Trypsin IV or mesotrypsin and p23 cleave protease-activated receptors 1 and 2 to induce inflammation and hyperalgesia

Although principally produced by the pancreas to degrade dietary proteins in the intestine, trypsins are also expressed in the nervous system and in epithelial tissues, where they have diverse actions that could be mediated by protease-activated receptors (PARs). We examined the biological actions of human trypsin IV (or mesotrypsin) and rat p23, inhibitor-resistant forms of trypsin. The zymogens trypsinogen IV and pro-p23 were expressed in Escherichia coli and purified to apparent homogeneity. Enteropeptidase cleaved both zymogens, liberating active trypsin IV and p23, which were resistant to soybean trypsin inhibitor and aprotinin. Trypsin IV cleaved N-terminal fragments of PAR(1), PAR(2), and PAR(4) at sites that would expose the tethered ligand (PAR(1) = PAR(4) > PAR(2)). Trypsin IV increased [Ca(2+)](i) in transfected cells expressing human PAR(1) and PAR(2) with similar potencies (PAR(1), 0.5 microm; PAR(2), 0.6 microm). p23 also cleaved fragments of PAR(1) and PAR(2) and signaled to cells expressing these receptors. Trypsin IV and p23 increased [Ca(2+)](i) in rat dorsal root ganglion neurons that responded to capsaicin and which thus mediate neurogenic inflammation and nociception. Intraplantar injection of trypsin IV and p23 in mice induced edema and granulocyte infiltration, which were not observed in PAR (-/-)(1)(trypsin IV) and PAR (-/-)(2) (trypsin IV and p23) mice. Trypsin IV and p23 caused thermal hyperalgesia and mechanical allodynia and hyperalgesia in mice, and these effects were absent in PAR (-/-)(2) mice but maintained in PAR (-/-)(1) mice. Thus, trypsin IV and p23 are inhibitor-resistant trypsins that can cleave and activate PARs, causing PAR(1)- and PAR(2)-dependent inflammation and PAR(2)-dependent hyperalgesia.

CD8(+) T cell adjuvant effects of Salmonella FliCd flagellin in live vaccine vectors or as purified protein

Salmonella flagellin, the flagellum structural subunit, has received particular interest as a vaccine adjuvant conferring enhanced immunogenity to soluble proteins or peptides, both for activation of antibody and cellular immune responses. In the present study, we evaluated the Salmonella enterica FliCd flagellin as a T cell vaccine adjuvant using as model the 9-mer (SYVPSAEQI) synthetic H2(d)-restricted CD8(+) T cell-specific epitope (CS(280-288)) derived from the Plasmodium yoelii circumsporozoite (G) protein. The FliCd adjuvant effects were determined under two different conditions: (i) as recombinant flagella, expressed by orally delivered live S. Dublin vaccine strains expressing the target CS(280-288) peptide fused at the central hypervariable domain, and (ii) as purified protein in acellular vaccines in which flagellin was administered to mice either as a recombinant protein fused or admixed with the target CS(280-288) peptide. The results showed that CS(280-288)-specific cytotoxic CD8(+) T cells were primed when BALB/c mice were orally inoculated with the expressing the CS280-288 epitope S. Dublin vaccine strain. In contrast, mice immunized with purified FliCd admixed with the CS280-288 peptide and, to a lesser extent, fused with the target peptide developed specific cytotoxic CD8(+) T cell responses without the need of a heterologous booster immunization. The CD8(+) T cell adjuvant effects of flagellin, either fused or not with the target peptide, correlated with the in vivo activation of CD11c(+) dendritic cells. Taken together, the present results demonstrate that Salmonella flagellins are flexible adjuvant and induce adaptative immune responses when administered by different routes or vaccine formulations. (C) 2009 Elsevier Ltd. All rights reserved.

As propostas educacionais maranhenses no Império (1834-1889)

