Dariusz Waniczek

Specific metabolic biomarkers as risk and prognostic factors in colorectal cancer.

Advances in genomics, molecular pathology and metabolism have generated many candidate biomarkers of colorectal cancer with potential clinical value. Epidemiological and biological studies suggest a role for adiposity, dyslipidaemia, hyperinsulinemia, altered glucose homeostasis, and elevated expression of insulin-like growth factor (IGF) axis members in the risk and prognosis of cancer. This review discusses some recent past and current approaches being taken by researches in obesity and metabolic disorders. The authors describe three main systems as the most studied metabolic candidates of carcinogenesis: dyslipidemias, adipokines and insulin/IGF axis. However, each of these components is unsuccessful in defining the diseases risk and progression, while their co-occurrence increases cancer incidence and mortality in both men and women.

Adjunct Methods of the Standard Diabetic Foot Ulceration Therapy

The outcome of management of diabetic foot ulceration (DFU) is poor and insufficient. DFU therapy includes the standard management as debridement of the wound, revascularization procedures, off-loading of the ulcer and antibacterial actions, and supplementation of growth factors and cytokines, leading to stimulation of granulation, epidermization, and angiogenesis. The aim of the present review is to summarize the adjunct methods of the standard DFU therapy as hyperbaric oxygen therapy (HBOT), maggot therapy (MT), and platelet-rich plasma therapy (PRPT). The results of preclinical and clinical trials indicated that the methods may reduce time of therapy, short-term morbidity, and the risk of major amputation.

Clinical Significance and Prognostic Relevance of Microsatellite Instability in Sporadic Colorectal Cancer Patients

Microsatellite instability (MSI) is a marker of the replication error phenotype. It is caused by impaired DNA mismatch repair processes (MMR), resulting in ineffectiveness of the mechanisms responsible for the DNA replication precision and postreplicative DNA repair. MSI underlies the pathogenesis of 10%–20% of colorectal cancer (CRC) cases. The data about the potential value of MMR status as a predictive factor for 5-fluorouracil (FU)-based chemotherapy remain unclear. According to National Comprehensive Cancer Network updated guidelines, MSI testing is recommended for all patients with stage II CRC because patients with MSI-H (high-frequency MSI) tumour may have a good prognosis and obtain no benefit from 5-FU-based adjuvant chemotherapy. The significance of the MSI status as a predictive factor for patients with metastatic disease was not confirmed. The association between the MSI status and the efficacy of the therapy based on anti-programmed death-1 receptor inhibitors requires further studies.

Potential role of human papilloma virus in the pathogenesis of gastric cancer

AIM To demonstrate the presence and biological activity of human papilloma virus (HPV) in gastric cancer (GAC) tissues. METHODS The study involved 84 surgically treated patients with gastric adenocarcinoma, regardless of the clinical stage of the disease. The presence of HPV DNA of high oncogenic risk types in formalin-fixed, paraffin-embedded tumor samples was determined using quantitative polymerase chain reaction analysis. A stringent protocol of prevention of cross- and environmental contamination was applied during DNA isolation, and amplification, as well as confirmation of the biological activity of the virus in tumor cells, was implemented. The study utilized the Real-time High Risk HPV test, which detects the DNA of 14 HPV subtypes that are considered to have high oncogenic potential. The overexpression of the p16(INK4a) protein assessed immunohistochemically was considered confirmation of the HPV infection. RESULTS Among the 89 patients initially included in the study group, diagnostic results were obtained for 84 individuals. In five cases, either the histopathological material was too scant to isolate the necessary amount of DNA, or the isolated DNA was significantly degraded, resulting in the failure of internal control amplification within the predefined number of 35 cycles. Those patients were excluded from further analysis. The amplification of HPV DNA was demonstrated in none of the 84 tissue samples; thus, all cases were considered to have a negative DNA status of highly oncogenic HPV subtypes. Immunohistochemical staining provided diagnostic results for all of the examined tissue samples, and excluded the accumulation of the p16(INK4a) protein in tumor cells, thus confirming the lack of active HPV infection in all of the individuals. CONCLUSION The study does not confirm the presence or biological activity of HPV in tumor tissues. Thus, the relationship between GAC and HPV infection, in the Central European population seems doubtful.

