Detection of cancer cells and tumor margins during colorectal cancer surgery by intraoperative flow cytometry.

BACKGROUND: Flow Cytometry is an analytical technique for the precise quantification of cellular phenotype. Intraoperative Flow Cytometry (iFC) utilizes flow cytometry for DNA content/ploidy and cell cycle distribution analysis during surgery for cancer cell characterization and evaluation of tumor margins. Various types of cancers, including intracranial, head and neck, breast and liver malignancies have been evaluated with iFC. In the current study we present an intraoperative Flow Cytometry protocol for colorectal cancer cell detection and potential resection margin evaluation. MATERIALS AND METHODS: This study includes 106 colorectal cancer patients in which samples from cancer and normal colon epithelium were prospectively collected intraoperatively and comparatively assessed with iFC. Patients' demographics, tumor data and cytometry parameters were assessed. RESULTS: We have demonstrated that a cut-off value of 10.5% for tumor-index (fraction of cells in S and G2/M cell cycle phases) predicts with ∼91% accuracy (82.2% sensitivity and 99.9% specificity) the presence of cancer cells. Evaluation of tumor margins by iFC in the subpopulation of rectal cancer patients with or without neoadjuvant therapy, revealed an accuracy of 79% and 88%, respectively. CONCLUSION: Our data support that regarding colorectal cancer, iFC is a useful adjunct method for tumor cell identification and probably margin evaluation, which could be utilized in rectal cancer treatment in the era of organ sparing procedures.

The Humoral Immune Response to BNT162b2 Vaccine Is Associated With Circulating CD19+ B Lymphocytes and the Naïve CD45RA to Memory CD45RO CD4+ T Helper Cells Ratio in Hemodialysis Patients and Kidney Transplant Recipients.

Background: The humoral and cellular immune responses to SARS-COV-2 vaccination remain to be elucidated in hemodialysis (HD) patients and kidney transplant recipients (KTRs), considering their baseline immunosuppressed status. The aim of our study was to assess the associations of vaccine-induced antibody responses with circulating lymphocytes sub-populations and their respective patterns of alterations in maintenance HD patients and KTRs. Materials and Methods: We included 34 HD patients and 54 KTRs who received two doses of the mRNA-vaccine BNT162b2. Lymphocyte subpopulations were analyzed by flow cytometry before vaccination (T0), before the second vaccine dose (T1) and 2 weeks after the second dose (T2). The anti-SARS-CoV2 antibody response was assessed at T1 and at T2. Results: 31 HD patients (91.8%) and 16 KTRs (29.6%) became seropositive at T2. HD patients who became seropositive following the first dose displayed higher CD19+ B lymphocytes compared to their seronegative HD counterparts. A positive correlation was established between CD19+ B cells counts and antibody titers at all time-points in both groups (p < 0.001). KTRs showed higher naïve CD4+CD45RA+ T helper cells compared to HD patients at baseline and T2 whereas HD patients displayed higher memory CD45RO+ T cells compared to KTRs at T2. The naïve CD4+CD45RA to memory CD4+CD45RO+ T helper cells fraction was negatively associated with antibody production in both groups. Conclusions: Our study provides a potential conceptual framework for monitoring vaccination efficacy in HD patients and KTRs considering the correlation established between CD19+ B cells, generation of memory CD4+ T helper cells and anti SARS-CoV2 antibody response to vaccination.

The Association of Circulating CD14++CD16+ Monocytes, Natural Killer Cells and Regulatory T Cells Subpopulations With Phenotypes of Cardiovascular Disease in a Cohort of Peritoneal Dialysis Patients.

