Correction: Warzecha, Z. et al. Protective Effect of Pretreatment with Acenocoumarol in Cerulein-Induced Acute Pancreatitis. Int. J. Mol. Sci. 2016, 17, 1709.

We would like to submit the correction to our published paper [...].

Correction: Warzecha, Z., et al. Therapeutic Effect of Low Doses of Acenocoumarol in the Course of Ischemia/Reperfusion-Induced Acute Pancreatitis in Rats. Int. J. Mol. Sci. 2017, 18, 882.

We would like to submit this correction to our published paper [...].

Rationale and feasibility of mucin expression profiling by qRT-PCR as diagnostic biomarkers in cytology specimens of pancreatic cancer.

BACKGROUND: Aberrantly expressed mucin glycoproteins (MUC) play important roles in pancreatic ductal adenocarcinoma (PDAC), yet their use as a diagnostic aid in fine-needle aspiration biopsy (FNAB) is poorly documented. The aim of this study was to investigate the rationale and feasibility of mucin (MUC1, MUC2, MUC3, MUC4, MUC5AC, and MUC6) expression profiling by RT-PCR for diagnostic applications in cytology. METHODS: Mucin expression was examined by RT-PCR and immunohistochemistry in specimens resected from patients with pancreatic (n = 101), ampullary (n = 23), and common bile duct (n = 10) cancers and 33 with chronic pancreatitis. Furthermore, mucin profiling by RT-PCR was prospectively compared in surgical and biopsy specimens of 40 patients with pancreatic solid tumours qualified for FNAB prior to surgery. RESULTS: A logistic regression model to distinguish PDAC from chronic pancreatitis using RT-PCR profiling included MUC3, MUC5AC, and MUC6. The same set of mucins differentiated ampullary and bile duct cancers from chronic pancreatitis. AUCs for the ROC curves derived from the two models were 0.95 (95%CI 0.87-0.99) and 0.92 (95%CI 0.81-0.98), respectively. The corresponding positive likelihood ratios were 6.02 and 5.97, while the negative likelihood ratios were 0.10 and 0.12. AUCs of ROC curves obtained by RT-PCR and immunohistochemistry demonstrated that both analytical methods were comparable. Surgical and cytological samples showed significantly correlated values of ΔCt for individual mucins with the overall Pearson's correlation coefficient r = 0.841 (P = 0.001). CONCLUSIONS: Mucin expression profiling of pancreatic cancer with RT-PCR is feasible and may be a valuable help in discriminating malignant lesions from chronic pancreatitis in FNAB cytology.

Therapeutic Effect of Low Doses of Acenocoumarol in the Course of Ischemia/Reperfusion-Induced Acute Pancreatitis in Rats.

Intravascular activation of coagulation is observed in acute pancreatitis and is related to the severity of this inflammation. The aim of our study was to evaluate the impact of acenocoumarol therapy on the course of acute pancreatitis induced in male rats by pancreatic ischemia followed by reperfusion. Acenocoumarol at a dose of 50, 100, or 150 µg/kg/dose was administered intragastrically once a day, starting the first dose 24 h after the initiation of pancreatic reperfusion. RESULTS: Histological examination showed that treatment with acenocoumarol reduces pancreatic edema, necrosis, and hemorrhages in rats with pancreatitis. Moreover, the administration of acenocoumarol decreased pancreatic inflammatory infiltration and vacuolization of pancreatic acinar cells. These findings were accompanied with a reduction in the serum activity of lipase and amylase, concentration of interleukin-1ß, and plasma d-Dimer concentration. Moreover, the administration of acenocoumarol improved pancreatic blood flow and pancreatic DNA synthesis. Acenocoumarol given at a dose of 150 µg/kg/dose was the most effective in the treatment of early phase acute pancreatitis. However later, acenocoumarol given at the highest dose failed to exhibit any therapeutic effect; whereas lower doses of acenocoumarol were still effective in the treatment of acute pancreatitis. CONCLUSION: Treatment with acenocoumarol accelerates the recovery of ischemia/reperfusion-induced acute pancreatitis in rats.

