Liposomes co-encapsulating doxorubicin and glucoevatromonoside derivative induce synergic cytotoxic response against breast cancer cell lines.

Cancer is one of the main causes of death in the world and thus a global public health problem. Among the treatments available for cancer are surgery, radiotherapy, and chemotherapy. Currently, there is increased interest in the combination of two or more antitumor agents to achieve a synergistic effect in cancer therapy. Doxorubicin (DOX), a chemotherapeutic which has a potent antineoplastic action, has been used in the treatment of various tumors. However, the use of DOX is limited, mainly due to the cardiotoxicity. Therefore, nanostructured systems, such as liposomes, have been developed to carry this drug and target the tumor region, since tumor tissues present enhanced permeability and retention for nanosystems. Cardiac glycosides, such as digitoxin, have recently shown great antitumor potential despite the low therapeutic index which may limit their use. Furthermore, some compounds of this class have low water solubility, which makes their in vivo administration difficult. In this context, liposomes represent a valid strategy to carry simultaneously antitumor drugs allowing their intravenous administration. In this study, liposomes loaded with glucoevatromonoside containing peracetylated glucose hydroxyl groups (GEVPG) and DOX at molar ratio of 1:1 (SpHL-GEVPG:DOX 1:1) were developed, and their chemical and physicochemical properties were evaluated. This formulation presented a combination index (CI) lower than 1 at inhibitory concentration of 90 % growth (IC90) for three human breast tumor lines evaluated (0.52 ± 0.39 for MDA-MB-231, 0.19 ± 0.13 for MCF-7, and 0.99 ± 0.09 for SKBR-3). These results indicate a synergistic cytotoxic effect of the GEVPG and DOX combination encapsulated in liposomes. In addition, SpHL-GEVPG:DOX 1:1 presented selectivity towards these cancer cells. Long-term in vitro cytotoxicity studies demonstrated that MDA-MB-231 surviving cells after treatment with SpHL-GEVPG:DOX 1:1 did not recover proliferation capacity after 21 d. From the studies of cell cycle and death pathway evaluation, it was observed that SpHL-GEVPG:DOX 1:1 arrested the cell cycle in the G2/M phase and similarly induced apoptosis and necrosis. However, SpHL-GEVPG:DOX at molar ratio of 1:1 showed lower induction of both apoptotic and necrotic pathways compared to free DOX and SpHL-DOX, suggesting that the mechanism of death involved may not be related to necrosis or apoptosis. Lastly, SpHL-GEVPG:DOX 1:1 showed a good storage stability for 90 d at 4 °C. Therefore, the results of the present work indicate the potential use of SpHL-GEVPG:DOX 1:1 as a new anticancer formulation.

Combined use of chemical and biological compounds to control hookworm.

The aim of this study was to evaluate the combined use of different chemical (albendazole, ivermectin, glycerine and Vaseline) and biological (Monacrosporium thaumasium) compounds in the control of Ancylostoma caninum. Infective larvae of A. caninum were obtained from coprocultures of positive faeces from naturally infected dogs. We used 1% ivermectin, 1% albendazole, 100% glycerine, 100% Vaseline and an isolate of the nematophagous fungus M. thaumasium (NF34), alone or in combinations. Next, an experimental test was set up with 16 groups in microtubes, with a 24-h interaction. The groups (G1 to G15) that contained any chemical or biological compound (NF34) and/or their combined use (chemical + biological) showed a difference in relation to the control group, except G5 - Vaseline 100% without combinations. It was concluded that, even on an experimental basis, the combined use of anthelmintic drugs with biological control was efficient; however, more studies must be carried out in order to elucidate the synergistic action between chemical and biological compounds to be used in the effective control of hookworms in the future.

Systematic review of the biological variation data for diabetes related analytes.

