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The quantity and quality of preoperative material in colorectal cancer is often limiting factor in determination of risk factors and therapy planning. The most important negative prognostic factors are intravascular and perineural invasion, as well as tumor budding. Usually, the only parameter available in preoperative biopsy is tumor budding. However, the growing body of evidence suggests that cancer differentiation based on the poorly differentiated clusters has better prognostic value. The limiting factor in applying of these new parameters is reproducible, simple, cheap and fast method of their determination. In this paper we investigated the prognostic value of lacunarity, determined in preoperative biopsy. Lacunarity is a measure of spatial heterogeneity (inhomogeneity) in an image. It quantifies how objects fill the space, and enables analysis of gaps distribution, homogeneity of gaps, and presence of structures. It was shown that lacunarity and the total number of buds could be combined in a model which clearly divides colorectal cancer patients in low, medium and high risk subgroups. The paper also points out that the quantitative numerical methods are superior to semiquantitative methods, and that individual methods should be combined using algorithms to obtain a more accurate prediction. Because the study described is designed as a pilot study, verification is needed on a larger sample of patients from independent researchers.

Background and aims: To evaluate two-dimensional shear wave elastography (2DSWE) in parallel with transient elastography (TE) for diagnosing clinically significant portal hypertension (CSPH) and high-risk varices (HRV) in patients with chronic liver disease. Patients and methods: Consecutive patients with suspicion of compensated advanced chronic liver disease (cACLD) [liver stiffness measurement (LSM) ≥ 10 kPa by TE, or morphological signs suggestive of cACLD on imaging], with no history of liver decompensation, underwent hepatic venous pressure gradient (HVPG) measurement, transjugular liver biopsy and esophagogastroduodenoscopy, which served as the reference methods for diagnosing CSPH, cACLD and HRV. All patients underwent LSM and spleen stiffness measurements (SSM) by 2DSWE and TE. Results: Seventy-six (76) patients were included (78% men, mean age 62 years, body mass index 28.3 kg/m2 , 36.8% alcoholic, 30.3% non-alcoholic fatty liver disease, 14.5% viral hepatitis). Of them, 80.3%, 69.7%, 52.6% and 22.4% had cACLD, cirrhosis, CSPH and HRV respectively. LSM performed better than SSM in diagnosing CSPH and HRV. For CSPH, AUROCs (0.926 vs. 0.866), optimal cut-offs (20.1 vs. 20.2 kPa) and sensitivity/specificity (80.5%/94.3% vs. 77.5% /86.1%) were comparable for 2DSWE and TE. Ruling-out of CSPH by 2DSWE (LSM at cut-off with ≥90% sensitivity (13.5 kPa) and platelets ≥ 150 x 109 /L) performed comparably to TE, with 1/24 cases falsely classified as negative. For HRV, AUROCs were similar (0.875 2DSWE, 0.851 TE) with similar optimal LSM cut-offs enabling 100% sensitivity and ruling-out HRV. Conclusion: Liver stiffness measurement by 2DSWE appears to perform equally well as TE for diagnosing CSPH and ruling-out HRV in compensated chronic liver disease.

The lack of pixel-wise annotated images severely hinders the deep learning approach to computer-aided diagnosis in histopathology. This research creates a public database comprised of: (i) a dataset of 82 histopathological images of hematoxylin-eosin stained frozen sections acquired intraoperatively on 19 patients diagnosed with metastatic colon cancer in a liver; (ii) corresponding pixel-wise ground truth maps annotated by four pathologists, two residents in pathology, and one final-year student of medicine. The Fleiss' kappa equal to 0.74 indicates substantial inter-annotator agreement; (iii) two datasets with images stain-normalized relative to two target images; (iv) development of two conventional machine learning and three deep learning-based diagnostic models. The database is available at http://cocahis.irb.hr. For binary, cancer vs. non-cancer, pixel-wise diagnosis we develop: SVM, kNN, U-Net, U-Net++, and DeepLabv3 classifiers that combine results from original images and stain-normalized images, which can be interpreted as different views. On average, deep learning classifiers outperformed SVM and kNN classifiers on an independent test set 14% in terms of micro balanced accuracy, 15% in terms of the micro F1 score, and 26% in terms of micro precision. As opposed to that, the difference in performance between deep classifiers is within 2%. We found an insignificant difference in performance between deep classifiers trained from scratch and corresponding classifiers pre-trained on non-domain image datasets. The best micro balanced accuracy estimated on the independent test set by the U-Net++ classifier equals 89.34%. Corresponding amounts of F1 score and precision are, respectively, 83.67% and 81.11%. The lack of pixel-wise annotated images severely hinders the deep learning approach to computer-aided diagnosis in histopathology. This research creates a public database comprised of: (i) a dataset of 82 histopathological images of hematoxylin-eosin stained frozen sections acquired intraoperatively on 19 patients diagnosed with metastatic colon cancer in a liver; (ii) corresponding pixel-wise ground truth maps annotated by four pathologists, two residents in pathology, and one final-year student of medicine. The Fleiss' kappa equal to 0.74 indicates substantial inter-annotator agreement; (iii) two datasets with images stain-normalized relative to two target images; (iv) development of two conventional machine learning and three deep learning-based diagnostic models. The database is available at http://cocahis.irb.hr. For binary, cancer vs. non-cancer, pixel-wise diagnosis we develop: SVM, kNN, U-Net, U-Net++, and DeepLabv3 classifiers that combine results from original images and stain-normalized images, which can be interpreted as different views. On average, deep learning classifiers outperformed SVM and kNN classifiers on an independent test set 14% in terms of micro balanced accuracy, 15% in terms of the micro F1 score, and 26% in terms of micro precision. As opposed to that, the difference in performance between deep classifiers is within 2%. We found an insignificant difference in performance between deep classifiers trained from scratch and corresponding classifiers pre-trained on non-domain image datasets. The best micro balanced accuracy estimated on the independent test set by the U-Net++ classifier equals 89.34%. Corresponding amounts of F1 score and precision are, respectively, 83.67% and 81.11%.

We have read with great interest manuscript by Piccinni et al. in which they demonstrated significant prevalence of liver steatosis in patients with compensated advanced chronic liver disease (cACLD), previously assumed to decrease in advanced fibrosis, as well as very good performance of Controlled attenuation parameter (CAP) in this cohort of patients (1, 2).

AIM: To compare and evaluate the hepatoprotective effect of remote ischemic preconditioning (RIPC) with local ischemic preconditioning (LIPC) of the liver during human liver resections. ----- METHODS: A prospective, single-centre, randomised control trial was conducted in the Clinical Hospital "***" from April 2017 to January 2018. A total of 60 patients, who underwent liver resection due to colorectal cancer liver metastasis, were randomised to one of three study arms: 1) a RIPC group, 2) an LIPC group and 3) a control group (CG) in which no ischemic preconditioning was done before liver resection. The hepatoprotective effect was evaluated by comparing serum transaminase levels, bilirubin levels, albumin, and protein levels, coagulograms and through pathohistological analysis. The trial was registered on ClinicalTrials.gov (NCT****). ----- RESULTS: Significant differences were found in serum levels of liver transaminases and bilirubin levels between the groups, the highest level in the CG and the lowest level in the LIPC group. Levels of cholinesterase were also significantly higher in the LIPC group. Pathohistological findings graded by the Rodriguez score showed favourable changes in the LIPC and RIPC groups versus the CG. ----- CONCLUSION: Strong evidence supports the hepatoprotective effect of RIPC and LIPC preconditioning from an ischemia-reperfusion injury of the liver. Better synthetic liver function preservation in these two groups supports this conclusion.

