Thrombotic disorders are some of the main comorbidities in cancer patients. So far, research has indicated that thrombin, a key regulator of hemostasis, contributes to cancer progression. However, data on its origin in tumor microenvironments remain elusive. Based on previous research, we analyzed the RNA and protein expression of prothrombin, a precursor of thrombin, in selected colorectal cancer (CRC) cell lines. Since the effect of prothrombin in cancer development has not been previously reported, we treated the cells for 24 h and 48 h with different prothrombin concentrations and assessed the effect on cell proliferation and migration. Our results show that the tested CRC cell lines expressed prothrombin and that prothrombin inhibited proliferation and migration. The presented results suggest that prothrombin may contribute to CRC etiopathology and could serve as a potential diagnostic biomarker and therapeutic target. The mechanisms underlying prothrombin expression in cancer cells, potential prothrombin activation, and the underlying processes driving the described effects warrant further investigation.

Setbacks and challenges are an unavoidable part of everyday life. As a clinical and research community, we can be proud of how we rallied to counter the recent COVID-19 pandemic. However, we are now faced with war, natural disasters, and global inflationary pressures that are likely to have a sustained impact on the lives of researchers and clinicians, particularly our early career (EC) professionals [1]. All too familiar issues such as increased workload and childcare responsibilities and decreased funding, mentorship, and networking opportunities continue to challenge our EC community [2,3].

INTRODUCTION:Prothrombin Belgrade mutation is the result of the c.1787G>A substitution in the prothrombin gene. It is located in the antithrombin and sodium binding site and leads to impaired inactivation of thrombin by antithrombin, resulting in antithrombin resistance and thrombotic disorders. However, it negatively affects sodium binding and may have hypocoagulant effects. Considering that prothrombin Belgrade mutation mechanism is still not fully elucidated and that sodium binding is important for thrombin affinity towards fibrinogen, our aim was to determine whether this mutation affects fibrin clot formation and lysis.METHODS:Using HEK293T cell line, recombinant wild type and mutated prothrombin were generated by transient transfection. Samples that correspond to plasma of a non-carrier, heterozygous and homozygous carriers were reconstituted using prothrombin deficient plasma and recombinant proteins. Reconstituted samples were used in OHP assay (Overall Hemostasis Potential) to determine kinetic profiles of coagulation and fibrinolysis. Clot turbidity assay was performed to observe kinetics of clot formation and lysis more closely. Fibrin clots formed in reconstituted plasma samples were analyzed by confocal microscopy to determine density of fibrin network. Fibrin clots were additionally observed using electron microscopy to determine thickness of individual fibrin fibers.RESULTS:No significant difference found in OHP, OCP, OFP, and fibrin network density between wild type, heterozygous, and homozygous carrier reconstituted plasma samples. There were significant differences between samples for slope and slope time parameters in kinetic profiles and fibrin fiber thickness.CONCLUSIONS:Results indicate that prothrombin Belgrade mutation has no significant impact on fibrinolysis, however it may affect kinetics of clot formation and its architecture.

Antithrombin resistance is a rare subtype of hereditary thrombophilia caused by prothrombin gene variants, leading to thrombotic disorders. Recently, the Prothrombin Belgrade variant has been reported as a specific variant that leads to antithrombin resistance in two Serbian families with thrombosis. However, due to clinical data scarcity and the inapplicability of traditional genome-wide association studies (GWAS), a broader perspective on molecular and phenotypic mechanisms associated with the Prothrombin Belgrade variant is yet to be uncovered. Here, we propose an integrative framework to address the lack of genomic samples and support the genomic signal from the full genome sequences of five heterozygous subjects by integrating it with subjects' phenotypes and the genes' molecular interactions. Our goal is to identify candidate thrombophilia-related genes for which our subjects possess germline variants by focusing on the resulting gene clusters of our integrative framework. We applied a Non-negative Matrix Tri-Factorization-based method to simultaneously integrate different data sources, taking into account the observed phenotypes. In other words, our data-integration framework reveals gene clusters involved with this rare disease by fusing different datasets. Our results are in concordance with the current literature about antithrombin resistance. We also found candidate disease-related genes that need to be further investigated. CD320, RTEL1, UCP2, APOA5 and PROZ participate in healthy-specific or disease-specific subnetworks involving thrombophilia-annotated genes and are related to general thrombophilia mechanisms according to the literature. Moreover, the ADRA2A and TBXA2R subnetworks analysis suggested that their variants may have a protective effect due to their connection with decreased platelet activation. The results show that our method can give insights into antithrombin resistance even if a small amount of genetic data is available. Our framework is also customizable, meaning that it applies to any other rare disease.

