Effects of exosomes from human adipose-derived mesenchymal stem cells on pulmonary vascular endothelial cells injury in septic mice and its mechanism / 中华烧伤杂志

Objective: To investigate the effects of exosomes from human adipose-derived mesenchymal stem cells (ADSCs) on pulmonary vascular endothelial cells (PMVECs) injury in septic mice and its mechanism. Methods: The experimental research method was adopted. The primary ADSCs were isolated and cultured from the discarded fresh adipose tissue of 3 patients (female, 10-25 years old), who were admitted to the First Affiliated Hospital of Air Force Medical University undergoing abdominal surgery, and the cell morphology was observed by inverted phase contrast microscope on the 5th day. The expressions of CD29, CD34, CD44, CD45, CD73, and CD90 of ADSCs in the third passage were detected by flow cytometry. The third to the fifth passage of ADSCs were collected, and their exosomes from the cell supernatant were obtained by differential ultracentrifugation, and the shape, particle size, and the protein expressions of CD9, CD63, tumor susceptibility gene 101 (TSG101), and β-actin of exosomes were detected, respectively, by transmission electron microscopy, nano-particle tracking analysis and Western blotting. Twenty-four adult male BALB/c mice were adopted and were divided into normal control group, caecal ligation perforation (CLP) alone group, and CLP+ADSC-exosome group with each group of 8 according to random number table (the same grouping method below) and were treated accordingly. At 24 h after operation, tumor necrosis factor (TNF-α) and interleukin 1β (IL-1β) levels of mice serum were detected by enzyme-linked immunosorbent assay, and lung tissue morphology of mice was detected by hematoxylin-eosin and myeloperoxidase staining, and the expression of 8-hydroxy-deoxyguanosine (8-OHdG) of mouse lung cells was detected by immunofluorescence method. Primary PMVECs were obtained from 1-month-old C57 mice regardless gender by tissue block method. The expression of CD31 of PMVECs was detected by immunofluorescence and flow cytometry. The third passage of PMVECs was co-cultured with ADSCs derived exosomes for 12 h, and the phagocytosis of exosomes by PMVECs was detected by PKH26 kit. The third passage of PMVECs were adopted and were divided into blank control group, macrophage supernatant alone group, and macrophage supernatant+ADSC-exosome group, with 3 wells in each group, which were treated accordingly. After 24 h, the content of reactive oxygen species in cells was detected by flow cytometry, the expression of 8-OHdG in cells was detected by immunofluorescence, and Transwell assay was used to determine the permeability of cell monolayer. The number of samples in above were all 3. Data were statistically analyzed with one-way analysis of variance and least significant difference t test. Results: The primary ADSCs were isolated and cultured to day 5, growing densely in a spindle shape with a typical swirl-like. The percentages of CD29, CD44, CD73 and CD90 positive cells of ADSCs in the third passage were all >90%, and the percentages of CD34 and CD45 positive cells were <5%. Exosomes derived from ADSCs of the third to fifth passages showed a typical double-cavity disc-like structure with an average particle size of 103 nm, and the protein expressions of CD9, CD63 and TSG101 of exosomes were positive, while the protein expression of β-actin of exosomes was negative. At 24 h after operation, compared with those in normal control group, both the levels of TNF-α and IL-1β of mice serum in CLP alone group were significantly increased (with t values of 28.76 and 29.69, respectively, P<0.01); compared with those in CLP alone group, both the content of TNF-α and IL-1β of mice serum in CLP+ADSC-exosome group was significantly decreased (with t values of 9.90 and 4.76, respectively, P<0.05 or P<0.01). At 24 h after surgery, the pulmonary tissue structure of mice in normal control group was clear and complete without inflammatory cell infiltration; compared with those in normal control group, the pulmonary tissue edema and inflammatory cell infiltration of mice in CLP alone group were more obvious; compared with those in CLP alone group, the pulmonary tissue edema and inflammatory cell infiltration of mice in CLP+ADSC-exosome group were significantly reduced. At 24 h after operation, endothelial cells in lung tissues of mice in 3 groups showed positive expression of CD31; compared with that in normal control group, the fluorescence intensity of 8-OHdG positive cells of the lung tissues of mice in CLP alone group was significantly increased, and compared with that in CLP alone group, the fluorescence intensity of 8-OHdG positive cells in the lung tissues of mice in CLP+ADSC-exosome group was significantly decreased. The PMVECs in the 3rd passage showed CD31 positive expression by immunofluorescence, and the result of flow cytometry showed that CD31 positive cells accounted for 99.5%. At 12 h after co-culture, ADSC-derived exosomes were successfully phagocytose by PMVECs and entered its cytoplasm. At 12 h after culture of the third passage of PMVECs, compared with that in blank control group, the fluorescence intensity of reactive oxygen species of PMVECs in macrophage supernatant alone group was significantly increased (t=15.73, P<0.01); compared with that in macrophage supernatant alone group, the fluorescence intensity of reactive oxygen species of PMVECs in macrophage supernatant+ADSC-exosome group was significantly decreased (t=4.72, P<0.01). At 12 h after culture of the third passage of PMVECs, and the 8-OHdG positive fluorescence intensity of PMVECs in macrophage supernatant alone group was significantly increased; and compared with that in blank control group, the 8-OHdG positive fluorescence intensity of PMVECs in macrophage+ADSC-exosome supernatant group was between blank control group and macrophage supernatant alone group. At 12 h after culture of the third passage PMVECs, compared with that in blank control group, the permeability of PMVECs monolayer in macrophage supernatant alone group was significantly increased (t=6.34, P<0.01); compared with that in macrophage supernatant alone group, the permeability of PMVECs monolayer cells in macrophage supernatant+ADSC-exosome group was significantly decreased (t=2.93, P<0.05). Conclusions: Exosomes derived from ADSCs can ameliorate oxidative damage in mouse lung tissue, decrease the level of reactive oxygen species, 8-OHdG expression, and permeability of PMVECs induced by macrophage supernatant.

