Urinary Measurement of Epigenetic DNA Modifications and 8-oxodG as Possible Noninvasive Markers of Colon Cancer Evolution.

The active DNA demethylation mechanism involves 5-methylcytosine (5-mCyt) enzymatic oxidation with the subsequent formation of 5-hydroxymethylcytosine (5-hmCyt), which can be further oxidized to 5-formylcytosine (5-fCyt) and 5-carboxylcytosine (5-caCyt). The products of active DNA demethylation are released into the bloodstream and eventually also appear in urine. We used online two-dimensional ultraperformance liquid chromatography with tandem mass spectrometry (2D-UPLC-MS/MS) to compare DNA methylation marks and 8-oxo-2'-deoxyguanosine (8-oxodG) in colorectal cancer and pre-cancerous condition in urine. The study included four groups of subjects: healthy controls, patients with inflammatory bowel disease (IBD), persons with adenomatous polyps (AD), and individuals with colorectal cancer (CRC). We have found that the level of 5-fCyt in urine was significantly lower for CRC and polyp groups than in the control group. The level of 5-hmCyt was significantly higher only in the CRC group compared to the control (2.3 vs. 2.1 nmol/mmol creatinine). Interestingly, we have found highly statistically significant correlation of 5-hydroxymethyluracil with 5-hydroxymethylcytosine, 5-(hydroxymethyl)-2'-deoxycytidine, 5-(hydroxymethyl)-2'-deoxyuridine, and 5-methyl-2'-deoxycytidine in the CRC patients' group.

Body composition among patients undergoing surgery for colorectal cancer.

INTRODUCTION: Nutritional status assessment is an important part of preoperative patient evaluation, but the standard anthropometric parameters do not appear to be adequate. AIM: To determine the changes in the values of bioelectrical impedance analysis (BIA) parameters in patients 3 months after undergoing surgery for colorectal cancer (CRC). MATERIAL AND METHODS: BIA and nutritional status assessment parameters were determined in 80 patients prior to undergoing surgery for CRC. The results 3 months after surgery for 64 of those patients were then compared with their initial assessments. RESULTS: According to standard WHO ranges, 54% of the patients were diagnosed as being overweight and 29% as obese. The percentage of patients categorized as obese amounted to 56% when this was defined as high fat mass. Moderate sarcopaenia, defined as a low skeletal muscle index (SMI) or low percentage of skeletal muscle mass, was diagnosed in 21% and 29% of patients, respectively. Patients with postoperative weakness that made it impossible for them to attend the control visit had a lower preoperative skeletal muscle mass (p = 0.01) and SMI value (p = 0.001). Parameters of BIA did not discriminate patients with postoperative complications, which occurred in 23% of individuals enrolled. CONCLUSIONS: A significant proportion of the patients undergoing surgery for CRC were overweight or obese, which could mask the sarcopaenia that presented in 21-29% of them. Sarcopaenia was the only parameter predictive of a postoperative decrease in performance status.

Circulating Leptin, Adiponectin, and Tumor Necrosis Factor-Alpha in Patients Undergoing Surgery Due to Colorectal Cancer.

