Influence of laparoscopy and laparotomy on gasometry, leukocytes and cytokines in a rat abdominal sepsis model

Laparoscopic surgery is associated with reduced surgical trauma, and less acute phase response, as compared with open surgery. Cytokines are important regulators of the biological response to surgical and anesthetic stress. The aim of this study was to determine if CO2 pneumoperitoneum would change cytokine expression, gas parameters and leukocyte count in septic rats. Methods: Wistar rats were randomly assigned to five groups: control (anesthesia only), laparotomy, CO2 pneumoperitoneum, cecum ligation and puncture by laparotomy, and laparoscopic cecum ligation and puncture. After 30 min of the procedures, arterial blood samples were obtained to determine leukocytes subpopulations by hemocytometer. TNFα, IL-1β, IL-6 were determined in intraperitoneal fluid (by ELISA). Gas parameters were measured on arterial blood, intraperitoneal and subperitoneal exsudates. Results: Peritoneal TNFα, IL-1β and IL-6 concentrations were lower in pneumoperitoneum rats than in all other groups (p<0.05). TNFα, IL-1β and IL-6 expression was lower in the laparoscopic than in laparotomic sepsis (p<0.05). Rats from laparoscopic cecum ligation and puncture group developed significant hypercarbic acidosis in blood and subperitoneal fluid when compared to open procedure group. Total white blood cells and lymphocytes were significantly lower in laparoscopic cecum ligation and puncture rats than in the laparotomic (p<0.01). Nevertheless, the laparotomic cecum ligation rats had a significant increase in blood neutrophils and eosinophils when compared with controls (p<0.05). Conclusions: This study demonstrates that the CO2 pneumoperitoneum reduced the inflammatory response in an animal model of peritonitis with respect to intraperitoneal cytokines, white blood cell count and clinical correlates of sepsis. The pneumoperitoneum produced hypercarbic acidosis in septic animals

Effects of simvastatin in abdominal sepsis in rats

Statins are widely recognized as hypolipemic drugs, but some studies have observed anti-inflammatory and immunomodulatory effects, known as pleiotropic. The aims of this work was to study possible anti-inflammatory effects of simvastatin in abdominal sepsis. Serum pro-inflammatory cytokines and leukocytes count were determined in an experimental model of abdominal sepsis, using cecal ligation and puncture (CLP) in rats. Methods: Twenty eigth Wistar rats weighing 285±12g were randomly divided in: CLP/Sinvastatin rats (n=7), treated with 10 mg/Kg of oral simvastatin 18 and 2 hs berofe CLP; CLP/Saline group rats (n=7), treated with oral saline; group Sham/Simvastatin (n=7), treated with simvastatin, and group Sham/Saline (n=7), treated with saline. Serum TNF-α, IL-1β and IL-6 by ELISA and total leukocytes, neutrophils, lymphocytes, and eosinophils were determined 24 hs after CLP. ANOVA and Tukey test were used considering significant p<0.05. Results: It was demonstrated that serum TNF-α, IL-1β and IL-6 were respectively 364,8±42pg/mL; 46,3±18pg/mL and 28,4±13pg/mL in CLP/Sinvastatin rats, significantly lower (p<0.05) than in group CLP/Saline (778,5±86pg/ml; 176,9±46pg/ ml; 133,6±21 pg/ml, respectively). The same results were observed in total leukocytes and neutrophils counts. Conclusion: These results clearly demonstrate that simvastatin is an effective agent that reduces cytokines levels and leukocyte count in sepsis, independently of its well-known lipid-lowering effects. Thus, HMG-CoA reductase inhibitors like simvastatin have important anti-inflammatory effects in abdominal sepsis in rats

Influence of laparoscopy and laparotomy on gasometry, leukocytes and cytokines in a rat abdominal sepsis model