The aim of the study was to explore predictive value of the ALBI, PALBI and MELD scores on survival in patients resected for hepatocellular carcinoma with compensated liver cirrhosis and no macrovascular infi ltration. In this retrospective study, longitudinal survival analysis was performed. We analyzed patient/tumor characteristics and MELD, ALBI and PALBI scores as liver function tests for predicting survival outcome. Survival was analyzed from the date of liver resection until death, liver transplantation, or end of follow-up. Patients were stratifi ed for age, cirrhosis etiology, presence of esophageal varices, hepatocellular carcinoma stage, microvascular invasion, histologic diff erentiation, and resection margins. We identifi ed 38 patients (alcoholic cirrhosis in 84.2% of patients) resected over an 8-year period. Median preoperative MELD score was 8, ALBI score -2.63, and PALBI score -2.38. During the follow-up period, 24 patients died. Estimated median survival time was 36 months. Microvascular invasion was observed in 33 patients. Higher ALBI score was associated with 23.1% higher relative risk of death. PALBI score was associated with 12.1% higher relative risk of death, whereas MELD score was not associated with the risk of death. In conclusion, ALBI score demonstrated signifi cant predictive capabilities for survival in patients with compensated cirrhosis resected for hepatocellular carcinoma., Cilj je bio istražiti prediktivnu vrijednost zbira ALBI, PALBI i MELD za preživljenje bolesnika s kompenziranom cirozom reseciranih zbog hepatocelularnog karcinoma bez makrovaskularne invazije. Provedena je retrospektivna longitudinalna analiza preživljenja. Testirane su karakteristike bolesnika/tumora kao i jetreni funkcijski testovi MELD, ALBI i PALBI za predviđanje preživljenja. Bolesnici su praćeni nakon resekcije jetre do smrti, transplantacije jetre odnosno do kraja vremena praćenja. Bolesnici su kategorizirani po dobi, etiologiji ciroze, prisutnosti varikoziteta jednjaka, stadiju karcinoma, mikrovaskularnoj invaziji, histološkom stadiju i resekcijskim rubovima. Identifi cirali smo 38 bolesnika (alkoholna ciroza kod 84,2% bolenika) reseciranih kroz razdoblje od 8 godina. Medijan prijeoperacijskog zbira MELD bio je 8, zbira ALBI -2,63 i zbira PALBI -2,38. Tijekom praćenja 24 bolesnika su umrla. Očekivani medijan preživljenja iznosio je 36 mjeseci. Mikrovaskularna invazija pronađena je u 33 bolesnika. Veći ALBI je bio povezan s 23,1%, a PALBI s 12,1% većim relativnim rizikom od smrtnog ishoda. MELD nije bio povezan sa smrtnim ishodom. Zbir ALBI je pokazao značajnu prediktivnu vrijednost preživljenja u skupini bolesnika s kompenziranom cirozom reseciranih zbog hepatocelularnog karcinoma.

Background & Aims To analyse elastographic characteristics of focal liver lesions (FLL)s and diagnostic performance of real‐ time two‐dimensional shear‐wave elastography (RT‐ 2D‐SWE) in order to differentiate benign and malignant FLLs. Methods Consecutive patients diagnosed with FLL by abdominal ultrasound (US) underwent RT‐2D‐SWE of FLL and non‐ infiltrated liver by intercostal approach over the right liver lobe. The nature of FLL was determined by diagnostic work‐up, including at least one contrast‐ enhanced imaging modality (MDCT/MRI), check‐up of target organs when metastatic disease was suspected and FLL biopsy in inconclusive cases. Results We analysed 196 patients (median age 60 [range 50‐68], 50.5% males) with 259 FLLs (57 hepatocellular carcinomas, 17 cholangiocarcinomas, 94 metastases, 71 haemangiomas, 20 focal nodular hyperplasia) of which 70 (27%) were in cirrhotic liver. Malignant lesions were stiffer (P < .001) with higher variability in intralesional stiffness (P = .001). The best performing cut‐off of lesion stiffness was 22.3 kPa (sensitivity 83% ; specificity 86% ; positive predictive value [PPV] 91.5% ; negative predictive value [NPV] 73%) for malignancy. Lesion stiffness 32.5 kPa had PPV of 96% for malignancy. Lesion stiffness, lesion/liver stiffness ratio and lesion stiffness variability significantly predicted malignancy in stepwise logistic regression (P < .05), and were used to construct a new Liver Elastography Malignancy Prediction (LEMP) score with accuracy of 96.1% in validation cohort (online calculator available at http://bit.do/lemps). Conclusion The comprehensive approach demonstrated in this study enables correct differentiation of benign and malignant FLL in 96% of patients by using RT‐ 2D‐SWE.

Objectives: To investigate the clinical and prognostic significance of absolute basophil count (ABC) in patients with primary myelofibrosis (PMF). Methods: We retrospectively investigated 58 patients with PMF treated in our institution in the period from 2006 to 2017. ABC was obtained in addition to other hematological and clinical parameters. Patients were separated into high and low ABC groups using the Receiver operating characteristic curve analysis. Results: ABC was higher in PMF patients than in healthy controls (P < 0.001). Patients with high ABC had higher white blood cells (P < 0.001), higher red cell distribution width (P = 0.035), higher lactate dehydrogenase (P < 0.001), more frequently had circulatory blasts (P < 0.001), constitutional symptoms (P = 0.030) and massive splenomegaly (P = 0.014). ABC was also positively correlated with absolute monocyte count (AMC) (P < 0.001) and other components of differential blood count. There was no difference in ABC regarding driver mutations or degree of bone marrow fibrosis. Univariately, high ABC was significantly associated with inferior overall survival (hazard ratio (HR) 4.79, P < 0.001). This effect remained statistically significant (HR 4.27, P = 0.009) in a multivariate Cox regression model adjusted for age, gender, Dynamic International Prognostic Scoring System (HR 2.6, P = 0.001) and AMC (HR 8.45, P = 0.002). Discussion: High ABC reflects higher disease activity and stronger proliferative potential of disease. ABC and AMC independently predict survival and therefore seem to reflect different underlying pathophysiologic processes. Hence, both have a potential for improvement of current prognostic scores. Conclusion: Basophils represent a part of malignant clone in PMF and are associated with unfavorable disease features and poor prognosis which is independent of currently established prognostic scoring system and monocytosis.