BACKGROUND: Adipocytokines have been proposed as factors mediating associations between obesity and inflammation in patients with colorectal cancer (CRC). Thus, the aim of this study was to determine the clinical relationships between blood concentrations of leptin (LEP), adiponectin (ADP), and tumor necrosis factor alpha (TNF-alpha) and the outcomes measured in patients with CRC undergoing surgery. PATIENTS AND METHODS: History, body composition, and blood concentrations of LEP, ADP, and TNF-alpha were determined in 107 patients undergoing surgery due to CRC. The patients were followed up for 619.72 ± 371.65 days. RESULTS: Compared to patients with stage II CRC, individuals with clinical stage I CRC had significantly lower ADP and higher TNF-alpha blood concentrations. We found significant correlations between the clinical stage of CRC (early vs. localized vs. metastatic) and the following: crude blood ADP concentration (R = 0.25; p = 0.015), ADP-to-TNF-alpha ratio (R = 0.31; p = 0.002), and ADP when indexed to body surface area (R = 0.25; p = 0.008) and to fat mass (R = 0.25; p = 0.016). The risk of death during the long-term follow-up period was independently related to the clinical stage of CRC, impairment of the patient's functional status, and higher blood carcinoembryonic antigen concentration. In Kaplan-Meier survival analysis, patients with blood LEP concentrations adjusted to a visceral adipose tissue score of ≥0.47 had a significantly better likelihood of surviving than their counterparts. CONCLUSIONS: In patients with CRC undergoing surgery, blood ADP and TNF-alpha concentrations were associated with the clinical stage of the cancer, likelihood of radical tumor excision, occurrence of nonsurgical postoperative complications, and long-term survival, which suggests the role of dysregulation in the endocrine function of adipose tissue in response to the neoplasmatic process.

Total blood lymphocyte count as a prognostic factor among unselected inpatients.

PURPOSE: The aim of this study was to determine the relationships between total lymphocyte count (TLC) and prognosis among inpatients. PATIENTS AND METHODS: We retrospectively analyzed data from electronic medical documentation of 54 976 inpatients hospitalized in an urban university hospital during 3 consecutive years (2014-2017). RESULTS: TLC was available for 12 651 (23.01%) of the inpatients. Patients with TLC <0.8 G/L constituted about 15% of the inpatients studied and had the highest risk of death, hospital readmission within 14 days, hospital readmission within 30 days and hospital readmission within 1 year, the lowest values for biochemical parameters of nutritional status assessment, and the highest C-reactive protein levels. An increase in TLC was associated with reduced risk of in-hospital death (odds ratio [OR]; 95% confidence interval [CI]): 0.31; 0.27-0.36 and 14-day readmission: 0.78; 0.72-0.86. The risk of in-hospital death associated with the Nutritional Risk Screening 2002 (NRS-2002) score, blood albumin concentration, and the score for the combined values of hemoglobin, TLC, albumin and neutrophils (HLAN) was (OR; 95% CI): 2.44; 2.35-2.53; 0.32; 0.28-0.36; and 0.96; 0.94-0.97; respectively. CONCLUSIONS: TLC < 0.8 G/L is associated with the highest risk of in-hospital death, 14-day and 30-day readmission, and longer in-hospital stay. An increase in TLC predicted in-hospital survival and freedom from early readmission with a power similar to or greater than a number of prognostic formulas based on questionnaires (e.g. NRS-2002), biochemical parameters (e.g. albumin) and formulas composed of multiple parameters (e.g. HLAN).

Blood albumin as a prognostic factor among unselected medically treated inpatients.

Aim: The aim of this study is to determine the prognostic value of blood albumin (BA) in an unselected population of inpatients. Materials & methods: We performed prospective analysis of the medical documentation of 7279 patients hospitalized between July 2014 and September 2017. Results: Individuals with BA ≥3.35 mg/dl had significantly lower risk of in-hospital death (odds ratio [OR]: 0.22; 95% CI: 0.19-0.27; p < 0.001) and 14-day readmission (OR: 0.64; 95% CI: 0.55-0.77; p < 0.0001). BA concentration was the strongest favorable factor predicting inpatient survival in a Cox hazard regression model (OR: 0.43; 95% CI: 0.36-0.50; p < 0.001), did not correlate with body mass index and actual-to-ideal bodyweight ratio and was strongly affected by numerous non-nutrient factors. Conclusion: BA concentration showed similar or better predictive and diagnostic power in relation to all-cause in-hospital mortality and 14-day readmission among inpatients than selected multifactorial scores.

Body mass index as a biomarker for the evaluation of the "Obesity Paradox" among inpatients.

