Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and Phosphatases.

Intestinal ischemia reperfusion injury (iIRI) is a severe clinical condition presenting high morbidity and mortality worldwide. Some of the systemic consequences of IRI can be prevented by applying ischemic preconditioning (IPC), a series of short ischemia/reperfusion events preceding the major ischemia. Although neutrophils are key players in the pathophysiology of ischemic injuries, neither the dysregulation presented by these cells in iIRI nor the protective effect of iIPC have their regulation mechanisms fully understood. Protein phosphorylation, as well as the regulation of the respective phosphatases and kinases are responsible for regulating a large number of cellular functions in the inflammatory response. Moreover, in previous work we found hydrolases and transferases to be modulated in iIR and iIPC, suggesting the possible involvement of phosphatases and kinases in the process. Therefore, in the present study, we analyzed the phosphoproteome of neutrophils from rats submitted to mesenteric ischemia and reperfusion, either submitted or not to IPC, compared to quiescent controls and sham laparotomy. Proteomic analysis was performed by multi-step enrichment of phosphopeptides, isobaric labeling, and LC-MS/MS analysis. Bioinformatics was used to determine phosphosite and phosphopeptide abundance and clustering, as well as kinases and phosphatases sites and domains. We found that most of the phosphorylation-regulated proteins are involved in apoptosis and migration, and most of the regulatory kinases belong to CAMK and CMGC families. An interesting finding revealed groups of proteins that are modulated by iIR, but such modulation can be prevented by iIPC. Among the regulated proteins related to the iIPC protective effect, Vamp8 and Inpp5d/Ship are discussed as possible candidates for control of the iIR damage.

Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome.

Intestinal ischemia and reperfusion injury is a model system of possible consequences of severe trauma and surgery, which might result into tissue dysfunction and organ failure. Neutrophils contribute to the injuries preceded by ischemia and reperfusion. However, the mechanisms by which intestinal ischemia and reperfusion stimulate and activate circulating neutrophils is still not clear. In this work, we used proteomics approach to explore the underlying regulated mechanisms in Wistar rat neutrophils after ischemia and reperfusion. We isolated neutrophils from three different biological groups; control, sham laparotomy, and intestinal ischemia/reperfusion. In the workflow, we included iTRAQ-labeling quantification and peptide fractionation using HILIC prior to LC-MS/MS analysis. From proteomic analysis, we identified 2,045 proteins in total that were grouped into five different clusters based on their regulation trend between the experimental groups. A total of 417 proteins were found as significantly regulated in at least one of the analyzed conditions. Interestingly, the enzyme prediction analysis revealed that ischemia/reperfusion significantly reduced the relative abundance of most of the antioxidant and pro-survival molecules to cause more tissue damage and ROS production whereas some of the significantly up regulated enzymes were involved in cytoskeletal rearrangement, adhesion and migration. Clusters based KEGG pathways analysis revealed high motility, phagocytosis, directional migration, and activation of the cytoskeletal machinery in neutrophils after ischemia and reperfusion. Increased ROS production and decreased phagocytosis were experimentally validated by microscopy assays. Taken together, our findings provide a characterization of the rat neutrophil response to intestinal ischemia and reperfusion and the possible mechanisms involved in the tissue injury by neutrophils after intestinal ischemia and reperfusion.

High performance mass spectrometry based proteomics reveals enzyme and signaling pathway regulation in neutrophils during the early stage of surgical trauma.

PURPOSE: In clinical conditions trauma is associated with high mortality and morbidity. Neutrophils play a key role in the development of multiple organ failure after trauma EXPERIMENTAL DESIGN: To have a detailed understanding of the neutrophil activation at primary stages after trauma, neutrophils are isolated from control and surgical trauma rats in this study. Extracted proteins are analyzed using nano liquid chromatography coupled with tandem mass spectrometry. RESULTS: A total of 2924 rat neutrophil proteins are identified in our analysis, of which 393 are found differentially regulated between control and trauma groups. By using functional pathways analysis of the 190 proteins up-regulated in surgical trauma, we found proteins related to transcription initiation and protein biosynthesis. On the other hand, among the 203 proteins down-regulated in surgical trauma we found enrichment for proteins of the immune response, proteasome degradation and actin cytoskeleton. Overall, enzyme prediction analysis revealed that regulated enzymes are directly involved in neutrophil apoptosis, directional migration and chemotaxis. Our observations are then confirmed by in silico protein-protein interaction analysis. CONCLUSIONS AND CLINICAL RELEVANCE: Collectively, our results reveal that neutrophils drastically regulate their biochemical pathways after the early stages of surgical trauma, showing lower activity. This implies higher susceptibility of the trauma patients to infection and bystander tissues damage.