Semiquantitative immunohistochemistry for mucin (MUC1, MUC2, MUC3, MUC4, MUC5AC, and MUC6) profiling of pancreatic ductal cell adenocarcinoma improves diagnostic and prognostic performance.

AIMS: Mucin (MUC) glycoproteins are involved in various steps of the carcinogenesis and progression of human malignancies. The aim of this study was to verify whether semiquantitative evaluation of MUC staining by immunohistochemistry may help to differentiate pancreatic ductal cell adenocarcinoma (PDAC) from chronic pancreatitis and normal pancreas. METHODS AND RESULTS: Mucin expression was examined by immunohistochemistry in surgical specimens resected from 101 patients with PDAC and 33 with chronic pancreatitis, and in 40 normal pancreatic tissue specimens. A quickscore (QS, range 0-300) was calculated by multiplying staining intensity by the percentage of positive cells. A diagnostic model was developed for MUC QS (MUC1, MUC2, MUC3, MUC4, MUC5AC, and MUC6), based on a receiver operating characteristic (ROC) curve and logistic regression analysis. Median QS values for MUC1 and MUC5AC were significantly higher for PDAC, whereas patients with non-malignant tissues had higher values for MUC3 and MUC6. The area under the curve for the ROC curve derived from the diagnostic model including MUC3, MUC5AC and MUC6 was 0.96 [95% confidence interval (CI) 0.91-0.98], with 85% sensitivity and 94% specificity. Median QS values for MUC2 were significantly higher in patients with less advanced tumours, whereas venous invasion was associated with a lower QS for MUC6. Moreover, multivariate survival analysis revealed that low MUC6 expression was a negative prognostic factor, with a hazard ratio of 1.73 (95% CI 1.07-2.81). CONCLUSIONS: The three-MUC diagnostic model (MUC3, MUC5AC, and MUC6) showed an excellent ability to discriminate pancreatic cancer from non-malignant tissues, and yielded information that may prove useful for the development of clinical applications.

IgG4-related disease in the head and neck region: report of two cases and review of the literature.

IgG4-related disease (IgG4-RD) is a rare immune-mediated condition characterized by extensive tissue fibrosis and infiltration by immunoglobulin G4 positive plasma cells in a single organ or systemic appearance. Two cases are presented including an unusual case of a 30-year-old man with IgG4-RD appearing simultaneously in the cervical lymph nodes, ethmoid, maxillary sinuses, and upper gingiva, with spontaneous loss of teeth. According to the literature, this is the first case with loss of teeth occurring in the course of the disease. The second case is a 46-year-old man suffering from IgG4-related chronic sclerosing sialadenitis of the right submandibular gland.

Protective Effect of Pretreatment with Acenocoumarol in Cerulein-Induced Acute Pancreatitis.

Coagulation is recognized as a key player in inflammatory and autoimmune diseases. The aim of the current research was to examine the effect of pretreatment with acenocoumarol on the development of acute pancreatitis (AP) evoked by cerulein. METHODS: AP was induced in rats by cerulein administered intraperitoneally. Acenocoumarol (50, 100 or 150 µg/kg/dose/day) or saline were given once daily for seven days before AP induction. RESULTS: In rats with AP, pretreatment with acenocoumarol administered at the dose of 50 or 100 µg/kg/dose/day improved pancreatic histology, reducing the degree of edema and inflammatory infiltration, and vacuolization of acinar cells. Moreover, pretreatment with acenocoumarol given at the dose of 50 or 100 µg/kg/dose/day reduced the AP-evoked increase in pancreatic weight, serum activity of amylase and lipase, and serum concentration of pro-inflammatory interleukin-1ß, as well as ameliorated pancreatic DNA synthesis and pancreatic blood flow. In contrast, acenocoumarol given at the dose of 150 µg/kg/dose did not exhibit any protective effect against cerulein-induced pancreatitis. CONCLUSION: Low doses of acenocoumarol, given before induction of AP by cerulein, inhibit the development of that inflammation.

