Adam Rzechonek

Cultivation of circulating tumor cells in esophageal cancer

The presence of circulating tumor cells (CTCs) in patients with metastatic carcinoma is generally associated with poor clinical outcome. There have been many investigations showing a possible use of CTCs as minimally invasive predictive and prognostic biomarker in cancer medicine. In this report a size-based method (MetaCell®) for quick and easy enrichment and cultivation of CTCs is presented to enable possible CTCs use in esophageal cancer (EC) management. In total, 43 patients with diagnosed EC, 20 with adenocarcinoma (AdenoCa) and 23 with squamous cell carcinoma (SCC), were enrolled into the adaptive prospective-like study .All the patients were candidates for surgery. The CTCs were detected in 27 patients (62.8%), with a higher rate in adenocarcinoma (75%) than SCC (52%). Finally, there were 26 patients with resectable tumors exhibiting CTCs-positivity in 69.2% and 17 patients with non-resectable tumors with 41.7% CTCs-positivity. Interestingly, in the patients undergoing neoadjuvant therapy, the CTCs were detected at time of surgery in 55.5% (10/18). The overall size-based filtration approach enabled to isolate viable CTCs and evaluate to their cytomorphological features by means of vital fluorescent staining. The CTCs were cultured in vitro for further downstream applications including immunohistochemical analysis. This is the first report of the successful culturing of esophageal cancer CTCs. The detection of CTCs presence could help in the future to guide timing of surgical treatment in EC patients.

Retrosternal goiter located in the mediastinum: surgical approach and operative difficulties

Most retrosternal goiters are situated in the anterior mediastinal compartment, but according to the literature, 10-15% are located in the posterior mediastinum. Although most of the anterior mediastinal goiters can be removed by a transcervical approach, posterior mediastinal goiters may require additional extracervical incisions. We report the case of a huge posterior mediastinal goiter extending from the neck retrotracheally beyond the aortic arch and azygous vein with crossover from the left to the right side and ending at the level of the lower part of the left cardiac atrium, nearly reaching the diaphragm. Surgical removal is the treatment of choice in such cases. We performed an operation using a transcervical and right thoracotomy approach. Histopathological examination confirmed the diagnosis of the large goiter. The patient recovered well and was discharged in 1 week.

Cell cycle-dependent expression and subcellular localization of fructose 1,6-bisphosphatase

Recently a gluconeogenic enzyme was discovered—fructose 1,6-bisphosphatase (FBPase)—that localizes in the nucleus of a proliferating cell, but its physiological role in this compartment remains unclear. Here, we demonstrate the link between nuclear localization of FBPase and the cell cycle progression. Results of our studies indicate that in human and mouse squamous cell lung cancer, as well as in the HL-1 cardiomyocytes, FBPase nuclear localization correlates with nuclear localization of S and G2 phase cyclins. Additionally, activity and expression of the enzyme depends on cell cycle stages. Identification of FBPase interacting partners with mass spectrometry reveals a set of nuclear proteins involved in cell cycle regulation, mRNA processing and in stabilization of genomic DNA structure. To our knowledge, this is the first experimental evidence that muscle FBPase is involved in cell cycle events.

Limited Clinical Significance of Dimeric Form of Pyruvate Kinase as a Diagnostic and Prognostic Biomarker in Non-small Cell Lung Cancer

Metabolism of tumor tissue differs from the normal one by the intensity of protein synthesis and glycolysis. The dimeric pyruvate kinase (PKM2) is a specific enzyme for tumor glycolysis. The aim of this study was to determine the relationship between the activity of PKM2 and the type and stage of non-small cell lung cancer (NSCLC). A second objective was to compare the expression of PKM2 with disease progression and prognosis. We studied 65 patients divided into two groups: 45 patients with lung cancer and 20 non-cancer healthy subjects taken as control. The serum activity of PKM2 was assessed spectrophotometrically. We found that PKM2 activity was greater, on average, by 136 % for adenocarcinoma and for 126 % for squamous cell carcinoma compared with that present in control subjects. The higher PKM2 activity was associated only with Stage III of cancer (p < 0.001). Sensitivity of PKM2 as a cancer marker was 79 % for adenocarcinoma and 81 % for squamous cell carcinoma and specificity was 50 % for both cancer types. We conclude that PKM2 activity is higher in patients with NSCLC than in healthy subjects. The level of PKM2 activity is associated with advanced stage of cancer. Nonetheless, low specificity of PKM2 assessment makes it of limited utility in NSCLC diagnosis or evaluation of cancer progression.