BACKGROUND: Overweight and obesity are, on the one hand, recognized as risk factors for many health-related disorders, and, on the other, as favorable prognostic factors in various patients treated for several different conditions; what is called the "obesity paradox". Until now, the existence of this phenomenon among a general population of consecutive inpatients has not been evaluated. We decided, therefore, to perform an evaluation. PATIENTS AND METHODS: Historical prospective analysis of the medical documentation of 23 603 hospitalizations during two consecutive years in one center was performed. The outcomes measured were as follows: length of stay, in-hospital all-cause mortality, and non-scheduled readmission in the 14-day, 30-day and one-year periods following discharge. RESULTS: Overweight and obese patients had a lower or similar prevalence of the measured outcomes than malnourished patients and those of normal weight. Adjustment of the standard WHO BMI ranges for patients aged ≥65 y (normal weight BMI range 23-33 kg/m2) made these differences more apparent. In logistic regression, the ratio of fat to fat-free body mass was a stronger and unfavorable risk factor compared with BMI for the measured outcomes. CONCLUSIONS: The greatest risk of all-cause in-hospital death and readmission concerned malnourished inpatients. Compared to patients with a normal BMI range, overweight and obesity had a lower or similar (but not greater) risk of the outcomes measured. However, due to several BMI limitations, our observations should be interpreted as suggesting a "BMI paradox", rather than an "obesity paradox".

In vivo evidence of ascorbate involvement in the generation of epigenetic DNA modifications in leukocytes from patients with colorectal carcinoma, benign adenoma and inflammatory bowel disease.

BACKGROUND: A characteristic feature of malignant cells, such as colorectal cancer cells, is a profound decrease in the level of 5-hydroxymethylcytosine, a product of 5-methylcytosine oxidation by TET enzymes. Recent studies showed that ascorbate may upregulate the activity of TET enzymes in cultured cells and enhance formation of their products in genomic DNA. METHODS: The study included four groups of subjects: healthy controls (n = 79), patients with inflammatory bowel disease (IBD, n = 51), adenomatous polyps (n = 67) and colorectal cancer (n = 136). The list of analyzed parameters included (i) leukocyte levels of epigenetic DNA modifications and 8-oxo-7,8-dihydro-2'-deoxyguanosine, a marker of oxidatively modified DNA, determined by means of isotope-dilution automated online two-dimensional ultra-performance liquid chromatography with tandem mass spectrometry, (ii) expression of TET mRNA measured with RT-qPCR, and (iii) chromatographically-determined plasma concentrations of retinol, alpha-tocopherol and ascorbate. RESULTS: Patients from all groups presented with significantly lower levels of 5-methylcytosine and 5-hydroxymethylcytosine in DNA than the controls. A similar tendency was also observed for 5-hydroxymethyluracil level. Patients with IBD showed the highest levels of 5-formylcytosine and 8-oxo-7,8-dihydro-2'-deoxyguanosine of all study subjects, and individuals with colorectal cancer presented with the lowest concentrations of ascorbate and retinol. A positive correlation was observed between plasma concentration of ascorbate and levels of two epigenetic modifications, 5-hydroxymethylcytosine and 5-hydroxymethyluracil in leukocyte DNA. Moreover, a significant difference was found in the levels of these modifications in patients whose plasma concentrations of ascorbate were below the lower and above the upper quartile for the control group. CONCLUSIONS: These findings suggest that deficiency of ascorbate in the blood may be a marker of its shortage in other tissues, which in turn may correspond to deterioration of DNA methylation-demethylation. These observations may provide a rationale for further research on blood biomarkers of colorectal cancer development.

Characteristic profiles of DNA epigenetic modifications in colon cancer and its predisposing conditions-benign adenomas and inflammatory bowel disease.