BACKGROUND: Objective interpretation of laboratory test results used to diagnose and monitor diabetes mellitus in part requires the application of biological variation data (BVD). The quality of published BVD has been questioned. The aim of this study was to quality assess publications reporting BVD for diabetes-related analytes using the Biological Variation Data Critical Appraisal Checklist (BIVAC); to assess whether published BVD are fit for purpose and whether the study design and population attributes influence BVD estimates and to undertake a meta-analysis of the BVD from BIVAC-assessed publications. METHODS: Publications reporting data for glucose, HbA1c, adiponectin, C-peptide, fructosamine, insulin like growth factor 1 (IGF-1), insulin like growth factor binding protein 3 (IGFBP-3), insulin, lactate and pyruvate were identified using a systematic literature search. These publications were assessed using the BIVAC, receiving grades A, B, C or D, where A is of highest quality. A meta-analysis of the BVD from the assessed studies utilised weightings based upon BIVAC grades and the width of the data confidence intervals to generate global BVD estimates. RESULTS: BIVAC assessment of 47 publications delivered 1 A, 3 B, 39C and 4 D gradings. Publications relating to adiponectin, C-peptide, IGF-1, IGFBP-3, lactate and pyruvate were all assessed as grade C. Meta-analysis enabled global BV estimates for all analytes except pyruvate, lactate and fructosamine. CONCLUSIONS: This study delivers updated and evidence-based BV estimates for diabetes-related analytes. There remains a need for delivery of new high-quality BV studies for several clinically important analytes.

Long-circulating and fusogenic liposomes loaded with a glucoevatromonoside derivative induce potent antitumor response.

Cancer is an important public health problem, being one of the leading causes of death worldwide. Most antineoplastic agents cause severe toxic effects and some types of cancer do not respond or are resistant to the existing pharmacotherapy, necessitating the research and development of new therapeutic strategies. Cardenolides have shown significant antitumor activity due to their ability to inhibit the Na+K+ATPase enzyme, and the expression of this enzyme is increased in tumor cells. Glucoevatromonoside containing peracetylated glucose hydroxyl groups (GEVPG) is a cardenolide derivative that has low solubility in aqueous media, which constitutes a barrier to its potential biological applications. In this context, the use of liposomes represents a promising strategy to deliver GEVPG, thus allowing its intravenous administration. In this study, long-circulating and fusogenic liposomes containing GEVPG (SpHL-GEVPG) were developed, and their chemical and physicochemical properties were evaluated. SpHL-GEVPG presented adequate properties, including a mean diameter of 182.2 ± 2.7 nm, a polydispersity index equal to 0.36 ± 0.03, a zeta potential of -2.37 ± 0.31 mV, and a GEVPG entrapment of 0.38 ± 0.04 mg/mL. Moreover, this formulation showed a good stability after having been stored for 30 days at 4 °C. The cytotoxic studies against breast (MDA-MB-231, MCF-7, and SKBR-3) and lung (A549) cancer cell lines demonstrated that SpHL-GEVPG treatment significantly reduced the cell viability. In addition, the SpHL-GEVPG formulation presented a good selectivity toward these cancer cells. The evaluation of the therapeutic efficacy of the treatment with SpHL-GEVPG showed a potent anticancer effect in an A549 human lung cancer xenograft model. SpHL-GEVPG administered at doses of 1.0 and 2.0 mg/kg (i.v.) induced antitumor effect comparable to paclitaxel given at dose of 10 mg/kg (i.v.) to mice. Therefore, the results of the present work indicate the potential applicability of SpHL-GEVPG as a new anticancer formulation.

Postpartum follow up of gestational diabetes in a Tertiary Care Center.

