Key Time Points Research Articles

The Cleft Collective: protocol for a longitudinal prospective cohort study

IntroductionCleft lip and/or palate (CL/P) affects 1 in 700 live births globally. Children born with CL/P and their families face various challenges throughout the child’s development. Extant research is often...

Opioid initiation timing and palliative care referrals in advanced cancer: retrospective study

ObjectivesEarly opioid initiation is recommended for advanced cancer pain, however the timing of opioid commencement in relation to diagnosis has not been described, and the role of palliative care prescribers...

A Switch from Glial to Neuronal Gene Expression Alterations in the Spinal Cord of SIV-infected Macaques on Antiretroviral Therapy.

Despite antiretroviral therapy (ART), HIV-associated peripheral neuropathy remains one of the most prevalent neurologic manifestations of HIV infection. The spinal cord is an essential component of sensory pathways, but spinal cord sampling and evaluation in people with HIV has been very limited, especially in those on ART. The SIV/macaque model allows for assessment of the spinal cord at key time points throughout infection with and without ART. In this study, RNA was isolated from the spinal cord of uninfected, SIV+, and SIV + ART animals to track alterations in gene expression using global RNA-seq.Next, the SeqSeek platform was used to map changes in gene expression to specific cell types. Pathway analysis of differentially expressed genes demonstrated that highly upregulated genes in SIV-infected spinal cord aligned with interferon and viral response pathways. Additionally, this upregulated gene set significantly overlapped with those expressed in myeloid-derived cells including microglia. Downregulated genes were involved in cholesterol and collagen biosynthesis, and TGF-b regulation of extracellular matrix. In contrast, enriched pathways identified in SIV + ART animals included neurotransmitter receptors and post synaptic signaling regulators, and transmission across chemical synapses. SeqSeek analysis showed that upregulated genes were primarily expressed by neurons rather than glia. These findings indicate that pathways activated in the spinal cord of SIV + ART macaques are predominantly involved in neuronal signaling rather than proinflammatory pathways. This study provides the basis for further evaluation of mechanisms of SIV infection + ART within the spinal cord with a focus on therapeutic interventions to maintain synaptodendritic homeostasis.

Single-nucleus chromatin landscape dataset of mouse brain development and aging

The development and aging of the brain constitute a lifelong dynamic process, marked by structural and functional changes that entail highly coordinated cellular differentiation and epigenetic regulatory mechanisms. Chromatin accessibility serves as the foundational basis for genetic activity. However, the holistic and dynamic chromatin landscape that spans various brain regions throughout development and ageing remains predominantly unexplored. In this study, we employed single-nucleus ATAC-seq to generate comprehensive chromatin accessibility maps, incorporating data from 69,178 cells obtained from four distinct brain regions – namely, the olfactory bulb (OB), cerebellum (CB), prefrontal cortex (PFC), and hippocampus (HP) – across key developmental time points at 7 P, 3 M, 12 M, and 18 M. We delineated the distribution of cell types across different age stages and brain regions, providing insight into chromatin accessible regions and key transcription factors specific to different cell types. Our data contribute to understanding the epigenetic basis of the formation of different brain regions, providing a dynamic landscape and comprehensive resource for revealing gene regulatory programs during brain development and aging.

INVESTIGATING THE RELATIONSHIP BETWEEN BLEEDING, CLOTTING, AND COAGULOPATHY DURING AUTOMATED PARTIAL REBOA STRATEGIES IN A HIGHLY LETHAL PORCINE HEMORRHAGE MODEL.

