Peculiarities of the phenotype of T-lymphocytes in the dynamics of the postoperating period in patients with peritonite depending on the outcome of disease

The aim of the study was to investigate the phenotype of blood T-lymphocytes in the dynamics of the postoperative period in patients with widespread purulent peritonitis (WPP) depending on the outcome of the disease. A total of 36 patients aged 30–65 with acute surgical diseases and abdominal injuries complicated by WPP years were examined. Blood sampling was performed before the operation (preoperative period), and also on the 7th, 14th and 21st day of the postoperative period. 40 relatively healthy people of the same age range were examined as a control. Study of the phenotype of blood T-lymphocytes was carried out by the method of 5-color f low cytometry using direct immunof luorescence of whole peripheral blood. Mean levels of fluorescence were used to evaluate the expression levels of surface receptors. It was established that in the preoperative period in patients with WPP regardless of the outcome of the disease against the background of a decrease in the absolute amount of total lymphocytes in the blood the content of T cells increases. The T-lymphocytes of patients with WPP are significantly more intense expressing the CD28 and CD62L receptors than the cells of healthy people. In the preoperative period and within two weeks of postoperative treatment with a favorable outcome of the WPP an increased amount of CD62L+T-lymphocytes is detected in comparison with the indices of patients with an unfavorable outcome of the disease. Other features of the phenotypic composition of T-lymphocytes in patients with a favorable outcome of WPP is an increase in T-regulatory activity which manifests itself both in the preoperative period and within two weeks of postoperative treatment. With a favorable outcome of WPP an increase in the number of activated cytotoxic T-lymphocytes is observed on the 14th day of treatment which with an unfavorable outcome is observed only on the 21st day of treatment. It is assumed that T-cell suppression and activation of cytotoxic T cells are the factors determining a decrease in the intensity of inflammatory processes in WPP and thereby increasing the level of the favorable outcome of the disease. In case of an unfavorable outcome of the disease, postoperative therapy has a weaker or delayed effect on the dynamics of the studied parameters than with a favorable outcome. With a favorable outcome of WPP an increase in the CD3+CD57+cell count is observed already from the second week of treatment and is more pronounced whereas in the case of an adverse outcome of WPP an increase in the level of these cells is observed only at the third week of treatment. However, the level of expression of CD57 on T-lymphocytes is more pronounced throughout the course of the examination with an unfavorable outcome of WPP

REGULATORY INFLUENCE OF BLOOD MONOCYTES ON THE POPULATION COMPOSITION OF GRANULOCYTES AND THE STATE OF THEIR RESPIRATORY BURST IN THE WIDESPREAD PURULENT PERITONITIS

The aim of the study was to investigate the regulatory effect of monocytes and their subpopulations on the population composition  of granulocyte leukocytes and the state of their respiratory burst in  widespread purulent peritonitis (WPP). The study involved 24  patients aged 30-65 with acute surgical diseases and injuries of  abdominal organs complicated by WPP. As a control 25 relatively  healthy people of the same age range were examined. A study of the population composition of monocytes and granulocyte leukocytes in  blood was performed using a two-platform technology on the  hematological analyzer Sysmex XE-5000 (Sysmex Inc., USA) and  FC-500 flow cytometer (Beckman Coulter, USA) using the Cytodiff antibody kit (Beckman Coulter, USA). A study of the  monocytes number expressing HLA-DR- and CD64-receptor was performed by flow cytometry using direct immunofluorescence  of whole peripheral blood. The respiratory burst state of neutrophilic granulocytes was studied by chemiluminescence analysis on a 36-channel chemiluminescence analyzer BLM-3607 (MedBioTech, Russia). As indicators of chemiluminescence  were used luminol and lucigenin. The enhancement of  chemiluminescence induced by zymosan was evaluated by the ratio  of the area of the induced chemiluminescence to the spontaneous area and was defined as the activation index. It has  been established that the immune-inflammatory process in WPP is  characterized by a decrease in the number of classical monocytes in  the peripheral blood and an increase in the content of non-classical  monocytes. In WPP in peripheral blood the level of monocytes  expressing HLA-DR receptors decreases. The change in the ratio of  monocytes subpopulations characterizes the increase in the role of  the proinflammatory fraction in the WPP pathogenesis. Changes in  the population composition of granulocytes in the blood in patients  with WPP also characterize the development of an acute  inflammatory process. In this case, there is a decrease in the  number of basophils in the peripheral blood, which, apparently, is  determined by the presence of an allergic component in WPP and,  accordingly, their migration to the inflammation area. In patients  with WPP activation of a respiratory burst of granulocytes of blood  was detected, the intensity of which is determined by the synthesis  of primary and secondary active oxygen species. The results of the  correlation analysis made it possible to establish that in WPP the  regulatory role of non-classical monocytes increases aimed at  stimulating the inflammatory processes (an increase in the number  of mature and immature forms of neutrophils and stimulation of the activity of a respiratory explosion of granulocytes). The revealed features of the regulatory effect of monocytes on the population  composition and the intensity of the respiratory burst of granulocytes can be used in the development of immunotherapeutic methods aimed at reducing the activity of the inflammatory process in WPP