Age- and degeneration-related variations in cell density and glycosaminoglycan content in the human cervical intervertebral disc and its endplates.

The first aim of this study was to quantify cell density in cervical intervertebral discs (IVDs) and endplates of varying age and degeneration grade. The second aim was to analyze glycosaminoglycan (GAG) content in cervical IVDs and their endplates. Sixty cervical IVDs were excised from 30 human cadavers, not later than 24 hours post-mortem. Each sample underwent sectioning. Half of each sample underwent GAG content analysis using the dimethylmethylene blue binding assay. The other half underwent histological processing, histological degeneration grading, and cell density assessment using the Abercrombie method. The nucleus pulposus (NP) (4218 ± 417 cells/mm³) had significantly higher cell density than the anterior annulus fibrosus (AF) (3283 ± 438 cells/mm³; p < 0.0001), and similar cell density (4464 ± 551 cells/mm³; p = 0.36) to the posterior AF. Cell density was similar throughout the different regions of the endplate. The NP (619 ± 178 µg/mg dry weight) had a significantly higher GAG content than both the anterior (428 ± 199 µg/mg dry weight; p < 0.0001) and posterior AF (524 ± 218 µg/mg dry weight; p < 0.0001). In conclusion, this study introduces detailed 3D maps of cervical IVD and endplate cell density and GAG content. Furthermore, it shows that cervical IVDs and their endplates only slightly differ, in terms of cell density and GAG content, from lumbar IVDs.

Degeneration and calcification of the cervical endplate is connected with decreased expression of ANK, ENPP-1, OPN and TGF-ß1 in the intervertebral disc.

The aim of this study was to clarify the relationship between the expression of ALP, ANK, ENPP-1, OPN and TGF-ß1 in the intervertebral disc (IVD), and cervical vertebral endplate calcification and degeneration. Sixty cervical IVDs were excised from 30 human cadavers. Each cadaver was assessed macroscopically for degeneration (Thompson's classification), and then underwent histological processing, regular staining (hematoxylin and eosin, Masson-Goldner trichrome and alcian blue-PAS), immunohistochemistry (ALP, ANK, ENPP-1, OPN and TGF-ß1), microscopic degeneration grading (Boos classification), and assessment of endplate calcification. The mean age ± SD of the cadavers was 51.4 ±19.5. The percentage of endplate calcification significantly correlated with the degree of endplate and IVD degeneration graded using Boos's score (both r = 0.91; p < 0.0001). The intensity and number of stained cells per FOV markedly decreased, for ANK, ENPP-1, and TGF-ß1, with the grade of IVD degeneration, regardless of the analyzed IVD region. This was not true only for ALP, which demonstrated an increasing trend corresponding to the degree of IVD degeneration. The expression of OPN was low throughout all analyzed regions, regardless of the degree of degeneration. Modulating the expression of the abovementioned proteins, especially ANK and TGF-ß1, may be a new way to prevent degeneration and calcification of the IVD.

Analysis of metallothionein and vimentin immunoreactivity in pharyngeal squamous cell carcinoma and its microenvironment.

