Single-objective high-resolution confocal light sheet fluorescence microscopy for standard biological sample geometries

Three-dimensional fluorescence-based imaging of living cells and organisms requires the sample to be exposed to substantial excitation illumination energy, typically causing phototoxicity and photobleaching. Light sheet fluorescence microscopy dramatically reduces phototoxicity, yet most implementations are limited to objective lenses with low numerical aperture and particular sample geometries that are built for specific biological systems. To overcome these limitations, we developed a single-objective light sheet fluorescence system for biological imaging based on axial plane optical microscopy and digital confocal slit detection, using either Bessel or Gaussian beam shapes. Compared to spinning disk confocal microscopy, this system displays similar optical resolution, but a significantly reduced photobleaching at the same signal level. This single-objective light sheet technique is built as an add-on module for standard research microscopes and the technique is compatible with high-numerical aperture oil immersion objectives and standard samples mounted on coverslips. We demonstrate the performance of this technique by imaging three-dimensional dynamic processes, including bacterial biofilm dispersal, the response of biofilms to osmotic shocks, and macrophage phagocytosis of bacterial cells

Precancerous lesions of the cervix — aetiology, classification, diagnosis, prevention

The present review introduces the aetiology and classification of cervical precancers. The principles of diagnosis based on colposcopy are reviewed. The indications for colposcopy and targeted biopsy are steps in the diagnostic process of cervical precancers. Prophylaxis of these diseases prevents cervical cancer as high-grade precancerous lesions represent a direct precursor to cervical cancer. The basics of primary and secondary prevention, the types of screening, and the behaviour of the already-alerted patients after different screenings are presented

Pregnancy and malignant diseases — principles of management

Pregnancy-associated malignant diseases introduce multiple dilemmas to the multidisciplinary boards, related to both the oncological treatment as well as to obstetrical management. The most frequent oncological diseases diagnosed during pregnancy are breast cancer, oncohematological conditions, uterine cervix cancer and skin cancers. There are different clinical scenarios: interruption of the pregnancy and further use of the most appropriate oncological strategy; it is also possible to postpone the oncological treatment for the postpartum period with a watch-and-wait strategy until the foetus is mature and the delivery is planned. The third scenario includes concurrent treatment of both conditions: use of chemotherapy, radiotherapy and surgery during an ongoing pregnancy. Choosing among these scenarios is considering many factors, including type and stage of the malignant tumour, pregnancy term, desire and informed decision of the pregnant woman to keep or interrupt the pregnancy. The current review is focused on the basic principles of the oncological modalities (surgery, chemotherapy and radiotherapy) during pregnancy as well as their influence over the pregnant woman and the foetus, over the obstetrical management and the timing and mode of delivery, delivery anaesthesia, lactation and breastfeeding from the point of view of the evidence-based medicine

Endometriosis and risk of ovarian cancer

Endometriosis is common in premenopausal women and affects about 10% of women of reproductive age. It is a benign condition but demonstrates malignant behaviour with recurrences and metastases. Its tendency to increase the risk of specific subtypes of ovarian cancer is being discussed, because they exhibit specific clinical features that distinguish them from classical ovarian cancer. Malignant transformation of endometriosis goes through its transition to atypical endometriosis. Although endometriosis-associated ovarian carcinomas have a good prognosis, adequate follow-up and monitoring after treatment of endometriosis are recommended

Heavy metals in bones from Harbour Porpoises Phocoena phocoena from the Western Black Sea Coast

During the last few years, the Western Black Sea coast has documented increase in the number of stranded marine mammals, particularly the harbour porpoise (Phocoena phocoena). This species is a subject to threats such as exposure to contaminants, fishery by-catch and introduced new marine species. The aim of this study was to analyse spatial and age trends in bone metal concentration in harbour porpoises from the Western Black Sea Coast. Selected heavy metals (Cu, Pb, Zn, Cd and Ni) were measured in bones of 33 harbour porpoises stranded along the Bulgarian Black Sea Coast from 2017. Spatially, we found higher metal levels in the harbour porpoises stranded in the Northern region compared with those from Southern region. The effect of aging was evident only for Zn content – the levels were higher in juveniles than in adults. The obtained results suggested that heavy metal contamination represent an important threat encountered by harbour porpoises

Cetacean mortality along the Bulgarian Black Sea Coast during 2017

The present study provides information on the stranding of cetaceans along the Bulgarian Black Sea Coast during 2017. Observations were made trough covering transects along the Bulgarian Black Sea Coast. A total of 198 cetaceans of three species were found - Harbor Porpoise (Phocoena phocoena), Bottlenose Dolphin (Tursiops truncatus), and Common Dolphin (Delphinus delphis). The most numerous were the harbor porpoise - 48% followed by the bottlenose dolphin - 15% and common dolphin - 8%. The most significant number of strandings was observed during July and August (54% of all cases). Cetacean strandings events were relatively highly concentrated in the area of Shabla, Varna and Pomorie

Endometrial carcinoma in patients under 40 years of age : insights from the Bulgarian cancer registry

