Second generation lethality in RNAseH2a knockout zebrafish.

Removal of ribonucleotides from DNA by RNaseH2 is essential for genome stability, and its impacted function causes the neurodegenerative disease, Aicardi Goutières Syndrome. We have created a zebrafish rnaseh2a mutant to model this process. Surprisingly, RNaseH2a knockouts show little phenotypic abnormality at adulthood in the first generation, unlike mouse knockout models, which are early embryonic lethal. However, the second generation offspring show reduced development, increased ribonucleotide incorporation and upregulation of key inflammatory markers, resulting in both maternal and paternal embryonic lethality. Thus, neither fathers or mothers can generate viable offspring even when crossed to wild-type partners. Despite their survival, rnaseh2a-/- adults show an accumulation of ribonucleotides in both the brain and testes that is not present in early development. Our data suggest that homozygotes possess RNaseH2 independent compensatory mechanisms that are inactive or overwhelmed by the inherited ribonucleotides in their offspring, or that zebrafish have a yet unknown tolerance mechanism. Additionally, we identify ribodysgenesis, the rapid removal of rNMPs and subsequently lethal fragmentation of DNA as responsible for maternal and paternal embryonic lethality.

Advances in Modern Microsurgery.

Background/Objectives: Microsurgery employs techniques requiring optical magnification and specialized instruments to operate on small anatomical structures, including small vessels. These methods are integral to plastic surgery, enabling procedures such as free tissue transfer, nerve reconstruction, replantation, and lymphatic surgery. This paper explores the historical development, advancements, and current applications of microsurgery in plastic surgery. Methods: The databases MEDLINE (via PubMed) and Web of Science were selectively searched with the term "(((microsurgery) OR (advances)) OR (robotic)) OR (AI)) AND (((lymphatic surgery) OR (peripheral nerve surgery)) OR (allotransplantation))" and manually checked for relevance. Additionally, a supplementary search among the references of all publications included was performed. Articles were included that were published in English or German up to June 2024. Results: Modern microsurgical techniques have revolutionized plastic surgery, enabling precise tissue transfers, improved nerve reconstruction, and effective lymphedema treatments. The evolution of robotic-assisted surgery, with systems like da Vinci and MUSA, has enhanced precision and reduced operative times. Innovations in imaging, such as magnetic resonance (MR) lymphography and near-infrared fluorescence, have significantly improved surgical planning and outcomes. Conclusions: The continuous advancements in microsurgery, including supermicrosurgical techniques and robotic assistance, have significantly enhanced the capabilities and outcomes of plastic surgery. Future developments in AI and robotics promise further improvements in precision and efficiency, while new imaging modalities and surgical techniques expand the scope and success of microsurgical interventions.

Development and preliminary validation of a diagnostic prediction model to optimise outpatient management of patients with urolithiasis using urinary stones and intervention quality of life (USIQoL) measure.

PURPOSE: Patients with urinary calculi undergo resource-intensive follow-up. Application of a PROM, Urinary Stones and Intervention Quality of Life (USIQoL), can potentially optimise current practices if it matches the outcomes of traditional follow-up. Our objective was to develop, and conduct, a preliminary validation of the USIQoL based prediction model to aid triage. METHODS: We performed a two phase prospective cohort study. The 1st phase included development of the USIQoL-based decision model using multicentre data. The 2nd phase involved prospective single-blind external validation for the outpatient application. The aim was to evaluate correlations between the USIQoL scores and key predictors; clinical outcomes and global health ratings (EuroQoL EQ-5D). We used statistical analysis to validate USIQoL cut-off scores to aid triage and the decision to intervene. RESULTS: Of 503 patients invited, 91% (n = 455, Development [305] and Validation [150]; M = 308, F = 147) participated. The relationship between USIQoL domain scores and clinical outcomes was consistently significant (estimated odds: PPH 1.24, p < 0.001, 95% CI 1.13-1.36; PSH 1.22, p < 0.001, 95% CI 1.12-1.33). The ROC values for the model were ≥ 0.75. The optimum domain cut-off scores were derived with rising scores implying increased need to intervene. The model demonstrated satisfactory sensitivity (0.81-0.89) and specificity (0.36-0.47). CONCLUSIONS: The study demonstrates satisfactory correlation between the USIQoL and clinical outcomes making this model a valid aid for triage and optimising outpatient management with the cut-off scores able to identify high risk patients who need active treatment.