FEATURES OF THE PHENOTYPE AND NAD(P)-DEPENDENT DEHYDROGENASES ACTIVITY IN NEUTROPHIL BY PATIENTS WITH WIDESPREAD PURULENT PERITONITIS IN PROGNOSIS FOR SEPSIS DEVELOPMENT

The aim of the study was to examine the features of phenotype and the levels of NAD(P)-dependent dehydrogenases activity of blood neutrophils in the prognosis of abdominal sepsis development in patients with widespread purulent peritonitis (WPP). 50 patients with WPP of community and hospital origin in the pre-operative period were examined. From the 5th to the 10th day of the postoperative period 35 patients (70%) had developed abdominal sepsis, 15 patients (30%) had absence of complications. As controls 67 respect healthy people were examined. The research blood neutrophils phenotype was performed by f low cytometry using a direct immunofluorescence whole peripheral blood. The levels of surface receptor expression was assessed by the mean fluorescence intensity. The NAD(P)-dependent dehydrogenases activity in the blood neutrophils studied using bioluminescence method. It was established that the inflammatory reaction in patients with WPP is characterized by neutrophilia and changes in the phenotype of blood neutrophils. The markers of the subsequent development of sepsis in WPP are less pronounced (in comparison with the indices of uncomplicated patients), an increase in the number of neutrophils, a decrease in the HLA-DR + -cell count against the background of a high level of neutrophils expressing a high affinity receptor for IgG. The patients without subsequent complications had the number of neutrophils with CD23 receptor expression is increased. Metabolism of neutrophils in patients with WPP is characterized by a decrease in the intensity of plastic processes due to low activity of glucose-6-phosphate dehydrogenase and an imbalance in the activity of the enzymes of the mitochondrial compartment. A feature of the neutrophil metabolism in patients with WPP without subsequent development of sepsis is a high activity of anaerobic lactate dehydrogenase reaction and a decrease in the activity of NADP-dependent decarboxylating malate dehydrogenase. The patients with WPP with subsequent development of sepsis had a high level of NAD-dependent out flow of substrates from the tricarboxylic acid cycle on the amino acid exchange reaction via glutamate dehydrogenase which can affect the activity of aerobic respiration of blood neutrophils. The established differences in the phenotype and activity of enzymes in the blood neutrophils in patients with WPP depending on the subsequent development of sepsis determine the possibility of creating a method for predicting the development of complications and developing immunoactive therapy in the postoperative period of WPP

THE PHENOTYPE AND METABOLISM RELATIONSHIP OF BLOOD NEUTROPHILS IN PATIENTS WITH WIDESPREAD PURULENT PERITONITIS IN THE POSTOPERATIVE PERIOD DYNAMICS

The aim of the study was to examine the relationship of the phenotype and metabolism of neutrophils in patients with widespread purulent peritonitis (WPP) in the dynamics of the postoperative period. The study involved 27 patients with acute surgical diseases and injuries of abdominal organs complicated by WPP. Blood sampling was performed prior to surgery (pre-operative period) and at 7, 14 and 24 day post-operative period. As controls 67 respect healthy people were examined. Research blood neutrophilic granulocytes phenotype was performed by f low cytometry using a direct immunof luorescence whole peripheral blood. The levels of surface receptor expression was assessed by the mean f luorescence intensity. The NADand NADP-dependent dehydrogenases activity in the blood neutrophils studied using bioluminescence method. It was found that in patients with WPP in the preoperative period in the peripheral blood increased content of CD62L+-, HLA-DR+and CD64+-neutrophils. High levels of CD62L+-cells stored within 24 postoperative days, whereas the amount of HLA-DR+and CD64+-neutrophils on 24 postoperative day is reduced to the level of controls. The dynamics of changes in the content of CD64+-cells in the peripheral blood of patients with WPP corresponds to the expression level of CD64-receptor on the membrane of neutrophilic granulocytes. The metabolism of blood neutrophils in patients with WPP in the preand postoperative period is characterized by high intensity of the substrate stream on the citric acid cycle, low activity of NADP-dependent glutamate dehydrogenase and aerobic reaction of lactate dehydrogenase. In the preoperative period and within 14 days of the postoperative period in neutrophil granulocytes of the patients revealed high activity of anaerobic lactate dehydrogenase reaction characterized by increased activity of anaerobic glycolysis. In the late postoperative period the intensity of anaerobic energy in the neutrophils of patients with WPP was reduced to the control level. The final stage of post-operative treatment the metabolism in the blood neutrophils of patients with WPP is also characterized by the activation of the pentose phosphate cycle, low activity of malate-aspartate shunt mitochondria and high intensity of the substrate interaction between citric acid cycle and reactions of amino acid metabolism. Using correlation analysis set dysregulation between phenotype and the system of intracellular metabolism of neutrophils which can be determined by the migration of activated cells in the inf lammatory focus as well as changes in the activity of intracellular enzymes under different regulatory factors and including postoperative therapy methods peritonitis