Ischemic preconditioning modifies mortality and inflammatory response.

PURPOSE: To evaluate the effect of ischemic preconditioning on mortality, inflammatory mediators and oxidative stress after intestinal ischemia and reperfusion. METHODS: Male Wistar rats were allocated according to the period of ischemia with or without ischemic preconditioning which consist on clamping the superior mesenteric artery for 10 minutes followed by reperfusion for 10 minutes before the sustained ischemia period. Mortality was assessed in Phase 1 study, and the CINC-1, CINC-2 and MDA levels in the lungs were analyzed in Phase 2. RESULTS: Mortality was lower in the ischemic preconditioning group subjected to 90 minutes of ischemia compared to the group without ischemic preconditioning (I-90: 50% and IPC-90: 15%, p=0.018), and it was lower in the ischemic preconditioning group as a whole compared to the groups without ischemic preconditioning (IPC-14% and I=30%, p=0.006). Lower levels of MDA, CINC-1, and CINC-2 were observed in the animals that were subjected to ischemic preconditioning compared to the animals that were not (MDA: I-45=1.23 nmol/mg protein, and IPC-45=0.62 nmol/mg protein, p=0.0333; CINC-1: I-45=0.82 ng/mL and IPC-45=0.67 ng/mL, p=0.041; CINC-2: I-45=0.52 ng/mL and IPC-45=0.35 ng/mL, p=0.032). CONCLUSION: Ischemic preconditioning reduces mortality, inflammatory process and oxidative stress in rats subjected to intestinal ischemia and reperfusion.

Ischemic preconditioning modifies mortality and inflammatory response

PURPOSE: To evaluate the effect of ischemic preconditioning on mortality, inflammatory mediators and oxidative stress after intestinal ischemia and reperfusion. METHODS: Male Wistar rats were allocated according to the period of ischemia with or without ischemic preconditioning which consist on clamping the superior mesenteric artery for 10 minutes followed by reperfusion for 10 minutes before the sustained ischemia period. Mortality was assessed in Phase 1 study, and the CINC-1, CINC-2 and MDA levels in the lungs were analyzed in Phase 2. RESULTS: Mortality was lower in the ischemic preconditioning group subjected to 90 minutes of ischemia compared to the group without ischemic preconditioning (I-90: 50% and IPC-90: 15%, p=0.018), and it was lower in the ischemic preconditioning group as a whole compared to the groups without ischemic preconditioning (IPC-14% and I=30%, p=0.006). Lower levels of MDA, CINC-1, and CINC-2 were observed in the animals that were subjected to ischemic preconditioning compared to the animals that were not (MDA: I-45=1.23 nmol/mg protein, and IPC-45=0.62 nmol/mg protein, p=0.0333; CINC-1: I-45=0.82 ng/mL and IPC-45=0.67 ng/mL, p=0.041; CINC-2: I-45=0.52 ng/mL and IPC-45=0.35 ng/mL, p=0.032). CONCLUSION: Ischemic preconditioning reduces mortality, inflammatory process and oxidative stress in rats subjected to intestinal ischemia and reperfusion.

Evaluation of the effects of ischemic preconditioning on the hematological parameters of rats subjected to intestinal ischemia and reperfusion.

OBJECTIVES: Intestinal ischemia/reperfusion often leads to acute lung injury and multiple organ failure. Ischemic preconditioning is protective in nature and reduces tissue injuries in animal and human models. Although hematimetric parameters are widely used as diagnostic tools, there is no report of the influence of intestinal ischemia/reperfusion and ischemic preconditioning on such parameters. We evaluated the hematological changes during ischemia/reperfusion and preconditioning in rats. METHODS: Forty healthy rats were divided into four groups: control, laparotomy, intestinal ischemia/reperfusion and ischemic preconditioning. The intestinal ischemia/reperfusion group received 45 min of superior mesenteric artery occlusion, while the ischemic preconditioning group received 10 min of short ischemia and reperfusion before 45 min of prolonged occlusion. A cell counter was used to analyze blood obtained from rats before and after the surgical procedures and the hematological results were compared among the groups. RESULTS: The results showed significant differences in hematimetric parameters among the groups. The parameters that showed significant differences included lymphocyte, white blood cells and granulocyte counts; hematocrit; mean corpuscular hemoglobin concentration; red cell deviation width; platelet count; mean platelet volume; plateletcrit and platelet distribution width. CONCLUSION: The most remarkable parameters were those related to leukocytes and platelets. Some of the data, including the lymphocyte and granulocytes counts, suggest that ischemic preconditioning attenuates the effect of intestinal ischemia/reperfusion on circulating blood cells. Our work contributes to a better understanding of the hematological responses after intestinal ischemia/reperfusion and IPC, and the present findings may also be used as predictive values.