Extending the Indications for Direct Transcutaneous Lower Blepharoplasty With an Infraorbital Incision to Tear Trough Deformities, Suborbicularis Oculi Fat, Festoons, and Revision Blepharoplasty.

Introduction: Treatment of high-grade lower eyelid deformities with massive skin laxity and retroseptal fat pads can be challenging. Common techniques such as the transconjunctival approach and transcutaneous technique performed through a subciliary incision are associated with increased complication rates. Direct excision of the lower eyelid through an infraorbital incision is an alternative technique that allows safe treatment of dermatochalasis and pigmentation and correction of tear trough deformities, suborbicularis oculi fat, and festoons. This study aimed to determine whether direct transcutaneous blepharoplasty with an infraorbital incision could be indicated for these conditions instead of the established operative methods. Methods: A retrospective study of 21 patients with Fratila grades 9 or 10, Hirmand grade 3, and Barton grade 3 who underwent direct transcutaneous lower eyelid blepharoplasty via an infraorbital incision under local anesthesia was performed. All patients underwent surgery during a 9-year period from January 2010 to December 2018. The follow-up period was 12 months. Results were rated postoperatively using Barton grading. Results: Of 21 consecutive patients (13 women and 8 men), 18 required retroseptal fat pad removal, 3 had laxity of the skin and orbicularis muscle, and 5 had triangular cheek festoons. Combined lower and upper blepharoplasty was performed for 12 patients. All patients were satisfied with their surgical results and major improvements were observed. Scar quality was considered good by all patients. Conclusion: Direct transcutaneous lower blepharoplasty of the orbital rim can be indicated for different tear trough deformities, suborbicularis oculi fat laxity, and festoons.

Introduction: Le traitement des difformités de haut grade de la paupière inférieure avec laxité cutanée majeure et amas de graisse rétroseptale peut être un défi. Les techniques habituelles, comme l'approche transconjonctivale et la technique transcutanée, exécutées au travers d'une incision infraciliare sont associées à des taux de complications plus importants. L'excision directe de la paupière inférieure par une incision sous-orbitaire est une autre technique qui permet un traitement sécuritaire du dermatochalasis, de la pigmentation et de la correction des cernes sombres, le tissu adipeux sous-orbitaire et les poches oculaires. Cette étude visait à déterminer si une blépharoplastie directe transcutanée avec incision sous-orbitaire pouvait recevoir une indication pour ces problèmes à la place des techniques opératoires usuelles. Méthodes: Une étude rétrospective a été menée, incluant 21 patients ayant des grades Fratila 9 ou 10, Hirmand 3 et Barton 3; les patients ont subi sous anesthésie locale une blépharoplastie transcutanée de la paupière inférieure par incision sous-orbitaire. Les interventions ont eu lieu dans une période de 9 ans, de janvier 2010 à décembre 2018. La phase de suivi a été de 12 mois. Les résultats ont été évalués en post opératoire selon l'échelle de Barton. Résultats: Parmi 21 patients consécutifs (13 femmes et 8 hommes), 18 ont nécessité la suppression d'un amas de graisse rétroseptale, 3 avaient une laxité de la peau et du muscle orbiculaire et 5 avaient des poches triangulaires sur les joues. Une blépharoplastie combinée supérieure et inférieure a été pratiquée chez 12 patients. Tous les patients ont été satisfaits par les résultats de la chirurgie et des améliorations majeures ont été constatées. Tous les patients ont considéré que la qualité de la cicatrice était bonne. Conclusion: Une blépharoplastie inférieure transcutanée directe du pourtour orbitaire peut être indiquée pour différentes formes de dépression sombre formant des cernes foncés, la laxité de la graisse oculaire sous-orbitaire et les poches sous les yeux.

Cluster effect for SNP-SNP interaction pairs for predicting complex traits.

