Postoperative Blood Samples Research Articles

Long-term safety from transmission of porcine endogenous retrovirus after pig-to-non-human primate corneal transplantation.

The risk of xenozoonosis mainly by porcine endogenous retrovirus (PERV) has been considered as one of the main hurdles in xenotransplantation and therefore should be elucidated prior to the clinical use of porcine corneal grafts. Accordingly, an investigation was performed to analyze the infectivity of PERVs from porcine keratocytes to human cells, and the long-term risk of transmission of PERVs was determined using pig-to-non-human primate (NHP) corneal transplantation models. The infectivity of PERVs from the SNU miniature pig keratocytes was investigated by coculture with a human embryonic kidney cell line. Twenty-two rhesus macaques underwent xenocorneal transplantation as follows: (i) group 1 (n=4): anterior lamellar keratoplasty (LKP) with freshly preserved porcine corneas, (ii) group 2 (n=5): anterior LKP with decellularized porcine corneas followed by penetrating keratoplasty (PKP) with allografts, (iii) group 3 (n=3): PKP under steroid-based immunosuppression, (iv) group 4 (n=4): PKP under anti-CD154 antibody-based immunosuppression, (v) group 5 (n=4): deep anterior LKP with freshly preserved porcine corneas under anti-CD40 antibody-based immunosuppression, and (vi) group 6 (n=2): PKP under anti-CD40 antibody-based immunosuppression. Postoperative blood samples were serially collected, and tissue samples were obtained from thirteen different organs at the end of each experiment. The existence of PERV DNA and RNA was investigated using PCR and RT-PCR. Using two independent in vitro infectivity tests, neither PERV pol nor pig mitochondrial cytochrome oxidase II was detected after 41 and 92days of coculture, respectively. After xenocorneal transplantation, a total of 257 serial peripheral blood mononuclear cell samples, 34 serial plasma samples, and 282 tissue samples were obtained from the NHP recipients up to 1176days post-transplantation. No PERV transmission was evident in any samples. Within the limits of this study, there is no evidence to support any risk of PERV transmission from porcine corneal tissues to NHP recipients, despite the existence of PERV-expressing cells in porcine corneas.

Peripheral blood leucocytes show differential expression of tumour progression-related genes in colorectal cancer patients who have a postoperative intra-abdominal infection: a prospective matched cohort study.

Anastomotic leak is associated with higher rates of recurrence after surgery for colorectal cancer. However, the mechanisms responsible are unknown. We hypothesized that the infection-induced inflammatory response may induce overexpression of tumour progression-related genes in immune cells. The aim was to investigate the effect of postoperative intra-abdominal infection on the gene expression patterns of peripheral blood leucocytes (PBL) after surgery for colorectal cancer. Prospective matched cohort study. Patients undergoing surgery for colorectal cancer were included. Patients who had anastomotic leak or intra-abdominal abscess were included in the infection group (n=23) and matched with patients without complications for the control group (n=23). PBL were isolated from postoperative blood samples. Total RNA was extracted and hybridized to the Affymetrix Human Gene 1.0 ST microarray. Patients in the infection group displayed 162 upregulated genes and 146 downregulated genes with respect to the control group. Upregulated genes included examples coding for secreted cytokines involved in tumour growth and invasion (S100P, HGF, MMP8, MMP9, PDGFC, IL1R2). Infection also upregulated some proangiogenic genes (CEP55, TRPS1) and downregulated some inhibitors of angiogenesis (MME, ALOX15, CXCL10). Finally, some inhibitors (HP, ORM1, OLFM4, IRAK3) and activators (GNLY, PRF1, FGFBP2) of antitumour immunity were upregulated and downregulated, respectively, suggesting that the inflammatory environment caused by a postoperative infection favours immune evasion mechanisms of the tumour. Analysis of PBL shows differential expression of certain tumour progression-related genes in colorectal cancer patients who have a postoperative intra-abdominal infection, which in turn may promote the growth of residual cancer cells to become recurrent tumours.

Preoperative Tim‑3 expression on peripheral NK cells is correlated with pathologic TNM staging in colorectal cancer.

