Hepatoprotektivni učinak etil-acetatnog ekstrakta biljke Teucrium polium L. na jetru štakora oštećenu tetraklormetanom

Hepatoprotective activity of the ethyl acetate extract of Teucrium polium (L.) was investigated using rats with CCl4-induced liver damage. Specific biochemical parameters (glutathione peroxidase, superoxide dismutase, reduced glutathione and total antioxidative status) were estimated in blood and in liver homogenate. Lipid peroxidation in CCl4-intoxicated rats was evidenced by a marked increment in the levels of thiobarbituric acid reactive substances. Histopatological examinations of the liver were undertaken to monitor the liver status. Silymarin was used as a standard to compare the hepatoprotective activity of the extract. Some biochemical parameters in groups treated with the Teucrium polium extract at a dose of 25 mg kg-1, showed significantly different values than that of the CCl4-treated group. The liver biopsy of all experimental rat groups treated with the Teucrium polium ethyl acetate extract showed significant restoration of the normal histomorphological pattern of liver cells. The study substantiates the potential hepatoprotective activity of the ethyl acetate extract of Teucrium polium L.Hepatoprotektivno djelovanje etil-acetatnog ekstrakta biljke Teucrium polium L. ispitivano je na štakorima čija je jetra oštećena tetraklormetanom. Specifični biokemijski parametri (glutation peroksidaza, superoksid dismutaza, reducirani glutation i ukupni antioksidativni status) procijenjeni su u krvi i homogenatu jetre. Lipidna peroksidacija u štakora intoksiciranih tetraklormetanom dokazana je povećanjem koncentracije reaktivnih derivata tiobarbiturne kiseline. Stanje jetre procijenjeno je histopatološkim ispitivanjima. U ispitivanjima hepatoprotektivnog djelovanja ekstrakta Teucrium polium upotrebljen je silimarin kao standard. Neki biokemijski parametri u skupini životinja tretiranih ispitivanim ekstraktom u dozi od 25 mg kg-1 imali su značajno različite vrijednosti od skupine tretirane s CCl4. Biopsija jetre u svim ispitivanim životinjama tretiranim ekstraktom pokazala je uspostavu normalnih histomorfoloških uvjeta u stanicama, što ukazuje na hepatoprotektivno djelovanje biljke Teucrium polium

Inflammatory Pseudotumor/Inflammatory Myofibroblastic Tumor of Spleen – A Case Report

BACKGROUND: Splenic inflammatory pseudotumor (IPT)/inflammatory myofibroblastic tumor (IMT) is a rare pseudotumor/tumor of unknown origin, which is usually benign, although atypical and aggressive cases have been reported. It is a lesion composed of proliferated myofibroblastic cells (hence IMT by some authors) with admixed pleomorphic inflammatory cells of varying proportions. CASE REPORT: Herein, we report a case of 61-year-old male patient with ill-defined abdominal discomfort and no other symptoms and signs. Clinical and imaging investigations revealed a mass in the spleen that was equivocally interpreted as secondary neoplasm, although primary neoplasm of the spleen was not excluded by the radiologists. Splenectomy was performed and on gross examination a well demarcated greyish-livid tumor measuring 3.5 cm × 3 cm × 3 cm was discovered. Microscopic examination showed proliferation of loosely arranged spindle cells admixed with inflammatory cells (histiocytes, lymphocytes, neutrophils, eosinophils, occasional plasma cells, and/or plasmacytoid cells) with varying density and multifocal clustering, multifocal hemorrhage, and fibrinoid-like deposition. We performed additional histochemical and immunohistochemical stainings which were consistent with the diagnosis of IPT/IMT. Next-generation sequencing (TruSight Tumor 15) showed common TP53 polymorphism (c.215C>G; p.Pro72Arg) along with several intronic and synonymous single nucleotide variations (SNVs), as well as five low confidence missense SNVs. Sixteen months after the operation the patient has uneventful follow-up. CONCLUSION: Although the incidence of IPT/IMT is low, awareness of its existence is necessary. The prognosis is favorable following splenectomy in most cases. Careful microscopic examination of the specimen is mandatory, due to possible misdiagnosis. We believe that extensive NGS analysis on archive samples would provide more data about the spectrum of possible genetic changes in lesions like IPM/IMT

Extranodal Diffuse Large B-Cell Lymphoma of the Small Bowel in Female Patient Causing Intestinal Obstruction: A Case Report

