Thursday, August 19, 2021

Ouabain Induces DNA Damage in Human Osteosarcoma U-2 OS Cells and Alters the Expression of DNA Damage and DNA Repair-associated Proteins

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In Vivo. 2021 Sep-Oct;35(5):2687-2696. doi: 10.21873/invivo.12552.

ABSTRACT

BACKGROUND/AIM: Ouabain, isolated from natural plants, exhibits anticancer activities; however, no report has presented its mechanism of DNA damage induction in human osteosarcoma cancer cells in vitro. The aim of this study was to investigate whether ouabain induces DNA damage and repair, accompanied with molecular pathways in human osteosarcoma cancer U-2 OS cells in vitro.

MATERIALS AND METHODS: The percentage of viable cell number was measured by flow cytometric assay; DNA damage was assayed by DAPI staining, comet assay, and agarose gel electrophoresis. DNA damage and repair associated protein expressions were assayed by western blotting assays.

RESULTS: Ouabain reduced total cell viability, induced chromatin condensation, DNA fragmentation, and DNA damage in U-2 OS cells. Ouabain increased p-ATMSer1981, p-ATRSer428, and p 53 at 2.5-10 μM, increased p-p53Ser15 at 10 μM; however, it decreased p-MDM2Ser166 at 2.5-10 μM. Ouabain increased p-H2A.XSer139, MDC-1, and PARP at 2.5-10 μM and BRCA1 at 5-10 μM; however, it decreased DNA-PK and MGMT at 2.5-10 μM in U-2 OS cells at 48 h treatment. Ouabain promoted expression and nuclear translocation of p-H2A.XSer139 in U-2 OS cells and this was confirmed by confocal laser microscopy.

CONCLUSION: Ouabain reduced total viable cell number through triggering DNA damage and altering the protein expression of DNA damage and repair system in U-2 OS cells in vitro.

PMID:34410957 | DOI:10.21873/invivo.12552

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Albumin-to-Alkaline Phosphatase Ratio as a Novel Prognostic Marker of Nivolumab Monotherapy for Previously Treated Metastatic Renal Cell Carcinoma

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In Vivo. 2021 Sep-Oct;35(5):2855-2862. doi: 10.21873/invivo.12573.

ABSTRACT

BACKGROUND/AIM: The relationship between albumin-to-alkaline phosphatase ratio (AAPR) and the outcome of patients with metastatic renal cell carcinoma (mRCC) treated with immune checkpoint inhibitors remains unresolved. We aimed to clarify the prognostic role of AAPR in nivolumab monotherapy for previously treated mRCC.

PATIENTS AND METHODS: We retrospectively evaluated 60 patients with mRCC treated with nivolumab after failure of at least one molecular targeted therapy. The patients were stratified into two groups based on the baseline AAPR. The threshold of AAPR was determined using receiver-operating characteristics and Youden index analyses. Overall survival (OS), progression-free survival (PFS), and objective response rate (ORR) of nivolumab therapy were compared between the high and low AAPR groups.

RESULTS: The threshold of AAPR was set at 0. 3, and 20 patients (33%) were assigned to the low AAPR group. The median OS and PFS were significantly lower in the low AAPR group than those in the high group (OS: 8.3 months vs. not reached, p<0.0001; PFS: 2.9 vs. 10.4 months, p=0.0006). Moreover, ORR was significantly lower in the low AAPR group than in the high group (16% vs. 45%, p=0.0397). Multivariate analyses further showed that AAPR was an independent factor for OS [HR=0.27 (95% CI=0.09-0.77), p=0.0151] but not for PFS (p=0.174).

CONCLUSION: Baseline AAPR was significantly associated with outcome in patients with mRCC receiving nivolumab monotherapy and may, therefore, constitute an effective prognostic factor for nivolumab treatment.

