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1.
Am Surg ; 89(12): 6134-6146, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37295804

ABSTRACT

BACKGROUND/OBJECTIVE (S): Completion pancreatectomy (C.P.) is one acceptable treatment of choice in clinical scenarios such as management of post-pancreatectomy complications and recurrence in the pancreatic remnant. Studies referring to completion pancreatectomy as a distinct operation are limited, without emphasizing at the operation itself, rather reporting completion pancreatectomy as a possible option for treatment of various diseases. The identification of indications of CP in various pathologies and the clinical outcomes are therefore mandatory. METHODS: A systematic literature search was performed in the Pubmed and Scopus Databases (February 2020),guided by the PRISMA protocol, for all studies reporting CP as a surgical procedure with reference at indications for performing it combined with postoperative morbidity and/or mortality. RESULTS: Out of 1647 studies, 32 studies from 10 countries with 2775 patients in total, of whom 561 (20.2%) CPs met the inclusion criteria and were included in the analysis. Inclusion year ranged from 1964 to 2018 and were published from 1992 until 2019. 17 studies with a total number of 249 CPs were performed for post-pancreatectomy complications. Mortality rate was 44.5% (111 out of 249). Morbidity rate was (72.6%). 12 studies with 225 CPs were performed for isolated local recurrence after initial resection with a morbidity rate of 21.5% and 0% mortality rate in the early postoperative period. Two studies with a total number of 12 patients reported CP as a treatment option for recurrent neuroendocrine neoplasms. The mortality in those studies was 8% (1/12) and the mean morbidity rate was 58.3% (7/12). Finally, CP for refractory chronic pancreatitis was presented in one study with morbidity and mortality rates of 19% and 0%, respectively. CONCLUSION: Completion pancreatectomy is a distinct treatment option for various pathologies. Morbidity and mortality rates depend on the indications of performing CP, the status performance of the patients and whether the operation is performed electively or urgently.


Subject(s)
Pancreatic Neoplasms , Pancreatitis, Chronic , Humans , Pancreatectomy/methods , Pancreatic Neoplasms/pathology , Neoplasm Recurrence, Local , Pancreas/surgery , Pancreatitis, Chronic/surgery , Retrospective Studies , Postoperative Complications/surgery
2.
Am Surg ; 89(12): 6348-6350, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37159926

ABSTRACT

Pancreatoduodenectomy remains a complex abdominal operation for hpb surgeons. Significant complications keep on occurring to many patients undergoing Whipple procedure. We present ten patients, who required completion pancreatectomy in the early postoperative period after Whipples procedure, due to postoperative complications. Indications for completion pancreatectomy included: Sepsis secondary to uncontrolled GRADE C postoperative pancreatic fistula, pancreatic leak and bleeding, postoperative hemorrhage, pancreatic leak with gastrointestinal anastomosis dehiscence, and hepaticojejunal anastomosis dehiscence combined with hemorrhage. Completion pancreatectomy was carried out at a mean interval of 9 days following Whipple procedure. Six patients (60%) survived the operation and discharged from the hospital, with a median survival of 21.3 months. Four patients (40%) died in the early post-operative period due to sepsis (10%) and multiple organ failure (30%). Completion pancreatectomy after pancreatoduodenectomy is rarely indicated and it can be considered as a salvage procedure in the management of severe life-threatening post pancreatic surgery complications.


Subject(s)
Pancreatectomy , Sepsis , Humans , Pancreatectomy/methods , Pancreaticoduodenectomy/adverse effects , Pancreaticoduodenectomy/methods , Retrospective Studies , Pancreas/surgery , Postoperative Complications/etiology , Pancreatic Fistula/etiology , Pancreatic Fistula/surgery , Sepsis/etiology
3.
Cardiooncology ; 8(1): 16, 2022 Sep 07.
Article in English | MEDLINE | ID: mdl-36071532