Background: Active demethylation of 5-methyl-2'-deoxycytidine (5-mdC) in DNA occurs by oxidation to 5-(hydroxymethyl)-2'-deoxycytidine (5-hmdC) and further oxidation to 5-formyl-2'-deoxycytidine (5-fdC) and 5-carboxy-2'-deoxycytidine (5-cadC), and is carried out by enzymes of the ten-eleven translocation family (TETs 1, 2, 3). Decreased level of epigenetic DNA modifications in cancer tissue may be a consequence of reduced activity/expression of TET proteins. To determine the role of epigenetic DNA modifications in colon cancer development, we analyzed their levels in normal colon and various colonic pathologies. Moreover, we determined the expressions of TETs at mRNA and protein level.The study included material from patients with inflammatory bowel disease (IBD), benign polyps (AD), and colorectal cancer (CRC). The levels of epigenetic DNA modifications and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in examined tissues were determined by means of isotope-dilution automated online two-dimensional ultraperformance liquid chromatography with tandem mass spectrometry (2D-UPLC-MS/MS). The expressions of TET mRNA were measured with RT-qPCR, and the expressions of TET proteins were determined immunohistochemically. Results: IBD was characterized by the highest level of 8-oxodG among all analyzed tissues, as well as by a decrease in 5-hmdC and 5-mdC levels (at a midrange between normal colon and CRC). AD had the lowest levels of 5-hmdC and 5-mdC of all examined tissues and showed an increase in 8-oxodG and 5-(hydroxymethyl)-2'-deoxyuridine (5-hmdU) levels. CRC was characterized by lower levels of 5-hmdC and 5-mdC, the lowest level of 5-fdC among all analyzed tissues, and relatively high content of 5-cadC. The expression of TET1 mRNA in CRC and AD was significantly weaker than in IBD and normal colon. Furthermore, CRC and AD showed significantly lower levels of TET2 and AID mRNA than normal colonic tissue. Conclusions: Our findings suggest that a complex relationship between aberrant pattern of DNA epigenetic modification and cancer development does not depend solely on the transcriptional status of TET proteins, but also on the characteristics of premalignant/malignant cells. This study showed for the first time that the examined colonic pathologies had their unique epigenetic marks, distinguishing them from each other, as well as from normal colonic tissue. A decrease in 5-fdC level may be a characteristic feature of largely undifferentiated cancer cells.

Characteristics and results of treatment of patients treated surgically with colorectal cancer in old and senile age.

Colorectal cancer (CC) in Poland is the type of cancer with the highest dynamics of disease growth and is epidemiologically related to age. The analysis involved 353 patients operated on due to CC in senile and old age and compared with younger patients. It was found that people at this age are more often diagnosed with CC They were more often women, the patients did not differ in the stage of cancer, while they were significantly more often qualified for surgery due to urgent indications. In patients with colonic cancer, the resectability and radicality of the procedures in comparison with patients with rectal cancer was significantly higher, while there were more complications and deaths in the 30-day follow-up in this group. The overall survival in senile and old age was significantly worse. In the first year of follow-up after surgical treatment of patients in this group, complications and deaths were more frequently observed. However, in patients who survived 12 months after the operation, the overall survival rate did not significantly differ.

Mitogenomic differences between the normal and tumor cells of colorectal cancer patients.

So far, a reliable spectrum of mitochondrial DNA mutations in colorectal cancer cells is still unknown, and neither is their significance in carcinogenesis. Indeed, it remains debatable whether mtDNA mutations are "drivers" or "passengers" of colorectal carcinogenesis. Thus, we analyzed 200 mitogenomes from normal and cancer tissues of 100 colorectal cancer patients. Minority variant mutations were detected at the 1% level. We showed that somatic mutations frequently occur in colorectal cancer cells (75%) and are randomly distributed across the mitochondrial genome. Mutational signatures of somatic mitogenome mutations suggest that they might arise through nucleotide deamination due to oxidative stress. The majority of somatic mutations localized within the coding region (in positions not known from the human phylogeny) and was potentially pathogenic to cell metabolism. Further analysis suggested that the relaxation of negative selection in the mitogenomes of colorectal cancer cells may allow accumulation of somatic mutations. Thus, a shift in glucose metabolism from oxidative phosphorylation to glycolysis may create advantageous conditions for accumulation of mtDNA mutations. Considering the fact that the presence of somatic mtDNA mutations was not associated with any clinicopathological features, we suggested that mtDNA somatic mutations are "passengers" rather than the cause of colorectal carcinogenesis.