Single nucleotide polymorphism (SNP) interactions are the key to improving polygenic risk scores. Previous studies reported several significant SNP-SNP interaction pairs that shared a common SNP to form a cluster, but some identified pairs might be false positives. This study aims to identify factors associated with the cluster effect of false positivity and develop strategies to enhance the accuracy of SNP-SNP interactions. The results showed the cluster effect is a major cause of false-positive findings of SNP-SNP interactions. This cluster effect is due to high correlations between a causal pair and null pairs in a cluster. The clusters with a hub SNP with a significant main effect and a large minor allele frequency (MAF) tended to have a higher false-positive rate. In addition, peripheral null SNPs in a cluster with a small MAF tended to enhance false positivity. We also demonstrated that using the modified significance criterion based on the 3 p-value rules and the bootstrap approach (3pRule + bootstrap) can reduce false positivity and maintain high true positivity. In addition, our results also showed that a pair without a significant main effect tends to have weak or no interaction. This study identified the cluster effect and suggested using the 3pRule + bootstrap approach to enhance SNP-SNP interaction detection accuracy.

Optimal Large Language Model Characteristics to Balance Accuracy and Energy Use for Sustainable Medical Applications.

Background Large language models (LLMs) for medical applications use unknown amounts of energy, which contribute to the overall carbon footprint of the health care system. Purpose To investigate the tradeoffs between accuracy and energy use when using different LLM types and sizes for medical applications. Materials and Methods This retrospective study evaluated five different billion (B)-parameter sizes of two open-source LLMs (Meta's Llama 2, a general-purpose model, and LMSYS Org's Vicuna 1.5, a specialized fine-tuned model) using chest radiograph reports from the National Library of Medicine's Indiana University Chest X-ray Collection. Reports with missing demographic information and missing or blank files were excluded. Models were run on local compute clusters with visual computing graphic processing units. A single-task prompt explained clinical terminology and instructed each model to confirm the presence or absence of each of the 13 CheXpert disease labels. Energy use (in kilowatt-hours) was measured using an open-source tool. Accuracy was assessed with 13 CheXpert reference standard labels for diagnostic findings on chest radiographs, where overall accuracy was the mean of individual accuracies of all 13 labels. Efficiency ratios (accuracy per kilowatt-hour) were calculated for each model type and size. Results A total of 3665 chest radiograph reports were evaluated. The Vicuna 1.5 7B and 13B models had higher efficiency ratios (737.28 and 331.40, respectively) and higher overall labeling accuracy (93.83% [3438.69 of 3665 reports] and 93.65% [3432.38 of 3665 reports], respectively) than that of the Llama 2 models (7B: efficiency ratio of 13.39, accuracy of 7.91% [289.76 of 3665 reports]; 13B: efficiency ratio of 40.90, accuracy of 74.08% [2715.15 of 3665 reports]; 70B: efficiency ratio of 22.30, accuracy of 92.70% [3397.38 of 3665 reports]). Vicuna 1.5 7B had the highest efficiency ratio (737.28 vs 13.39 for Llama 2 7B). The larger Llama 2 70B model used more than seven times the energy of its 7B counterpart (4.16 kWh vs 0.59 kWh) with low overall accuracy, resulting in an efficiency ratio of only 22.30. Conclusion Smaller fine-tuned LLMs were more sustainable than larger general-purpose LLMs, using less energy without compromising accuracy, highlighting the importance of LLM selection for medical applications. © RSNA, 2024 Supplemental material is available for this article.

Multiparameter imaging flow cytometry-based cytokinesis-block micronucleus assay: Reduction of culture time and blood volume for improved efficiency.

In the event of a large-scale incident involving radiological or nuclear exposures, there is a potential for large numbers of individuals to have received doses of radiation sufficient to cause adverse health effects. It is imperative to quickly identify these individuals in order to provide information to the medical community to assist in making decisions about their treatment. The cytokinesis-block micronucleus assay is a well-established method for performing biodosimetry. This assay has previously been adapted to imaging flow cytometry and has been validated as a high-throughput option for providing dose estimates in the range of 0-10 Gy. The goal of this study was to test the ability to further optimize the assay by reducing the time of culture to 48 h from 68 h as well as reducing the volume of blood required for the analysis to 200 µL from 2 mL. These modifications would provide efficiencies in time and ease of processing impacting the ability to manage large numbers of samples and provide dose estimates in a timely manner. Results demonstrated that either the blood volume or the culture time could be reduced while maintaining dose estimates with sufficient accuracy for triage analysis. Reducing both the blood volume and culture time, however, resulted in poor dose estimates. In conclusion, depending on the needs of the scenario, either culture time or the blood volume could be reduced to improve the efficiency of analysis for mass casualty scenarios.