Evaluation of the effects of ischemic preconditioning on the hematological parameters of rats subjected to intestinal ischemia and reperfusion

OBJECTIVES: Intestinal ischemia/reperfusion often leads to acute lung injury and multiple organ failure. Ischemic preconditioning is protective in nature and reduces tissue injuries in animal and human models. Although hematimetric parameters are widely used as diagnostic tools, there is no report of the influence of intestinal ischemia/reperfusion and ischemic preconditioning on such parameters. We evaluated the hematological changes during ischemia/reperfusion and preconditioning in rats. METHODS: Forty healthy rats were divided into four groups: control, laparotomy, intestinal ischemia/reperfusion and ischemic preconditioning. The intestinal ischemia/reperfusion group received 45 min of superior mesenteric artery occlusion, while the ischemic preconditioning group received 10 min of short ischemia and reperfusion before 45 min of prolonged occlusion. A cell counter was used to analyze blood obtained from rats before and after the surgical procedures and the hematological results were compared among the groups. RESULTS: The results showed significant differences in hematimetric parameters among the groups. The parameters that showed significant differences included lymphocyte, white blood cells and granulocyte counts; hematocrit; mean corpuscular hemoglobin concentration; red cell deviation width; platelet count; mean platelet volume; plateletcrit and platelet distribution width. CONCLUSION: The most remarkable parameters were those related to leukocytes and platelets. Some of the data, including the lymphocyte and granulocytes counts, suggest that ischemic preconditioning attenuates the effect of intestinal ischemia/reperfusion on circulating blood cells. Our work contributes to a better understanding of the hematological responses after intestinal ischemia/reperfusion and IPC, and the present findings may also be used as predictive values. .

Continuous reinfusion of succus entericus associated with fistuloclysis in the management of a complex jejunal fistula on the abdominal wall.

INTRODUCTION: Fistuloclysis is an alternative method for enteral nutrition infusion, and has been successfully employed for the management of patients with high output small bowel fistula. However it has some deficiencies also. PRESENTATION OF CASE: A 42-year-old woman with multiple high output enterocutaneous fistula was submitted to fistuloclysis with reinfusion of chyme, after a period of several complications due to parenteral nutrition. DISCUSSION: Enteral nutrition provide better nutrition and fewer complications than parenteral nutrition. The enterocutaneous fistula usually does not allow enteral nutrition, however the use of fystuloclysis can fix this issue. The reinfusion of chyme provide the possibility of oral intake and better control of hydroeletrolitics disorders. CONCLUSION: More studies on the physiological effects of the chyme recirculation could add more data contributing to the clarification of this complex issue, but we believe that patients with high output and very proximal enterocutaneous fistula can be sucessfully treated with fistuloclysis and recirculation of chyme.

Effect of low-dose gaseous ozone on pathogenic bacteria.

BACKGROUND: Treatment of chronically infected wounds is a challenge, and bacterial environmental contamination is a growing issue in infection control. Ozone may have a role in these situations. The objective of this study was to determine whether a low dose of gaseous ozone/oxygen mixture eliminates pathogenic bacteria cultivated in Petri dishes. METHODS: A pilot study with 6 bacterial strains was made using different concentrations of ozone in an ozone-oxygen mixture to determine a minimally effective dose that completely eliminated bacterial growth. The small and apparently bactericidal gaseous dose of 20 µg/mL ozone/oxygen (1:99) mixture, applied for 5 min under atmospheric pressure was selected. In the 2nd phase, eight bacterial strains with well characterized resistance patterns were evaluated in vitro using agar-blood in adapted Petri dishes (105 bacteria/dish). The cultures were divided into 3 groups: 1--ozone-oxygen gaseous mixture containing 20 µg of O(3)/mL for 5 min; 2--100% oxygen for 5 min; 3--baseline: no gas was used. RESULTS: The selected ozone dose was applied to the following eight strains: Escherichia coli, oxacillin-resistant Staphylococcus aureus, oxacillin-susceptible Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, extended-spectrum beta-lactamase-producing Klebsiella pneumoniae, carbapenem-resistant Acinetobacter baumannii, Acinetobacter baumannii susceptible only to carbapenems, and Pseudomonas aeruginosa susceptible to imipenem and meropenem. All isolates were completely inhibited by the ozone-oxygen mixture while growth occurred in the other 2 groups. CONCLUSION: A single topical application by nebulization of a low ozone dose completely inhibited the growth of all potentially pathogenic bacterial strains with known resistance to antimicrobial agents.

