Evaluation of T-cell clonality by anti-TRBC1 antibody-based flow cytometry and correlation with T-cell receptor sequencing.

Flow cytometry (FC) incorporating the T-cell receptor ß constant chain-1 (TRBC1) has been recently proposed as a new standard in T-cell clonality assessment. While early studies demonstrated high sensitivity in samples with conspicuous tumour burden, performance in real-world samples, including those with low tumour burden and correlation with molecular methods has been limited. We evaluated TRBC1-FC performance and correlated the results with high-throughput TRB sequencing and a targeted next-generation sequencing gene panel. Our cohort consisted of 90 evaluable samples from 57 patients. TRBC1-FC confirmed T-cell clonality in 37 out of 38 samples (97%) that were involved in a mature T-cell neoplasm (MTCN). T-cell clonality was also identified in nine samples from patients lacking a current or prior diagnosis of MTCN, consistent with the emerging entity T-cell clonality of uncertain significance. TRBC-FC was polyclonal in all samples and negative for disease involvement by standard pathology assessment. However, correlation with TRB sequencing in 17 of these samples identified two cases that harboured the known clonal sequence from index testing, indicating the presence of measurable residual disease not otherwise detected. Our study provides real-world correlative validation of TRBC1-FC, highlighting the strengths and limitations pertinent to its increasing implementation by general diagnostic laboratories.

"You feel like you're fairly disadvantaged with an advert over your head saying 'in final years of reproduction'": social egg freezing, dating and the (unequal) politics of reproductive ageing.

Recent decades have seen an increasing gap occurring between the 'desired' and 'actual' family size of middle-class and professional women. This situation of 'unrealised fertility' and 'incomplete families' carries implications at a population, but also couple and individual level. This paper explores how middle-class professional women make decisions about partnering and parenthood and how these are shaped by a contemporary neoliberal feminist discourse which articulates the possibility of 'having it all' through engagement in careful life planning, appropriate self-investment, and by drawing on new technologies of reproductive biomedicine. Informed by semi-structured interviews with UK and US women conducted at two different points in time, it explores how they approach and experience the process of relationship formation in the face of age-related fertility decline. It also examines how the use of social egg freezing shape their romantic and family building expectations but also their interactions with (potential) partners. In doing so, it explores how gendered cultural dating scripts and unequal gender power relations shape the formation and progression of intimate relationships in a manner which can disempower women as they age. It therefore questions whether egg freezing may be the 'great equaliser' that some have hoped.

Computational flow cytometry provides accurate assessment of measurable residual disease in chronic lymphocytic leukaemia.

Undetectable measurable residual disease (MRD) is associated with favourable clinical outcomes in chronic lymphocytic leukaemia (CLL). While assessment is commonly performed using multiparameter flow cytometry (MFC), this approach is associated with limitations including user bias and expertise that may not be widely available. Implementation of unsupervised clustering algorithms in the laboratory can address these limitations and have not been previously reported in a systematic quantitative manner. We developed a computational pipeline to assess CLL MRD using FlowSOM. In the training step, a self-organising map was generated with nodes representing the full breadth of normal immature and mature B cells along with disease immunophenotypes. This map was used to detect MRD in multiple validation cohorts containing a total of 456 samples. This included an evaluation of atypical CLL cases and samples collected from two different laboratories. Computational MRD showed high correlation with expert analysis (Pearson's r > 0.99 for typical CLL). Binary classification of typical CLL samples as either MRD positive or negative demonstrated high concordance (>98%). Interestingly, computational MRD detected disease in a small number of atypical CLL cases in which MRD was not detected by expert analysis. These results demonstrate the feasibility and value of automated MFC analysis in a diagnostic laboratory.

Conceptualising women's motivations for social egg freezing and experience of reproductive delay.

As the average age of motherhood in many Western countries continues to rise, the spectacle of the older mother and the trend towards delayed childbearing has been the subject of much public debate and interest. Concurrent to this trend has been the development and use of a new form of fertility preservation - social egg freezing - a technology which by its very nature is meant to enable reproductive delay. Whilst previous studies have been able to provide insights into the complex and often interrelating structural, economic, and relational factors shaping the timing of motherhood, and in some cases women's use of social egg freezing, fewer studies have clearly demonstrated the way these factors themselves, as well as the accounts of individual women, can be seen as being shaped by ideological and discursive forces. Drawing on interviews with 31 users of social egg freezing this article will demonstrate how women's accounts of reproductive delay and use of egg freezing technology can be seen as being shaped by neoliberal rationality, heteronormativity, discourses of 'appropriate parenting' and gendered ideologies of parenthood.

Oocyte cryopreservation for social reasons: demographic profile and disposal intentions of UK users.