Metallothionein (MT) has been shown to have pro-proliferative anti-apoptotic activity and to be involved in microenvironment remodeling. The aim of this study has been to determine whether the changes in MT and vimentin immunoreactivity observed in cancer and its microenvironment are related to the local spread of the disease. The immunoreactivity levels of both MT and vimentin were evaluated together with CD56 and CD57 antigens in 49 tissue samples taken from patients with squamous cell carcinoma originating from the palatine tonsils and in 20 tissue samples derived from patients with chronic tonsillitis (the reference group). MT immunoreactivity levels were statistically significantly higher in the tissue samples from squamous cell carcinoma than in those of the reference group and also higher in the squamous cell carcinoma samples compared with the stromal samples. Moreover, stromal fibroblasts exhibited high vimentin and MT immunoreactivity levels. Statistically significantly higher MT immunoreactivity levels within the tumor cells were identified in patients with the presence of lymph node metastases in contrast to those patients without such metastases. Vimentin was detected in both the tumor and the stromal tissue samples and presented an interesting pattern of staining strongly expressed within the stroma and the septal architecture of the tumor. The number of CD56- and CD57-positive lymphocytes identified in tissue samples both from squamous cell carcinoma and from the stroma was statistically significantly lower than that in the reference group. MT expression by tumor cells is thus associated with an aggressive phenotype of the tumor and the ability to create metastases.

Malignant transformation in the course of a dentigerous cyst: a problem for a clinician and a pathologist. Considerations based on a case report.

Primary intraosseous squamous cell carcinoma (PIOSCC) is a rarely reported neoplasm resulting from malignant transformation of a dentigerous cyst of the mandible or maxilla. Until 2010, only 116 cases had been described. The diagnosis of PIOSCC is difficult because of non-specific symptoms. A case of a 66-year-old patient with PIOSCC arising from a dentigerous cyst of the mandible is presented. Both pathologists and clinicians should be aware of the probability of malignant transformation of dentigerous cysts during the two-stage treatment. The patient should be subject to regular clinical and radiographic examination.

Increased nitric oxide availability attenuates high fat diet metabolic alterations and gene expression associated with insulin resistance.

BACKGROUND: High fat diet impairs nitric oxide (NO) bioavailability, and induces insulin resistance. The link between NO availability and the metabolic adaptation to a high fat diet is not well characterized. The purpose of this study was to investigate the effect of high fat diet on metabolism in mice with decreased (eNOS-/-) and increased (DDAH overexpressed) NO bioavailability. METHODS: eNOS-/- (n = 16), DDAH (n = 24), and WT (n = 19) mice were fed a high fat diet (HFD) for 13 weeks. Body weight, biochemical parameters, adipokines and insulin were monitored. The matrigel in vivo model with CD31 immunostaining was used to assess angiogenesis. Gene expression in adipose tissues was analyzed by microarray and Real Time PCR. Comparisons of the mean values were made using the unpaired Student t test and p < 0.05 were considered statistically significant. RESULTS: eNOS-/- mice gained less weight than control WT and DDAH mice. In DDAH mice, a greater increase in serum adiponectin and a lesser increment in glucose level was observed. Fasting insulin and cholesterol levels remained unchanged. The angiogenic response was increased in DDAH mice. In adipose tissue of DDAH mice, genes characteristic of differentiated adipocytes were down-regulated, whereas in eNOS-/- mice, genes associated with adipogenesis, fatty acid and triglyceride synthesis were upregulated. CONCLUSIONS: Our results indicate that increased NO availability attenuates some HFD induced alterations in metabolism and gene expression associated with insulin resistance.

A rare case of aggressive, hereditary paraganglioma associated with a pathogenic variant in SDHD.

Not required for Clinical Vignette.

Combination of endosonography-guided fine-needle aspiration and conventional endoscopic techniques in sarcoidosis diagnosis. Optimal strategy to achieve high diagnostic yield.