Laparoscopic surgery is associated with reduced surgical trauma, and less acute phase response, as compared with open surgery. Cytokines are important regulators of the biological response to surgical and anesthetic stress. The aim of this study was to determine if CO2 pneumoperitoneum would change cytokine expression, gas parameters and leukocyte count in septic rats. Methods: Wistar rats were randomly assigned to five groups: control (anesthesia only), laparotomy, CO2 pneumoperitoneum, cecum ligation and puncture by laparotomy, and laparoscopic cecum ligation and puncture. After 30 min of the procedures, arterial blood samples were obtained to determine leukocytes subpopulations by hemocytometer. TNFα, IL-1β, IL-6 were determined in intraperitoneal fluid (by ELISA). Gas parameters were measured on arterial blood, intraperitoneal and subperitoneal exsudates. Results: Peritoneal TNFα, IL-1β and IL-6 concentrations were lower in pneumoperitoneum rats than in all other groups (p<0.05). TNFα, IL-1β and IL-6 expression was lower in the laparoscopic than in laparotomic sepsis (p<0.05). Rats from laparoscopic cecum ligation and puncture group developed significant hypercarbic acidosis in blood and subperitoneal fluid when compared to open procedure group. Total white blood cells and lymphocytes were significantly lower in laparoscopic cecum ligation and puncture rats than in the laparotomic (p<0.01). Nevertheless, the laparotomic cecum ligation rats had a significant increase in blood neutrophils and eosinophils when compared with controls (p<0.05). Conclusions: This study demonstrates that the CO2 pneumoperitoneum reduced the inflammatory response in an animal model of peritonitis with respect to intraperitoneal cytokines, white blood cell count and clinical correlates of sepsis. The pneumoperitoneum produced hypercarbic acidosis in septic animals

Effects of simvastatin in abdominal sepsis in rats

Statins are widely recognized as hypolipemic drugs, but some studies have observed anti-inflammatory and immunomodulatory effects, known as pleiotropic. The aims of this work was to study possible anti-inflammatory effects of simvastatin in abdominal sepsis. Serum pro-inflammatory cytokines and leukocytes count were determined in an experimental model of abdominal sepsis, using cecal ligation and puncture (CLP) in rats. Methods: Twenty eigth Wistar rats weighing 285±12g were randomly divided in: CLP/Sinvastatin rats (n=7), treated with 10 mg/Kg of oral simvastatin 18 and 2 hs berofe CLP; CLP/Saline group rats (n=7), treated with oral saline; group Sham/Simvastatin (n=7), treated with simvastatin, and group Sham/Saline (n=7), treated with saline. Serum TNF-α, IL-1β and IL-6 by ELISA and total leukocytes, neutrophils, lymphocytes, and eosinophils were determined 24 hs after CLP. ANOVA and Tukey test were used considering significant p<0.05. Results: It was demonstrated that serum TNF-α, IL-1β and IL-6 were respectively 364,8±42pg/mL; 46,3±18pg/mL and 28,4±13pg/mL in CLP/Sinvastatin rats, significantly lower (p<0.05) than in group CLP/Saline (778,5±86pg/ml; 176,9±46pg/ ml; 133,6±21 pg/ml, respectively). The same results were observed in total leukocytes and neutrophils counts. Conclusion: These results clearly demonstrate that simvastatin is an effective agent that reduces cytokines levels and leukocyte count in sepsis, independently of its well-known lipid-lowering effects. Thus, HMG-CoA reductase inhibitors like simvastatin have important anti-inflammatory effects in abdominal sepsis in rats

