Evaluation of cleaved caspase-3 and Ki-67 index on diagnostic biopsy in response to neoadjuvant chemotherapy in the context of post-treatment tumour ypT stage, ypN stage, grade, and molecular subtype.

Introduction: Breast carcinoma is a heterogeneous disease, characterised by diverse clinical behaviour. The aim of this study was to assess how cleaved caspase-3 and Ki-67 index, evaluated on diagnostic biopsy, are related to response to neoadjuvant chemotherapy in the context of molecular subtype, post-treatment tumour, N category, and grade. Material and methods: A retrospective analysis was carried out among 110 breast cancer patients. Ki-67 levels and caspase-3 expression on diagnostic biopsy were explored regarding their relation to tumour grade and molecular subtype, ypT, ypN categories, and T and N categories according to Sataloff tumour response evaluation. Results: A statistically significant relationship was found between Ki-67 levels and tumour grade K-W = 24.2932, p < 0.0001; molecular subtype K-W = 28.5439, p < 0.00000967538; size and invasion of the primary tumour after neoadjuvant chemotherapy K-W = 11.7944, p < 0.0377169; caspase-3 expression after neoadjuvant therapy, evaluated according to the Sataloff classification χ2 = 5.97, df = 1, p = 0.0145. Discussion: No significant difference was found between Ki-67 expression in patients with pathological complete response, compared to those with partial and no response, a statistically significant difference in cases with different molecular subtype, histology grade, and tumour stage after neoadjuvant therapy. Cleaved caspase-3-positive breast cancer cases are often better responders to neoadjuvant therapy, but with no significant correlation to molecular subtype, high-grade categories, or tumour stage. Conclusions: The caspase-3 and Ki-67 index on diagnostic biopsy are related to post-neoadjuvant treatment prognostic factors (ypT stage, grade), proving them useful for prediction of treatment response to neoadjuvant therapy and further patient management.

First case of ocular dirofilariosis in Bulgaria caused by gravid female Dirofilaria repens.

Dirofilariosis caused by Dirofilaria (Nochtiella) repens is recorded sporadically among people in Europe, Asia and Africa. Still a worldwide controversy exist upon human parasite hosting. Herein, the first case of ocular dirofilariosis in Bulgaria caused by gravid female is presented. A single nematode was removed from the eye of the 76- year-old patient in the course of cataract surgery. Microscopic examination of the histological slides revealed microfilariae in the parasite's uterus, but not in the blood. Knott's method for detection of microfilariae in the peripheral blood was negative and the rest of laboratory blood and biochemical tests were within reference limits. A comprehensive review of the etiology, pathogenesis and clinical presentation of Dirofilaria repens infection is also highlighted.

A case of a child with four parasites in Bulgaria.

Children are more susceptible to intestinal parasitic diseases than adults due to their lack of hygiene habits and close social interactions with their peers. We present a case of a 5-year-old boy with diarrhea and abdominal pain. The stool sample and perianal swab were examined in LINA laboratory, Burgas, Bulgaria. We detected four parasites Giardia (Lamblia) intestinalis, Blastocystis hominis, Hymenolepis nana and Entamoeba coli. Microbiological stool examination shows negative results for Candida spp., Salmonella spp. and Shigella spp. Patient's peripheral blood tests were normal. The calprotectin in faeces was tested establishing slightly increased values. The presence of four parasites in one patient in Bulgaria (in the 21st century) is a rare case. Lambliosis and hymenolepiosis are more likely to be found in this patient's region. Following the presence of Entamoeba coli and the transmission mechanism of the other three parasites it can be concluded that the child's hygiene is poor. This patient reveals typical clinical picture with mild symptoms and normal paraclinical results. The presence of parasites in a definitive host is not necessarily related with the development of pathological departures. The fight with parasitic diseases can be successful with a broad integral approach - recognizing and diagnosing the parasites promptly and using radical and suitable treatment.

Estimation of Ixodes ricinus (Acari: Ixodidae) Populations of Kaylaka Park in the Town of Pleven, Bulgaria.

(1) Background: Ticks are vectors of a large number of pathogenic microorganisms, which cause serious diseases in both humans and animals. Kaylaka Park is located in northern Bulgaria close to the city of Pleven. Part of the park is urbanized and visited daily by many citizens. The aim of our study was to determine the presence and distribution of hard ticks in the park area by surveying and comparing four urbanized with four wild areas. (2) Methods: Ticks were collected by flagging from 2016 to 2020 during the spring-summer season (March-July). Air temperature, relative humidity, collection time and flagging area were measured during the campaign. (3) Results: A total of 622 ticks were collected: 285 females (46%), 272 (44%) males and 64 (10%) nymphs. All were identified as Ixodes ricinus. Wild areas showed statistically significant higher values of ticks collected per minute (p = 0.009) and nymph densities (p = 0.003) compared to urbanized sampling sites. Other densities indices did not have a significant difference between urban and wild areas. Highest numbers of Ixodes ticks were collected at a temperature of 20 °C and at 60% relative humidity. The active questing began in March, peaked in end of April and declined in June. (4) Conclusions: In the present study, we found that ecological factors in the Kaylaka Park area are favourable for the development and distribution of tick populations. The results give us reason to consider that there is a high risk to visitors from tick bites in the Kaylaka Park area.