Development of computer-aided diagnosis (CAD) systems is motivated by reduction of the workload on the pathologist that is increasing steadily. Among approaches upon which CAD-based systems are built, deep learning (DL) methods seem to be well suited for image analysis in digital pathology. However, DL networks include a large number of parameters and that requires a large annotated training dataset. Unfortunately, probably the biggest problem in digital pathology using machine learning methods is a small number of annotated images. That is especially true in intraoperative tissue analysis which coincides with the topic of the present paper: intraoperative CAD-based diagnosis of metastasis of colon cancer in a liver from hematoxylin-eosin (H and E) stained frozen section. To cope with the insufficiency of training images we adopt a transfer learning approach using the Nested UNet architecture. For better diagnostic performance, the trained model predicted pixels multiple times for different striding levels using the sliding window strategy. Threshold optimization using balanced accuracy score showed the validity of such an approach as balanced accuracy has increased significantly. When compared to often used UNet with VGG16 backbone, Nested UNet model with DenseNet201 backbone performs better on our dataset for both balanced accuracy metric and F1 score.

There is a need for computer-aided diagnosis (CAD) systems to relieve the workload on pathologists. This seems to be especially important for intraoperative diagnosis during surgery, for which diagnostic time is very limited. This paper presents preliminary results of intraoperative pixel-based CAD of colon cancer metastasis in a liver from phase- contrast images of unstained frozen sections. In particular, two deep learning networks: the U-net and the structured autoencoder for deep subspace clustering, were trained on eighteen phase-contrast images belonging to five patients and tested on eight images belonging to three patients. Spectrum angle mapper was also used in comparative performance analysis. The best result achieved by the U-net yielded balanced accuracy of 83.70%±8%, sensitivity of 94.50%±8%, specificity of 72.9%±8% and Dice coefficient of 45.20%±25.4%. However, factors such as absence of tissue fixation and ethanol- induced dehydration, melting of the specimen under the microscope and/or frozen crystals in the specimen cause variations in quality of phase-contrast images of unstained frozen sections. This, in return, affects reproducibility of diagnostic performance.

Background Despite numerous investigations, we still do not have a specific marker for pancreatic ductal adenocarcinoma. Only guideline-recommended biomarker for pancreatic ductal adenocarcinoma is the CA19-9, but it is also present in other gastrointestinal diseases. IMP3 is a new potential biomarker that is over-expressed in some cancers. The aims of our study were (1) to assess IMP3 in benign pancreatic lesions and pancreatic cancer, and (2) to estimate its concentrations in localized and advanced pancreatic cancer. Patients and methods Seventy-five patients with solid pancreatic lesions who underwent EUS-FNA were included. Patients were divided into three groups: benign lesions, cancer localized only on the pancreas, and patients with advanced pancreatic cancer (locally advanced or with distal metastases). Immunoreactivity of IMP3 was assessed on cytological smears sampled by endoscopic ultrasound. Results IMP3 was expressed in 89% of the patients with pancreatic cancer and not in benign lesions. Stronger expression of IMP3 protein and stage of the pancreatic cancer was statistically significant. IMP3 was expressed in all localized cancers and in 85% of patients with advanced pancreatic cancer. In the subgroup with locally advanced cancer, IMP3 was expressed in 88%, and in 83% of patients in the subgroup with distal metastasis (P = 0.007). In the present study, sensitivity was 89%, specificity 100%, with positive predictive value of 100% and negative predictive value of 63%. Conclusion There is a positive correlation between IMP3 expression and TNM stages of the pancreatic cancer. Higher expression of IMP3 on EUS-FNA specimens can suggest poorer prognosis.

The purpose of this paper was to outline the development of short peptide targeting of the human prostate specific antigen (hPSA), and to evaluate its effectiveness in staining PSA in human prostate cancer tissue. The targeting of the hPSA antigen by means of antisense peptide AVRDKVG was designed according to a three-step method involving: 1. The selection of the molecular target (hPSA epitope), 2. the modeling of an antisense peptide (paratope) based on the epitope sequence, and 3. the spectroscopic evaluation of sense&ndash, antisense peptide binding. We then modified standard hPSA immunohistochemical staining practice by using a biotinylated antisense peptide instead of the standard monoclonal antibody and compared the results of both procedures. Immunochemical testing on human tissue showed the applicability of the antisense peptide technology to human molecular targets. This methodology represents a new approach to deriving peptide ligands and potential lead compounds for the development of novel diagnostic substances, biopharmaceuticals and vaccines.

Background/ Aim: Thrombin plays significant roles in various types of cancer. However, the expression levels of prothrombin, the thrombin precursor, in cancer remain unclear. Variants of the 3'end of the prothrombin gene lead to increased prothrombin expression. This study aimed to analyze prothrombin 3'end gene variants in colon tumor and adjacent normal tissue samples. Materials and Methods: The study group consisted of 93 patients suffering from colon adenocarcinoma. The 3'end of the prothrombin gene was analyzed by DNA sequencing. Results: Three variants, all previously associated with increased prothrombin expression were detected. Frequency of the FII 19911G allele was 46.77% and 47.85% in tumor and normal tissue, respectively. For the FII 20210A allele, the detected frequencies were 2.15% and 1.61%, respectively. The frequency of the FII c.1824T allele was 0.54% in both tissues. Four patients showed different genotypes in tumor and normal tissue. Conclusion: Prothrombin 3' end gene variants may play a role in colorectal cancer.

Survivin, encoded by BIRC5 gene (baculoviral IAP repeat containing 5), belongs to the family of inhibitors of apoptosis proteins (IAPs). In mammalian cells it participates in the control of mitosis, apoptosis regulation, and cellular stress response. Its expression is increased in almost all types of cancers. The aim of this study was to investigate the role of BIRC5 polymorphisms in breast cancer (BC) and to connect survivin expression with various clinicopathological characteristics of BC patients. Blood and archival tumour tissue samples were collected from 26 BC patients from Croatia. Survivin expression was determined immunohistochemically. BIRC5 promoter, coding region, and 3’UTR were genotyped. DNA from 74 healthy women was used as control. BIRC5 polymorphisms and survivin expression were tested against age of onset, histological grade, tumour type and size, lymph node status, oestrogen, progesterone, Her2, and Ki67 status. Numbers of samples with weak, moderate, and strong survivin expression were 9 (33.3%), 11 (40.7%), and 7 (25.9%), respectively. Most patients had nuclear survivin staining (92.6%). High survivin expression was significantly associated with negative oestrogen receptor status (p=0.007) and positive Ki67 expression (p=0.032). Ki67 expression was also positively correlated with histological grade (p=0.0009). Fourteen polymorphisms were found in BC samples, located mostly in promoter and 3’UTR of BIRC5. There was no significant difference in the distribution of polymorphisms between BC and control samples. Among clinicopathological characteristics of BC patients, alleles of five BIRC5 polymorphisms were associated with younger age of onset: c.-644T>C (55.8 years [y] vs. 48.1 y; p=0.006), c.-241C>T (54.2 y vs. 45.0; p=0.029), c.9809T>C (55.8 y vs. 48.1 y; p=0.006), c.-1547C>T (58.3 y vs. 50.9 y; p=0.011), and c.9386T>C (50.8 y vs. 59.5 y; p=0.004). To assess the significance of BIRC5 polymorphisms and survivin expression as predictive and prognostic biomarkers for BC further research with a larger sample size is needed.