PHAGOCYTIC ACTIVITY AND BLOOD NEUTROPHILS RESPIRATORY BURST STATE FEATURES AMONGST WIDESPREAD PURULENT PERITONITIS PATIENTS IN THE POSTOPERATIVE PERIOD DYNAMICS

The research aim was to study the phagocytic activity and the blood neutrophils respiratory burst state amongst the patients with widespread purulent peritonitis (WPP) in the dynamics of the postoperative period. The study was involved patients with acute surgical diseases and injuries of the abdominal organs complicated by WPP. Blood sampling was performed prior to surgery (pre-operative period) and at 7, 14 and 24 day post-operative period. The neutrophil phagocytosis level was determined by flow cytometry using FITC-labeled staphylococcal protein A. There was counted the percentage of fluorescent neutrophils (defined as the phagocytic index — PI) and average of the cell fluorescence (phagocytic number — PN). According to the fluorescence intensity divided phagocytic neutrophils in an active (high PN) and weakly phagocytic (low PN). Condition of the respiratory burst in the neutrophils was examined using a chemiluminescent assay. It was found that patients with WPP in the preoperative period had increased phagocytic activity of the blood neutrophils. The maximum phagocytic activity observed on day 7 of the postoperative period and by 24 days the number of phagocytic cells is reduced to control level whereas phagocytic activity of the cells remains at the preoperative level. In the preoperative period in patients with WPP the amount of active phagocytic blood neutrophils 3.3 times higher than the number of weakly phagocytic cells. By the end of the observation period the number of active and weakly phagocytic neutrophils returning to normal. In the study of the state of respiratory burst in the neutrophils by the WPP patients discovered that the primary synthesis levels of reactive oxygen species (ROS) increases only with 7 days of postoperative period and the induced activity of NADPH-oxidase remains elevated to the end of the observation period. At the same time, the synthesis of secondary ROS in the blood neutrophil of the WPP patients increased already in the preoperative period and remains high until the end of the survey. Using correlation analysis it was found that if healthy people had a competitive relationship between the level of synthesis of primary ROS and the activity of phagocytosis then all detected correlations in the WPP patients characterized by high level of coordination between the respiratory burst and the phagocytic activity of the neutrophils. These relationships are identified in the preoperative period, postoperatively do not exist. It can be assumed that the surgery and the subsequent treatment leads to complete incoordination in the neutrophils the mechanisms of phagocytosis and respiratory burst

An international assessment of the adoption of enhanced recovery after surgery (ERAS®) principles across colorectal units in 2019–2020

Aim: The Enhanced Recovery After Surgery (ERAS®) Society guidelines aim to standardize perioperative care in colorectal surgery via 25 principles. We aimed to assess the variation in uptake of these principles across an international network of colorectal units. Method: An online survey was circulated amongst European Society of Coloproctology members in 2019–2020. For each ERAS principle, respondents were asked to score how frequently the principle was implemented in their hospital, from 1 (‘rarely’) to 4 (‘always’). Respondents were also asked to recall whether practice had changed since 2017. Subgroup analyses based on hospital characteristics were conducted. Results: Of hospitals approached, 58% responded to the survey (195/335), with 296 individual responses (multiple responses were received from some hospitals). The majority were European (163/195, 83.6%). Overall, respondents indicated they ‘most often’ or ‘always’ adhered to most individual ERAS principles (18/25, 72%). Variability in the uptake of principles was reported, with universal uptake of some principles (e.g., prophylactic antibiotics; early mobilization) and inconsistency from ‘rarely’ to ‘always’ in others (e.g., no nasogastric intubation; no preoperative fasting and carbohydrate drinks). In alignment with 2018 ERAS guideline updates, adherence to principles for prehabilitation, managing anaemia and postoperative nutrition appears to have increased since 2017. Conclusions: Uptake of ERAS principles varied across hospitals, and not all 25 principles were equally adhered to. Whilst some principles exhibited a high level of acceptance, others had a wide variability in uptake indicative of controversy or barriers to uptake. Further research into specific principles is required to improve ERAS implementation