Previous research has indicated that T cell immunoglobulin and mucin domain 3 (Tim-3) serves an important regulatory role in lymphocytes and in several cancers. However, the association between Tim‑3 expression on various lymphocyte subsets and human colorectal cancer (CRC) has not been elucidated. The present study aimed to characterize Tim‑3 expression on peripheral lymphocytes, including cluster of differentiation CD3+CD56‑ T cells, CD3‑CD56+ natural killer (NK) cells and CD3+CD56+ natural killer T (NKT) cells, in patients with CRC. The frequency of T cells, NK cells and NKT cells expressing Tim‑3 was assessed by multicolor flow cytometry of peripheral blood collected from 36 preoperative CRC patients and 38 healthy donors. The expression of Tim‑3 on lymphocyte subsets from 53 postoperative blood samples of CRC patients was also analyzed. There were fewer circulating NK cells in patients with CRC compared with healthy controls (P=0.0027); NK cell expression of Tim‑3 was also significantly decreased (P=0.0239). The frequency of circulating NK cells and Tim‑3+ NK cells was negatively correlated with clinical cancer stage, compared with healthy controls, but not with other clinicopathological parameters or serum concentrations of CRC biomarkers. Furthermore, the expression of Tim‑3 in NK cells was higher in CRC patients without metastasis. Notably, NK cell Tim‑3 expression in CRC patients was significantly restored following surgical resection of the primary tumor. In conclusion, the present study indicates the presence of an altered frequency and expression of Tim‑3 in peripheral NK cells in CRC patients. Preoperative Tim‑3 expression on peripheral NK cells is correlated with differential staging in colorectal cancer, and may be useful as a serum biomarker.

The relationship between systemic inflammation and stoma formation following anterior resection for rectal cancer: A cross-sectional study

IntroductionThere is evidence that temporary defunctioning stoma formation in patients undergoing anterior resection reduces the risk of anastomotic leakage. The aim of the present study was to investigate the relationship between stoma formation, the postoperative systemic inflammatory response and complications following anterior resection for rectal cancer. MethodsData was recorded prospectively for patients who underwent anterior resection for histologically proven rectal cancer, from 2008 to 2015 at a single centre, n = 167. Patients had routine preoperative and postoperative blood sampling including serum C-reactive protein (CRP). Postoperative complications including anastomotic leakage were recorded. ResultsOf the 167 patients, the majority were male (61%) and over 65 years old (56%) with node negative disease (60%). 36 patients (22%) underwent preoperative neoadjuvant treatment. 100 patients (60%) had a stoma formed at the time of surgery. Stoma formation was significantly associated with male sex (69% vs. 50%, p = 0.017), neoadjuvant chemoradiotherapy (30% vs 9%, p = 0.001) and open surgery (71% vs. 55%, p = 0.040). Of those 100 patients who had a stoma formed, 80 had it reversed. Permanent stoma was significantly associated with increasing age (p = 0.011), exceeding the established CRP threshold of 150 mg/L on postoperative day 4 (67% vs 37%, p = 0.039), higher incidence of postoperative complications (76% vs 47%, p = 0.035), anastomotic leakage (24% vs 2%, p = 0.003) and higher Clavien Dindo score (p = 0.036). ConclusionsThere was no significant association between stoma formation during anterior resection and the postoperative systemic inflammatory response. However, in these patients both the postoperative systemic inflammatory response and complications were associated with permanent stoma.

Change in number and size of circulating tumor cells with high telomerase activity during treatment of patients with gastric cancer.

Detection of circulating tumor cells (CTCs) in peripheral blood is useful for estimating the prognosis of patients with cancer. We previously reported the detection of CTCs by OBP-401, a telomerase-specific, replication-selective, oncolytic adenoviral agent carrying the green fluorescent protein (GFP) gene. We demonstrated that the number of large (L)-GFP+ cells (≥7.735 µm in diameter) in peripheral blood samples correlated significantly with the prognosis of treatment-naïve gastric cancer patients, whereas the number of small (S)-GFP+ cells (<7.735 µm in diameter) did not. In the present study, we studied the change in the number of GFP+ cells during treatment, and analyzed the association between the number of GFP+ cells in blood samples and the outcome of patients. Peripheral blood samples were obtained from 37 gastric patients prior and subsequent to surgery (three samples per time point). Upon infection of blood cells with OBP-401, GFP+ cells of different sizes were counted and measured. The association between the number of GFP+ cells and surgical outcome was determined by statistical analysis. The median follow-up period after surgery was 39 months. Although the difference was not significant, patients with ≥6 L-GFP+ cells in preoperative blood samples had a lower relapse-free survival rate than patients with 0-5 L-GFP+ cells. There was no significant correlation between the number of L-GFP+ cells in postoperative blood samples and the prognosis of patients receiving adjuvant therapy. Although the difference was not significant, the number of S-GFP+ cells in samples from patients who had received postoperative chemotherapy was higher than in those who had not. The number of L-GFP+ cells was not significantly correlated with the relapse-free survival rate in gastric cancer patients who underwent surgery. The number of S-GFP+ cells was relatively high in samples from patients who had received postoperative chemotherapy.

