Valuation of angiogenesis in bovine xenografts implanted in intracorporal sites of rabbits as models of in-vivo bioreactors / Avaliação da angiogênese em xenoenxertos bovinos implantados em sítios intracorpóreos de coelhos como modelos de biorreatores in vivo

The aim of this study was to evaluate neovascularization of bovine xenografts implanted in intracorporeal sites of rabbits (bioreactors). 30 rabbits were used, divided into 6 groups, according to the evaluation time (7, 15, 30, 45, and 60 days); each animal received xenogenic implants in 3 different intracorporeal sites (A1 - omentum bag; A2 - intermuscular space of quadriceps femoris; A3 - subperiosteal of ilium bone). Histological assessments graded the presence of angiogenesis, the number of inflammatory cells, newly formed bone tissue, and the presence of giant cells. Histological analyses showed intense angiogenesis in all implanted xenografts. Presence of inflammatory infiltrate and giant cells at the A1 implant site and presence of bone neoformation at the A3 implant site were noted. Degeneration of implants and formation of a fibrous capsule were noted. When comparing the interaction of the site with the days of evaluation, statistical analysis showed a significant difference (p≤0.05) in any time of neovascularization analysis. The vascular endothelial growth factor (VEGF) and inflammatory cells of the omentum in its structure, may have contributed to the greater presence of neovessels and inflammatory cells, a fact that may indicate functionality as a possible bone substitute.(AU)

O objetivo deste estudo foi avaliar a neovascularização de xenoenxertos bovinos implantados em sítios intracorpóreos de coelhos (biorreatores). Foram utilizados 30 coelhos, os quais foram divididos em seis grupos, de acordo com o tempo de avaliação (sete, 15, 30, 45 e 60 dias); cada animal recebeu implantes xenogênicos em três diferentes sítios intracorpóreos (A1 - bolsa de omento; A2 - espaço intermuscular do quadríceps femoral; A3 - subperiosteal do osso ílio). Avaliações histológicas classificaram a presença de angiogênese, o número de células inflamatórias, de tecido ósseo neoformado e a presença de células gigantes. As análises histológicas mostraram intensa angiogênese em todos os xenoenxertos implantados. Observou-se presença de infiltrado inflamatório e células gigantes no local do implante A1 e presença de neoformação óssea no local do implante A3. Ao mesmo tempo, a degeneração dos implantes e a formação de uma cápsula fibrosa foram observadas. Ao comparar a interação do local com os dias de avaliação, a análise estatística mostrou diferença significativa (P≤0,05) em qualquer momento da análise de neovascularização. O fator de crescimento endotelial vascular (VEGF) e as células inflamatórias do omento em sua estrutura podem ter contribuído para a maior presença de neovasos e células inflamatórias, fato que pode indicar funcionalidade como possível substituto ósseo.(AU)

Effects of dredging at Aratu port in All Saints Bay, Brazil: monitoring the metal content in water and sediments.

This work reports the chemical monitoring of saline water and sediments at Aratu port, All Saints Bay, Brazil, when subjected to a deep dragging process (DDP) over a 2-year period (2010-2012). The aim of this work was to detect which metals had possible deleterious effects on the biota. It analyzed percentage of silt + clay and TOC and metal concentrations (As, Cd, Cu, Cr, Ni, Pb, and Zn) before, during, and after the DDP. The determination of metal concentration in sediments used the following techniques: atomic absorption spectrometer with flame, also with hydrate generation, and cold vapor spectrophotometry. The DDP disturbed the concentrations of metals in sediments and water, but 15 months after the DDP, only Cu and Zn were detectable on the bottom saline water, and 20 months after the DDP, only Zn was detectable in water. This fact indicates that these metals were longer available for absorption by the marine biota with probable deleterious effect. Indeed, studies by others during the DDP in Aratu port, indicated the decrease from 50 to 40 of the number of microalgae species identified. On the other hand, in the sediments, 20 months after the DDP, the concentrations of As, Cr, Cu, Pb, and Hg, returned to the original values, but Ni and Zn did not return to their original values. Comparison with studies by others indicated that the sediments from dredging were inappropriate for disposal at sea. This work indicated that the metals Cu, Ni, and Zn are the most deleterious elements to the marine environment.

Cloning, expression, and antigenic characterization of recombinant protein of Mycoplasma gallisepticum expressed in Escherichia coli.

