to provide locally producedphytotherapics for low-income population at low costs.

Dung beetles (Coleoptera: Scarabaeoidea) in three landscapes in Mato Grosso do Sul, Brazil.

Dung beetles (Coleoptera: Scarabaeoidea) in three landscapes in Mato Grosso do Sul, Brazil. Dung Beetles are important for biological control of intestinal worms and dipterans of economic importance to cattle, because they feed and breed in dung, killing parasites inside it. They are also very useful as bioindicators of species diversity in agricultural or natural environments. The aims of this paper were to study the species richness, and abundance of dung beetles, helping to answer the question: are there differences in the patterns of dung beetle diversity in three environments (pasture, agriculture and forest) in the municipality of Dourados, in the state of Mato Grosso do Sul. A total of 105 samplings were carried out weekly, from November 2005 to November 2007, using three pitfall traps in each environment. The traps were baited with fresh bovine dung, and 44,355 adult dung beetles from 54 species were captured: two from Hyborosidae and 52 from Scarabaeidae. Five species were constant, very abundant and dominant on the pasture, two in the agricultural environment, and two in the environment of Semideciduous forest. Most of the species were characterised as accessories, common and not-dominant. The species with higher abundance was Ataenius platensis Blanchard, 1844. The indexes of Shannon-Wiener diversity were: 2.90 in the pasture, 2.84 in the agricultural environment and 2.66 in the area of native forest. The medium positive presence of dung beetles in the traps in each environment were: 36.88, 42.73 and 20.18 individuals per trap, in the pasture, agricultural environment and in the native forest, respectively. The pasture environment presented a higher diversity index. The species diversity of dung beetles was superior where there was higher abundance and regularity of resource (bovine dung).

In vivo assessment of specific cytotoxic T lymphocyte killing.

The direct killing of target cells by cytotoxic T lymphocytes (CTLs) plays a fundamental role in protective immunity to viral, bacterial, protozoan and fungi infections, as well as to tumor cells. In vivo cytotoxic assays take into account the interaction of target and effector cells in the context of the proper microenvironment making the analysis biologically more relevant than in vitro cytotoxic assays. Thus, the development, improvement and validation of in vivo methods are necessary in view of the importance of the results they may provide. We describe and discuss in this manuscript a method to evaluate in vivo specific cytotoxic T lymphocyte killing. We used as model system mice immunized with human recombinant replication-deficient adenovirus 5 (HAd5) containing different transgenes as the trigger of a CTL-mediated immune response. To these mice, we adoptively transferred syngeneic cells labeled with different vital fluorescent dyes. Donor cells were pulsed (target) or not (control non-target) with distinct CD8 T-cell epitopes, mixed in a 1:1 ratio and injected i.v. into immunized or non-immunized recipient mice. After 18-24h, spleen cells are collected and analysed by flow cytometry. A deviation from the 1:1 ratio of control and target cell populations indicates antigen specific lysis of target cells.

Role of adhesion molecules and proliferation hyperplasic, pre neoplastic and neoplastic lesions in canine prostate.

E-cadherin and beta-catenin are component of adherens junctions in epithelial cells. Loss of these proteins have been associated with progression of prostatic diseases. We performed immunohistochemistry for E-cadherin, beta-catenin and Ki-67 on canine prostatic lesions. We analyzed the expression of these antibodies in benign prostatic hyperplasia (BPH, n = 22), in pre neoplastic lesions Prostatic Intra-epithelial Neoplasia (PIN), n = 3 and Prostatic Inflammatory Atrophy (PIA), n = 7 and prostate carcinoma (PC, n = 10). In this study, a membranous expression of E-cadherin and beta-catenin and nuclear expression of Ki-67 antigen were demonstrated. The proliferative index was statistically different between carcinomas and BPH and carcinomas and pre-neoplastic lesions. Like in men, the reduction of E-cadherin and increase of Ki-67 expression in neoplastic lesions in dog prostate may be related to the carcinogenic process in this gland.