Screen and identification of serum protein biomarkers in gastric cancer

To screen and identify the serum specific protein markers of patients with gastric cancer by proteomics technology, and to provide more comprehensive serum protein fingerprint model for the early diagnosis of gastric cancer. Preoperative and postoperative blood samples were collected from 60 gastric cancer patients. Mass spectrometry (SELDI-TOF-MS) technology was used to detect and screen serum specific proteins in gastric cancer patients(preoperative group, postoperative group, metastasis group), and the result was compared with normal control group. Gel electrophoresis(TRICINE SDS-OAGE) technology was applied in the separation and purification for those different protein. Matrix assisted laser desorption ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF) technology was used in the identification for the proteins following separation and purification. Mass spectrometry data of preoperative group and normal group resulted in 15 specific m/z peak(P<0.01). SVM screened by a combination of the highest index model Youden get m/z peak at 6 449.1 protein markers. The protein expression of preoperative group was significantly higher than that of normal group(2 299.3±2 029.3 vs. 509.5±168.3, P<0.01). Mass spectrometry data of preoperative group and postoperative group resulted in 6 specific m/z peak(P<0.01). SVM screened by a combination of the highest Youden index model indentified get m/z peak at 6 449.2 protein markers. The protein expression of preoperative group was significantly higher than that of postoperative group(1 247.9±685.0 vs. 476.5±157.8, P<0.01). Mass spectrometry data of preoperative group and metastasis group resulted in 12 specific m/z peak (P<0.01). SVM screened by a combination of the highest Youden index model indentified get m/z peak at 6 448.9 protein markers. The protein expression of metastasis group was higher than that of preoperative group(1 506.9±1 036.5 vs. 649.7±621.0). MALDI-TOF/TOF identified that the protein with m/z peak at 6 449 was Apo CIII(. Apo CIII( may be the specific serum protein marker of gastric cancer, which may provide a more comprehensive serum protein fingerprint model for the early diagnosis of gastric cancer and a new way for further research.

Clinical utility of circulating tumor DNA (ctDNA) in resectable pancreatic ductal adenocarcinoma (PDAC).

247 Background: ctDNA is emerging as a promising biomarker, with potential utility in screening, detecting minimal residual disease after curative resection and monitoring treatment response or resistance in advanced disease. Most PDAC studies to date have focused on identifying mutant KRAS ctDNA in metastatic disease. Here we perform sequential ctDNA quantification in patients (pts) with resectable PDAC using a novel and highly sensitive multiplex technology to explore the clinical utility of ctDNA as a diagnostic and prognostic biomarker. Methods: Banked plasma and tumor samples from 18 pts with resected PDAC were retrieved. Plasma samples were collected 0-28 days before, and 28-70 days after surgery. DNA was extracted using standard protocols and analyzed using the OnTarget system, which enriches for DNA molecules containing hot spot mutations prior to sequencing. A 96-plex panel that includes the most prevalent mutations in KRAS, PIK3CA and TP53 was used. Results: 16 pts (89%) had at least 1 mutation detected by OnTarget in the tumor sample, most frequently in KRAS codon 12 (n = 14). ctDNA was detected in the pre-operative blood sample in 7/16 pts with tumor mutations (sensitivity 44%) and 0/2 pts without detectable tumor mutations (specificity 100%). Of the 10 pts with available post-operative blood samples, 1 did not have a tumor mutation. 4 pts had detectable ctDNA, 3 of whom have recurred. In contrast, 0 of the 5 pts without detectable post-operative ctDNA have recurred. At median follow-up of 37 weeks, recurrence-free survival (RFS) was significantly longer in pts without detectable ctDNA after surgery (median not reached vs 9 weeks, p = 0.022). Of 11 plasma samples with detectable ctDNA, 3 harbored mutations that were not detected in the primary tumor, including 2 non-KRAS mutations (GNAS R201H and PIK3CA E542K). Conclusions: Pre-operative ctDNA has low sensitivity, suggesting limited utility in PDAC screening. RFS was significantly longer in pts without detectable post-operative ctDNA; however this analysis is limited by small numbers and short follow-up. Discordance in hot spot mutations detected in tumor and matched plasma was observed in 27% of samples, possibly related to intratumoral heterogeneity.

The relationship between systemic inflammation and postoperative outcomes following neoadjuvant chemoradiotherapy for rectal cancer.