Suppressive activity of RGX-365 on HMGB1-mediated septic responses

Abstract RGX-365 is the main fraction of black ginseng conmprising protopanaxatriol (PPT)-type rare ginsenosides (ginsenosides Rg4, Rg6, Rh4, Rh1, and Rg2). No studies on the antiseptic activity of RGX-365 have been reported. High mobility group box 1 (HMGB1) is recognized as a late mediator of sepsis, and the inhibition of HMGB1 release and recovery of vascular barrier integrity have emerged as attractive therapeutic strategies for the management of sepsis. In this study, we examined the effects of RGX-365 on HMGB1-mediated septic responses and survival rate in a mouse sepsis model. RGX-365 was administered to the mice after HMGB1 challenge. The antiseptic activity of RGX-365 was assessed based on the production of HMGB1, measurement of permeability, and septic mouse mortality using a cecal ligation and puncture (CLP)-induced sepsis mouse model and HMGB1-activated human umbilical vein endothelial cells (HUVECs). We found that RGX-365 significantly reduced HMGB1 release from LPS- activated HUVECs and CLP-induced release of HMGB1 in mice. RGX-365 also restored HMGB1-mediated vascular disruption and inhibited hyperpermeability in the mice. In addition, treatment with RGX-365 reduced sepsis-related mortality in vivo. Our results suggest that RGX- 365 reduces HMGB1 release and septic mortality in vivo, indicating that it is useful in the treatment of sepsis.

Coagulase-negative staphylococci isolates from blood cultures of newborns in a tertiary hospital in southern Brazil

Abstract Neonatal sepsis continues to be a major cause of morbidity and mortality worldwide. Coagulase-negative staphylococci (CoNS), commonly found on the skin, being the main agents isolated. The aim of this study was to evaluate CoNS isolated from blood cultures of newborn (NB) infants. The study took place between 2014 and 2016/2017 in a tertiary hospital in southern Brazil. Using the VITEK 2 system (bioMérieux, Marcy l'Etoile, France), the microorganisms were identified and had their sensitivity profiles determined. The minimum inhibitory concentrations of linezolid, tigecycline, and vancomycin were also determined. The clinical parameters and mortality rates of NBs were evaluated. From January to December 2014, 176 CoNS isolates were obtained from 131 patients and from June 2016 to July 2017, 120 CoNS isolates were obtained from 79 patients. Staphylococcus epidermidis was most prevalent in both periods. Resistance rates increased between 2014 and 2016/2017, especially against ciprofloxacin (52.27% and 73.11%, p = 0.0004), erythromycin (51.40% and 68.07%, p = 0.0054), gentamicin (50.59% and 67.23%, p = 0.0052), and penicillin (71.3% and 99.17%, p = 0.0001), respectively. With 100% susceptibility to linezolid, tigecycline, and vancomycin in both periods and methodologies tested. In 2014, 53.44% of the NBs received antibiotic therapy, and of these, 77.14% used a catheter; in 2016/2017, these were 78.48% and 95.16%, respectively. Regarding laboratory tests, a hemogram was ineffective, since patients with sepsis presented normal reference values. In 2014 and 2016/17, 15.71% and 17.74% of the NBs died, respectively. S. epidermidis was the predominant microorganism, related to catheter use in most cases. The resistance rates have increased over time, demonstrating the importance of adopting control and prevention measures in this hospital. CoNS are responsible for a significant neonatal sepsis mortality rate in infants.