Background: Death due to hemorrhagic shock, particularly, noncompressible truncal hemorrhage, remains one of the leading causes of potentially preventable deaths. Automated partial and intermittent resuscitative endovascular balloon occlusion of the aorta (i.e., pREBOA and iREBOA, respectively) are lifesaving endovascular strategies aimed to achieve quick hemostatic control while mitigating distal ischemia. In iREBOA, the balloon is titrated from full occlusion to no occlusion intermittently, whereas in pREBOA, a partial occlusion is maintained. Therefore, these two interventions impose different hemodynamic conditions, which may impact coagulation and the endothelial glycocalyx layer. In this study, we aimed to characterize the clotting kinetics and coagulopathy associated with iREBOA and pREBOA, using thromboelastography (TEG). We hypothesized that iREBOA would be associated with a more hypercoagulopathic response compared with pREBOA due to more oscillatory flow. Methods: Yorkshire swine (n = 8/group) were subjected to an uncontrolled hemorrhage by liver transection, followed by 90 min of automated pREBOA, iREBOA, or no balloon support (control). Hemodynamic parameters were continuously recorded, and blood samples were serially collected during the experiment (i.e., eight key time points: baseline (BL), T0, T10, T30, T60, T90, T120, T210 min). Citrated kaolin heparinase assays were run on a TEG 5000 (Haemonetics, Niles, IL). General linear mixed models were employed to compare differences in TEG parameters between groups and over time using STATA (v17; College Station, TX), while adjusting for sex and weight. Results: As expected, iREBOA was associated with more oscillations in proximal pressure (and greater magnitudes of peak pressure) because of the intermittent periods of full aortic occlusion and complete balloon deflation, compared to pREBOA. Despite these differences in acute hemodynamics, there were no significant differences in any of the TEG parameters between the iREBOA and pREBOA groups. However, animals in both groups experienced a significant reduction in clotting times (R time: P < 0.001; K time: P < 0.001) and clot strength (MA: P = 0.01; G: P = 0.02) over the duration of the experiment. Conclusions: Despite observing acute differences in peak proximal pressures between the iREBOA and pREBOA groups, we did not observe any significant differences in TEG parameters between iREBOA and pREBOA. The changes in TEG profiles were significant over time, indicating that a severe hemorrhage followed by both pREBOA and iREBOA can result in faster clotting reaction times (i.e., R times). Nevertheless, when considering the significant reduction in transfusion requirements and more stable hemodynamic response in the pREBOA group, there may be some evidence favoring pREBOA usage over iREBOA.

Assessment of breast cancer optimal care timeframes in a regional and rural real-world setting: A regional cancer centre experience in Australia.

e12614 Background: The Breast Cancer Optimal Care Pathway (Cancer Australia) is a national patient-centred guideline that seeks to standardise the management pathway for breast cancer patients and spans from early diagnosis through to survivorship. The guideline provides suggested optimal management-based timeframes including the time from general practitioner (GP) referral to specialist review (&lt;2 weeks), time from specialist review to completion of investigations and multidisciplinary discussion (MDT) (&lt;2 weeks), time to initiation of neoadjuvant therapy from MDT (&lt;4 weeks) and time to surgery following completion of neoadjuvant therapy (&lt;6 weeks). Methods: Retrospective data was obtained for patients undergoing neoadjuvant systemic therapy for newly diagnosed breast cancer between 2021 and 2023 at a regional tertiary cancer centre in Australia (Townsville University Hospital). Data included baseline patient demographics, tumour characteristics, staging, key optimal care pathway timepoints, treatment regimen and outcomes. Median and interquartile range were calculated for key timepoints. Results: Overall, 53 patients (100% female) were identified for inclusion. 10% of patients identified as Indigenous (n=5), 36% lived rurally (n=19) and 13% were treated using a tele-chemotherapy model (n=7). Receptor status was triple-negative in 20 patients (38%), hormone-positive in 17 patients (32%) and HER2-positive in 16 patients (30%). 15% of patients had stage I disease (n=8), 58% had stage II disease (n=31), 21% had stage III disease (n=11), while 6% of patients had oligometastatic stage IV disease at diagnosis (n=3). 24 patients (50%) were seen by a specialist within 2 weeks of GP referral (median: 15 days; IQR 9-19). 46 patients (87%) had a definitive management plan formulated at MDT within 2 weeks of specialist review (median: 5 days; IQR 4-9), with delays most commonly due to pending investigations (n=5; 9%). 48 patients (91%) proceeded with neoadjuvant therapy within 4 weeks of MDT (median: 14 days; IQR 11-22), with delays most commonly due to concurrent infections including COVID. 90% of patients (n=46) were ultimately commenced on neoadjuvant treatment within the 8-week optimal care timeframe from initial GP referral (median: 34 days; IQR 27-47). In comparison to patients living locally, patients residing in rural areas were less likely to achieve this target (79% v 97%, p=0.05). 83% of patients (n=43) underwent surgery within 6-weeks of completion of neoadjuvant treatment (median: 30 days; IQR 27-34). Conclusions: The neoadjuvant breast cancer optimal care timeframes were feasible to achieve in a regional and rural setting. However, further targeted research is needed to improve adherence to optimal care standards for patients from rural areas, such as the timeframe from initial referral to completion of investigations and MDT.