Bronchial epithelial cells are exposed to environmental influences, microbiota, and pathogens and also serve as a powerful effector that initiate and propagate inflammation by the release of pro-inflammatory mediators. Recent studies suggested that lung microbiota differ between inflammatory lung diseases and healthy lungs implicating their contribution in the modulation of lung immunity. Lactic acid bacteria (LAB) are natural inhabitants of healthy human lungs and also possess immunomodulatory effects, but so far, there are no studies investigating their anti-inflammatory potential in respiratory cells. In this study, we investigated immunomodulatory features of 21 natural LAB strains in lipopolysaccharide (LPS)-stimulated human bronchial epithelial cells (BEAS-2B). Our results show that several LAB strains reduced the expression of pro-inflammatory cytokine and chemokine genes. We also demonstrated that two LAB strains, Lactobacillus brevis BGZLS10-17 and Lb. plantarum BGPKM22, effectively attenuated LPS-induced nuclear factor-κB (NF-κB) nuclear translocation. Moreover, BGZLS10-17 and BGPKM22 reduced the activation of p38, extracellular signal-related kinase (ERK), and c-Jun amino-terminal kinase (JNK) signaling cascade resulting in a reduction of pro-inflammatory mediator expressions in BEAS-2B cells. Collectively, the LAB strains BGZLS10-17 and BGPKM22 exhibited anti-inflammatory effects in BEAS-2B cells and could be employed to balance immune response in lungs and replenish diminished lung microbiota in chronic lung diseases.

Early career professionals in the field of thrombosis and hemostasis may experience significant gender‐dependent challenges across research, clinical practice, industry, and beyond. Individuals who identify as women, hereafter referred to as women, may face systemic biases and discrimination that hinder career advancement and leadership opportunities. These barriers are more pronounced in marginalized groups, including racial and ethnic minorities and the LGBTQIA+ community. Here, we highlight some of the challenges disproportionally impacting early career women in our community and discuss potential strategies that might begin to close gender‐related gaps. Just as women with bleeding disorders may go undiagnosed and undertreated because of the stigma surrounding heavy menstrual bleeding,1.Denis C.V. Susen S. Lenting P.J. von Willebrand disease: what does the future hold?.Blood. 2021; 137: 2299-2306Crossref PubMed Scopus (9) Google Scholar women in science may encounter similar social and cultural discrimination. Harmful stereotypes and biases in the workplace are present at all career stages. Women often receive less credit for their management, leadership, and research contributions. As an example, lack of credited research contributions at an early career stage leads to a drastic reduction in authorship, with women being recognized on publications at almost half the rate of men.2.Ross M.B. Glennon B.M. Murciano‐Goroff R. Berkes E.G. Weinberg B.A. Lane J.I. Women are credited less in science than are men.Nature. 2022; 608: 135-145Crossref PubMed Scopus (24) Google Scholar A lack of scientific acknowledgement can start a vicious cycle whereby women scientists receive fewer citations and less public recognition. This may lead to failure to gain recognition within specific fields, to achieve promotion and tenure, and to receive funding. Therefore, it is not surprising that women researchers make up approximately half of the workforce but are less represented in senior faculty positions. Discrimination and harassment can occur at all stages of career development. In surveys of trainees in basic and clinical science, women respondents were significantly more likely to report sexual harassment and discrimination than their male counterparts.3.Woolston C. PhDs: the tortuous truth.Nature. 2019; 575: 403-406Crossref PubMed Scopus (114) Google Scholar, 4.McClain T. Kammer‐Kerwick M. Wood L. Temple J.R. Busch‐Armendariz N. Sexual harassment among medical students: prevalence, prediction, and correlated outcomes.Workplace Health Saf. 2021; 69: 257-267Crossref PubMed Scopus (10) Google Scholar Similarly, women pediatric hematology/oncology fellows were more likely to be mistaken as non‐physicians and be second‐guessed by colleagues than male fellows, which contributed to feelings of insecurity about their positions.5.Balanchivadze N. Passalacqua K.D. Askar S. et al.Self‐reported gender bias encountered by hematology and oncology fellows.J Clin Oncol. 2021; 39Crossref Google Scholar Unsurprisingly, sexual harassment is associated with depression and decreased academic engagement. Furthermore, reporting systems for bias or harassment are lacking in many parts of the world, and many women fail to report concerns due to fear of retribution. Early career professionals are at particularly high risk because of their sense of dependency on and intimidation by their mentors and senior colleagues. Achieving equal rights and opportunities for women in our community is impossible unless their fundamental human rights are guaranteed. Rights of access to lifesaving medical care, to one's bodily autonomy, and to family planning are the cornerstone of equal access to education and career opportunities. The recent decision by the United States Supreme Court to overturn access to abortion is just one example of how women's freedom and choices remain under threat. In countries with limited healthcare access, socio‐cultural stigmas, and/or strong anti‐abortionist views, early career women in science face restrictions on opportunities and access to education, widening the professional gap between themselves and their male counterparts. Those in historically marginalized and low income communities who already face many other forms of discrimination are disproportionally impacted by such barriers. Early career professionals who start families continue to encounter barriers at the institutional level. Many universities have yet to adopt clear policies regarding parental leave. Access to childcare support, remote work, or flexible working hours is not always guaranteed, even between different departments at the same institution. Men's access to paternal leave is often limited, which reinforces traditional gender biases resulting in women disproportionately bearing the responsibility for caring for young children. Early career researchers who take time away from the workplace for parental leave may not have their research contributions appropriately recognized during their absence or even after their return. Similarly, those returning from parental leave on a part‐time basis may feel obliged to work longer hours, even if unpaid, to complete their research goals. Meanwhile, women who decide not to have children are often judged for this personal decision during interviews, consideration for promotion or assignment of work‐related responsibilities; a problem rarely faced by men. Conversely, younger women who have not had children may be less competitive in the workplace because prospective employers may anticipate their future requirements for parental leave and part‐time work. These systemically imposed barriers represent a significant challenge in women's quest for equitable representation in science. Although these global challenges may seem overwhelming, there are several steps our community can take to mitigate the barriers facing early career women in the thrombosis and hemostasis fields. The ISTH, through its diversity, equity, and inclusion committee, has taken a leading role in increasing representation of early career women. Today, women make up 56% of the membership at the early career level (students, postdocs, medical residents, and associate members). The ISTH has increased the inclusion of women as speakers and moderators at educational events, on editorial boards and reviewer panels for JTH and RPTH, and as invited authors for papers in these journals (Figure 1). These efforts were recently highlighted in an editorial written by Dr. Mary Cushman, editor‐in‐chief of RPTH, and we applaud them as measures that will create a more inclusive culture and reduce bias in the peer‐review process.6.Cushman M. Are women equal? Considering impact of therapeutic abortion bans on science.Res Pract Thromb Haemost. 2022; 6Abstract Full Text Full Text PDF Scopus (1) Google Scholar Careful attention must continue to be paid to gender representation across our community, including recognizing the conscious and unconscious biases faced by women and non‐binary individuals. To increase awareness, the Early Career Committee holds educational events to help members address and overcome their biases. As the leader in the thrombosis and hemostasis field, the ISTH can advocate for clear guidelines regarding parental leave and reporting bias and harassment at institutions around the world. Practical initiatives such as access to childcare and breastfeeding‐friendly facilities at ISTH Congresses are also paramount. These initiatives will improve the educational and training landscape for women and other marginalized groups in science, thereby enriching the diversity of the ISTH and enhancing scientific and medical innovation.

