Regulatory T (Treg) cells induce an immunosuppressive microenvironment that is a major obstacle for successful tumor immunotherapy. Dissecting the regulatory mechanisms between energy metabolism and ...functionality in Treg cells will provide insight toward developing novel immunotherapies against cancer. Here we report that human naturally occurring and tumor-associated Treg cells exhibit distinct metabolic profiles with selectivity for glucose metabolism compared with effector T cells. Treg-mediated accelerated glucose consumption induces cellular senescence and suppression of responder T cells through cross-talk. TLR8 signaling selectively inhibits glucose uptake and glycolysis in human Treg cells, resulting in reversal of Treg suppression. Importantly, TLR8 signaling-mediated reprogramming of glucose metabolism and function in human Treg cells can enhance anti-tumor immunity in vivo in a melanoma adoptive transfer T cell therapy model. Our studies identify mechanistic links between innate signaling and metabolic regulation of human Treg suppression, which may be used as a strategy to advance tumor immunotherapy.
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•Human Tregs exhibit distinct metabolic profiles compared with effector T cells•Activated Tregs uniquely accelerate glucose consumption for their suppression•TLR8 signaling selectively inhibits glucose uptake and glycolysis in human Tregs•TLR8-mediated reprogramming of Treg glucose metabolism enhances anti-tumor immunity
Treg-mediated immunosuppression is a significant obstacle for tumor immunotherapy. Li et al. show that human Treg cells heighten glucose consumption and subsequently trigger cellular senescence and suppression of effector T cells. With TLR8 signaling, they selectively inhibit glucose uptake and glycolysis in human Treg cells, resulting in Treg function reversal and enhanced anti-tumor immunity.
Anatomy Nights is an international public engagement event created to bring anatomy and anatomists back to public spaces with the goal of increasing the public's understanding of their own anatomy by ...comparison with non-human tissues. The event consists of a 30-minute mini-lecture on the anatomy of a specific anatomical organ followed by a dissection of animal tissues to demonstrate the same organ anatomy. Before and after the lecture and dissection, participants complete research surveys designed to assess prior knowledge and knowledge gained as a result of participation in the event, respectively. This study reports the results of Anatomy Nights brain events held at four different venues in the UK and USA in 2018 and 2019. Two general questions were asked of the data: 1) Do participant post-event test scores differ from pre-event scores; and 2) Are there differences in participant scores based on location, educational background, and career. We addressed these questions using a combination of generalized linear models (R's glm function; R version 4.1.0 R Core Team, 2014) that assumed a binomial distribution and implemented a logit link function, as well as likelihood estimates to compare models. Survey data from 91 participants indicate that scores improve on post-event tests compared to pre-event tests, and these results hold irrespective of location, educational background, and career. In the pre-event tests, participants performed well on naming structures with an English name (frontal lobe and brainstem), and showed signs of improvement on other anatomical names in the post-test. Despite this improvement in knowledge, we found no evidence that participation in Anatomy Nights improved participants' ability to apply this knowledge to neuroanatomical contexts (e.g., stroke).
Abstract
STUDY QUESTION
Do the attachment-related dimensions Anxiety and Avoidance and perceived partner and social support in recipients and donors influence disclosure to others about their ...involvement in donor-assisted conception (DAC)?
SUMMARY ANSWER
A higher global score on attachment Avoidance was associated with greater non-disclosure about involvement in DAC by participants to relationship-specific others.
WHAT IS KNOWN ALREADY
Within the context of DAC, the topic of disclosure has been investigated in terms of the ‘if’, ‘when’, and the ‘how’ to disclose about circumstances of conception. Less focus, however, has been directed to investigating psychological theoretical frameworks that influence disclosure decisions to others, i.e. to whom information is disclosed and to what extent details are transparently revealed about the donor programme.
STUDY DESIGN, SIZE, DURATION
The study was of a cross-sectional design and utilized a sample of 301 participants who were, or had been, involved in DAC, and were recruited across states of Australia. An online self-report questionnaire was completed between June 2014 and June 2017.
PARTICIPANTS/MATERIALS, SETTING, METHODS
English speaking participants consisting of 209 female recipients and 92 donors (36 sperm; 48 egg; 8 embryo donors). Of the recipients, 104 had successfully conceived children via donated gametes (68 sperm, 23 eggs, 6 embryos, and 7 recipients where both gametes were donated from 2 donors to create the embryo). Participants anonymously completed an online questionnaire consisting of five sections: Demographics, Donor Conception and Disclosure Practices, the Experiences in Close Relationships-Relationships Structure, the Quality of Relationship Inventory, and the Multidimensional Scale of Perceived Social Support. Pearson correlations, independent samples t-tests, Chi-square, and ANOVA were used to explore the association between attachment Anxiety and Avoidance scores and disclosure about involvement in a DAC programme to significant others (i.e. parents, siblings, in-laws, and friends).
