Anthocyanins Protect Hepatocytes against CCl4-Induced Acute Liver Injury in Rats by Inhibiting Pro-inflammatory mediators, Polyamine Catabolism, Lipocalin-2, and Excessive Proliferation of Kupffer Cells.

: This study examined the hepatoprotective and anti-inflammatory effects of anthocyanins from Vaccinim myrtillus (bilberry) fruit extract on the acute liver failure caused by carbon tetrachloride-CCl4 (3 mL/kg, i.p.). The preventive treatment of the bilberry extract (200 mg anthocyanins/kg, orally, 7 days) prior to the exposure to the CCl4 resulted in an evident decrease in markers of liver damage (glutamate dehydrogenase, sorbitol dehydrogenase, malate dehydrogenase), and reduced pro-oxidative (conjugated dienes, lipid hydroperoxide, thiobarbituric acid reactive substances, advanced oxidation protein products, NADPH oxidase, hydrogen peroxide, oxidized glutathione), and pro-inflammatory markers (tumor necrosis factor-alpha, interleukin-6, nitrite, myeloperoxidase, inducible nitric oxide synthase, cyclooxygenase-2, CD68, lipocalin-2), and also caused a significant decrease in the dissipation of the liver antioxidative defence capacities (reduced glutathione, glutathione S-transferase, and quinone reductase) in comparison to the results detected in the animals treated with CCl4 exclusively. The administration of the anthocyanins prevented the arginine metabolism's diversion towards the citrulline, decreased the catabolism of polyamines (the activity of putrescine oxidase and spermine oxidase), and significantly reduced the excessive activation and hyperplasia of the Kupffer cells. There was also an absence of necrosis, in regard to the toxic effect of CCl4 alone. The hepatoprotective mechanisms of bilberry extract are based on the inhibition of pro-oxidative mediators, strong anti-inflammatory properties, inducing of hepatic phase II antioxidant enzymes (glutathione S-transferase, quinone reductase) and reduced glutathione, hypoplasia of Kupffer cells, and a decrease in the catabolism of polyamines.

Pancreatic Polypeptide-Secreting Tumour of the Proximal Pancreas (PPoma)-Ultra Rare Pancreatic Tumour: Clinically Malign, Histologically Benign.

Background and objectives: Here we report a rare case of a pancreatic polypeptide-secreting tumour (PPoma) discovered by accident during an autopsy. These PPomas occur in less than 2% of all pancreatic neoplasms and are almost exclusively silent, i.e., they are non-functional. Symptoms arising from PPoma are due to its compression of surrounding tissue. Materials and methods: The autopsy was performed on a 68-year-old male diagnosed with multiple endocrine neoplasm type 1 (MEN1) due to the patient's sudden death. Results: A solitary, densely fibrotic, pink-brown tumour, 18 mm in size tumorous mass, was localised in the head of the pancreas. Microscopically, the tumour had a glandular structure with a tubuloacinar arrangement of the cells. Immunohistochemically, we detected strong PP (pancreatic polypeptide) intracytoplasmic activity and negative glucagon activity. The PPoma was located in the head of the pancreas, likely resulting in the obstruction of the main pancreatic and common bile duct. Conclusions: To the best of our knowledge, this is the first report suggesting the association of PPomas with MEN1. Also, the PPoma could be the cause of acute hemorrhagic pancreatitis due to its location.

Protective effects of anthocyanins from bilberry extract in rats exposed to nephrotoxic effects of carbon tetrachloride.