A small number of studies from the USA and Europe have provided some data on the profile and characteristics of women who have undergone oocyte cryopreservation for what has been termed elective, social or non-medical reasons; however, little is known in a UK context about which women are undergoing oocyte cryopreservation or their reproductive intentions and actions after the procedure. Drawing on data from an exploratory study of 23 UK resident women who had undergone social oocyte cryopreservation, the demographic profile of these women, their reproductive intentions and actions are discussed, as well as their attitudes and intentions towards their cryopreserved oocytes should they never require them in treatment. The study found that, at the time of oocyte cryopreservation, women were on average 36.7 years of age, were university educated, with 65% of the sample holding further postgraduate or professional qualifications. Fifty-seven per cent of the participants were in professional employment. All participants identified as heterosexual and 87% were not in a relationship at the time of cryopreserving their oocytes. Most (88%) participants stated that they would donate unwanted oocytes to research or to other women for use in fertility treatment should they never require them.

Women's experience of social egg freezing: perceptions of success, risks, and 'going it alone'.

Very few studies have explored women's experiences of social egg freezing and the limited primary research on this topic has suggested that users find the process of freezing eggs emotionally challenging. We, therefore, undertook semi-structured interviews with 31 women who identified as undergoing egg freezing for social reasons in order to explore how female users of social egg freezing technology reported their experience of freezing eggs for 'social' reasons. Interviews lasted between 40 minutes and 2 hours, were audio recorded and transcribed verbatim. Data were analyzed using thematic analysis assisted by Nvivo 10. Women employed multiple concepts of egg freezing 'success'. They reported a lack of detailed discussion of post-freezing processes and outcomes in their encounters with clinicians, and, contrary to the recommendations of professional associations, were not given clinic or age-specific information. Few women perceived freezing as involving physical risks. However, many participants reported the process of egg freezing as emotionally challenging, primarily linked to feelings of isolation and stigma due to their single status. Participants were generally satisfied with the treatment they received from clinics. However, they expressed a desire for more detailed information about potential outcomes from egg freezing and suggested ways in which clinics might address the emotional challenges of undertaking this process as an unpartnered person.

Validation of a modified pre-lysis sample preparation technique for flow cytometric minimal residual disease assessment in multiple myeloma, chronic lymphocytic leukemia, and B-non Hodgkin lymphoma.

BACKGROUND: Minimal residual disease (MRD) assessment of hematopoietic neoplasia below 10-4 requires more leukocytes than is usually attainable by post-lysis preparation. However, not all laboratories are resourced for consensus Euroflow pre-lysis methodology. Our study aim was to validate a modified pre-lysis protocol against our standard post-lysis method for MRD detection of multiple myeloma (MM), chronic lymphocytic leukemia (CLL), and B-non Hodgkin lymphoma (B-NHL), to meet demand for deeper MRD assessment by flow cytometry. METHOD: Clinical samples for MRD assessment of MM, CLL, and B-NHL (50, 30, and 30 cases, respectively) were prepared in parallel by pre and post-lysis methods for the initial validation. Total leukocytes, MRD, and median fluorescence intensity of antigen expression were compared as measures of sensitivity and antigen stability. Lymphocyte and granulocyte composition were compared, assessing relative sample processing stability. Sensitivity of the pre-lysis assay was monitored post validation for a further 18 months. RESULTS: Pre-lysis achieved at least 10-4 sensitivity in 85% MM, 81% CLL, and 90% B-NHL samples versus 24%, 48%, and 26% by post-lysis, respectively, with stable antigen expression and leukocyte composition. Post validation over 18 months with technical expertise improving, pre-lysis permitted 10-5 MRD assessment in 69%, 86%, and 82% of the respective patient groups. CONCLUSION: This modified pre-lysis procedure provides a sensitive, robust, time efficient, and relatively cost-effective alternative for MRD testing by MFC at 10-5 , facilitating clinically meaningful deeper response assessment for MM, CLL, and B-NHL. This method adaptation may facilitate more widespread adoption of highly sensitive flow cytometry-based MRD assessment.

On Ice: The impact of vitrification on the use of eggs in fertility treatment.

The possibility to freeze sperm and embryos has long been available to men and women facing infertility as a result of an illness or medical treatment. However, the ability to successfully cryopreserve human eggs is comparatively recent. The introduction and increasing use of egg vitrification from the mid-2000s onwards, alongside the use of intracytoplasmic sperm injection, has seen improved ongoing clinical pregnancy rates compared with slow freezing methods. Despite concerns, the technology has been widely embraced by the scientific community and in recent years has been applied in a greater variety of contexts. In this short perspective paper, we consider two specific applications for the vitrification of human eggs in routine assisted reproduction practice: social egg freezing and the use of frozen eggs in egg donation. We suggest that vitrification is transforming the reproductive landscape in novel and complex ways and that we must be alert to the challenges, complexities and ethics of such developments, especially for those who may be excluded or marginalised by these techniques.

Running out of time: exploring women's motivations for social egg freezing.