A instruçio p~blica no Maranhão sofreu certas mudanças, a partir do Ato Adicibnal (1834), que coiocou sob a responsabilidade das Administrações provinciais a direção do ensino p~blico, primario e secundario no Brasil. No s~culo passado, (XIX), no final dos anos 30 e durante os decênios de 40 e 50, houve, no Maranhão, um certo esforço do Estado, no sentido de exercer um contróle maior sobre a instrução provincial e de desenvolver ~ uma. maior clientela o ensino elementar. Nessa época, foram. pr~:))lr?v1dos pela Administração local os primeiros incentivos ã di fusão do ensino t~cnico e agrIcola; e reorganizado o ensino securidirio com a criação do Liceu Maranhense. Nas ~ltimas d~cadas do Imperio, entretanto, o Estado demonstrou um gra~ de desinteresse pelo desenvolvimento da educação escolar no Maranhão. As realizações oficiais dessa ~po~a, nos diver sos nIveis, ' foram insignificantes. A Administração Local, decretando o livre exeycIcio do ensino, em 1874, demonstrou legalmente o descaso a que relegou a instrução p~blica pr~ vincial. As tendências verificadas no ensino p~blico maranhense, nesse perIodo (1834-1889), for~m sobretudo, por particularidades do processo determinadas, polItico-econô mico marannense da ~poca. Nos primeiros decênios (final .de 30 : ê : · d~ca.das 40 ~. 50)', a sit~ação pó'Útiêa . ~aranhens.e · ;'.ar.act.eri:za·v~-. se por um grande empenho dos fazendeiros (fração da classe dominante no poder) em garantir a ordem e buscar a nia. No âmbito econômico, destacava-se um afã por hegem2. reformas materiais, visando ao soerguimento da economia mercantil. Ji nos Gltirnos dec~nios analisados, a situação p61Itico-ec2. n~mica traduzia-se por um maior fortalecimento do poder Político dos fazendeiros, mas com uma grave crise econômica. As condições sociais maranhenses nas primeiras décadas favo receram, pois, o surgimento de uma maior preocupação do Es tado pela educação escolar, enquanto que, na última fase, a situação político-econômica maranhense desestimulou um interesse maior do Estado pelo desenvolvimento do ensino público no Maranhão.

Efeito do uso de óleos vegetais, associados ou não a inseticida, no controle de bemisia tabaci (gennadius, 1889) e thrips tabaci (lindeman, 1888), em feijoeiro, na época das águas

The objective of this experiments was evaluate the control of Bemisia tabaci and Thrips tabaci, through the use of plant oils associated or not to insecticide in bean plants, besides verifying the consequences on yield. The experiment was carried out in the period of rainy seasons, by utilizing Carioca variety. The statistical design was the randomized blocks, by employing a 6x2 factorial scheme ( plant oil versus insecticide), totalizing 12 treatments and four replications. It was concluded thet application of plant oils without insecticide can be utilized on the control of the B. tabaci biotipo B, since their population and the symptoms of golden mosaic was similar to the treatment plant oils plus insecticide; among the plant oils Agrex (R) plant oil promoted a higher control of white fly. For T. tabaci the application of plant oil associated to insecticide provide with a smaller population index when compared to the treatments without this association. The yield was not affected by the population of B. tabaci biotipo B and T. tabaci when plant oils were utilized, associated or not to the insecticide.

A Família do tesouro: a monumentalização da família Albuquerque Maranhão e a luta pelo poder no Rio Grande do Norte (1889-1914)

The objective of this study is to discuss the process of building a family monumentalization Albuquerque Maranhão showed that both the traditional historiography of Rio Grande do Norte, represented by Tavares de Lyra, Rocha Pombo e Câmara Cascudo, as reflected in urban areas of Natal. To understand this process, we intend to analyze the production of the aforementioned authors as well as more recent studies, trying to discern or identify an attempt to link them to the family name to the history of Albuquerque Maranhão State, which ended up giving visibility to this group, making it the characters featured in the scenario of local history, investing them with a monumental character. In addition to historical analysis, we observe changes in the urban landscape of the city of Natal in the early twentieth century orchestrated by members of this family, which tied his line to public spaces for a new and modern city. Through this review, we will be able to realize that such practices turned out to be a stage of political disputes between Albuquerque Maranhão and opposition groups who were anxious to remove them both from the center of historical narratives on the Rio Grande do Norte, as well as the political space of the State environment exclusive domain of this group for nearly twenty years

Injuries caused by scorpionfishes and (Scorpaena plumieri Bloch, 1789 Scorpaena brasiliensis Cuvier 1829) in the Southwestern Atlantic Ocean (Brazilian coast): epidemiologic, clinic and therapeutic aspects of 23 stings in humans

Fishes of the family Scorpaenidae are responsible for severe injuries and occasionally deaths in humans around the world. The more venomous fishes on the Brazilian coast and in the Southwestern Atlantic region are classified in the genus Scorpaena (family Scorpaenidae). However, there are few studies on the venomous apparatus, the effects of the venom, or clinical aspects of human envenoming provoked by Atlantic scorpionfishes.In this communication, the authors present 23 accidents caused by scorpionfishes of the genus Scorpaena among fishermen, and report the species that provoked the injuries, the circumstances of contacts, the clinical aspects observed and the therapeutic measures utilized for control of the symptoms of the victims. The intense pain and the systemic findings observed in the patients were very frequent and we think that the injuries provoked by scorpionfishes should be considered the most important manifestations caused by venomous fishes of the East Atlantic Ocean. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Post-operative analgesic effects of butorphanol or firocoxib administered to dogs undergoing elective ovariohysterectomy