This study examined the nephroprotective effects of 15 different anthocyanins from the bilberry extract on the acute kidney injury caused by CCl4. The acute nephrotoxicity in rats was induced 24 h after the treatment with a single dose of CCl4 (3 mL/kg, i.p.).The nephroprotective effects of the anthocyanins were examined in the animals that had been given the bilberry extract in a single dose of 200 mg of anthocyanins/kg daily, 7 days orally, while on the seventh day, 3 h after the last dose of anthocyanins, the animals received a single dose of CCl4 (3 mL/kg, i.p.) and were sacrificed 24 h later. When the nephrotoxicant alone was administered, it resulted in a substantial increase of the pro-oxidative (TBARS, CD, H2O2, XO, and GSSG) and pro-inflammatory markers (TNF-α, NO, and MPO), as well as a noticeable reduction of the antioxidant enzymes (CAT, SOD, POD, GPx, GST, GR) and GSH when compared to the results of the control group. Moreover, the application of CCl4 significantly influenced a reduction of the renal function, as well as an increase in the sensitive and specific injury indicators of the kidney epithelial cells (ß2-microglobulin, NGAL, KIM1/TIM1) in the serum and urine of rats. The pretreatment of the animals poisoned with CCl4 with the anthocyanins from the bilberry extract led to a noticeable reduction in the pro-oxidative and pro-inflammatory markers with reduced consumption of the antioxidant defence kidney capacity, compared to the animals exposed to CCl4 alone. Anthocyanins have been protective for the kidney parenchyma, with an apparent absence of the tubular and periglomerular necrosis, severe degenerative changes, inflammatory mononuclear infiltrates and dilatation of proximal and distal tubules, in contrast to the CCl4-intoxicated animals. The nephroprotective effects of anthocyanins can be explained by strong antioxidant and anti-inflammatory effects achieved through the stabilization and neutralization of highly reactive and unstable toxic CCl4 metabolites.

Treatment of low-grade gastric MALT lymphoma using Helicobacter pylori eradication.

BACKGROUND/AIM: Lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) of the stomach usually occurs as a consequence of Helicobacter pylori (H. pylori) infection. The aim of this study was to investigate the long-term effect of treatment of low-grade gastric MALT lymphoma with the H. pylori eradication method. METHODS: In the period 2002-2012 in 20 patients with dyspepsia, mean age 55.1 years, the endoscopic and histologic diagnosis of gastric MALT lymphoma in the early stages were made. Histological preparations of endoscopic biopsy specimens were stained with hematoxyllineosin (HE), histochemical and immunohistochemical methods. RESULTS: Endoscopic findings of gastritis were documented in 25% of the patients, and 75% of the patients had hypertrophic folds, severe mucosal hyperemia, fragility, nodularity, exulcerations and rigidity. Histopathologically, pathognomonic diagnostic criterion were infiltration and destruction of glandular epithelium with neoplastic lymphoid cells, the so-called lymphoepithelial lesions. In all 20 patients H. pylori was verified by rapid urease test and Giemsa stain. After the triple eradication therapy complete remission of MALT lymphoma was achieved in 85% of the patients, with no recurrence of lymphoma and H. pyloi infection in the average follow-up period of 48 months. In 3 (15%) of the patients, there was no remission of MALT lymphoma 12 months after the eradication therapy. Of these 3 patients 2 had progression of MALT lymphoma to diffuse large-cell lymphoma. CONCLUSION: Durable complete remission of low-grade gastric MALT lymphoma is achieved in a high percentage after eradication of H. pylori infection, thus preventing the formation of diffuse large-cell lymphoma and gastric adenocarcinoma.

A retrospective review of 139 major and minor salivary gland tumors.

AIM: To describe demographic and histomorphological characteristics of 139 patients with epithelial salivary gland tumors in the Southeastern Serbia population. METHODS: A total number of 139 patients with epithelial tumors arising in major and minor salivary glands in the period 2010-2012 was evaluated. After standard tissue proceeding, the routine haematoxylin-eosin (HE) and histochemical alcian blue-periodic acid-Schiff (AB - PAS) methods were used for histomorphological examination. RESULTS: Among 139 patients, 102 (73.38%) had benign, and 37 (26.62%) malignant tumors. The majority of tumors were localized in the parotid gland, in 117 (84.17%) patients. Among benign tumors there were 50 (49.02%) pleomorphic adenoma, 48 (47.06%) Warthin's tumor, two (1.96%) myoepithelioma, and two (1.96%) oncocytoma. In the group of malignant tumors the most common was mucoepidermoid carcinoma, in 12 (32.43%) patients, carcinoma ex pleomorphic adenoma in six (16.22%), adenoid cystic carcinoma in five (13.51%), and oncocytic carcinoma in three (8.11%) patients. CONCLUSION: Benign tumors were more common than malignant ones, with predominance of pleomorphic adenoma. Malignant tumors are less common than benign in the large salivary glands, and more common in the minor salivary glands. Histochemical AB-PAS method helps in the diagnosis of mucinous salivary gland carcinoma.

