Possibilities of cytological diagnostics in gynecology. Human papillomavirus as an interdisciplinary problem

The article presents an analysis of current information on the etiology, pathogenesis, laboratory diagnosis of human papillomavirus. It is shown that at the current stage of development of laboratory diagnostics there is a reliable screening test — cytological examination of smears taken from the ecto- and endocervix. To diagnose HPV, a combination of microscopic (cytological studies) and molecular genetic (PCR) diagnostic methods is used.

Liquid technology in the cytological diagnosis of bladder pathology

Objective. To study the effectiveness of liquid cytological diagnosis of bladder cancer and its local relapses using the Cellprep Plus technology as an example.Materials and methods. We analyzed outpatient records of patients with urothelial pathology (n = 806) who underwent a urine cytology exam by the methods of liquid (n = 383) and conventional (n = 423) cytology.Results. The diagnostic sensitivity and specificity of the cytological examination method for diagnosing urothelial carcinoma using the method of liquid cytology have been found to be 93.4 % and 95.4 % respectively, which significantly exceeds the similar indices in the use of the method of conventional cytology – 42.4 % and 93.6 % respectively. The use of the method of liquid cytology considerably increases the accuracy of the cytological examination of bladder pathology and allows obtaining conclusions that coincide with the histological conclusion in 94.0 % of cases. In the use of the method of conventional cytology, the coincidence with histological findings is only 44.6 % (χ2 = 25.08, p < 0.001).Conclusion. The Cellprep Plus liquid technology standardizes the pre-analytical stage and increases the efficiency of the cytological method in the primary diagnosis and monitoring of patients with urothelial pathology. A promising direction of using the method of liquid cytology in the diagnosis of urothelial carcinoma is the development and implementation of the cytological criteria of differential diagnosis between reactive cell atypia and atypia characteristic of a malignant tumor.

Colposcopic features of cervical oncotropic papilloma-virus infection in patients with various pathology of uterine cervix

In order to estimate the colposcopic manifestations of cervical oncotropic human papilloma-virus (HPV)-infection, a total of 693 patients were examimed. Among them, there were 298 patients with benign tumors pathology, 57 patients with 1-3 Grade cervical dysplasia of mucosa, 50 patients with uterine cervix cancer, 288 healthy women. All patients underwent bimanual examination, taking of cervical smears for cytological examination and uterine cervix colposcopy. Diagnosis for HPV16/18 infection was made by the method of polimerase chain reaction. А large variety in colposcopic manifestations of HPV-infection was found, namely: areas of atypical vessels, leukoplakia sites,fields of atypical epithelium, iodine-negative sites. It was related to the influence of oncogenic types of HPV infection. In these patients,fields of atypical epithelium, atypical vessels, iodinenegative areas were observed 1.2, 2.5, 10.5 times more frequently, respectively. It was found that all varieties of papillomas occurred among patients with pathology, whereas flat condylomas presenting the most difficulties of or diagnosis prevailed in patients with cervical neoplasms and uterine cervix cancer The most pronounced colposcopic evidences of uterine cervix epithelium malignancy were observed in patients with virus-positive uterine cervix cancer.

The Potential of Optimized Liposomes in Enhancement of Cytotoxicity and Apoptosis of Encapsulated Egyptian Propolis on Hep-2 Cell Line

Purpose: Development of pharmaceutical dosage forms of natural products has gained great interest recently. Propolis is a natural product with various active compounds and multiple pharmacological activities. Its resinous nature and low bioavailability were obstacles in the optimum use of this magnificent natural product. Aim: This study evaluates the effect of using liposomes as a drug delivery system on the enhancement of the cytotoxic effect of propolis on squamous cell carcinoma cell lines (Hep-2) of head and neck. Methods: An optimized liposomal formulation of propolis was prepared using the conventional thin film hydration method 1, 2. The prepared (Hep-2) cell line was treated with different concentrations of propolis and optimized propolis liposomes for 24 h. The effect of both propolis and propolis liposomes on cell line was investigated using MTT assay, cytological examination, and nuclear morphometric analysis. The effect of the drugs on the cell apoptosis was evaluated using Annexin V. Results: The findings revealed that both propolis and propolis liposomes have a cytotoxic effect on Hep-2 cell line through induction of apoptosis. The effect was dose dependent. However, a statistically significant enhancement in propolis-mediated apoptosis on Hep-2 cells was elucidated due to encapsulation within the prepared liposomes. Conclusion: Liposome is a powerful tool for enhancing the cytotoxicity of propolis against Hep-2 cell line.