Óxido nítrico, GSTP-1 e p53: qual o papel desses biomarcadores nas lesões prostáticas do cão? / Nitric oxide, GSTP-1 and p53: what's the role of these biomarkers in dog´s prostate lesions?

Confeccionou-se um microarranjo de tecido (TMA) com 146 amostras de lesões prostáticas caninas. Este continha 17,2 por cento de hiperplasia prostática benigna (HPB), 32,4 por cento de atrofia inflamatória proliferativa (PIA), 2,6 por cento de prostatite, 8,6 por cento de focos de neoplasia intraepitelial prostática (PIN), 29,1 por cento de carcinomas e 9,3 por cento de próstatas normais. Cortes histológicos sequenciais foram feitos e utilizados para reação de imunoistoquímica com os anticorpos primários anti-p-53, anti-NOS-2 e anti-GSTP. Avaliou-se de cada core o escore de células marcadas para cada anticorpo utilizado. Os resultados foram tabulados por grupo diagnóstico e submetidos ao teste Tuckey. Os carcinomas prostáticos do cão e a PIA apresentaram maior número de amostras (41) com mais de 75 por cento das células positivas para NOS-2, demonstrando a influência do estresse oxidativo no desenvolvimento dessas lesões. As próstatas normais e as afecções desta glândula, HPB, PIA, PIN, prostatite e carcinoma, expressaram a proteína GSTP-1, o que conferiu proteção ao tecido prostático canino a danos oxidativos. A proteína p53 estava presente em todas as amostras estudadas, incluindo o tecido prostático normal, porém as lesões prostáticas apresentaram maior número de amostras com escores mais elevados de marcação (escores três e quatro), presente em 95 por cento dos focos de PIA e carcinoma. Concluiu-se que o aumento de expressão de óxido nítrico nas lesões prostáticas no cão e a expressão de GSTP-1 podem ter protegido o tecido prostático canino e que a expressão de p53 foi positiva e uniforme nas próstatas normais e com lesões hiperplásicas e displásicas.

A tissue microarray (TMA) with 149 samples of canine prostatic lesions contained 17.2 percent benign prostatic hyperplasia (BPH), 32.4 percent proliferative inflammatory atrophy (PIA), 2.6 percent prostatitis, 8.6 percent foci prostatic intraepithelial neoplasia (PIN), 29.1 percent carcinomas and 9.3 percent normal prostates. Sequential histological sections were made and used for immunohistochemistry reaction with primary antibodies anti p-53, anti-NOS-2 and anti-GSTP. The score for each antibody employed was evaluated. The results were tabulated by diagnostic group and subjected to Tuckey test. Prostatic carcinomas and PIA had a higher number of samples (41) with over 75 percent of cells positive for NOS-2, demonstrating the influence of oxidative stress in the development of these lesions. The prostates of normal dogs, as well as the disorders of this gland (BPH, PIA, PIN, prostatitis and carcinoma), expressed GSTP-1 protein, which gives protection to the canine prostate tissue against oxidative damage. The p53 protein was present in all samples studied, including normal prostate tissue, but the prostatic lesions had a higher number of samples with higher scores (more than 50 percent of positive cells), in 95 percent of foci of PIA and carcinoma. It was found that an increased expression of NO in prostatic lesions of dogs and that the expression of GSTP-1 can protect the canine prostate tissue, which would contribute to the low frequency of prostate adenocarcinoma in this species. The expression of p53 was positive in all lesions as well as the in normal prostate.