Comparative analysis of clinicopathological and immunohistochemical characteristics of Merkel cell carcinoma.

PURPOSE: To investigate both the clinicopathological and immunohistochemical characteristics of a very rare skin cancer - Merkel cell carcinoma (MCC) - and to review the relevant literature. METHODS: The study group was composed of 12 patients, with mean age 53.08±10.26 years. Multiple subcutaneous masses and lymph node metastases were surgically removed. Paraffin blocks of formaldehyde-fixed tumor tissue were cut and stained for histological, histochemical and immunohistochemical studies. The following antibodies (Dacopatt) were used: Chromogranin A, CK20, CK7, Melan A, CD20 and CD45Ro. RESULTS: The tumors involved the dermis while sparing the epidermis. The most frequently affected sites were sun-exposed skin (8 patients on the head and neck) and the most common histological subtype of MCC was the intermediate variant. Six patients had lymph node metastasis and 2 had locoregional recurrences. Haematogenous lung metastases of MCC and primary located in the trunk were found only in our youngest patient (36-year-old). Immunostaining revealed positive reactivity for neuroendocrine and epithelial markers and negative reactivity for melanoma, B/T lymphomas and small cell metastatic lung carcinoma. CONCLUSION: MCC is a rare malignant primary cutaneous neoplasm with epithelial and neuroendocrine differentiation, demanding wide local excision. The pathological differential diagnosis includes basal cell carcinoma, melanoma, lymphoma, and metastatic small cell lung carcinoma. The diagnosis of MCC is possible only immunohistochemically, by using the wide spectrum of antibodies, characteristic of microscopically similar tumors.

Peutz-Jeghers syndrome: quantitative study on enterochromaffin cells in hamartomatous intestine polyps.

INTRODUCTION: Peutz-Jeghers (PJ) syndrome is a rare familial disorder with the autosomal transmission characterized by multiple intestinal polyps, mucocutaneous pigmentation and increased incidence of various malignancies. Some clinical manifestations of PJ syndrome may be associated with the serotonin secretion from the enterochromaffin cells (EC). OBJECTIVE: Since no data have been reported so far regarding EC cells in PJ polyps, the aim of our study was to quantitatively investigate EC population in hamartomatous intestinal polyps in patients with the PJ syndrome. METHODS: The samples of surgically removed PJ polyps from family members with the PJ syndrome were collected during 34-year follow-up period. Formalin-fixed paraffin-embedded specimens of twenty-one PJ polyps were stained with HE, AB-PAS, Van Gieson, Fontana-Masson, FIF and Grimelius. For immunohistochemical analysis, the following antibodies were used: chromogranin A, serotonin, Ki-67, desmin, vimentin and cytokeratin in order to eliminate differential diagnostic possibilities and to confirm diagnosis of PJ polyps. RESULTS: Strong EC cell hyperplasia was observed within the tissue of the investigated polyps. Statistical analysis demonstrated significantly higher content of EC cells in PJ polyps than in the normal ileal mucosa. CONCLUSION: Marked hyperplasia of EC cells within the PJ polyps may be the most important contributor to functional disorders in patients with the PJ syndrome.

Focal neuroendocrine differentiation in prostatic gland carcinoma with basaloid pattern.

