An Oral Vaccine Based on U-Omp19 Induces Protection against B. abortus Mucosal Challenge by Inducing an Adaptive IL-17 Immune Response in Mice

As Brucella infections occur mainly through mucosal surfaces, the development of mucosal administered vaccines could be radical for the control of brucellosis. In this work we evaluated the potential of Brucella abortus 19 kDa outer membrane protein (U-Omp19) as an edible subunit vaccine against brucellosis. We investigated the protective immune response elicited against oral B. abortus infection after vaccination of mice with leaves from transgenic plants expressing U-Omp19; or with plant-made or E. coli-made purified U-Omp19. All tested U-Omp19 formulations induced protection against Brucella when orally administered without the need of adjuvants. U-Omp19 also induced protection against a systemic challenge when parenterally administered. This built-in adjuvant ability of U-Omp19 was independent of TLR4 and could be explained at least in part by its capability to activate dendritic cells in vivo. While unadjuvanted U-Omp19 intraperitoneally administered induced a specific Th1 response, following U-Omp19 oral delivery a mixed specific Th1-Th17 response was induced. Depletion of CD4+ T cells in mice orally vaccinated with U-Omp19 resulted in a loss of the elicited protection, indicating that this cell type mediates immune protection. The role of IL-17 against Brucella infection has never been explored. In this study, we determined that if IL-17A was neutralized in vivo during the challenge period, the mucosal U-Omp19 vaccine did not confer mucosal protection. On the contrary, IL-17A neutralization during the infection did not influence at all the subsistence and growth of this bacterium in PBS-immunized mice. All together, our results indicate that an oral unadjuvanted vaccine based on U-Omp19 induces protection against a mucosal challenge with Brucella abortus by inducing an adaptive IL-17 immune response. They also indicate different and important new aspects i) IL-17 does not contribute to reduce the bacterial burden in non vaccinated mice and ii) IL-17 plays a central role in vaccine mediated anti-Brucella mucosal immunity

Hemodynamic effects and electrocardiographics of the acute intoxication with bupivacaine, levobupivacaine and 50% enantiometric excess mixture : an experimental study in pigs

Orientador: Artur UdelsmannDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: Até recentemente, a bupivacaína tem sido o anestésico de escolha nos bloqueios locorregionais em razão da qualidade da anestesia proporcionada e pela sua duração. Apesar disso, sua toxicidade cardiovascular preocupa os anestesiologistas que procuram novas opções farmacológicas com menor grau desse inconveniente. Uma destas é o seu isômero levógiro, a levobupivacaína, que por uma menor afinidade aos receptores dos canais de sódio da célula cardíaca, seria menos cardiotóxica. Em nosso meio está disponível a apresentação contendo 75% do isômero levógiro e 25% do isômero dextrógiro, denominada mistura com excesso enantiomérico de 50%. O objetivo desse estudo foi avaliar as repercussões hemodinâmicas da injeção intravascular de dose tóxica desses três agentes, buscando encontrar qual deles tem menor impacto em caso de acidente. Suínos da raça Large White foram anestesiados com tiopental, intubados e ventilados mecanicamente, sendo em seguida instalada monitorização hemodinâmica com cateter de Swan-Ganz e pressão invasiva para estudo das variáveis hemodinâmicas. Após repouso, foram divididos aleatoriamente em três grupos e realizada intoxicação duplamente encoberta com um dos agentes na dose de 4 mg.kg-1. Os resultados hemodinâmicos foram avaliados durante 30 minutos. Aos resultados foram aplicados testes estatísticos para comparação entre os grupos. A mistura com excesso enantiomérico de 50% e a levobupivacaína causaram maiores repercussões hemodinâmicas do que a mistura racêmica, sendo estas mais pronunciadas com o primeiro agente. Estes resultados se opõem aos encontrados em humanos, particularmente quando da utilização do isômero levógiro puro, mas estão de acordo com resultados recentes também em animais. Extrapolar dados obtidos em suínos para seres humanos exige muita cautela e novos estudos são necessários. Em suínos, a mistura com excesso enantiomérico de 50% particularmente, e a levobupivacaína mostraram-se mais tóxicas quando administradas por via venosa do que a bupivacaína racêmicaAbstract: Until recently, bupivacaine had been the anesthetic of choice for loco-regional blocks due to the quality and duration of the anesthesia. But its cardiovascular toxicity is a source of concern for anesthesiologists who seek new pharmacological options with a smaller degree ofthis problem. Its levorotatory isomer, levobupivacaine, that would be less cardiotoxicitydue a smaller affinity for the receptorsof the sodium channels of the cardiac cell, is one of these options. In Brazil, a presentation containing 75% of the levorotatory isomer and 25% of the destrorotatory isomer, called 50 % enantiomeric excess mixture is available. The aim of this studywas to evaluate thr hemodynamic repercurssions of the intravascular injectionof a toxic dose of those three agents to determine wich one hás the least impact in the case of na accident. Large White pigs were anesthetized with tiopental, intubated, and placed on mechanical ventilation. Hemodynamic monitoring was achieved with Swan-Ganz catheter and invasive blood pressure. After period of rest, the animals were randomlydivided in three groups. The intoxication was performed, on a double-blind fashion, with 4 mg. Kg -1 of one of the drugs. Hemodynamic parameters were evaluate during 30 minutes. Analytical tests were used to compare the results among the groups. The 50 % enantiomeric excess mixture and levobupivacaine hás greater hemadynamic repercussions than the racemic mixture, which were more pronounced with the first drug. These results go against those found in humans, especially regarding the pure levarotatoty isomer, but are similar to recent results reported in animals. One should be careful when extrapolating the data obtained in pigs to humans and further studies are necessary. In pigs, the 50% enantiomeric excess mixture, in particular, and levobupivacaine were more toxic when administered intravenously than racemic bupivacaineMestradoCirurgiaMestre em Cirurgi