MAIN RESULTS AND THE ROLE OF CHANCE
Compared to published community cohort data, participants reported lower global scores on attachment Anxiety and Avoidance and high levels of romantic partner and social network support, suggestive of secure relationships in the overall study sample. A higher score on attachment Avoidance was associated with less disclosure to significant others in their social network (i.e. parents, siblings, in-laws, and close friends), even in the presence of strong partner support (partial r = −0.248, P = 0.005). Higher scores on attachment Avoidance were inversely associated with level of perceived partner and social network support (all P < 0.05). Irrespective of attachment scores, more than 90% of all participants agreed that a child born of DAC should be told about mode of conception.
LIMITATIONS, REASONS FOR CAUTION
This study utilized a cross-sectional design precluding causal inferences between dimensions of insecure attachment and disclosure practices. Participants were required to self-report on the quality of their relationships with the potential for social desirability respondent bias. The study’s self-selecting sample may limit generalization to participants who were dis-inclined to participate. Specifically, respondents who have an Avoidant attachment style, may have elected not to participate in the study.
WIDER IMPLICATIONS OF THE FINDINGS
Given the increased use of biotechnology and digital facial recognition enabling self-discovery of the donor and the donor’s extended family, non-disclosure about involvement in DAC may have consequences. An ‘Avoidant’ attachment style is important to assess as a potential risk factor for non-disclosure about involvement in DAC across different relational contexts (e.g. close family members and friends). Fertility counsellors should consider introducing a measure of attachment screening as a pre-emptive psychoeducational strategy during donor implications counselling. This information could be used to offer patients insight into concerns they have about DAC disclosures to key important relationships, providing a target of clinical intervention.
STUDY FUNDING/COMPETING INTEREST(S)
No external funds were sought for this work. None of the authors have any competing interests to declare.
TRIAL REGISTRATION NUMBER
N/A.
Liquid chromatography coupled to mass spectrometry (LC-MS) is one of the most powerful tools in identifying and quantitating molecular species. Decreasing column diameter from the millimeter to ...micrometer scale is now a well-developed method which allows for sample limited analysis. Specific fabrication of capillary columns is required for proper implementation and optimization when working in the nanoflow regime. Coupling the capillary column to the mass spectrometer for electrospray ionization (ESI) requires reduction of the subsequent emitter tip. Reduction of column diameter to capillary scale can produce improved chromatographic efficiency and the reduction of emitter tip size increased sensitivity of the electrospray process. This improved sensitivity and ionization efficiency is valuable in analysis of precious biological samples where analytes vary in size, ion affinity, and concentration. In this review we will discuss common approaches and challenges in implementing nLC-MS methods and how the advantages can be leveraged to investigate a wide range of biomolecules.
To gauge fertility specialists' knowledge, clinical practices, and training needs in regard to eating disorders.
Cross-sectional study.
Fertility clinics.
Eighty Australian and New Zealand fertility ...specialists who were members of the Fertility Society of Australia.
None.
Responses to an anonymously completed online questionnaire.
Approximately 54% of doctors correctly identified the body mass index relevant to anorexia nervosa, and 30% identified menstrual disturbances for anorexia, while 63.8% of doctors incorrectly nominated maladaptive weight control behaviors as a characteristic of binge eating disorder. While clinicians (83.7%) agreed it was important to screen for eating disorders during preconception assessments, 35% routinely screened for eating disorders and 8.8% indicated that their clinics had clinical practice guidelines for management of eating disorders. A minority of participants (13.8%) felt satisfied with their level of university training in eating disorders, 37.5% of doctors felt confident in their ability to recognize symptoms of an eating disorder, and 96.2% indicated a need for further education and clinical guidelines. On most items examined, knowledge and clinical practices regarding eating disorders did not differ according to doctor gender or years of clinical experience working as a fertility specialist.
Knowledge about eating disorders in the context of fertility treatment is important. This study highlights the uncertainty among fertility specialists in detecting features of eating disorders. The findings point to the importance of further education and training, including the development of clinical guidelines specific to fertility health care providers.