INTRODUCTION: Prostatic gland basal cell proliferations exhibit morphological continuum ranging from basal cell hyperplasia to basal cell carcinoma. In the following report, we described clinical features, morphological spectrum, neuroendocrine differentiation and histogenesis of prostatic gland basal cell carcinoma in our patient. CASE REPORT: Hematoxylin-eosin (HE), Alcian blu-periodic acid schiff (AB-PAS) at pH 2.5 stained sections and the avidin-biotin-peroxidase complex (ABC), were performed on prostate gland paraffin-embedded tissue. Monoclonal antibodies directed against cytokeratin (34betaE12) which selectively stains basal cells, prostate specific antigen (PSA), chromogranine A, neuron-specific enolase (NSE), synaptophysin and CD56, were used. Basal cell proliferations exhibited a morphological continuum ranging from basal cell hyperplasia to prostatic gland carcinoma. In these prostatic lesions, positive reactivity was demonstrated for 34betaE12 and CD56. These findings indicate that the basaloid cells of basal cell hyperplasia, florid basal cell hyperplasia, atypical basal cell hyperplasia and basal cell carcinoma are derived from basal cells of the normal prostate gland suggesting a continuum in the progression of hyperplasia to benign and then malignant neoplasia. The presence of CD56 protein in the discovered lesions may be related to their neuroendocrine differentiation. CONCLUSION: The fact, that our patient was well six years after the radical prostatectomy supports the belief of some authors that basal cell carcinoma represents a low grade carcinoma with an excellent prognosis.

Celiac disease prevalence in children and adolescents with type 1 diabetes from Serbia.

BACKGROUND: The association between celiac disease (CD) and type 1 diabetes mellitus (T1DM) is well known. Up to now, CD prevalence in children and adolescents with T1DM in Serbia has not been reported. The aim of the present study was to determine CD prevalence and its clinical manifestations in patients with T1DM. METHODS: One hundred and twenty-one patients (70 girls, 51 boys; mean age, 10.8 years) with T1DM (mean duration of diabetes, 3.4 years) and 125 control group participants (75 girls, 50 boys; mean age, 10.4 years) were tested for CD on tissue transglutaminase antibodies (tTG). In seven serologically positive T1DM patients endoscopic small bowel biopsies were taken and examined on histopathology. In all patients with CD and T1DM age, duration of T1DM, height for age, body mass index, glycosylated hemoglobin and clinical symptoms were noted. RESULTS: Nine patients with T1DM were positive on IgA tTG antibodies. In seven of them small bowel biopsy was performed, and all were proven to have CD on histopathology. The prevalence of biopsy-proven CD in children and adolescents with T1DM was significantly higher in the study group compared to controls (5.79%. vs 0.8%, P < 0.05). CONCLUSION: The significantly higher prevalence of CD in children with type 1 diabetes, in accordance with the large volume of data published in the literature, underlines the need for yearly screening of CD in patients with diabetes in order to promptly start a gluten-free diet when appropriate.

[Preoperative identification of bleeding site caused by angiodysplasia of the small bowel by means of selective arteriography and application of methylene blue].

BACKGROUND: Small bowel hemorrhages are rare and account for 2-10% of all gastrointestinal bleedings. In case that surgery is necessary, identification of the bleeding site is the most important problem. CASE REPORT: We presented here the case of a 65-year old man, admitted for urgent care of massive lower gastrointestinal bleeding. After reanimation and normalization of vital parameters, selective arteriography was done. A contrast extravasation site was identified at the level of jejunal branches of a. mesenterica superior and labeled by means of methylene blue application. Immediately after we performed conservative resection of the labeled jejunal loop in 10 cm length and terminoterminal anastomosis. The preparation was sent for histopathologic examination--small bowel angiodysplasia was identified. The patient was monitored in three month intervals in the next two years and new bleeding events were not observed. CONCLUSION: Bleeding caused by small bowel angiodysplasia is a significant diagnostic problem in cases in whom urgent surgery is required. Combined preoperative selective arteriography and methylene blue application make possible precise identification of the bleeding site as well as conservative small bowel surgery, avoiding thus the risk and danger of malabsorption syndrome.