INTRODUCTION: Diagnosis of sarcoidosis is based on clinical status and radiologic specific findings. Tissue confirmation of noncaseating granulomas is crucial. Pathological confirmation of pulmonary sarcoidosis is most commonly accomplished by bronchoscopy, which has a diagnostic yield of approximately 60%-70%. OBJECTIVES: In this prospective study, we analysed potential benefit of EBUS-TBNA and EBB combination, application of cell blocks and smears with puncturing more than one station of lymph nodes in order to determine optimal strategy in diagnosis of sarcoidosis. METHODS: About 133 patients with suspicion of sarcoidosis (stage I and stage II) were included in this study. Each patient underwent conventional bronchoscopy with endobronchial biopsy (EBB) followed by the EBUS and puncturing at least two different lymph node stations. RESULTS: Positive cytopathological verification of sarcoidosis in our study was obtained in 123 patients (92.5%). EBUS-TBNA was diagnostic in 116 patients (87.2%). EBB was positive in 26 patients (19.55%). Combination of EBUS-TBNA and EBB statistically increased diagnostic yield of sarcoidosis to 92.5%. Sensitivity of EBUS-TBNA with EBB was 93.9%, specificity 100%, PPV 100% and NPV 20%. CONCLUSIONS: Combining EBUS-TBNA from at least two lymph node stations and EBB increased diagnostic yield of sarcoidosis. Such diagnostic strategy had almost 93% of diagnostic yield in stage I and stage II of sarcoidosis. Taking into account the safety of the whole procedure with endobronchial ultrasonography combined with conventional endoscopy with EBB and its cost effectiveness, TBLB can be intended to diagnose stage III or IV of pulmonary sarcoidosis.

The evaluation of metallothionein expression in nasal polyps with respect to immune cell presence and activity.

BACKGROUND: The expression of metallothionein (MT) is involved in acquiring resistance to immune-mediated apoptosis; it is also a negative regulator of the immune response. Nasal polyps are typified by a resistance to immune-mediated apoptosis as well as by excessive immune cell infiltration. RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) is a membrane protein capable of inducing the apoptosis of CTLs and NK cells. The aim of the present study has been to explore the expression of metallothionein with respect to immune cell presence and immune cell activity. In our study, we identified immune cells using CD4 and CD68 antigen expression and evaluated their activity using CD25 antigen expression. We then analyzed metallothionein, RCAS1, CD25, CD4, and CD68 in a sampling of 50 nasal polyps using the immunohistochemistry method. We were able to divide the nasal polyps into three main groups according to their predominant immune cell infiltration: eosinophilic nasal polyps (21 cases), lymphocytic nasal polyps (17 cases), and neutrophilic nasal polyps (12 cases). RESULTS: In the present study, statistically significant differences between the MT expression in the epithelium and that in the stroma of the nasal polyps along with the accompanying alterations in activation markers on immune cells were found and the number of macrophages in both the eosinophilic and the lymphocytic nasal polyps was assessed. RCAS1-expressing macrophages were found only in the eosinophilic nasal polyps. CONCLUSION: MT expression seems to favor the survival of nasal polyp epithelial cells in the adjacent area of increasingly cytotoxic immune activity. RCAS1-expressing macrophages seem to participate in creating the immune suppressive microenvironment and so help to sustain local inflammation.

BRAF mutations in sporadic colorectal carcinoma from polish patients.

BRAF mutations are second to KRAS mutations in activation of the MAPK pathway in colorectal carcinoma cells. In addition to mutated KRAS, BRAF V600E mutation is associated with resistance to EGFR-targeted therapy in colorectal cancer; thus mutated BRAF might serve as a predictive factor. In this study, 163 routinely resected adenocarcinomas were screened for mutations in exons 11 and 15 of the BRAF gene. Only 6 (3.7%) tumours had a missense point mutation (G469A, D594G, G596R, K601N and twice V600E). The tumours were locally advanced with multiple metastases to lymph nodes. Mutations were associated with microsatellite instability (2 cases MSI-H, 2 cases MSI-L) and mutually exclusive with a mutated KRAS gene. In this sample set, mutations in the BRAF gene were more diverse and less frequent than usually reported.

Adenomatoid tumour of the adrenal gland: a case report and literature review.