Syphilis in the Pleven region, Bulgaria: a retrospective study.

INTRODUCTION: Syphilis is a sexually transmitted infection whose significance to public health has not lost its relevance. Changes in the epidemiology of the disease have been observed in the last decade. This study describes syphilis trends in the Pleven region (Bulgaria). METHODS: Data from 2009 to 2018 were extracted from the National Center for Public Health and Analyses in Sofia and the Regional Health Inspectorate in Pleven and analyzed. RESULTS: There were 3,602 cases of syphilis during the 10-year period in Bulgaria. The incidence rate (per 100,000) during the period observed varied from 4.2 in 2011 to a maximum rate of 7.5 in 2017. For the same period, 119 cases of syphilis were reported in the Pleven region, with highest incidence rate registered in 2009 (7.2), and the lowest rate registered in 2014 (2.6). The disease was more common in men. All ages are affected, but the largest group is 25 to 29 years old. During the study period, 219 cases of congenital syphilis were registered in the country, three of them in Pleven region. CONCLUSION: Retrospective observational data confirmed a rising trend in syphilis cases, especially among younger people. Further study of risk groups and the expansion of promotional activities among adolescents is needed.

An overview of cervical cancer epidemiology and prevention in Bulgaria.

INTRODUCTION: Cervical cancer ranks as the fourth leading cause of female cancer in Bulgaria. The objective of the study was to describe the current burden of cervical cancer in Bulgaria for a one-year period and prevention options. METHODS: In this retrospective epidemiological study, online database from National Statistical Institute-Sofia, National Center of Public Health and Analyses-Sofia, National Center of Infectious and Parasitic Diseases-Sofia for one year were analyzed. Annual data were presented in numbers and per 100 000 of population. RESULTS: A total of 15,759 women in Bulgaria were diagnosed with cervical cancer in 2018. Morbidity is 23.5 of 100 000 population. Mortality of malignancy of the cervix in 2018 per 100 000 populations was 8.4 among women. Differences of mortality have been recorded as per statistical regions. A low immunization range has been recorded of vaccine against human papilloma virus (HPV) - 7.2% (girls 13 years of age) and 12.8% (girls 12 years of age) for first dose and 11.9% (girls 12 year of age) for second dose in 2019. CONCLUSIONS: High morbidity and mortality from cervical cancer were registered in Bulgaria. The immunization coverage was low. The strategy for disease prevention through mandatory screening and mandatory immunizations needs to be rethought.

Single-Center Study of Lymphoepithelioma-Like Carcinoma of Uterine Cervix over a 10-Year Period.

Background and objectives: Lymphoepithelioma-like carcinoma (LELC) is a histological type of malignant tumor arising from the uncontrolled mitosis of transformed cells originating in epithelial tissue. It is a rare subtype of squamous cell carcinoma of the uterine cervix. There are significant differences in frequency, mean age, viral status, and outcomes in Asian or Caucasian patients. Materials and Methods: A retrospective study of all cases of lymphoepithelioma-like carcinoma of the cervix at the Clinic of Oncogynecology, University Hospital, Pleven, Bulgaria between 1 January 2007 and 31 December 2016 was performed. All patients were followed-up till March 2019. We analyzed some clinical characteristics of the patients, calculated the frequency of lymphoepithelioma-like carcinoma of the cervix from all patients with stage I cervical cancer, and looked at the overall survival rate, the 5-year survival rate, and the correlation between overall survival, lymph node status, and the size of the tumor. Results: The frequency of lymphoepithelioma-like carcinoma was 3.3% for all cases with cervical carcinoma at stage I. The mean age of the patients with LELC was 49.6 years (range 32-67). Fourteen patients (82.4%) were in the FIGO IB1 stage, three patients (17.6%) were in the FIGO IB2 stage. Lymph nodes were metastatic in three patients (17.6%), non-metastatic in 13 patients (76.5%), and unknown in one patient. The overall survival rate was 76.47% for the study period and the 5-year survival rate of the patients that were followed-up until the 5th year (14 patients) was 69.23%. Conclusions: Lymphoepithelioma-like carcinoma is a rare SCC subtype, but it could be more frequent among western patients than previously thought. Our results do not confirm the data showing low risk of lymph metastasis and good prognosis of LELC, which is why we think that the treatment in these cases has to be more aggressive than is reported in the literature.

Trends in the Acute Hepatitis B and Acute Hepatitis C in Bulgaria.

