AROME-ESO Oncology Consensus Conference: access to cancer care innovations in countries with limited resources. Association of Radiotherapy and Oncology of the Mediterranean Area (AROME-Paris) and European School of Oncology (ESO - Milan).

PURPOSE: Cancer is a leading cause of mortality worldwide. Its incidence is still increasing, particularly in developing countries. Recent progresses further strengthen the differences between low/middle and high-income countries. This situation calls for joint action to reduce inequities in cancer outcomes among the patients. The Association of Radiotherapy and Oncology of the Mediterranean Area (AROME) and the European School of Oncology (ESO), have initiated joint conferences devoted to access to innovations in oncology in the Mediterranean area. The heterogeneity of the economic, political and cultural situations of the different participating countries, offers the opportunity to develop consensus conference. METHODS: Cancer prevention and treatment strategies were discussed according to existing international guidelines. The Scientific committee prepared 111 questions with an objective to prioritize the access to treatments and innovations in low/middle-income Mediterranean countries. The results from the votes of 65 oncology experts, coming from 16 countries and 33 institutions have been analysed and access priorities classified accordingly. RESULTS: Ninety six percent of the proposed general recommendations concerning national health care strategies, oncology education, and treatment organization were considered to be high priorities. Regarding access to systemic treatments, 41% of the drugs without validated predictive markers and 53% of those with validated predictive markers were considered to be 1st level priority. Only 4 biological tests were considered to be 1st level priority to access to innovation. CONCLUSIONS: AROME-ESO consensus offers to cancer specialists from developing countries a basis for discussion with health authorities and payers on the prioritization of access to innovations in cancer care.

Interlaboratory concordance in HER2 testing: Results of a Serbian ring-study.

PURPOSE: The purpose of this study was to assess the immunohistochemistry and chromogenic in situ hybridization (CISH) inter-laboratory consensus between national pathology laboratories in Serbia. METHODS: This study was conducted between 2013 and 2016. In 2013, HER2 results were evaluated using two sets of four different breast cancer specimens in five laboratories. A total of 20 immunohistochemistry and 20 CISH cases were tested. In 2014, there were 6 testing rounds, and a total of 24 specimens were analyzed, whereas in 2015 and 2016, seven testing rounds were conducted, with four additional cases (i.e. a total of 28 specimens). In 2014, 2015 and 2016, all institutions performed immunohistochemical analysis only. RESULTS: We found discrepan¬cies in HER2 immunohistochemical (IHC) results in all four surveys. IHC testing resulted in diagnostic discordance between participating centers in two (2/17) cases in 2013, two (2/24) in 2014, four (4/27) cases in 2015 and three cases (3/27) in 2016. The overall agreement among the centers was 79%, 85.5%, 83.5% and 89.4%, respectively. For CISH analyses, the results for 16 (84.2%) of 19 samples were consistent for all participants. Three results were found to be discordant, indicating a misdiagnosis rate of 15.8%. In all the discrepant cases, interinstitutional discordances were related to technical and evaluation issues. CONCLUSIONS: Our study highlights the difficulty encountered during HER2 testing using immunohistochemistry and CISH. This also emphasizes the need for rigorous quality control procedures for specimen preparation and analysis.

Locally advanced rectal cancers with simultaneous occurrence of KRAS mutation and high VEGF expression show invasive characteristics.

In this study, we investigated the mutation status of KRAS gene in pretherapeutic and preoperative biopsies in 63 specimens of locally advanced rectal cancers in order to evaluate its potential predictive and/or prognostic role. Regions of interest of KRAS exon 2 were amplified and visualized on 2% agarose gel. Obtained PCR products were subjected to direct sequencing. KRAS mutations were detected in 35% of patients, 91% of which were located in codon 12 and 9% in codon 13. In general, KRAS mutation status did not affect the response to neoadjuvant chemoradiotherapy (CRT). However, patients harboring mutated KRAS gene, simultaneously with high vascular endothelial growth factor (VEGF) expression, exhibited a worse response to CRT (p=0.030), a more frequent appearance of local recurrences and distant metastasis (p=0.003), and shorter overall survival (p=0.001) compared to all others. On the contrary, patients with GGT>GCT KRAS mutation exhibited a significantly better response to CRT than those with any other type of KRAS mutation (p=0.017). Moreover, the presence of GGT>GCT mutation was associated with low VEGF and Ki67 expression (p=0.012 in both cases), parameters related to less aggressiveness of the disease. Our results suggest that KRAS mutation status could have some predictive and prognostic importance in rectal cancer when analyzed together with other parameters, such as VEGF and Ki67 expression. In addition, it seems that not only the presence but the type of KRAS mutation is important for examining its impact on CRT response.