Analysis of PTEN expression in large intestine polyps and its relation to the recognized histopathological and clinical risk factors for cancer development in this location

Aim of the study PTEN is an important gene whose protein product is double specific phosphatase holding key regulatory functions in sending signals from membrane receptors for growth factors into the cell downstreams. Its participation, mainly by PI3K/AKT signaling pathway in the pathomechanism of many malignant cancers was unambiguously confirmed. The PTEN function gets disturbed on many levels and for various reasons. Disorders of PTEN protein expression seem to be even more common in many carcinomas. The aim of the study is to enquire the meaning of PTEN expression in the cancer transformation process in large intestine glandular polyps. Material and methods The group includes 40 patients, 21 men and 19 women, age median 64 years (51–83) qualified to endoscopic removal of large intestine polyp. Tissue material obtained during polyp removal endoscopy was immediately fixed in 4% buffered formalin solution with the mixture of phosphatase activity inhibitors (PhosStop Roche). Time of fixation 24–48 h. After fixation, the material was embedded in paraffin. PTEN visualization was based on specific rabbit monoclonal antibodies (Cell Signaling). The expression of PTEN protein in large intestine and rectum polyps was marked by a semi-quantitative method and an attempt to correlate the results with the acknowledged clinical and histopathological malignancy risk factors was undertaken. Results Loss or weakening of protein expression was found in 45% cases. Moreover, the relationship between polyp diameter and a loss of PTEN expression was proved. The received results can indicate a significant participation of PTEN gene in early oncogenesis stages of large intestine cancer.

Assessment of PI3K/AKT/PTEN signaling pathway activity in colorectal cancer using quantum dot‑conjugated antibodies

In certain patients with advanced colorectal cancer, loss of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) activity is observed. PTEN is a major gatekeeper gene of the AKT serine/threonine kinase (AKT) signaling pathway responsible for the proliferative activity of cells. The assessment of AKT activity may be a prognostic factor or a predictor of response to the targeted therapies against particular signaling proteins. To precisely identify the cause and the place of the pathway deregulation, it is necessary to identify phosphorylation states and concentrations of several proteins located at different levels of the regulatory cascade. In the present study, we propose the simultaneous use of specific antibodies conjugated with different quantum dots to highlight the nature of AKT/PKB cascade deregulation in patients with colorectal cancer and the loss of PTEN expression in tumor tissue. Fifty patients with colorectal cancer of no specific location were enrolled in the study. The expression of the PTEN protein, and concentrations of phosphorylated/activated forms of 3-Phosphoinositide-dependent kinase 1 (PDK1) and AKT were assessed using quantum dot-conjugated antibodies. In patients with a diminished or complete loss of the PTEN expression in the tumor tissue increased levels of activated/phosphorylated forms of PDK1 (Phospho-PDK1-Ser241) and AKT (Phospho-AKT-Thr308) proteins were found, which are responsible for the permanent activation of the phosphoinositide 3-kinase/AKT/PTEN signaling pathway in certain cases of colorectal cancer.

Effective treatment of solitary rectal ulcer syndrome using argon plasma coagulation

Solitary rectal ulcer syndrome (SRUS) is a chronic, multiform, non-cancerous disorder of the rectum, the final diagnosis of which is based upon histopathological criteria. This disorder is often accompanied by latent proctoptosis. We present a patient who (in 1996) was the first case in which argon plasma coagulation (APC) was used for SRUS treatment. In the years 2004–2005 the same patient underwent 15 APC sessions (at monthly intervals) obtaining full recovery from SRUS, although she had been treated unsuccessfully for 17 years prior to that. Six-year observation did not show any relapse. Local therapy with APC seems to be an important alternative in SRUS treatment without prolapse of the rectum and could become a basic method for bleeding treatment in SRUS.