Differential Access to Breast Magnetic Resonance Imaging Compared with Mammography and Ultrasound.

INTRODUCTION: For high-risk women, breast magnetic resonance (MR) is the preferred supplemental imaging option, but spatial access differences may exacerbate disparities in breast care. METHODS: This was a cross-sectional study examining distance between ZIP codes and the nearest breast imaging facility (MR, mammography, ultrasound) using 2023 data from the Food and Drug Administration and the American College of Radiology. Linear regression was used to assess distance differences controlling for Area Deprivation Index (ADI), urbanicity, and population size. Analyses were conducted in 2024. RESULTS: Among the 29,629 ZIP codes with an ADI and known urbanicity, unadjusted mean distance to breast MR was 23.2±25.1 miles (SD) compared with 8.2±8.3 for mammography and 22.2±25.0 for ultrasound. Hence, the average distance to breast MR facilities was 2.8 times further than to mammography facilities. ADI and urbanicity were associated with increased distance to the nearest breast imaging facility. The additional miles associated with the least advantaged areas compared with most advantaged areas was 12.2 (95%CI: 11.3, 13.2) for MR, 11.5 miles (95%CI: 10.6, 12.3) for ultrasound, and 2.4 (95%CI: 2.1, 2.7) for mammography. Compared with metropolitan areas, the additional miles to breast MR facilities was 23.2 (95%CI: 22.5, 24.0) for small/rural areas. CONCLUSIONS: Spatial access is substantially better for mammography sites compared with breast MR or ultrasound sites. Given these findings, consideration of options to mitigate the impact of differential access should be considered. For example, mammography sites could offer contrast-enhanced mammography. Future research should examine the feasibility and effectiveness of this and other options.

Lipoteichoic acid biosynthesis by Staphylococcus aureus is controlled by the MspA protein.

Staphylococcus aureus produces a plethora of virulence factors critical to its ability to establish an infection and cause disease. We have previously characterized a small membrane protein, MspA, which has pleiotropic effects on virulence and contributes to S. aureus pathogenicity in vivo. Here we report that mspA inactivation triggers overaccumulation of the essential cell wall component, lipoteichoic acid (LTA), which, in turn, decreases autolytic activity and leads to increased cell size due to a delay in cell separation. We show that MspA directly interacts with the enzymes involved in LTA biosynthesis (LtaA, LtaS, UgtP, and SpsB), interfering with their normal activities. MspA, in particular, interacts with the type I signal peptidase SpsB, limiting its cleavage of LtaS into its active form. These findings suggest that MspA contributes to maintaining a physiological level of LTA in the cell wall by interacting with and inhibiting the activity of SpsB, thereby uncovering a critical role for the MspA protein in regulating cell envelope biosynthesis and pathogenicity.IMPORTANCEThe S. aureus cell envelope, comprising the cytoplasmic membrane, a thick peptidoglycan layer, and the anionic polymers lipoteichoic acid and wall teichoic acids, is fundamental for bacterial growth and division, as well as being the main interface between the pathogen and the host. It has become increasingly apparent that the synthesis and turnover of cell envelope components also affect the virulence of S. aureus. In this study, we show that MspA, an effector of S. aureus virulence, contributes to the maintenance of normal levels of lipoteichoic acid in the cell wall, with implications on cell cycle and size. These findings further our understanding of the connections between envelope synthesis and pathogenicity and suggest that MspA represents a promising target for the development of future therapeutic strategies.

Tick salivary proteome and lipidome with low glycan content correlate with allergic type reactions in the zebrafish model.