Malignant schwannoma of the pancreas involving transversal colon treated with en-bloc resection.

Pancreatic schwannoma is a very uncommon tumor of the pancreas, with only 27 cases reported. Most pancreatic schwannomas are benign, with only four malignant tumors reported. We describe a case of giant malignant schwannoma of the pancreatic body and tail, which involved the transverse colon. The tumor was treated successfully with en bloc distal splenopancreatectomy and colon resection. This is believed to be the first reported radical operation for malignant schwannoma of the pancreatic body, with infiltration of the transverse colon, with excellent long-term results. The patient is alive and well 28 mo after the operation. The authors conclude that pancreatic schwannomas should be considered in the differential diagnosis of cystic neoplasms of the pancreas, although the diagnosis can only be confirmed by microscopic examination. In the case of the benign tumors, local excision is adequate, but in the case of malignant schwannoma, oncological standards must be fulfilled.

[Endoscopic resection of inflammatory gastric fibroid polyp--a case report].

INTRODUCTION: An inflammatory fibroid polyp is a rare condition with frequent localisation in the antrum of the stomach. Because of the localisation in the bottom parts of the mucosa and submucosa, a histological diagnosis is difficult to establish at endoscopic biopsies. So, a correct diagnosis is histologically possible after surgical excision which is a common manner of treatment. Many authors have shown that endoscopic removal of an inflammatory fibroid polyp is possible. CASE OUTLINE: We are presenting a case of complete endoscopic resection of an inflammatory fibroid polyp of the stomach antrum in a 72-year-old patient. He comllained of nausea, vomiting and loss of appetite. The polyp endoscopically looked like flat elevation with central umbilication, 16 mm in diameter and it was localised praepylorically. Pathohistologically, areas of severe epithelial dysplasia were verified at endoscopic biopsies which suggested early gastric cancer. Helicobacter pylori (H. pylori) infection was verified histologically and with rapid urease test. The patient was treated with triple eradication H. pylori therapy. Two months after the therapy, we decided to perform endoscopic resection of the polyp. We applied a suction technique of mucosal resection by which the polyp was completely resected. After complete endoscopic removal, the final diagnosis of an inflammatory fibroid polyp localised in the submucosa and mucosa of the antrum of the stomach was histologically made. On follow-ups, after 3, 6 and 12 months, there were no endoscopic and histological signs of either inflammatory fibroid polyp relapse, or recurrence of H. pylori infection. The patient had no dyspeptic symptoms. CONCLUSION: We recommend an endoscopic method of resection as a therapy of choice for an inflammatory fibroid polyp of the stomach.

[Role of mucinohistochemical analysis in verification of malignant potential of colorectal adenoma].

BACKGROUND/AIM: It is a general opinion that the biggest number of colorectal carcinoma (CRC) develops with the malignant alteration of colorectal adenomas (adenoma carcinoma sequence). Malignant potential of adenoma is in the function of its size, histological structure and the grade of dysplasia. An adequate analysis of colorectal adenomas helps their definition and timely removal, which significantly decreases carcinoma incidence. Mucin analysis can provide results which support the adenoma carcinoma sequence theory. The aim of the study was to present histochemical characteristics of adenomas and emphasize the significance of these analyses for the precise adenoma definition. METHODS: This prospective study included analysis of 117 colorectal adenomas obtained by transcolonoscopic biopsy or polipectomy in 82 patients. The biopsy samples and the removed polyps were coloured by hematoxilin-eozin (HE) staining and histochemical stainings by Periodic Acid Schiff (AB-PAS) and High Iron Diamine/Alcian blue (HID-AB; pH = 2.5) to prove mucins. RESULTS: Sulphomucins are more often found in adenomas of the left colon than of the right one (p < 0.001). Sialomucins are more reactive in adenomas of the right colon (p < 0.001). Sulphomucins are more reactive in adenomas of < 10 mm diameter (p < 0.05). The reactivity of all mucins is in negative correlation with the degree of dysplasia (p < 0.001). There is a significant difference in the reactivity of sialomucins in adenomas of the same histological type but different degree of dysplasia (chi2 = 25,743, df = 6, p < 0.001). CONCLUSION: The difference in the presence of mucins between adenomas of the left and right colons supports the theory of adenoma carcinoma sequence, since adenomas of the right colon are more protected by sialomucins and therefore less malignantly altered, which is consistent with CRC topography. Histochemical analysis of colorectum is significant for more accurate gradation of dysplasia and confirmation of malignant alteration. The negative correlation between the degree of dysplasia and the production of mucin indicates the significance of the degree of dysplasia in malignant potential of colorectal adenomas.