Survivin, encoded by BIRC5 gene (baculoviral IAP repeat containing 5), belongs to the family of inhibitors of apoptosis proteins (IAPs). In mammalian cells it participates in the control of mitosis, apoptosis regulation, and cellular stress response. Its expression is increased in almost all types of cancers. The aim of this study was to investigate the role of BIRC5 polymorphisms in breast cancer (BC) and to connect survivin expression with various clinicopathological characteristics of BC patients. Blood and archival tumour tissue samples were collected from 26 BC patients from Croatia. Survivin expression was determined immunohistochemically. BIRC5 promoter, coding region, and 3'UTR were genotyped. DNA from 74 healthy women was used as control. BIRC5 polymorphisms and survivin expression were tested against age of onset, histological grade, tumour type and size, lymph node status, oestrogen, progesterone, Her2, and Ki67 status. Numbers of samples with weak, moderate, and strong survivin expression were 9 (33.3%), 11 (40.7%), and 7 (25.9%), respectively. Most patients had nuclear survivin staining (92.6%). High survivin expression was significantly associated with negative oestrogen receptor status (p=0.007) and positive Ki67 expression (p=0.032). Ki67 expression was also positively correlated with histological grade (p=0.0009). Fourteen polymorphisms were found in BC samples, located mostly in promoter and 3'UTR of BIRC5. There was no significant difference in the distribution of polymorphisms between BC and control samples. Among clinicopathological characteristics of BC patients, alleles of five BIRC5 polymorphisms were associated with younger age of onset: c.-644T>C (55.8 years [y] vs. 48.1 y; p=0.006), c.-241C>T (54.2 y vs. 45.0; p=0.029), c.9809T>C (55.8 y vs. 48.1 y; p=0.006), c.-1547C>T (58.3 y vs. 50.9 y; p=0.011), and c.9386T>C (50.8 y vs. 59.5 y; p=0.004). To assess the significance of BIRC5 polymorphisms and survivin expression as predictive and prognostic biomarkers for BC further research with a larger sample size is needed.

Melanocortins, i.e., melanocyte stimulating hormones (MSH) are peptides with strong antiinflammatory effects. The most investigated aspects of γ2-MSH are related to cardiovascular effects and natriuresis, with limited research available about its anti-inflammatory and cytoprotective effects. The aims of this study were: 1) to examine the effects of γ2-MSH and its derivative [D-Trp(8)]-γ2-MSH on the acetaminophen model of liver damage in CBA mice; 2) to evaluate the modulation of γ2-MSH hepatoprotection by melanocortin subtypes 3 and 4 receptor antagonists SHU 9119 and HS 024; 3) to define the importance of central MSH pharmacophore region (HFRW) by using antisense peptides LVKAT and VKAT. In this study, specific antagonists and antisense peptides were used to target central pharmacophore region of γ2-MSH and [D-Trp(8)]-γ2-MSH, enabling the evaluation of hepatoprotection from the standpoint of the receptor and pharmacophore blockade. The criteria for monitoring the effects of the hormones on the liver damage were alanine transaminase, aspartate transaminase activities (U/L), and pathohistological scoring of liver necrosis (scale 0-5). γ2-MSH (0.24 mg/kg) indicated hepatoprotective effects in comparison to control (p < 0.001). In contrast, [D-Trp(8)]-γ2-MSH did not show any hepatoprotective effects. Application of antagonists SHU 9119 and HS 024, and antisense peptides LVKAT and VKAT, also did not show any hepatoprotective effects. In fact, when combined with γ2-MSH, it annulled its hepatoprotective effect. The results provide evidence for hepatoprotective and antiinflammatory effects of the γ2-MSH in the liver.

mutS homolog 2 (MSH2) deficiency may be involved in the development of microsatellite instability found in certain sporadic colorectal tumors. In addition to mutations or loss of heterozygosity resulting in complete loss of MSH2 function, polymorphisms affecting MSH2 expression have been also identified. Therefore, the aim of this study was to examine MSH2 status in sporadic colon cancer.MSH2 status was examined at the DNA, RNA and protein levels through loss of heterozygosity (LOH) analysis, quantitative real-time PCR and immunohistochemistry. MSH2 IVS10+12AG polymorphism was examined by real-time single nucleotide polymorphism genotyping.MSH2 LOH was more frequent in tumors larger than 5 cm (p=0.032), mRNA expression was also significantly lower and the same expression pattern was present in the corresponding normal mucosa of the same patient (p=0.013 and p=0.008, respectively). No association was found between IVS10+12AG polymorphism and susceptibility to sporadic colon cancer.Altered MSH2 expression detected in sporadic colon tumors pointing to its role in colorectal tumorigenesis without a hereditary component.

Background: mutS homolog 2 (MSH2) deficiency may be involved in the development of microsatellite instability found in certain sporadic colorectal tumors. In addition to mutations or loss of heterozygosity resulting in complete loss of MSH2 function, polymorphisms affecting MSH2 expression have been also identified. Therefore, the aim of this study was to examine MSH2 status in sporadic colon cancer. Materials and Methods: MSH2 status was examined at the DNA, RNA and protein levels through loss of heterozygosity (LOH) analysis, quantitative real-time PCR and immunohistochemistry. MSH2 IVS10+12A>G polymorphism was examined by real-time single nucleotide polymorphism genotyping. Results: MSH2 LOH was more frequent in tumors larger than 5 cm (p=0.032), mRNA expression was also significantly lower and the same expression pattern was present in the corresponding normal mucosa of the same patient (p=0.013 and p=0.008, respectively). No association was found between IVS10+12A>G polymorphism and susceptibility to sporadic colon cancer. Conclusion: Altered MSH2 expression detected in sporadic colon tumors pointing to its role in colorectal tumorigenesis without a hereditary component.