Sex-dependent intra-islet structural rearrangements affecting alpha-to-beta cell interactions lead to adaptive enhancements of Ca2+ dynamics in prediabetic beta cells

Aims/hypothesisPrediabetic pancreatic beta cells can adapt their function to maintain normoglycaemia for a limited period of time, after which diabetes mellitus will manifest upon beta cell exhaustion. Understanding sex-specific beta cell compensatory mechanisms and their failure in prediabetes (impaired glucose tolerance) is crucial for early disease diagnosis and individualised treatment. Our aims were as follows: (1) to determine the key time points of the progression from beta cells’ functional adaptations to their failure in vivo; and (2) to mechanistically explain in vivo sex-specific beta cell compensatory mechanisms and their failure in prediabetes.MethodsIslets from male and female transgenic Ins1CreERT2-GCaMP3 mice were transplanted into the anterior chamber of the eye of 10- to 12-week-old sex-matched C57BL/6J mice. Recipient mice were fed either a control diet (CD) or western diet (WD) for a maximum of 4 months. Metabolic variables were evaluated monthly. Beta cell cytoplasmic free calcium concentration ([Ca2+]i) dynamics were monitored in vivo longitudinally by image fluorescence of the GCaMP3 reporter islets. Global islet beta cell [Ca2+]i dynamics in line with single beta cell [Ca2+]i analysis were used for beta cell coordination studies. The glucagon receptor antagonist L-168,049 (4 mmol/l) was applied topically to the transplanted eyes to evaluate in vivo the effect of glucagon on beta cell [Ca2+]idynamics. Human islets from non-diabetic women and men were cultured for 24 h in either a control medium or high-fat/high-glucose medium in the presence or absence of the glucagon receptor antagonist L-168,049. [Ca2+]i dynamics of human islets were evaluated in vitro after 1 h exposure to Fura-10.ResultsMice fed a WD for 1 month displayed increased beta cell [Ca2+]i dynamics linked to enhanced insulin secretion as a functional compensatory mechanism in prediabetes. Recruitment of inactive beta cells in WD-fed mice explained the improved beta cell function adaptation observed in vivo; this occurred in a sex-specific manner. Mechanistically, this was attributable to an intra-islet structural rearrangement involving alpha cells. These sex-dependent cytoarchitecture reorganisations, observed in both mice and humans, induced enhanced paracrine input from adjacent alpha cells, adjusting the glucose setpoint and amplifying the insulin secretion pathway. When WD feeding was prolonged, female mice maintained the adaptive mechanism due to their intrinsically high proportion of alpha cells. In males, [Ca2+]i dynamics progressively declined subsequent to glucose stimulation while insulin secretion continue to increase, suggesting uncoordinated beta cell function as an early sign of diabetes.Conclusions/interpretationWe identified increased coordination of [Ca2+]i dynamics as a beta cell functional adaptation mechanisms in prediabetes. Importantly, we uncovered the mechanisms by which sex-dependent beta cell [Ca2+]i dynamics coordination is orchestrated by an intra-islet structure reorganisation increasing the paracrine input from alpha cells on beta cell function. Moreover, we identified reduced [Ca2+]i dynamics coordination in response to glucose as an early sign of diabetes preceding beta cell secretory dysfunction, with males being more vulnerable. Alterations in coordination capacity of [Ca2+]i dynamics may thus serve as an early marker for beta cell failure in prediabetes.Graphical

Clinical Care Pathway Time Intervals and Tumor Progression Among Head and Neck Cancer Patients at East Avenue Medical Center Before and During the COVID-19 Pandemic