Uso de células mononucleares da medula óssea no tratamento de tendinites induzidas experimentalmente em equinos / The use of mononuclear cells from bone marrow on the treatment of experimentally induced tendinitis in horses

Foram induzidas lesões no tendão flexor digital superficial (TFDS) de ambos os membros anteriores de seis equinos, seguidas de implante autólogo de células da fração mononuclear de medula óssea em apenas um membro de cada animal. Os animais foram avaliados por parâmetros clínicos, ultrassonográficos, histopatológico e imunoistoquímico. Paralelamente, realizou-se o cultivo de novas amostras para a caracterização das células utilizando-se marcadores CD34 e CD45 por meio da citometria de fluxo, confirmando a presença de células mesenquimais na fração mononuclear. A caracterização das fibras colágenas tipo I e tipo III no tecido neoformado mostrou melhora na qualidade da cicatrização tendínea dos membros tratados. A terapia com implante autólogo das células da fração mononuclear melhorou a organização tecidual e a sua qualidade, apresentando maior expressão significativa para colágeno tipo I.

The present study was developed inducing a lesion in the SDFT of both thoracic limbs of six horses followed by autologous implantation of mononuclear cells from bone marrow in only one affected limb of each horse. The horses were evaluated through clinical and ultrasonography exams, and through histopathology and immunohistochemistry patterns. Concomitantly, new samples were cultivated and characterized using CD34 and CD45 markers, proving the presence of mesenchymal cells in the mononuclear fraction. The characterization of collagen fibers type I and type III in the new tissue has showed an improvement in tendon healing in treated limbs. The therapy with autologous implant of the mononuclear fraction has improved tissue organization and its quality, having a significanlyt higher expression of collagen type I.

Detecção imunoistoquímica de receptores de estrógeno e progesterona no endométrio de vacas Nelore (Bos taurus indicus) durante o anestro pós-parto / Immunohistochemical detection of receptors for estrogen and progesterone in endometrial glands and stroma during the pospartum anestrus in Nelore (Bos taurus indicus) cows

Foram utilizadas 24 vacas Nelore P.O., em anestro pós-parto, diagnosticado pelo histórico reprodutivo, por avaliações ultrassonográficas transretais e por dosagem de progesterona plasmática, que foram submetidas à colheita de fragmento uterino via transcervical. Os animais foram divididos em dois grupos conforme o máximo diâmetro folicular: grupo 1: folículos <6mm (n=12); grupo 2: folículos >6mm (n=12). Para avaliar receptor de estrógeno e receptor de progesterona no epitélio glandular e no estroma, foi utilizada a técnica de imunoistoquímica. Altas contagens relativas e alta intensidade de marcação para receptor de estrógeno e progesterona no epitélio glandular e no estroma foram observadas nos dois grupos. No entanto, a intensidade de marcação para o receptor de progesterona no epitélio glandular foi mais alta no grupo 2 comparado ao grupo 1. Quando o epitélio glandular e o estroma foram comparados, o número relativo de receptor de estrógeno no grupo 1 foi mais alto no epitélio glandular comparado ao estroma, e a intensidade de marcação para o receptor de progesterona no grupo 2 foi mais alta no epitélio glandular comparado ao estroma. Os resultados sugerem que os mecanismos que controlam a expressão de receptores no anestro são semelhantes aos observados durante o ciclo estral.

Twenty-four postpartum anestrous Nelore purebred cows were used in the study. Anestrous was determined based on the reproductive history which was confirmed in each cow based on plasma progesterone concentration and by transrectal ultrasonography. Endometrial biopsies were collected. The animals were separated into two groups according to maximum follicular diameter- Group 1: follicles <6mm (n=12) and Group 2: follicles >-6mm follicles (n=12). The immunohistochemistry technique was employed to evaluate the presence of estrogen and progesterone receptors in the uterine glandular epithelium and stroma. High counts of positive nuclei and high intensity of immunostain for estrogen and progesterone receptors in the glandular epithelium and stroma were observed in the two groups. However, the immunostain intensity of progesterone receptors in the glandular epithelium was higher in Group 2 compared to Group 1. When glandular epithelium and stroma were compared within each group, the relative number of estrogen receptors in the Group 1 was higher in the glandular epithelium compared to stroma and the immunostain intensity for the progesterone receptor in Group 2 was higher in the glandular epithelium compared to stroma. The results suggest that the mechanisms that control the expression of endomerial receptors in the anestrus are similar to the ones observed during the estrus cycle.