Cilj: Prikazati rezultate primjene kontrastnog ultrazvuka (engl. contrast-enhanced ultrasound – CEUS) u tercijarnom centru radi dijagnostičke karakterizacije jetrenih tumora. Bolesnici i metode: Retrospektivno su analizirani rezultati CEUS-a pri definiranju vrste jetrenih tumora u sukcesivno uključenoj kohorti bolesnika (od 1. siječnja 2015. do 31. prosinca 2016. godine). Definitivna dijagnoza vrste tumora postavljena je na temelju standardizirane kliničke obrade, što je uključivalo pregled magnetskom rezonancijom i/ili višeslojnim CT-om, tkivno uzorkovanje (biopsija ili citološka punkcija) u slučajevima kada nalaz nije bio jasan te patohistološku analizu reseciranog materijala operiranih bolesnika. Rezultati: U 90 bolesnika (48 M, 42 Ž, 57 ± 15 godina) analizirana su 132 jetrena tumora (68 benignih i 64 maligna), prosječne veličine 30 (8 – 130) mm. Nalaz CEUS-a imao je visoku točnost (senzitivnost 98,44%, specifičnost 83,82%, površina ispod krivulje ROC 0,91) za razlikovanje benignih tumora od malignih. Zaključak: Rezultati našeg Centra potvrđuju visoku dijagnostičku točnost i neškodljivost CEUS-a u karakterizaciji jetrenih tumora., Aim: To demonstrate the results of contrast-enhanced ultrasound (CEUS) for diagnostic characterization of liver tumors (LT) in a tertiary-care hospital. Patients and methods: Results of CEUS examinations of LT in successively included patients from January 1st 2015 to December 31st 2016 were retrospectively analyzed. Definite diagnosis was made through standardized clinical workup including MRI and/or MSCT, tissue sampling (biopsy or cytology) in cases when imaging was inconclusive, as well as pathohistological analysis of resected specimens in patients that underwent surgery. Results: In 90 patients (48 M, 42 F; age 57±15) 132 LT were analyzed (68 benign and 64 malignant), with average size of 30 (8-130) mm. CEUS demonstrated excellent diagnostic performance to differentiate between benign and malignant LT (sensitivity 98.44%, specifi city 83.82%, AUC 0.91), without adverse reactions. Conclusion: Our results demonstrate high diagnostic accuracy and safety of CEUS for characterization of LT.

The failure of the AJCC/UICC staging system in predicting prognosis in intermediate-stage of colorectal cancers (CRCs) may be overcome by adding tumor budding (TB) in pathology report as the presence of high-grade TB has been consistently associated with negative clinicopathologic parameters in gastrointestinal tumors, especially in CRCs. Over the past few decades, numerous methods of assessing TB in CRC have been proposed, with variations in the area of assessment, cut-off values, and use of standard hematoxylin and eosin (H&E) stained slides vs. cytokeratin immunohistochemistry. This review summarizes previous studies in this scientific area and resulting guidelines. The concept of whether peritumoral budding (PTB) vs. intratumoral budding (ITB), or both, should be assesed is still under discussion. The original studies on TB utilized PTB, or assesment of budding at the invasive front of the CRC, and current guidelines pertain to that. Budding category and tumor grade are not the same and TB has an independent prognostic value and should be taken into account along with other clinicopathological factors in a multidisciplinary setting. TB should be routinely reported in stage II CRC, next to other high-risk factors, in order to aid the decision for adjuvant therapy., Neuspjeh AJCC/UICC staging sustava u predviđanju prognoze u srednjem stadiju karcinoma debelog crijeva i rektuma (KDCR) može se prevladati dodavanjem tumorskog pupanja (TP) u nalaz patologa jer je prisutnost visokog gradusa TP dosljedno povezivana s negativnim klinikopatološkim parametrima u gastrointestinalnih tumora, posebno u KDCR. Tijekom proteklih nekoliko desetljeća predložene su brojne metode procjene TP u KDCR, s varijacijama područja procjene, graničnim vrijednostima i korištenjem standardnih hematoksilin-eozinom obojenih preparata nasuprot imunohistokemiji citokeratinom. Ovaj pregledni članak sažima dosadašnja istraživanja u ovom znanstvenom području i proizašle smjernice. Još uvijek se razmatra koncept treba li se procjenjivati peritumorsko pupanje (PTP) ili intratumorsko pupanje (ITP), ili oboje. Izvorne studije o TP-u koristile su PTP ili procjenu pupanja na invazivnom rubu KDCR, a trenutne se smjernice odnose na to. Kategorija pupanja i stupanj tumora nisu isti, a TP ima nezavisnu prognostičku vrijednost te ga treba uzeti u obzir zajedno s drugim kliničkopatološkim čimbenicima u multidisciplinarnom okruženju. TP bi trebalo rutinski izvijestiti u stadiju II KDCR, pored drugih visokorizičnih čimbenika, kako bi se pomogla odluka o adjuvantnoj terapiji.

The failure of the AJCC/UICC staging system in predicting prognosis in intermediate-stage of colorectal cancers (CRCs) may be overcome by adding tumor budding (TB) in pathology report as the presence of high-grade TB has been consistently associated with negative clinicopat- hologic parameters in gastrointestinal tumors, especially in CRCs. Over the past few decades, numerous methods of assessing TB in CRC have been proposed, with variations in the area of assessment, cut-off values, and use of standard hematoxylin and eosin (H&E) stained slides vs. cytokeratin immunohistochemistry. This review summarizes previous studies in this scientific area and resulting guidelines. The concept of whether peritumoral budding (PTB) vs. intratumo- ral budding (ITB), or both, should be assesed is still under discussion. The original studies on TB utilized PTB, or assesment of budding at the invasive front of the CRC, and current guidelines pertain to that. Budding category and tumor grade are not the same and TB has an independent prognostic value and should be taken into account along with other clinicopathological factors in a multidisciplinary setting. TB should be routinely reported in stage II CRC, next to other high-risk factors, in order to aid the decision for adjuvant therapy.

This study was aimed at the analysis of mononucleotide repeats -462T(15) and -4T(12) in the SMAD4 gene promoter in sporadic colon adenocarcinoma tissue of Croatian patients. The analysis has included 60 pairs of samples of colon tumor and adjacent normal tissue. The number of thymidines in the tracts -462T(15) and -4T(12) of the SMAD4 gene promoter was determined by PCR with fluorescently labeled primers followed by the analysis of obtained DNA fragments by capillary electrophoresis. In the normal colon tissue two haplotypes were present: -462T(15)/-4T(12) in 51 patients (85%) and -462T(16)/-4T(12) in 9 patients (15%). Among the cases with haplotype -462T(15)/-4T(12) detected in normal colon tissue, in 5 cases (8%) malignant tissue displayed different haplotypes: 462T(10)/-4T(10), -462T(12)/-4T(12), 462T(13)/-4T(11), -462T(14)/-4T(10) and -462T(15)/-4T(11). Haplotype -462T(14)/-4T(10) was previously found to be associated with significantly decreased SMAD4 gene promoter activity in comparison to the wild type, while the other detected haplotypes remain to be functionally characterized. This study has shown that functionally relevant somatic alterations of the SMAD4 gene promoter are found in some colon cancer tumors. Although not as frequent in colon as in pancreatic cancer, they may be of significance for certain cases and their role in colon tumorigenesis should be investigated further.

This review focuses on the described effects of BPC 157 on blood vessels after different types of damage, and elucidate by investigating different aspects of vascular response to injury (endothelium damage, clotting, thrombosis, vasoconstriction, vasodilatation, vasculoneogenesis and edema formation) especially in connection to the healing processes. In this respect, BPC 157 was concluded to be the most potent angiomodulatory agent, acting through different vasoactive pathways and systems (e.g. NO, VEGF, FAK) and leading to optimization of the vascular response followed, as it has to be expected, by optimization of the healing process. Formation of new blood vessels involves two main, partly overlapping mechanisms, angiogenesis and vasculogenesis. The additional mechanism of arteriogenesis is involved in the formation of collaterals. In conjunction with blood vessel function, we at least have to consider leakage of fluid/proteins/plasma, resulting in edema/exudate formation as well as thrombogenesis. Blood vessels are also strongly involved in tumor biology. In this aspect, we have neoangiogenesis resulting in pathological vascularization, vascular invasion resulting in release of metastatic cells and the phenomenon of homing resulting in formation of secondary tumors – metastases.