Mycoplasma gallisepticum (MG) is a member of the most important avian mycoplasmas, causing chronic respiratory disease in chickens and leading to important economic losses in the poultry industry. Recombinant technology represents a strategic approach used to achieve highly reliable and specific diagnostic tests in veterinary diseases control: in particular this aspect is crucial for confirming mycoplasma infection and for maintaining mycoplasma-free breeder flocks. In this study, we identified a component of the pyruvate dehydrogenase dihydrolipoamide acetyltransferase (i.e., E2) protein by 2-dimensional electrophoresis (2-DE), characterized it in immunoblotting assays, and analyzed its recombinant (r-E2) in a rec-ELISA test. For full-length protein expression in Escherichia coli (EC) a point mutation was introduced. A rabbit antiserum produced against r-E2 was tested in a Western Blot using different samples of Mycoplasma species. The results showed the applicability of site-directed mutagenesis, with a good yield of the r-E2 after purification. Also, anti-E2 serum reacted with all the tested MG strains showing no cross reaction with other mycoplasmas. The developed E2 ELISA test was capable of detecting MG antibodies in the sera examined. Those results demonstrate the antigenic stability of the E2 protein which could represent a recombinant antigen with potential diagnostic applications.

Evaluation of in vitro and in vivo adhesion and immunomodulatory effect of Lactobacillus species strains isolated from chickens.

This study aimed to characterize the in vitro and in vivo adhesion and immunomodulatory effect of Lactobacillus strains isolated from chickens. Lactobacillus samples isolated from 65-wk-old birds were identified by PCR; their adhesion was evaluated in vitro via basement membrane-type cell matrix and in vivo through carboxyfluorescein succinimidyl amino ester staining inoculation in 1-d-old birds and duodenum, jejunum, ileum, and cecum collections at 1, 4, 12, and 24 h after inoculation. The 5 best adhesive samples at the in vitro test formed a pool for total IgA and IgG measurement in sera and intestinal fluid. The birds were divided into groups by inoculation scheme: group 1 was treated with a pool of Lactobacillus spp. at 2-d-old and challenged 1 d later with Salmonella Enteritidis and then treated again with a pool of Lactobacillus spp. at 4 d of age; group 2 was treated with a pool of Lactobacillus spp. at 2 and 4 d of age; group 3 was challenged with Salmonella Enteritidis at 3 d of age; and group 4 was a negative control. Collections were taken at 7, 14, 21, 28, and 35 d after the first inoculation. The results suggest that basement membrane matrix use represents an important technique for triage of samples for subsequent in vivo evaluation and that carboxyfluorescein succinimidyl amino ester staining is efficient for identifying this bacterial characteristic. The Lactobacillus-treated groups (1 and 2) presented the highest IgA concentrations at the end of the experiment (12,054.6 and 10,568.4 ng/mL, respectively). The group 2 IgG values in intestinal fluid exceeded those of the other 3 groups (P < 0.05), peaking at 6.419 ng/mL. In most serum collections, the Lactobacillus-treated groups (1 and 2) did not differ significantly in IgG concentrations (P > 0.05), whereas group 3 presented the highest concentration of this antibody. It is concluded that there was greater adhesion of strains in the cecum and an important correlation between in vitro and in vivo results. These results also suggest the immunomodulatory action of Lactobacillus spp. in the chicken.

Transference in vitro of the resistance to the antimicrobials between Escherichia coli, Lactobacillus spp. and Salmonella enteritidis isolated from chickens / Transferência in vitro da resistência aos antimicrobianos entre Escherichia coli, Lactobacillus.spp e Salmonella enteritidis isoladas de frangos

The objective of this work was to verify the possibility of transference of resistance to the antimicrobials between bacteria that are in the present normal microbiota of chickens and Salmonella Enteritidis. Samples of Lactobacillus spp. (L. spp.), Salmonella Enteritidis (SE) and Escherichia coli (E. coli) previously isolated from chickens, selected after the test of sensitivity antimicrobial in vitro according the standard method (National Committee for Clinical Laboratory Standards) utilizing those with resistance and sensibility to the antimicrobials inductors, named donor and receptor bacteria, respectively were used. Antimicrobials inductors were utilized to stimulate the transference of resistance to the antimicrobials between the bacteria. The possibility of transference was verified from the E. coli resistant to the SE and L. spp. Transference of a sample of L. spp resistant to the antimicrobials inductors to the SE was also verified. It was only possible to verify the transference of the resistance to the antimicrobials inductor when the donor bacteria was the E. coli and the bacteria receptor was SE. In the present study we conclude that the transference of resistance to the antimicrobials between bacteria is possible, however, not all bacteria participate in that trial, not transmitting and neither acquiring this resistance.