Ticks, as hematophagous ectoparasites, can manipulate host immune and metabolic processes, causing tick-borne allergies such as α-Gal syndrome (AGS). Glycolipids with bound galactose-alpha-1-3-galactose (α-Gal) are potential allergenic molecules associated with AGS. Nevertheless, proteins and lipids lacking α-Gal modifications may contribute to tick salivary allergies and be linked to AGS. In this study, we characterized the effect of deglycosylated tick salivary proteins without lipids on treated zebrafish fed with dog food formulated with mammalian (beef, lamb, pork) meat by quantitative proteomics analysis of intestinal samples. The characterization and functional annotations of tick salivary lipids with low representation of glycolipids was conducted using a lipidomics approach. Results showed a significant effect of treatment with saliva and saliva deglycosylated protein fraction on zebrafish abnormal or no feeding (p < 0.005). Treatment with this fraction affected multiple metabolic pathways, defense responses to pathogens and protein metabolism, which correlated with abnormal or no feeding. Lipidomics analysis identified 23 lipid classes with low representation of glycolipids (0.70% of identified lipids). The lipid class with highest representation was phosphatidylcholine (PC; 26.66%) and for glycolipids it corresponded to diacylglycerol (DG; 0.48%). Qualitative analysis of PC antibodies revealed that individuals bitten by ticks were more likely to produce PC-IgG antibodies (p < 0.001). DG levels were significantly higher in tick salivary glands (p < 0.05) compared with tick saliva and salivary fractions. The α-Gal content was higher in tick saliva than in deglycosylated saliva and lipid fractions. These results support a possible role for tick salivary proteins and lipids without α-Gal modifications in AGS.

Making the most of life: environmental choice during rearing enhances the ability of laying hens to take opportunities.

Introduction: The potential of aviary housing for improving laying hen (Gallus gallus domesticus) welfare will be constrained if rearing conditions limit the hens' behavioral ability to take opportunities. Incorporating theories on developmental plasticity and animal agency, this study aimed to determine: (1) whether a choice of litter and perch types during rearing would promote long-lasting changes in use of novel locations and resources, and (2) the influence of timing of choice provision. Methods: Laying hen chicks were assigned to either a "Single-choice" (one litter and perch type) or "Multi-choice" environment (four litter and perch types) during "Early" (day 1-week 4) and "Late" rearing (week 5-15). The environments were switched in half of the 16 pens in week 5, resulting in a 2 × 2 factorial design with four choice environment by period combinations. The allocation of perch and litter space was the same across all treatment combinations. In week 16, all groups were moved to standard aviary laying pens (Laying period, week 16-27). Results: When first moved to the laying pens, hens with Multi-choice in either or both rearing periods were quicker to spread out in their pen than hens with Single-choice throughout rearing. Multi-choice in Early rearing also reduced the latency to use novel elevated structures (perches and nests) in the laying pens. Multi-choice during Late rearing increased success in finding and consuming hidden mealworms (tested in weeks 9-17) and increased the proportion of eggs laid on elevated nesting trays. Numerically, hens switched from Multi-choice to Single-choice in week 5 used the outdoor range less than hens switched from Single-choice to Multi-choice. Discussion: These results support the hypothesis that offering multiple resource choices during rearing improves hens' ability to make the most of new opportunities by being more proactive in exploring and exploiting newly available resources. In different opportunity challenges, hens showed positive outcomes in response to choice during Early, Late or both stages of rearing, suggesting that best results can be obtained by offering environmental choice throughout rearing.

Internalizing Psychiatric Symptoms in People with Mosaicism for Trisomy 21.

People with mosaicism for trisomy 21 have been shown to exhibit the many of same phenotypic traits present in people with non-mosaic Down syndrome, but with varying symptom severity. However, the behavioral phenotype of people with mosaic Down syndrome (mDS) has not been well characterized. This study aimed to examine the prevalence of self-report and caregiver-report symptoms of depression and anxiety among a sample of 62 participants with mDS aged 12 - 46, and assess their association with the percentage of trisomy 21 in blood and/or buccal mucosa cells. The results showed that 53% of the participants reported clinically significant depression symptoms and 76% reported clinically significant anxiety symptoms. No clear associations were observed between the percentage of trisomic cells and total anxiety or depression, but a significant positive association between the proband-reported specific fears subscale and the percentage of trisomic cells in buccal specimens was detected (r = .43, p = .007). This study highlights the high occurrence of depression and anxiety symptoms in individuals with mDS and the need for routine assessment to optimize their care. It also demonstrates the ability of people with mDS to complete these evaluations, thereby supporting their inclusion in research studies/clinical trials.