Comparison of the neutrophil proteome in trauma patients and normal controls.

Neutrophils have an impressive array of microbicidal weapons, and in the presence of a pathogen, progress from a quiescent state in the bloodstream to a completely activated state. Failure to regulate this activation, for example, when the blood is flooded with cytokines after severe trauma, causes inappropriate neutrophil activation that paradoxically, is associated with tissue and organ damage. Acidic proteomic maps of quiescent human neutrophils were analyzed and compared to those of activated neutrophils from severe trauma patients. The analysis revealed 114 spots whose measured volumes differed between activated and quiescent neutrophils, with 27 upregulated and 87 downregulated in trauma conditions. Among the identified proteins, grancalcin, S100-A9 and CACNB2 reinforce observed correlations between motility and ion flux, ANXA3, SNAP, FGD1 and Zfyve19 are involved in vesicular transport and exocytosis, and GSTP1, HSPA1 HSPA1L, MAOB, UCH-L5, and PPA1 presented evidence that activated neutrophils may have diminished protection against oxidative damage and are prone to apoptosis. These are discussed, along with proteins involved in cytoskeleton reorganization, reactive oxygen species production, and ion flux. Proteins such as Zfyve19, MAOB and albumin- like protein were described for the first time in the neutrophil. In this work we achieved the identification of several proteins potentially involved in inflammatory signaling after trauma, as well as proteins described for the first time in neutrophils.

The effects of pneumoperitoneum and controlled ventilation on peritoneal lymphatic bacterial clearance: experimental results in rats.

OBJECTIVE: To evaluate the effect of pneumoperitoneum, both alone and in combination with controlled ventilation, on peritoneal lymphatic bacterial clearance using a rat bacterial peritonitis model. METHOD: A total of 69 male Wistar rats were intraperitoneally inoculated with an Escherichia coli solution (10(9) colony-forming units (cfu)/mL) and divided into three groups of 23 animals each: A (control group), B (pneumoperitoneum under 5 mmHg of constant pressure), and C (endotracheal intubation, controlled ventilation, and pneumoperitoneum as in Group B). The animals were sacrificed after 30 min under these conditions, and blood, mediastinal ganglia, lungs, peritoneum, liver, and spleen cultures were performed. RESULTS: Statistical analyses comparing the number of cfu/sample in each of the cultures showed that no differences existed between the three groups. CONCLUSION: Based on our results, we concluded that pneumoperitoneum, either alone or in association with mechanical ventilation, did not modify the bacterial clearance through the diaphragmatic lymphatic system of the peritoneal cavity.

The role of the New Trauma and Injury Severity Score (NTRISS) for survival prediction / O papel do New Trauma and Injury Severity Score (NTRISS) para predição de sobrevida / El papel del New Trauma and Injury Severity Score (NTRISS) para predicción de sobrevida

The objective of this study was to verify if replacing the Injury Severity Score (ISS) by the New Injury Severity Score (NISS) in the original Trauma and Injury Severity Score (TRISS) form would improve the survival rate estimation. This retrospective study was performed in a level I trauma center during one year. ROC curve was used to identify the best indicator (TRISS or NTRISS) for survival probability prediction. Participants were 533 victims, with a mean age of 38±16 years. There was predominance of motor vehicle accidents (61.9 percent). External injuries were more frequent (63.0 percent), followed by head/neck injuries (55.5 percent). Survival rate was 76.9 percent. There is predominance of ISS scores ranging from 9-15 (40.0 percent), and NISS scores ranging from 16-24 (25.5 percent). Survival probability equal to or greater than 75.0 percent was obtained for 83.4 percent of the victims according to TRISS, and for 78.4 percent according to NTRISS. The new version (NTRISS) is better than TRISS for survival prediction in trauma patients.

O objetivo deste estudo foi verificar se a substituição do Injury Severity Score (ISS) pelo New Injury Severity Score (NISS), na fórmula original do Trauma and Injury Severity Score (TRISS), melhora sua estimativa da taxa de sobrevida. Estudo retrospectivo realizado num centro de trauma nível I, durante um ano. A curva ROC foi utilizada para identificar o melhor indicador (TRISS ou NTRISS) para predição da probabilidade de sobrevida. O estudo incluiu 533 traumatizados, com idade média de 38±16 anos. Houve predomínio de acidentes de transporte (61,9 por cento). Lesões externas foram mais frequentes (63,0 por cento), seguidas por trauma craniencefálico/cervical (55,5 por cento). A taxa de sobrevida foi de 76,9 por cento. Houve predomínio dos valores do ISS variando de 9-15 (40,0 por cento) e, do NISS, de 16-24 (25,5 por cento). Probabilidade de sobrevida igual ou superior a 75,0 por cento foi obtida para 83,4 por cento das vítimas de acordo com o TRISS e por 78,4 por cento de acordo com NTRISS. A nova versão apresentou melhor desempenho que o TRISS na predição de sobrevida dos doentes traumatizados estudados.

