The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A

Protein engineering provides an opportunity to generate new immunogens with desired features. Previously, we have demonstrated that hamster polyomavirus major capsid protein VP1-derived virus-like particles (VLPs) are highly immunogenic and can be employed for the insertion of foreign epitopes at certain surface-exposed positions. In the current study, we have designed pseudotype VLPs consisting of an intact VP1 protein and VP2 protein fused with the target antigen—cellular marker p16INK4A—at its N terminus. Both proteins coexpressed in yeast were self-assembled to pseudotype VLPs harbouring the inserted antigen on the surface. The pseudotype VLPs were used for generation of antibodies against p16INK4A that represents a potential biomarker for cells transformed by high-risk human papillomavirus (HPV). The pseudotype VLPs induced in immunized mice a strong immune response against the target antigen. The antisera raised against pseudotype VLPs showed specific immunostaining of p16INK4A protein in malignant cervical tissue. Spleen cells of the immunized mice were used to generate monoclonal antibodies against p16INK4A protein. The specificity of antibodies was proven by the immunostaining of HPV-transformed cells. In conclusion, the current study demonstrates the potential of pseudotype VLPs with inserted target antigen as a new type of immunogens to generate antibodies of high diagnostic value

Women’s Feelings about Childlessness in Two Pro-Natalist Countries

This article focuses on women’s feelings in regard to being childless/childfree in two pro-natalist countries- Lithuania and Poland. The article is based on semi-structured interviews conducted in Lithuania (N = 40) and Poland (N = 31) in 2017–2018. Comparing the experiences of two generations of women, the article reveals how social pressure functions as a mechanism of social exclusion in both countries. The analysis of circumstantial childlessness showed two ways in which women constructed their narratives: feeling normal (which prevailed in the older generation), and having ambivalent or negative reactions (which was prevalent in the narratives of both generations). The most intense emotions were revealed by women who had not had children for physiological reasons, while those women who had chosen not to have children faced strong pressure from the social environment

Barriers of Information and Communication Technologies (ICT): Narratives of Older Users and Their Facilitators

As world populations are ageing, more attention is given to the social and digital inclusion of older people. The level of ICT non-use among older adults in Lithuania is significantly higher than in Western European countries. In 2023, 29% of the Lithuanian population aged 65–74 had never used the Internet, while in most Western countries, this share varied between 2 and 8%. Insufficient research efforts in Eastern and Central Europe prompted this study to explore the barriers to ICT use in Lithuania based on qualitative semi-structured interviews with older users (N=36) and their facilitators (N=9). The difficulties revealed by the study are divided into three groups. First, the participants found it difficult to adjust to the technologies as a unique system (to understand its specific principles of functioning, to deal with information on English and technical language). Second, their psychological relationship with the unknown and complex subject have caused some problems, as they faced a spectrum of fears, insecurity, the shame of not knowing, the experience of one’s limitations, lack of patience. Third, changes that informants relate to old age like diminishing motivation to engage with innovations, learning difficulties, and bodily changes (sensitivity of fingers, weakening sight and memory) constituted additional barriers for older ICT users. To overcome the barriers, access to mentoring and consulting would be beneficial

Surveillance of cfDNA Hot Spot Mutations in NSCLC Patients during Disease Progression

Non-small cell cancer (NSCLC) has been identified with a great variation of mutations that can be surveyed during disease progression. The aim of the study was to identify and monitor lung cancer-specific mutations incidence in cell-free DNA as well as overall plasma cell-free DNA load by means of targeted next-generation sequencing. Sequencing libraries were prepared from cell-free DNA (cfDNA) isolated from 72 plasma samples of 41 patients using the Oncomine Lung cfDNA panel covering hot spot regions of 11 genes. Sequencing was performed with the Ion Torrent™ Ion S5™ system. Four genes were detected with highest mutation incidence: KRAS (43.9% of all cases), followed by ALK (36.6%), TP53 (31.7%), and PIK3CA (29.3%). Seven patients had co-occurring KRAS + TP53 (6/41, 14.6%) or KRAS + PIK3CA (7/41, 17.1%) mutations. Moreover, the mutational status of TP53 as well an overall cell-free DNA load were confirmed to be predictors of poor progression-free survival (HR = 2.5 [0.8–7.7]; p = 0.029 and HR = 2.3 [0.9–5.5]; p = 0.029, respectively) in NSCLC patients. In addition, TP53 mutation status significantly predicts shorter overall survival (HR = 3.4 [1.2–9.7]; p TP53 mutation incidence as well as a cell-free DNA load can be used as biomarkers for NSCLC monitoring and can help to detect the disease progression prior to radiological confirmation of the status