An international assessment of the adoption of enhanced recovery after surgery (ERAS®) principles across colorectal units in 2019–2020

AimThe Enhanced Recovery After Surgery (ERAS®) Society guidelines aim to standardize perioperative care in colorectal surgery via 25 principles. We aimed to assess the variation in uptake of these principles across an international network of colorectal units.MethodAn online survey was circulated amongst European Society of Coloproctology members in 2019–2020. For each ERAS principle, respondents were asked to score how frequently the principle was implemented in their hospital, from 1 (‘rarely’) to 4 (‘always’). Respondents were also asked to recall whether practice had changed since 2017. Subgroup analyses based on hospital characteristics were conducted.ResultsOf hospitals approached, 58% responded to the survey (195/335), with 296 individual responses (multiple responses were received from some hospitals). The majority were European (163/195, 83.6%). Overall, respondents indicated they ‘most often’ or ‘always’ adhered to most individual ERAS principles (18/25, 72%). Variability in the uptake of principles was reported, with universal uptake of some principles (e.g., prophylactic antibiotics; early mobilization) and inconsistency from ‘rarely’ to ‘always’ in others (e.g., no nasogastric intubation; no preoperative fasting and carbohydrate drinks). In alignment with 2018 ERAS guideline updates, adherence to principles for prehabilitation, managing anaemia and postoperative nutrition appears to have increased since 2017.ConclusionsUptake of ERAS principles varied across hospitals, and not all 25 principles were equally adhered to. Whilst some principles exhibited a high level of acceptance, others had a wide variability in uptake indicative of controversy or barriers to uptake. Further research into specific principles is required to improve ERAS implementation.AimThe Enhanced Recovery After Surgery (ERAS®) Society guidelines aim to standardize perioperative care in colorectal surgery via 25 principles. We aimed to assess the variation in uptake of these principles across an international network of colorectal units.MethodAn online survey was circulated amongst European Society of Coloproctology members in 2019–2020. For each ERAS principle, respondents were asked to score how frequently the principle was implemented in their hospital, from 1 (‘rarely’) to 4 (‘always’). Respondents were also asked to recall whether practice had changed since 2017. Subgroup analyses based on hospital characteristics were conducted.ResultsOf hospitals approached, 58% responded to the survey (195/335), with 296 individual responses (multiple responses were received from some hospitals). The majority were European (163/195, 83.6%). Overall, respondents indicated they ‘most often’ or ‘always’ adhered to most individual ERAS principles (18/25, 72%). Variability in the uptake of principles was reported, with universal uptake of some principles (e.g., prophylactic antibiotics; early mobilization) and inconsistency from ‘rarely’ to ‘always’ in others (e.g., no nasogastric intubation; no preoperative fasting and carbohydrate drinks). In alignment with 2018 ERAS guideline updates, adherence to principles for prehabilitation, managing anaemia and postoperative nutrition appears to have increased since 2017.ConclusionsUptake of ERAS principles varied across hospitals, and not all 25 principles were equally adhered to. Whilst some principles exhibited a high level of acceptance, others had a wide variability in uptake indicative of controversy or barriers to uptake. Further research into specific principles is required to improve ERAS implementation.A

An international assessment of the adoption of enhanced recovery after surgery (ERAS®) principles across colorectal units in 2019–2020

Aim: The Enhanced Recovery After Surgery (ERAS®) Society guidelines aim to standardize perioperative care in colorectal surgery via 25 principles. We aimed to assess the variation in uptake of these principles across an international network of colorectal units. Method: An online survey was circulated amongst European Society of Coloproctology members in 2019–2020. For each ERAS principle, respondents were asked to score how frequently the principle was implemented in their hospital, from 1 (‘rarely’) to 4 (‘always’). Respondents were also asked to recall whether practice had changed since 2017. Subgroup analyses based on hospital characteristics were conducted. Results: Of hospitals approached, 58% responded to the survey (195/335), with 296 individual responses (multiple responses were received from some hospitals). The majority were European (163/195, 83.6%). Overall, respondents indicated they ‘most often’ or ‘always’ adhered to most individual ERAS principles (18/25, 72%). Variability in the uptake of principles was reported, with universal uptake of some principles (e.g., prophylactic antibiotics; early mobilization) and inconsistency from ‘rarely’ to ‘always’ in others (e.g., no nasogastric intubation; no preoperative fasting and carbohydrate drinks). In alignment with 2018 ERAS guideline updates, adherence to principles for prehabilitation, managing anaemia and postoperative nutrition appears to have increased since 2017. Conclusions: Uptake of ERAS principles varied across hospitals, and not all 25 principles were equally adhered to. Whilst some principles exhibited a high level of acceptance, others had a wide variability in uptake indicative of controversy or barriers to uptake. Further research into specific principles is required to improve ERAS implementation