Objective: To investigate the association between the time intervals of key clinical time points and tumor progression (increase in clinical staging) in head and neck cancer patients before and during the pandemic. Methods: Design: Retrospective Cohort Study Setting: Tertiary Government Training Hospital Participants: A total of 81 head and neck cancer patients who consulted at the OPD and underwent elective surgery between January 1, 2018, and December 31, 2022, under the Department of Otorhinolaryngology – Head and Neck Surgery of East Avenue Medical Center were included in the study; 40 patients comprised the pre-pandemic group and 41 patients-the pandemic group. Results:Majority of patients were men (61.73%), and the mean age was 54 years. The most prevalent tumor site was the oral cavity (37.04%). Most patients were Clinical Stage IV at the time of diagnosis (32.10%) and at the time of surgery (58.02%). In the pre-pandemic period, median time-to-consult was 180 days, time-to-diagnosis was 14 days, and time-to-treatment was 57 days. During the pandemic, median time-to-consult significantly increased to 365 days (Mann-Whitney test, U = 589, p = .028), but time-to-diagnosis decreased to 10 days, and time to-treatment decreased to 43 days, although these were not significant (U = 775, p = .667; U = 809, p = .917). Among the 81 patients in the study, 14 (17.28%) showed tumor progression (pre-pandemic: 6; 15%; pandemic: 8; 19.51%), but there was no significant association between time-to-consult and increase in clinical staging for both pre-pandemic (χ2(38) = 34.2, p = .646) and pandemic groups (χ2(16) = 23.1, p = .110) or between time-to-diagnosis and increase in clinical staging for pre-pandemic (χ2(56) = 36.8, p = .978) and pandemic groups (χ2(23) = 28.3, p = .267). Overall, there was no significant association between time-to-treatment and increase in clinical staging for both pre-pandemic (χ2(62) = 80.00, p = .062) and pandemic groups (χ2(32) = 30.4, p = .548), but a subset of patients with larynx primary tumor site had a statistically significant association between time-to-treatment and tumor progression (χ2(5) = 12.00, p = .035). Conclusion:This study revealed that there was an increase in time to-consult for head and neck cancer patients during the pandemic. However, there was no significant difference in time-to-diagnosis and time-to-treatment. This shows that the Department of ORL-HNS, East Avenue Medical Center has provided pandemic head and neck cancer care similar to before the pandemic. No significant associations were found between tumor progression and time intervals of the key clinical time points but patients who had an increase in clinical stage were noted with longer time-to treatment. It was also observed that more patients were in advanced clinical stages during the pandemic.

Mapping patient education encounters in elective surgery: a cohort study and cross-sectional survey

ObjectiveDevelop a process map of when patients learn about their proposed surgery and what resources patients use to educate themselves.DesignA mixed methods design, combining semistructured stakeholder interviews, quantitative validation using...

#2042 Efficacy and safety of telitacicept in IgA nephropathy: a retrospective study

Abstract Background and Aims Telitacicept is a novel fusion protein composed of a recombinant transmembrane activator, calcium modulator, and cyclophilin ligand interactor (TACI) receptor fused with the fragment crystallizable (Fc) region of human immunoglobulin G (IgG). Telitacicept inhibits the differentiation of immature B cell and survival of long-lived plasma cells by neutralizing the activity of B Lymphocyte Stimulator (BlyS) and A Proliferation-Inducing Ligand (APRIL). It was approved for the treatment of patients with Systemic Lupus Erythematosus (SLE) in China in March 2021. Given that BlyS/APRIL overexpression has also been identified in IgA nephropathy (IgAN), telitacicept is in clinical trials for the treatment in IgAN. In this study, we aim to retrospectively evaluate the efficacy and safety of telitacicept in patients with IgAN. Method In this retrospective study, 27 patients with primary IgAN were enrolled from Nephrology Department of Renmin hospital of Wuhan university in China from February 2022 to March 2023. Patients were administered 160 mg telitacicept by subcutaneous injection based on the previous treatment once a week at least 4 weeks, and were followed up. Adults 20–75 years of age who met the following criteria were included: biopsy-proven primary IgAN and 24-hour proteinuria &amp;gt;0.75 g. Patients were excluded if they had secondary causes of IgAN, 24 h proteinuria &amp;lt;0.75 g or immunocompromised state. Changes in proteinuria, serum creatinine, estimated glomerular filtration rate (eGFR), serum immunoglobulins, complement protein (in blood serum), albumin, urine erythrocyte, blood leukocyte and hemoglobin were analyzed during treatment. Key assessment time points were at 2, 4, 8, 12, 16 and 24 weeks after the first administration. Results In this study, duration of telitacicept exposure ranged from 4 weeks to 32 weeks, the median of which was 16 weeks. After administration of telitacicept, baseline 24 h proteinuria of 3.37 g/d decreased to 1.05 g/d at the Week 24 (z = −4.46, p &amp;lt; 0.05), with reductions statistically significant by Week 4. During telitacicept treatment, the median eGFR showed stable trend without sudden decline. Compared to the baseline eGFR of 68.34 ml/(min 1.73 m2), the eGFR at Week 24 was 72 ml/(min 1.73 m2). Changes in immunoglobulin A (IgA), immunoglobulin G (IgG) and immunoglobulin M (IgM) all showed downward trend while complement protein 3 (C3) and complement protein 4 (C4) maintained stable trend. Specifically, IgA decreased from 2.51 g/L at baseline to 1.41 g/L at Week 24, there was a significant reduction by Week 4 (P = 0.001). The median of IgG decreased from 8.31 g/L at baseline to 6.71 g/L at Week 24, with reductions statistically significant by Week 4 (P = 0.008). IgM decreased significantly at Week 2 from baseline of 1.13 g/L to 1.05 g/L (P = 0.017) and was 0.64 g/L at Week 24. The median C3 was stable from baseline to Week 24 (0.92 g/L, 0.87 g/L, respectively), and C4 were 0.22 g/L and 0.23 g/L, in order. Patients had a significant increase in albumin from baseline to Week 2 (P = 0.009) and the Week 24. (36.7 g/L, 37.1 g/L and 41.8 g/L, respectively). Conversely, there was a continuous reduction in urine erythrocyte, from baseline of 62.46/μL to 7.68/μL at Week 24. There remained stable in blood leukocyte from baseline to Week 24 (8.48 × 109/L, 8.67 × 109/L, respectively), with no significant changes. By the end of the study, 10/27 patients was in complete remission (CR) and 8/27 patients had achieved partial remission (PR). The overall remission rate of primary IgAN with telitacicept treatment reached 67%. Telitacicept was generally well tolerated in patients with IgAN in this study. No treatment-emergent adverse events (AEs) occurred in all 27 patients, such as respiratory tract infection, urinary tract infections and injection site allergic. Conclusion In conclusion, based on the retrospective analysis with a small sample, telitacicept, a novel BLyS/APRIL dual inhibitor, effectively decreases proteinuria in IgAN with stable renal function. In addition, it demonstrates a favorable safety profile in patients. The data are supportive of further study with telitacicept in IgAN.