ABSTRACT

Breast cancer patients are at a particularly high risk of cardiotoxicity from chemotherapy having a detrimental effect on quality-of-life parameters and increasing the risk of mortality. Prognostic biomarkers would allow the management of therapies to mitigate the risks of cardiotoxicity in vulnerable patients and a key potential candidate for such biomarkers are microRNAs (miRNA). miRNAs are post-transcriptional regulators of gene expression which can also be released into the circulatory system and have been associated with the progression of many chronic diseases including many types of cancer. In this review, the evidence for the potential application of miRNAs as biomarkers for chemotherapy-induced cardiotoxicity (CIC) in breast cancer patientsis evaluated and a simple meta-analysis is performed to confirm the replication status of each reported miRNA. Further selection of miRNAs is performed by reviewing the reported associations of each miRNA with other cardiovascular conditions. Based on this research, the most representative panels targeting specific chemotherapy agents and treatment regimens are suggested, that contain several informative miRNAs, including both general markers of cardiac damage as well as those for the specific cancer treatments.

4.
J Surg Case Rep ; 2022(5): rjac234, 2022 May.
Article in English | MEDLINE | ID: mdl-35665394

ABSTRACT

We report the case of a single 46-year-old woman presenting with huge uterine fibroids growing for the last 12 years, resulting in a recent common iliac vein thrombosis. Due to the high risk for pulmonary embolism, an occluding balloon was inserted through the right jugular vein before the abdominal incision and occluded the vena cava just inferior to the renal veins. The tumor was easily mobilized, and the vena cava bifurcation was exposed and controlled until the uterus with the masses was resected. We recommend this method for oncovascular surgeries involving deep vein thrombosis and vein thromboembolism.

6.
Shock ; 50(2): 199-208, 2018 08.
Article in English | MEDLINE | ID: mdl-28957875

ABSTRACT

The present study focuses on the profile of "endogeneous" caveolin-1 protein in septic lung (CLP model).Caveolin-1, CD25, pP38, pAkt, and 14-3-3b protein expression profiles were studied using flow cytometry and immunohistochemistry 6, 12, 24, 36, and 48 h after sepsis induction. Cell viability was determined by 7-AAD staining and fibrosis by Masson trichrome stain. The effect of protein C zymogen concentrate (PC) on caveolin-1 expression was also investigated given that PC, once dissociated from caveolin-1, elicits a PAR-1-mediated protective signaling by forming a complex with endothelial protein C receptor (EPCR).CLP treatment increased lung inflammation and cell apoptosis. Fibrosis was apparent in vessels and alveoli. Caveolin-1+ cells presented reduced protein expression, especially 12 h post-CLP (P = 0.002). Immunohistochemistry revealed caveolin-1 positive expression mainly in regions with strong inflammatory reaction. Early induction of pP38+ cell population (P = 0.014) and gradual increase of CD25+ cells were also observed. Alternations in 14-3-3b expression related to apoptosis were apparent and accompanied by increased AKT phosphorylation activity late during sepsis progression.After PC administration, cell apoptosis was reduced (P = 0.004) and both the percentile and expression intensity of caveolin-1 positive cells were compromised (P = 0.009 and P = 0.027, respectively). 14-3-3b, CD25, and pP38 protein expression were decreased (P = 0.014, P = 0.004, and P = 0.007, respectively), whereas pAkt expression was induced (P = 0.032).The observed decline of endogenous caveolin-1 protein expression during sepsis implies its involvement in host's cytoprotective reaction either directly, by controlling caveolae population to decrease bacterial burden, or indirectly via regulating 14-3-3b-dependent apoptosis and EPCR-PAR-1-dependent protective signaling.


Subject(s)
Apoptosis , Caveolin 1/biosynthesis , Cytoprotection , Down-Regulation , Lung/metabolism , Sepsis/metabolism , Signal Transduction , Animals , Disease Models, Animal , Lung/pathology , Protein C/pharmacology , Rats , Rats, Wistar , Sepsis/pathology
7.
Ann Gastroenterol ; 29(1): 56-62, 2016.
Article in English | MEDLINE | ID: mdl-26752951

