The results of designing a new prototype device and algorithm in closed method intraperitoneal hyperthermia model in rats.

BACKGROUND: There is currently no standard medical device and method available for hyperthermic intraperitoneal therapy studies in rats. In this study, we present our designed device and algorithm that operates based on our own protocol for hyperthermic intraperitoneal treatment in rats. The aim was to demonstrate the effectiveness of the designed device, algorithm, and hyperthermia protocol by showing that the device can achieve the desired temperature inside the rat's abdomen, does not cause rat loss due to complications, operates autonomously, and provides warnings to the operator in case of emergencies. METHODS: A closed method for intraperitoneal hyperthermia protocol was established for 6 female 8-week-old (280-310 g) albino Wistar rats. Fluid inlet and outlet tubes and a temperature probe were inserted through a 1 cm vertical incision between the xiphoid and bladder in the rat's abdomen, and the skin was sutured in a circular manner. A protocol for intraperitoneal hyperthermic treat-ment was established using a saline solution at a flow rate of 100 mL/min for 60 min, maintaining a temperature of 41°C±0.5 inside the rat's abdomen. RESULTS: During the study, a temperature of 41°C±0.5 was successfully achieved in the abdomen of all rats at a flow rate of 100 mL/min±5 for 60 min. Due to three rats reaching a rectal temperature above 38.5°C during the hyperthermia protocol, external cooling was applied to the rat's tail base using ice. There were no losses until the postoperative 72nd h, and the study was successfully completed. CONCLUSION: Our designed device and algorithm, which prioritize animal welfare, operate rapidly, safely, and with high accuracy sensitivity, have been successful in hyperthermic intraperitoneal treatment studies in rats. We believe that they can be used as a stan-dard method and approach in hyperthermic intraperitoneal studies in rats.

Evaluation of muscle mass in obesity, prediabetes and diabetes mellitus by different equations used for the measurement of muscle mass.

OBJECTIVE: Insulin resistance is one of risk factors for sarcopenia and there is no specific equation for the measurement of muscle mass. The present study aimed to evaluate muscle mass in the patients with obesity, prediabetes (PDM) and type 2 diabetes mellitus (DM) by different equations for the measurement of muscle mass. METHODS: Obese patients aged 18-65 years old, who presented between 2013 and 2015 were reviewed and they were separated into three groups as obese, prediabetes (PDM) and diabetes mellitus (DM). Height, body weight, body mass index (BMI), sum of the appendicular lean masses (ALM) were measured in all participants. Body muscle mass ratio was calculated as the total muscle mass divided by the body weight, and skeletal muscle index was calculated as the total muscle mass divided by the square of the height. In addition, ALM/weight, ALM/height2 and ALM/BMI ratios were also evaluated. RESULTS: A total of 1107 participants, of whom 666 (60.2%) were female, were enrolled into the study. Of the participants, 288 (%26.02) had obesity, 524 (%47.33) had PDM and 295 (26.65%) had DM. There was a significant difference in ALM/BMI ratio between the three groups for both genders (p = 0.003 for female and p = 0.003 for male). ALM/weight ratio and body muscle mass ratio were decreased between groups in female, whereas it was no difference in male (p = 0.003, p < 0.001 for females, respectively; p = 0.802, p = 0.840 for males, respectively). CONCLUSIONS: ALM/BMI may be more accurate for the evaluation of muscle mass in middle-aged obese, PDM and DM subjects.

6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase-3 is required for transforming growth factor ß1-enhanced invasion of Panc1 cells in vitro.

Transforming growth factor ß1 (TGFß1) is a well-established inducer of the epithelial-mesenchymal transition (EMT) that is essential for the acquisition of malignant properties, such as invasion, in tumor cells. Although recent studies suggest that the EMT in tumor cells is associated with reprogramming of energy metabolism and TGFß1 has been shown to stimulate glycolysis in multiple primary cell lines, little is known about TGFß1's effect on glycolysis and glycolytic regulators in transformed cells. Given the known regulatory role of 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase-3 (PFKFB3) in glycolysis and association of glycolytic activity with malignant features such as invasion, we sought to investigate whether TGFß1 regulates PFKFB3 expression and if PFKFB3 is involved in the TGFß1-mediated increase in the invasive ability of the Panc1 cell cline-a well-established model of TGFß1-initiated EMT. Herein we demonstrate that TGFß1 induces PFKFB3 expression and stimulates glycolysis in Panc1 cells. We also show that siRNA silencing of PFKFB3 prevents the stimulation of glycolysis and in vitro invasive ability of Panc1 cells by TGFß1. Furthermore, PFKFB3 silencing suppresses the TGFß1-mediated induction of the Snail protein, suggesting that PFKFB3 is required for the regulation of Snail expression by TGFß1. Taken together, our study identifies PFKFB3 as a key TGFß1 effector protein that mediates TGFß1's effect on Snail expression, invasion, and glycolysis.

Impact of 3'UTR variation patterns of the KRAS gene on the aggressiveness of pancreatobiliary tumors with the KRAS G13D mutation in a Turkish population.