ObjectiveTo compare the post-operative analgesic effects of butorphanol or firocoxib in dogs undergoing ovariohysterectomy.Study designProspective, randomized, blinded, clinical trial.AnimalsTwenty-five dogs > 1 year of age.MethodsDogs received acepromazine intramuscularly (IM), 0.05 mg kg-1 and either butorphanol IM, 0.2 mg kg-1 (BG, n = 12) or firocoxib orally (PO), 5 mg kg-1 (FG, n = 13), approximately 30 minutes before induction of anesthesia with propofol. Anesthesia was maintained with isoflurane. Ovariohysterectomy was performed by the same surgeon. Pain scores using the dynamic and interactive visual analog scale (DIVAS) were performed before and at 1, 2, 3, 4, 6, 8 and 20 hours after the end of surgery by one observer, blinded to the treatment. Rescue analgesia was provided with morphine (0.5 mg kg-1) IM and firocoxib, 5 mg kg-1 (BG only) PO if DIVAS > 50. Groups were compared using paired t-tests and Fisher's exact test (p < 0.05). Data are presented as mean +/- SD.ResultsThe BG required significantly less propofol (BG: 2.6 +/- 0.59 mg kg-1; FG: 5.39 +/- 0.7 mg kg-1) (p < 0.05) but the anesthesia time was longer (BG: 14 +/- 6, FG: 10 +/- 4 minutes). There were no differences for body weight (BG: 7.9 +/- 5.0, FG: 11.5 +/- 4.6 kg), sedation scores, and surgery and extubation times (BG: 10 +/- 2, 8 +/- 5 minutes; FG: 9 +/- 3, 8 +/- 4 minutes, respectively) (p > 0.05). The FG had significantly lower pain scores than the BG at 1, 2 and 3 hours following surgery (p < 0.05). Rescue analgesia was administered to 11/12 (92%) and 2/13 (15%) dogs in the BG and FG, respectively (p < 0.05).Conclusion and clinical relevanceFirocoxib produced better post-operative analgesia than butorphanol. Firocoxib may be used as part of a multimodal analgesia protocol but may not be effective as a sole analgesic.

Postoperative analgesic effects of intravenous, intramuscular, subcutaneous or oral transmucosal buprenorphine administered to cats undergoing ovariohysterectomy

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

Ehrlichiosis in anemic, thrombocytopenic, or tick-infested dogs from a hospital population in South Brazil

Ehrlichia canis has a worldwide distribution, but clinical manifestations may vary geographically. We selected 129 dogs to determine prevalence of ehrlichiosis in dogs with anemia, thrombocytopenia, or ticks presented to a Veterinary Teaching Hospital in South Brazil. Of the 129 dogs, 68 carried the brown dog tick (Rhipicephalus sanguineus), 61 had thrombocytopenia (platelet count <150,000/μl), and 19 had anemia (PCV < 22%). Twenty dogs fulfilled more than one inclusion criteria. Ehrlichiosis was diagnosed by positive amplification of ehrlichial DNA by PCR using primers ECC and ECB that amplify a sequence of the 16S rRNA gene. Presence of E. canis was confirmed by cleavage of the amplified DNA using endonucleases HaeIII and AvaI. Fourteen of 68 (21%) dogs with ticks had ehrlichiosis, whereas 12 of 61 (20%) dogs presented with thrombocytopenia and 4 of 19 (21%) anemic dogs had ehrlichiosis. Similar results were obtained in dogs with thrombocytopenia and anemia (one of eight positive) and in dogs with thrombocytopenia and ticks (two of seven positive). All four dogs with anemia and ticks, and the dog that fulfilled all inclusion criteria yield no amplification of ehrlichial DNA by PCR. Based on our results, one in each five dogs infested by the brown dog tick, with anemia or thrombocytopenia had ehrlichosis. Contrary to widespread believe, ehrlichiosis was not the main cause for thrombocytopenia in our region. © 2003 Elsevier B.V. All rights reserved.