The "cholesterol paradox" among inpatients - retrospective analysis of medical documentation.

INTRODUCTION: There is evidence of positive relationships between cholesterol concentration and risk of cardiovascular diseases. However, higher mortality in patients with a low cholesterol level has been reported (the "cholesterol paradox"). MATERIAL AND METHODS: Medical records of 34 191 inpatients between 2014 and 2016 were reviewed and the relationships between total (TC), low-density lipoprotein (LDL-C) and high-density lipoprotein (HDL-C) cholesterol and triglyceride blood concentrations and all-cause in-hospital death and readmission within 14 and 30 days and 1 year were determined in univariate and multivariate analyses. RESULTS: Patients with TC in the lower quartile and LDL-C < 70 mg/dl had greater risk of the outcomes measured than individuals with a TC level in the remaining quartiles and LDL-C ≥ 70 mg/dl. Moreover, patients with TC in the highest quartile, OR (95% CI): 0.36 (0.13-0.99), p < 0.05, and LDL-C ≥ 115 mg/dl, OR (95% CI): 0.53 (0.37-0.77), p < 0.05, had the lowest all-cause in-hospital mortality. However, multivariate analysis using logistic regression and a Cox proportional hazard model showed no significant influence of blood lipid levels on the occurrence of the outcomes measured. CONCLUSIONS: A significant effect of a "cholesterol paradox" linking better prognosis with higher blood lipid concentration was found only in univariate analysis but, after adjustment for clinical characteristics in multivariate analysis, the plasma lipid level had a neutral influence on the occurrence of the measured outcomes. This suggests that a low cholesterol level should be interpreted as a biomarker of illness severity.

Surgical site infection among patients after colorectal cancer surgery.

Wstep: Zakazenie miejsca operowanego wystepuje u 2,5-22,3% operowanych chorych. Jest ono wykladnikiem jakosci leczenia na oddzialach zabiegowych i ma duzy wplyw na jego koszt. Material i metodyka: Analizie poddano chorych, u których w obserwacji 30-dniowej wystapilo zakazenie miejsca operowanego. Grupe wyjsciowa stanowilo 1581 chorych z rozpoznaniem raka jelita grubego poddanych zabiegowi operacyjnemu w jednym osrodku. Kryteriami wylaczajacymi z badania byly: brak wiarygodnej dokumentacji leczenia (szpitalnego lub ambulatoryjnego) i zgon chorego przed 30. dniem po operacji bez rozpoznanego zakazenia miejsca operowanego. Analize statystyczna wykonano przy uzyciu programu Statistica 10. Wyniki: Powiklania pooperacyjne wystapily u 262 chorych (16,6%). Najczesciej wystepujacym bylo zakazenie miejsca operowanego (198 pacjentów; 12,52%). Stwierdzono, ze wystapienie tego powiklania zalezne bylo od zaawansowania klinicznego raka, wieku chorych, chorób wspóltowarzyszacych (cukrzyca i choroby kardiologiczne). Ponadto zauwazono, ze powiklanie to wystepowalo znamiennie czesciej u chorych operowanych w trybie pilnym z powodu powiklan oraz u tych, u których wyloniono stomie jelitowa. Nie stwierdzono natomiast zaleznosci wystapienia tego powiklania od plci chorych i lokalizacji guza nowotworowego. Wniosek: U chorych po operacji raka jelita grubego najwieksze zagrozenie wystapienia zakazenia miejsca operowanego wystapilo u chorych po 75. roku zycia, obciazonych cukrzyca i chorobami kardiologicznymi, z duzym zaawansowaniem klinicznym raka, operowanych w trybie ostrego dyzuru, u których konieczne bylo wylonienie stomii jelitowej (a szczególnie kolostomii).