Objective To report two cases of Merkel cell carcinoma (MCC) metastatic to the pancreas diagnosed with endoscopic ultrasound-guided-fine needle aspiration (EUS-FNA) and to add the case of concomitant chronic lymphocytic leukaemia/small lymphocytic lymphoma (CLL/SLL) and MCC to the literature. The aim is to alert the cytopathologists once more to the problems of differential diagnosis of pancreatic metastasis of MCC and to describe the possibilities of ancillary methods performed on direct cytological smears. Methods EUS-FNA procedures were performed according to standard institution protocol, using 22-G needles with cytopathologist on-site. Based on rapid on-site evaluation (ROSE), additional passes were made for immunocytochemistry (ICC). A mini panel of antibodies was used to aid the differential diagnosis. Results Smears revealed a dispersed pattern of small round cells with scant cytoplasm, round nuclei with inconspicuous nucleoli and occasional nuclear moulding, suspicious of small cell carcinoma. Results of ICC applied to the direct cytological smears were as follows: LCA negative, Cytokeratin (clone MNF116) positive, TTF-1 negative, CD 56 positive, NSE weakly positive, Chromogranin A weakly positive and CK20 positive, in one case in a dot-like perinuclear pattern. The diagnosis of MCC was made. Conclusion Increasing incidence of MCC warrants the inclusion of MCC in the differential diagnosis of tumours of small round blue cell morphology even in unusual sites. The cytomorphological features coupled with an ICC panel are usually enough to make a confident diagnosis of MCC. EUS-FNA is a minimally invasive technique which enables sampling adequate tissue for all the ancillary methods eventually needed to support the diagnosis.

Primary myelofibrosis (PMF) is a neoplastic disease originating from clonally transformed pluripotent hematopoietic stem cell. Heat shock proteins (HSP) are expressed by cells in response to different stress stimuli, thus allowing cells to survive in otherwise lethal conditions. Heat shock protein 27 (HSP27) is a small HSP transcribed from heat shock protein family B member 1 (HSPB1) gene. Increased HSP27 expression and phosphorylation were observed in a large number of neoplastic diseases and they have mostly been associated with more aggressive disease features, metastatic behavior, resistance to chemotherapy and poor prognosis. However, these findings are not universal for different malignant diseases and higher HSP27 expression was recognized as a positive marker in some situations. Increased HSP27 expression was also shown to be atheroprotective. In this study, we show the genetic upregulation of HSPB1/HSP27 in myelofibrosis and its potentially favorable impact on survival.

Low-contrast images, such as color microscopic images of unstained histological specimens, are composed of objects with highly correlated spectral profiles. Such images are very hard to segment. Here, we present a method that nonlinearly maps low-contrast color image into an image with an increased number of non-physical channels and a decreased correlation between spectral profiles. The method is a proof-of-concept validated on the unsupervised segmentation of color images of unstained specimens, in which case the tissue components appear colorless when viewed under the light microscope. Specimens of human hepatocellular carcinoma, human liver with metastasis from colon and gastric cancer and mouse fatty liver were used for validation. The average correlation between the spectral profiles of the tissue components was greater than 0.9985 and the worst case correlation was greater than 0.9997. The proposed method can potentially be applied to the segmentation of low-contrast multichannel images with high spatial resolution that arise in other imaging modalities.

We propose an offset-sparsity decomposition (OSD) method for the enhancement of a color microscopic image of a stained specimen. The method decomposes vectorized spectral images into offset terms and sparse terms. A sparse term represents an enhanced image, and an offset term represents a “shadow.” The related optimization problem is solved by computational improvement of the accelerated proximal gradient method used initially to solve the related rank-sparsity decomposition problem. Removal of an image-adapted color offset yields an enhanced image with improved colorimetric differences among the histological structures. This is verified by a no-reference colorfulness measure estimated from 35 specimens of the human liver and 1 specimen of the mouse liver stained with hematoxylin and eosin, 6 specimens of the mouse liver stained with Sudan III, and 3 specimens of the human liver stained with the anti-CD34 monoclonal antibody. The colorimetric difference improves on average by 43.86% with a 99% confidence interval (CI) of [35.35%, 51.62%]. Furthermore, according to the mean opinion score, estimated on the basis of the evaluations of five pathologists, images enhanced by the proposed method exhibit an average quality improvement of 16.60% with a 99% CI of [10.46%, 22.73%].

Melanocortins, i.e., melanocyte stimulating hormones (MSH) are peptides with strong antiinflammatory effects. The most investigated aspects of γ2-MSH are related to cardiovascular effects and natriuresis, with limited research available about its anti-inflammatory and cytoprotective effects. The aims of this study were: 1) to examine the effects of γ2-MSH and its derivative [D-Trp(8)]-γ2-MSH on the acetaminophen model of liver damage in CBA mice; 2) to evaluate the modulation of γ2-MSH hepatoprotection by melanocortin subtypes 3 and 4 receptor antagonists SHU 9119 and HS 024; 3) to define the importance of central MSH pharmacophore region (HFRW) by using antisense peptides LVKAT and VKAT. In this study, specific antagonists and antisense peptides were used to target central pharmacophore region of γ2-MSH and [D-Trp(8)]-γ2-MSH, enabling the evaluation of hepatoprotection from the standpoint of the receptor and pharmacophore blockade. The criteria for monitoring the effects of the hormones on the liver damage were alanine transaminase, aspartate transaminase activities (U/L), and pathohistological scoring of liver necrosis (scale 0-5). γ2-MSH (0.24 mg/kg) indicated hepatoprotective effects in comparison to control (p0.001). In contrast, [D-Trp(8)]-γ2-MSH did not show any hepatoprotective effects. Application of antagonists SHU 9119 and HS 024, and antisense peptides LVKAT and VKAT, also did not show any hepatoprotective effects. In fact, when combined with γ2-MSH, it annulled its hepatoprotective effect. The results provide evidence for hepatoprotective and antiinflammatory effects of the γ2-MSH in the liver.