OBJECTIVE: Several studies have demonstrated the importance of mutations in codons 12, 13 and 61 and variations in the 3' untranslated region (3'UTR) of the KRAS gene, frequently observed genetic events in the progression of pancreatobiliary tumors (PBT). However, limited data exist on the clinical effect of these alterations. The aim of the current study was to clarify the frequency of relevant alterations of the 3'UTR regions of the KRAS gene and the effect of KRAS 3'UTR polymorphisms on the prognosis of patients with codon 12, 13 and 61 mutations in a Turkish population with PBT. METHODS: Codons 12, 13, and 61 and 3'UTRs of the KRAS gene were screened by single-strand conformation polymorphism (SSCP) analysis and DNA sequencing in 43 patients and 10 controls. Chi-squared and independent sample T tests were used to evaluate the results of the mutation analysis and clinical features of the patients. RESULTS: We defined the c.38G > A (rs112445441, p.G13D) (39.54%) mutation and two 3'UTR variations, c.*4066delA (rs560890523) (23.26%) and c.*4065_*4066delAA (rs57698689) (6.98%), in the KRAS gene of Turkish patients. There was a statistically significant relationship between the c.*4066delA (rs560890523) and c.*4065_*4066delAA (rs57698689) variations and invasion and lymph node metastasis status of the patients (p < 0.001). Compared to patients with c.38G > A (rs112445441, p.G13D), patients with c.*4066delA (rs560890523) and c.38G > A (rs112445441, p.G13D) presented more aggressive tumors with highly invasive features. The present study contributes findings regarding the clinical effects of KRAS alterations in PBT. Based on our study, further investigations are required.

Expression and clinical significance of miRNAs that may be associated with the FHIT gene in breast cancer.

The dysregulation of miRNA expression has frequently been observed in breast cancer. Therefore, we investigated the expression profile of miRNAs that may be associated with expression of the FHIT gene in breast cancer and assessed their clinicopathological significance. The expression levels of miR-143, miR-663a, miR-668, miR-922 and FHIT were analyzed in normal and malignant breast tissues from 65 patients with breast cancer. We studied the correlation between the expression of miR-143, miR-663a, miR-668, miR-922 and FHIT and the clinicopathological features presented by the patients. The expression levels of the miRNAs and FHIT were downregulated in breast cancer tissue. The expression levels of miR-143, miR-663a and miR-668 were significantly reduced in FHIT downregulated tumors. miR-668 expression was also significantly altered relative to FHIT down- and up- regulated tumor tissues. Reduced miR-663a expression was statistically associated with high-grade ER/PR (+) status, benign reactive hyperplasia, lymph-node metastasis, in-situ component >25% and Ki 67>15% compared with non-tumor tissues. Additionally, reduced miR-668 expression was significantly different between tumors with and without lymph-node metastasis. miR-668 may play an important role in breast cancer development and progression by regulating the expression of FHIT. Furthermore, miR-668 and miR-663a may be potential prognostic biomarkers for breast cancer.

miR-216b Targets FGFR1 and Confers Sensitivity to Radiotherapy in Pancreatic Ductal Adenocarcinoma Patients Without EGFR or KRAS Mutation.

OBJECTIVES: The success of gemcitabine plus radiotherapy is dependent on the mutation status of pancreatic ductal adenocarcinoma (PDAC) tumors in the EGFR and KRAS genes; however, radiotherapy resistance may also be modulated epigenetically by microRNA (miRNA) regulation. In this study, we examined the potential effect of miRNAs on the resistance to radiotherapy in cases without EGFR or KRAS mutation. METHODS: The association of EGFR and KRAS mutation status and different expression patterns of 6 selected miRNAs related to the EGFR/KRAS signaling pathway were evaluated in the tumors of 42 patients with PDAC. RESULTS: Reduced miR-216b and miR-217 expression was associated with aggressive tumor characteristics and shortened disease-free survival. In addition, miR-216b expression was reduced 2.7-fold in the cases that did not benefit from therapy, although they did not demonstrate EGFR or KRAS expression (P = 0.0316). A negative correlation between FGFR1 and miR-216b expression (r = -0.355) was found in the tumors of these cases. CONCLUSIONS: Further studies and validations are required; in the tumors of patients with PDAC without activating mutations and induced expression of EGFR/KRAS genes, down-regulated miR-216b expression may be associated with a poor response to radiotherapy via deregulation of another signaling pathway related to FGFR1 signaling.

Association of miR-1266 with recurrence/metastasis potential in estrogen receptor positive breast cancer patients.

The Homeobox B13 (HOXB13):Interleukin 17 Receptor B (IL17BR) index of estrogen receptor (ER)-positive breast cancer (ER (+) BC) patients may be a potential biomarker of recurrence/ metastasis. However, effects of microRNA (miRNA) binding to the 3' untranslated region (3 ?UTR) of HOXB13 and IL17BR and its function on recurrence/metastasis in ER (+) BC remains elusive. The aims of this study were to determine the expression of miRNAs that bind to 3 ?UTR of HOXB13 and IL17BR in ER (+) BC patients and asess the effects of these miRNAs on recurrence/metastasis. The expression profiles of HOXB13 and IL17BR were evaluated using RT- PCR in tumors and normal tissue samples from 40 ER (+) BC patients. The expression level of 4 miRNAs, which were predicted to bind the 3 ?UTR of HOXB13 and IL17BR using TargetScan, microRNA.org and miRDB online databases, were further evaluated with RT-PCR. Our findings demonstrated that high miR-1266 levels might be significant prognostic factor for recurrence/metastasis occurrence (3.05 fold p=0.004) and tamoxifen response (3.90 fold; p=0.2514) in ER (+) BC cases. Although we suggest that modulation of miR-1266 expression may be an important mechanism underlying the chemoresistance of ER (+) BC, advanced studies and validation are required.