Objectives: We aimed to investigate the overall survival (OS) of young women with endometrial cancer (EC) in Bulgaria and the impact of histological type on survival.Material and methods: This is a population-wide retrospective study of patients with EC (≤ 40 years at diagnosis) registered at Bulgarian National Cancer Registry (BNCR) between 1993 and 2020. Patients were re-classified according to the 8th edition of the TNM classification.Results: In total, 30 597 patients were registered and histologically confirmed with malignant tumors of the uterine body. From that, 29 065 of them (95%) had ECs, and the rest had sarcomas. Around 1.64% of all malignant tumors of the uterine body are diagnosed in women under the age of 40. Most of them are diagnosed in the early stage. There was no significant difference in median OS for patients diagnosed before or after 2003. In recent years there was a slight improvement in survival and patients from the last cohort of this study had a 5-year survival rate of 92.5%. Patients with favorable pathology (T1, G1/2) had no lymph node involvement at the time of diagnosis and their 10-year survival rate was 94%.Conclusions: EC in young women is a rare disease. In most cases, patients are diagnosed in early stageT1, G1/2, N0 and their prognosis is excellent. However, the lack of improvement of OS of young patients with EC in the last three decades shows the need for treatment optimization.peer-reviewe

An overview of the role of long non-coding RNAs in human choriocarcinoma

Funding: The publication of this manuscript was funded by the European Cooperation in Science & Technology Program (EU COST)–COST Action CA18117: GYNOCARE- “European network for Gynaecological Rare Cancer research: From Concept to Cure”. R.D.F., S.S., A.F., S.A.O., J.J.O., M.P.W., J.B., M.S., P.O., A.Y., M.V.-S., B.E., A.G., and J.C.-A. are Action management committee members and/or form part of Working Groups 1–3, and J.C.-A. is the Chairperson of this Action.Choriocarcinoma (CC), a subtype of trophoblastic disease, is a rare and highly aggressive neoplasm. There are two main CC subtypes: gestational and non-gestational, (so called when it develops as a component of a germ cell tumor or is related to a somatic mutation of a poorly differentiated carcinoma), each with very diverse biological activity. A therapeutic approach is highly effective in patients with early-stage CC. The advanced stage of the disease also has a good prognosis with around 95% of patients cured following chemotherapy. However, advancements in diagnosis and treatment are always needed to improve outcomes for patients with CC. Long non-coding (lnc) RNAs are non-coding transcripts that are longer than 200 nucleotides. LncRNAs can act as oncogenes or tumor suppressor genes. Deregulation of their expression has a key role in tumor development, angiogenesis, differentiation, migration, apoptosis, and proliferation. Furthermore, detection of cancer-associated lncRNAs in body fluids, such as blood, saliva, and urine of cancer patients, is emerging as a novel method for cancer diagnosis. Although there is evidence for the potential role of lncRNAs in a number of cancers of the female genital tract, their role in CC is poorly understood. This review summarizes the current knowledge of lncRNAs in gestational CC and how this may be applied to future therapeutic strategies in the treatment of this rare cancer.publishersversionpublishe

Endometrial carcinoma in patients under 40 years of age: insights from the bulgarian cancer registry

Objectives: We aimed to investigate the overall survival (OS) of young women with endometrial cancer (EC) in Bulgaria and the impact of histological type on survival. Material and methods: This is a population-wide retrospective study of patients with EC (≤ 40 years at diagnosis) registered at Bulgarian National Cancer Registry (BNCR) between 1993 and 2020. Patients were re-classified according to the 8th edition of the TNM classification. Results: In total, 30 597 patients were registered and histologically confirmed with malignant tumors of the uterine body. From that, 29 065 of them (95%) had ECs, and the rest had sarcomas. Around 1.64% of all malignant tumors of the uterine body are diagnosed in women under the age of 40. Most of them are diagnosed in the early stage. There was no significant difference in median OS for patients diagnosed before or after 2003. In recent years there was a slight improvement in survival and patients from the last cohort of this study had a 5-year survival rate of 92.5%. Patients with favorable pathology (T1, G1/2) had no lymph node involvement at the time of diagnosis and their 10-year survival rate was 94%. Conclusions: EC in young women is a rare disease. In most cases, patients are diagnosed in early stageT1, G1/2, N0 and their prognosis is excellent. However, the lack of improvement of OS of young patients with EC in the last three decades shows the need for treatment optimization

Adenosquamous carcinoma of the uterine cervix – impact of histology on clinical management

Introduction: Historically, the incidence rate of cervical cancer (CC) in Eastern Europe and particularly in Bulgaria has constantly been higher than that in the other European countries. Adenosquamous carcinoma (ASC) is a rare histological subtype of CC with incidence rate of less than 6 per 100,000. We aimed to analyze the epidemiology and prognosis of all Bulgarian patients with ASC, registered at the Bulgarian National Cancer Registry (BNCR), and to compare patients’ characteristics and outcomes with those of patients, treated at a large specialized institution – the Department of Gynecologic Oncology, University Hospital in Pleven, Bulgaria. Materials and Methods: This is a retrospective study of all cases of ASC, registered at the BNCR for a 10-year period of time. The Kaplan–Meier analysis with Log rank test was used to estimate the significant differences. Results: The incidence rate of ASC was calculated as 3.2% of all CC registered in BNCR and 4.97% of all stage I patients, treated in our department. The 5-year overall survival (OS) rate of all patients with ASC tumors from the registry was 50.5%. A total of 171 (48.4%) of the patients had T1 tumors and a 5-year OS of 67.1%. Lymph node status was a significant prognostic factor for OS (p=0.001). Thirty-one patients with T1 tumors and ASC histology were treated in our department for the same period of time. Lymph node metastases were found in 10 of them (32.2%). The 5-year observed OS in ASC group was 74.19%. Conclusion: The histological subtype of cancer of the uterine cervix has an impact on prognosis and should not be simply considered as a descriptive characteristic but a poor prognostic feature and should be an integral part of the decision-making in clinical management of patients.peer-reviewe