Efeito da sinvastatina na sepse abdominal de ratos diabéticos

Analisar se o pré-tratamento com sinvastatina em modelo experimental de sepse abdominal é benéfico em ratos diabéticos. Métodos: Cinquenta e seis ratos Wistar foram aleatoriamente distribuídos em: grupo não diabético (n-28) e grupo diabetes induzido por estreptozotocina (n=28). Sepse abdominal por ligadura e punção do ceco foi induzida em 14 ratos diabéticos e em 14 não diabéticos. Os demais 28 animais foram alocados em grupo sham. Os grupos de ratos com sepse e os sham (cada com sete animais) foram tratados com microemulsão oral de simvastatina (20 mg kg-1 day-1) e solução salina 0,9%, respectivamente. Sangue periférico foi usado para dosagem de TNFa, IL-1b, IL-6, proteína C reativa, procalcitonina, contagem de leucócitos e neutrófilos em todos os animais. A análise estatística foi realizada pela ANOVA e teste de Tukey, com p<0,05. Resultados: A sinvastatina reduziu a mortalidade nos ratos diabéticos. Os valores séricos de TNF-a, IL-1b, IL-6, proteína C reativa, procalcitonina, leucócitos e neutrófilos mostraram-se mais baixos nos ratos diabéticos e não diabéticos com sepse, tratados com sinvastatina, do que nos tratados com solução salina. Conclusão: A sinvastatina teve efeito antiinflamatório, que pode ter resultado em proteção contra a sepse em ratos diabéticos

Proteção pela glucana em modelo experimental de sepse

OBJETIVO: Neste trabalho avaliou-se a ação da glucana ? ? 1-3 insolúvel, um polissacarídeo extraído da parede celular interna do fungo Saccharomyces cerevisae, como agente imunoestimulante inespecífico em camundongos submetidos a modelo de sepse experimental. MÉTODOS: Foram utilizados 73 camundongos Swiss, os quais receberam glucana em diferentes doses pelas vias intraperitoneal (i.p.). Sepse difusa foi induzida através da técnica de ligadura e punção do ceco, transfixado e drenado com fio multifilamentar. RESULTADOS: Os animais tratados com glucana apresentaram um aumento significante no número de leucócitos no lavado peritoneal, diminuição no número de unidades formadoras de colônias bacterianas. Observou-se um aumento significante na sobrevida dos animais tratados com glucana insolúvel, a qual proporcionou um maior controle da infecção bacteriana por aumentar o número de células de defesa. CONCLUSÃO: A glucana insolúvel, quando usada em camundongos por via intraperitoneal, em modelo de sepse abdominal, contribuiu para melhorar a sobrevida, induziu proteção contra a formação de colônias bacterianas no líquido peritoneal e aumentou a migração leucocitária

Avaliação das atividades anti-inflamatória, anticoagulante e antiproliferativa do inibidor de quimotripsina das sementes de erythrina velutina (EvCI)

Studies indicate that several components were isolated from medicinal plants, which have antibacterial, antifungal, antitumor and anti-inflammatory properties. Sepsis is characterized by a systemic inflammation which leads to the production of inflammatory mediators exacerbated by excessive activation of inflammatory cells and disseminated intravascular coagulation (DIC), in which the human neutrophil elastase plays an important role in its pathogenesis. Several epidemiological studies suggest that components of plants, especially legumes, can play a beneficial role in reducing the incidence of different cancers. A chymotrypsin inhibitor of Kunitz (Varela, 2010) was purified from seeds of Erythrina velutina (Mulungu) by fractionation with ammonium sulfate, affinity chromatography on Trypsin-Sepharose, Chymotrypsin-Sepharose and ion exchange chromatography on Resource Q 1 ml (GE Healthcare) in system FPLC / AKTA. The inhibitor, called EvCI, had a molecular mass of 17 kDa determined by SDS-PAGE. The purified protein was able to inhibit human neutrophil elastase (HNE), with an IC50 of 3.12 nM. The EvCI was able to inhibit both pathways of HNE release stimulated by PAF and fMLP (75.6% and 65% respectively). The inhibitor also inhibited leukocyte migration in septic mice about 87% and prolonged the time of coagulation and inhibition factor Xa. EvCI showed neither hemolytic activity nor cytotoxicity. EvCI showed a selective antiproliferative effect to HepG2 cell lines with IC50 of 0.5 micrograms per milliliter. These results suggest EvCI as a molecule antagonist of PAF / fMLP and a potential use in fighting inflammation related disorders, disseminated intravascular coagulation (DIC) and cancer