Successful treatment with cladribine of Erdheim-Chester disease with orbital and central nervous system involvement developing after treatment of Langerhans cell histiocytosis.

INTRODUCTION: Erdheim-Chester disease (ECD) is a rare, systemic form of non-Langerhans cell histiocytosis of the juvenile xanthogranuloma family with characteristic bilateral symmetrical long bone osteosclerosis, associated with xanthogranulomatous extra skeletal organ involvement. In ECD, central nervous system (CNS) and orbital lesions are frequent, and more than half of ECD pa tients carry the V600E mutation of the protooncogene BRAF. The synchronous or metachronous development of ECD and Langerhans cell histiocytosis (LCH) in the same patients is rare, and the possible connection between them is still obscure. Cladribine is a purine substrate analogue that is toxic to lymphocytes and monocytes with good hematoencephalic penetration. CASE REPORT: We presented a 23-year-old man successfully treated with cladribine due to BRAF V600E-mutation-negative ECD with bilateral orbital and CNS involvement ECD developed metachronously, 6 years after chemotherapy for multisystem LCH with complete disease remission and remaining central diabetes insipidus. During ECD treatment, the patient received 5 single-agent chemotherapy courses of cladribine (5 mg/m2 for 5 consecutive days every 4 weeks), with a reduction in dose to 4 mg/m2 in a fifth course, delayed due to severe neutropenia and thoracic dermatomal herpes zoster infection following the fourth course. Radiologic signs of systemic and CNS disease started to resolve 3 months after the end of chemotherapy, and CNS lesions completely resolved within 2 years after the treatment After 12-year follow-up, there was no recurrence or appearance of new systemic or CNS xanthogranulomatous lesions or second malignancies. CONCLUSION: In accordance with our findings and recommendations provided by other authors, cladribine can be considered an effective alternative treatment for ECD, especially with CNS involvement and BRAF V600E-mutation-negative status, when interferon-alpha as the first-line therapy fails.

Giant esophageal fibrovascular polyp with clinical behaviour of inflammatory pseudotumor: a case report and the literature review.

INTRODUCTION: Esophageal fibrovascular polyps are rare, benign, intraluminal, submucosal tumor-like lesions, characterized by pedunculated masses which can demonstrate enormous growth. The most frequent symptoms are dysphagia, vomiting and weight loss. Fibrovascular polyps with long stalks can regurgitate into the airways and cause asphyxia. Esophageal inflammatory pseudotumor is extremely rare lesion accompanied with various systemic manifestations as fever, anemia and thrombocytosis. CASE REPORT: We presented a 29-year-old man complaining of a long-lasting fever and dysphagia. He was found to have huge pedunculated submucosal tumor of esophagus, surgically completely resected. Histopathological examination showed that this giant tumor, 24 x 9 x 6 cm, was a fibrovascular polyp. The postoperative course was uneventful. The preoperative fever, anemia and thrombocytosis disappeared and did not recur in the postoperative course. CONCLUSION: We reported a patient with giant esophageal pedunculated tumor with clinical manifestations of inflammatory pseudotumor and histopathological picture of fibrovascular polyp, that we have not found described in the literature before.

Hypermethylation of p15 gene in diffuse - large B-cell lymphoma: association with less aggressiveness of the disease.