BACKGROUND: Gestational diabetes is a risk factor for future development of type 2 diabetes. The primary aim of this study was to estimate the prevalence of postpartum glucose tolerance status evaluation in pregnancies complicated by gestational diabetes 6-12 weeks after delivery. The secondary one was to identify the factors that are implicated with postpartum glucose retesting. METHODS: This was a retrospective study performed with a cohort of women with gestational diabetes, with prenatal care and delivery at a tertiary care center, from January 2013 to April 2017. The diagnosis of gestational diabetes was based on IADPSG criteria (Fasting ≥ 92 mg/dl, 1 h ≥ 180 mg/dl and/or 2 h ≥ 153 mg/dl, respectively) and the diagnosis of type 2 diabetes and prediabetes were made using the 2016 ADA's criteria (fasting and 2 h after glucose load ≥ 126 mg/dl and/or ≥ 200 and 100-125 mg/dl and/or 140 and 199 mg/dl, respectively). All women had an appointment scheduled 6-12 weeks postpartum with the results of a 75-g oral glucose tolerance test (OGTT). RESULTS: Of the 152 evaluated women, 21 (13.8%) returned with the postpartum OGTT results. Of these, 9 (45.0%) had a diagnosis of prediabetes. The use of insulin during gestation was the only factor implicated in a higher adherence rate to postpartum testing OR 6.33 (p 0.002). No significance was found for other demographic and clinical variables (age, family income, years of study, parity, gestational age at first visit, smoking, family history of type 2 diabetes, diagnosis of gestational diabetes before the third trimester, pregestational body mass index, previous history of gestational diabetes and ethnicity). CONCLUSION: The majority of patients with gestational diabetes did not return postpartum to perform OGTT and in our study the only factor implicated in a higher postpartum return was the use of insulin during pregnancy. Considering that 45.0% were diagnosed with prediabetes, diabetes care teams should initially identify non-adherent patients.

Viabilidade da técnica de transposição uretral pré-púbica mediante secção peniana em cadáveres de cães / Viability of the prepubic urethral transposition technique by penile section in dog cadavers

Descreve-se a viabilidade da técnica cirúrgica de transposição uretral pré-púbica em cadáveres de cães machos, adultos, advindos de um hospital veterinário, a fim de se avaliar a manutenção da luz uretral a partir de um desvio uretral pré-púbico, descrevendo-se detalhadamente a abordagem e as estruturas anatômicas envolvidas. A técnica consistiu inicialmente da orquiectomia, seguida de celiotomia retroumbilical, tração vesical cranial, secção transversa da uretra membranosa a 1cm caudal à próstata, espatulação e reparo da borda livre. A seguir, divulsão e secção do pênis 1,5cm caudal ao osso peniano, espatulação da borda uretral peniana e transposição desta em direção à cavidade abdominal, fazendo-se anastomose por meio de sutura interrompida simples à uretra membranosa. Foi mensurado o comprimento uretral desde o meato externo até o início da uretra prostática em dois momentos: pré e pós-transposição. Ao final, avaliou-se, por meio de uretrografia retrógrada de contraste positivo, o diâmetro e o selamento na anastomose uretral e o fluxo do contraste até a bexiga. A transposição uretral pré-púbica é uma técnica cirúrgica exequível, anatomicamente compatível com a espécie canina e capaz de manter o diâmetro uretral e o fluxo do contraste de forma satisfatória, com manutenção estética externa idêntica a um cão macho orquiectomizado.(AU)

We describe the viability of the surgical technique of pre-pubic urethral transposition in cadavers of adult male dogs, coming from a veterinary hospital to evaluate the maintenance of urethral light from a prepubic urethral deviation, describing in detail the approach and anatomical structures involved. The technique consisted initially of orchiectomy, followed by retroumbilical celiotomy, cranial vesical traction, transverse section of the membranous urethra at 1cm caudal to prostate, spatulation and free edge repair. Next, the penis divulsion and section 1.5cm caudal to the penile bone, spreading the penile urethral border and transposing it towards the abdominal cavity, making an anastomosis of simple interrupted suture to the membranous urethra. The dogs had the urethral length measured from the external meatus to the beginning of the prostatic urethra at two moments: pre-transposition and post-transposition. Postoperative retrograde urethrography was used to evaluate the diameter and sealing of the urethral anastomosis and contrast flow to the bladder. In conclusion, the prepubic urethral transposition is a feasible surgical technique, anatomically compatible with the canine species and able to maintain the urethral diameter and contrast flow satisfactorily, with an external esthetic maintenance identical to an orchiectomized male dog.(AU)