PMID:34410978 | DOI:10.21873/invivo.12573

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Establishment of an Experimental System for Intraperitoneal Chemotherapy in a Rat Model

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In Vivo. 2021 Sep-Oct;35(5):2703-2710. doi: 10.21873/invivo.12554.

ABSTRACT

AIM: To establish an experimental system for comparing different methods of intraperitoneal chemotherapy in a rat model.

MATERIALS AND METHODS: We used six-week-old Sprague-Dawley rats, and created an early postoperative intraperitoneal chemotherapy (EPIC) system using 18-gauge syringes and evacuators, and a hyperthermic intraperitoneal chemotherapy (HIPEC) system using two peristaltic pumps which controlled the flow rate and temperature. Pressurized intraperitoneal aerosol chemotherapy (PIPAC) was achieved using a nozzle for dispersing aerosols at a flow rate up to 41.5 ml/min. The distribution and intensity of 0.2% trypan blue dye was compared among three methods.

RESULTS: The distribution was limited and the intensity was weak after EPIC, and the dye stained moderately in gravity-dependent regions after HIPEC. On the other hand, the distribution was the most comprehensive, and the intensity was the greatest after PIPAC.

CONCLUSION: This experimental system in a rat model may reflect the comparative effect among EPIC, HIPEC and PIPAC in humans.

PMID:34410959 | DOI:10.21873/invivo.12554

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Pretreatment Levels of Chromogranin A and Neuron-specific Enolase in Patients With Gastroenteropancreatic Neuroendocrine Neoplasia

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In Vivo. 2021 Sep-Oct;35(5):2863-2868. doi: 10.21873/invivo.12574.

ABSTRACT

BACKGROUND/AIM: Chromogranin A (CgA) and neuron-specific enolase (NSE) are applied in the diagnosis of neuroendocrine neoplasms (NENs), especially non-functional ones. The aim of this study was to investigate the predictive values of CgA and NSE in long-term survival.

PATIENTS AND METHODS: Our retrospective analysis included 65 patients with histologically verified gastroenteropancreatic NEN between 2005 and 2019. We performed bivariate and multivariable analyses to evaluate the relationship between CgA and NSE values before histological assessment and overall survival. Distribution of time-to-event was analyzed using Kaplan-Meier survival curves and modelled by Cox regression models.

RESULTS: Elevated NSE levels prior to histology were significantly associated with worse survival (HR=1.13, p=0.004) and were associated with low-differentiated NENs ( rs=0.321, p=0.0338). CgA was associated with well-differentiated tumors (rs=0.233), but not significantly.

CONCLUSION: Pretreatment serum levels of NSE can serve as a valuable additional predictor of long-term survival in patients with NEN.

PMID:34410979 | DOI:10.21873/invivo.12574

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In Vivo Comparison of Three Human Acellular Dermal Matrices for Breast Reconstruction

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In Vivo. 2021 Sep-Oct;35(5):2719-2728. doi: 10.21873/invivo.12556.

ABSTRACT

BACKGROUND/AIM: Acellular dermal matrices (ADMs) have become popular in implant-based breast reconstruction. The aim of this study was to compare three commonly used ADM products in vivo in an animal model.

MATERIALS AND METHODS: The nucleic acid content (residual double-stranded DNA) and the levels of the remaining growth factors after decellularization were measured for each ADM. Cytocompatibility with ADMs was documented using NIH 3T3 mouse fibroblast cells. In vivo, the implanted ADMs were histologically evaluated at 1, 2, 3, and 6 months (n=5) using male 8-week-old Sprague-Dawley rats.

RESULTS: Fibroblasts grew in the SureDerm HD and DermACELL with no cytotoxicity. In a rat model, SureDerm HD and DermACELL incorporated more readily into the surrounding host tissue, as measured by rapid cell influx and collagen deposition, and showed more delaye d tissue remodeling with decreased matrix metalloproteinases levels compared to AlloDerm.

CONCLUSION: SureDerm HD and DermACELL can be used as biological materials for breast reconstruction.

