CD73 Downregulation Decreases In Vitro and In Vivo Glioblastoma Growth.

Glioblastoma is the worst and most common primary brain tumor. Here, we demonstrated the role of CD73, an enzyme responsible for adenosine (ADO) production, in glioblastoma progression. ADO increased glioma cell viability via A1 receptor sensitization. CD73 downregulation decreased glioma cell migration and invasion by reducing metalloproteinase-2 and vimentin expression and reduced cell proliferation by 40%, which was related to necrosis and sub-G1 phase blockage of cell cycle. Those effects also involved the stimulation of Akt/NF-kB pathways. Additionally, CD73 knockdown or enzyme inhibition potentiated temozolomide cytotoxic effect on glioma cells by decreasing the IC50 value and sensitizing cells to a non-cytotoxic drug concentration. CD73 inhibition also decreased in vivo rat glioblastoma progression. Delivery of siRNA-CD73 or APCP reduced tumor size by 45 and 40%, respectively, when compared with control. This effect was followed by a parallel 95% reduction of ADO levels in cerebrospinal fluid, indicating the role of extracellular ADO in in vivo glioma growth. Treatment did not induce systemic damage or mortality. Altogether, we conclude that CD73 is an interesting target for glioblastoma treatment and its inhibition may provide new opportunities to improve the treatment of brain tumors. Graphical Abstract ᅟ.

Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks.

The identification of pathogens of viral (Rotavirus, Coronavirus), parasitic (Toxocara spp.) and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi) origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5%) strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene) or intermediately virulent (vapB gene) profiles. Sixty-three (31.5%) strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%), A (25.4%) and B2 (22.2%). No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks

The identification of pathogens of viral (Rotavirus, Coronavirus), parasitic (Toxocara spp.) and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi) origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5%) strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene) or intermediately virulent (vapB gene) profiles. Sixty-three (31.5%) strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%), A (25.4%) and B2 (22.2%). No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

A novel triazolic naphthofuranquinone induces autophagy in reservosomes and impairment of mitosis in Trypanosoma cruzi.

Chagas' disease, caused by the protozoan Trypanosoma cruzi, represents a serious health problem in Latin America, and the available chemotherapy, which is based on 2 nitro-derivatives, is not satisfactory. In folk medicine, natural products including naphthoquinones have been employed for the treatment of different parasitic diseases. In the pursuit of alternative drugs for Chagas' disease, we investigated the mechanism of action of the triazolic naphthoquinone (TN; 2,2-dimethyl-3-(4-phenyl-1H-1,2,3-triazol-1-yl)-2,3-dihydronaphtho[1,2-b]furan-4,5-dione), which is the most active compound against T. cruzi trypomastigotes among a series of naphthofuranquinones. TN was active against the 3 parasite forms producing a dose-dependent inhibitory effect. In epimastigotes, TN induced reservosome disruption, flagellar blebbing, Golgi disorganization, the presence of cytosolic concentric membrane structures and abnormal multiflagellar parasites. The treatment also led to the appearance of well-developed endoplasmic reticulum profiles surrounding organelles that associated with an increase in monodansylcadaverine labelling, suggesting autophagy as part of the TN mechanism of action. Interestingly, no ultrastructural damage was detected in the mitochondria of naphthoquinone-treated epimastigotes. Flow cytometric analysis demonstrated an impairment of mitosis, an increase in ROS production and the maintenance of mitochondrial membrane potential. TN could be a good starting point in the investigation of a chemotherapeutic approach for the treatment of Chagas' disease.

Mastite bovina por Pasteurella multocida: estudo de nove casos / Bovine mastitis by Pasteurella multocida: a study of nine cases

The uncommon occurrence of bovine mastitis by Pasteurella multocida is reported in nine cows. Mammary pasteurellosis was predominantly diagnosed predominantly in clinical cases, in dairy cows milked in the presence of calves. Low interference of immunosuppressive or predispose factors and absence of pulmonary signs were observed in animals. In vitro multiple drug resistance of isolates was observed to three or more conventional antimicrobials, especially with use of tetracycline and gentamicin. Epidemiologic findings, clinical signs, and antimicrobial profile were discussed in nine cases of bovine mastitis caused by P. multocida.