Diffuse large B cell lymphoma is the most common extranodal non-Hodgkin lymphoma of the small intestine accounting for more than 50% of cases. Forty percent of these cases initially present with small bowel obstruction. Therefore, the diagnosis is usually established after surgery for bowel obstruction. The treatment is then continued with a certain chemotherapy regimens. We present a case of a 46-years-old female patient with signs of small bowel obstruction due to previously undiagnosed diffuse large B-cell lymphoma. Postoperatively, the patient was treated with 7 cycles of R-CHOP protocol and complete response was achieved in the short follow-up period

Correlation of Immunohistochemistry and Fluorescence in Situ Hybridization for HER-2 Assessment in Breast Cancer Patients: Single Centre Experience

BACKGROUND: Accurate assessment of HER-2 is imperative in selecting patients for targeted therapy. Most commonly used test methods for HER-2 are immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH). We evaluated the concordance between FISH and IHC for HER-2 in breast cancer samples using Food and Drug Administration approved tests.MATERIAL AND METHODS: Archived paraffin tissue blocks from 73 breast cancer patients were used. HER-2 immunostaining was performed using Ventana anti–HER-2 monoclonal antibody. The FISH assay was performed using PathVysion™ HER-2 DNA Probe Kit.RESULTS: Of the 73 cases 68.5% were IHC 0/1+, 15.07% were IHC 2+ and 16.44% were IHC 3+. Successful hybridisation was achieved in 72 cases. HER-2 FISH amplification was determined in 16.67% cases. Ten IHC 3+ and two IHC 2+ cases were FISH positive. Two of the IHC 3+ cases were FISH negative. Concordance rate was 100%, 18.18% and 83.33% for IHC 0/1+, 2+ and 3+ group, respectively. Total concordance was 84.72%, kappa 0.598 (p < 0.0001). The sensitivity of IHC in detecting IHC 2+ and IHC 3+ cases was 16.7% and 83.3%, and the specificity was 85% and 96.67%, respectively.CONCLUSION: The consistency between the methods was highest for IHC negative and lowest for IHC equivocal cases. The immunohistochemistry showed high sensitivity for IHC 2+/3+ cases and high specificity for IHC 3+ cases. Our results support the view that false-positive rather than false-negative IHC results are a problem with HER-2/IHC testing, and that IHC should be used as an initial screening test, but IHC 2+/ 3+ results should be confirmed by FISH

Presentation of the Molecular Subtypes of Breast Cancer Detected By Immunohistochemistry in Surgically Treated Patients

INTRODUCTION: The detection of estrogen, progesterone and HER-2 neu receptors on the surface of the tumour cell is a significant prognostic factor, alone or in combination. The presence or absence of receptors on the surface of the tumour cell is associated with the conditional gene expression in the tumour cell itself. Based on these genetically determined expressions of the tumour cell, five molecular subtypes of breast cancer have been classified on the St. Gallen International Expert Consensus in 2011 that can be immunohistochemically detected, with each subtype manifesting certain prognosis and aggression.AIM: Analyzing the presentation of molecular subtypes of breast cancer that are immunohistochemically detected in surgically treated patients at the Clinic for Thoracic and Vascular Surgery.MATERIAL AND METHODS: We used the international classification on molecular subtypes of breast cancer which divides them into: Luminal A (ER+ and/or PR+, HER-2 negative, Ki-67 < 14%), Luminal B with HER-2 negative (ER+ and/or PR+, HER-2 negative, Ki-67 ≥ 14%), Luminal B with HER-2 positive (ER+ and/or PR+, HER-2+, any Ki-67), HER-2 enriched (ER-, PR-, HER-2+), and basal-like (triple negative) (ER-, PR-, HER-2 negative, CK5/6+ and/or EGFR+). A total of 290 patients, surgically treated for breast cancer, were analysed during 2014.RESULTS: In our analysis, we found that Luminal A was present in 77 (26.55%) patients, Luminal B HER-2 negative was present in 91 (31.38%) patients, Luminal B HER-2 positive was present in 70 (24.14%) patients, HER-2 enriched was present in 25 (8.62%) patients and basal-like (or triple negative) was present in 27 (9.31%) patients.CONCLUSION: Detecting the subtype of breast cancer is important for evaluating the prognosis of the disease, but also for determining and providing an adequate therapy. Therefore, determining the subtype of breast cancer is necessary for the routine histopathological assay

The Spectrum of Histopathological Changes in the Renal Allograft - a 12 Months Protocol Biopsy Study