Evaluating the delivery of care by telemedicine for incarcerated people living with HIV: a cohort study.

BACKGROUND: The use of telemedicine has grown significantly since the COVID-19 pandemic and has the potential to improve access to specialized care for otherwise underserved populations. Incarcerated people living with HIV (PLWH) could potentially benefit from expanded access to HIV care through telemedicine. METHODS: All PLWH who were incarcerated within the Tennessee Department of Corrections and received care through the HIV telemedicine clinic at Regional One Hospital between 5/1/2019 through 2/28/2022 were identified from the electronic health records (EHR). Demographics, laboratory data, vaccine history, and treatment outcomes were abstracted from the EHR. Retention in care and viral suppression were defined using Centers for Disease Control and Prevention definitions. RESULTS: Of the 283 incarcerated PLWH receiving care from this telemedicine clinic, 78% remained retained in care and 94% achieved or maintaining viral suppression at 12 months. Many preventative care measures remained unperformed or undocumented, including vaccinations and testing for concurrent sexually transmitted infections. There were 56 patients (20%) found to have chronic hepatitis C in this population, with 71% either cured or still on treatment in this study period. CONCLUSIONS: Retention in care and viral suppression rates were excellent among incarcerated PLWH receiving telemedicine care for their HIV. HIV related primary health care screenings and vaccinations, however, were less consistently documented and represent areas for improvement.

Assessing the Correlation between Allostatic Load and False-Positive Image-Guided Breast Biopsies.

Background: Allostatic load (AL) is the accumulation of physiological dysregulation attributed to repeated activation of the stress response over a lifetime. We assessed the utility of AL as a prognostic measure for high-risk benign breast biopsy pathology results. Method: Eligible patients were women 18 years or older, with a false-positive outpatient breast biopsy between January and December 2022 at a tertiary academic health center. AL was calculated using 12 variables representing four physiological systems: cardiovascular (pulse rate, systolic and diastolic blood pressures, total cholesterol, high-density lipoprotein, and low-density lipoprotein); metabolic (body mass index, albumin, and hemoglobin A1C); renal (creatinine and estimated glomerular filtration rate); and immune (white blood cell count). Multivariable logistic regression was used to assess the association between AL before biopsy and breast biopsy outcomes controlling for patients' sociodemographics. Results: In total, 170 women were included (mean age, 54.1 ± 12.9 years): 89.4% had benign and 10.6% had high-risk pathologies (radial scar/complex sclerosing lesion, atypical ductal or lobular hyperplasia, flat epithelial atypia, intraductal papilloma, or lobular carcinoma in-situ). A total of 56.5% were White, 24.7% Asian, and 17.1% other races. A total of 32.5% identified as Hispanic. The mean breast cancer risk score using the Tyrer-Cuzick model was 11.9 ± 7.0. In multivariable analysis, with every one unit increase in AL, the probability of high-risk pathology increased by 37% (odds ratio, 1.37; 95% confidence interval, 1.03, 1.81; p = 0.03). No significant association was seen between high-risk pathology and age, ethnicity, breast cancer risk, or area deprivation index. Conclusion: Our findings support that increased AL, a biological marker of stress, is associated with high-risk pathology among patients with false-positive breast biopsy results.

The Establishment of a Novel In Vitro System for Culturing Cytauxzoon felis.