The altered expression of immune cells including monocyte subsets, natural killer (NK) cells and CD4+CD25+ regulatory T cells (Tregs) in end-stage kidney disease, affect the modulation of inflammation and immunity with significant clinical implications. The aim of this study was to investigate the profile of specific immune cells subpopulations and their correlations with phenotypes of established cardiovascular disease (CVD), including coronary artery disease (CAD) and heart failure (HF) in peritoneal dialysis (PD) patients. Materials and Methods: 29 stable PD patients and 13 healthy volunteers were enrolled. Demographic, laboratory, bioimpedance measurements, lung ultrasound and echocardiography data were collected. The peripheral blood immune cell subsets analysis was performed using flow cytometry. Results: PD patients compared to normal controls had lower total lymphocytes (22.3 ± 6.28 vs. 31.3 ± 5.54%, p = <0.001) and B-lymphocytes (6.39 ± 3.75 vs. 9.72 ± 3.63%, p = 0.01) as well as higher CD14++CD16+ monocytes numbers (9.28 ± 6.36 vs. 4.75 ± 2.75%, p = 0.0002). PD patients with prevalent CAD had NK cells levels elevated above median values (85.7 vs. 40.9%, p = 0.04) and lower B cells counts (3.85 ± 2.46 vs. 7.2 ± 3.77%, p = 0.03). Patients with increased NK cells (>15.4%) had 3.8 times higher risk of CAD comparing with patients with lower NK cell levels (95% CI, 1.86 - 77.87; p = 0.034). B cells were inversely associated with the presence of CAD (increase of B-lymphocyte by 1% was associated with 30% less risk for presence of CAD (95% CI, -0.71 - 0.01; p = 0.05). Overhydrated patients had lower lymphocytes counts (18.3 ± 4.29% vs. 24.7 ± 6.18%, p = 0.006) and increased NK cells [20.5% (14.3, 23.6) vs. 13.21% (6.23, 19.2), p = 0.04)]. In multiple logistic regression analysis the CRP (OR 1.43; 95% CI, 1.00 - 2.05; p = 0.04)] and lymphocytes counts (OR 0.79; 95% CI, 0.63-0.99; p = 0.04)] were associated with the presence of lung comets. Patients with higher NK cells (>15.4%, n = 15) were more likely to be rapid transporters (D/P creatinine 0.76 ± 0.1 vs. 0.69 ± 0.08, p = 0.04). Patients displaying higher Tregs (>1.79%) were older (70.8 ± 10.7 years vs. 57.7 ± 14.7years, p = 0.011) and had higher nPCR (0.83 ± 0.14 vs. 0.91 ± 0.17, p = 0.09). Conclusion: Future research is required to evaluate the role of immune cells subsets as potential tools to identify patients at the highest risk for complications and guide interventions.

Hypothermic Machine Perfusion of Kidney Transplant: A Mini-Review.

Most kidney grafts are marginal and from deceased donors, which yield worse clinical outcomes. Hypothermic machine perfusion has created a paradigm shift in kidney preservation. This mini-review summarizes the main points of hypothermic machine perfusion of kidney transplants that should be known by any physician and surgeon involved with kidney transplantation. Specifically, this review explains a proposed mechanism of action of hypothermic machine perfusion of kidney transplants. This review also describes the clinical effectiveness of hypothermic machine perfusion and explains how to evaluate and predict graft functionality according to machine parameters and perfusate biomarkers. Finally, treatment options and the most recent studies on oxygenated hypothermic machine perfusion are mentioned.

Thyroid Gland as a Target of Secondary Malignancies - an Autopsy Study and Review Data.

AIM: Secondary malignancies of the thyroid gland are rarely diagnosed but their incidence at autopsy is not uncommon. MATERIALS AND METHODS: To investigate the clinicopathological features of patients with metastatic tumours of the thyroid gland, we reviewed autopsy records and pathological features of 36 cases with thyroidal secondary tumours from 266 cases of malignant neoplasias (excluding cases of primary thyroid cancer), over a 16-year period. RESULTS: There were 19 men and 17 women in the study, ranging in age from 37 to 95 years (mean 70.4 years). The incidence of metastasis in thyroid gland was 0.9% in all autopsy cases, and 13.53% of the malignant tumours. The majority were carcinomas of epithelial origin. The lung was the most common primary tumour site (33.3%), followed by the breast (8.33%) and the kidney (8.33%). The most common non-epithelial malignancy was lymphoma, followed by leukaemia (total of both 25%). As for the microscopic morphological observations, diffuse infiltration pattern of tumour cells was noted in 63.89% of the cases, the formation of nodules in 33.33% of the cases and contiguous invasion in 2.79% of the cases. There were 35.71% cases of metastases associated with multinodular goitre and 28.57% cases associated with papillary microcarcinoma. CONCLUSION: Our study indicates that thyroid secondary malignancies are not infrequent and may constitute a diagnostic problem. Lung cancer is the most common neoplasm that metastasizes to the thyroid gland in north-western Greek population.

Assessing The Predictive Value Of Regulatory Molecules For Patient Outcome In Pancreatic Cancer: A Computational Approach.