Background/ Aim: Thrombin plays significant roles in various types of cancer. However, the expression levels of prothrombin, the thrombin precursor, in cancer remain unclear. Variants of the 3'end of the prothrombin gene lead to increased prothrombin expression. This study aimed to analyze prothrombin 3'end gene variants in colon tumor and adjacent normal tissue samples. Materials and Methods: The study group consisted of 93 patients suffering from colon adenocarcinoma. The 3'end of the prothrombin gene was analyzed by DNA sequencing. Results: Three variants, all previously associated with increased prothrombin expression were detected. Frequency of the FII 19911G allele was 46.77% and 47.85% in tumor and normal tissue, respectively. For the FII 20210A allele, the detected frequencies were 2.15% and 1.61%, respectively. The frequency of the FII c.1824T allele was 0.54% in both tissues. Four patients showed different genotypes in tumor and normal tissue. Conclusion: Prothrombin 3' end gene variants may play a role in colorectal cancer.

The link between thrombotic disorders and cancer has been known for over 150 years, although the precise mechanism of this relationship has not yet been resolved. Current data show that thrombin has a significant role in cancer metabolism, invasiveness, adhesion and survival. However, data regarding the expression of the thrombin precursor prothrombin in various cancer cell lines are scarce. Therefore, it was our objective to determine whether common cancer-derived cell lines (Caco-2, MCF-7, SK-BR-3, U-87 and U-251) express prothrombin. The prothrombin RNA expression level was assessed by qPCR, and the presence of prothrombin was analyzed by Western blot analysis. Our results show that Caco-2 cells originating from colorectal adenocarcinoma express prothrombin, whereas other analyzed cell lines do not. Our results provide a background for further research into the role of (pro) thrombin in cancer etiopathology.