Objective: Few qualitative studies have explored women's use of social egg freezing. Derived from an interview study of 31 participants, this article explores the motivations of women using this technology. Methods: Semi-structured interviews were conducted with 31 users of social egg freezing resident in UK (n = 23), USA (n = 7) and Norway (n = 1). Interviews were face to face (n = 16), through Skype and Facetime (n = 9) or by telephone (n = 6). Data were analyzed using interpretive thematic analysis. Results: Women's use of egg freezing was shaped by fears of running out of time to form a conventional family, difficulties in finding a partner and concerns about "panic partnering", together with a desire to avoid future regrets and blame. For some women, use of egg freezing was influenced by recent fertility or health diagnoses as well as critical life events. A fifth of the participants also disclosed an underlying fertility or health issue as affecting their decision. Conclusion: The study provides new insights in to the complex motivations women have for banking eggs. It identifies how women's use of egg freezing was an attempt to "preserve fertility" in the absence of the particular set of "life conditions" they regarded as crucial for pursuing parenthood. It also demonstrates that few women were motivated by a desire to enhance their career and that the boundaries between egg freezing for medical and for social reasons may be more porous than first anticipated.

Sex Specific Placental Accumulation and Behavioral Effects of Developmental Firemaster 550 Exposure in Wistar Rats.

Firemaster® 550 (FM 550) is a commercial flame retardant mixture of brominated and organophosphate compounds applied to polyurethane foam used in furniture and baby products. Due to widespread human exposure, and structural similarities with known endocrine disruptors, concerns have been raised regarding possible toxicity. We previously reported evidence of sex specific behavioral effects in rats resulting from developmental exposure. The present study expands upon this prior finding by testing for a greater range of behavioral effects, and measuring the accumulation of FM 550 compounds in placental tissue. Wistar rat dams were orally exposed to FM 550 during gestation (0, 300 or 1000 µg/day; GD 9 - 18) for placental measurements or perinatally (0, 100, 300 or 1000 µg/day; GD 9 - PND 21) to assess activity and anxiety-like behaviors. Placental accumulation was dose dependent, and in some cases sex specific, with the brominated components reaching the highest levels. Behavioral changes were predominantly associated with a loss or reversal of sex differences in activity and anxiety-like behaviors. These findings demonstrate that environmental chemicals may sex-dependently accumulate in the placenta. That sex-biased exposure might translate to sex-specific adverse outcomes such as behavioral deficits is a possibility that merits further investigation.

Reproductive technology and the life course: current debates and research in social egg freezing.

While there are currently few confirmed births from previously frozen eggs in the UK, the improved outcomes of new technologies of vitrification and intra-cytoplasmic sperm injection (ICSI) have led to the marketing of egg freezing for non-medical reasons, whereby women are offered the possibility of preserving their eggs until such time as they wish to have a child. Non-academic commentary on this phenomenon has commonly constructed it within a neo-liberal discourse of 'choice', whereby women are positioned as choosing to 'delay motherhood,' often for reasons relating to their careers. However, there have been relatively few research studies which explore either women's awareness and understandings of social egg freezing or the reasons why women consider or undertake egg freezing. This paper summarises the current academic discussion surrounding social egg freezing and outlines the limited body of empirical literature identified from a systematic search of relevant databases. The potential benefits and harms of social egg freezing and the ethical issues it raises are well rehearsed in existing literature, but there is limited empirical evidence about who is accessing social egg freezing, why they are taking this option and what their experiences and future intentions are.

Children travelling for treatment: what we don't know.

Travel for the purposes of obtaining medical treatment is increasing, but very little is known about the extent or nature of child medical travel. This paper discusses the outcome of a systematic search for academic literature on paediatric medical travel, outlines the potential significance of 'return health migration' by parents and children from minority ethnic groups in the UK, and suggests an agenda for future multidisciplinary research in this field.

Effects of seal oil and tuna-fish oil on platelet parameters and plasma lipid levels in healthy subjects.

Fish are a rich source of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), two long-chain polyunsaturated n-3 fatty acids (LC n-3 PUFA) with cardiovascular benefits. A related but less-investigated LC n-3 PUFA, docosapentaenoic acid (DPA), is more common in seal oil and pasture-fed red meats. This study compared indicators of platelet function and plasma lipids in healthy volunteers given supplements containing these different fatty acids (FA) for 14 days. Subjects, randomised into three groups of ten, consumed capsules of tuna oil (210 mg EPA, 30 mg DPA, 810 mg DHA), seal oil (340 mg EPA, 230 mg DPA, 450 mg DHA) or placebo (sunola) oil. Supplementary LC n-3 PUFA levels were approximately 1 g/day in both fish and seal oil groups. Baseline dietary FA and other nutrient intakes were similar in all groups. Both fish and seal oil elevated platelet DHA levels (P < 0.01). Seal oil also raised platelet DPA and EPA levels (P < 0.01), and decreased p-selectin (P = 0.01), a platelet activation marker negatively associated with DPA (P = 0.03) and EPA (P < 0.01) but not DHA. Plasma triacylglycerol decreased (P = 0.03) and HDL-cholesterol levels increased (P = 0.01) with seal oil only. Hence, seal oil may be more efficient than fish oil at promoting healthy plasma lipid profiles and lowering thrombotic risk, possibly due to its high DPA as well as EPA content.