Transcriptomic and epigenomic insights into pectoral muscle fiber formation at the late embryonic development in pure chicken lines

Long-term intensive genetic selection has led to significant differences between broiler and layer chickens, which are evident during the embryonic period. Despite this, there is a paucity of research on the genetic regulation of the initial formation of muscle fiber morphology in chick embryos. Embryonic d 17 (E17) is the key time point for myoblast fusion completion and muscle fiber morphology formation in chickens. This study aimed to explore the genetic regulatory mechanisms underlying the early muscle fiber morphology establishment in broiler chickens of Cornish (CC) and White Plymouth Rock (RR) and layer chickens of White Leghorn (WW) at E17 using the transcriptomic and chromatin accessibility sequencing of pectoral major muscles. The results showed that broiler chickens exhibited significant higher embryo weight and pectoral major muscle weight at E17 compared to layer chickens (P = 0.000). A total of 1,278, 1,248, and 892 differentially expressed genes (DEGs) of RNA-seq data were identified between CC vs. WW, RR vs. WW, and CC vs. RR, separately. All DEGs were combined for cluster analysis and they were divided into 6 clusters, including cluster 1 with higher expression in broilers and cluster 6 with higher expression in layers. DEGs in cluster 1 were enriched in terms related to macrophage activation (P = 0.002) and defense response to bacteria (P = 0.002), while DEGs in cluster 6 showed enrichment in protein-DNA complex (P = 0.003) and monooxygenase activity (P = 0.000). ATAC-seq data analysis identified a total of 38,603 peaks, with 13,051 peaks for CC, 18,780 peaks for RR, and 6,772 peaks for WW. Integrative analysis of transcriptomic and chromatin accessibility data revealed GOLM1, ISLR2, and TOPAZ1 were commonly upregulated genes in CC and RR. Furthermore, screening of all upregulated DEGs in cluster 1 from CC and RR identified GOLM1, ISLR2, and HNMT genes associated with neuroimmune functions and MYOM3 linked to muscle morphology development, showing significantly elevated expression in broiler chickens compared to layer chickens. These findings suggest active neural system connectivity during the initial formation of muscle fiber morphology in embryonic period, highlighting the early interaction between muscle fiber formation morphology and the nervous system. This study provides novel insights into late chick embryo development and lays a deeper foundation for further research.

Neuroactive steroid hormone trajectories across the menstrual cycle in premenstrual dysphoric disorder (PMDD): the PHASE study.