ABSTRACT

BACKGROUND: Synbiotics (combination of prebiotics and probiotics) may serve as a supportive dietary supplement-based strategy after colectomy for cancer. The potential benefits of early postoperative administration of synbiotics on the gastrointestinal function-related quality of life inpatients were explored. METHODS: Patients who underwent elective colectomy were prospectively enrolled and randomized to receive either synbiotics (n=38) or placebo (n=37) on the day they tolerated liquid diet and for 15 days thereafter. Primary endpoints were Gastro-Intestinal Quality of Life Index (GIQLI) questionnaire assessments at 1, 3 and 6 months postoperatively. Secondary endpoints were functional bowel disorders ("diarrhea", "constipation") assessed by EORTC QLQ-C30. RESULTS: Patients under synbiotics had a better GIQLI "Global score" compared with those who received placebo [77±1.67 vs. 71.36±1.69, P=0.01 (1 month); 77±1.7 vs. 72.5±1.73, P=0.03 (3 months); 79.23±1.82 vs. 72.75±1.85, P=0.01 (6 months)]. Multivariate linear mixed model analysis showed that synbiotics administration was the only independent significant factor for the "Global score" amelioration (b: 5.42, SE (b)1.8, 95%CI 1.78-9.1, P=0.004). The EORTC QLQ-C30 "diarrhea" domain score differences from baseline were better after synbiotics administration after 3 (P=0.04) and 6 months (P=0.003). No significant effect on "constipation" scores was observed. CONCLUSION: Synbiotics administration may have a beneficial effect on the postcolectomy gastrointestinal function.

8.
Shock ; 45(2): 139-47, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26513702

ABSTRACT

The impact of a potential autophagy (LC3a/b) deregulation in hyper and in hypo stages during sepsis-induced kidney injury and the temporal profile of phosphorylated extracellular signal-related kinase, P38 (pP38), Akt (pAKT), and 13-3-3ß protein were investigated in the current study, using a rat cecal ligation and puncture (CLP) model, by means of flow cytometry and immunohistochemistry. Cell viability was assessed by protein C zymogen concentrate (PC), 7-aminoactinomycin D (7-AAD) staining and inflammation by S100 protein immunostaining. The impact of reduced kidney inflammation in autophagy was assessed by PC administration, an anti-inflammatory and cytoprotective substance. Sepsis induction increased LC3a/b expression, which presented two peaks at 6 and 36 h after CLP, both in the percentage of positive cells (P = 0.024, P = 0.025, respectively) and in fluorescence intensity. At 6 h when inflammation was already apparent, LC3a/b increase was escorted by phosphorylated extracellular signal-related kinase stimulation and high cell viability (65%), designating autophagy as a cytoprotective mechanism against microbial infection. The phosphorylation of P38 was delayed to 12 h after CLP, when autophagy was reduced. pAkt and 14-3-3ß expression was stimulated between 6 and 36 h after CLP, although a slight inhibition of pAkt within each cell was detected (lower MnIX value). During the second peak, inflammation was intensified, necrosis was significantly increased with LC3a/b+/7-AAD + cells to present a 1.5-fold increase. Protein C zymogen concentrate administration declined autophagy at 6 and 36 h after CLP and reduced necrosis, whereas double positive LC3a/b and 7-AAD cells were increased by 1.68 and 2.78-fold, respectively. These data open new prospectives in sepsis treatment, since they further support that autophagy represents a cytoprotective mechanism triggered by stress conditions, rather than an alternative cell death pathway.


Subject(s)
Acute Kidney Injury/physiopathology , Autophagy/physiology , Inflammation/physiopathology , Sepsis/physiopathology , Animals , Apoptosis/physiology , Cecum/physiopathology , Cells, Cultured , Dactinomycin/analogs & derivatives , Dactinomycin/metabolism , Flow Cytometry , Immunohistochemistry , Ligation , Male , Models, Theoretical , Rats , Rats, Wistar , Signal Transduction/physiology
9.
Surg Endosc ; 28(3): 1027-33, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24185751