El estudio objetivó verificar si la substitución del Injury Severity Score (ISS) por el New Injury Severity Score (NISS) en la fórmula original del Trauma and Injury Severity Score (TRISS) mejora su estimación de tasa de sobrevida. Estudio retrospectivo realizado en centro de trauma nivel I durante un año. Se usó curva ROC para identificar el mejor indicador (TRISS o NTRISS) para predicción de probabilidad de sobrevida. El estudio incluyó 533 traumatizados, edad media 38±16 años. Predominaron accidentes de transporte (61,9 por ciento). Prevalecieron las lesiones externas (63 por ciento), seguidas de trauma cráneo-encefálico/cervical (55,5 por ciento). Tasa de sobrevida: 76,9 por ciento. Predominaron valores de ISS variando de 9-15 (40,0 por ciento) y de NISS, de 16-24 (25,5 por ciento). Se obtuvo probabilidad de sobrevida igual o mayor a 75 por ciento para 83,4 por ciento de víctimas según TRISS y para 78,4 por ciento según NTRISS. La nueva versión (NTRISS) presentó mejor desempeño que TRISS en predicción de sobrevida en los pacientes traumatizados estudiados.

Diagnostic value of tomography of the cervical spine in victims of blunt trauma.

OBJECTIVE: to assess the value of computed tomography in the diagnosis of cervical spine and spinal cord injuries in victims of blunt trauma. METHODS: we reviewed the charts of blunt trauma victims from January 2006 to December 2008. We analyzed the following data: epidemiology, mechanism of trauma, transportation of victims to the hospital, intra-hospital care, indication criteria for CT, diagnosis, treatment and evolution of the victims. The victims were divided into two groups: Group I - without cervical spine injury, Group II - with cervical spine injury. RESULTS: we gathered medical records from 3,101 victims. Computed tomography was performed in 1572 (51%) patients, with male predominance (79%) and mean age of 38.53 years in Group I and 37.60 years in Group II. The distribution of trauma mechanisms was similar in both groups. Lesions found included: 53 fractures, eight vertebral listeses and eight spinal cord injuries. Sequelae included: paraplegia in three cases, quadriplegia in eight and brain injury in five. There were seven deaths in Group II and 240 in Group I. The average length of hospital stay was 11 days for Group I and 26.2 days for Group II. CONCLUSION. A CT scan of the cervical spine in victims of blunt trauma was effective in identifying lesions of the cervical spine and spinal cord injuries. Thus, despite the cost of neck CT and the low incidence of lesions identified by it, its indication based on the usual criteria seems justified.

Valor diagnóstico da tomografia de coluna cervical em vítimas de trauma contuso / Diagnostic value of tomography of the cervical spine in victims of blunt trauma

OBJETIVO. Avaliar o valor da tomografia computadorizada no diagnóstico de lesões da coluna e medula cervicais em vítimas de trauma contuso. MÉTODOS. Revisão dos prontuários de vítimas de trauma contuso atendidas de janeiro de 2006 a dezembro de 2008. Foram analisados os seguintes dados: epidemiológicos, mecanismo de trauma, transporte das vítimas para o hospital, atendimento intra-hospitalar, critérios de indicação da TC, diagnóstico, tratamento, e evolução das vítimas em estudo. As vítimas foram distribuídas em dois grupos: Grupo I - sem lesão na coluna cervical; Grupo II - com lesão na coluna cervical . RESULTADOS. Foram analisados os prontuários de 3.101 vítimas. A tomografia computadorizada foi indicada em 1.572 (51 por cento) pacientes, Foi observado predomínio masculino entre as vítimas (79 por cento), com média etária de 38,53 anos no Grupo I e 37,60 anos no Grupo II. A distribuição dos mecanismos de trauma foi semelhante nos dois grupos. Lesões encontradas: 53 fraturas, oito listeses vertebrais e oito lesões medulares. As sequelas incluíram: três paraplegias, cinco tetraplegias e oito sequelas de lesão cerebral. No Grupo II ocorreram sete óbitos ,no Grupo I 240. A duração média de internação hospitalar foi de 11 dias para o Grupo I e 26,2 dias para o Grupo II. CONCLUSÃO. A TC de coluna cervical em vítimas de trauma contuso foi eficaz na identificação de lesões da coluna e medula cervicais. Assim, apesar do custo da TC cervical, e da baixa incidência de lesões por ela identificáveis, a sua indicação baseada nos critérios usuais parece justificável.