Treatment of Recurrent Primary Spontaneous Pneumothorax – Own Experience

UNLABELLED Primary spontaneous pneumothorax could be a serious therapeutic problem in case of recurrence. Lack of therapeutic standards sometimes leads to delay in definitive surgical treatment and could cause respiratory complications. The aim of the study was the evaluation of treatment results in patients with recurrence of primary spontaneous pneumothorax and looking for optimal therapeutic method after first recurrence (surgical treatment vs. pleural drainage). MATERIAL AND METHODS Between 01.01.2009 and 31.07.2010 fifty four patients with recurrent primary spontaneous pneumothorax was hospitalized in Wrocław Thoracic Surgery Centre (24.3% of all patients with pneumothorax). The recurrence was treated surgically in 24 cases, in 30 pleural drainage was performed: simple drainage (n=14) or drainage with chemical pleurodesis (n=16). Mean age of patients treated without surgery was higher than surgically treated (p=0,012). RESULTS In surgery group no recurrence was found, in drainage group 11 recurrences occurred (p=0.0009). In group of 11 patients with second recurrence, pleurodesis was performed four times (36%) vs. 12 times (63%) in 19 patients without a recurrence of the disease. 70% of non-surgically treated patients vs. 50% of surgically treated were afraid of recurrence (p=0.01). Among 11 patients in drainage group, nine underwent surgery at the second episode of recurrence. CONCLUSIONS The optimal treatment method in case of first recurrence of primary spontaneous pneumothorax is surgical treatment. When it is not possible chemical pleurodesis should be performed during pleural drainage. Most of the patients after second recurrence are treated surgically anyway. The surgical treatment significantly reduces patients fears for future recurrence of the disease. Younger patients are most often surgically treated.

Immunomagnetic detection of cancer cells in pleural effusion of generalized cancer.

Malignant pleural effusions (MPE) are a common clinical problem in patients with neoplastic disease. Pleural fluid cytology is the simplest definitive method for obtaining a diagnosis of MPE. We describe a method that may increase the cancer cell detection rate using immunomagnetic separation in MPE. In comparison to standard MPE cytodiagnostic methods, we report a more streamlined method of isolation living cells that are able to proliferate. These captured cells can then be used for additional downstream analysis e.g. chemosensitivity testing. Several case studies of MPE diagnostics using immunomagnetic separation are presented in the following report. The immunomagnetic separation of cancer cells from MPE could be used for more accurate staging of patients with routine effusions.

New therapy of pleural empyema by deoxyribonuclease

Empyema is a severe complication of different diseases and traumas. Management of this complication is difficult and should comprise general and local procedures. The general procedure is mainly based on administering wide-spectrum antibiotics. Local management depends on patient general condition, but in all cases the essential procedure is to insert a drain into the pleural cavity and to evacuate the pus. Sometimes pus is very thick and its evacuation and following re-expansion of the lung is rather impossible. In these patients surgical intervention is needed. The use of intrapleural enzymes to support the drainage was first described in 1949 by Tillett and Sherry using a mixture of streptokinase and streptococcal deoxyribonuclease. Nowadays, purified streptokinase has come into widespread use, but recent studies reported no streptokinase effect on pus viscosity. On the other side, deoxyribonuclease reduces pus viscosity and may be more useful in treatment. We report two cases of intrapleural administration of Pulmozyme (alfa dornase - deoxyribonuclease (HOFFMANN-LA ROCHE AG) in dosage 2×2.5mg with a significant improvement caused by changes in pus viscosity.