The effects of the gastric pentadecapeptide BPC 157, administered topically or systemically in burned mice were investigated. This agent is known to have a beneficial effect in variety models of gastrointestinal lesions, as well as on wound or fracture healing, also when given locally. Deep partial skin thickness burns (1.5cmx1.5cm) covering 20% of total body area, were induced under anesthesia on the back of the mice by controlled burning and gastric lesions were assessed 1, 2, 3, 7, 14 and 21 days following injury. The first application was immediately following burning, and thereafter, once daily, last 24h before sacrifice. In the initial experiments, exposition to direct flame for 5 seconds, the BPC 157 was applied at 10 mg or 10 ng/kg b.w. i.p. by injection or alternatively, topically, at the burn, as a thin layer of cream (50 mg of BPC 157 dissolved in 2 ml of distilled water was mixed with 50 g of commercial neutral cream (used also as local vehicle-control)), while silver sulfadiazine 1% cream was a standard agent acting locally. Others received no local medication: they were treated i.p. by injection of distilled water (distilled water-control) or left without any medication (control). In subsequent experiments involving deeper burns (direct flame for 7 seconds), BPC 157 creams (50 ľg, 5 ľg, 500 ng, 50 ng or 5 ng of BPC 157 dissolved in 2 ml of distilled water was mixed with 50 g of commercial neutral cream), or vehicle as a thin layer of cream, were applied topically, at the burn. Compared with untreated controls, in both experiments, in the BPC 157 cream treated mice all parameters of burn healing were improved throughout the experiment: less edema was observed and inflammatory cells number was decreased, less necrosis, an increased number of capillaries along with an advanced formation of dermal reticulin and collagen fibers and an increased number of preserved follicles were observed. Two weeks after injury, BPC 157 cream-treated mice completely reversed the otherwise poor reepithelization ratio noted in the untreated control or mice treated with only cream vehicle. Tensiometry investigation showed an increased breaking strength and relative elongation of burned skin, while water content in burned skin decreased. This was, however, not the case with the vehicle cream or silver sulfadiazine cream-mice. Relative to the control values, in silver sulfadiazine cream-treated mice, only collagen fiber formation was increased, in addition to a decreased inflammatory cells number. Relative to control values, BPC 157 given intraperitoneally decreased the number of inflammatory cells, lowered water content in burned skin, and raised breaking strength and relative elongation of burned skin during tensiometry. Through the experimental period gastric lesions were continously noted in all thermally injured mice left without local medication and they were consistently attenuated only by BPC 157 treatments: either given intraperitoneally (at either dose), or given locally (at either concentrations). Other treatments (i.e., local treatment with silver sulfadiazine cream or neutral cream in mice subjected for 5 seconds to direct flame), lead to only poor, if any attenuation. This stable gastric pentadecapeptide appears to be active and give a stimulation to burn healing at the defect site. The agent may act by causing an upregulation of the growth factors, as well as influences other local factors.

Recently, we showed cysteamine-duodenal lesions without gastric acid, since they were induced also in gastrectomized rats, as in naive rats, and they were inhibited by the novel stomach pentadecapeptide BPC 157 as well as standard antiulcer drugs (i.e. cimetidine, ranitidine, omeprazole, bromocriptine, atropine). Therefore, as an advantage of considering cysteamine as a directly acting cytotoxic agent and mentioned agents as direct cytoprotective agents, the present focus was on the ulcerogenic effect of cysteamine and protective effect of gastroduodenal antiulcer agents outside upper gastrointestinal tract (i.e. in colon). Intrarectal administration of the cysteamine (200 or 400 mg/kg b.w) produced severe colon lesions (i.e. transmural inflammation with serosal involvement) in rats (30 min–72h-experimental period), apparently distinctive from smaller lesions after non-specific irritant enema [diluted HCl solution, pH 3.8 (adjusted to pH of cysteamine solution (pH 3.8)]. All of the tested antiulcer agents were applied simultaneously with cysteamine enema (8 cm from the anus, in a volume of the 1.0 ml/rat) intraperitoneally (i.p.), intragastrically (i.g.) or intrarectally (i.r.). Pentadecapeptide BPC 157 (10 μg or 10 ng/kg b.w.), given in either regimen, previously shown to have, besides others, a particular beneficial activity just in the intestinal mucosa, inhibited these cysteamine colon lesions (assessed after 30 min, 60 min, 180 min, 24 h, 48 h, 72 h following cysteamine in a dose of either 200 or 400 mg/kg i.r.). Cysteamine-colon lesions were also attenuated by standard antiulcer agents (mg/kg b.w.), given i.p., i.g., or i.r., such as ranitidine (10), cimetidine (50), omeprazole (10), atropine (10), together with methylprednisolone (1), and sulphasalazine (50, i.r.), assessed 30 min following application of 200 mg of cysteamine. Finally, standard cysteamine duodenal lesions (assessed 24 h after a subcutaneous application of 400 mg/kg of cysteamine) were also attenuated by these agents application (given in the same doses, i.p., 1 h before cysteamine), with only exception to sulphasalazine. Thus, the extended cysteamine specific ulcerogenic effect, cysteamine colon/duodenum lesion-link and an extenuation of agents protection from upper to lower part of gastrointestinal tract (i.e. stomach pentadecapeptide BPC 157, standard antiulcer agents, cimetidine, ranitidine, atropine, omeprazole) and vice versa (remedies for inflammatory bowel disease) evidenced in the present study may be potentially important for both further experimental and clinical research.

Unlike severe gastric damage acutely induced by ethanol administration in rat, the ulcerogenic effect of chronic alcohol administration (3.03 g/kg b.w. or 7.28 g/kg b.w.) given in drinking water, producing liver lesions and portal hypertension, is far less investigated. Therefore, focus was on the antiulcer effect of the gastric pentadecapeptide BPC 157, GEPPPGKPADDAGLV, M.W. 1419, known to have a beneficial effect in variety of gastrointestinal lesions models (10 microg or 10 ng/kg b.w. i.p. or i.g.), ranitidine (10 mg/kg b.w. i.g.) and propranol (10 mg/kg b.w. i.g.) or saline (5 ml/kg b.w. i.p./i.g.; control). They were given once daily (1) throughout 10 days preceding alcohol consumption, (2) since beginning of alcohol drinking till the end of the study, (3) throughout the last month of alcohol consumption, 2 months after alcohol drinking had been initiated. Gastric lesions were assessed, at the end of 3 months drinking [(1), (2)] or with respect to therapeutic effect of medication before medication or at the end of therapy. Pentadecapeptide BPC 157, ranitidine and propranolol may prevent gastric lesion development if given prophylactically, before alcohol drinking. Likewise, they attenuate the lesion appearance given once daily throughout the drinking period. Importantly, when given therapeutically, they may antagonize otherwise pertinent lesion presence in stomach mucosa of the drinking rats. Thus, these results demonstrate that pentadecapeptide BPC 157, ranitidine and propranol may prevent, attenuate or reverse the gastric lesions appearance in chronically alcohol drinking rats, and may be used for further therapy, while the other studies showed that their effect (except to ranitidine) is parallel with their beneficial effect on liver lesion and portal hypertension.