Surto de mastite bovina causada por linhagens de Pseudomonas aeruginosa multirresistentes aos antimicrobianos / Outbreak of bovine mastitis caused by multiple drug resistant Pseudomonas aeruginosa strains

Uncommon outbreak of mastitis caused by Pseudomonas aeruginosa strains in 19 cows was reported.Enrofloxacin (73.7%) and danofloxacin were in vitro effective against those strains. In vitro multipledrug resistance of the isolated bacteria was observed to five or more conventional antimicrobials usedfor mammary therapy, including ampicillin (100.0%), cefalexin (100.0%), cloxacillin (100.0%),gentamicin (100.0%), penicillin/novobiocin (100.0%), cefoperazone (52.6%), tetracycline (100.0%),florfenicol (94.7%), and neomycin (73.7%). Epidemiological findings, clinical signs, in vitrosusceptibility profile, control measures, and public health risks were discussed in cows infected by P. aeruginosa.

Naphthoimidazoles promote different death phenotypes in Trypanosoma cruzi.

SUMMARY: In a screening of 65 derivatives of natural quinones using bloodstream trypomastigotes of Trypanosoma cruzi, the 3 naphthoimidazoles derived from beta-lapachone - N1, N2 and N3--were selected as the most active. Investigation of their mode of action led to the characterization of mitochondrion, reservosomes and DNA as their main targets, and stimulated further studies on death pathways. Ultrastructural analysis revealed both autophagic (autophagosomes) and apoptotic-like (membrane blebbing) phenotypes. Flow cytometry analysis showed, in N2-treated trypomastigotes, a small increase of phosphatidylserine exposure, and a large increase in the percentage of necrosis, caused by N1 or N2. These death phenotypes were not detected in treated epimastigotes. The strong increase in labelling of monodansyl cadaverine, the inhibition of the death process by wortmannin or 3-methyladenine, the overexpression of ATG genes in treated epimastigotes, together with ultrastructural evidence point to autophagy as the predominant phenotype induced by the naphthoimidazoles. However, there are other pathways occurring concomitantly with variable intensities, justifying the need to detail the molecular features involved.

Peracute bovine mastitis caused by Klebsiella pneumoniae / Mastite bovina hiperaguda causada por Klebsiella pneumoniae

Relata-se a ocorrência de graves sintomas de mastite hiperaguda em vaca, causada por Klebsiella pneumoniae, na terceira semana de lactação. Descrevem-se aspectos epidemiológicos, sintomas clínicos, procedimentos de diagnóstico microbiológico, conduta terapêutica e medidas de controle.

Serotonin in the nervous system of the head region of the land planarian Bipalium kewense.

The presence and distribution of serotonin (5-hydroxytryptamine, or 5-HT) in the head region of the land planarian Bipalium kewense has been investigated by an indirect immunofluorescence technique combined with confocal scanning laser microscopy (CSLM), and also by immunogold labeling at ultrastructural level. Serotonin immunoreactivity (IR) was restricted to elements of the nervous system, such as the cerebral ganglion, and the peripheral nerve net. Most of 5-HT-immunoreactive neurons are at the periphery of the brain; they were identified as unipolar, bipolar, and multipolar neurons. The ultrastructural results using immunogold labeling confirm the location of 5-HT within electron-dense vesicles (50-120 nm in diameter), clustered both in the cell bodies and in their processes. The intense 5-HT-IR herein demonstrated for B. kewense adds new data to the poorly studied nervous system of land planarians.

[Malaria treatment with artesunate (retocaps) in children of the Brazilian Amazon]. / Tratamento da malária com artesunate (retocaps) em crianças da Amazônia brasileira.

We evaluated the clinical and therapeutic response to artesunate retocaps in 32 children admitted to the Fundação de Medicina Tropical do Amazonas (Amazon Foundation of Tropical Medicine) with clinical characteristics of moderate and severe malaria. Of these, 29 were infected with P. falciparum and 3 with P. vivax. They improved clinically 24 hours after the beginning of therapy, with 33. 3% of patients without fever, and after 48 hours, 77.2% of the children had no fever. The monitoring of asexual forms of the parasites showed that on D2 (day 2 of treatment) 58.6% of children with P. falciparum infection had no more parasites in the blood stream, on D4 all children had negative slides both for P. falciparum and for P. vivax infection. In a long-term follow up, we found 66.6% recrudescence in P. falciparum patients. The results enabled the conclusion that artesunate retocaps are efficient in practice and their use rapidly reduces the parasitemia and improves the patients' clinical picture. However, in P. falciparum malaria the recrudescence rate was very high. We observed no side effects from this drug.