Um novo inibidor de tripsina multifuncional de sementes de Erythrina velutina: caracterizacao e propriedades farmacologicas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Influência da laparoscopia e da laparotomia na gasometria, leucometria diferencial e citocinas em modelo de sepse abdominal em ratos

Laparoscopic surgery is associated with reduced surgical trauma, and less acute phase response, as compared with open surgery. Cytokines are important regulators of the biological response to surgical and anesthetic stress. The aim of this study was to determine if CO2 pneumoperitoneum would change cytokine expression, gas parameters and leukocyte count in septic rats. Methods: Wistar rats were randomly assigned to five groups: control (anesthesia only), laparotomy, CO2 pneumoperitoneum, cecum ligation and puncture by laparotomy, and laparoscopic cecum ligation and puncture. After 30 min of the procedures, arterial blood samples were obtained to determine leukocytes subpopulations by hemocytometer. TNFα, IL-1β, IL-6 were determined in intraperitoneal fluid (by ELISA). Gas parameters were measured on arterial blood, intraperitoneal and subperitoneal exsudates. Results: Peritoneal TNFα, IL-1β and IL-6 concentrations were lower in pneumoperitoneum rats than in all other groups (p<0.05). TNFα, IL-1β and IL-6 expression was lower in the laparoscopic than in laparotomic sepsis (p<0.05). Rats from laparoscopic cecum ligation and puncture group developed significant hypercarbic acidosis in blood and subperitoneal fluid when compared to open procedure group. Total white blood cells and lymphocytes were significantly lower in laparoscopic cecum ligation and puncture rats than in the laparotomic (p<0.01). Nevertheless, the laparotomic cecum ligation rats had a significant increase in blood neutrophils and eosinophils when compared with controls (p<0.05). Conclusions: This study demonstrates that the CO2 pneumoperitoneum reduced the inflammatory and immune response in an animal model of peritonitis with respect to intraperitoneal cytokines, white blood cell count and clinical correlates of sepsis. The pneumoperitoneum produced hypercarbic acidosis in septic animals

Ação da β-1,3 glucana no sistema imune de camundongos com sepse por Candida albicans

The frequency of disseminated candidiasis caused by yeast has enhancing in intensive care unit. Despite the availability of new antifungal drugs, C. albicans sepsis mortality causes can be as high as 30-40%. So, it has been needed to looking for a new therapeutic medicament that helps in treatment and prevention of this infection. Previous data that demonstrated that particulated β-glucan stimulates the immune system and experiments of this work were conducted to investigating if β-glucan extracted from Saccharomices cerevisiae, could modified the evolution of mouse model C. albicans systemic infection. Balb/c mice with sepsis and β-1,3 glucan treated or not were analyzed the influence of β-1,3 glucan in survival of the animals, in the fungal burdens in kidney, in the production of urea and TNF even in the histopathology of kidney. The experiments shown that the infected animals a nd glucan treated had great survival (p<0,05), less unit form colony in kidney and normal levels of urea. In the kidney histopathology of not glucan treated animals it has seen more lesions when compared with treated animals. So we conclude that β-1,3 glucan could stimulate the immune system against disseminated C. albicans

Assistência de enfermagem e o risco para úlceras de pressão medido pela Escala de Braden em unidade de terapia intensiva