ABSTRACT

BACKGROUND: Global rise in the incidence of obesity and type 2 diabetes mellitus is widely recognized as one of the most challenging contemporary threats to public health. Weight loss surgery has proven to be an effective and durable solution for morbidly obese adults. Laparoscopic sleeve gastrectomy (LSG) was introduced as a restrictive procedure for obese patients, initially described as a possible first-stage operation, but now commonly performed as a stand-alone bariatric operation for both high-risk and super-morbid-obese patients, as well as for patients with lower body mass index. This study aims to evaluate the progression of glucose metabolism in patients undergoing LSG. METHODS: This prospective study investigated 62 patients who underwent LSG by the same surgical team in an 18-month period. Preoperative evaluation included demographic information, complete medical history including comorbidities and medication, clinical examination, evaluation of cardiopulmonary function, measurement of weight and height on a standard electronic scale, upper gastrointestinal endoscopy and upper abdominal ultrasound, as well as interviews with a psychologist and nutritionist. Glucose metabolism was evaluated by oral glucose tolerance test (OGTT), preoperatively and at 3, 6, and 12 months after surgery. RESULTS: The OGTT was significantly ameliorated in all groups during follow-up. Nine of 12 diabetic patients (75 %) ceased drug treatment at 3 months postoperatively (p = 0.004), increasing to 100 % at 1-year follow-up (p < 0.001). Normoglycemic patients and patients with borderline OGTT experienced mild or severe hypoglycemia during the glucose tolerance test at 3, 6, and 12 months' follow-up. CONCLUSIONS: LSG offers excellent results to morbidly obese patients with regard to type 2 diabetes mellitus. Implementation of OGTT in these patients can be a valuable tool in their postoperative management. Bariatric teams performing LSG for morbid obesity should heighten their sensitivity to postoperative hypoglycemia, even in patients with type 2 diabetes mellitus.


Subject(s)
Blood Glucose/metabolism , Diabetes Mellitus, Type 2/blood , Gastrectomy/methods , Gastroplasty/methods , Laparoscopy , Obesity, Morbid/surgery , Adult , Body Mass Index , Diabetes Mellitus, Type 2/complications , Female , Follow-Up Studies , Glucose Tolerance Test , Humans , Male , Obesity, Morbid/blood , Obesity, Morbid/complications , Postoperative Period , Prospective Studies , Time Factors , Treatment Outcome
10.
J Neurosci Res ; 89(9): 1409-18, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21618583

ABSTRACT

The 14-3-3 family members play a crucial role in the determination of cell fate, exerting their antiapoptotic activity through directly interfering with the critical function of the mitochondrial core proapoptotic machinery. Dimerization of 14-3-3 is vital for the interaction with many of its client proteins and is regulated by phosphorylation. In a previous study, we observed time-dependent neuronal apoptosis during sepsis. Therefore, in the present study, we sought to evaluate the expression of 14-3-3 θ and ß isoforms in septic brain and their association with apoptosis. Sepsis was induced by a CLP model in Wistar rats that were sacrificed at predefined time points. Flow cytometric analysis showed a sepsis-induced, time-dependent alteration of 14-3-3 θ and ß isoforms in both Neun(+) and GFAP(+) cells. 14-3-3 θ was linearly correlated with apoptosis, and stratified analysis for alive and apoptotic neuronal cells demonstrated a gradual down-regulation of θ isoform in alive neurons and astrocytes. The phospho-P38 (pP38) MAP kinase levels were altered in a time-dependent manner during sepsis, presenting a peak at 6 hr post-CLP. A significant correlation between the two isoforms of 14-3-3 was observed in septic rats, with the θ isoform predominant at all time points. The hippocampus, Purkinje cells, and glia-like cells showed intense immunohistochemical reactivity for 14-3-3 θ isoform, whereas the choroid plexus showed constantly increased ß isoform expression. Our results showed that sepsis alters the expression of both 14-3-3 θ and ß isoforms in a time-, cell-, and topography-dependent manner.