Cytauxzoonosis, a highly fatal tick-borne disease in domestic cats caused by Cytauxzoon felis, poses diagnostic and therapeutic challenges due to the inability to culture the parasite in vitro. This study aimed to artificially replicate C. felis infection and characterize in vitro replication kinetics. Concanavalin A-activated feline embryonal macrophages (Fcwf-4) were plated at 3-5 × 105 cells/mL and incubated with C. felis-positive blood samples from either a (1) chronically infected bobcat (Lynx rufus), (2) chronically infected domestic cat, or (3) acutely infected domestic cat with clinical signs of cytauxzoonosis. Temporal changes in parasite load were quantified by droplet digital PCR (ddPCR), and the inhibition of infection/replication was assessed using atovaquone, imidocarb dipropionate (ID), artemisinin, ponazuril, and neutralizing antibodies. Tick cell lines AAE2 and ISE6 were also tested for infection. In vitro inoculation with chronic infection led to transient replication, while acute infection resulted in sustained replication beyond 10 days post-inoculation. Atovaquone, ID, and artemisinin inhibited replication, and neutralizing antibodies prevented infection. The inoculation of tick cells in vitro indicated infection; however, parasite replication was not observed. The results of this study established an in vitro model for studying infection dynamics, assessing therapy efficacy, and testing vaccination strategies in cytauxzoonosis-infected cats.

Circulating free insulin-like growth factor-I and prostate cancer: a case-control study nested in the European prospective investigation into cancer and nutrition.

BACKGROUND: Circulating total insulin-like growth factor-I (IGF-I) is an established risk factor for prostate cancer. However, only a small proportion of circulating IGF-I is free or readily dissociable from IGF-binding proteins (its bioavailable form), and few studies have investigated the association of circulating free IGF-I with prostate cancer risk. METHODS: We analyzed data from 767 prostate cancer cases and 767 matched controls nested within the European Prospective Investigation into Cancer and Nutrition cohort, with an average of 14-years (interquartile range = 2.9) follow-up. Matching variables were study center, length of follow-up, age, and time of day and fasting duration at blood collection. Circulating free IGF-I concentration was measured in serum samples collected at recruitment visit (mean age 55 years old; standard deviation = 7.1) using an enzyme-linked immunosorbent assay (ELISA). Conditional logistic regressions were performed to examine the associations of free IGF-I with risk of prostate cancer overall and subdivided by time to diagnosis (≤ 14 and > 14 years), and tumor characteristics. RESULTS: Circulating free IGF-I concentrations (in fourths and as a continuous variable) were not associated with prostate cancer risk overall (odds ratio [OR] = 1.00 per 0.1 nmol/L increment, 95% CI: 0.99, 1.02) or by time to diagnosis, or with prostate cancer subtypes, including tumor stage and histological grade. CONCLUSIONS: Estimated circulating free IGF-I was not associated with prostate cancer risk. Further research may consider other assay methods that estimate bioavailable IGF-I to provide more insight into the well-substantiated association between circulating total IGF-I and subsequent prostate cancer risk.

Identifying proteomic risk factors for overall, aggressive, and early onset prostate cancer using Mendelian Randomisation and tumour spatial transcriptomics.

BACKGROUND: Understanding the role of circulating proteins in prostate cancer risk can reveal key biological pathways and identify novel targets for cancer prevention. METHODS: We investigated the association of 2002 genetically predicted circulating protein levels with risk of prostate cancer overall, and of aggressive and early onset disease, using cis-pQTL Mendelian randomisation (MR) and colocalisation. Findings for proteins with support from both MR, after correction for multiple-testing, and colocalisation were replicated using two independent cancer GWAS, one of European and one of African ancestry. Proteins with evidence of prostate-specific tissue expression were additionally investigated using spatial transcriptomic data in prostate tumour tissue to assess their role in tumour aggressiveness. Finally, we mapped risk proteins to drug and ongoing clinical trials targets. FINDINGS: We identified 20 proteins genetically linked to prostate cancer risk (14 for overall [8 specific], 7 for aggressive [3 specific], and 8 for early onset disease [2 specific]), of which the majority replicated where data were available. Among these were proteins associated with aggressive disease, such as PPA2 [Odds Ratio (OR) per 1 SD increment = 2.13, 95% CI: 1.54-2.93], PYY [OR = 1.87, 95% CI: 1.43-2.44] and PRSS3 [OR = 0.80, 95% CI: 0.73-0.89], and those associated with early onset disease, including EHPB1 [OR = 2.89, 95% CI: 1.99-4.21], POGLUT3 [OR = 0.76, 95% CI: 0.67-0.86] and TPM3 [OR = 0.47, 95% CI: 0.34-0.64]. We confirmed an inverse association of MSMB with prostate cancer overall [OR = 0.81, 95% CI: 0.80-0.82], and also found an inverse association with both aggressive [OR = 0.84, 95% CI: 0.82-0.86] and early onset disease [OR = 0.71, 95% CI: 0.68-0.74]. Using spatial transcriptomics data, we identified MSMB as the genome-wide top-most predictive gene to distinguish benign regions from high grade cancer regions that comparatively had five-fold lower MSMB expression. Additionally, ten proteins that were associated with prostate cancer risk also mapped to existing therapeutic interventions. INTERPRETATION: Our findings emphasise the importance of proteomics for improving our understanding of prostate cancer aetiology and of opportunities for novel therapeutic interventions. Additionally, we demonstrate the added benefit of in-depth functional analyses to triangulate the role of risk proteins in the clinical aggressiveness of prostate tumours. Using these integrated methods, we identify a subset of risk proteins associated with aggressive and early onset disease as priorities for investigation for the future prevention and treatment of prostate cancer. FUNDING: This work was supported by Cancer Research UK (grant no. C8221/A29017).