OBJECTIVE: to assess the value of computed tomography in the diagnosis of cervical spine and spinal cord injuries in victims of blunt trauma. METHODS: we reviewed the charts of blunt trauma victims from January 2006 to December 2008. We analyzed the following data: epidemiology, mechanism of trauma, transportation of victims to the hospital, intra-hospital care, indication criteria for CT, diagnosis, treatment and evolution of the victims. The victims were divided into two groups: Group I - without cervical spine injury, Group II - with cervical spine injury. RESULTS: we gathered medical records from 3,101 victims. Computed tomography was performed in 1572 (51 percent) patients, with male predominance (79 percent) and mean age of 38.53 years in Group I and 37.60 years in Group II. The distribution of trauma mechanisms was similar in both groups. Lesions found included: 53 fractures, eight vertebral listeses and eight spinal cord injuries. Sequelae included: paraplegia in three cases, quadriplegia in eight and brain injury in five. There were seven deaths in Group II and 240 in Group I. The average length of hospital stay was 11 days for Group I and 26.2 days for Group II. CONCLUSION. A CT scan of the cervical spine in victims of blunt trauma was effective in identifying lesions of the cervical spine and spinal cord injuries. Thus, despite the cost of neck CT and the low incidence of lesions identified by it, its indication based on the usual criteria seems justified.

The role of the New Trauma and Injury Severity Score (NTRISS) for survival prediction.

The objective of this study was to verify if replacing the Injury Severity Score (ISS) by the New Injury Severity Score (NISS) in the original Trauma and Injury Severity Score (TRISS) form would improve the survival rate estimation. This retrospective study was performed in a level I trauma center during one year. ROC curve was used to identify the best indicator (TRISS or NTRISS) for survival probability prediction. Participants were 533 victims, with a mean age of 38±16 years. There was predominance of motor vehicle accidents (61.9%). External injuries were more frequent (63.0%), followed by head/neck injuries (55.5%). Survival rate was 76.9%. There is predominance of ISS scores ranging from 9-15 (40.0%), and NISS scores ranging from 16-24 (25.5%). Survival probability equal to or greater than 75.0% was obtained for 83.4% of the victims according to TRISS, and for 78.4% according to NTRISS. The new version (NTRISS) is better than TRISS for survival prediction in trauma patients.

The effects of pneumoperitoneum and controlled ventilation on peritoneal lymphatic bacterial clearance: experimental results in rats

OBJECTIVE: To evaluate the effect of pneumoperitoneum, both alone and in combination with controlled ventilation, on peritoneal lymphatic bacterial clearance using a rat bacterial peritonitis model. METHOD: A total of 69 male Wistar rats were intraperitoneally inoculated with an Escherichia coli solution (109 colony-forming units (cfu)/mL) and divided into three groups of 23 animals each: A (control group), B (pneumoperitoneum under 5 mmHg of constant pressure), and C (endotracheal intubation, controlled ventilation, and pneumoperitoneum as in Group B). The animals were sacrificed after 30 min under these conditions, and blood, mediastinal ganglia, lungs, peritoneum, liver, and spleen cultures were performed. RESULTS: Statistical analyses comparing the number of cfu/sample in each of the cultures showed that no differences existed between the three groups. CONCLUSION: Based on our results, we concluded that pneumoperitoneum, either alone or in association with mechanical ventilation, did not modify the bacterial clearance through the diaphragmatic lymphatic system of the peritoneal cavity.

Evaluation of the effects of ozone therapy in the treatment of intra-abdominal infection in rats.