Hot and cold water infusion aroma profiles of Hibiscus sabdariffa: fresh compared with dried.

UNLABELLED: Calyxes from the Roselle plant (Hibiscus sabdariffa L.) were used to prepare cold (22 °C for 4 h) and hot (98 °C for 16 min) infusions/teas from both fresh and dried forms. Aroma volatiles were extracted using static headspace SPME and analyzed using GC-MS and GC-O with 2 different columns (DB-5 and DB-Wax). Totals of 28, 25, 17, and 16 volatiles were identified using GC-MS in the dried hot extract (DHE), dried cold extract (DCE), fresh hot extract (FHE), and fresh cold extract (FCE) samples, respectively. In terms of total GC-MS peak areas DHE ≫ DCE > FHE ≫ FCE. Nonanal, decanal, octanal, and 1-octen-3-ol were among the major volatiles in all 4 beverage types. Thirteen volatiles were common to all 4 teas. Furfural and 5-methyl furfural were detected only in dried hibiscus beverages whereas linalool and 2-ethyl-1-hexanol were detected only in beverages from fresh hibiscus. In terms of aroma active volatiles, 17, 16, 13, and 10 aroma active volatiles were detected for DHE, DCE, FHE, and FCE samples, respectively. The most intense aroma volatiles were 1-octen-3-one and nonanal with a group of 4 aldehydes and 3 ketones common to all samples. Dried samples contained dramatically higher levels of lipid oxidation products such as hexanal, nonanal, and decanal. In fresh hibiscus extracts, linalool (floral, citrus) and octanal (lemon, citrus) were among the highest intensity aroma compounds but linalool was not detected in any of the dried hibiscus extracts. PRACTICAL APPLICATION: Hibiscus teas/infusions are one of the highest volume specialty botanical products in international commerce. The beverage is consumed for both sensory pleasure and health attributes and is prepared a number of ways throughout the world. Although color and taste attributes have been examined, little information is known about its aroma volatiles and no other study has compared extractions from both fresh and dried as well as extraction temperature differences. This is also, apparently, the first study to identify the aroma active volatiles in hibiscus beverages using GC-olfactometry. Manufacturers and consumers will now have a better understanding of why hibiscus teas prepared in different ways from either fresh or dried forms have a different flavor quality and intensity.

Lavado traqueobrônquico por via nasotraqueal como metodologia de colheita de células do trato respiratório de ovinos sadios e portadores de afecções pulmonares / Tracheobronchial lavage by nasotracheal via to collect cells of respiratory tract of health and broncopneumonic ovines

Os estudos das secreções traqueobrônquicas são amplamente utilizados nas pesquisas de doenças pulmonares nas diversas espécies animais, inclusive no homem. Os objetivos desta pesquisa foram a viabilização da técnica de colheita de lavado traqueobrônquico na espécie ovina e o estudo da relação clínico-citológica do lavado de ovinos portadores de broncopneumonia e sadios. Foram utilizados 33 ovinos, 18 sadios e 15 portadores de enfermidade respiratória com sinais clínicos de envolvimento das vias aéreas, divididos nos respectivos grupos, GS e GD. Após o exame físico foi realizado o lavado traqueobrônquico por via nasotraqueal. A colheita do lavado foi feita com a inoculação e aspiração de solução fisiológica estéril. As amostras foram processadas citologicamente através de citocentrifugação e coradas pelos métodos Wright-Giemsa e Shorr. Tanto a contagem total de células epiteliais quanto o número de hemácias por mililitro foi maior no grupo de animais com broncopneumonia. Nos animais sadios notou-se predomínio de macrófagos, seguido por células epiteliais cilíndricas, neutrófilos e linfócitos. No grupo de animais doentes havia menor número de macrófagos, e predomínio da população de neutrófilos. Por ser de fácil realização, pouco dispendiosa e pela obtenção representativa de material, a técnica estudada mostrou-se eficaz na obtenção de fluidos traqueobrônquicos e, portanto um bom método de colheita de células para uso nas pesquisas de vias aéreas.