In this study, methylation-specific polymerase chain reaction was used to investigate the potential prognostic significance of the methylation status of p15, p16, MGMT, and DAPK genes in 51 specimens of diffuse large B-cell lymphoma (DLBCL). Hypermethylation of p15 gene was significantly more prevalent in patients without relapse (p = 0.001) and there was a trend toward more frequent presence of p15 methylation in patients without death outcome within 5-year follow-up period (p = 0.086) Also, there was a trend toward accumulation of p15 methylation with favorable clinicopathological parameters including: age ≤ 60 years (p = 0.091), normal levels of lactate dehydrogenase (p = 0.090), Eastern Cooperative Oncology Group performance status < 2 (p = 0.095), and low/intermediate low International Prognostic Index (p = 0.076). In the female group and group of the patients without bulky tumor mass, treated with chemotherapeutic regimens including rituximab, methylation of p15 was significantly related to longer overall survival (p = 0.036 and 0.027, respectively). Our results suggest that promoter methylation of p15 gene could have prognostic value in DLBCL patients treated with rituximab when used in combination with gender and tumor size.

Comparsion the histopathological findings after cervical biopsy and excisional procedures.

INTRODUCTION: A definitive diagnosis of cervical intraepithelial neoplasia (CIN) is confirmed after histopathological (HP) examination of the tissue obtained through the biopsy. The aim of this study was to compare histopathological results obtained with punch biopsy and results obtained through one of the excisional techniques. MATERIAL AND METHODS: We analysed histology results of 130 patients referred to our institution with abnormal smear. Punch biopsy was performed after colposcopic examination in all patients before one of the excision methods. Excision methods performed were: large loop excision of transformation zone (LLETZ), radio-frequency knife conisation or cold knife conisation. Based on the histopathological examination of the punch biopsy specimen or excisional specimen diagnosis of CIN was established. RESULTS: CIN and invasive cancer were the most common diagnoses in the 31-40 age group at 45.4% (59/130). Discrepancies in the histological diagnosis between punch biopsy and excisional biopsy was identified in 58.5% (76/130) of the patients. In 6% of the of the cases the biopsy did not detect an invasive carcinoma. CONCLUSION: The most frequent discrepancies between punch biopsy and excisional biopsy were in the group of patients with a higher grade cervical dysplasia. Mild dysplastic changes diagnosed through punch biopsy, require a more conservative approach, as the majority of this group had negative specimens on the cone after excision, especially in the younger population. It is advisable that the patients above 30 years of age and a higher grade dysplasia in the biopsy specimen, should undergo one of the excisional techniques as a diagnostic/therapeutic method of treatment.

[Diagnostic relevance of fine needle aspiration cytology in nodular thyroid lesions].

BACKGROUND/AIM: Fine needle aspiration cytology (FNAC) of the thyroid gland has been used as an initial investigative procedure of thyroid nodule(s) in the Department of Pathology at the Institute of Oncology of Vojvodina for more than 20 years. This procedure is rapid, inexpensive and technologically simple, yet it has found only limited, albeit increasing acceptance in medical practice in Serbia. The aim of the study was to evaluate our FNAC findings by correlating cytological results with histological diagnosis and to define the sensitivity, diagnostic accuracy and positive predictive value of FNAC. METHODS: A total of 266 patients with thyroid swellings were aspirated in one year investigated period at our Institute out of whom 69 underwent surgery between May 2008 and May 2009. The cytological results correlated with clinical features, ultrasound investigations (US) and subsequent histopathological examination of the resected tissue. RESULTS: By the use of cytology we found out thyroid carcinoma in 10 patients, and by histopathological examination in 12. We obtained 83% sensitivity, 100% specificity and 97% of diagnostic accuracy of FNAC. CONCLUSION: The obtained results confirm the importance of FNAC in preoperative assesment of thyroid nodule.

[Expression of cytokeratins 5/6 and cytokeratin 17 in invasive breast carcinoma].