Fronto-nasal dysostosis, callosal agenesis, crossed-fused ectopia, tibial hemimelia, and preaxial polydactyly of feet: severe expression of the acrocallosal syndrome?

We report on a girl with frontonasal "dysostosis," callosal agenesis, crossed-fused ectopia, tibial hemimelia, and preaxial polydactyly of feet. This pattern of the developmental defects suggests a severe form of the acrocallosal syndrome. Implications for genetic counselling are discussed.

Acute respiratory viral infections in ambulatory children of urban northeast Brazil.

The morbidity of acute respiratory infections in young children and the role of respiratory viruses were evaluated in a 29-month household-based study in an impoverished urban population in Fortaleza, Brazil; subjects were 175 children less than 5 years of age in 63 families. Home visits were conducted three times weekly during which staff recorded the presence of respiratory and systemic symptoms and collected upper respiratory tract samples for viral isolation. A large and sustained burden of respiratory illness was observed, and respiratory viruses were isolated in 35% of the samples collected. Of the isolates, 45.6% were rhinoviruses, 16% parainfluenza viruses, 15.8% enteroviruses, 9.9% adenoviruses, 7.0% herpes simplex viruses, and 5.7% influenza viruses. The results indicate that poor children in northeast Brazil have a high prevalence of respiratory illness and that rhinovirus is the most frequent respiratory virus.

[Cardiac manifestations of hyperthyroidism in the elderly]. / Manifestações cardíacas do hipertireoidismo no idoso.

PURPOSE: To evaluate the cardiocirculatory abnormalities of hyperthyroidism in the elderly. PATIENTS AND METHODS: Twenty-four hyperthyroid patients, 18 women and six men, aged 60 to 87 (average 73.5) years were studied. Seventeen (70.9%) patients had associated cardiocirculatory diseases. The evaluation was made on clinical grounds complemented by electrocardiographic, radiologic, phonomechanocardiographic and echocardiographic examinations. RESULTS: Cardiocirculatory symptoms were observed in 17 (70.9%) patients and congestive heart failure in nine (37.5%) of them. The electrocardiogram was abnormal in 20 (83.3%) patients and the tachyarrhythmias were the commonest abnormality (62.5%). Eight (33.3%) patients had chronic atrial fibrillation and five (20.8%) had sinus tachycardia. There was no significant statistical difference on the electrocardiograms of patients with and without cardiocirculatory abnormalities. Cardiomegaly was significantly more prevalent in hyperthyroid patients, with (64.7%) or without (57.1%) cardiocirculatory abnormalities, than in normal elderly (23.9%). Left ventricular performance was studied in 14 patients through the systolic quotient and was found normal or high in 12 (85.7%). The percentage of fractional shortening (delta D%) was higher than 30 in all patients. None of the patients was found to have symmetric or asymmetric hypertrophic cardiomyopathy and mitral valve prolapse on echocardiogram. CONCLUSION: Hyperthyroidism in the elderly patient determines frequently cardiocirculatory abnormalities that may be misdiagnosed with those caused by the ageing process or by associated cardiopathies. This diagnosis should be suspected in all elderly patients having tachyarrhythmias and/or cardiac failure resistant to usual therapy, mainly in patients without clear cardiocirculatory pathology.

Teste tuberculinico e vacinacao BCG intradermica em servidores de uma empresa publica. / Tuberculin test and BCG intradermic vaccination in workerrs of a Public Company

Os autores descrevem o inicio da implantacao de programa de controle da tuberculose em Empresa Publica, no Municipio de Sao Paulo, mostrando os resultados do cadastramento tuberculinico seguido de aplicacao da vacina BCG intradermica nos "reatores negativos". Foram aplicados 8.016 testes tuberculinicos e 3.637 vacinas BCG intradermicas. O indice de infeccao tuberculosa situou-se ao redor de 39%. E tambem discutida a introducao da vacina BCG intradermica no exame pre-admissional