Transposição e anastomose uretral pré-púbica em um cão macho com estenose extensa da uretra intrapélvica / Prepubic urethral transposition in a male dog with extensive stenosis of the pelvic urethra

We report the use of a new surgical technique in a male dog affected by extensive stenosis of intrapelvic urethra through a antepubic urethral deviation, as an alternative to prepubic urethrostomy and ablation of the external genitalia. The technique consisted initially of orchiectomy, followed by retroumbilical celiotomy, transverse section of the penis in the pre-scrotal region and transposition of this towards the abdominal cavity by making anastomosis to the prostatic urethra. The dog was evaluated clinically and by urethrography retrograde positive contrast for up to four years after the procedure without any clinical signs, changes in urine stream or stenosis image. It is concluded that the pre-pubic urethral transposition is a viable alternative treatment for this dog affected by extensive stenosis of the membranous urethra.(AU)

Enhanced expression of PD-L1 and IFN-γ on dendritic cells is associated with BCG-induced Th2 inhibition.

Accumulating evidence indicates that the exposure to Mycobacterium bovis bacillus Calmette-Guérin (BCG) prevents the development of allergy and the airway dendritic cells (DCs) may be involved in this protective effect. However, studies to better characterize the specific interactions between BCG and DCs and their role in this mycobacteria-mediated Th2 cell suppression are still ongoing. This study aimed to evaluate the effect of the neonatal BCG vaccination in the innate immune response in a mouse model of ovalbumin (OVA)-induced airway inflammation. BCG treated neonatal BALB/c mice were sensitized and challenged with aerosolized OVA. Twenty-four hours after the last challenge, samples were collected for analysis. The intranasal BCG treatment inhibited the allergic Th2-response by decreasing the allergen-induced eosinophilic inflammation, EPO activity, CCL11, IL-25, TSLP, IL-4 and IL-5 lung levels, and serum levels of IgE. Mycobacteria treatment increased lung levels of IL-10 and TGF-ß, and the TLR2 and TLR4 expressions by pulmonary CD11c+CD103+CD8α+ DCs. Additionally an enhanced expression of PD-L1 was observed besides an increased production of IFN-γ by these cells. These results indicated that neonatal BCG vaccination inhibits key features of allergic airway inflammation, probably by promoting T regulatory immune response via an enhanced expression of TLR2, TLR4 and PD-L1 on DCs.

Eliminação de Dioctophyme renale pela urina em canino com dioctofimatose em rim esquerdo e cavidade abdominal - Primeiro relato no Rio Grande do Sul / Elimination of Dioctophyme renale to urine in canines with dioctophymosis in the left kidney and abdominal cavity - First report in Rio Grande do Sul

Dioctophyme renale é um parasito que afeta tanto animais como humanos e tem como órgão de eleição o rim direito. Relata-se o caso clínico-cirúrgico de um paciente canino com histórico de eliminação de três exemplares de D. renale pela urina e presença de parasitos no rim esquerdo e na cavidade abdominal. No Hospital de Clínicas Veterinárias da Universidade Federal de Pelotas, foram realizados exames pré-cirúrgicos, entre eles a ultrassonografia abdominal, que identificou a localização dos nematódeos, e o exame de Doppler pulsado dos vasos intrarrenais, que demonstrou aumento nos índices resistivos nas regiões avaliadas. O paciente foi conduzido para a realização de laparotomia exploratória, sendo removidos 23 parasitos da cavidade abdominal. A presença de D. renale causa graves lesões nos rins e nos demais órgãos da cavidade abdominal, sendo essencial o diagnóstico precoce e a remoção dos nematódeos para recuperação do paciente.(AU)

Dioctophyme renale is a parasite that attacks animals as well as humans and has the right kidney as a preferred target organ. This document reports the case of a clinical-chirurgical canine patient with a history of elimination of three D. renale samples through its urine and presence of parasites at its left kidney and abdominal cavity. Pre-cirurgical exams were performed at the Veterinary Hospital of the Federal University of Pelotas. Among these exams, abdominal ultrasonography identified the nematode's location and the Pulse-Doppler of the intrarenal vessels showed an increase in the resistive indices of the evaluated areas. An Exploratory Laparotomy was conducted with the patient who had 23 parasites removed from its abdominal cavity. The presence of D. renale causes severe lesions at kidneys and other organs of the abdominal cavity, for this reason early diagnosis and nematode´s removal are essential for the patient's recovery.(AU)

Strychnos pseudoquina A. St. Hil.: a Brazilian medicinal plant with promising in vitro antiherpes activity.