Molecular testing on serous effusion: An update

Cytological examination of the effusion fluid provides valuable information regarding the presence of malignancy. At times, it is challenging to diagnose malignant cells in serous effusion. The various ancillary techniques are available to solve the problem including immunocytochemistry, DNA ploidy, and multicolored flow cytometry. At present, the molecular tests on the effusion sample are of growing interest. The effusion sample is rich in cells and cell-free fluid that contains free DNA, cytokines, and extracellular vesicles. Molecular tests in effusion sample not only provide a diagnosis of malignancy but can also give valuable information that may be essential for the individualized therapy, management, and prognostic assessment. In this paper, we reviewed the application of the different molecular tests in the effusion sample.

The role of molecular biology in the diff erential diagnosis of pancreatic cystic neoplasias

Summary: Pancreatic cysts have been detected ever more frequently in recent years due to the advanced and wider use of imaging methods. We find them on CT or MR also in asymptomatic patients who do not have a history of any pancreatic disease. Pancreatic cystic lesions represent a wide range of pathological changes from simple cysts through precancerous lesions to malignant cysts. Accurate dia­gnosis remains difficult despite the combination of clinical status evaluation, imaging findings, and bio­chemical and cytological examination. Molecular bio­logical examination of cyst aspirate obtained by endosonographic examination increases the detection rate of mucinous cysts (KRAS/GNAS/VHL) and cysts with a high risk of malignancy (KRAS/GNAS/p53/PIK3CA/PTEN/CDKN2A/SMAD4) and optimizes therapeutic approach. Larger prospective validation studies are necessary to make this costly and limited method a routine part of clinical practice. Key words: molecular bio­logy – neoplasia – pancreatic cysts

Unusual congenital polydactyly in mini-pigs from the breeding group of the Institute of Cytology and Genetics (Novosibirsk, Russia)

The article describes a new phenomenon in the breeding group of mini-pigs at the Institute of Cytology and Genetics (ICG, Novosibirsk): polydactyly (extra digits), which is unusual because the additional digits are situated at the lateral surface of legs or at the lateral and medial ones. This anomaly was first found here in 2017 in adult animals intended for culling due to incorrect positioning of the legs caused by flexor tendon laxity and resulting in weight-bearing on the palmar surface of the proximal phalanges (“bear’s paw”). Therefore, the polydactyly of mini-pigs has a pronounced negative selection effect. A visual survey of the livestock was conducted, and a description of the detected anomaly was compiled. The polydactyly in mini-pigs is a stand-alone trait and is not part of any syndromes. Individuals with polydactyly may have extra digits either on pectoral or on pectoral and pelvic limbs. On thoracic limbs, there may be either one lateral digit or a lateral digit and a medially located rudimentary hooflet. On pelvic limbs, only lateral extra digits can occur. Anatomical and morphological analyses showed that the lateral extra digit is an anatomically complete (“mature”) structure, whereas the medial rudimentary digit consists of only a hooflet without other structures characteristic of normal digits. Cytological examination revealed no specific karyotypic features, except for Robertsonian translocation Rb 16;17 previously reported for the mini-pigs of the same livestock. Cytological findings indicated that the polydactyly and Robertsonian translocation are not linked genetically. Genealogical analysis and results of crosses are consistent with a working hypothesis of recessive inheritance of the trait. Overall, the study shows that this type of polydactyly is anatomically and morphologically unique and not typical of Sus scrofa. In this species, only polydactyly types with medial accessory toes have been described and are usually inherited as a dominant trait with incomplete penetrance. In our case, the results of test crosses indicate recessive inheritance of the trait with varying expression and incomplete penetrance, because of which poorly expressed phenotypes are not visually detectable.

Weakly supervised learning for classification of lung cytological images using attention-based multiple instance learning

In cytological examination, suspicious cells are evaluated regarding malignancy and cancer type. To assist this, we previously proposed an automated method based on supervised learning that classifies cells in lung cytological images as benign or malignant. However, it is often difficult to label all cells. In this study, we developed a weakly supervised method for the classification of benign and malignant lung cells in cytological images using attention-based deep multiple instance learning (AD MIL). Images of lung cytological specimens were divided into small patch images and stored in bags. Each bag was then labeled as benign or malignant, and classification was conducted using AD MIL. The distribution of attention weights was also calculated as a color map to confirm the presence of malignant cells in the image. AD MIL using the AlexNet-like convolutional neural network model showed the best classification performance, with an accuracy of 0.916, which was better than that of supervised learning. In addition, an attention map of the entire image based on the attention weight allowed AD MIL to focus on most malignant cells. Our weakly supervised method automatically classifies cytological images with acceptable accuracy based on supervised learning without complex annotations.