Savelina L. Popovska, Ivan N. Ivanov and Akishi Ooi
Triple-negative breast cancer (TNBC) is a subtype of breast cancer which is clinically negative for expression of steroid hormones and HER2 protein. According to DNA microarray profiling, breast cancer was divided into several molecular subgroups, including Luminal A and Luminal B; HER2 overexpressing; normal breast- like and basal-like subtype. TNBC comprises tumours that express heterogeneous molecular and immunohistochemical phenotype, thus determining different prognosis. The majority of TNBC carry the “basal-like” molecular profile on gene expression arrays. Basal-like carcinomas of the breast associated with poor prognosis often correlate with expression of Caveolin-1. The study aimed to investigate the expression of caveolin-1 in the tumour cells and in the stromal component of the tumour among the group of TNBC and compare it to the expression of caveolin-1 in a control group with non-TN breast cancer. Whole tissue sections were used. Formalin fixed, paraffin embedded tissue materials from 101 patients diagnosed with invasive breast cancer during the period 2004-2007 were investigated in a retrospective study. A multistep approach was used to separate the different subtypes of breast cancer. During the first step the breast tumours were separated according to their ER, PgR, HER 2 and proliferative activity, using the Ki-67 index. The triple- negative tumours were additionally tested for EGFR and CK5/6. The basal breast cancer group was finally subdivided into basal and baso-luminal according to the type of expression of basal cytokeratins. caveolin-1 expression was examined in the tumour parenchyma and stroma. According to our results, caveolin-1 expression in breast cancer was significantly associated with basal biomarkers expression (basal and baso-luminal type of BC) χ2= 33.4; p<0.0001. Caveolin-1 can be used as a potential marker to examine the presence of basal phenotype in breast cancer.
Valeri Y. Andreev, Danail B. Petrov, Ivan N. Ivanov and Yavor Y. Ivanov
Deciding on a treatment approach for early stage lung cancer (0-1) is sometimes difficult because of uncertainties regarding the depth of tumour invasion and its margins. Even with advanced technology, such as endobronchial ultrasound and autofluorescence bronchoscopy, it is often difficult to be precise. In this currently discussed case, treatment of a 61-year-old female patient with early stage IA lung cancer could not proceed for such reasons. Fortunately, timely surgical intervention allowed preservation of lung function and the patient is now under close surveillance.
Ivan N. Ivanov, Savelina L. Popovska, Emilian A. Ivanov, Sergey D. Iliev and Pencho T. Tonchev
The phenomenon “vessels within vessels” was initially described by Merchant S et al. as arteries found free-floating inside the lumen of veins. We have described another version of the “vessels within vessels”complex, composed of arteries found inside the lumen of lymphatic vessels. The purpose of the present study was to describe the structure of the lympho-arterial vascular complex in a breast tissue specimen from a male subject using three- dimensional tissue reconstruction. A histological specimen from a 64 year-old male subject diagnosed with gynecomastia was used. The tissue sample was sectioned in a multi-step manner. An overall of approximately 150pm thick tissue material was sampled. Immunostaining with anti-CD34 antibody and anti-podoplanin antibody was performed. Three-dimensional reconstruction of the vessel within the vessel structure was performed with “Reconstruct” software When the reconstruction of the breast parenchyma was revealed as a 3D image, it became apparent that the arterial vessel was situated inside the lymphatic vessel and could be followed along the entire length of the vascular segment studied. We have proved that these vascular complexes are not artificial phenomenon and do exist. The function of the vascular complexes is still uncertain, and is probably related to lymph propulsions in the initial collector vessels.
Savelina L. Popovska, Ivan N. Ivanov and Dorian Y. Dikov
Neoadjuvant chemotherapy is the standard of care for patients with locally advanced breast cancer (LABS). The aim of this study was to determine the effects of neoadjuvant chemotherapy on metastatic and nonmetastatic breast cancer axillary lymph nodes (ALNs). Seventy-seven patients with LABS and cytologically documented ALN metastases were treated in two prospective trials. Patients had breast surgery with level I and level II axillary dissection followed by additional chemotherapy or radiation treatment. Clinical nodal status was determined by physical examination and compared with histological and immunohistochemical findings. The regressive changes in primary tumor and ALNs were achieved using scoring systems. Objective clinical response in primary tumor was seen in 48.05% of patients. Histologic evidence of regression was noted in 46 cases (59.75%), with complete histologic regression achieved in 5 cases (6.49%). The number of cases according to the ALNs regression scoring system was as follows: 0 grade - viable metastases without effect - 21 patients (30%); 1 grade - cytotoxic effect in metastases - 20 patients (28.58%), 2 grade - micrometastasis - only3 patients (4.28%); 3 grade - no residual metastatic disease - 1 patient (1.43%); 4 grade - negative ALNs before and after treatment - 25 patients (35.71%). Nodular fibrotic area and iron-loaded macrophages suggested previous presence of tumor metastasis. There was significant relationship between histologic regression in the primary tumor and the presence of effect of cytotoxic agents on ALNs metastases (r=0.9123; p<0.00001). As chemotherapy is widely used in the treatment of breast carcinoma, pathologists should be aware of chemotherapy-induced changes in metastatic and in nonmetastatic ALNs.