Exams of tracheobronchial secretions are widely used in research of lung diseases in several animal species, including men. The objective of this paper was to investigate feasibility of the technique for harvesting trachea-bronchial lavage in sheep and to verify its clinic-cytological relationship in healthy sheep and others suffering from pneumonia. In this study 33 sheep were used, 18 healthy and 15 sheep with clinical signs of respiratory disease, divided into groups GS and GD. Clinic examination was performed by obtaining tracheobronchial lavage through the nasal route. The crop was washed with the addition and aspiration of sterile saline. The samples were processed by cytospin and staining with Wright-Giemsa and Shorr. The total cell count and the number of red blood cells per milliliter was higher in the group of sheep with bronchopneumonia. In healthy animals predominance of macrophages was noted, followed by the number of epithelial cells, neutrophils and lymphocytes. In the group of sick animals fewer macrophages and predominance of the neutrophil population were noted. Because of its easy performance to obtain representative material, the technique studied was considered effective to obtain tracheobronchial fluid and therefore is a good method to harvest cells for research of the airways.

Frequency of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma species in cervical samples.

We investigated the relative frequencies of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma sp. in cervical samples. PCR analyses were performed in ectocervical and endocervical samples from 224 patients attending public health services in Belo Horizonte and Contagem, Minas Gerais Brazil. A high prevalence of colonisation of the cervix (6.3% for C. trachomatis, 4.0% for N. gonorrhoeae, 0.9% for M. genitalium, 21.9% for M. hominis, 38.4% for Ureaplasma sp.) was demonstrated not only for pathogens classically associated to cervicitis (C. trachomatis and N. gonorrhoeae), but also for M. hominis and Ureaplasma sp. These findings may be useful to guide more adequate diagnosis to interrupt transmission and to avoid negative impacts on the female reproductive tract.

Plasma concentrations and placental immunostaining of interleukin-10 and tumornecrosis factor-α as predictors of alterations in the embryo-fetal organism and the placental development of diabetic rats

Interleukin-10 (IL-10) appears to be the key cytokine for the maintenance of pregnancy and inhibits the secretion of inflammatory cytokines such as tumor necrosis factor-α (TNF-α). However, there are no studies evaluating the profile of these cytokines in diabetic rat models. Thus, our aim was to analyze IL-10 and TNF-α immunostaining in placental tissue and their respective concentrations in maternal plasma during pregnancy in diabetic rats in order to determine whether these cytokines can be used as predictors of alterations in the embryo-fetal organism and in placental development. These parameters were evaluated in non-diabetic (control; N = 15) and Wistar rats with streptozotocin (STZ)-induced diabetes (N = 15). At term, the dams (100 days of life) were killed under anesthesia and plasma and placental samples were collected for IL-10 and TNF-α determinations by ELISA and immunohistochemistry, respectively. The reproductive performance was analyzed. Plasma IL-10 concentrations were reduced in STZ rats compared to controls (7.6 ± 4.5 vs 20.9 ± 8.1 pg/mL). The placental scores of immunostaining intensity did not differ between groups (P > 0.05). Prevalence analysis showed that the IL-10 expression followed TNF-α expression, showing a balance between them. STZ rats also presented impaired reproductive performance and reduced plasma IL-10 levels related to damage during early embryonic development. However, the increased placental IL-10 as a compensatory mechanism for the deficit of maternal regulation permitted embryo development. Therefore, the data suggest that IL-10 can be used as a predictor of changes in the embryo-fetal organism and in placental development in pregnant diabetic rats.