Electroencephalographic responses to a noxious surgical stimulus in mules, horses, and ponies

The aim of this study was to investigate the electroencephalographic (EEG) response of equidae to a castration stimulus. Study 1 included 11 mules (2 1/2-8 years; 230-315 kg) and 11 horses (1 1/2-3 1/2 years; 315-480 kg); study 2 included four ponies (15-17 months; 176-229 kg). They were castrated under halothane anesthesia after acepromazine premedication (IV [study 1] and intramuscular [study 2]) and thiopental anesthetic induction. Animals were castrated using a semiclosed technique (study 1) and a closed technique (study 2). Raw EEG data were analyzed and the EEG variables, median frequency (F50), total power (Ptot), and spectral edge frequency (F95), were derived using standard techniques at skin incision (skin) and emasculation (emasc) time points. Baseline values of F50, Ptot, and F95 for each animal were used to calculate percentage change from baseline at skin incision and emasculation. Differences were observed in Ptot and F50 data between hemispheres in horses but not mules (study 1) and in one pony (study 2). A response to castration (&gt;10% change relative to baseline) was observed in eight horses (73% of animals) and four mules (36% of animals) for F50 and nine horses (82%) and four mules (36%) for Ptot. No changes in F95 data were observed in any animal in study 1. Responses to castration were observed in three ponies (75% of animals) for F50, one pony (25%) for F95, and all ponies for Ptot. Alteration of acepromazine administration and castration technique produced a protocol that identified changes in EEG frequency and power in response to castration.</p

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). 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