Valeri Y. Andreev, Nikolay A. Yanev, Stefan K. Stanimirov, Ivan N. Ivanov, Savelina L. Popovska, Petkana A. Hristova and Yavor Y. Ivanov
Background: Autofluorescence bronchoscopy (AFB) allows a more sensitive approach to the diagnosis of premalignant and malignant endobronchial lesions than white light bronchoscopy (WLB) can do.
Aim: To assess the autofluorescence bronchoscopy and white light bronchoscopy in diagnosing malignant endobronchial lesions.
Materials and methods: The design of the study is a retrospective case-control study. Thirty-two parameters were entered into an Excel file and analysed with SPSS v. 21 for Mac book Pro. Endoscopy findings were graded in 4 options and morphological results - in 9 options according to WHO classification. The results are presented using McNemar’s test and sensitivity, specificity and positive and negative predictive values as well.
Results: Three hundred and three patients were included in the study. Lung cancer was found in 38.3% of the patients using histology and in 35.6% - using cytology. McNemar’s test for AFB finding for suspected and malignant lesions OR was 8.333 (95% CI 3.571-23.784) while for WLB OR was 0.128 (95% CI 0.045-0.299). For cytological results OR was 3.800 (95% CI 2.123-7.227) and 3.471 (95% CI 1.996-6.351), respectively. P value was <0.0001 for all tests. Sensitivity for AFB and WLB was 94.83% but specificity was 52.83% and 55.66% if histology was used. For cytology these numbers were respectively 86.11% and 84.26% for sensitivity, and 63.69% and 62.42% for specificity.
Conclusion: AFB has an advantage over WLB in diagnosing endobronchial malignant lesions. Biopsying suspicious, not only visible malignant lesions, increased diagnostic sensitivity.
Valeri Y. Andreev, Nikolay A. Yanev, Stefan K. Stanimirov, Temenuzhka V. Mircheva, Ivan N. Ivanov, Savelina Popovska, Petkana A. Hristova and Yavor Y. Ivanov
As part of a retrospective study on bronchoscopies performed at the Clinic of Pneumonology and Phthisiatry of the University Hospital – Pleven by autofluorescence bronchoscopy we found 3 cases diagnosed with carcinoma in situ. They were treated in different ways – endobronchial electrocoagulation, extraction by forceps biopsy and open surgery, but the result was the same – clinical healing. The paper presents the three clinical cases and the analysis of the treatment.
Katia S. Kovacheva, Zornica B. Kamburova, Savelina L. Popovska, Ivan N. Ivanov, Maria N. Simeonova and Petia N. Angelova
Genetic testing for BRCA 1/2 mutation is a well recognized medical management tool. Identification of healthy carriers of such mutations allows effective risk reduction procedures to be performed. There is no data reported on the founder mutations in the Bulgarian population. To evaluate the contribution of genetic factors to breast cancer (BC), we investigated the carrier state of Bulgarian women with BC for five common (according to BIC database) deleterious BRCA1/2 mutations. The list of patients diagnosed with BC between January 2011 and April 2012 was obtained from the Cancer Registry of University Hospital, Pleven. Eighty-two women with BC were interviewed and a pedigree was constructed of each of them. The patients were classified into seven categories, according to personal, disease and family history. Based on the preliminary prepared selection criteria and the personal family history, we defined a target group of 33 Bulgarian women with BC. They were screened for five deleterious mutations: 5382insC in BRCA1 and 6174delT, 6079del4, 8138del5, 5946delCT in BRCA2, by DNA sequencing. The genetic analysis detected none of the tested mutations. Two polymorphic variants were found in BRCA2 gene: c.5744C>T (rs4987117, SNP database) in exonl 1E in one patient and c.7806-14T>C (rs9534262, SNP database) in exonl7 in 22 patients. In conclusion, without basic information on the founder mutations in the population, the genetic screening for the specific mutations in a small group of tested patients is ineffective.