AIMS: To investigate the anti-HSV and anti-inflammatory effects of a standardized ethyl acetate extract (SEAE) prepared with the stem bark of Strychnos pseudoquina, along with two isolated compounds: quercetin 3-O-methyl ether (3MQ) and strychnobiflavone (SBF). METHODS AND RESULTS: The mechanisms of action were evaluated by different methodological strategies. SEAE and SBF affected the early stages of viral infection and reduced HSV-1 protein expression. Both flavonoids elicited a concentration-dependent inhibition of monocyte chemoattractant protein-1 (MCP-1), whereas 3MQ reduced the chemokine release more significantly than SBF. Conversely, both compounds stimulated the production of the cytokines TNF-α and IL-1-ß in LPS-stimulated cells, especially at the intermediate and the highest tested concentrations. CONCLUSIONS: SEAE and SBF interfered with various steps of HSV replication cycle, mainly adsorption, postadsorption and penetration, as well as with ß and γ viral proteins expression; moreover, a direct inactivation of viral particles was observed. Besides, both flavonoids inhibited MCP-1 selectively, a feature that may be beneficial for the development of new anti-HSV agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicated that the samples present anti-HSV and anti-inflammatory activities, at different levels, which is an interesting feature since cold and genital sores are accompanied by an inflammation process.

Severe Paracoccidioidomycosis in a 14-Year-Old Boy.

Paracoccidioidomycosis (PCM) is the most important systemic mycoses in Latin America. We describe a severe case of paracoccidioidomycosis in a 14-year-old boy, with a rapid disease progression. The fungal strain was isolated and inoculated into a T and/or B cell immunocompromised mice, which revealed a highly virulent strain. The case report presented herein emphasizes the importance of considering PCM in the differential diagnosis of patients with other infectious diseases in endemic areas and highlights a novel isolate.

In vitro and in vivo action of Piptadenia viridiflora (Kunth) Benth against Haemonchus contortus in sheep.

Anthelminthic resistant populations of Haemonchus contortus are a major problem in sheep rearing, but plant extracts may offer viable alternative treatments. In our preliminary studies, Piptadenia viridiflora was frequently selected by sheep grazing in the Cerrado. The present research evaluated its in vitro and in vivo anthelmintic activity. The HPLC chromatograms of P. viridiflora aqueous extract (AE) and ethanolic extract (EE) showed the presence of flavonoids. The total condensed tannin (proanthocyanidin) was 0.2 and 1.01% in AE and EE, respectively. In an egg hatching inhibition (EHI) test, the LC90 of AE was 2.4mg/mL, and, of EE, was 2.1mg/mL. After tannin extraction, higher EHI and lower LC90 were observed. In a larval development inhibition test, the LC90 of AE was 13.66mg/g of fecal culture. The highest dose of AE administered to mice (203.0mg/kg bw) was well tolerated, suggesting low toxicity. In vivo, AE was orally administered to lambs at 283mg/kg bw, and, at weeks one, two, and three post-treatment, the mean fecal egg count (FEC) was significantly lower than in untreated lambs (P<0.05). Blood parameters were normal and similar in untreated and treated sheep. For all lamb groups, the mean total serum protein was significantly higher at week two post-treatment than at other evaluated periods (P<0.05). Piptadenia viridiflora extracts had low condensed tannin content and exhibited high anthelminthic efficacy in vitro and significantly reduced FEC. Tannins were not shown to be the principal components affecting EHI, hence it is necessary to isolate and characterize the principal active P. viridiflora compounds, and to assess their possible synergism.