ObjectiveTo compare the postoperative analgesic effects of intravenous (IV), intramuscular (IM), subcutaneous (SC) or oral transmucosal (OTM) buprenorphine administered to cats undergoing ovariohysterectomy.Study designRandomized, prospective and blinded clinical trial.Animals100 female cats.MethodsCats were assigned to receive 0.01 mg kg-1 of buprenorphine administered by the IV, IM, SC or OTM route (n = 25/group). Buprenorphine was made up to 0.3 mL with 0.9% saline. DIVAS (0-100 mm) and simple descriptive scale (SDS) (from 0 to 4) pain and sedation scores were assigned to each cat before and 1, 2, 3, 4, 6, 8, 12 and 24 hours after ovariohysterectomy. Buprenorphine and carprofen were administered for rescue analgesia. Data were analyzed using anova and Fisher's exact test (p 0.05). There were no significant differences between groups for sedation scores at any time. SDS pain scores did not detect any differences between groups (p > 0.05). DIVAS pain scores after OTM administration were significantly higher than IV and IM administration at 1 hour and at 3, 4, 6, 8 and 12 hours, respectively (p < 0.05). DIVAS pain scores after SC administration were significantly higher than IV and IM administration at 2 hours and at 2, 3, 4, 8, 12 and 24 hours (p < 0.05), respectively. Six, four, 13 and 17 cats that received IV, IM, SC and OTM buprenorphine required rescue analgesia, respectively. There was a significantly higher incidence of treatment failure in cats that received SC and OTM buprenorphine compared with cats that received IV and IM buprenorphine (p < 0.05).Conclusions and Clinical relevanceIV and IM administration of buprenorphine provided better postoperative analgesia than SC or OTM administration of the drug and these routes of administration should be preferred when buprenorphine is administered to cats.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Biomechanical and histologic analysis in aortic endoprosthesis using fibrin glue

Background: The absence of incorporation between endoprosthesis (EP) and the arterial wall may lead to device migration and endoleaks around the stent graft. Alternatives have been tested aiming to improve this incorporation. Fibrin glue is used in many operating procedures promoting adhesion and tissue regeneration; however, its use to improve EP incorporation by arteries is unknown.Objective: The objective of this study was to analyze dislodgement forces needed to extract the EPs implanted in pig aorta, compare different oversizing and fibrin glue injections, and to analyze histologic changes among groups.Methods: Straight EPs were implanted in the thoracic aorta of pigs using 10% oversizing plus fibrin glue in the interface between the EP and the artery (group 1), using 20% oversizing (group 2), and 10% oversizing (group 3). Fourteen days after the implant, the animals were killed to enable biomechanical analysis of the EP and to verify histologic changes of the aortic wall and its interface with the EP.Results: Group 1 showed a dislodgement force of 21.9 +/- 5.3 Newton (N) being higher than the other groups and statistically significant when compared to group 3 (15.6 +/- 3.6N), P = .003%. Group 2 had a higher dislodgement force and statistically more significant than group 3 (19.5 +/- 7.8N). Histologic analysis showed tissue reaction with inflammatory cells and fibroblasts higher in group 1 and group 2 compared to group 3.Conclusion:This study reports a large animal survival model of thoracic aortic stent graft placement by testing the impact of fibrin glue on EP incorporation. Compared to oversizing alone, fibrin glue placed between the stent graft and the arterial wall increases EP incorporation. Additional studies are needed to determine the potential utility of fibrin glue in the setting of human arterial endografts. (J Vase Surg 2011;53:1368-74.

Genetic risk factors for nonsyndromic cleft lip with or without cleft palate in a brazilian population with high african ancestry

Nonsyndromic cleft lip with or without cleft palate (NSCL +/- P) is the most common orofacial birth defect, exhibiting variable prevalence around the world, often attributed to ethnic and environmental differences. Linkage analyses and genome-wide association studies have identified several genomic susceptibility regions for NSCL +/- P, mostly in European-derived or Asian populations. Genetic predisposition to NSCL +/- P is ethnicity-dependent, and the genetic basis of susceptibility to NSCL +/- P likely varies among populations. The population of Brazil is highly admixed, with highly variable ancestry; thus, the genetic determinants of NSCL +/- P susceptibility may be quite different. This study tested association of 8 single-nucleotide polymorphisms (SNPs), previously identified by genome-wide studies in other populations, with NSCL +/- P in a Brazilian population with high African ancestry. SNPs rs560426, rs642961, rs1530300, rs987525, rs3758249, rs7078160, rs17085106, and rs13041247 were genotyped in 293 Brazilian patients with NSCL +/- P and 352 unaffected Brazilian controls. Each sample was also genotyped for 40 biallelic short insertion/deletion polymorphic markers to characterize genetic ancestry. The average African ancestry background was 31.1% for the NSCL +/- P group and 36.7% for the control group. After adjustment for ancestry and multiple testing, the minor alleles of rs3758249 (OR: 1.58, 95% CI: 1.25-2.01, P=0.0001) and rs7078160 (OR: 1.59, 95% CI: 1.21-2.07, P=0.0002) were significantly associated with risk of NSCL +/- P. Polymorphisms located in IRF6 (rs642961) and 8q24 (rs1530300 and rs987525) showed marginal associations in this Brazilian population with high African ancestry. These results indicate that rs3758249 at 9q22 and rs7078160 at 10q25.3 represent risk loci for NSCL +/- P in the Brazilian population with high African ancestry1671023442349CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DA BAHIA - FAPESBFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP470606/2011-8; 484292/2013-700007/014470606/2011-8; 484292/2013-