INTRODUCTION: Renal transplantation became a routine and successful medical treatment for Chronic Kidney Disease in the last 30 years all over the world. Introduction of Luminex based Single Antigen Beads (SAB) and recent BANFF consensus of histopathological phenotypes of different forms of rejection enables more precise diagnosis and changes the therapeutic approach. The graft biopsies, protocol or cause, indicated, remain a golden diagnostic tool for clinical follow up of kidney transplant recipients (KTR).AIM: The study aimed to analyse the histopathological changes in renal grafts 12 months after the surgery in KTR with satisfactory kidney function.MATERIAL AND METHODS: A 12-month protocol biopsy study was performed in a cohort of 50 Kidney transplant recipients (42 from living and 8 from deceased donors). Usual work-up for suitable donors and recipients, standard surgical procedure, basic principles of peri and postoperative care and follow up were done in all KTR. Sequential quadruple immunosuppression including induction with Anti-thymocyte globulin (ATG) or Interleukin-2R antagonist (IL-2R), and triple drug maintenance therapy with Calcineurin Inhibitors (CNI), Mycophenolate Mofetil (MMF) and Steroids were prescribed to all pts. Different forms of Glomerulonephritis (16), Hypertension (10), End Stage Renal Disease (13), Hereditary Nephropathies (6), Diabetes (3) and Vesicoureteral Reflux (2) were the underlying diseases. All biopsies were performed under ultrasound guidance. The 16 gauge needles with automated “gun†were used to take 2 cores of tissue. The samples were stained with HE, PAS, Trichrome Masson and Silver and reviewed by the same pathologist. A revised and uploaded BANFF 2013 classification in 6 categories (Cat) was used.RESULTS: Out of 48 biopsies, 15 (31%) were considered as normal, 4 (8%), Borderline (BL-Cat 3), 5 (10%) as Interstitial Fibrosis/Tubular Atrophy (IF/TA-Cat 5), 5 (10%) were classified as non-immunological (Cat 6), 2 as a pure antibody-mediated rejection (ABMR-Cat 2) and T-cell Mediated Rejection (TCMR-Cat 4). The remaining 17 samples were classified as a “mixed†rejection: 7 (41%) ABMR + IF/TA, 5 (29%) ABMR + BL + IF/TA, 2 (11%) BL + IF/TA, 1 (5%) ABMR + BL, 1 (5%) ABMR + TCMR and 1 (5%) TCMR +  IF/TA. The mean serum creatinine at the time of the biopsy was 126.7 ± 23.4 µmol/L, while GFR-MDRD 63.4 ± 20.7 ml/min, which means that the majority of the findings were subclinical. Among the non-immunological histological findings (Cat 6), 3 cases belonged to CNI toxicity, 1 to BK nephropathy and 1 to recurrence of the primary disease.CONCLUSION: Our 12-month protocol biopsy study revealed the presence of different forms of mixed subclinical rejection. Use of recent BANFF classification and scoring system enables more precise diagnosis and subsequently different approach to the further treatment of the KTR. More correlative long-term studies including Anti HLA antibodies and Endothelial Cell Activation- Associated Transcripts (ENDAT) are needed

Tumour Lymphocytic Infiltration, Its Structure and Influence in Colorectal Cancer Progression

BACKGROUND: The role of the immune system in the control of tumour progression has been stressed, recently. Many studies indicate the fact that the immune system can prevent tumour progression in several types of human malignant neoplasms including colorectal cancer. According to some authors, a higher density of “tumour-associated lymphocytes†(TAL), in malignant neoplasms, correlate with prolonged survival of patients.AIM: This study aims to determine the structure and the influence of the immune cells, TAL, in the progression of colorectal cancer (CRC).PATIENTS AND METHODS: The study included 103 patients with CRC operated at the University Clinic of Digestive Surgery in Skopje, whose operative material was analysed at the Institute of Pathology, Medical Faculty in Skopje. The structure of tumor-associated cells and their density were determined and were correlated with neoplasm’s grade, local growth (T), positive lymph nodes, lymphatic invasion and stage of the disease.RESULTS: CD4+, CD8+ and CD20+ lymphocytes (Ly) were found in TAL. The density of TAL was significantly different in neoplasms with different T status, lymphatic invasion, patients with and without nodal metastasis and patients with a different stage of the disease. The density of CD4+, CD8+, and CD20+ cells were significantly different in neoplasms with different T. The density of CD8+ and CD20+ lymphocytes was lower in patients with nodal metastasis and higher stage.CONCLUSION: The density of tumor-associated lymphocytes can anticipate the disease progression in patients with colorectal cancer, and the density of TAL influences the control of tumour progression