682 Background: The magnitude of the systemic inflammatory response before and after surgery for rectal cancer is increasingly understood to have an impact on clinical outcomes. The aim of the present study was to examine the impact of neoadjuvant chemoradiotherapy (nCRT) on pre- and postoperative systemic inflammation and clinical outcomes in patients undergoing surgery for rectal cancer. Methods: Data was recorded prospectively for patients who underwent elective, potentially curative surgery for rectal cancer, from 2008 to 2015 at a single centre, n = 251. Patients had routine pre- and postoperative blood sampling. Results: Of the 251 patients, the majority were male (62%) and over 65 years old (57%) with node negative disease (66%). 85 patients (33%) were allocated to preoperative nCRT based on probable margin threatening disease at preoperative MRI. nCRT was significantly associated with a higher proportion of patients having neutrophil lymphocyte ratio (NLR) &gt; 5 (39% vs. 12%, p &lt; 0.001), and a modified Glasgow Prognostic Score (mGPS) of 2 (14% vs. 6%, p = 0.035) prior to surgery. There was no significant association between nCRT and the magnitude of the postoperative systemic inflammatory response or complications. Of the 85 patients who underwent nCRT, there was a small but significant reduction in CRP following treatment (3.25 mg/L vs. 4.1 mg/L, p = 0.007). Of the 16 patients who were systemically inflamed prior to nCRT, 9 patients achieved mGPS score 0 after treatment (p = 0.004). There was a significant association between having an mGPS score 1 or 2 both before and after nCRT and breaching established thresholds of CRP on postoperative days 2 (190mg/L, 86% vs. 33%, p = 0.002), 3 (170mg/L, 86% vs. 38%, p = 0.015) and 4 (145mg/L, 71% vs. 36%, p = 0.031). Conclusions: Allocation to nCRT was significantly associated with a systemic inflammatory response prior to surgery for rectal cancer. nCRT was significantly associated with attenuation of existing systemic inflammation. Such attenuation of the systemic inflammatory response may in part explain the efficacy of nCRT in patients with rectal cancer.

Hypoxia-induced inflammation and purinergic signaling in cross clamping the human aorta.

Open aortic surgery evokes a systemic inflammatory response and is associated with high morbidity and mortality. Purinergic signaling has been shown to be crucial for maintaining vascular integrity and attenuating inflammation related to hypoxia. The involvement of purinergic signaling in cross clamping of major human arteries is unknown. Our aim was to compare systemic inflammatory responses and hypoxia-induced purinergic signaling in patients undergoing either open infra-renal abdominal aortic repair or infra-inguinal revascularization. Pre- and 24 h post-operative blood samples were gathered from 6 patients undergoing aortic clamping and 6 similar patients undergoing common femoral artery cross-clamping. Using Biorad Multipex™ 21- and 27-panels 48 different cytokines, chemokines and growth factors were analyzed, in addition to circulating levels of ATP, ADP, CD39, CD73 and HIF-1α, and compared between the groups. Several inflammatory cytokines were elevated from baseline levels after aortic clamping, but not after femoral cross clamping. Most pronoun rises were seen in IL-6 (667 %, P = 0.016) and HGF (760 %, P = 0.016). HIF-1α values showed a steady increase after clamping of either artery unless the subject underwent blood transfusion. Despite an adequate increase in HIF-1α CD39 and CD73 activity decreased significantly after aortic clamping (P = 0.047 and P = 0.016, respectively). Aortic clamping is associated with a clear and strong systemic inflammatory response and impaired repair mechanisms in terms of purinergic signaling. Patients undergoing open aorta repair could benefit from pre-operative medical therapy, which enhances CD73 expression.Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-015-1651-x) contains supplementary material, which is available to authorized users.

Next-day Parathyroid Hormone as a Predictor of Post-thyroidectomy Hypocalcemia

ABSTRACT Aims Total thyroidectomy is significantly complicated by parathyroid dysfunction and hypocalcemia. These aspects impact the decision regarding the timing of discharge and quantum of calcium supplementation required. Therefore, we aimed at evaluating the accuracy of next-day parathyroid hormone (PTH) level as a predictor of post-thyroidectomy hypocalcemia. Secondly, we aimed at establishing our institution's postoperative PTH level, which can accurately predict the development of post-thyroidectomy hypocalcemia to help us ensure the safe and early discharge of patients. Materials and methods A prospective observational study of 50 continuous patients undergoing thyroidectomy was conducted at a tertiary hospital in South India. Postoperative blood samples were collected for estimation of PTH, calcium, albumin, and phosphorous. The data were collated and results analyzed using Stata I/C 10.1. Results A total of 30% (15/50) of the patients had postoperative hypocalcemia (serum calcium &lt;8 mg/dL). Postoperative PTH was low (&lt;8 pg/mL) in 40% (20/50) of patients. There was a significant association between PTH &lt; 8 pg/mL and the presence of postoperative hypocalcemia (p = 0.029). The area under the receiver operating characteristic (ROC) curve was 0.7, and a next-day PTH of &lt;6 pg/mL showed the highest sensitivity and specificity (83 and 60% respectively) for the development of postoperative hypocalcemia, with a positive predictive value (PPV) and negative predictive value (NPV) of 83 and 60 respectively. Conclusion The PTH assessment performed the day after surgery is an acceptable test to predict post-thyroidectomy hypocalcemia; PTH &lt;6 pg/mL can be used as our institution's cutoff value. Department protocols for calcium and vitamin D supplementation following total thyroidectomy may be formulated based on the appropriately timed local postoperative PTH value to assist safe and early discharge of patients. Clinical significance Discharge protocols for patients undergoing thyroidectomy may be formulated based on the postoperative PTH values, thus enabling safe and early discharge of patients. How to cite this article Cherian AJ, Ramakant P, Paul TV, Abraham DT, Paul MJ. Next-day Parathyroid Hormone as a Predictor of Post-thyroidectomy Hypocalcemia. World J Endoc Surg 2016;8(3):203-207.