Diagnostic accuracy of 18F-FDG-PET in abdominal sepsis in rats

Abstract Purpose The objective of this study was to investigate the accuracy of 18F-FDG-PET in the diagnosis of multibacterial abdominal sepsis by cecum ligation and puncture (CLP) in rats. Methods Adult Wistar rats ( Rattus norvegicus ), weighing 227±35g, were allocated into a sepsis group by CLP (n=10) and sham group (n=10). 18F-FDG-PET using microPET was performed on all rats after 24 hours. Results All animals survived for postoperative 24h. The abdomen/liver ratio of the standardized uptake value (SUV) percentage was significantly higher in the sepsis group than in the sham (p=0.004). The ROC curve showed an accuracy of 18F-FDG-PET to detect abdominal sepsis of 88.9% (p=0.001), sensitivity of 90% and specificity of 88.9%. When a cut-off point of 79% of the ratio between the SUV on the abdominal region and liver was established, the sensitivity was 90%, specificity of 88.9%; positive and negative predictive values of 90.0% and 88.9%, respectively. Conclusions The diagnostic accuracy of 18F-FDG-PET in rats with abdominal sepsis was significantly high. It was also demonstrated the predictive ability of the abdomen/liver SUV ratio to diagnose abdominal sepsis. These findings may have implications for the clinical setting, locating septic foci with PETscan.

Evaluation of the effect of intraperitoneal etanercept administration on oxidative stress and inflammation indicators in the kidney and blood of experimental sepsis-induced rats

Abstract INTRODUCTION: Sepsis is an important cause of mortality and morbidity, and inflammatory response and oxidative stress play major roles underlying its pathophysiology. Here, we evaluated the effect of intraperitoneal etanercept administration on oxidative stress and inflammation indicators in the kidney and blood of experimental sepsis-induced rats. METHODS: Twenty-eight adult Sprague Dawley rats were classified into Control (Group 1), Sepsis (Group 2), Sepsis+Cefazolin (Group 3), and Sepsis+Cefazolin+Etanercept (Group 4) groups. Kidney tissue and serum samples were obtained for biochemical and histopathological investigations and examined for the C reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), triggering receptor expressed on myeloid cells (TREM), and malondialdehyde (MDA) levels. RESULTS: The levels of TNF-α, TREM, and MDA in serum and kidney samples were significantly higher in rats from sepsis group than in rats from control group (p < 0.05). Group 3 showed a significant reduction in serum levels of TNF-α, CRP, and TREM as compared with Group 2 (p < 0.05). Serum TNF-α, CRP, TREM, and MDA levels and kidney TNF-α and TREM levels were significantly lower in Group 4 than in Group 2 (p < 0.05). Serum TNF-α and TREM levels in Group 4 were significantly lower than those in Group 3, and histopathological scores were significantly lower in Group 3 and Group 4 than in Group 2 (p < 0.05). Histopathological scores of Group 4 were significantly lower than those of Group 3 (p < 0.05). CONCLUSIONS: Etanercept, a TNF-α inhibitor, may ameliorate sepsis-induced oxidative stress, inflammation, and histopathological damage.

Effects of intraperitoneal glutamine in the treatment of experimental sepsis / Efeitos da glutamina intraperitoneal no tratamento da sepse experimental

ABSTRACT Background: Sepsis is an important public health issue and is associated with high treatment costs and high mortality rates. Glutamine supplementation has proven to be beneficial to the functions of the immune system, acting beneficially in the evolution of patients in severe catabolic states. Aim: To evaluate the effect of glutamine supplementation via intraperitoneal in rats, induced sepsis, considering the following organs: intestines, liver, kidneys and lungs. Methods: Male Wistar rats subjected to sepsis by ligature and cecal puncture were divided into two groups: control C (n=6) and glutamine G (n=11), in which were administered dipeptiven 20% at a dose of 2 ml/kg/day (equivalent to 0.4g N(2)-L-alanyl-L-glutamine/kg) intraperitoneally 48 h prior to sepsis induction. After 48 h they were euthanized and intestine, liver, lung and kidney were removed for histological analysis. Results: Intestinal epithelial desquamation of the control group was more intense compared to the glutamine group (p=0.008). In the kidneys, degenerative tubular epithelial changes were less severe in the animals that received glutamine (p=0.029). Regarding to the liver, glutamine group showed lower levels of cell swelling than the control group (p=0.034). In the lung there were no results with statistical significance. Conclusion: Prior intraperitoneal supplementation with glutamine in experimental animals is able to reduce the damage to the intestinal mucosa, to the kidneys and liver's histoarchitecture.