An immunohistochemical study of Ki-67 in colorectal adenoma.

INTRODUCTION: The expression of the human Ki-67 protein is strictly associated with cell proliferation. The cell proliferation and kinetics of normal gastrointestinal tract are well known but the cell kinetics of adenoma in gastrointestinal tract is poorly understood. The aim of this study was to provide an immunohistochemical evaluation of the expression of Ki-67 antigen in different colorectal adenomas. MATERIAL AND METHODS: In the prospective study, we analyzed 65 colorectal adenomas. RESULTS: Proliferation indexes were found to steadily increase from normal mucosa to adenoma (p < 0.05). There was also a difference in the expression of Ki-67 between adenomas smaller and larger than 10 mm (p < 0.05). The expression of Ki-67 was strongly more often in villous adenomas as opposed to the tubular adenomas (p < 0.05). Reaction for Ki-67 were strong in adenomas with high grade atypia compared to low grade adenomas (p < 0.001). CONCLUSION: The expression of Ki-67 clearly shows differences between advanced and nonadvanced adenomas. The strong reactions of Ki-67 in adenomas with severe dysplasia show a close association with colorectal carcinoma. The immunohistochemical analysis (Ki-67) may be included as a part of routine pathological evaluation with conventional prognostic factors.

[Primary colorectal lymphomas].

BACKGROUND/AIM: Colorectal lymphoma is a rare tumor representing 1.4% of human lymphomas, 10-20% of gastrointestinal lymphomas, namely 0.2-0.6% of all malignancies in the colon. The aim of this study was to review clinical characteristics of primary colorectal lymphoma and overall survival. METHODS: A detailed analysis of 16 surgically treated patients included patients age, symptoms and signs, tumor site, type of surgery, histopathologic findings, diagnosis of the disease, disease stage, type of surgery related to the degreee of emergency (elective or urgent), applied adjuvant therapy, patient follow-up and treatment outcomes. Survival was expressed by the Kaplan-Meier curve, while the difference in survival among the two groups by the Log-rank test. RESULTS: The all patients were on an average followed-up for a median of 29 months (range 2-60 months), while those with chemotherapy 48 months (range 4-60 months). An overall mean survival time was 38.65 months. CONCLUSION: Primary colorectal lymphoma is a rare malignant tumor of the large bowel. Therapy usually involves resection of the affected colon or rectum and regional lymphovascular structures, followed by adjuvant therapy. Survival period is short and, therefore, timely diagnosis is crucial in early disease stages when the probability of cure is high.

An approach to malignant mammary phyllodes tumors detection.