Katia S. Kovacheva, Zornitsa B. Kamburova, Savelina L. Popovska, Dobromir D. Dimitrov, Ivan N. Ivanov, Maria N. Simeonova and Tashko S. Deliyski
Detection of mutations in breast cancer 1 (BRCA1) and breast cancer 2 (BRCA2) gene is an effective method of early diagnosis and prevention of breast cancer (BC). The mutational spectrum of both genes in Bulgarian population has not been studied in depth. The aim of this study was to investigate the prevalence of five deleterious BRCA1/2 point mutations in high-risk BC women, selected according to the National Comprehensive Cancer Network (NCCN) Guidelines including early age of onset, triple-negative BC and family history of breast or ovarian cancer. The prevalence of two BRCA1 mutations (C61G and 5382insC) and three BRCA2 mutations (6079del4, 9326insA and 9908delA) was evaluated in 80 females with BC, obtained from the Cancer Registry of University Hospital - Pleven. Genetic testing was performed by direct DNA sequencing. One deleterious mutation (5382insC in exon20 in BRCA1) was been found in two patients (2.5%). Both women were diagnosed with BC before age 45. The prevalence of BRCA mutations established in our study was lower than the one found in another preliminary study on Bulgarian population. We concluded that this discrepancy was due to the genetic heterogeneity of the population and the specific mutational spectrum of the BC patients from the Pleven region.
Dancho P. Dekov, Ivan N. Ivanov, Sergey D. Kostadinov, Savelina L. Popovska, Petko I. Lisaev and Plamen D. Dorovski
The study is a statistical analysis of the dynamics and structure of forensic autopsies performed on dead bodies and body remains at University Hospital “Dr. Georgi Stranski” in Pleven. The study was based on forensic autopsy records database and forensic autopsy reports for the period 2009-2013. A total of 976 cases, including forensic autopsies, exhumations, examination of bone remains and inspection of dead bodies at the site of death (without following forensic autopsy), were analyzed. The studied data was coded and processed with Microsoft Excel software. A detailed data analysis of the number of autopsies, frequency of death by cause, distribution of death by gender, age, residence, place of death and its forensic category is presented. According to our results, the deaths due to diseases comprised 35% of all forensic autopsies. The most common cause of violent death was damage by mechanical factors (53%), followed by asphyxia (24.7%). The most common mechanical factor was vehicle crash trauma, followed by falling (22.5%). Accidents were the most common cause of death 62%, followed by suicides (31 %) and homicides 7%. Fifty-nine percent of the suicides were by hanging. The preliminary analyses of the studied data lead to several conclusions. The number of forensic autopsies was found to be progressively decreasing, but the road traffic death rate remained unchanged. Most of the forensic autopsies were performed in July. The highest death rates by age was observed in the age group between 51 and 60 years, with predominance of males over females (3.3 males: 1 females).
Maria R. Pavlova, Elina G. Dobreva, Katucha I. Ivanova, Galina D. Asseva, Ivan N. Ivanov, Peter K. Petrov, Valeri R. Velev, Ivelina I. Tomova, Maida M. Tiholova and Todor V. Kantardjiev
Introduction: Campylobacter spp. are important causative agents of gastrointestinal infections in humans. The most frequently isolated strains of this bacterial genus are Campylobacter jejuni and Campylobacter coli. To date, genetic methods for bacterial identification have not been used in Bulgaria. We optimized the multiplex PSR assay to identify Campylobacter spp. and differentiate C. jejuni from C. coli in clinical isolates. We also compared this method with the routinely used biochemical methods.
Aim: To identify Campylobacter spp. and discriminate C. coli from C. jejuni in clinical isolates using multiplex PCR assay.
Materials and methods: Between February 2014 and January 2015 we studied 93 stool samples taken from patients with diarrheal syndrome and identified 40 species of Campylobacter spp. in them. The clinical material was cultured in microaerophilic atmosphere, the isolated strains being biochemically diff erentiated (hydrolysis of sodium hippurate for C. jejuni, and hydrolysis of indoxyl acetate for C. coli). DNA was isolated from the strains using QiaAmp MiniKit (QIAGEN, Germany). Twenty strains were tested with multiplex PCR for the presence of these genes: cadF, characteristic for Campylobacter spp., hipO for C. jejuni and asp for C. coli.
Results and discussion: The biochemical tests identified 16 strains of C. jejuni, 3 strains of C. coli, and 1 strain of C. upsaliensis. After the multiplex PCR assay the capillary gel electrophoresis confirmed 16 strains of C. jejuni, 2 strains of C. coli and 2 strains of Campylobacter spp. - because of the presence of the gene cadF. C. jejuni has the gene hipO, and it is possible that this gene may not be expressed in the biochemical differentiation yielding a negative reaction as a result. In comparison, we can conclude that the genetic differentiation is a more accurate method than the biochemical tests.
Conclusion: The multiplex PCR assay is a fast, accurate method for identifi cation of Campylobacter spp. which makes it quite necessary in the clinical diagnostic practice.