PMID:34410961 | DOI:10.21873/invivo.12556

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Clinical Outcomes of Mixed Response to Pembrolizumab in Advanced Urothelial Carcinoma After Platinum-based Chemotherapy

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In Vivo. 2021 Sep-Oct;35(5):2869-2874. doi: 10.21873/invivo.12575.

ABSTRACT

BACKGROUND/AIM: Despite the presence of a mixed response (MR) in patients with urothelial carcinoma (UC) who receive immune checkpoint inhibitors, the clinical outcome of these patient has not been reported. We evaluated the clinical outcome of MR to pembrolizumab for advanced UC.

PATIENTS AND METHODS: Advanced UC patients who received pembrolizumab after platinum-based chemotherapy failure with measurable disease in multiple organs were retrospectively analyzed.

RESULTS: Among 31 patients, MR [including progressive disease (PD)+complete response (CR) or partial response (PR)] was confirmed in 4 (12.9%). The median overall survival (OS) of the CR+PR (including CR+SD±PR), stable disease (SD), PD (including PD±SD) and MR groups was 16.0, 5.1, 5.4 and 4.3 months, respectively. There was no significant difference in the OS between the MR and CR+PR res ponse groups (log-rank test, p=0.069).

CONCLUSION: A mixed response to pembrolizumab in advanced UC was not uncommon. Despite the non-significant difference in the OS between the mixed and CR+PR response groups, the OS of the MR group tended to be similar to that of the SD and PD response groups.

PMID:34410980 | DOI:10.21873/invivo.12575

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Postoperative Complications in Breast Reconstruction With Porcine Acellular Dermis and Polypropylene Meshes in Subpectoral Implant Placement

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In Vivo. 2021 Sep-Oct;35(5):2739-2746. doi: 10.21873/invivo.12558.

ABSTRACT

AIM: This research compares postoperative complication rates with Strattice™, SERAGYN® BR, and TiLOOP® Bra interposition devices for subpectoral implant placement after skin or nipple sparing mastectomy.

PATIENTS AND METHODS: 188 breast reconstructions in 157 patients after primary (n=96), secondary (n=71), or prophylactic (n=21) surgery were analyzed regarding major and minor complications.

RESULTS: With acellular dermal matrix (ADM) Strattice™, 27.5% major and 27.5% minor complications occurred. Implant loss rates were 27.3% in primary and 30.8% in secondary reconstructions. With SERAGYN® BR, 11.1% major and 13,0% minor complications occurred. Implant losses (6.1%) occurred exclusively in primary reconstructions. With TiLOOP® Bra, 14.9% major and 9.6% minor complications occurred. Implant loss ra tes were 7.7% in primary and 7.1% in secondary reconstructions.

CONCLUSION: ADM was associated with high complication rates in primary and secondary reconstructions. Low complication rates were seen with mesh interposition devices in primary, secondary, and prophylactic reconstructions.

PMID:34410963 | DOI:10.21873/invivo.12558

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Ten Daily Fractions for Whole Breast Cancer Irradiation: Long Term Results

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In Vivo. 2021 Sep-Oct;35(5):2875-2880. doi: 10.21873/invivo.12576.

ABSTRACT

BACKGROUND/AIM: To report the feasibility and oncological outcomes in breast cancer patients treated with a short hypofractionated radiotherapy schedule.

PATIENTS AND METHODS: We evaluated 380 breast cancer patients treated with ten daily fractions of radiotherapy up to 39 Gy on tumor bed. Primary endpoint was local relapse rate (LRR). Secondary endpoints were overall survival (OS) and metastasis-free survival (MFS).

RESULTS: The median follow up was 5.0 years. Two- and 5-year LRR rates were 0.2 and 2%, respectively. Two- and 5-year MFS rates were 96.1% and 90.5%, respectively. Two and 5-year OS rates were 97.4% and 95%, respectively.