The fecal occult blood test as a tool for improved outpatient qualification for colonoscopy. A single-center experience and 10-year follow-up survey.

PURPOSE: Colonoscopy is not widely and easily available in all countries, even for symptomatic patients. This is one of the causes of tumors not being diagnosed until an advanced stage. The aim of this study was to estimate the efficacy of the fecal occult blood test (FOBT) in the diagnostic work-up of outpatients referred to a colorectal unit due to indistinct abdominal symptoms. PATIENTS/METHODS: Among 10418 consecutive symptomatic individuals referred to the outpatient clinic, an immunochemical FOBT (Hem-Check 1®) was recommended for 9432 patients with indistinct symptoms as a tool for qualifying them for colonoscopy. All the subjects were treated according to their diagnosis and followed-up for the next 10 years. RESULTS: Colorectal cancer (CRC) was diagnosed in 535 individuals: 393/986 (39.9%) among patients with red-flag symptoms, and 142/951 (14.9%) of individuals with indistinct symptoms and a positive FOBT. In the latter group, less-advanced tumors, classed as such using Dukes' classification, were twice as common and more advanced CRC occurred twice as seldom than in the former. Cancer recurrence-free and overall survival periods after surgical treatment for CRC were significantly longer in patients with indistinct symptoms who qualified for diagnostic procedures on the basis of a positive FOBT. CONCLUSIONS: Patients with symptoms suggesting organic colon disease had a worse prognosis compared to individuals with non-specific symptoms. If bowel endoscopy is not widely and easily available, qualification for colonoscopy on the basis of alarm symptoms and a positive FOBT seems to be an effective strategy in early CRC diagnosis.

Urinary Measurement of Epigenetic DNA Modifications: A Non-Invasive Assessment of the Whole-Body Epigenetic Status in Healthy Subjects and Colorectal Cancer Patients.

Active mechanism of DNA demethylation can be responsible for the activation of previously silenced genes. Products of 5-methylcytosine oxidation are released into the bloodstream and eventually excreted with urine. Therefore, whole-body epigenetic status can be assessed non-invasively on the basis of the urinary excretion of a broad spectrum of epigenetic modifications: 5-hydroxymethylcytosine (5-hmCyt), 5-formylcytosine (5-fCyt), 5-carboxycytosine (5-caCyt), and 5-hydroxymethyluracil (5-hmUra). We have developed a specific and sensitive, isotope-dilution, automated, online, two-dimensional ultra-performance liquid chromatography system with tandem mass spectrometry (2D UPLC-MS/MS) to measure 5-hmCyt, 5-fCyt, 5-caCyt, and their deoxynucleosides in the same urine sample. Human urine contains all of the modifications except from 5-formyl-2'-deoxycytidine (5-fdC) and 5-carboxy-2'-deoxycytidine (5-cadC). A highly significant difference in the urinary excretion of 5-(hydroxymethyl)-2'-deoxycytidine (5-hmdC) was found between healthy subjects and colorectal cancer patients (3.5 vs. 7.8 nmol mmol-1 creatinine, respectively), as well as strong correlations between the majority of analyzed compounds.

Scores of nutritional risk and parameters of nutritional status assessment as predictors of in-hospital mortality and readmissions in the general hospital population.