The problem facing the incidence of pressure ulcers (UP) in the hospital environment especially in the intensive care unit (ICU), although it is an old and frequent event in our professional practice, it is not notified in the researches as much as it should be. We observed a tendency to invest in therapeutical and in studies about the production of sophisticated new bandages. Few, however, are the investments in research on preventive measures in order to prevent or at least slow down the development of lesions. In this sense, the study aimed to analyze the correlation between nursing care and the risk of developing UP measured by the Braden scale in ICU patients. This is a descriptive study of longitudinal quantitative approach. The project obtained a favourable opinion from the Ethics Committee of HUOL (no 486/10). Data collection was carried out in the Hospital of Unimed in Natal during six months in 2011. The sample was of 32 patients hospitalized in ICU for over four days. The results were processed in SPSS 15.0 for descriptive statistics and inferential statistics. We identified that, only 9.4% of our sample developed UP, being predominantly male, elderly people aged above 60 years, Caucasian, with diagnostic hypothesis at the time of hospitalization of sepsis, were clinical patients, who presented hemodynamic instability, using orotracheal tube (TOT), enteral probe (SNE), vesical probe delay (SVD) and had values of albumin and hemoglobin levels below normal. In addition, these patients had a longer hospital stay, longer usage of TOT, SNE, SVD, increased use of sedation and drain than those who did not develop UP and were all at risk for developing these injuries second Braden scores. 66.7% of the lesions developed were located in the sacral region, limiting the degree I and all patients that developed were considered serious, 100.0% of them have evolved since the death. Small were the differences between the averages of Braden scores between patients with and without UP, 11,9+2,4 against 12,4+2,6 with p = 0.627. The clinical aspects of the patients in the study were instrumental in the development of UP, once, these findings were statistically significant through the Mann-Whitney test, and appropriateness of nursing conduct was decisive for the prevention of pressure ulcers in critical patients, since many were those classified as at risk (28) and few who have developed lesions (03)

Sinalização endógena do sistema Nociceptina/Orfanina FQ - receptor NOP: envolvimento na modulação da depressão experimental induzida por lipopolissacarídeo

During the last decades, it has been established that there is a relationship between major depression and activation of immune system. Nociceptin/orphanin FQ (N/OFQ) is the natural ligand of a Gi-protein coupled receptor named NOP, both compose the peptidergic system wich is involved in the regulation of mood states and inflammatory responses. Considering these actions, the present thesis aimed to investigate the consequences of blocking NOP signaling in lipopolysaccharide (LPS)-induced sickness and depressive-like behaviors in mice. Systemic administration of LPS doses, that do not cause sepsis in mice, induce changes in their behaviors related with activity of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukins 6 (IL-6) and 1β (IL-1 β). At the time points of 2 to 6 h and 24 h after intraperitoneal injection, mice treated with LPS displayed, respectively, sickness and depressive-like behaviors. In the present work the administration of LPS 0.8 mg/kg (ip) significantly induced sickness signs in Swiss and CD-1 mice, such as weight loss, transient reduction in rectal temperature and decrease of food and water intake. Moreover at 24 h after LPS injection these same mice strains displayed significantly increased immobility time on the tail suspension test (TST) when compared with control mice, this alteration was not related with possible locomotion impairments as verified on the open field test. Treatment with Nortriptyline 30 mg/kg (ip, 60 min prior the TST) reduced the immobility time of control and LPS-treated mice and was used as standard antidepressant. The NOP receptor antagonist SB-612111 (10 mg/kg, ip), 30 min prior LPS, did not modify LPS-induced sickness signs and depressive-like behavior. However, when injected 24 h after LPS treatment, SB-612111 (ip, 30 min prior the TST) as well as the peptidergic NOP receptor antagonist UFP-101 (10 nmol/2μL, icv, 5 min prior the TST) significantly reversed the toxin effects. The protocol of LPS-induced depressive-like states was also tested in NOP receptor knockout mice (NOP(-/-)) and their respective wild types (NOP(+/+)). LPS evoked transient rectal temperature reduction in NOP(-/-) mice and loss of body weight, food and water intake reduction in both NOP(+/+) and NOP(-/-) mice. The consumption of water was significantly different due to the genotype. LPS injection induced transient changes in pro-inflammatory cytokines. At 6 h after LPS injection, serum levels of TNF-α were significantly increased in NOP(+/+) and NOP(-/-) mice, as the IL-6 levels were significantly increased just in NOP(+/+) serum. At 24 h after LPS treatment the pro-inflammatory cytokines had returned to the baseline levels in both genotypes. LPS treatment elicited depressive-like effects in NOP(+/+) but not in NOP(-/-) mice. The data obtained during the execution of this doctoral thesis reveal that pharmacological and genetic blockade of NOP signaling does not affect LPS evoked sickness signs while reversing depressive-like behavior. In conclusion, these results highlight the involvement of the peptidergic system N/OFQ - NOP receptor in the modulation of behaviors related to mood and activation of the immune system.