BACKGROUND/AIM: Mammary phyllodes tumors (MPT) are uncommon fibroepithelial (biphasic) neoplasms whose clinical behavior is difficult to predict on the basis of histological criteria only. They are divided into benign, borderline malignant and malignant groups. Sometimes it appears difficult to distinguish these tumors from other types of soft tissue sarcomas. Because of the relatively scant data on the role of biological markers in MPT histogenesis, we have decided to undertake the following study, trying to shed more light on the issue by investigating the following elements that make up MPT: their histological patterns, biological behavior, enzymohistochemical, histochemical and immunohistochemical characteristics (ICH) together with the mast cell analysis. METHODS: We examined the biopsy material of 35 MPT in our laboratory. Enzymohistochemistry was performed on frozen sections (method of Crowford, Nachlas and Seligman). The used methods were classical hematoxylin-eosin (H & E); histochemical Massontrichrome, Alcian-blue, Periodic acid Schiff and immunohistochemical LSAB2 method (DacoCytomation). Ki-67, c-kit, vimentin, estrogen receptor (ER), progesterone receptor (PR) and Her-2 oncoprotein immunohistochemistry was performed on all tumors. RESULTS: The patients were ranged per age from 30--62 years (mean 43.3 years, median 39 years). A total of 35 cases of MPT were included: 20 benign (57%), 6 borderline malignant (17%) and 9 malignant (26%). Twenty-two patients (62.8%) underwent segmental mastectomy, while 13 (37.2%) had total mastectomies. Twenty-eight patients had negative surgical margins at original resection. The mean size of malignant MPT (7.8 cm) was larger than that of benign MPT (4.5 cm). Significant features of the malignant MPT were: stromal cellularity, stromal cellular atypism, high mitotic activity, atypic mitoses, stromal overgrowth, infiltrative tumor contour and heterologous stromal elements. Benign MPT showed strong enzymohistochemical Leucine Amino Peptidase (LAP) activity in both epithelial and stromal components while it was weak or absent in the epithelial parts of the malignant tumors. Acid mucopolysacharides were present in the stromal component of all types of these tumors. Benign MPT had a lower Ki-67 than did borderline malignant MPT (4 versus 28). Malignant MPT had a greater than 8-fold higher Ki-67 activity than did benign tumors (35 versus 4). Intracyto-plasmatic c-kit expression was associated with a pathological diagnosis of malignant MPT, correlating with increasing grade (p < 0.05). In hypercellular stroma of borderline malignant and especially malignant forms of MPT, high activity of ER in mast cells was confirmed. Oncoprotein Her-2 activity, mostly in epithelial components, correlated with the degree of malignant progression of MPT (p < 0.05). CONCLUSION: Besides the well-known malignant features additional parameters have been found to be high Ki-67 and c-kit stromal expressions, and weak LAP activity in the epithelial part of malignant MPT, as well as mast cells with a high expression of ER.

Clinical, histopathological and immunohistological study of lymphoid disorders in the parotid gland of patients with Sjögren's syndrome.

BACKGROUND/AIM: Sjögren's syndrome is a chronic autoimmune systemic disease characterized by polyglandular tissue destruction, leading to keratoconjunctivitis sicca and xerostomia. These patients have 44-fold increased risk of developing salivary gland lymphoma, of which 80% are marginal zone (MALT) type. Having in mind that criteria for distinguishing benign lymphoepithelial lesions from MALT lymphoma are obscure, the aim of this study was to provide practical information that could be integrated into diagnostic practice. METHODS: Among 32 parotidectomies, 27 cases were identified as having benign lymphoepithelial disorders and 5 cases low grade MALT lymphoma. Histological sections were stained routinely with hematoxylin and eosin (H&E and special stains. Immunohistochemical study was performed by LSAB2 method, by using primary antibodies for CD20, CD3, Kappa and Lambda light chains and Cytokeratin (Dako Denmark). RESULTS: The 27 patients with Sjögren's sialoadenitis (22 women and 5 men), and 5 patients with MALT lymphoma (only women) were included in this analysis. According to the Ann Harbor Classification, all patients with MALT lymphoma had stage IE. Both groups of patients had an indolent clinical course, except permanent, rapid parotid enlargement in the patients with MALT lymphoma. Histologically, the periductal lymphoid infiltrate, gradually extended to the acini, completely replacing them by a sea of polyclonal lymphocytes, immunoblasts, germinal centers and plasma cells (confirmed immunohistochemically), but sparing the ducts and preserving lobular appearance. The histological feature of salivary gland MALT lymphoma included heterogeneous B-cell infiltrate that totally or subtotally had effaced the normal glandular structure. Malign lymphoepithelial lesions, representing infiltration of the ductal and epithelial structures by monoclonal neoplastic B-cells, positive for CD20, were highlighted by antibody to cytokeratin. CONCLUSION: The optimal diagnosis of salivary gland MALT lymphoma requires careful integration of clinical, morphological and immunohistochemical results.