Serological and viraemic status of human cytomegalovirus infection in patients with colorectal cancer is not correlated with viral replication and transcription in tumours.

Colorectal cancer (CRC) is amongst the leading causes of cancer-related mortality worldwide. Emerging evidence suggests that human cytomegalovirus (HCMV) exists in the tumour tissue of CRC and is associated with disease outcome. To study whether tumoral HCMV is related to viral reactivation in blood, tumour specimens and pre- and post-operative blood samples from CRC patients were collected prospectively. PCR and quantitative PCR were performed to detect HCMV DNA. HCMV IgG and IgM antibodies were measured using a microparticle enzyme immunoassay. Transcription of a spliced HCMV UL73 gene transcript was analysed by quantitative reverse transcription PCR. HCMV was detected in 42.2% (35/83) of the tumour samples, with a low median viral load (30.08, range 2.33-5704 copies per 500 ng genomic DNA). The vast majority (80/81, 98.8%) of the CRC patients were seropositive for HCMV IgG. HCMV DNA was positive in 11.3% (22/194) of the pre-operative and 8.9% (15/168) of the post-operative blood samples. However, presence of HCMV and its viral load in tumours were not associated with the detection or viral loads in blood samples. About 26.67% (8/30) of the HCMV-positive tumours with available RNA had detectable viral UL73 transcripts, whilst none of the blood samples were positive for viral RNA (P < 0.0001). Therefore, presence of HCMV in tumours does not correlate with the serological or viraemic status of CRC patients. Active viral gene transcription occurred in the tumour but not in the blood of CRC patients. HCMV reactivation in CRC patients is possibly due to virus-cancer interactions in the CRC tumour microenvironment.

Metabolic surgery and intestinal gene expression: Digestive tract and diabetes evolution considerations.

To investigate the effects of bariatric surgery on metabolic parameters, incretin hormone secretion, and duodenal and ileal mucosal gene expression. Nine patients with type 2 diabetes mellitus (T2DM), chronic serum hyperglycemia for more than 2 years, and a body mass index (BMI) of 30-35 kg/m(2) underwent metabolic surgery sleeve gastrectomy with transit bipartition between May 2011 and December 2011. Blood samples were collected pre and 3, 6 and 12 mo postsurgery. Duodenal and ileal mucosa samples were collected pre- and 3 mo postsurgery. Pre- and postoperative blood samples were collected in the fasting state before ingestion of a standard meal (520 kcal) and again 30, 60, 90, and 120 min after the meal to determine hemoglobin A1c (HbA1c) levels and the lipid profile, which consisted of triglyceride and total cholesterol levels. Intestinal gene expression of p53 and transforming growth factor (TGF)-β was analyzed using quantitative reverse-transcription PCR. Gastric inhibitory polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) were quantified using the enzyme-linked immunoassay method and analyzed pre- and postoperatively. Student's t test or repeated measurements analysis of variance with Bonferroni corrections were performed as appropriate. BMI values decreased by 15.7% within the initial 3 mo after surgery (31.29 ± 0.73 vs 26.398 ± 0.68, P < 0.05) and then stabilized at 22% at 6 mo postoperative, resulting in similar values 12 mo postoperatively (20-25 kg/m(2)). All of the patients experienced improved T2DM, with 7 patients (78%) achieving complete remission (HbA1c < 6.5%), and 2 patients (22%) achieving improved diabetes (HbA1c < 7.0% with or without the use of oral hypoglycemic agents). At 3 mo postoperatively, fasting plasma glucose had also decreased (59%) (269.55 ± 18.24 mg/dL vs 100.77 ± 3.13 mg/dL, P < 0.05) with no further significant changes at 6 or 12 mo postoperatively. In the first month postoperatively, there was a complete withdrawal of hypoglycemic medications in all patients, who were taking at least 2 hypoglycemic drugs preoperatively. GLP-1 levels significantly increased after surgery (149.96 ± 31.25 vs 220.23 ± 27.55) (P < 0.05), while GIP levels decreased but not significantly. p53 gene expression significantly increased in the duodenal mucosa (P < 0.05, 2.06 fold) whereas the tumor growth factor-β gene expression significantly increased (P < 0.05, 2.52 fold) in the ileal mucosa after surgery. Metabolic surgery ameliorated diabetes in all of the patients, accompanied by increased anti-proliferative intestinal gene expression in non-excluded segments of the intestine.