The circle of Willis (CoW) is an anastomotic arterial network located on the base of the brain. Studies have shown that it demonstrates considerable anatomical variation in humans. This systematic ...review aimed to identify and catalogue the described anatomical variations of the CoW in humans to create a new, comprehensive variation classification system. An electronic literature search of five databases identified 5899 studies. A two‐phase screening process was performed, and studies underwent quality assessment. A total of 42 studies were included in the review. Data were extracted and circles were reconstructed digitally using graphics software. The classification system contains 82 CoW variations in five continuous groups. Group one contains 24 circles with one or more hypoplastic segments only. Group two contains 11 circles with one or more absent segments only. Group three contains 6 circles with hypoplastic and absent segments only. Group four contains 26 circles with one or more accessory segments. Group five contains 15 circles with other types of anatomical variation. Within each group, circles were subcategorised according to the number or type of segments affected. An original coding system was created to simplify the description of anatomical variations of the CoW. The new classification system provides a comprehensive ontology of the described anatomical variations of the CoW in humans. When used with the coding system, it allows the description and categorisation of recorded and unrecorded variants identified in past and future studies. It is applicable to current clinical practice and the anatomical community, including human anatomy education and research.
The classification system provides a comprehensive ontology of the described anatomical variations of the circle of Willis in humans. Group one contains 24 circles with one or more hypoplastic segments only. Group two contains 11 circles with one or more absent segments only. Group three contains six circles with hypoplastic and absent segments only. Group four contains 26 circles with one or more accessory segments. Group five contains 15 circles with other types of anatomical variation.
Background/Objectives: Semen cryopreservation is routinely performed in fertility clinics for a variety of reasons, including fertility preservation and storage of donor sperm, yet the freeze–thaw ...process leads to cellular damage via ice crystal formation, osmotic shock, and supraphysiological levels of oxidative stress. Sperm resistance to damage during the freeze–thaw process varies widely, yet the intrinsic factors associated with sperm cryotolerance are largely unknown. The study aimed to investigate whether poor chromatin condensation renders sperm vulnerable to DNA fragmentation and cell death induced by the freeze–thaw process. Methods: Participants (n = 51) from the general community who met the inclusion criteria collected a semen sample after 3–8 days of abstinence. Neat semen samples underwent traditional semen analysis, aniline blue (AB)-eosin staining for chromatin condensation, the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay for DNA fragmentation, and the Annexin V assay for apoptosis/necrosis, prior to being cryopreserved using the liquid nitrogen vapour method and stored at −196 °C. Stored samples were later thawed at room temperature and processed using density gradient centrifugation. Motile sperm concentration, DNA fragmentation and apoptosis/necrosis were analysed in post-thaw samples. Results: As indicated by a significant interaction effect in linear mixed models, an increased proportion of AB-positive sperm in the pre-freeze sample exacerbated the adverse effect of freezing on sperm DNA fragmentation (p = 0.004), late apoptosis (p = 0.007), and necrosis (p = 0.007). AB-staining was positively correlated with all three parameters in the post-thaw sample (all rs ≥ 0.424, all p < 0.01) and remained significant after adjusting for neat sperm concentration (all partial rs ≥ 0.493, all p < 0.01). Similarly, AB-staining was significantly correlated with the percentage point change in sperm DNA fragmentation (rs = 0.366, p = 0.014) and necrosis (rs = 0.403, p = 0.009), both of which remained significant after adjusting for neat sperm concentration (both partial rs ≥ 0.404, both p < 0.01), and borderline significantly correlated with percentage point change in late apoptosis (rs = 0.307, p = 0.051). Conclusions: Sperm with poorly condensed chromatin may be more susceptible to cellular damage during the freeze–thaw process, independent of pre-freeze sperm concentration. These findings may help to explain the intrinsic variation in sperm resistance to cryodamage within and between individuals that is poorly understood.
The immune mechanisms mediating COVID-19 vaccine attenuation of COVID-19 remain undescribed. We conducted comprehensive analyses detailing immune responses to SARS-CoV-2 virus in blood ...post-vaccination with ChAdOx1 nCoV-19 or a placebo. Samples from randomised placebo-controlled trials (NCT04324606 and NCT04400838) were taken at baseline, onset of COVID-19-like symptoms, and 7 days later, confirming COVID-19 using nucleic amplification test (NAAT test) via real-time PCR (RT-PCR). Serum cytokines were measured with multiplexed immunoassays. The transcriptome was analysed with long, short and small RNA sequencing. We found attenuation of RNA inflammatory signatures in ChAdOx1 nCoV-19 compared with placebo vaccinees and reduced levels of serum proteins associated with COVID-19 severity. KREMEN1, a putative alternative SARS-CoV-2 receptor, was downregulated in placebo compared with ChAdOx1 nCoV-19 vaccinees. Vaccination ameliorates reductions in cell counts across leukocyte populations and platelets noted at COVID-19 onset, without inducing potentially deleterious Th2-skewed immune responses. Multi-omics integration links a global reduction in miRNA expression at COVID-19 onset to increased pro-inflammatory responses at the mRNA level. This study reveals insights into the role of COVID-19 vaccines in mitigating disease severity by abrogating pro-inflammatory responses associated with severe COVID-19, affirming vaccine-mediated benefit in breakthrough infection, and highlighting the importance of clinically relevant endpoints in vaccine evaluation.