BACKGROUND/AIM: Cytokeratins (CK) 5/6 and 17, myoepithelial markers, are also expressed in a proportion of breast carcinomas. Breast carcinomas expressing basal epithelium cytokeratins constitute a tumor subgroup that shows common but heterogeneous morphological, genetical, and immunophenotypical features and is associated with poor clinical outcome. The aim of this study was to determine the incidence of basal expression of cytokines CK5/6 and CK17 in the tested samples of ductal invasive breast cancers, as well as to test the presence of a correlation of tumor expression of basal cytokines and clinicopathological prognostic factors: age, the level of histological differentiation, hormone receptor status, HER2 (human epidermal growth factor receptor 2) protein expresion and HER2 gene amplification in tumorous tissue. METHODS: Immunohistochemistry (IHC) was used to evaluate the CK5/6 and CK17 status of 121 ductal invasive breast cancers. The results thus obtained were compared with clinicopathological characteristics. RESULTS: From the 117 analyzed tumor specimens, 22% and 30% were immunohistochemically positive for CK5/6 and CK17, respectively. Basal cytokeratins showed significant inverse relationship with estrogen and progesterone receptor status and HER2 protein expression. CK5/6 and CK17 immunoreactivities were directly associated with triple-negative phenotype and higher histological grade. CONCLUSION: Our findings are similar to reports that tumours expression of basal cytokeratins are correlated with adverse pathological parameters. Given the limited number of emerging therapeutic targets in these tumors, routine IHC identification of basal-like subtype as a poor prognostic group of breast cancer could be based on the expression of basal CKs.

Colorectal carcinoma under microscopy: patohistology or much more?

Today the role of pathologists is increasingly focused on finding more accurate prognostic and predictive parameters that will be necessary for targeted treatment of patients. Improving understanding of colorectal carcinogenesis allow us to consider incorporation of these new knowledges in molecular classification of colorectal cancer. There are different ways of molecular classification, but most of them are based on: 1. type of genetic instability; 2. methylator phenotype and 3. single molecular events such are KRAS and BRAF mutations. This review considers a new molecular classification of colorectal carcinoma proposed by J. Jass in 2007 which is based on the correlation of molecular and morphological features. We would also like to point out to the new role of pathologists in the era of personalized medicine in diagnosis and prognosis of colorectal carcinomas as well as in selection of patients for some modalities of targeted therapy.

[Human epidermal growth factor receptor 2 testing in breast cancer].

INTRODUCTION: Testing for human epidermal growth factor receptor-2 in breast cancer at the time of primary diagnosis is now the standard of care. Positivity for epidermal growth factor receptor-2 in breast cancer is a prognostic factor regarding tumor aggressiveness and a predictive factor for response to Herceptin. Accurate assessment is essential to ensure that all patients who may benefit from Herceptin are correctly identified. ASSAY METHOD: The principal testing methods used for determination of epidermal growth factor receptor-2 status are immunohistochemistry for protein overexpression and in situ hybridization using either fluorescence or a chromogen. Immunohistochemical testing method allows identification of epidermal growth factor receptor-2 positive patients (3+) who may benefit from Herceptin therapy, whereas negative patients (0/1+) can be excluded. A proportion of specimens defined as equivocal by immunohistochemistry (2+) must be retested by in situ hybridization to determine their status. Chromogen in situ hybridization is a method for determination of gene amplification using a peroxidase-based chromogenic reaction, which can be viewed using a conventional bright field microscope and it determines the actual degree of gene amplification. Various factors can affect the results achieved with these assays, including the assay antibody/probe, the methodology and the experience of personnel. Many countries implemented national testing guidelines in an attempt to standardize testing procedures and make results more accurate. CONCLUSION: The key point underlined by this review is that whatever method is used to test HER2 status, the technology must be validated first, and there must be regular internal and external quality assurance procedures.

Correlation of HER-2/neu protein overexpression with other prognostic and predictive factors in invasive ductal breast cancer.

BACKGROUND: The objective of our study was to investigate the association between Her-2/neu status and other clinicopathological characteristics of ductal breast carcinoma. PATIENTS AND METHODS: A total of 120 cases of breast carcinoma were included in this study. The immunohistochemical staining for HER-2/neu, hormone receptors, p53 and Ki-67 were evaluated. RESULTS: HER-2/neu protein overexpression was present in 4 out of 63 T1 lesions, in 13 out of 44 T2 lesions, in 3 out of 7 T3 lesions, and in 3 out of 6 T4 lesions. Protein overexpression was found in 10 out of 21 grade III tumors and 13 out of 72 grade II tumors. Overexpression was not detected in grade I tumors. Of the 23 Her-2/Neu-positive cases, ER- and PR-negative status was detected in 61% and 69%, respectively. Her-2 protein overexpression was seen in 23 out of 93 high Ki-67 tumors, whereas overexpression was not detected in low Ki-67 cases. CONCLUSION: Statistically significant correlation was found between HER-2/neu protein overexpression and large tumour size, high histological grade, ER and PR negativity, and high Ki-67 proliferative index.