Compression anastomotic ring-locking procedure (CARP) is a safe and effective method for intestinal anastomoses following left-sided colonic resection.

Compression anastomotic ring-locking procedure (CARP) is a novel procedure for creating colonic anastomoses. The surgical procedure allows perioperative quantification of the compression pressure between the intestinal ends within the anastomosis and postoperative monitoring of the anastomotic integrity. We have recently shown that CARP is a safe and effective method for colonic anastomoses in pigs, and the purpose of the present study was to evaluate CARP for colonic anastomoses in humans. This is a prospective study on 25 patients undergoing elective left-sided colonic resection. Time for evacuation of the anastomotic rings, perioperative compression pressure, and adverse effects were recorded. Postoperative blood samples were collected daily, and flexible sigmoidoscopy was performed 8-12 weeks after surgery to examine the anastomoses. Fourteen out of 25 patients underwent CARP. CARP was not used in 11 patients due to advanced tumor disease (two cases) and size restrictions (nine cases). No case of anastomotic leakage, bowel obstruction, or stenosis formation was observed. No device-related perioperative adverse events were noted. The surgical device evacuated spontaneously in all patients by the natural route after a median of 10 days. Perioperative compression pressure ranged between 85 and 280 mBar (median 130 mBar). Flexible sigmoidoscopy revealed smooth anastomoses without signs of pathological inflammation or stenosis in all cases. Our results indicate that the novel suture-less CARP is a safe and effective method for creating colonic anastomoses. Further studies are warranted in larger patient populations to compare CARP head-on-head with stapled and/or hand-sewn colonic anastomoses.

Abstract ES8-2: Circulating plasma tumor DNA as biomarkers for breast cancer therapy

Abstract Currently, oncologists use prior clinical trials data to recommend therapies based upon features of the tumor and clinical stage of the patient. A limitation with this approach is that these data are averaged from large groups of patients that are then applied to each individual. This creates uncertainty as there is currently no reliable method to identify whether an individual patient truly is at risk for an unfavorable outcome and would benefit from a specific intervention. Recently, we and others have demonstrated the feasibility of detecting circulating plasma tumor DNA in metastatic breast cancer patients using digital PCR. We have now extended these studies to include early stage breast cancer patients using the second generation platform of droplet digital PCR (ddPCR). In this study the feasibility and accuracy of circulating plasma tumor DNA (ptDNA) detection was prospectively assessed by screening for the presence of tumor PIK3CA mutations in patients' plasma prior to and after breast surgery using ddPCR. We observed in early stage breast cancer patients that ddPCR could detect PIK3CA mutations preoperatively with 93.3% sensitivity (14 of 15 mutations detected) and 100% specificity (no false positives in 30 tumors). Moreover, 5 of 10 patients with PIK3CA mutation positive tumors continued to have mutant PIK3CA detected in their postoperative blood sample despite having no clinical evidence of disease. This work demonstrates that droplet digital PCR (ddPCR) can be used to accurately detect cancer DNA mutations in the blood of early stage breast cancer patients including those with minimal, clinically undetectable disease. Our results suggest that ddPCR provides an accurate and relatively non-invasive method for measuring residual disease and could be developed as a biomarker to identify early stage breast cancer patients at higher risk for recurrence to help make informed decisions regarding adjuvant therapies. Ongoing studies assessing mutational profiles from ptDNA from metastatic patients as well as patients undergoing neoadjuvant therapies will be discussed. Citation Format: Ben Ho Park. Circulating plasma tumor DNA as biomarkers for breast cancer therapy [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr ES8-2.

Postoperative Copeptin Concentration Predicts Diabetes Insipidus After Pituitary Surgery.