Profile of Expression of Genes Encoding Matrix Metallopeptidase 9 (MMP9), Matrix Metallopeptidase 28 (MMP28) and TIMP Metallopeptidase Inhibitor 1 (TIMP1) in Colorectal Cancer: Assessment of the Role in Diagnosis and Prognostication

Background Studies on the pathomechanism of colorectal cancer (CRC) expansion indicate a significant role of metalloproteinases and their inhibitors in the extracellular matrix. The results of the analysis of a profile of transcriptional activity of genes encoding metalloproteinases were the basis of the hypothesis indicating changes in the expression of genes encoding MMP9, MMP28, and TIMP1 as an additional diagnostic and prognostic marker of CRC. Material/Methods The material consisted of samples obtained from resected tumors and healthy tissue samples from 15 CRC patients (aged 46–72 years) at clinical stages (CSs) I and II–IV. Gene expression analysis was done using microarrays. Microarray data analysis was done using the GeneSpring 11.5 platform. The results were validated using the qRT-PCR technique. Results We found high levels of expression of MMP9 at each CS, as well as in the tissues at the early stage of CRC. Additionally, we observed high levels of expression of TIMP1 and low levels of MMP28 genes in CS II–IV. No statistically significant differences based on the stage of CRC were observed. Conclusions MMP9 gene profile may be a complementary diagnostic marker in CRC. The results suggest a crucial role of MMP9 at the early stage of carcinogenesis in the large intestine. The increase in MMP9 and TIMP1 mRNA concentration and the decrease in MMP28 in the large intestinal tissue may be a confirmation of cancer, but it may not indicate the advance of CRC.

Direct MRI Fistulography with Hydrogen Peroxide in Patients with Recurrent Perianal Fistulas: A New Proposal of Extended Diagnostics

Background Perianal fistulas are malformations of the anorectal area. Accurate preoperative assessment of perianal fistula tract is a main assumption in diagnosis of the disease, affecting the operation efficiency. The aim of the study was to present our experience in application of a new diagnostic protocol based on the magnetic resonance imaging (MRI) examination using a mixture of hydrogen peroxide (HP) and gadolinium as a direct contrast medium in evaluation of recurrent fistulas tract. The method is referred to as HPMRI. Material/Methods The study group consisted of 12 subjects operated on from 2011. Direct HPMRI fistulography was performed in all subjects before the operation. All types of fistulas were precisely evaluated by HPMRI examination. Results Intraoperative state confirmed complete course of fistulas in 11 cases. In 1 case, an internal opening was not found. Conclusions We suggest that this new method of direct HPMRI fistulography may improve visualization of the tracts of recurrent fistulas and improve efficacy of surgical procedures.

Expression Level of Genes Coding for Cell Adhesion Molecules of Cadherin Group in Colorectal Cancer Patients.

Background Colorectal Cancer (CRC) is one of the most frequently diagnosed neoplasms and also one of the main death causes. Cell adhesion molecules are taking part in specific junctions, contributing to tissue integrality. Lower expression of the cadherins may be correlated with poorer differentiation of the CRC, and its more aggressive phenotype. The aim of the study is to designate the cadherin genes potentially useful for the diagnostics, prognostics, and the treatment of CRC. Material/Method Specimens were collected from 28 persons (14 female and 14 male), who were operated for CRC. The molecular analysis was performed using oligonucleotide microarrays, mRNA used was collected from adenocarcinoma, and macroscopically healthy tissue. The results were validated using qRT-PCR technique. Results Agglomerative hierarchical clustering of normalized mRNA levels has shown 4 groups with statistically different gene expression. The control group was divided into 2 groups, the one was appropriate control (C1), the second (C2) had the genetic properties of the CRC, without pathological changes histologically and macroscopically. The other 2 groups were: LSC (Low stage cancer) and HSC (High stage cancer). Consolidated results of the fluorescency of all of the differential genes, designated two coding E-cadherin (CDH1) with the lower expression, and P-cadherin (CDH3) with higher expression in CRC tissue. Conclusions The levels of genes expression are different for several groups of cadherins, and are related with the stage of CRC, therefore could be potentially the useful marker of the stage of the disease, also applicable in treatment and diagnostics of CRC.