RESUMO Racional: A sepse é importante problema de saúde pública, sendo relacionada com altos custos de tratamento e elevadas taxas de mortalidade. A suplementação de glutamina tem provado ser benéfica às funções do sistema imune, atuando em estados catabólicos graves. Objetivo: Avaliar o efeito da suplementação de glutamina via intraperitoneal em ratos induzidos à sepse. Método: Foram utilizados ratos Wistar submetidos à sepse por ligadura e punção do ceco, separados em grupo controle C (n=6) e glutamina G (n=11), aos quais foram administrados dipeptiven a 20% com dose de 2 ml/kg/dia (equivalente a 0,4 g N(2)-L-alanil-L-glutamina/kg), via intraperitoneal, 48 h antes da indução da sepse. Após 48 h todos os animais foram submetidos à eutanásia e intestino, fígado, pulmão e rim foram retirados para análise histológica. Resultados: No intestino a descamação epitelial do grupo controle foi mais intensa em comparação ao da glutamina (p=0,008). Nos rins, houve menor degeneração do epitélio tubular nos animais que receberam glutamina (p=0,029). No fígado, o grupo glutamina apresentou índices menores de tumefação celular do que o grupo controle (p=0,034). No pulmão não houve resultados com significância estatística. Conclusão: A suplementação prévia de animais experimentais com glutamina via intraperitoneal é capaz de reduzir os danos causados à mucosa intestinal, histoarquitetura dos rins e do fígado.

MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model

Abstract Purpose To investigate the role and related mechanisms of miR-106a in sepsis-induced AKI. Methods Serum from sepsis and healthy patients was collected, sepsis mouse model was established by cecal ligation and puncture (CLP). TCMK-1 cells were treated with lipopolysaccharide (LPS) and transfected with THBS2-small interfering RNA (siTHBS2), miR-106a inhibitor, miR-106a mimics and their negative controls (NCs). The expression of miR-106a, thrombospondin 2 (THBS2), Bax, cleaved caspase-3 and Bcl-2, cell viability, relative caspase-3 activity and TNF-α, IL-1β, IL-6 content were respectively detected by quantitative real-time polymerase chain reaction (qRT-PCR), western blotting, Cell Counting Kit-8 (CCK-8) and enzyme linked immunosorbent assay (ELISA). The relationship between miR-106a and THBS2 was confirmed by dual luciferase reporter assay. Results MiR-106a was up-regulated in serum of sepsis patients, CLP-induced mice models and LPS-induced TCMK-1 cells. LPS reduced cell viability and Bcl-2 expression, and increased caspase-3 activity, Bax expression, the content of TNF-α, IL-1β, IL-6. THBS2 was a target of miR-106a. The decreases of caspase-3 activity, TNF-α, IL-1β, IL-6, Bax expression and the increases of cell viability, Bcl-2 expression caused by miR-106a knockdown were reversed when THBS2 silencing in LPS-stimulated TCMK-1 cells. Conclusion MiR-106a aggravated LPS-induced inflammation and apoptosis of TCMK-1 cells via regulating THBS2 expression.

CXCL8(3-72) K11R/G31P protects against sepsis-induced acute kidney injury via NF-κB and JAK2/STAT3 pathway

BACKGROUND: Acute kidney injury (AKI), which is mainly caused by sepsis, has high morbidity and mortality rates. CXCL8(3-72) K11R/G31P (G31P) can exert therapeutic effect on inflammatory diseases and malignancies. We aimed to investigate the effect and mechanism of G31P on septic AKI. METHODS: An AKI mouse model was established, and kidney injury was assessed by histological analysis. The contents of serum creatinine (SCr) and blood urea nitrogen (BUN) were measured by commercial kits, whereas neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) were detected by enzyme-linked immunosorbent assay (ELISA) kits. The expressions of CXCL8 in serum and kidney tissues were determined using ELISA and immunohistochemical analysis, respectively. Apoptosis rate of renal tissue was detected by terminal deoxynucleotidyl transfer-mediated dUTP nick end labeling (TUNEL) analysis. The expressions of inflammatory cytokines were measured by quantitative real-time PCR and Western blot, respectively. The apoptosis-related proteins, JAK2, STAT3, NF-κB and IκB were determined by Western blot. RESULTS: G31P could reduce the levels of SCr, BUN, HGAL and KIM-1 and inhibit the renal tissue injury in AKI mice. G31P was also found to suppress the serum and nephric CXCL8 expressions and attenuated the apoptosis rate. The levels of inflammatory cytokines, pro-apoptotic proteins were decreased, while the anti-apoptotic proteins were increased by G31P in AKI mice. G31P also inhibited the activation of JAK2, STAT3 and NF-κB in AKI mice. CONCLUSION: These results suggest that G31P could protect renal function and attenuate the septic AKI. Our findings provide a potential target for the treatment of AKI.