Copeptin is a stable surrogate marker of vasopressin release; the peptides are stoichiometrically secreted from the neurohypophysis due to elevated plasma osmolality or nonosmotic stress. We hypothesized that following stress from pituitary surgery, patients with neurohypophyseal damage and eventual diabetes insipidus (DI) would not exhibit the expected pronounced copeptin elevation. The objective was to evaluate copeptin's accuracy to predict DI following pituitary surgery. This was a prospective multicenter observational cohort study. Three Swiss or Canadian referral centers were used. Consecutive pituitary surgery patients were included. Copeptin was measured postoperatively daily until discharge. Logistic regression models and diagnostic performance measures were calculated to assess relationships of postoperative copeptin levels and DI. Of 205 patients, 50 (24.4%) developed postoperative DI. Post-surgically, median [25th-75th percentile] copeptin levels were significantly lower in patients developing DI vs those not showing this complication: 2.9 [1.9-7.9] pmol/L vs 10.8 [5.2-30.4] pmol/L; P < .001. Logistic regression analysis revealed strong association between postoperative copeptin concentrations and DI even after considering known predisposing factors for DI: adjusted odds ratio (95% confidence interval) 1.41 (1.16-1.73). DI was seen in 22/27 patients with copeptin <2.5 pmol/L (positive predictive value, 81%; specificity, 97%), but only 1/40 with copeptin >30 pmol/L (negative predictive value, 95%; sensitivity, 94%) on postoperative day 1. Lack of standardized DI diagnostic criteria; postoperative blood samples for copeptin obtained during everyday care vs at fixed time points. In patients undergoing pituitary procedures, low copeptin levels despite surgical stress reflect postoperative DI, whereas high levels virtually exclude it. Copeptin therefore may become a novel tool for early goal-directed management of postoperative DI.

Comparaison entre carbétocine et oxytocine en cours de césarienne dans la prévention des hémorragies du post-partum

ObjectivesThe aim of the study was to compare the effectiveness of Carbetocin versus Oxyotcin during caesarean section for preventing postpartum haemorrhage. MethodsProspective observational study (before/after design). Five hundred and forty patients who received an injection of Oxytocin were compared to 262 patients with single injection of 100 micrograms of Carbetocin. The primary outcome was to compare the differential hematocrit level between pre- and postoperative blood samples. The secondary outcome was to compare differential hemoglobin level and the use of complementary therapies for postpartum haemorrhage. ResultsWe did not find any difference between the Oxytocin and Carbetocin groups on differential hematocrit level. There was no difference between the groups regarding the use of additionnal therapies (Sulproston injections, blood transfusions and surgery methods). The rate of postpartum haemorrhage was similar in the two groups (18.7% vs 21.6%; P=0.33). We found a lower percentage of patients with differential of hemoglobin level between 2g/dL and 4g/dL in the Carbetocin group (6.5% vs 15.6%, P<0.001). The proportion of patients requiring intravenous iron administration was significantly lower in the Carbetocin group (6.8% vs 13.8%, P=0.0036) ConclusionAccording to the primary outcome, there is no difference in effectiveness between carbetocin and oxytocin. Carbetocin seems to reduce the need for postoperative intravenous iron injection.

Detection of circulating tumor cell subpopulations in patients with head and neck squamous cell carcinoma (HNSCC).

BackgroundSince image based diagnostic tools fail to detect early metastasis in head and neck squamous cell carcinoma (HNSCC) it is crucial to develop minimal invasive diagnostic methods. A promising approach is to identify and characterize circulating tumor cells (CTC) in the peripheral blood of HNSCC patients. In this pilot study, we assessed which non-hematopoietic cell types are identifiable and whether their numbers differ in pre- and postoperative blood samples.Methods20 ml citrated peripheral blood was taken from 10 HNSCC patients before and after curative resection. CTC were enriched using density gradient centrifugation. CTC presence was verified by multi-immunofluorescence staining against cytokeratin (CK; epithelial), N-cadherin (mesenchymal); CD133 (stem-cell), CD45 (hematopoietic) and DAPI (nucleus). Individual cell type profiles were analyzed.ResultsWe were able to detect cells with epithelial properties like CK+/N-cadherin−/CD45− and CK+/CD133−/CD45− as well as cells with mesenchymal features such as N-cadherin+/CK−/CD45− and cells with both characteristics like N-cadherin+/CK+/CD45−. We also observed cells showing stem cell-like features like CD133+/CK−/CD45− and cells with both epithelial and stem cell-like features such as CD133+/CK+/CD45−. The number of CK positive cells (p = 0.002), N-cadherin positive cells (p = 0.002) and CD133 positive cells (p = 0.01) decreased significantly after resection. Kaplan-Meier test showed that the survival was significantly shorter when N-cadherin+ cells were present after resection (p = 0.04; 474 vs. 235 days; [HR] = 3.1).ConclusionsThis is - to the best of our knowledge- the first pilot study identifying different CTC populations in peripheral blood of HNSCC patients and showing that these individual cell type profiles may have distinct clinical implications.