INTRODUCTION: The antibacterial effect of ozone (O(3)) has been described in the extant literature, but the role of O(3) therapy in the treatment of certain types of infection remains controversial. OBJECTIVES: To evaluate the effect of intraperitoneal (i.p.) O(3) application in a cecal ligation/puncture rat model on interleukins (IL-6, IL-10) and cytokine-induced neutrophil chemoattractant (CINC)-1 serum levels, acute lung injury and survival rates. METHODS: FOUR ANIMAL GROUPS WERE USED FOR THE STUDY: a) the SHAM group underwent laparotomy; b) the cecal ligation/puncture group underwent cecal ligation/puncture procedures; and c) the CLP+O(2) and CLP+O(3) groups underwent CLP+ corresponding gas mixture infusions (i.p.) throughout the observation period. IL-6, CINC-1 and IL-10 concentrations were determined by enzyme-linked immunosorbent assay (ELISA). Acute lung injury was evaluated with the Evans blue dye lung leakage method and by lung histology. P<0.05 was considered significant. RESULTS: CINC-1 was at the lowest level in the SHAM group and was lower for the CLP+O(3) group vs. the CLP+O(2) group and the cecal ligation/puncture group. IL-10 was lower for the SHAM group vs. the other three groups, which were similar compared to each other. IL-6 was lower for the SHAM group vs. all other groups, was lower for the CLP+O(3) or CLP+O(2) group vs. the cecal ligation/puncture group, and was similar for the CLP+O(3) group vs. the CLP+O(2) group. The lung histology score was lower for the SHAM group vs. the other groups. The Evans blue dye result was lower for the CLP+O(3) group vs. the CLP+O(2) group and the cecal ligation/puncture group but similar to that of the SHAM group. The survival rate for the CLP+O(3) group was lower than for the SHAM group and similar to that for the other 2 groups (CLP and CLP+O(2)). CONCLUSION: Ozone therapy modulated the inflammatory response and acute lung injury in the cecal ligation/puncture infection model in rats, although there was no improvement on survival rates.

Evaluation of the effects of ozone therapy in the treatment of intra-abdominal infection in rats

INTRODUCTION: The antibacterial effect of ozone (O3) has been described in the extant literature, but the role of O3 therapy in the treatment of certain types of infection remains controversial. OBJECTIVES: To evaluate the effect of intraperitoneal (i.p.) O3 application in a cecal ligation/puncture rat model on interleukins (IL-6, IL-10) and cytokine-induced neutrophil chemoattractant (CINC)-1 serum levels, acute lung injury and survival rates. METHODS: Four animal groups were used for the study: a) the SHAM group underwent laparotomy; b) the cecal ligation/puncture group underwent cecal ligation/puncture procedures; and c) the CLP+O2 and CLP+O3 groups underwent CLP+ corresponding gas mixture infusions (i.p.) throughout the observation period. IL-6, CINC-1 and IL-10 concentrations were determined by enzyme-linked immunosorbent assay (ELISA). Acute lung injury was evaluated with the Evans blue dye lung leakage method and by lung histology. P<0.05 was considered significant. RESULTS: CINC-1 was at the lowest level in the SHAM group and was lower for the CLP+O3 group vs. the CLP+O2 group and the cecal ligation/puncture group. IL-10 was lower for the SHAM group vs. the other three groups, which were similar compared to each other. IL-6 was lower for the SHAM group vs. all other groups, was lower for the CLP+O3 or CLP+O2 group vs. the cecal ligation/puncture group, and was similar for the CLP+O3 group vs. the CLP+O2 group. The lung histology score was lower for the SHAM group vs. the other groups. The Evans blue dye result was lower for the CLP+O3 group vs. the CLP+O2 group and the cecal ligation/puncture group but similar to that of the SHAM group. The survival rate for the CLP+O3 group was lower than for the SHAM group and similar to that for the other 2 groups (CLP and CLP+O2). CONCLUSION: Ozone therapy modulated the inflammatory response and acute lung injury in the cecal ligation/puncture infection model in rats, although there was no ...

Avaliação de um esquema de realimentação utilizado após 43 dias de jejum voluntário / Evaluation of a refeeding plan employed after 43 days of voluntary fasting

Objetivo: Avaliar um esquema de realimentação utilizado em pacientes após jejum prolongado voluntário. Métodos: Foi realizado estudo retrospectivo, com base em levantamento dos prontuários de 7 pacientes (média de idade de 41,6 anos e desvio-padrão=5,3) em jejum voluntário por 43 dias em jejum seguidos de 14 dias de realimentação. As variáveis estudadas foram: antropométricas, clínicas, dietéticas, laboratoriais e avaliação da composição corpórea por bioimpedância. Foram utilizadas as seguintes dietas por via oral na realimentação: oligomérica enriquecida com glutamina, associada à dieta hipogordurosa e isenta de lactose (primeira semana); dieta branda normolipídica com lactose (segunda semana). A análise descritiva dos dados apresentados incluiu média e desvio-padrão. A análise de variância ou o teste não paramétrico de Kruskal-Wallis foramutilizados para comparação dos dados nos períodos de jejum e realimentação. Utilizou-se o programa estatístico SIGMA STAT versão 2,03 sendo considerado o nível de significância de 5%. Resultados: No jejum houve redução significativa do peso corpóreo (18,0% desvio-padrão=2,3), da prega cutânea do tríceps (48,2% desvio-padrão=9,0) e da hemoglobina (15,98g/dL desvio-padrão=0,99 para 12,74g/dL desvio-padrão=0,47). Na segunda semana de realimentação houve aumento significativo na contagem total de linfócitos (954 mil/mm3 desvio-padrão=242 para 1.619 mil/mm3 desvio-padrão=232) e dos distúrbios gastrointestinais, em relação à primeira semana. Não ocorreu Síndrome da Realimentação. Conclusão O esquema de realimentação utilizado promoveu resultado global adequado, mas requer redução do aporte energético e definição de momento mais adequado para reintrodução de lactose e da dieta normolipídica.