Surgical Correction of Pectus Excavatum by the Nuss Procedure: A 15-Year Experience Study

Pectus excavatum is the most common congenital deformity of the chest. The Nuss procedure is minimally invasive surgical correction of this defect, using retrosternal metal bars. The purpose of the present study was to describe a 15-year experience with the Nuss surgery, and to evaluate the long-term clinical results of the procedure. We retrospectively evaluated 239 patients, aged 14-34, who underwent the Nuss surgery in the years 2002-2016. Postoperative complications were observed in 40/236 (16.9%) patients. The most common complication was pneumothorax in 14/239 patients. Less common were the following: wound infection in 4, pleural effusion in 3, allergy to nickel in 1, lung atelectasis in 1, and ventricular failure in 1 patient. Three patients were treated because of severe postoperative pain, and in one case the implant had to be removed. Postoperative complications associated with the number of bars inserted, but not with the patient age or gender. A satisfactory and long lasting corrective effect of surgery was observed in 231/239 (96.7%) of patients. There was no perioperative mortality. We conclude that the Nuss surgery is a safe surgery that demonstrates excellent and long-lasting esthetic results, with a low risk of severe complications.

Electroporation with Cisplatin against Metastatic Pancreatic Cancer: In Vitro Study on Human Primary Cell Culture

Despite the rapid progression of cancer pharmacotherapy, the high drug resistance of pancreatic ductal adenocarcinoma (PDA) makes it one of the most lethal malignancies. Therefore, there are high expectations associated with experimental therapies, such as electrochemotherapy (ECT). This technique involves the application of short electric pulses to induce transitional permeabilization of the cellular membrane, thus enhancing drug molecules influx. The aim of the study was to investigate the influence of electroporation with cisplatin (CisEP) on the primary culture of human PDA cells from lung metastases—their survival and stress response. Considering the growing importance of various research models, two established human PDA cell lines, EPP85-181P (sensitive to daunorubicin) and EPP85-181RDB (resistant to daunorubicin), were utilized as a reference control. Cisplatin revealed higher cytotoxicity towards established cell lines. Following CisEP application, we observed a significant decrease of cells viability in the primary culture model. After CisEP therapy, an increased immunoreactivity with SOD-2 and Casp-3 antibodies was noticed. In conclusion, we discovered that electroporation can enhance the cytotoxic effect of cisplatin in pancreatic cancer cells in vitro. This effect was evident for cells from the primary culture. The obtained results confirm the importance of primary cells models in studies on the efficacy of experimental cancer therapies.

Influence of miR-7a and miR-24-3p on the SOX18 transcript in lung adenocarcinoma

The molecular pathogenesis of the development of non-small cell lung carcinomas (NSCLCs) is extremely complex. Understanding the molecular basis of the development of this malignant tumor may enable the use of targeted therapy, which may result in a better treatment outome for these patients. Adenocarcinoma (AC) is the most common NSCLC subtype, equally common among smokers and non-smokers, and its pathogenesis remains unknown. The SOX18 protein is an important protein that plays a role in the development of blood and lymphatic vessels during the process of embryogenesis. Recent studies have also shown that the SOX18 protein may play a significant role in tumors, including lung cancers. In the present study, we analyzed the expression of the SOX18 protein and the mRNA level in postoperative samples of AC and non-malignant lung tissues (NMLTs), and a disparity in both levels was observed. Based on our previous observations that miR-7a and miR-24-3p are able to modulate SOX18 expression in NSCLC, the main aim of this study was to verify the miRNA modulation of the SOX18 transcript with the use of the MirTrap System in established lung cancer cell lines NCI-H1703, NCI-H522 and A549. The SOX18 mRNA expression level was significantly lower in AC than that noted in the NMLTs (P<0.0001). However, the protein levels were higher in AC cases compared to levels noted in the NMLTs (P<0.0001). Additionally, correlations between the RQ values of SOX18 in NMLT and AC cases (r=0.8195, P=0.0001), and between miR-7a and miR24-3p in AC cases (r=0.4344, P=0.0016), were noted. In conclusion, we confirmed that miR-7a and miR-24-3p are more highly expressed in NMLTs than in the AC samples, and that they modulate the SOX18 transcript in NSCLC cells.