O objetivo deste trabalho foi verificar a possibilidade de transferência de resistência aos antimicrobianos entre bactérias normais da microbiota de frangos e Salmonella Enteritidis. Utilizamos amostras de Lactobacillus spp. (L. spp.), Salmonella Enteritidis (SE) e Escherichia coli (E. coli) previamente isolados de frangos, selecionados após prova de sensibilidade antimicrobiana in vitro conforme metodologia padrão (Comitê Nacional para Padrões Clínicos de Laboratório). Utilizamos aqueles com resistência e sensibilidade aos antimicrobianos indutores, chamados de bactérias doadoras e receptoras, respectivamente. Os antimicrobianos indutores foram utilizados para estimular a transferência de resistência aos antimicrobianos entre as bactérias. A possibilidade de transferência foi verificada da E. coli resistente para a SE e L. spp. Também foi verificada a transferência de uma amostra de L. spp resistente aos antimicrobianos indutores para a SE. Só foi possível verificar a transferência da resistência aos antimicrobianos indutores quando a bactéria doadora foi a E. coli e a bactéria receptora foi a SE. No presente estudo concluímos que a transferência de resistência aos antimicrobianos entre bactérias é possível, mas nem todas as bactérias participam desse evento, não transmitindo e nem adquirindo esta resistência.

Modos de morrer na UTI pediátrica de um hospital terciário / Modes of death in PICU of a tertiary hospital

OBJETIVO: Determinar a prevalência dos diferentes modos de morrer e identificar limites terapêuticos em pacientes de uma Unidade de Tratamento Intensivo Pediátrica (UTIP) de hospital universitário. MÉTODOS: Estudo retrospectivo, baseado na revisäo de prontuários dos pacientes que morreram na UTIP do Hospital de Clínicas de Porto Alegre, no período de 1º julho de 1996 a 30 junho de 1997. Para modo de morrer, adotou-se critérios de: näo resposta às medidas de ressuscitaçäo cardiorrespiratória (NRRST), morte cerebral (MC), retirada/näo-adoçäo de medidas de suporte de vida (R/NASV) e decisäo de näo reanimar (DNR). Para causa de morte, adotou-se critério de falências de órgäos. RESULTADOS: Dos 61 óbitos ocorridos no período, entraram no estudo 44 pacientes, cuja mediana de idade foi 28 meses. Todos tiveram como causa de morte a falência de múltiplos órgäos. Vinte e seis pacientes (59 por cento) eram do grupo I (NRRST e MC), enquanto 18 (41 por cento) do grupo II (R/NASV e DNR). No grupo II, 83 por cento dos pacientes tinham doença crônica e/ou debilitante (p = 0,017; chi²). Os motivos de admissäo mais prevalentes foram necessidade de terapias de suporte (55 por cento), ventilatória e cardiocirculatória, sem diferença estatística entre os grupos. A mediana do tempo de permanência na UTI foi de 5 dias, e no hospital, de 11 dias, sem significância estatística entre os dois grupos. CONCLUSÖES: Observou-se alta prevalência de modos de morrer R/NASV e DNR nos pacientes de UTIP avaliados, sugerindo condutas de limitaçäo terapêutica para eles. Näo se conseguiu avaliar o nível de participaçäo da equipe e da família nesse processo de tomada de decisöes

[Modes of death in the PICU of a tertiary care hospital]. / Modos de morrer na UTI pediátrica de um hospital terciário.

BACKGROUND: To determine the prevalence of different modes of death and to identify therapeutic limitations in patients admitted to the Pediatric Intensive Care Unit (PICU) at a teaching hospital. METHODS: A retrospective study was carried out. Information was collected from the medical records of patients who died at the PICU of Hospital de Clínicas de Porto Alegre between July 1st, 1996 and June 30th, 1997. Four modes of death were considered: patient not responding to cardiopulmonary resuscitation methods (NRCPR), brain death (BD), withdrawal / withhold life-support measures (W/WLS) and decision not to resuscitate (DNR). For cause of death we employed the criterion of organ failure. RESULTS: Out of 61 deaths occurred during the period under analysis, 44 patients were included in the study. Patient age median was 28 months. The cause of death for all patients was multiple organ failure. Twenty-six patients (59%) were classified in group I (NRCPR and BD) and 18 (41%) in group II (W/WLS and DNR). Among patients of group II, 83% had a chronic and/or debilitating disease (p = 0.017; chi(2)). The prevalent reason for patient admission was the need for organ support (55%), both respiratory and cardiovascular. The median for duration of patient stay at the PICU was 5 days and at the hospital was 11 days. There was no statistically significant difference between the two groups in terms of reason for patient admission or duration of PICU/hospital stay. CONCLUSIONS: There was a high prevalence of W/WLS and DNR deaths among ICU patients, thus suggesting therapeutic limitation for them. We were unable to evaluate the level of participation by the medical team and by the family in this decision making process.