Pancreatic Cancer (PC) can be characterized as one of the most lethal cancers considering its poor diagnosis and symptoms in early stages. To assess the predictive value of regulatory molecules in terms of differentially expressed genes, we first performed a thorough search of gene expression profiling studies in pancreatic cohorts. We obtained the genes that have been identified and validated experimentally to be associated with patient outcome and also differentially expressed in tumors compared with adjacent non-tumor tissues. A two-step upstream analysis on the derived set of the genes under study was performed. The subsequent promoter and pathway analysis unveiled candidate transcription factors and regulatory molecules that potentially have regulated the detected differentially expressed genes. Predictive analysis was applied in the identified regulators and classification algorithms were implemented to model accurately patient outcome. In view of our findings, Gaussian Naïve Bayes model exhibited the highest classification accuracy and f-score concerning the predictive value of regulatory molecules in PC (accuracy =0.85, f-score =0.84).

Oxidative Stress and Acute Kidney Injury in Critical Illness: Pathophysiologic Mechanisms-Biomarkers-Interventions, and Future Perspectives.

Acute kidney injury (AKI) is a multifactorial entity that occurs in a variety of clinical settings. Although AKI is not a usual reason for intensive care unit (ICU) admission, it often complicates critically ill patients' clinical course requiring renal replacement therapy progressing sometimes to end-stage renal disease and increasing mortality. The causes of AKI in the group of ICU patients are further complicated from damaged metabolic state, systemic inflammation, sepsis, and hemodynamic dysregulations, leading to an imbalance that generates oxidative stress response. Abundant experimental and to a less extent clinical data support the important role of oxidative stress-related mechanisms in the injury phase of AKI. The purpose of this article is to present the main pathophysiologic mechanisms of AKI in ICU patients focusing on the different aspects of oxidative stress generation, the available evidence of interventional measures for AKI prevention, biomarkers used in a clinical setting, and future perspectives in oxidative stress regulation.

Cardiovascular disease: Risk factors and applicability of a risk model in a Greek cohort of renal transplant recipients.

AIM: To investigate the incidence and the determinants of cardiovascular morbidity in Greek renal transplant recipients (RTRs) expressed as major advance cardiac event (MACE) rate. METHODS: Two hundred and forty-two adult patients with a functioning graft for at least three months and available data that were followed up on the August 31, 2015 at two transplant centers of Western Greece were included in this study. Baseline recipients' data elements included demographics, clinical characteristics, history of comorbid conditions and laboratory parameters. Follow-up data regarding MACE occurrence were collected retrospectively from the patients' records and MACE risk score was calculated for each patient. RESULTS: The mean age was 53 years (63.6% males) and 47 patients (19.4%) had a pre-existing cardiovascular disease (CVD) before transplantation. The mean estimated glomerular filtration rate was 52 ± 17 mL/min per 1.73 m2. During follow-up 36 patients (14.9%) suffered a MACE with a median time to MACE 5 years (interquartile range: 2.2-10 years). Recipients with a MACE compared to recipients without a MACE had a significantly higher mean age (59 years vs 52 years, P < 0.001) and a higher prevalence of pre-existing CVD (44.4% vs 15%, P < 0.001). The 7-year predicted mean risk for MACE was 14.6% ± 12.5% overall. In RTRs who experienced a MACE, the predicted risk was 22.3% ± 17.1% and was significantly higher than in RTRs without an event 13.3% ± 11.1% (P = 0.003). The discrimination ability of the model in the Greek database of RTRs was good with an area under the receiver operating characteristics curve of 0.68 (95%CI: 0.58-0.78). CONCLUSION: In this Greek cohort of RTRs, MACE occurred in 14.9% of the patients, pre-existing CVD was the main risk factor, while MACE risk model was proved a dependable utility in predicting CVD post RT.

Aortobipopliteal bypass grafting for in situ replacement of infected aortobifemoral prosthesis.

Conventional treatment of an infected aortobifemoral graft includes total graft excision and ex situ bypass grafting, but has been associated with significant perioperative morbidity and mortality. Additionally, the presence of infection in the groin makes limb revascularization problematic. There is increasing evidence that in situ replacement of an infected graft can achieve promising results in selected patients. We present a case of an aortobifemoral graft infection, affecting both the groin as well as the entire pelvis. The patient underwent successful in situ graft replacement with a new aortobipopliteal prosthesis via an alternative extraperitoneal route.