Gasto com a internação de idosos em unidades de terapia intensiva

Introduction: Population aging in Brazil underscores the need to discuss the proper management of the budget allocated in health field, especially in the sectors of high complexity, where coexist costly procedures, limited resources and the need for cost containment. In the other hand, demand is growing in a way directly proportional to the increase in the number of elderly in country. Objective: In this way, this research had as main objective to analyze the costs resulting from the admission of elderly in intensive care units (ICU) and its associated factors. Methods: This is a cross-sectional study with a quantitative approach and featured as a descriptive and exploratory research. Data were collected from medical records of elderly hospitalized in ICU from a brazilian city called Natal-RN, between november first, 2013 and january, 31 of 2014. The variables collected relate to the socio demographic profile, morbidity framework and characterization of hospitalization. The dependent variable was categorized by quartile 75 in high and low expense of hospitalization and submitted to chi-square test with the independent variables of the survey. Associations with p value <0.20 in the bivariate analysis were submitted to the technique of multiple logistic regression. We opted for the construction of three regression models from the above algorithm: general regression model, composed by all 493 hospitalizations in the study, other made with 181 individuals admitted in health public system (SUS) and a third one related to 312 cases from private service in health area. Results: In the general regression model, the variables respiratory diseases, hospitalizations in the private system, disoriented patient and previous stroke were associated with greater probability of high spending in the ICU. In the other hand, in SUS kind of hospitalizations, this probability was associated with disoriented patient, 80 years old or more, sepsis and admission for clinical reason. In the cases from the private network health, the high expenditure was associated with respiratory disease, mechanical ventilation, hospitalization for clinical reason and disoriented patients. Conclusion: The increased expenditure on hospitalization of elderly in intensive care depends on the clinical conditions of individuals. This highlights the importance of avoiding hospitalizations due to diseases sensitive to primary care by health preventive actions and providing comprehensive care to the elderly. In addition, obtaining different explanatory models, according to kind hospital funding, demonstrates the importance of the organization in health services related to composition of costs of hospitalization among the elderly. Another question founded was the need that to improve the funding, we must use rationally the available resources by avoiding unnecessary hospitalizations of elderly people in the extremes of severity. On this kind of precarious funding, ICU hospitalization of elderly non-critical or in a terminal state can compromise the quality of services provided to those who really need intensive care.

Protocolo para assistência do enfermeiro ao paciente séptico em terapia intensiva: construção e validação de conteúdo