Usefulness of Mycobacterium tuberculosis molecular typing in a tuberculosis low-endemic agro-industrial setting of Brazil

To highlight the transmission and major phylogenetic clades of Mycobacterium tuberculosis, a retrospective study was carried out at two health facilities in a small agro-industrial area in São Paulo, Brazil, that has a low tuberculosis incidence rate. IS6110-RFLP and spoligotyping were performed on the isolates, with the former revealing that 31.3% (35/112) of strains were clustered. Epidemiological links were found in 16 of the 35 clustered patients and were associated with transmission among patients living in public housing. Spoligotyping grouped 62.8% of the strains. The T genetic family predominated among the isolates. Of interest is that five strains had a pattern characteristic of African or Asian origin (ST535), and two others were of the rare localized type ST1888 (BRA, VEN). In addition, three new types-1889, 1890, and 1891-were identified. Spoligotyping showed that some ST may be circulating to or from Brazil, and RFLP revealed ongoing transmission in inadequately ventilated public-housing buildings. This may point to a failure in tuberculosis control policy.

Influence of δβ-thalassemia or regulatory elements in individuals with increased fetal Hb levels in the São Paulo northwest population

Fetal hemoglobin (Hb F), formed by two alpha globin chains (α) and two gamma chains (γ) (α2 γ2), has reduced expression in adults, ranging from 0 to 1% of total hemoglobin. Increased levels of Hb F are due to mutations in the β-globin family, which cause hereditary persistence of fetal hemoglobin (HPFH) and delta-beta thalassemia (δβ-thalassemia).The control of the production takes place by the regulatory region and regions outside the β-globin family, among them 2q16, 6q23, 8q, and Xp22.2.The aims of this study were to determine the presence and frequency of two mutations for δβ-thalassemia, the XmnI polymorphism and β-globin haplotypes in healthy individuals with increased Hb F in the State of São Paulo. We analyzed 60 samples of peripheral blood of healthy adults, without complaints of anemia. The samples were separated into two groups according to Hb F level: group I - 34 samples with Hb F ranging from 2 to 15% and group II - 26 samples with Hb F over 15%. In relation to the polymorphisms examined, we found three heterozygous individuals (5%) for Spanish δβ-thalassemia, belonging to group I, whose Hb F levels were within the normal range.The Sicilian δβ-thalassemia mutation was not found, indicating the need to study other polymorphisms related to the increase of Hb F in adult life.The frequency of XmnI polymorphism was 33.3% and the mean Hb F levels were 15.48 ± 11.69%.The frequency observed in our study for this polymorphic site is higher than that found in the literature for healthy subjects.This polymorphism was more prevalent in individuals with Hb F levels below 15%. For four samples positive for this polymorphism, the Hb F levels were explained by the presence of HPFH and Spanish δβ-thalassemia mutations, so that the presence of the XmnI polymorphic site was not a determinant in the overexpression of γ-globin genes. Regarding β-globin haplotypes, 18 alleles and 27 distinct genotypic patterns were found.The pattern Atp1/Atp2 was the mostfrequent genotype (13.72%).Of the 18 alleles, 13 showed atypical patterns.The results show that the haplotype V was the most frequent (27.45%), followed by atypical Atp2 (13.72%) and Atp1 (11.76%), and that there was a higher correlation with the presence of HPFH and XmnI polymorphism.The high frequency of haplotype V in our samples and high frequency of atypical haplotypes may reflect a high rate of miscegenation in this population, suggesting an ethnic characteristic for the Brazilian population, requiring the evaluation of population genetic markers to corroborate this hypothesis. © FUNPEC-RP.

Ablation of the ability to control the right-to-left cardiac shunt does not affect oxygen uptake, specific dynamic action or growth in the rattlesnake Crotalus durissus

The morphologically undivided ventricle of the heart in non-crocodilian reptiles permits the mixing of oxygen-rich blood returning from the lungs and oxygen-poor blood from the systemic circulation. A possible functional significance for this intra-cardiac shunt has been debated for almost a century. Unilateral left vagotomy rendered the single effective pulmonary artery of the South American rattlesnake, Crotalus durissus, unable to adjust the magnitude of blood flow to the lung. The higher constant perfusion of the lung circulation and the incapability of adjusting the right-left shunt in left-denervated snakes persisted over time, providing a unique model for investigation of the long-term consequences of cardiac shunting in a squamate. Oxygen uptake recorded at rest and during spontaneous and forced activity was not affected by removing control of the cardiac shunt. Furthermore, metabolic rate and energetic balance during the post-prandial metabolic increment, plus the food conversion efficiency and growth rate, were all similarly unaffected. These results show that control of cardiac shunting is not associated with a clear functional advantage in adjusting metabolic rate, effectiveness of digestion or growth rates. © 2013. Published by The Company of Biologists Ltd.

As origens da teoria dos invariantes na Inglaterra e o Mécanique Analytique de Lagrange (1788)

Pós-graduação em Educação Matemática - IGCE