Adiponectin is protective against endoplasmic reticulum stress-induced apoptosis of endothelial cells in sepsis

Endoplasmic reticulum (ER) stress is a critical molecular mechanism involved in the pathogenesis of sepsis. Hence, strategies for alleviating this stress may be essential for preventing cardiovascular injuries under sepsis. Adiponectin is secreted by adipocytes and its levels are decreased in sepsis. The purpose of this study was to investigate the protective effects of adiponectin treatment on endothelial cells and its mechanism. Male Wistar rats underwent cecal ligation and puncture (CLP) before being treated with adiponectin (72 and 120 μg/kg). The levels of malondialdehyde (MDA) in plasma, histological structure, and apoptosis of endothelial cells were evaluated. In vitro, human umbilical vein endothelial cells (HUVECs) were treated with adiponectin at 10 and 20 μg/mL for 24 h after stimulation by lipopolysaccharide (LPS). The levels of reactive oxygen species (ROS), ultrastructure, rate of apoptosis, the expression of inositol-requiring enzyme 1α (IRE1α) protein, and its downstream molecules (78 kDa glucose-regulated protein (GRP78), C/EBP homologous protein (CHOP), and caspase-12) were detected. The results showed that the levels of MDA and ROS induced by CLP or LPS stimulation were increased. Furthermore, endothelial cell apoptosis was increased under sepsis. The IRE1α pathway was initiated, as evidenced by activated IRE1α, increased GRP78, and up-regulated CHOP and caspase-12 in HUVECs. Following treatment with adiponectin, the number of apoptotic endothelial cells was markedly decreased. These findings demonstrated that treatment with adiponectin decreased apoptosis of endothelial cells caused by sepsis by attenuating the ER stress IRE1α pathway activated by oxidative stress.

The effect of tadalafil therapy on kidney damage caused by sepsis in a polymicrobial septic model induced in rats: a biochemical and histopathological study

ABSTRACT Introduction Sepsis is an inflammatory reaction to bacteria involving the whole body and is a significant cause of mortality and economic costs. The purpose of this research was to determine whether tadalafil exhibits a preventive effect on sepsis in a septic model induced in rats with cecal ligation and puncture (CLP). Materials and Methods Rats were randomly separated into groups, 10 rats in each: (i) a sham (control) group, (ii) an untreated sepsis group, (iii) a sepsis group treated with 5mg/kg tadalafil and (iv) a sepsis group treated with 10mg/kg tadalafil. A polymicrobial sepsis model was induced in rats using CLP. Rats were sacrificed after 16h, and blood and kidney tissues were collected for biochemical and histopathological study. Results Levels of the inflammatory parameter IL-6 decreased significantly in the sepsis groups receiving tadalafil in comparison with the untreated sepsis group (p<0.05). In terms of histopathology, inflammation scores investigated in kidney tissues decreased significantly in the sepsis groups receiving tadalafil compared to the untreated sepsis group (p<0.05). In addition, levels of creatinine and cystatin C measured in septic rats receiving tadalafil were lower by a clear degree than in septic rats (p<0.05). Conclusion In this study, tadalafil exhibited a preventive effect for sepsis-related damage by suppressing inflammation in serum and kidney tissue of septic rats in a polymicrobial sepsis model induced with CLP.

Anatomo-radiological correlation using 18-FDG-PET in abdominal sepsis model in rats. A preliminary study

Abstract Purpose: To examine a correlation of micro-PET images with photographic images of the digestive organs in abdominal sepsis model. Methods: Male Wistar rats weighing 265±18g were used. Abdominal sepsis was induced by ligature and cecal puncture. Micro-PET Images from abdominal cavity septic foci were obtained using 18-Fluoro-deoxyglucose, looking for a correlation with photographic images of abdominal cavity organs. Pearson's correlation test was used. Results: The mean standard uptake values (SUV) and lesion areas were 2.58±0.63SUVbwg/ml and 546.87±300.95mm2, respectively. There was a strong positive correlation between the two variables (r=0.863, p=0.137), which resulted in a coefficient of determination r2?0.75, meaning that 75% of SUV variation is explained by the lesion areas of digestive organs. Conclusion: Micro-PET allows high throughput assessment of lesion count and volume in pre-clinical rat model of CPL abdominal sepsis.