[Lynch syndrome I: a case report].

INTRODUCTION: Hereditary nonpolyposis colorectal cancer (HNPCC), also known as Lynch syndromes I and II, accounts for about 5-8% of colorectal cancers. Lynch syndrome I is an autosomal dominant inherited disorder characterized by early onset of colorectal cancer, predominance of proximal and multiple tumors, and microsatellite instability. In order to identify HNPCC, the international "Amsterdam criteria" have been used. CASE REPORT: The proband was a 40-year-old male who was admitted to hospital with a diagnosis of advanced rectal cancer. Left colectomy was carried out. A histopathologic diagnosis of poorly differentiated adenocarcinoma of clinical stage Dukes C was made. The family talking was done and it was revealed that the proband had five family members (one of first degree relative) with colorectal cancer, and two successive generations affected. All malignancy were diagnosed before 45 years of age. In one family member, metachronous transverse cancer was revealed 12 years after surgery for cecal adenocarcinoma. DISCUSSION AND CONCLUSION: The main molecular cause for HNPCC is constitutional mutation in one of the mismatch repair (MMR) genes that regulate the excision of errors occurring during DNA replication. The most often are mutations of MLH1 and MSH2 genes, and microsatellite instability is present in about 90-95% HNPCC. In this report, we present a case of an HNPCC patient who met the Amsterdam criteria for Lynch syndrome I. Family members that fulfil the Amsterdam criteria should be investigated for mutation in MMR genes. The genetic tests are not routinely available, so colonoscopic screening of all asymptomatic family members older than 25 has been recommended.

Computer-assisted quantitative analysis of Ki-67 antigen in dysplasia--associated lesions or masses in ulcerative colitis.

BACKGROUND/AIM: The aim of this study was to apply computer-assisted methodology in assessment of Ki-67 positivity in "adenoma-like" dysplasia associated lesions or masses (DALMs), and carcinoma in ulcerative colitis (UC), and to determine a new approach to grading of Ki-67 staining intensity. METHODS: Immunohistochemical slides were quantitatively analyzed for estimation of proportion and intensity of Ki-67 positive-stained cells in a total of 50 "adenoma-like" DALMs (27 with low-grade dysplasia and 23 with high-grade dysplasia), and 17 adenocarcinomas associated with UC. The four grades of immunohistochemical staining intensity were established by an automated classification of nuclear optical densities. RESULTS: The Ki-67 labeling index (LI) in low-grade dysplasia was significantly lower than in high-grade dysplasia, and carcinoma (p < 0.001). The Ki-67 LI of carcinomas was not significantly different from the value obtained in high-grade dysplasia (p > 0.05), however having the difference in percentage values of the moderate stained nuclei (p < 0.05). The overall average values of chromogene nuclear optical density, showed statistically significant differences between DALMs and carcinoma (p < 0.05), although not between normal mucosa and low-grade dysplasia (p > 0.05). CONCLUSION: The obtained results imply, according to the overall percentage of labeled nuclei, that high-grade dysplasia is very close to carcinoma, while there is the difference in the percentage of moderately stained nuclei. We showed that Ki-67 positivity have a different internal distribution which could be useful in analysing these lesions. These findings also, indicate the important biological differences between low-grade dysplasia and carcinoma in UC, and a low proliferative potential of the former. Automated image analysis permits an objective [corrected] estimation of Ki-67 immunohistochemical staining in UC-associated dysplasia and carcinoma.