Clinical and immunological aspects and outcome of a Brazilian cohort of 414 patients with systemic lupus erythematosus (SLE): comparison between childhood-onset, adult-onset, and late-onset SLE.

The clinical expression of systemic lupus erythematosus (SLE) is influenced by genetic and environmental factors and therefore varies between ethnicities. Information on the epidemiology of SLE in Brazil is scarce and practically limited to studies conducted in socioeconomically developed regions (South and Southeast). The objective of this study was to describe the clinical and immunological aspects and outcome of a cohort of patients with SLE treated at a university hospital in northeastern Brazil and compare patterns related to age at onset: childhood (cSLE), adult (aSLE), and late (lSLE). A random sample of 414 records (women: 93.5%) were reviewed. The mean age at SLE onset and the mean disease duration were 28.9 ± 10.9 years and 10.2 ± 6.6 years, respectively. Most patients had aSLE (n = 338; 81.6%), followed by cSLE (n = 60; 14.5%) and lSLE (n = 16; 3.9%). The female/male ratio was 6.5:1 in cSLE and 16.8:1 in aSLE; in lSLE, all patients were female (p = 0.05). During follow-up, the cSLE group presented higher rates of nephritis (70% vs. 52.9% vs. 12.5%; p = 0.0001) and leuko/lymphopenia (61.7% vs. 43.8% vs. 56.2%; p = 0.02). No significant differences were found for anti-dsDNA, anti-Sm, and antiphospholipid antibodies. Treatment with immunosuppressants was significantly more common, and higher doses of prednisone were used, in cSLE. The prevalence of cardiovascular diseases were more frequent in lSLE (p = 0.03). No significant differences were found between the three groups with regard to mean damage accrual (SDI), remission, and mortality. Although cSLE presented higher rates of nephritis and leuko/lymphopenia, more frequent use of immunosuppressants and higher prednisone doses than aSLE and lSLE, the three groups did not differ significantly with regard to damage accrual, remission, and mortality.

An isolate of the nematophagous fungus Monacrosporium thaumasium for the control of cattle trichostrongyles in south-eastern Brazil.

A mycelial formulation in sodium alginate pellets of the nematophagous fungus Monacrosporium thaumasium (isolate NF34A) was assessed in the biological control of beef cattle trichostrongyles in tropical Brazil. Two groups of ten male Nellore calves aged 6 months, a fungus-treated group and a control group, were fed on a pasture of Brachiaria decumbens naturally infected with larvae of cattle trichostrongyles. The fungus-treated group received doses of sodium alginate mycelial pellets orally (1 g pellets (0.2 g fungus)/10 kg live weight) twice a week for 12 months. At the end of the study there was a significant reduction (P< 0.01) in the number of eggs per gram of faeces and coprocultures of the fungus-treated group--47.8% and 50.2%, respectively--in relation to the control group. There was a 47.3% reduction in herbage samples, collected up to 0-20 cm from faecal pats, between the fungus-treated and control groups, and a 58% reduction when the sampling distance was 20-40 cm from faecal pats (P< 0.01). The treatment with sodium alginate pellets containing the nematode-trapping fungus M. thaumasium reduced trichostrongyles in tropical south-eastern Brazil and could be an effective tool for the biological control of this parasitic nematode in beef cattle. However, in such a tropical climate with low rainfall the fungal viability can be reduced.

Fish distribution in watersheds of the eastern part of the Serra da Mantiqueira (state of São Paulo).