Déficit de base y depuración del lactato en el paciente pediátrico con sepsis / Base deficit and lactate depuration in pediatric septic patient

El shock séptico genera un estado de falla de aporte de oxígeno y nutrientes a la célula, consiguientemente liberación de productos como el lactato y otros ácidos orgánicos; produciendo un déficit de bases. OBJETIVO: el presente estudio pretende estimar la correlación del lactato sérico y el déficit de bases como guías en el manejo del shock séptico durante las primeras 24 horas METODOS:se realizó un estudio observacional prospectivo; realizando la medición seriada de lactato y del déficit de base; en pacientes shock séptico tratados de acuerdo a recomendaciones de la Campaña Sobreviviendo a la Sepsis (CSS) 2013. RESULTADOS: el lactato sérico y el déficit de base se depuran en 40% y 5% respectivamente durante las primeras 24 horas; no existe correlación (p=0.289) al ingreso, observándose posteriormente una correlación significativa (p=0,000). Los niveles de depuración en los pacientes fallecidos fueron menores en relación a los sobrevivientes. CCONCLUSIONES: el déficit de base no debe ser considerado como un sustituto fiable para el lactato sérico en el paciente con shock séptico; aunque probablemente sea de utilidad después de las 24 horas.

Sepsis and septic shock generate a state of microcirculatory failure, hipoxia and anaerobic metabolism and products like lactic acid and other organic acids are released; which lead to a deficit of bases. OBJECTIVE: this study aims to estimate the correlation of serum lactate and base deficit as guides in the management of septic shock during the first 24 hours. METHOD: a prospective observational study; performing serial measurement hours 0, 6, 12 and 24 of lactate and base deficit; in patients with the same scheme of treatment for septic shock recommended by the CSS 2013. RESULTS: the serum lactate and base deficit were modified with resuscitation with a clearance of 40% and 5% respectively; we note that there is no correlation between values (p = 0.289) at Hr. 0; However at 24 hours a significant correlation (P = 0.000) is observed. CONCLUSIONS: the base deficit should not be considered a reliable substitute for serum lactate at the beginning of resuscitation of patients with sepsis, severe sepsis and septic shock lactate; but it is likely useful after 24 hours.

Determinantes precoces en el desarrollo de injuria renal aguda durante la sepsis abdominal experimental / Early determinants of acute kidney injury during experimental intra-abdominal sepsis

Background: Sepsis-induced acute kidney injury (AKI) is an early and frequent organ dysfunction, associated with increased mortality. Aim: To evaluate the impact of macrohemodynamic and microcirculatory changes on renal function and histology during an experimental model of intra-abdominal sepsis. Material and Methods: In 18 anaesthetized pigs, catheters were installed to measure hemodynamic parameters in the carotid, right renal and pulmonary arteries. After baseline assessment and stabilization, animals were randomly divided to receive and intra-abdominal infusion of autologous feces or saline. Animals were observed for 18 hours thereafter. Results: In all septic animals, serum lactate levels increased, but only eight developed AKI (66%). These animals had higher creatinine and interleukin-6 levels, lower inulin and para-aminohippurate clearance (decreased glomerular filtration and renal plasma flow), and a negative lactate uptake. Septic animals with AKI had lower values of mean end arterial pressure, renal blood flow and kidney perfusion pressure, with an associated increase in kidney oxygen extraction. No tubular necrosis was observed in kidney histology. Conclusions: The reduction in renal blood flow and renal perfusion pressure were the main mechanisms associated with AKI, but were not associated with necrosis. Probably other mechanisms, such as microcirculatory vasoconstriction and inflammation also contributes to AKI development.

Median lethal needle caliber in two models of experimental sepsis

To estimate the median lethal needle caliber (LC50) of a new experimental sepsis model and compare it to the LC50 of the cecal ligation and puncture (CLP) sepsis model. METHODS: Male albino Wistar rats were studied (n=22). Animals were allocated into two study groups. In Group I, experimental sepsis was induced by cecal ligation and puncture. In Group II, experimental sepsis was induced by ascending colon ligation and cecal puncture. Up-and-down method was used to determinate the LC50. RESULTS: LC50 in Group I was 19 Gauge (Confidence Interval 17 to 22 Gauge). Determination of LC50 was not possible in Group II due to the death of all animals. CONCLUSION: LC50 in cecal ligation and puncture is 19 Gauge. The lethality of the new model tested in this trial is very high.

Validation of a Modified Early Warning Score to Predict ICU Transfer for Patients with Severe Sepsis or Septic Shock on General Wards

PURPOSE: To assess whether the Modified Early Warning Score (MEWS) predicts the need for intensive care unit (ICU) transfer for patients with severe sepsis or septic shock admitted to general wards. METHODS: A retrospective chart review of 100 general ward patients with severe sepsis or septic shock was implemented. Clinical information and MEWS according to point of time between ICU group and general ward group were reviewed. Data were analyzed using multivariate logistic regression and the area under the receiver operating characteristic curves with SPSS/WIN 18.0 program. RESULTS: Thirty-eight ICU patients and sixty-two general ward patients were included. In multivariate logistic regression, MEWS (odds ratio [OR] 2.02, 95% confidence interval [CI] 1.43-2.85), lactic acid (OR 1.83, 95% CI 1.22-2.73) and diastolic blood pressure (OR 0.89, 95% CI 0.80-1.00) were predictive of ICU transfer. The sensitivity and the specificity of MEWS used with cut-off value of six were 89.5% and 67.7% for ICU transfer. CONCLUSION: MEWS is an effective predictor of ICU transfer. A clinical algorithm could be created to respond to high MEWS and intervene with appropriate changes in clinical management.