Adenomatoid tumour (AT) is a rare, benign neoplasm of mesothelial origin, which usually occurs in the genital tract of both sexes. Occasionally these tumours are found in extra genital locations such as heart, pancreas, skin, pleura, omentum, lymph nodes, retroperitoneum, intestinal mesentery and adrenal gland. Histologically ATs show a mixture of solid and cystic patterns usually with focal presence of signet-ring like cells and scattered lymphoid infiltration. The most important thing about these tumours is not to mis-diagnose them as primary malignant or metastatic neoplasms. We present a case of an adrenal AT in a 29-year-old asymptomatic male. The tumour was an incidental finding during abdominal CT-scan for an unrelated condition. We also present a review of the literature concerning adrenal gland AT and give possible differential diagnosis.

The association between RCAS1 expression in laryngeal and pharyngeal cancer and its healthy stroma with cancer relapse.

BACKGROUND: The purpose of this study has been to establish the level of RCAS1 - a membrane protein expressed in various cancer cells and able to induce apoptosis of CTLs and NK cells in pharyngeal and laryngeal cancer and its clear surgical margin - with respect to clinicopathological features and to patient's follow up and evaluate its possible role in cancer relapse. METHODS: A total of 122 tissue samples were obtained: 51 samples from laryngeal and pharyngeal squamous cell carcinoma, 51 samples from the clear surgical margins of these tumors, and 20 tissue samples derived from the healthy mucous membranes of the upper respiratory tract mucosa of patients without cancerous tumors. Patients were observed for a total of 4 years following surgical treatment. The level of RCAS1 expression was assessed by immunohistochemistry and Western blot. RESULTS: RCAS1 was identified in all laryngeal and pharyngeal carcinomas and in almost all the clear surgical margin samples. The level of RCAS1 expression was significantly higher in the cancerous samples than in the clear surgical margins and was determined to be related to the grade of the cancer and the presence of lymph node metastases. In cases of cancer relapse, significantly higher levels of RCAS1 expression were observed in the clear surgical margins. CONCLUSION: Selective cytotoxic immune cell suppression concomitant with tumor growth and associated with RCAS1 expression seems to be an important event connected with cancer relapse.

Angiogenesis in the New Zealand obese mouse model fed with high fat diet.

BACKGROUND: Obesity and its complications lead to vascular injury, atherosclerosis, diabetes and pathological angiogenesis. One of the models to study the obesity and its entanglements is the New Zealand Obese mice model. Aim of this study was to check the effect of high fat diet on changes in biochemical parameters as well as on process of angiogenesis in NZO mice. METHODS: NZO mice were fed with standard (ST) or high fat (HF) diet for seven weeks. Body weight and serum biochemical parameters were monitored. The PECAM1 positive vessel-like structures immunostaining, as well as the gene expression of the matrigel penetrating cells by microarray (confirmed by real-time PCR method) were analyzed. RESULTS: Mice fed with HF diet developed obesity. Number of newly created vessels with lumen was correlated with hyperglycemia and animal weight gain. The number of PECAM1 positive cells in matrigel tended to increase during HF diet. Microarray results revealed changes in gene expression (activation of the oxidative stress and insulin resistance, inhibition of apoptosis and cell differentiation), however without markers of endothelial cell network maturation. CONCLUSION: Observed changes in the NZO mice on HF diet argue for the hyperglycemia related activation of angiogenesis, leading to the formation of pathological, immature network.

Primary cutaneous CD30+ lymphoproliferative disorder--a 10-year follow-up. A case report and differential diagnosis.

Primary cutaneous CD30+ lymphoproliferative disorders (LPDs) are the second most common group of primary cutaneous T-cell lymphomas (CTCLs). The spectrum of LPDs includes lymphomatoid papulosis (LyP), primary cutaneous anaplastic large cell lymphoma (C-ALCL) and borderline cases. The term "borderline lesions" refers to cases where histological features are similar to LyP, but clinically behave as C-ALCL, or to cases where histological features are typical for C-ALCL, but clinically behave as LyP. We present a clinical and morphological picture of LPD in a 57-year old patient treated in the Department of Oncology and of a relapse after ten years of follow-up and discuss clinical and morphological differential diagnosis and the significance of such diagnosis.