The present work aims to analyse jointly four contiguous watersheds in the eastern part of the Serra da Mantiqueira with respect to the distribution of fish fauna and physical structure. The watersheds of Piracuama river, Ribeirão Grande, Buenos and Guaratinguetá are located in Pindamonhangaba, Campos do Jordão and Guaratinguetá municipalities, in the state of São Paulo. Data were collected between the years 2001 and 2010, with collections distributed throughout the seasons, including sites located on the slope and piedmont. The limnological parameters analysed were different between watersheds and habitat structure was different only for the slope segments between Ribeirão Grande and Buenos watersheds, and between Ribeirão Grande and Piracuama watersheds. Thirty-five species of fish were caught, with the highest species richness found in the Ribeirão Grande watershed (30 species), followed by the Piracuama (23 species) and Buenos and Guaratinguetá (21 species each). The most abundant species in both the segments (slope and piedmont) was Trichomycterus itatiayae. The highest degree of species dominance occurred in the Guaratinguetá watershed and in the segment slope, being T. itatiayae the dominant species. Species diversity was lower at the slope than the piedmont, indicating variability in species abundance. Again T. itatiayae was the most abundant species in both segments, showing to be a well adapted species to these streams systems by presenting, as well as other species, morphological adaptations to the stream environment.

Parasitism by Paratanaisia bragai (Digenea, Eucotylidae) in commom waxbill (Estrilda astrild) / Parasitismo por Paratanaisia bragai em bico-de-lacre (Estrilda astrild)

O trematodeo digenético Paratanaisia bragai é descrito como parasito dos túbulos coletores renais e ureteres de aves. Esse helminto é usualmente pouco patogênico para a maioria dos hospedeiros; porém, em altas cargas parasitárias e em animais mais suscetíveis, pode causar sérios danos que resultam em apatia, perda de peso, desidratação e até mesmo a morte. O presente trabalho objetivou relatar pela primeira vez o trematoide P. bragai parasitando os rins de um passeriforme da espécie Estrilda astrild, conhecido popularmente como bico-de-lacre, de distribuição cosmopolita. A partir da análise histopatológica dos rins da ave, observou-se a presença dos vermes adultos e de ovos, grande dilatação dos ductos coletores renais, destruição e achatamento das células epiteliais de revestimento e, ao redor dos parasitos, observou-se moderada reação inflamatória. Os autores sugerem que a infecção pode ter sido acidental, uma vez que os moluscos terrestres que servem de hospedeiros intermediários para P. bragai não fazem parte da dieta natural descrita para os bico-de-lacre. Os resultados encontrados no presente estudo confirmaram um bico-de-lacre como possível hospedeiro de P. bragai, acrescentando essa espécie de ave e possivelmente todo o grupo dos passeriformes na epidemiologia da infecção por esse trematoide.(AU)

Characterization and cytotoxic activity of sulfated derivatives of polysaccharides from Agaricus brasiliensis.

Agaricus brasiliensis cell-wall polysaccharides isolated from fruiting body (FR) and mycelium (MI) and their respective sulfated derivatives (FR-S and MI-S) were chemically characterized using elemental analysis, TLC, FT-IR, NMR, HPLC, and thermal analysis. Cytotoxic activity was evaluated against A549 tumor cells by MTT and sulforhodamine assays. The average molecular weight (Mw) of FR and MI was estimated to be 609 and 310 kDa, respectively. FR-S (127 kDa) and MI-S (86 kDa) had lower Mw, probably due to hydrolysis occurring during the sulfation reaction. FR-S and MI-S presented ~14% sulfur content in elemental analysis. Sulfation of samples was characterized by the appearance of two new absorption bands at 1253 and 810 cm(-1) in the infrared spectra, related to S=O and C-S-O sulfate groups, respectively. Through (1)H and (13)C NMR analysis FR-S was characterized as a (1→6)-(1→3)-ß-D-glucan fully sulfated at C-4 and C-6 terminal and partially sulfated at C-6 of (1→3)-ß-D-glucan moiety. MI-S was shown to be a (1→3)-ß-D-gluco-(1→2)-ß-D-mannan, partially sulfated at C-2, C-3, C-4, and C-6, and fully sulfated at C-6 of the terminal residues. The combination of high degree of sulfation and low molecular weight was correlated with the increased cytotoxic activity (48 h of treatment) of both FR-S (EC50=605.6 µg/mL) and MI-S (EC50=342.1 µg/mL) compared to the non-sulfated polysaccharides FR and MI (EC50>1500 µg/mL).