Objective: To evaluate a refeeding plan employed in patients after a long period of voluntary fasting. Methods: A retrospective study was done by reviewing the medical records of 7 male patients (mean age of 42.0 years; standard deviation of 5.3 years) who were refed for 14 days after a 43-day fast. The studied variables were:clinical, anthropometric; biochemical and body composition by bioimpedance analysis. The following diets were used orally in the refeeding program: glutamine-enriched oligomeric diet associated with a low-fat lactose-free diet (first week); normal-fat diet with lactose (second week). Descriptive analysis of the data included mean and standard deviation. Analysis of variance or the non-parametric Kruskal-Wallis test was used to compare data during the fasting and refeeding periods. The SIGMA STAT statistics software package version 2.03 was used to analyze data with the significance level set at 5% (p<0.05). Results: During the fasting period there was a significant reduction in body weight (18.0%; standard deviation=2.3), triceps skinfold (48.2%; standard deviation=9.0) and hemoglobin concentration (from 15.98g/dL; standard deviation=0.99 to 12.74g/dL; standard deviation=0.47). During the second week of refeeding, there was a significant increase in total lymphocyte count (from 954 thousand/mm3; standard deviation=242 to 1.619 thousand/mm3; standard deviation=232); and gastrointestinal problems when compared with the first week. No case of refeeding syndrome was observed.Conclusion The refeeding program used promoted a satisfactory global result but it is necessary to reduce energy intakeand define the best moment to reintroduce lactose and a normal-fat diet.

Gut ischemia/reperfusion induced acute lung injury is an alveolar macrophage dependent event.

BACKGROUND: Although the role of the lung alveolar macrophage (AM) as a mediator of acute lung injury (ALI) after lung ischemia/reperfusion (I/R) has been suggested by animal experiments, it has not been determined whether AMs mediate ALI after intestinal I/R. The objective of this study was to determine the effect of AM elimination on ALI after intestinal I/R in rats. METHODS: Male Wistar rats (n = 90) were randomly divided into three groups: the clodronate-liposomes (CLOD-LIP) group received intratracheal treatment with CLOD-LIP; the liposomes (LIP) group received intratracheal treatment with LIP; and the nontreated (UNTREAT) group received no treatment. Twenty-four hours later each group was randomly divided into three subgroups: the intestinal I/R subgroup was subjected to 45-minute intestinal ischemia and 2-hour reperfusion; the laparotomy (LAP) subgroup was subjected to LAP and sham procedures; the control (CTR) subgroup received no treatment. At the end of reperfusion, ALI was quantitated in all the animals by the Evans blue dye (EBD) method. RESULTS: ALI values are expressed as EBD lung leakage (microg EBD/g dry lung weight). EBD lung leakage values in the CLOD-LIP group were 32.59 +/- 12.74 for I/R, 27.74 +/- 7.99 for LAP, and 33.52 +/- 10.17 for CTR. In the LIP group, lung leakage values were 58.02 +/- 18.04 for I/R, 31.90 +/- 8.72 for LAP, and 27.17 +/- 11.48 for CTR. In the UNTREAT group, lung leakage values were 55.60 +/- 10.96 for I/R, 35.99 +/- 6.89 for LAP, and 30.83 +/- 8.41 for CTR. Within each group, LAP values did not differ from CTR values. However, in the LIP and UNTREAT groups, values for both the LAP and CTR subgroups were lower than values for the I/R subgroup (p < 0.001). The CLOD-LIP I/R subgroup value was less (p < 0.001) than the I/R subgroup values in the LIP and UNTREAT groups. These results indicated that I/R provokes ALI that can be prevented by CLOD-LIP treatment, and further suggested that AMs are essential for ALI occurrence induced by intestinal I/R in rats.