Multiple Sclerosis (MS) is an inflammatory and neurodegenerative disease of the central nervous system. The inflammatory process in MS is driven by both T and B cells and current therapies are ...targeted to each of these cell types. Epigenetic mechanisms may provide a valuable link between genes and environment. DNA methylation is the best studied epigenetic mechanism and is recognized as a potential contributor to MS risk. The objective of this study was to identify DNA methylation changes associated with MS in CD19
B-cells. We performed an epigenome-wide association analysis of DNA methylation in the CD19
B-cells from 24 patients with relapsing-remitting MS on various treatments and 24 healthy controls using Illumina 450 K arrays. A large differentially methylated region (DMR) was observed at the lymphotoxin alpha (LTA) locus. This region was hypermethylated and contains 19 differentially methylated positions (DMPs) spanning 860 bp, all of which are located within the transcriptional start site. We also observed smaller DMRs at 4 MS-associated genes: SLC44A2, LTBR, CARD11 and CXCR5. These preliminary findings suggest that B-cell specific DNA-methylation may be associated with MS risk or response to therapy, specifically at the LTA locus. Development of B-cell specific epigenetic therapies is an attractive new avenue of research in MS treatment. Further studies are now required to validate these findings and understand their functional significance.
Racial and ethnic disparities have been well-documented in the outcomes for chronic limb threatening ischemia (CLTI). One purported explanation has been the disease severity at presentation. We ...hypothesized that the disparities in major adverse limb events (MALE) after peripheral vascular intervention (PVI) for CLTI would persist despite controlling for disease severity at presentation using the WIfI (Wound, Ischemia, foot Infection) stage.
The Vascular Quality Initiative PVI dataset (2016-2021) was queried for CLTI. Patients were excluded if they were missing the WIfI stage. The primary end point was the incidence of 1-year MALE, defined as major amputation (through the tibia or fibula or more proximally) or reintervention (endovascular or surgical) of the initial treatment limb. A multivariate hierarchical Fine-Gray analysis was performed, controlling for hospital variation, competing risk of death, and presenting WIfI stage, to assess the independent association of Black/African American race and Latinx/Hispanic ethnicity with MALE. A Cox proportional hazard regression model was used for the 1-year survival analysis.
Overall, 47,830 patients (60%) had had WIfI scores reported (73% White, 20% Black, and 7% Latinx). The 1-year unadjusted cumulative incidence of MALE was 13.1% (95% confidence interval CI, 12.6%-13.5%) for White, 14.3% (95% CI, 13.5%-15.3%) for Black, and 17.0% (95% CI, 15.3%-18.9%) for Latinx patients. On bivariate analysis, the occurrence of MALE was significantly associated with younger age, Black race, Latinx ethnicity, coronary artery disease, cerebrovascular disease, congestive heart failure, hypertension, diabetes, dialysis, intervention level, any prior minor or major amputation, and WIfI stage (P < .001). The cumulative incidence of 1-year MALE increased by increasing WIfI stage: stage 1, 11.7% (95% CI, 10.9%-12.4%); stage 2, 12.4% (95% CI, 11.8%-13.0%); stage 3, 14.8% (95% CI, 13.8%-15.8%); and stage 4, 15.4% (95% CI, 14.3%-16.6%). The cumulative incidence also increased by intervention level: inflow, 10.7% (95% CI, 9.8%-11.7%), femoropopliteal, 12.3% (95% CI, 11.7%-12.9%); and infrapopliteal, 14.1% (95% CI, 13.5%-14.8%). After adjustment for WIfI stage only, Black race (subdistribution hazard ratio SHR, 1.30; 95% CI, 1.17-1.44; P < .001) and Latinx ethnicity (SHR, 1.58; 95% CI, 1.37-1.81; P < .001) were associated with an increased 1-year hazard of MALE compared with White race. On adjusted multivariable analysis, MALE disparities persisted for Black/African American race (SHR, 1.12; 95% CI, 1.01-1.25; P = .028) and Latinx/Hispanic ethnicity (SHR, 1.34; 95% CI, 1.16-1.54; P < .001) compared with White race.
Black/African American and Latinx/Hispanic patients had a higher associated hazard of MALE after PVI for CLTI compared with White patients despite an adjustment for WIfI stage at presentation. These results suggest that disease severity at presentation does not account for disparities in outcomes. Further work should focus on better understanding the underlying mechanisms for disparities in historically marginalized racial and ethnic groups presenting with CLTI.
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