Effects of Early and Late Time-Restricted Feeding on Parameters of Metabolic Health: An Explorative Literature Assessment.

Chrono-nutrition (meal timing) aligns food consumption with one's circadian rhythm. The first meal (e.g., breakfast) likely promotes synchronization of peripheral circadian clocks, thereby supporting metabolic health. Time-restricted feeding (TRF) has been shown to reduce body weight (BW) and/or improve cardiovascular biomarkers. In this explorative literature assessment, 13 TRF randomized controlled trials (RCTs) were selected from PubMed and Scopus to evaluate the effects of early (eTRF: first meal before 10:30 a.m.) and late TRF (lTRF: first meal after 11:30 a.m.) on parameters of metabolic health. Although distinct variations in study design were evident between reports, TRF consistently decreased energy intake (EI) and BW, and improved insulin resistance as well as systolic blood pressure. eTRF seemed to have a greater beneficial effect than lTRF on insulin resistance (HOMA-IR). Importantly, most studies did not appear to consider chronotype in their evaluation, which may have underestimated TRF effects. TRF intervention may be a promising approach for risk reduction of human metabolic diseases. To conclusively determine benefits of TRF and identify clear differences between eTRF and lTRF, future studies should be longer-term (≥8 weeks) with well-defined (differences in) feeding windows, include participants chronotypically matching the intervention, and compare outcomes to those of control groups without any dietary limitations.

Use of FRET-Sensor 'Mermaid' to Detect Subtle Changes in Membrane Potential of Primary Mouse PASMCs.

Subtle changes in the membrane potential of pulmonary arterial smooth muscle cells (PASMCs) are pivotal for controlling pulmonary vascular tone, e.g., for initiating Hypoxic Pulmonary Vasoconstriction, a vital mechanism of the pulmonary circulation. In our study, we evaluated the ability of the fluorescence resonance energy transfer (FRET)-based voltage-sensor Mermaid to detect such subtle changes in membrane potential. Mouse PASMCs were isolated and transduced with Mermaid-encoding lentiviral vectors before the acceptor/donor emission ratio was assessed via live cell FRET-imaging. Mermaid's sensitivity was tested by applying specific potassium chloride (KCl) concentrations. These KCl concentrations were previously validated by patch clamp recordings to induce depolarization with predefined amplitudes that physiologically occur in PASMCs. Mermaid's emission ratio dose-dependently increased upon depolarization with KCl. However, Mermaid formed unspecific intracellular aggregates, which limited the usefulness of this voltage sensor. When analyzing the membrane rim only to circumvent these unspecific signals, Mermaid was not suitable to resolve subtle changes in the membrane potential of ≤10 mV. In summary, we found Mermaid to be a suitable alternative for reliably detecting qualitative membrane voltage changes of more than 10 mV in primary mouse PASMCs. However, one should be aware of the limitations associated with this voltage sensor.