BACKGROUND & AIMS: We have no "gold standard" for the diagnosis of malnutrition. The aim of this study was to determine the importance of many of the parameters used in nutritional status screening and assessment among inpatients for the prediction of in-hospital mortality, readmission and length of hospitalization. METHODS: On the base of the medical documentation a retrospective analysis was performed of nutritional status screening and assessment parameters for all 20,237 non-selected, consecutive hospitalizations in 15,013 patients over 18 years of age treated in one hospital during the course of one year. RESULTS: The risk of malnutrition expressed as a Nutritional Risk Screening (NRS)-2002 score ≥ 3 concerned 6.4% hospitalizations. The greater risk of in-hospital death, as well as readmission within 14 days and 30 days, was related to an NRS-2002 score ≥3, age >65 years, male gender, urgent admission, body mass deficit calculated as the difference between actual body mass and ideal weight determined according to the Lorentz formula, higher degree of Instant Nutritional Assessment (INA), greater value of a C-reactive protein (CRP)/albumin ratio, and plasma glucose concentration. Whereas, greater blood concentration of albumin, hemoglobin, cholesterol and triglycerides, as well as a greater blood lymphocyte count, were associated with reduced risk of the measured outcomes. CONCLUSIONS: NRS-2002 score, blood albumin, CRP/albumin ratio, and INA seem to be good predictors of in-hospital mortality, readmission rate and length of hospital stay.

Accurate, Direct, and High-Throughput Analyses of a Broad Spectrum of Endogenously Generated DNA Base Modifications with Isotope-Dilution Two-Dimensional Ultraperformance Liquid Chromatography with Tandem Mass Spectrometry: Possible Clinical Implication.

Our hereby presented methodology is suitable for reliable assessment of the most common unavoidable DNA modifications which arise as a product of fundamental metabolic processes. 8-Oxoguanine, one of the oxidatively modified DNA bases, is a typical biomarker of oxidative stress. A noncanonical base, uracil, may be also present in small quantities in DNA. A set of ten-eleven translocation (TET) proteins are involved in oxidation of 5-methylcytosine to 5-hydroxymethylcytosine which can be further oxidized to 5-formylcytosine and 5-carboxycytosine. 5-Hydroxymethyluracil may be formed in deamination reaction of 5-hydroxymethylcytosine or can be also generated by TET enzymes. All of the aforementioned modifications seem to play some regulatory roles. We applied isotope-dilution automated online two-dimensional ultraperformance liquid chromatography with tandem mass spectrometry (2D-UPLC-MS/MS) for direct measurement of the 5-methyl-2'-deoxycytidine, 5-(hydroxymethyl)-2'-deoxycytidine, 5-formyl-2'-deoxycytidine, 5-carboxy-2'-deoxycytidine, 5-(hydroxymethyl)-2'-deoxyuridine, 2'-deoxyuridine, and 8-oxo-2'-deoxyguanosine. Analyses of DNA extracted from matched human samples showed that the 5-(hydroxymethyl)-2'-deoxycytidine level was 5-fold lower in colorectal carcinoma tumor in comparison with the normal one from the tumor's margin; also 5-formyl-2'-deoxycytidine and 5-carboxy-2'-deoxycytidine were lower in colorectal carcinoma tissue (ca. 2.5- and 3.5-fold, respectively). No such differences was found for 2'-deoxyuridine and 5-(hydroxymethyl)-2'-deoxyuridine. The presented methodology is suitable for fast, accurate, and complex evaluation of an array of endogenously generated DNA deoxynucleosides modifications. This novel technique could be used for monitoring of cancer and other diseases related to oxidative stress, aberrant metabolism, and environmental exposure. Furthermore, the fully automated two-dimensional separation is extremely useful for analysis of material containing a considerable amount of coeluting interferents with mass-spectrometry-based methods.

Urinary 5-hydroxymethyluracil and 8-oxo-7,8-dihydroguanine as potential biomarkers in patients with colorectal cancer.