Neotropical freshwater fisheries : A dataset of occurrence and abundance of freshwater fishes in the Neotropics

The Neotropical region hosts 4225 freshwater fish species, ranking first among the world's most diverse regions for freshwater fishes. Our NEOTROPICAL FRESHWATER FISHES data set is the first to produce a large-scale Neotropical freshwater fish inventory, covering the entire Neotropical region from Mexico and the Caribbean in the north to the southern limits in Argentina, Paraguay, Chile, and Uruguay. We compiled 185,787 distribution records, with unique georeferenced coordinates, for the 4225 species, represented by occurrence and abundance data. The number of species for the most numerous orders are as follows: Characiformes (1289), Siluriformes (1384), Cichliformes (354), Cyprinodontiformes (245), and Gymnotiformes (135). The most recorded species was the characid Astyanax fasciatus (4696 records). We registered 116,802 distribution records for native species, compared to 1802 distribution records for nonnative species. The main aim of the NEOTROPICAL FRESHWATER FISHES data set was to make these occurrence and abundance data accessible for international researchers to develop ecological and macroecological studies, from local to regional scales, with focal fish species, families, or orders. We anticipate that the NEOTROPICAL FRESHWATER FISHES data set will be valuable for studies on a wide range of ecological processes, such as trophic cascades, fishery pressure, the effects of habitat loss and fragmentation, and the impacts of species invasion and climate change. There are no copyright restrictions on the data, and please cite this data paper when using the data in publications

Epidemiology and outcomes of hospital-acquired bloodstream infections in intensive care unit patients: the EUROBACT-2 international cohort study

Purpose In the critically ill, hospital-acquired bloodstream infections (HA-BSI) are associated with significant mortality. Granular data are required for optimizing management, and developing guidelines and clinical trials. Methods We carried out a prospective international cohort study of adult patients (≥ 18 years of age) with HA-BSI treated in intensive care units (ICUs) between June 2019 and February 2021. Results 2600 patients from 333 ICUs in 52 countries were included. 78% HA-BSI were ICU-acquired. Median Sequential Organ Failure Assessment (SOFA) score was 8 [IQR 5; 11] at HA-BSI diagnosis. Most frequent sources of infection included pneumonia (26.7%) and intravascular catheters (26.4%). Most frequent pathogens were Gram-negative bacteria (59.0%), predominantly Klebsiella spp. (27.9%), Acinetobacter spp. (20.3%), Escherichia coli (15.8%), and Pseudomonas spp. (14.3%). Carbapenem resistance was present in 37.8%, 84.6%, 7.4%, and 33.2%, respectively. Difficult-to-treat resistance (DTR) was present in 23.5% and pan-drug resistance in 1.5%. Antimicrobial therapy was deemed adequate within 24 h for 51.5%. Antimicrobial resistance was associated with longer delays to adequate antimicrobial therapy. Source control was needed in 52.5% but not achieved in 18.2%. Mortality was 37.1%, and only 16.1% had been discharged alive from hospital by day-28. Conclusions HA-BSI was frequently caused by Gram-negative, carbapenem-resistant and DTR pathogens. Antimicrobial resistance led to delays in adequate antimicrobial therapy. Mortality was high, and at day-28 only a minority of the patients were discharged alive from the hospital. Prevention of antimicrobial resistance and focusing on adequate antimicrobial therapy and source control are important to optimize patient management and outcomes