Septicemia decorrente de pododermatites em canários de cor (Serinus canarius) / Septicemia resulting from bumblefoot in color canaries (Serinus canarius)

Relataram-se um surto de pododermatite e quadro septicêmico em aves de um canaril comercial. Quarenta e quatro canários de cor foram escolhidos de forma aleatória, sem distinção de sexo, idade ou cor, os quais vieram a óbito naturalmente, após terem sido afetados pela pododermatite, sem que tivessem se submetido a tratamento prévio. As aves mortas foram encaminhadas para o exame necroscópico, onde amostras de tecidos das áreas afetadas foram colhidas para exames microbiológico, micológico e histológico. Todas as aves necropsiadas apresentavam pododermatite, com inflamação em um ou mais dedos, de aspecto nodular, com ou sem presença de úlceras ou necrose. Staphylococcus aureus plasma coagulase positivo foi isolado dos pés e do fígado de todas as aves. Foi observado que todas as linhagens isoladas foram resistentes aos antimicrobianos da classe das penicilinas (penicilina G e ampicilina) e parcialmente sensíveis ou resistentes à ciprofloxacina. Apenas metade dos isolados foram sensíveis à neomicina e à estreptomicina. Problemas de saúde pública podem estar relacionados ao surgimento de animais reservatórios de cepas multirresistentes para seres humanos contactantes, como neste caso.

An outbreak of bumblefoot and septicemia was reported in birds of commercial breeding. Forty-four color canaries chosen at random, without regard to sex, age or color, which died naturally having been affected with pododermatitis and not undergoing any prior treatment were used. The dead birds were sent for necropsy in which tissue samples from affected areas were taken for microbiological, mycological and histological examination. All necropsied birds had bumblefoot, inflammation in one or more fingers, nodular, with or without the presence of ulcers and necrosis. Staphylococcus aureus plasma-coagulase positive was isolated from the liver and the feet of all birds. It was observed that all isolates were resistant to the penicillin class of antibiotics (penicillin G and ampicillin) and partially sensitive or resistant to ciprofloxacin. Only half of the isolates were sensitive to neomycin and streptomycin. Public health problems may be related to the emergence of animal reservoirs of multi-resistant strains for contacted humans, as in this case.

Study of bacteria isolated from the foot pad of Spheniscus magellanicus with and without bumblefoot / Estudo de bactérias isoladas de coxins plantares de Spheniscus magellanicus com e sem bumblefoot

The bumblefoot or pododermatitis is among the diseases with the highest morbidity in Magellanic penguins, sometimes evolving to septicemia and death. Therefore, this study aimed to relate the main species involved in the disorder, as well as the in vitro susceptibility profile of the microorganisms against routine antimicrobial usage in Veterinary Medicine. During two years in vivo material was harvested from 200 footpads (n=100 animals) for microbiological analysis and in vitro susceptibility tests against the Antibiotic enrofloxacin, streptomycin, penicillin and cephalosporin. Bacteria have been identified both as part of permanent and transient microbiota, also being associated to 100% of the pododermatitis cases. The most prevalent genus were Staphylococcus and Corynebacterium. The antibiograms of all the isolated bacteria resulted in greater susceptibility of the strains facing cephalosporin, followed by enrofloxacin, streptomycin and penicillin.

O bumblefoot ou pododermatite está entre as afecções de maior morbidade em pinguins-de-magalhães, podendo evoluir para septicemia e óbito. Portanto, o presente estudo objetivou relacionar as principais espécies bacterianas envolvidas na afecção, bem como o perfil de susceptibilidade in vitro destes microrganismos frente a antimicrobianos de uso rotineiro em medicina veterinária. Durante o período de dois anos, foi realizada colheita de material in vivo de 200 coxins plantares (n=100 animais) para análise microbiológica e testes de susceptibilidade in vitro frente aos antibióticos enrofloxacina, estreptomicina, penicilina e cefalosporina. Bactérias foram identificadas tanto como parte da microbiota permanente quanto da transitória, bem como estiveram associadas a 100% dos casos de pododermatite. Os gêneros mais prevalentes foram Staphylococcus e Corynebacterium. Os antibiogramas de todas as bactérias isoladas resultaram em maior sensibilidade das cepas frente à cefalosporina, seguida de enrofloxacina, estreptomicina e penicilina.