CONCLUSION: This short schedule may represent an alternative option to standard mild hypofractionated radiotherapy in breast cancer patients due to its excellent feasibility and very low recurrence rate.

PM ID:34410981 | DOI:10.21873/invivo.12576

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Neutrophil Gelatinase-associated Lipocalin Predicts Post-traumatic Acute Kidney Injury in Severely Injured Patients

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In Vivo. 2021 Sep-Oct;35(5):2755-2762. doi: 10.21873/invivo.12560.

ABSTRACT

BACKGROUND: Early detection of acute kidney injury (AKI) is crucial in the management of multiple-organ dysfunction syndrome in severely injured patients. Standard laboratory parameters usually increase with temporal delay. Therefore, we evaluated neutrophil gelatinase-associated lipocalin (NGAL) as an early marker for acute kidney injury.

PATIENTS AND METHODS: We retrospectively evaluated patients admitted to a level 1 trauma center. We collected clinicodemographic data and measured kidney-related factors and plasma cytokines.

RESULTS: A total of 39 patients were included. Patients with AKI had significantly higher levels not only of serum creatinine and urea, but also of NGAL (all p<0.001) than patients without AKI. The optimal NGAL cut-off value was determined to be 177 ng/ml, showing significant correlation with imminent or manifest AKI (p< ;0.001). Other independent markers correlated with AKI included pre-existing chronic kidney disease, use of catecholamines, and severe injury (p<0.001).

CONCLUSION: The serum level of NGAL is feasible early predictor of AKI.

PMID:34410965 | DOI:10.21873/invivo.12560

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A New and Validated, Randomised, Controlled Placebo Water Development Trial for Medicinal Water-based Research

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In Vivo. 2021 Sep-Oct;35(5):2881-2888. doi: 10.21873/invivo.12577.

ABSTRACT

BACKGROUND/AIM: To develop and validate an easy-to-use and cheap method capable of producing placebo from tap water for medicinal water efficacy trials.

PATIENTS AND METHODS: Patients were divided into two groups, medicinal water and tap water group. A single 20-minute-long treatment was performed in bathtubs. Patients were asked four times during the bath to tell if they were treated with medicinal water, tap water, or could not decide. Patients were scored, one point was given for each correct answer.

RESULTS: A total of 174 patients were enrolled. No significant differences were found either between the average scores or the answers of the two groups. Being familiar with the Harkány medicinal water did not influence the rate of correct answers either. There was no statistically significant difference in the number of changes of opinions between the two groups.

CONCLUSION: The used method is appropriate for producing a validated placebo from tap water.

PMID:34410982 | DOI:10.21873/invivo.12577

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Clinical Significance of TAP1 and DLL4 Expression in Patients With Locally Advanced Gastric Cancer

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In Vivo. 2021 Sep-Oct;35(5):2771-2777. doi: 10.21873/invivo.12562.

ABSTRACT

BACKGROUND/AIM: Cancer stem cells (CSCs) are reported to associated with cancer metastasis, relapse, and chemoresistance. This study examined the clinical significance of the expression of two CSC markers, the transporter associated with antigen processing 1 (TAP1) and the Delta-like 4 (DLL4) protein, in patients with locally advanced GC.

PATIENTS AND METHODS: This study was performed using samples obtained from 413 pathological stage II/III GC patients after curative gastrectomy. We examined TAP1 and DLL4 expression using immunohistochemical analysis with tissue microarray and examined the association between TAP1 or DLL4 expression, clinicopathological factors and survival.

RESULTS: High TAP1 expression was associated with better overall survival compared to low TAP1 expression (p=0.004). Furthermore, in multivariate analysis, high TAP1 expression was defined as a predictive factor for good survival. There was no significant difference between DLL4 expression and clinicopathological features and overall survival.

CONCLUSION: TAP1 expression may be a useful prognostic marker in patients with locally advanced GC.

PMID:34410967 | DOI:10.21873/invivo.12562

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