Subject(s)
14-3-3 Proteins/metabolism , Apoptosis/physiology , Brain/metabolism , Neurons/metabolism , Sepsis/metabolism , Animals , Brain/pathology , Cecum/surgery , Disease Models, Animal , Male , Neurons/pathology , Protein Isoforms , Rats , Rats, Wistar , Time Factors , Tissue Distribution
11.
Surgery ; 149(3): 394-403, 2011 Mar.
Article in English | MEDLINE | ID: mdl-20869092

ABSTRACT

BACKGROUND: To our knowledge, the predictive value of procalcitonin for bowel strangulation has been evaluated in only 2 experimental studies that had conflicting results. The objective of this study was to evaluate the value of procalcitonin for early diagnosis of intestinal ischemia and necrosis in acute bowel obstruction. METHODS: We performed a prospective study of 242 patients with small- or large-bowel obstructions in 2005. A total of 100 patients who underwent operation were divided into groups according to the presence of ischemia (reversible and irreversible) and necrosis, respectively, as follows: ischemia (n = 35) and nonischemia groups (n = 65) and necrosis (n = 22) and nonnecrosis groups (n = 78). Data analyzed included age, sex, vital signs, symptoms, clinical findings, white blood cell count, base deficit, metabolic acidosis, procalcitonin levels on presentation, the time between symptom onset and arrival at the emergency department and the time between arrival and operation, and the cause of the obstruction. RESULTS: Procalcitonin levels were greater in the ischemia than the nonischemia group (9.62 vs 0.30 ng/mL; P = .0001) and in the necrosis than the non-necrosis group (14.53 vs 0.32 ng/mL; P = .0001). Multivariate analysis identified procalcitonin as an independent predictor of ischemia (P = .009; odds ratio, 2.252; 95% confidence interval, 1.225-4.140) and necrosis (P = .005; odds ratio, 2.762; 95% confidence interval, 1.356-5.627). Using receiver operating characteristic (ROC) curve analysis, the area under the curve (AUC) of procalcitonin for ischemia and necrosis was 0.77 and 0.87, respectively. A high negative predictive value for ischemia and necrosis of procalcitonin levels <0.25 ng/mL (83% and 95%, respectively) and a positive predictive value of procalcitonin >1 ng/mL were identified (95% and 90%, respectively). CONCLUSION: Procalcitonin on presentation is very useful for the diagnosis or exclusion of intestinal ischemia and necrosis in acute bowel obstruction and could serve as an additional diagnostic tool to improve clinical decision-making.


Subject(s)
Calcitonin/blood , Intestinal Obstruction/blood , Intestines/blood supply , Intestines/pathology , Ischemia/diagnosis , Protein Precursors/blood , Adult , Aged , Area Under Curve , Calcitonin Gene-Related Peptide , Female , Humans , Male , Middle Aged , Multivariate Analysis , Necrosis , Predictive Value of Tests , Prospective Studies
12.
Surgery ; 149(3): 411-5, 2011 Mar.
Article in English | MEDLINE | ID: mdl-20850853

ABSTRACT

BACKGROUND: Despite their safety and effectiveness in thyroid surgery, the previous harmonic scalpel instruments are considered large and cumbersome by several surgeons. An innovative technical improvement of the device has been made available since 2008. The objective of this study was to compare the results of total thyroidectomy using the new harmonic scalpel (FOCUS) with that with the previously available device (HARMONIC ACE). METHODS: A prospective randomized study of all total thyroidectomies between February and July 2008 was conducted. Patients (n = 90) were randomized to undergo total thyroidectomy with FOCUS (group A, n = 45) or HARMONIC ACE (group B, n = 45). RESULTS: No significant differences were identified between the 2 groups in terms of demographics, reoperative thyroid surgery, thyroid gland weight and diameter, pathologic diagnosis, preoperative and postoperative serum PTH and calcium levels, postoperative complications, duration of hospital stay, and final outcome. The mean operative time was less in group A than group B (63 ± 7 min vs 76 ± 8 min, P = .009). CONCLUSION: The new harmonic scalpel is a useful adjunct to the armamentarium of the thyroid surgeon. It is safe, effective, and hand friendly, offering great capabilities for delicate tissue grasping and dissection. Use of this device decreased operative time compared with the previously available instrument.