Edwardsiella tarda sepsis with multiple liver abscesses in a patient with Cushing's syndrome.

Edwardsiella tarda is very seldom a cause for gastroenteritis in humans. This organism can also cause extraintestinal infections, such as soft tissue infections, meningitis, peritonitis, osteomyelitis, endocarditis and hepatobiliary tract disease, particularly in the setting of compromised immunity. We describe, for the first time a case of E. tarda sepsis with multiple liver abscesses associated with Cushing's syndrome as a result of recreational aquatic exposure.

Non invasive real-time monitoring of bacterial infection & therapeutic effect of anti-microbials in five mouse models.

Background & objectives: In vivo imaging system has contributed significantly to the understanding of bacterial infection and efficacy of drugs in animal model. We report five rapid, reproducible, and non invasive murine pulmonary infection, skin and soft tissue infection, sepsis, and meningitis models using Xenogen bioluminescent strains and specialized in vivo imaging system (IVIS). Methods: The progression of bacterial infection in different target organs was evaluated by the photon intensity and target organ bacterial counts. Genetically engineered bioluminescent bacterial strains viz. Staphylococcus aureus Xen 8.1, 29 and 31; Streptococcus pneumoniae Xen 9 and 10 and Pseudomonas aeruginosa Xen-5 were used to induce different target organs infection and were validated with commercially available antibiotics. Results: The lower limit of detection of colony forming unit (cfu) was 1.7-log10 whereas the lower limit of detection of relative light unit (RLU) was 4.2-log10. Recovery of live bacteria from different target organs showed that the bioluminescent signal correlated to the live bacterial count. Interpretation & conclusions: This study demonstrated the real time monitoring and non-invasive analysis of progression of infection and pharmacological efficacy of drugs. These models may be useful for pre-clinical discovery of new antibiotics.

Sepse: diagnóstico e tratamento / Sepsis: diagnosis and treatment

JUSTIFICATIVA E OBJETIVOS: A sepse é um importante problema de saúde no Brasil, incluindo serviços públicos e privados, com altos custos de tratamento e mortalidade. Desde 2003, a Campanha Sobrevivendo a Sepse vem sugerindo a implementação de medidas para o tratamento desta condição mórbida, de modo a reduzir sua mortalidade em 25%. O objetivo deste estudo foi contextualizar a atividade do emergencista no tratamento da sepse, bem como revisar os principais aspectos da epidemiologia da sepse no Brasil e seus tratamentos com base na Campanha Sobrevivendo a Sepse. CONTEÚDO: Utilizando as palavras-chave sepse, Surviving Sepsis Campaign, epidemiologia, fisiopatologia foi procedida a busca de referências no PubMed, Scopus, SciElo. Na Campanha Sobrevivendo à Sepse, foi utilizado um sistema de notas para orientar a avaliação de qualidade e evidência de alta (A) a muito baixa (D) e, deste modo, determinar a força de recomendações. Neste artigo foram registradas estas recomendações como: recomendação forte [1] indica os efeitos desejáveis da intervenção claramente superiores aos seus efeitos indesejáveis, ou claramente não; recomendação fraca [2] indica que a distinção entre os efeitos desejáveis e indesejáveis é menos clara. CONCLUSÃO: O clinico e, sobretudo o clínico emergencista, exerce papel central no tratamento da sepse na medida em que novas evidencias demonstram que a precocidade do tratamento é peça-chave no sucesso do tratamento.

BACKGROUND AND OBJECTIVES: Sepsis remains an important international health problem including Brazil, with high treatment costs and unacceptable morbidity and mortality. Since 2003, the Sepsis Surviving Campaign was created and suggested guidelines for sepsis and the campaign objective is reducing sepsis mortality rate around 25%. In this aim this work reviews the most important aspects of sepsis diagnosis and treatment, especially in the emergencist physician context and abording Brazilian epidemiology aspects. CONTENTS: This overview was performed using sepsis, septic shock, Surviving Sepsis Campaign, sepsis epidemiology and fisiopathology as keywords in PubMed, Scopus and Scielo databases. In Surviving Sepsis Campaign, was used a grade system to guide assessment to quality and evidence from high (A) to very low (D) and, at this way, determine the strength of recommendations. In this article this recommendations was registered as a follow: strong recommendation [1] indicates the intervention?s desirable effects clearly outweigh its undesirable effects, or clearly do not; weak recommendations [2] indicate that the tradeoff between desirable and undesirable effects is less clear. CONCLUSION: Physicians, even emergence physicians, plays a central role in the modern management of sepsis and septic shock, across the golden hours of treatment, which ones new evidences demonstrate the importance of nearly approach of patient's results in sepsis mortality decrease