A Rare Case of Soft Tissue Erdheim Chester Disease: Diagnostic Dilemma and Management

BACKGROUND: Erdheim Chester disease (ECD) is a rare form of non-Langerhans histiocytosis that still presents a diagnostic and clinical dilemma. CASE PRESENTATION: We present a rare case of ECD, young 31 male with atypical localisation and soft tissue presentation and no bone involvement. He started clinical investigations due to subcutaneous tumour mass in the lumbar spine that caused severe back pain. Skin biopsy revealed ECD with Immunohistochemistry CD68+, CD10+, CD11c+, vimentin+, S100A4+. Activating BRAFV600E mutation was positive from the tumour tissue. The patient was referred to the haematology department. PET CT was performed for initial disease staging. Treatment was started with corticosteroids (methylprednisolone 0.5 mg/kg per day), and after 7 days, a significant clinical improvement was noticed in terms of pain disappearance with no need for pain killers. After two weeks, treatment with interferon Alfa (IFN-α) was started in a dose of 3 million units 3 times per week. After 4 months of interim treatment PET, CT revealed a significant reduction of the tumour mass. Therapy with IFN-α was continued, and the patient is still clinically in good condition. CONCLUSION: It can be concluded that shortening the time of diagnosis of ECD is essential in treatment outcome of this disease. Still, large studies have to confirm the best treatment of this rare condition

HER2 Positive Gastric Carcinomas and Their Clinico-Pathological Characteristics

BACKGROUND: HER2 protein expression in gastric carcinoma, in correlation with existing, acknowledged prognostic factors which include the parameters that determine the TNM stage of the disease, could become the basis for ongoing research in the field of molecular targeted and personalised therapy.AIM: To determine the expression of the HER2 protein in gastric carcinoma and to correlate the expression of a HER2 protein with clinicopathological characteristics of the disease.MATERIAL AND METHODS: The data of HER2 protein expression and the parameters of the TNM classification were obtained from the histopathological reports of the Institute of Pathology in Skopje, and for the clinical stage we used patient’s files from the University Clinic for Abdominal Surgery in Skopje.RESULTS: The analysis of the correlation of HER2 protein expression and TNM classification parameters pointed out a significant correlation between HER2 protein expression and intragastric localisation of gastric carcinoma (P = 0.005), and the tumour grade of differentiation (P = 0.034). There was also a positive correlation between HER2 protein expression pattern and positive lymph nodes in patients with gastric carcinoma (P = 0.03). The expression pattern of HER2 +++ was significantly more common registered in patients with positive lymph nodes (P = 0.03)CONCLUSION: The expression of HER2 protein could represent a biological marker with prognostic and predictive value in patients with gastric carcinoma. Considering the high mortality rate in patients with gastric carcinoma and lack of international standardised therapeutic approach, research of the role and significance of HER2 overexpression and Trastuzumab therapy may prove useful in the development of new therapeutic strategies

EBV Positive Gastric Carcinomas and Their Clinicopathological Characteristics

BACKGROUND: The understanding of the etiopathogenesis of gastric carcinoma (GC) can be a base for development of new therapeutic methods to reduce mortality and to increase survival in patients with GC. The percentage of Epstein - Barr virus (EBV) positive gastric carcinomas is uncertain, and the etiologic importance of EBV in the pathogenesis of GC has still not been elucidated. AIM: This study aimed to determine the percentage of EBV associated GC as well as to determine their clinicopathological characteristics. MATERIAL AND METHODS: The study included 80 patients with GC who were analysed for ethnicity, local growth of a tumour (T status), the presence of nodal metastases (N), the presence of distant metastases (M), stage of the disease and degree of carcinoma differentiation. For detection of EBV, immunostainings were performed on tumour tissue and the peripheral non-tumour gastric mucosa. RESULTS: Positive immunostaining with an antibody against EBV was found in 19 (23.75%) of the 80 patients with gastric carcinomas. EBV immunostainings were significantly different in patients with or without metastasis and between patients of Macedonian and Albanian ethnicity (p < 0.0001, p < 0.009, respectively). EBV immunoexpression was significantly associated with the presence of distant metastases and with patients of Albanian ethnicity. CONCLUSION: Association of EBV immunostainings with distant metastasis in patients with GC suggests the influence of EBV infection on the progression of gastric carcinoma. Due to scarce and doubtful literature data on EBV associated GC, further studies are necessary to determine the role of EBV regarding aetiology, treatment and prognosis in patients with EBV associated gastric carcinoma