Subject(s)
Surgical Instruments , Thyroidectomy/instrumentation , Adult , Aged , Female , Humans , Male , Middle Aged , Prospective Studies
13.
Crit Care Med ; 38(1): 209-16, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19726977

ABSTRACT

OBJECTIVE: Study the effect of human protein C (PC) concentrate administration on organ damage and survival in septic rats. DESIGN: Animal study. SETTING: University laboratory. SUBJECTS: Male Wistar rats. INTERVENTIONS: Cecal ligation and puncture (CLP) was performed in 210 rats. Rats were randomly assigned to receive either human protein C (PC) IV 1, 7, and 13 hrs after CLP (CLP+PC) or placebo (CLP). Septic animals were again randomized in a survival group (CLP: n = 50 and CLP+PC: n = 40) that was monitored for 60 hrs and time groups (CLP: n = 60 and CLP+PC: n = 60) that were killed at 6, 12, 24, 36, 48, and 60 hrs after CLP. Brain, heart, lung, liver, kidney, gastric, and colon tissue were removed and postfixed in paraffin sections. MEASUREMENTS AND MAIN RESULTS: PC infusion increased PC serum levels in early sepsis (median 7.25) compared with late sepsis (median 2, p = .001). Activated protein C/a1-antitrypsin complex levels in the CLP+PC group were significantly increased in late sepsis (60 hrs after CLP) compared with early sepsis (6, 12, and 24 hrs after CLP, p = .009, p = .004, and p = .008, respectively) and to late septic CLP and normal rats (p = .005 and p = .007, respectively). Their IL-6 and tumor necrosis factor a plasma levels were decreased (by 27% and 25%, respectively) at 6 hrs compared with placebo (p = .008 and p = .016). Their serum PC levels were also decreased in CLP+PC survivors compared with nonsurvivors of the same group (median = 1.5 vs. median = 7, p = .001). Apoptosis was reduced in brain (10% vs. 77.8%, p < .001), stomach (66.7% vs. 100%, p < .002) and intestine (33.3% vs. 85.2%, p < .001) compared with placebo. Finally, the survival of septic rats treated with human PC was significantly increased compared with placebo (75% vs. 54%, p = .033). CONCLUSIONS: Human Protein C administration increased survival in septic rats, decreased plasma inflammatory cytokines levels and tissue injury in vital organs.


Subject(s)
Cytokines/blood , Protein C/pharmacology , Sepsis/drug therapy , Sepsis/mortality , Animals , Cecum/surgery , Chi-Square Distribution , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Humans , Immunohistochemistry , In Situ Nick-End Labeling , Kaplan-Meier Estimate , Ligation , Male , Probability , Random Allocation , Rats , Rats, Wistar , Sepsis/pathology , Statistics, Nonparametric , Survival Rate
14.
J Surg Res ; 155(2): 183-90, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19645077

ABSTRACT

BACKGROUND: The detection of epithelial-specific mRNA correlates well with the presence of cancer cells in the peripheral blood and provides a rational explanation for subsequent metastasis. MATERIAL AND METHODS: Forty-two, patients with colorectal cancer and 14 controls were included in our study. Peripheral blood samples were acquired at 24 h before and 48 h after laparotomy. Tissue samples were also acquired from the primary lesion. All samples were examined for the expression profile of CEA, CK20, and TEM-8. RESULTS: Tissue samples expressed CEA in every specimen, CK20 in 30, and TEM-8 in 41. CEA and CK20 were not identified in the control blood samples while TEM-8 was detected in 4. CEA was detected in 17, CK20 in 28 and TEM-8 in 23, of the preoperative blood samples. CEA mRNA expression in preoperative blood sample and TNM stage were found independently associated with increased tumor size. Positive CEA, CK20, and TEM-8 signals were found in 25, 25, and 23 of the postoperative blood samples respectively. CONCLUSIONS: CK20 and CEA are significantly more frequently detected in colon cancer patients than in healthy controls and can serve as markers. Cancer cell mRNA is commonly detected in the preoperative and postoperative peripheral blood samples. Tumor size was independently associated with the preoperative detection of CEA mRNA. Although TEM-8 mRNA detection in the peripheral blood showed no specificity for cancer patients or correlation with clinical stage, identification and validation of genes and proteins implicated in metastatic process needs to be further investigated.