Plasma concentrations and placental immunostaining of interleukin-10 and tumor necrosis factor-α as predictors of alterations in the embryo-fetal organism and the placental development of diabetic rats.

Interleukin-10 (IL-10) appears to be the key cytokine for the maintenance of pregnancy and inhibits the secretion of inflammatory cytokines such as tumor necrosis factor-α (TNF-α). However, there are no studies evaluating the profile of these cytokines in diabetic rat models. Thus, our aim was to analyze IL-10 and TNF-α immunostaining in placental tissue and their respective concentrations in maternal plasma during pregnancy in diabetic rats in order to determine whether these cytokines can be used as predictors of alterations in the embryo-fetal organism and in placental development. These parameters were evaluated in non-diabetic (control; N = 15) and Wistar rats with streptozotocin (STZ)-induced diabetes (N = 15). At term, the dams (100 days of life) were killed under anesthesia and plasma and placental samples were collected for IL-10 and TNF-α determinations by ELISA and immunohistochemistry, respectively. The reproductive performance was analyzed. Plasma IL-10 concentrations were reduced in STZ rats compared to controls (7.6 ± 4.5 vs 20.9 ± 8.1 pg/mL). The placental scores of immunostaining intensity did not differ between groups (P > 0.05). Prevalence analysis showed that the IL-10 expression followed TNF-α expression, showing a balance between them. STZ rats also presented impaired reproductive performance and reduced plasma IL-10 levels related to damage during early embryonic development. However, the increased placental IL-10 as a compensatory mechanism for the deficit of maternal regulation permitted embryo development. Therefore, the data suggest that IL-10 can be used as a predictor of changes in the embryo-fetal organism and in placental development in pregnant diabetic rats.

Overexpression of vimentin in canine prostatic carcinoma.

Canine prostatic tumours exhibit similarities to those of man and may represent a useful model system to explore the mechanisms of cancer progression. Tumour progression to malignancy requires a change from an epithelial phenotype to a fibroblastic or mesenchymal phenotype. Vimentin expression is associated with the invasive phenotype of human prostate cancer cells. The aim of the present study was to characterize immunohistochemically the expression of vimentin by canine prostatic carcinomas. Primary carcinomas and metastatic tumour foci both showed vimentin expression. This finding suggests that the acquisition of the epithelial-mesenchymal transition phenotype in canine prostatic carcinoma may be characterized by the presence of mesenchymal intermediate filament (vimentin) that could lead to a higher likelihood of metastasis.

Immunodominance: a new hypothesis to explain parasite escape and host/parasite equilibrium leading to the chronic phase of Chagas' disease?

Intense immune responses are observed during human or experimental infection with the digenetic protozoan parasite Trypanosoma cruzi. The reasons why such immune responses are unable to completely eliminate the parasites are unknown. The survival of the parasite leads to a parasite-host equilibrium found during the chronic phase of chagasic infection in most individuals. Parasite persistence is recognized as the most likely cause of the chagasic chronic pathologies. Therefore, a key question in Chagas' disease is to understand how this equilibrium is established and maintained for a long period. Understanding the basis for this equilibrium may lead to new approaches to interventions that could help millions of individuals at risk for infection or who are already infected with T. cruzi. Here, we propose that the phenomenon of immunodominance may be significant in terms of regulating the host-parasite equilibrium observed in Chagas' disease. T. cruzi infection restricts the repertoire of specific T cells generating, in some cases, an intense immunodominant phenotype and in others causing a dramatic interference in the response to distinct epitopes. This immune response is sufficiently strong to maintain the host alive during the acute phase carrying them to the chronic phase where transmission usually occurs. At the same time, immunodominance interferes with the development of a higher and broader immune response that could be able to completely eliminate the parasite. Based on this, we discuss how we can interfere with or take advantage of immunodominance in order to provide an immunotherapeutic alternative for chagasic individuals.