Safety of primary anastomosis following emergency left sided colorectal resection: an international, multi-centre prospective audit.

This is the peer reviewed version of the following article: group, T. E. S. o. C. c. (2018). "Safety of primary anastomosis following emergency left sided colorectal resection: an international, multi-centre prospective audit." Colorectal Disease 20(S6): 47-57., which has been published in final form at https://doi.org/10.1111/codi.1437. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived VersionsINTRODUCTION: Some evidence suggests that primary anastomosis following left sided colorectal resection in the emergency setting may be safe in selected patients, and confer favourable outcomes to permanent enterostomy. The aim of this study was to compare the major postoperative complication rate in patients undergoing end stoma vs primary anastomosis following emergency left sided colorectal resection. METHODS: A pre-planned analysis of the European Society of Coloproctology 2017 audit. Adult patients (> 16 years) who underwent emergency (unplanned, within 24 h of hospital admission) left sided colonic or rectal resection were included. The primary endpoint was the 30-day major complication rate (Clavien-Dindo grade 3 to 5). RESULTS: From 591 patients, 455 (77%) received an end stoma, 103 a primary anastomosis (17%) and 33 primary anastomosis with defunctioning stoma (6%). In multivariable models, anastomosis was associated with a similar major complication rate to end stoma (adjusted odds ratio for end stoma 1.52, 95%CI 0.83-2.79, P = 0.173). Although a defunctioning stoma was not associated with reduced anastomotic leak (12% defunctioned [4/33] vs 13% not defunctioned [13/97], adjusted odds ratio 2.19, 95%CI 0.43-11.02, P = 0.343), it was associated with less severe complications (75% [3/4] with defunctioning stoma, 86.7% anastomosis only [13/15]), a lower mortality rate (0% [0/4] vs 20% [3/15]), and fewer reoperations (50% [2/4] vs 73% [11/15]) when a leak did occur. CONCLUSIONS: Primary anastomosis in selected patients appears safe after left sided emergency colorectal resection. A defunctioning stoma might mitigate against risk of subsequent complications

The impact of conversion on the risk of major complication following laparoscopic colonic surgery: an international, multicentre prospective audit.

This is the peer reviewed version of the following article: The and E. S. o. C. c. groups (2018). "The impact of conversion on the risk of major complication following laparoscopic colonic surgery: an international, multicentre prospective audit." Colorectal Disease 20(S6): 69-89., which has been published in final form at https://doi.org/10.1111/codi.14371. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.BACKGROUND: Laparoscopy has now been implemented as a standard of care for elective colonic resection around the world. During the adoption period, studies showed that conversion may be detrimental to patients, with poorer outcomes than both laparoscopic completed or planned open surgery. The primary aim of this study was to determine whether laparoscopic conversion was associated with a higher major complication rate than planned open surgery in contemporary, international practice. METHODS: Combined analysis of the European Society of Coloproctology 2017 and 2015 audits. Patients were included if they underwent elective resection of a colonic segment from the caecum to the rectosigmoid junction with primary anastomosis. The primary outcome measure was the 30-day major complication rate, defined as Clavien-Dindo grade III-V. RESULTS: Of 3980 patients, 64% (2561/3980) underwent laparoscopic surgery and a laparoscopic conversion rate of 14% (359/2561). The major complication rate was highest after open surgery (laparoscopic 7.4%, converted 9.7%, open 11.6%, P < 0.001). After case mix adjustment in a multilevel model, only planned open (and not laparoscopic converted) surgery was associated with increased major complications in comparison to laparoscopic surgery (OR 1.64, 1.27-2.11, P < 0.001). CONCLUSIONS: Appropriate laparoscopic conversion should not be considered a treatment failure in modern practice. Conversion does not appear to place patients at increased risk of complications vs planned open surgery, supporting broadening of selection criteria for attempted laparoscopy in elective colonic resection