Subject(s)
Biomarkers, Tumor/metabolism , Carcinoembryonic Antigen/metabolism , Colorectal Neoplasms/metabolism , Epithelial Cells/metabolism , Keratin-20/metabolism , Neoplasm Proteins/metabolism , Receptors, Cell Surface/metabolism , Aged , Carcinoembryonic Antigen/genetics , Case-Control Studies , Centrifugation, Density Gradient/methods , Colorectal Neoplasms/pathology , Epithelial Cells/pathology , Feasibility Studies , Female , Humans , Keratin-20/genetics , Male , Microfilament Proteins , Middle Aged , Neoplasm Proteins/genetics , RNA, Messenger/blood , Receptors, Cell Surface/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Sensitivity and Specificity
15.
Brain Res ; 1264: 119-26, 2009 Apr 06.
Article in English | MEDLINE | ID: mdl-19368819

ABSTRACT

Activated Protein C renders anti-apoptotic properties in neurons and endothelial cells. The aim of the present study was to evaluate the in vivo cytoprotective role of Protein C zymogen (PC) administration in septic rat brain. Male Wistar rats (n=60) were subjected to sepsis via Cecal Ligation and Puncture (CLP). Animals were randomly divided either to receive 100 IU/kg human PC concentrate at 1, 7 and 13 h post CLP (CLP+PC group) or placebo treatment (CLP group). At pre-specified time points (6, 12, 24, 36, 48 and 60 h post CLP) five animals from either group were euthanized and the brain tissue was removed. Apoptosis in both neurons (Neu-N+) and astroglia (GFAP+) was assessed by flow cytometry using 7-aminoactinomycin D (7AAD). Immunohistochemical detection of cleaved caspase 3, bax, bcl-2, cytochrome c and caspase 8 was also performed. PC treated animals had significantly reduced apoptosis in neurons at 6 and 24 h post CLP (p=0.04 and p=0.016 respectively) and necrosis at 6, 12 and 60 h post CLP (p=0.008, p=0.012 and p=0.032 respectively). Astrocyte necrosis was also decreased in septic rats receiving PC (6, 12 and 60 h post CLP p=0.008, p=0.016 and p=0.008 respectively). In addition, active caspase 3, bax, cytochrome c and caspase 8 expression was significantly decreased during early sepsis (6-36 h) while bcl-2 expression was increased (24 h p=0.001 and 60 h p=0.001) in the PC treated animals compared to placebo. PC concentrate administration in experimental sepsis produced a time dependent inhibition of apoptosis in rat neurons and astrocytes. The inhibition of sepsis related apoptosis concerned both the mitochondrial and caspase 8 dependent pathways.


Subject(s)
Apoptosis/drug effects , Brain/drug effects , Cytoprotection , Protein C/pharmacology , Sepsis/pathology , Animals , Astrocytes/drug effects , Astrocytes/metabolism , Astrocytes/pathology , Brain/metabolism , Brain/pathology , Caspases/metabolism , Cytochromes c/metabolism , Flow Cytometry , Immunohistochemistry , Male , Neurons/drug effects , Neurons/metabolism , Neurons/pathology , Proto-Oncogene Proteins c-bcl-2/metabolism , Rats , Rats, Wistar , Sepsis/metabolism , Time Factors , bcl-2-Associated X Protein/metabolism
17.
Surg Infect (Larchmt) ; 9(3): 377-88, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18570579