CONTEXT: Oxidative stress linked with chronic inflammation is associated with etiology of the colorectal cancer. OBJECTIVES: To assess the diagnostic utility of urinary excretion of oxidatively modified DNA bases/nucleoside: 8-oxo-7,8-dihydroguanine (8-oxoGua), 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) and 5-hydroxymethyluracil (5-hmUra). MATERIALS AND METHODS: Seventy-two healthy controls, 15 patients with adenomas and 56 colorectal cancer patients were recruited. RESULTS: The receiver operating characteristic (ROC) curve analysis showed that the area under the curve (AUC) for all markers tested separately was <0.7. The combination of these modifications showed better diagnostic power (AUC = 0.778 for 8-oxoGua + 8-oxodG)/5hmUra ratio). CONCLUSION: Urinary DNA modifications may reflect the oxidative stress/chronic inflammation in colorectal cancer but diagnostic performance for early-detection is moderate.

Cumulative Small Effect Genetic Markers and the Risk of Colorectal Cancer in Poland, Estonia, Lithuania, and Latvia.

The continued identification of new low-penetrance genetic variants for colorectal cancer (CRC) raises the question of their potential cumulative effect among compound carriers. We focused on 6 SNPs (rs380284, rs4464148, rs4779584, rs4939827, rs6983267, and rs10795668), already described as risk markers, and tested their possible independent and combined contribution to CRC predisposition. Material and Methods. DNA was collected and genotyped from 2330 unselected consecutive CRC cases and controls from Estonia (166 cases and controls), Latvia (81 cases and controls), Lithuania (123 cases and controls), and Poland (795 cases and controls). Results. Beyond individual effects, the analysis revealed statistically significant linear cumulative effects for these 6 markers for all samples except of the Latvian one (corrected P value = 0.018 for the Estonian, corrected P value = 0.0034 for the Lithuanian, and corrected P value = 0.0076 for the Polish sample). Conclusions. The significant linear cumulative effects demonstrated here support the idea of using sets of low-risk markers for delimiting new groups with high-risk of CRC in clinical practice that are not carriers of the usual CRC high-risk markers.

Intestinal stoma in patients with colorectal cancer from the perspective of 20-year period of clinical observation.

INTRODUCTION: Intestinal stoma is a procedure most often performed in patients with colorectal cancer. AIM: To identify the percentage of patients with colorectal cancer in which the intestinal stoma was performed. MATERIAL AND METHODS: We retrospectively analysed 443 patients treated during a 20-year period (1994-2013) due to colorectal cancer, in which the intestinal stoma was made during the first surgical intervention. RESULTS: In the second analysed decade, a significant decrease in the percentage of created stomas, definitive stomas in particular, was observed. Stomas were made significantly more often in patients with a tumour located in the rectum, the left half of the colon, and in patients undergoing urgent surgeries. An increased incidence of intestinal stomas was associated with a higher severity of illness and higher proportion of unresectable and non-radical procedures. The definitive stomas were significantly more often made in men and in patients with tumours located in the rectum, whereas temporary stomas were created significantly more often in patients undergoing urgent operations. CONCLUSIONS: In the last decade (2004-2013) the number of intestinal stomas in patients operated due to colorectal cancer was significantly reduced.

Inflammatory myofibroblastic tumor within ileal intussusception as the cause of recurrent abdominal pain in a 57-year old patient.

The study presented a rare case of inflammatory myofibroblastic tumor (IMT) in a 57-year old male patient who presented with recurrent abdominal pain. He was diagnosed to have a tumor of the small intestine within ileal intussusception, demonstrated by CT enterography. The patient underwent surgery to remove the tumor with a margin of healthy tissue. Histopathological and immunohistochemical examination results enabled to diagnose IMT. IMT is a rare tumor that occurs mainly in children and young adults. Its etiology remains to be fully understood. Due to the differentiated histology it can be found in many organs and soft tissues, being responsible for different, non-specific clinical and radiological symptoms. Due to the rarity of this tumor a clear treatment protocol has not yet been established. However, given the tendency to recur with possible distant metastases an important element of treatment consists in long-term clinical patient observation.