Identification of the c.829_832delAATA Deletion Variants in the BRCA1 Gene Associated with Hereditary Breast/Ovarian Cancer - Case Report.

Determination of the BRCA1/BRCA2 mutation status in patients with breast and/or ovarian cancer is commonly performed using various molecular techniques. The use of targeted PCR-based tests only may not be sufficient, as not all possible variants are investigated. In the present study, we used next-generation sequencing (NGS) techniques to identify novel pathogenic variants in BRCA1 and BRCA2. In this study, material (blood and FFPE) collected from a 67-year-old patient with ovarian cancer was used. The presence of hereditary mutations characteristic for the Polish population was examined using Sanger sequencing. BRCA1 and BRCA2 gene exons were amplified using the Devyser BRCA kit and sequenced on the Miniseq. No germline mutations characteristic for the Polish population were detected. However, 12 single nucleotide variants and 2 indels were identified. We found a new deleterious mutation of gene BRCA1 (c.829_832delAATA). To our knowledge, this mutation has not been reported yet in the Polish population and elsewhere. The use of the NGS technique increases the possibility of detecting mutational changes in patients with ovarian and/or breast cancer. Quick determination of pathogenic variants is important to facilitate specific therapy, in addition to the identification of familial predisposition to cancer.

HER2 gene assessment in liquid biopsy of gastric and esophagogastric junction cancer patients qualified for surgery.

BACKGROUND: Amplification of HER2 gene (ERBB2) and overexpression of HER2 protein on cancer cells are found in 10-26% of gastric cancer (GC) and esophagogastric junction cancer (EGJC). Gene copy number variation (CNV) could be detected in these patients in liquid biopsy and in cancer cells. METHODS: We analysed HER2 gene CNV used qPCR method in 87 sera collected from GC and EGJC patients before surgical treatment and in 40 sera obtained from healthy donors. HER2 gene CNV was also assessed in formalin-fixed paraffin-embedded (FFPE) tumor tissue. Furthermore, we assessed the number of HER2 gene copies and HER2 expression in cancer cells using the fluorescent in situ hybridization method (FISH) and immunohistochemistry (IHC). RESULTS: We found that the HER2 gene copy number in liquid biopsy was higher in GC and EGJC patients compared to healthy people (p = 0.01). Moreover, EGJC patients had higher number of HER2 gene copies than healthy donors (p = 0.0016). HER2 CNV examination could distinguish healthy individuals and patients with gastric or esophagogastric junction cancers with sensitivity and specificity of 58% and 98% (AUC = 0.707, 95% CI 0.593-0.821, p = 0.004). We found that patients with a high copy number of the HER2 gene in the tumor tissue assessed by qPCR (but not by FISH) have significantly more often a high number of HER2 gene copies in liquid biopsy (p = 0.04). CONCLUSIONS: We suggested that HER2 testing in liquid biopsy could be used as an auxiliary method to analysis of HER2 status in tumor tissue in gastric or esophagogastric junction cancers.

Perfusion CT - A novel quantitative and qualitative imaging biomarker in gastric cancer.

OBJECTIVES: The aim of this research was to examine whether Perfusion Computed Tomography (P-CT) can qualitatively and quantitatively help detect gastric cancer neoangiogenesis in vivo as well as treatment response evaluation. We attempted to explore which P-CT parameters are best used in neoangiogenesis and neoadjuvant therapy for most effective evaluation. We also tried to recognize a positive prediction value of P-CT in early responders and non-responders patients identification. MATERIALS AND METHODS: Twenty-four patients with positive biopsy results and/or clinically proven gastric cancer were enrolled in the P-CT exam. Patients were qualified for systemic treatment (16 patients received chemotherapy and 8 patients received radiochemotherapy). The baseline Perfusion-CT exam and after neoadjuvant treatment Perfusion-CT exam were conducted using a 64-row GE tomograph based on a deconvolution model in first-pass protocol perfusion. The P-CT examined the following parameters: Blood Flow (BF), Blood Volume (BV), Mean Transit Time (MTT) and Permeability Surface (PS). Positive clinical response to neoadjuvant treatment (CHT and RCT) was defined as tumor size reduction 25% or more. RESULTS: Tumor dimension reduction after neoadjuvant therapy was significantly correlated with the BF and the PS. Neoadjuvant therapy was more effective for patients with higher output BF and PS values. We did not register a significant relationship between BV and MTT parameters and tumor dimension reduction. Patients with a positive treatment response showed a decrease in BF, BV and PS perfusion parameters with an increase in MTT. CONCLUSIONS: P-CT examination allows a noninvasive neoangiogenesis assessment in vivo, leading to early identification of responding and non-responding patients. As a standard procedure, a full evaluation of treatment response should include a P-CT exam assessing neoangiogenesis.

Cat scratch disease in 9-year-old patient - a case report.

Cat scratch disease (CSD) - bartonellosis, is zoonosis caused by the intracellular gram negativebacterium Bartonellahenselae or Bartonellaquintana. The pathogens of this disease enter the human body usually as a consequence of a bite or scratch by young cats which are the natural source of such bacteria. The illness proceeds asymptomatically or with topical symptoms of infection such as a lump, spot or blister. Within 14 days a high fever and topical lymphadenopathy are observed. Lymph nodes are sore and start suppurating. In half of patients, these symptoms may resemble malignancy, and in single cases there are symptoms associated with the musculoskeletal system, such as: osteitis, arthitis and myositis. In paper presented case of 9 year-old girl patients, treated in Oral Surgery Unit due to odema and lymphadenopathy in right submandibular space. Primary surgical treatment of deciduous teeth was conducted without recovery. In few months follow-up, biopsy of lymph node of submandibular group was taken and provisional diagnosis of cat scratch disease was set. Patient was referred to the Infectious Diseases Unit where serological test confirmed cat scratch disease, and pharmacological treatment was conducted with success and recovery of young patient.