Tratar-se-á de um estudo metodológico, com abordagem quantitativa; baseado no referencial metodológico da técnica Delphi, com objetivo de construir e validar um protocolo clínico para assistência do enfermeiro ao paciente séptico na Unidade de Terapia Intensiva. A proposta metodológica seguiu duas etapas: elaboração do instrumento por meio da revisão integrativa da literatura e validação de conteúdo do protocolo. O estudo foi aprovado pelo Comitê de Ética em Pesquisa, mediante o Parecer CAAE 41873314.5.0000.5537. O universo amostral foi composto por profissionais enfermeiros, considerados peritos, doutores ou mestres na área da saúde, com experiência em alta complexidade e/ ou estudos de validação de instrumento/protocolo, selecionados por meio da Plataforma Lattes. Referente à primeira etapa elaborou-se um instrumento composto pela caracterização profissional dos peritos; e baseado em evidência científica e nas diretrizes do Surviving Sepsis Campaign, contemplando três tópicos assistenciais ao paciente com sepse, a saber: Triagem para Sepse- Reconhecimento das Manifestações Clínicas; Pacote de Ressuscitação Inicial (Controle das Primeiras 6 Horas); Tratamento de Suporte. A segunda etapa caracterizou-se na validação de conteúdo do instrumento para elaboração final do protocolo, utilizando à técnica Delphi, em duas fases. No que concernem as variáveis referentes ao estudo, na 1ª fase de Delphi, 34 peritos avaliaram o instrumento composto por 18 itens, no período de maio a julho de 2015, e os dados foram analisados pela estatística descritiva (frequência, média, mediana e desvio padrão) e pelo Índice de Validade de Conteúdo (IVC), demonstrando um IVC extremamente satisfatório para 15 itens, com total de 0,79, obtendo assim, a reformulação e refinamento do conteúdo do instrumento. Na segunda fase de Delphi, entre julho e agosto de 2015, participaram 26 peritos, e utilizou-se o percentual de concordância acima de 75% para as variáveis consideradas pertinentes ao protocolo de cuidados ao paciente séptico em UTI, obtendo nesta fase, o percentual de concordância de 95%. O protocolo foi concluído com 15 itens, sendo respaldado e modificado, baseado em evidência científica, nas diretrizes internacionais e nas sugestões dos peritos. A utilização do protocolo proposto poderá contribuir para a prática clínica do enfermeiro ao paciente séptico na Unidade de Terapia Intensiva.

Protocolo para assistência do enfermeiro ao paciente séptico em terapia intensiva: construção e validação de conteúdo

Tratar-se-á de um estudo metodológico, com abordagem quantitativa; baseado no referencial metodológico da técnica Delphi, com objetivo de construir e validar um protocolo clínico para assistência do enfermeiro ao paciente séptico na Unidade de Terapia Intensiva. A proposta metodológica seguiu duas etapas: elaboração do instrumento por meio da revisão integrativa da literatura e validação de conteúdo do protocolo. O estudo foi aprovado pelo Comitê de Ética em Pesquisa, mediante o Parecer CAAE 41873314.5.0000.5537. O universo amostral foi composto por profissionais enfermeiros, considerados peritos, doutores ou mestres na área da saúde, com experiência em alta complexidade e/ ou estudos de validação de instrumento/protocolo, selecionados por meio da Plataforma Lattes. Referente à primeira etapa elaborou-se um instrumento composto pela caracterização profissional dos peritos; e baseado em evidência científica e nas diretrizes do Surviving Sepsis Campaign, contemplando três tópicos assistenciais ao paciente com sepse, a saber: Triagem para Sepse- Reconhecimento das Manifestações Clínicas; Pacote de Ressuscitação Inicial (Controle das Primeiras 6 Horas); Tratamento de Suporte. A segunda etapa caracterizou-se na validação de conteúdo do instrumento para elaboração final do protocolo, utilizando à técnica Delphi, em duas fases. No que concernem as variáveis referentes ao estudo, na 1ª fase de Delphi, 34 peritos avaliaram o instrumento composto por 18 itens, no período de maio a julho de 2015, e os dados foram analisados pela estatística descritiva (frequência, média, mediana e desvio padrão) e pelo Índice de Validade de Conteúdo (IVC), demonstrando um IVC extremamente satisfatório para 15 itens, com total de 0,79, obtendo assim, a reformulação e refinamento do conteúdo do instrumento. Na segunda fase de Delphi, entre julho e agosto de 2015, participaram 26 peritos, e utilizou-se o percentual de concordância acima de 75% para as variáveis consideradas pertinentes ao protocolo de cuidados ao paciente séptico em UTI, obtendo nesta fase, o percentual de concordância de 95%. O protocolo foi concluído com 15 itens, sendo respaldado e modificado, baseado em evidência científica, nas diretrizes internacionais e nas sugestões dos peritos. A utilização do protocolo proposto poderá contribuir para a prática clínica do enfermeiro ao paciente séptico na Unidade de Terapia Intensiva.