Prognostic value of thymidylate synthase, epidermal growth factor receptors and vascular endothelial growth factor in patients with stage III colon cancer.

PURPOSE: We studied the prognostic value of thymidylate synthase (TS), Epidermal Growth Factor Receptors (EGFR) and Vascular Endothelial Growth Factor (VEGF) expression in primary colon cancer (CC). PATIENTS AND METHODS: Those three markers were immunohistochemically assessed on tumour sections from 100 patients with CC Dukes C. All patients received the same adjuvant chemotherapy with FU/Leukovocin according to Mayo protocol. Considering the clinical course they were classified in two groups: bad in which all patients progressed and good in which neither progressed during the five year follow up period. RESULTS: TS, EGFR and VEGF were an independent prognostic factor for time to progression (TTP) and overall survival (OS). Findings of at least two maximum expressed investigated markers, significantly increases the risk of progression which influences shorter five year survival, and the single maximum expression does not necessarily have to be a bad prognostic sign. CONCLUSION: Highest expression of TS, EGFR and VEGF carries prognostic significance with respect to TTP and OS for patients with Dukes C colon cancer.

[The influence of aging on pathologic and immunobiologic parameters of invasive ductal breast carcinoma].

BACKGROUND/AIM: Most human cancers, including breast one, increase in frequency with aging. The aim of this study was to explore the hypothesis that aging also alters breast cancer biology. METHODS: The study included 120 women with primary invasive ductal carcinoma of the breast. We correlated the patients age and diagnosis with the commonly used clinical, pathological factors and newer tumor biomarkers. Immunohistochemical staining was conducted for p53, c-erbB-2, Ki-67, estrogen (ER), progesterone (PR) receptors, and angiogenesis. RESULTS: In our study, the patients with axillary lymph node metastases and negative steroid hormone receptors (ER and PR) were significantly younger than the patients with nodal involvement and positive hormone receptors. There was also a significant association between the patients age, diagnosis and angiogenesis. No association was found between the patients age and tumour size, histological grade, p53, c-erbB-2, and Ki-67. CONCLUSION: The results of our study supported only partially the hypothesis that the breast cancer biology is significantly affected by a patient's age.

A human origin type II strain of Toxoplasma gondii causing severe encephalitis in mice.

Despite its capacity for sexual reproduction and global distribution, Toxoplasma gondii has a highly clonal structure, with the majority of isolates belonging to three distinct clonal types. Congenital toxoplasmosis has been associated with type I and type II strains. We here present the first characterization of a T. gondii strain (BGD1) from South-East Europe, isolated from the umbilical blood of a 24-week-old fetus in Serbia. Genotyping, performed by PCR-RFLP using a set of nested PCR markers (5'SAG2, 3'SAG2, BTUB, SAG3, and GRA6), showed that the BGD1 strain possessed a type II genotype. The cytokine patterns in Swiss-Webster mice inoculated with brain cysts of BGD1 and the prototype type II ME49 strain were similar until 180 days post-infection, with highly elevated IFN-gamma, IL-12 and IL-10 by d7 and decreasing thereafter. While both strains induced pneumonia and hepatitis in acute infection (d14), chronic infection (d56) was characterized, in addition to hepatitis, by severe meningoencephalitis, associated with numerous brain cysts. Thus, the BGD1 strain of T. gondii has type II genotypic and immunologic characteristics, but unlike other type II strains of human origin, induces severe encephalitis, making it an alternative to the sheep-derived ME49 strain for experimental models of infection.

[Occurrence of autoimmune chronic hepatitis in siblings].

BACKGROUND: Autoimmune hepatitis is a chronic inflamatory disease of the liver of unknown etiology, characterized by the loss of tolerance against hepatic tissue, leading to the destruction of hepatic parenchyma. It predominantly affects females, and rarely occurs in the same family. CASE REPORT: In this paper we presented brother and sister with autoimmune hepatitis according to the criteria of the International Autoimmune Hepatitis Group. CONCLUSION: Because of a possible genetic predisposition to autoimmune hepatitis, the occurrence of the disease in a family member suggested the need to examine other family members.