INTRODUCTION: Bulgaria joined European Union (EU) on 1 January 2007. Since the accession all regulations, directives, decisions, recommendations, and opinions of the European Parliament, the Council of the European Union, and the European Commission are being implemented. AIM: The purpose of this study was to present the morbidity of acute hepatitis B virus (HBV) and acute hepatitis C virus (HCV) in Bulgaria before and after accession to the EU. MATERIALS AND METHODS: A retrospective study was performed. The morbidity of acute HBV and acute HCV infections in Bulgaria was analyzed over a period of sixteen years (2000-2016). The collected data were based on the National Center of Infectious and Parasitic Diseases (NCIPD) and the National Center of Public Health and Analyses (NCPHA). RESULTS: Between 2000 and 2016, 11038 cases of acute HBV infection and 1681 cases of acute HCV infection were reported in Bulgaria. Before the accession to EU, the morbidity rates of acute HBV and acute HCV infections were 12.77 cases per 100 000 population (95% CI: 11.45-13.97) and 1.52 cases per 100 000 population (95% CI: 1.17-1.96), respectively. After the accession to the EU, the morbidity rates of acute HBV and acute HCV infections were 5.29 cases per 100 000 population (95% CI: 3.86-6.73) and 1.14 cases per 100 000 population, respectively. CONCLUSION: Analysis of the data suggests that there is a tendency for lower morbidity rates of acute HBV. The situation with acute HCV is relatively stable over the years regardless of EU membership.

Seroprevalence of IgG Antibodies against Mumps in Bulgarian Children under 18 Years of Age.

OBJECTIVE: The objective of this study was to determine the prevalence of IgG antibodies against mumps in a representative stratified sample of children under 18 years of age in Pleven district, Bulgaria. STUDY DESIGN: Cross sectional study. MATERIALS AND METHODS: A cross-sectional, sero-epidemiologic survey of 132 children aged under 18 years was conducted in Pleven district. They were distributed into two age groups: from 0 to 11 years old (n=80) and from 12 to 18 years old (n=52). An enzyme immunoassay (EIA) was performed for indirect evidence of specific IgG antibodies in the serum. RESULTS: The results showed that 41% of the investigated children were negative in terms of mumps IgG antibodies, 11% had borderline values and the remaining, approximately 48%, were positive. In the group of children aged 0 to 11 years who had received one dose of the mumps vaccine, 50% were negative for mumps IgG antibodies. In the second group of children aged 12 to 18 years who had received two vaccine doses, the percentage of participants negative for mumps IgG antibodies decreased significantly. CONCLUSION: Our observations emphasise the need to conduct expanded sero-epidemiological investigations of the populations most affected by mumps and to immunise susceptible individuals.

Severe leptospirosis observed in a man who had just returned from abroad.

Leptospirosis, a re-emerging zoonosis caused by pathogenic Leptospira, has a low incidence in Bulgaria. This paper reports a case of leptospirosis in Pleven, Bulgaria, in which the subject was infected after wading through irrigative canal in northern Greece. Two days later, he had a fever, myalgia and vomiting followed by jaundice, darkness of urine and oliguria. The patient was admitted to Clinic of Infectious Diseases at University Hospital-Pleven after returning to Bulgaria. The history and laboratory findings suggested icterohaemorrhagic leptospirosis. Penicillin G was prescribed and intensive supportive treatment was initiated. Dialysis was performed two hours after admission and was followed by poliuric stage of acute renal failure (peak urine output 16 600 mL/day). Microaglutination test (MAT) for sero-diagnosis was positive (L. hardjo 1:1600, L. icterohaemorrhagiae 1:800). The patient was discharged after sixteen days with improved renal and liver functions. In conclusion, The probability of leptospirosis should not be ignored in patients with fever after returning from abroad. The prompt dialysis and adequate treatment improve prognosis.

Icterohaemorrhagic leptospirosis in patients with history of alcohol abuse - report of two cases.

Leptospirosis is a re-emerging zoonosis caused by spirochetes from the genus Leptospira and is typically associated with rural settings. Transmission occurs via contact with urine from infected animals; incubation period ranges from 4 days to 4 weeks. The clinical spectrum of leptospirosis may be mild and self-limited or severe with jaundice, renal failure, and bleeding manifestations (icterohaemorrhagic leptospirosis, so called Weil's disease). Mortality in severe forms remains high even when optimal treatment is provided. Early clinical suspicion and laboratory confirmation of leptospirosis is essential since delays in diagnosis may increase mortality. Alcohol-related toxicity and alcoholic hepatitis are common pathological processes, which can occasionally produce clinical syndromes similar to leptospirosis. There are few reports regarding the clinical course of leptospirosis in chronic alcoholics. Here, we describe two patients with Weil's disease, in whom alcohol abuse caused therapeutic difficulties. One of the cases was with lethal outcome.