ABSTRACT

BACKGROUND AND PURPOSE: Renal dysfunction attributable to sepsis was long considered a result of hemodynamic instability and subsequent local ischemia. Recent data show that apoptosis may be implicated also. The purpose of this study was to evaluate the role of apoptosis and the expression of the bax, bcl-2, caspase-8, and cytochrome c proteins in the renal parenchymal cells of rats with sepsis. METHODS: Sepsis was induced using cecal ligation and puncture (CLP) in 62 male Wistar rats, which were euthanized 6, 12, 24, 36, 48, or 60 h later. Ten sham-treated animals served as a control group. Another group of 50 animals were subjected to CLP and then supervised for 60 h. Renal apoptosis was evaluated using light and transmission electron microscopy, in situ nick-end labeling (TUNEL), and flow cytometry using 7-amino-actinomycin D (7-AAD). Caspase-mediated apoptosis was assessed using M30 antibody. The expression of the apoptosis-regulator proteins B-cell lymphoma 2 (bcl-2), bcl-2-associated x protein (bax), caspase-8, and cytochrome c was detected immunohistochemically. RESULTS: Sepsis increased inflammatory infiltration (p < 0.001) and necrosis (p < 0.001) in renal parenchyma. Apoptosis was significantly more common than in the kidneys of control animals (p = 0.02). Nuclei stained by the TUNEL technique were predominant in the tubular cells of non-survivors (p = 0.05). The time distribution of all types of cell death was increased significantly 6 h after the induction of sepsis, and declined subsequently. Caspase-generated cytokeratin 18 (CK18) new epitope (M30) was significantly more abundant in the kidneys of animals with sepsis than in control rats, with peaks at 6 h and 60 h post-procedure (p < 0.001). In addition, cells initiating apoptosis were significantly more common at 6 h than at 48 h post-CLP (p = 0.014). Caspase-8 protein immunodetection followed the same time pattern as cell death, increasing as early as 6 h post-CLP and decreasing thereafter (p = 0.013). Bax protein expression was elevated significantly early in the course of sepsis (p = 0.037), whereas the other members of the mitochondrial-dependent pathway remained constant. Animals dying from sepsis had a significantly greater prevalence of bax- (p = 0.037) and caspase-8- (p = 0.031) immunoreactive renal cells. CONCLUSION: Apoptosis in renal tissue was significantly more common in animals with sepsis than in controls. The time distribution of cell death markers showed a consistent pattern, making early sepsis the likely initiator of the apoptotic events.


Subject(s)
Apoptosis , Kidney Tubules/cytology , Kidney Tubules/metabolism , Sepsis/physiopathology , Animals , Caspase 8/metabolism , Cytochromes c/metabolism , Flow Cytometry , Immunohistochemistry , In Situ Nick-End Labeling , Keratin-18/metabolism , Kidney Tubules/physiology , Male , Microscopy, Electron , Proto-Oncogene Proteins c-bcl-2/metabolism , Rats , Rats, Wistar , Sepsis/mortality , Sepsis/pathology , Specific Pathogen-Free Organisms , bcl-2-Associated X Protein/metabolism
18.
World J Gastroenterol ; 14(9): 1389-93, 2008 Mar 07.
Article in English | MEDLINE | ID: mdl-18322953

ABSTRACT

AIM: To determine end-stage pathologic changes in the liver of septic patients dying in the intensive care unit. METHODS: Needle liver biopsies obtained immediately after death from 15 consecutive patients with sepsis and no underlying liver disease were subjected to routine histological examination. Liver function tests and clinical monitoring measurements were also recorded. RESULTS: Liver biochemistries were increased in the majority of patients before death. Histology of liver biopsy specimens showed portal inflammation in 73.3%, centrilobular necrosis in 80%, lobular inflammation in 66.7%, hepatocellular apoptosis in 66.6% and cholangitis/cholangiolitis in 20% of patients. Mixed hepatitic/cholestatic type of liver injury was observed in 6/15 (40%) patients and hepatitc in 9/15 (60%). Steatosis was observed in 11/15 (73.3%) patients affecting 5%-80% of liver parenchyma. Among the histological features, the presence of portal inflammation in liver biopsy was associated with increased hospitalization in the ICU prior death (P=0.026). CONCLUSION: Features of hepatitis and steatosis are the main histological findings in the liver in the majority of patients dying from sepsis.


Subject(s)
Inpatients , Intensive Care Units , Liver/microbiology , Liver/pathology , Sepsis/microbiology , Sepsis/pathology , Adult , Aged , Aged, 80 and over , Apoptosis , Biopsy, Needle , Fatty Liver/microbiology , Fatty Liver/pathology , Female , Hepatitis/microbiology , Hepatitis/pathology , Humans , Male , Middle Aged
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