Microsatellite polymorphism in the P1 promoter region of the IGF­1 gene is associated with endometrial cancer.

Endometrial carcinoma (EC) is the most common type of gynecological malignancy. Studies have demonstrated that the insulin growth factor (IGF) pathway is implicated in the development of endometrial tumors and that the serum levels of IGF­1 are affected by estrogen. Most EC cells with high microsatellite instability (MSI­H) accumulate mutations at a microsatellite sequence in the IGF­1 gene. The present study investigated the CA repeat polymorphism in the P1 promoter region of the IGF­1 gene among Caucasian females with endometrial hyperplasia, EC and healthy control subjects, whose blood serum and surgical tissue specimens were analyzed. Differences or correlations between the analyzed parameters [serum levels of IGF-1 and IGF binding protein (IGFBP)­1 and IGFBP­3 as well as estrogens among the polymorphisms] were verified using the χ2, Mann-Whitney U, Kruskal-Wallis or Spearman's rank correlation tests. A PCR amplification and DNA sequencing analysis was used for identification of (CA)n repeats in the P1 region of IGF­1. ELISA was used to determine the blood serum levels of IGF­1, IGFBP­1, IGFBP­3 and estrogens. Furthermore, IGF-1 was assessed in endometrial tissues by immunohistochemical analysis. The present study indicated no statistically significant differences between serum levels of IGF­1, IGFBP­1, IGFBP­3 and estrone, estriol and estradiol in the control and study groups. A significant correlation was identified between the IGF-1 levels and estrone levels in the MSI-H polymorphism (r=-0.41, P=0.012) as well as a highly negative correlation between IGF-1 levels and the estradiol levels in the MSI-H polymorphism (r=-0.6, P=0.002). Genotypes without the 19 CA allele were predominantly found in EC. Furthermore, statistical analysis indicated that the number of IGF-1-expressing cells was significantly elevated in MSI-H type 18-20 (P=0.0072), MSI-L type 19-20 (P=0.025) and microsatellite-stable MSS type 19-19 (P=0.024) compared with those in the MSI-H 20-20 genotype. The present study suggested that it is rather likely that the polymorphisms in the IGF-1 promoter are associated with EC in Caucasian females with regard to its development. In the present study, polymorphisms of the IGF-1 promoter may have been introduced during the genesis of EC and contributed to it by leading to aberrant expression of IGF-1.

The influence of a thyroxine supplemented diet on selected hepatic redox equilibrium markers, liver morphology and the serum lipid profile in rats treated with doxorubicin.

INTRODUCTION: Cytotoxicity of doxorubicin (DOX) - an anticancer drug, mostly results from reactive oxygen species (ROS) generation. Some enzymes catalyzing this process and enzymes of antioxidant defense are regulated by iodothyronine hormones. Thus, disorders in iodothyronine hormone status may affect doxorubicin-induced redox imbalance and anabolic/catabolic disorders. The aim of this study was to evaluate the influence of doxorubicin and thyroxine (T4) associated treatment on liver morphology, markers of oxidative stress and plasma lipid parameters. MATERIALS AND METHODS: Rats were intraperitoneally treated with doxorubicin (1.5 mg/kg) once a week for ten weeks. Thyroxine was simultaneously given in drinking water (0.2 or 2.0 mg/l) for 14 weeks. RESULTS: There were higher hepatic level of malonyldialdehyde (MDA) of all tested groups and at the same time in rats treated with DOX plus T4 lower concentrations of total glutathione compared to controls were observed. Morphology of liver did not show any features of necrosis or steatosis but a decrease of glycogen content in T4+DOX groups compared to DOX treatment was observed. The concomitant administration of a lower dose of thyroxine and doxorubicin decreased triglycerides (TG) and increased LDL level compared to the DOX group. DISCUSSION: Thyroxin supplementation caused redox equilibrium disorders and oxidative stress in liver of rats receiving DOX. The study revealed the normalizing influence of thyroxin on glycogen deposits that were observed after doxorubicin treatment. Apart from an adverse impact of thyroxine administration on LDL in rats treated with doxorubicin, a beneficial effect of lower dose of thyroxine on serum TG level was revealed.

Interaction of lymphocytes and thyrocytes in Graves' disease and nonautoimmune thyroid diseases in immunohistochemical and ultrastructural investigations.

BACKGROUND: Graves' disease is the archetype for organ-specific autoimmune disorders. It is very important for our understanding of the mechanisms responsible for progression of autoimmunity. The aim of this study was to present interactions of lymphocytes and thyrocytes in the thyroid tissue in Graves' disease and nonautoimmune thyroid diseases. METHODS: The study involved 30 children with Graves' disease, 30 children with nodular goiter, 30 with simple goiter and 30 healthy children. After thyroidectomy, T cells were detected in the thyroid specimens by CD3, CD4, CD8 antibodies, B cells by CD79alpha antibodies and the antigen-presenting dendritic cells with CD1a antibodies (DakoCytomation) and were examined in the EM 900 Zeiss Germany Electron Microscope. RESULTS: The most enhanced immune reaction was observed in the thyroid from children with Graves' disease. The cells of the immune system infiltrated the thyroid follicles and interfollicular compartments; they also formed lymph follicles. CONCLUSION: The immune reaction in Graves' disease and migration of lymphocytes T and B between thyrocytes results in the thickening of the basal membrane of the thyroid follicle. No cytotoxic effect of T cytotoxic/suppressor CD8+ cells on thyrocytes was observed in Graves' disease, while a mild cytotoxic effect was observed in non-autoimmune thyroid disease.