Up to now, for gastric lesions potentiation or induction, as well as determination of endogenous dopamine significance, dopamine antagonist or dopamine vesicle depletor were given separately. Therefore, without combination studies, the evidence for dopamine significance remains split on either blockade of dopamine post-synaptic receptor or inhibition of dopamine storage, essentially contrasting with endogenous circumstances, where both functions could be simultaneously disturbed. For this purpose, a co-administration of reserpine and haloperidol, a dopamine granule depletor combined with a dopamine antagonist with pronounced ulcerogenic effect, was tested, and the rats were sacrificed 24 h after injurious agent(s) administration. Haloperidol (5 mg x kg(-1) b.w. i.p.), given alone, produced the lesions in all rats. Reserpine (5 mg x kg(-1) b.w. i.p.), given separately, also produced lesions. When these agents were given together, the lesions were apparently larger than in the groups injured with separate administration of either haloperidol or reserpine alone. Along with our previous results, when beneficial agents were co-administered, all dopaminomimetics (bromocriptine 10 mg, apomophine 1 mg, amphetamine 20 mg x kg(-1) i.p.) apparently attenuated the otherwise consistent haloperidol-gastric lesions. Likewise, an apparent inhibition of the reserpine-lesions was noted as well. However, if they were given in rats injured with combination of haloperidol and reserpine, their otherwise prominent beneficial effects were absent. Ranitidine (10 mg), omeprazole (10 mg), atropine (10 mg), pentadecapeptide BPC 157 (Gly-Glu-Pro-Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp-Ala-Gly-Leu-Val) (10 microg or 10 ng x kg(-1) i.p.) evidently prevented both haloperidol-gastric lesions and reserpine-gastric lesions. Confronted with potentiated lesions following a combination of haloperidol and reserpine, these agents maintained their beneficial effects, noted in the rats treated with either haloperidol or reserpine alone. The failure of dopaminomimetics could be most likely due to more extensive inhibition of endogenous dopamine system activity, and need for remained endogenous dopamine for their salutary effect, whereas the beneficial activities of ranitidine, omeprazole, atropine, pentadecapeptide BPC 157 following dopamine system inhibition by haloperidol+reserpine suggest their corresponding systems parallel those of dopamine system, and they may function despite extensive inhibition of endogenous dopamine system activity.

Recently, the effectiveness of pentadecapeptide BPC 157 and other anti-ulcer agents, called 'direct cytoprotection', was evidenced in totally gastrectomized rats duodenum challenged with cysteamine 24 h after surgery, and sacrificed 24 h after ulcerogen application. The further focus was on the possibility that this effect could be seen over a more prolonged period (1, 2, 4 weeks), and in other parts of the gastrointestinal tract (i.e. oesophagus). After the removal of the stomach, the oesophagus and jejunum were joined by a termino-lateral anastomosis. The animals were euthanized 7, 14 or 28 d after surgery, when oesophagitis was blindly assessed both macroscopically (percentage of ulcerations areas) and microscopically (percentage of areas of ulcers, regeneration and hyperplasia; number of inflammatory cells - polymorphonuclear and mononuclear). Starting 24 h after surgery, the medication was continuously given in the drinking water, in a volume of 12.5 mL/rat daily, until euthanasia at the end of the observation period, i.e. 7, 14, 28 d following surgery. Based on previous experiments, the doses of agents were daily calculated per kg b.w. as follows: BPC 157 125 mg or 125 ng, cholestyramine 2.5 mg, ranitidine 125 mg, sucralfate 725 mg, whereas controls received 72.5 mL x kg(-1) water. In support of these initial findings, and considering gastrectomized acid-free rats as an ideal model for long-term cytoprotective studies as well, pentadecapeptide BPC 157 markedly attenuated termino-lateral oesophagojejunal anastomosis-reflux oesophagitis also over a quite prolonged period. This efficacy was only partly shared by other anti-ulcer agents. After 1-week-old oesophagitis (microscopical assessment), but not after 2 or 4 weeks, less damaged mucosa was noted in rats drinking ranitidine or sucralfate compared to controls. Similar effectiveness was noted for cholestyramine. The obtained results were supported also by inflammatory cell assessment. Compared with control values, BPC 157-treated groups consistently presented less polymorphonuclears and less mononuclears in all assessed periods. Interestingly, the values obtained in other treated groups showed no difference compared with control values. Thus, despite limitations, a generalization supporting a direct importance of a common cytoprotective approach, could be clearly provided. A useful, long-lasting cytoprotective activity (apparently more prominent in BPC 157 rats, than in reference agents, ranitidine, sucralfate, as well as cholestyramine) may be a likely suitable therapy in otherwise resistant reflux oesophagitis conditions.

Purpose Inherited polymorphisms in immunomodulatory genes such as cytokines may contribute to variation in immunological response and genetic susceptibility for complex diseases, including cancer. TNFα can mediate tumor progression by inducing proliferation, invasion and metastasis of tumor cells. The aim of our study was to examine the allelic frequencies of TNFα promoter SNPs, − 1031 T/C, − 857 C/T, − 308 G/A and − 238 G/A, in patients with sporadic colon adenocarcinoma in order to investigate the possible role of these SNPs in susceptibility to sporadic colon cancer. Another aim of this study was to examine the influence of TNFα SNPs on TNFα mRNA and protein expression in colon tumors and their possible role in the development and progression of this type of tumor. Results The distribution of all four TNFα SNP genotypes in patients showed no significant difference compared to controls. No statistically significant difference in TNFα mRNA expression in tumors and corresponding normal mucous tissue was found (p = 0.14). A statistically significant (p = 0.028) difference was found in TNFα mRNA expression between histological grade 1 and histological grade 2 and 3 tumors. Additionally, a statistically significant correlation (p = 0.03) was found between TNFα-857 C/T genotypes and TNFα mRNA expression in tumor tissue. TNFα mRNA expression was significantly higher in the tumor tissue of patients with -857 CT and -857 TT genotypes. Most of the tumors (78.26%) were positive for TNFα protein. No correlation was found between the TNFα protein expression and clinicopathological characteristics as well as TNFα genotypes. However, patients with TNFα protein negative tumors had longer survival but the result was not statistically significant (p = 0.365). Conclusion Our results suggest the role of TNFα as one of the immunomodulatory genes in the progression of sporadic colon cancer.

NF2/Merlin was first identified through its association with neurofibromatosis type 2 (NF2). However, accumulating evidence suggests a more general involvement in tumorigenesis and, in particular, a broader role in tumor suppression. The aim of this study was to examine NF2/Merlin involvement in sporadic colorectal cancer. This study is the first to examine the role of NF2/Merlin in sporadic colorectal cancer through LOH analysis at the NF2 locus and mRNA expression analysis via quantitative RT-PCR of total NF2, NF2 isoform I and II. In addition, Merlin protein expression was assessed by immunohistochemistry and Western blotting. NF2 LOH was detected in 20.0 % of heterozygous cases and was found to be more frequent in tumors larger than 5 cm in diameter (p = 0.041) and in tumors with a less differentiated phenotype (p = 0.027). No differences were observed in total NF2 and NF2 isoform I/isoform II mRNA expression between the tumors and their corresponding normal mucous tissues. NF2 isoform II was the most predominant isoform in all samples analyzed. mRNA expression levels of total NF2 and isoforms I and II were significantly lower in poorly differentiated tumors (p = 0.033, p = 0.036 and p = 0.044, respectively). Weak Merlin immunostaining was more frequent in poorly differentiated tumors (p = 0.034) and tumors classified as Dukes’ C (p = 0.023). A distinct pattern of Merin phosphorylation was observed in tumors compared to normal mucous tissues. Our data indicate that NF2/Merlin may serve as a potential target in the management of colorectal cancer.