CD26-positive/CD326-negative circulating cancer cells as prognostic markers for colorectal cancer recurrence.

The present study evaluated the presence and clinical relevance of a cluster of differentiation (CD)26+/CD326− subset of circulating tumor cells (CTCs) in pre- and post-operative blood samples of colorectal cancer patients, who had undergone curative or palliative intervention, in order to find a novel prognostic factor for patient management and follow-up. In total, 80 colorectal cancer patients, along with 25 healthy volunteers were included. The easily transferable methodology of flow cytometry, along with multiparametric antibody staining were used to selectively evaluate CD26+/CD326− CTCs in the peripheral blood samples of colorectal cancer patients. The multiparametric selection allowed any enrichment methods to be avoided thus rendering the whole procedure suitable for clinical routine. The presence of CD26+/CD326− cells was higher in advanced Dukes’ stages and was significantly associated with poor survival and high recurrence rates. Relapsing and non-surviving patients showed the highest number of CD26+/CD326− CTCs. High pre-operative levels of CD26+/CD326− CTCs correctly predicted tumor relapse in 44.4% of the cases, while 69% of post-operative CD26+/CD326− CTC-positive patients experienced cancer recurrence, with a test accuracy of 88.8%. By contrast, post-operative CD26+/CD326− CTC-negative patients showed an increase in the three-year progression-free survival rate of 86%, along with a reduced risk of tumor relapse of >90%. In conclusion, CD26+/CD326− CTCs are an independent prognostic factor for tumor recurrence rate in multivariate analysis, suggesting that their evaluation could be an additional factor for colorectal cancer recurrence risk evaluation in patient management.

Bacteremia after piezocision

The aim of this study was to investigate the presence of transient bacteremia after a piezocision procedure. The sample consisted of 30 subjects (24 women, 6 men; mean age, 19.6±0.7years; range, 18.1-22.4years) with the American Society of Anesthesiologists' physical status I. All patients had Class I skeletal and dental relationships and had fixed orthodontic treatment with the Damon system. The piezocision surgery was performed 1week after the placement of the orthodontic appliances in all patients. Two 20-mL venous blood samples were collected before and 30 to 60seconds after the first microincision using an aseptic technique. The samples were inoculated into BACTEC Plus aerobic and anaerobic blood culture bottles and were assessed in the BACTEC blood culture analyzer (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md). The results were analyzed statistically using the McNemar test, with P<0.05 indicating statistical significance. No significant difference between the preoperative and postoperative samples was determined with respect to transient bacteremia (P=0.250). No bacteremia was detected in the pretreatment samples, although Gemella sanguinis, Streptococcus pluranimalium, and Streptococcus mitis/oralis were detected in 3 postoperative blood samples. The piezocision procedure might be related to transitory bacteremia. Hence, orthodontists should consider the possibility of bacterial endocarditis in at-risk patients when piezocision is part of the treatment plan.

Effects of diclofenac sodium on rat hippocampus exposed to acute unilateral subdural hematoma: A histological, stereological and molecular study

In this study, it was aimed to evaluate the potential effects of acute subdural hematoma (ASDH) and diclofenac sodium therapy following ASDH on the rat hippocampus by histological, stereological and biochemical methods. For this aim, 24 male Sprague-dawley rats were used. The subjects were divided into 4 groups. One ml venous blood was collected from the tail vein of the all animals before seven day from experimental acute subdural hematoma. Plasma serotonin, adrenaline and noradrenaline levels were measured in these plasma samples. During experimental acute subdural hematoma: Non-heparinized 0.1 ml of autologous blood from the tail vein of the animals in Group 3 and 4 were given to subdural space. Group 4 daily received intra-muscular diclofenac sodium administration at 15-mg/kg-dose from the postoperative second hour to 7 th day of the operation. Group 1 and 2 used for control and sham operation, respectively. At postoperative 8 th day, 1 ml venous blood from the all animals was collected before perfusion. The brain of animals were analysed by histological, stereological and biochemical techniques. The mean number of neurons in the CA1, CA2, CA3 regions of the hippocampus was estimated. Total number of neurons in the hippocampus of the diclofenac sodium treated group was significantly decreased. When comparing of postoperative blood samples, there was no difference between the levels of adrenaline and serotonin levels among the groups. But, only noradrenalin levels in the treated group it found to be higher than those of the sham and control groups (p<0.05). In this study, it was shown that using of diclofenac sodium after acute subdural hematoma caused to loss of the neurons in the hippocampal CA1, CA2, and CA3 regions.