It is presently not known whether endogenous neuroactive steroid hormone trajectories across the menstrual cycle are distinguishable in women with premenstrual dysphoric disorder (PMDD). To improve the rigor in this area of research, we implemented a validated study methodology, involving blood sample collection at 8 key menstrual cycle timepoints, following which the study data is realigned so that all women are compared at the same biological window (i.e., menstrual cycle subphase). Using liquid chromatography-mass spectrometry (LC-MS), we analyzed serum levels of nine steroid hormones previously implicated in the etiology of PMDD, including allopregnanolone. Other than progesterone (p ≤ 0.001), none of the steroid hormones displayed significant changes across menstrual cycle subphases when comparing participants with PMDD to the healthy controls. A thorough investigation of the progesterone trajectory showed that its left shift in the luteal phase (e.g., earlier rise in progesterone) exposes women with PMDD to a higher periovulatory progesterone and a more acute withdrawal in the late luteal subphase. Results of the present study indicate that the largely overlooked brief periovulatory subphase should be thoroughly examined in PMDD and agree with prior conclusions that rapid progesterone withdrawal associates with the development of negative affect.

Comparison of glenohumeral and scapulothoracic kinematics between fastballs and curveballs during baseball pitching

ABSTRACT Shoulder injuries are common in baseball pitchers and primarily involve the glenohumeral joint. Past analyses have examined shoulder biomechanics during different pitch types simply as the motion of the upper arm relative to the thorax. In this study, glenohumeral and scapulothoracic kinematics were compared between fastballs and curveballs at key timepoints throughout a pitch. Upper extremity kinematics of thirteen collegiate pitchers were collected during fastball and curveball pitches with motion capture. A linear model approach was utilised to estimate scapular kinematics based on measurable humerothoracic motion. Glenohumeral kinematics were computed from the scapular and humeral motion data. Comparisons of scapulothoracic and glenohumeral kinematic variables at times of maximum glenohumeral external rotation, ball release, and maximum glenohumeral internal rotation between pitch types were made using paired t-tests with Benjamini-Hochberg corrections. There were no significant differences in glenohumeral kinematics. Fastballs elicited significantly less scapulothoracic internal rotation and more posterior tilt at maximum glenohumeral external rotation. Fastballs produced significantly less scapulothoracic internal rotation and anterior tilt at maximum glenohumeral internal rotation. This study provides further evidence that risk of injury to the glenohumeral joint may be consistent between fastballs and curveballs and offers insights into subtle differences in scapular kinematics between pitch types.

Evaluation of differential miRNA expression between Fusarium wilt-resistant and -susceptible watermelon varieties

Watermelon (Citrullus lanatus) has been cultivated for nearly one thousand years and remains a commercially important crop. However, continuous planting often leads to the aggravation of many diseases. Watermelon Fusarium wilt (FW) is one of them, which is a fungal soil-borne disease caused by Fusarium oxysporum f. sp. niveum (Fon) that damages plant roots by invading vascular bundles. Therefore, it is imperative to comprehensively characterize the mechanism mediating disease resistance or susceptibility, as well as identify effective resistance genes, in order to breed improved disease-resistant watermelon varieties. miRNAs are endogenous non-coding RNAs that are widely involved in plant growth, development, metabolism, transport, stress response, and pathogen defense. However, there are few reports of disease-associated miRNAs, or their mechanisms of action, in Cucurbitaceae crops. In this study, the roots of both Fon-susceptible and resistant watermelon varieties were infected with GFP-labeled Fon. Three key infection time points were identified. Illumina sequencing was used to obtain Fon-resistance related miRNAs, and degradome sequencing was used to obtain miRNA target genes. A total of 23 differentially expressed miRNAs were identified at the three key infection time points, the degradation group sequencing found their only target gene. The results of this study provide a theoretical basis for studies of miRNA function and regulation during the interaction between Fon and watermelon plants and clues for the screening of watermelon resistance genes. In addition, this information can be used to breed disease-resistant watermelon varieties.

The impact of cohort relationships on BIPOC genetic counseling students: Results from a longitudinal qualitative study.

The transition to graduate school is marked by stress, with academic demands and interpersonal interactions being primary concerns for genetic counseling students. For Black, Indigenous, and People of Color (BIPOC) graduate students, additional stressors caused by the "minority tax" and microaggressions impact their sense of belonging and inclusion. This prospective longitudinal study employed a constructivist grounded theory approach to investigate the experiences of first-year BIPOC genetic counseling students as they transitioned into the first year of their graduate training. We conducted semi-structured interviews with 26 first-year genetic counseling students at three key time points during their first year and analyzed them using reflexive thematic analysis. Here, we report themes related to stressors when transitioning into the genetic counseling training environment, the role of relationships as a source of support in navigating these challenges, and the impact of cohort dynamics on the training experience. Stressors included managing academic rigor and time demands, navigating microaggressions, reactions to discussions about diversity, equity, inclusion, and justice (DEIJ), and managing mental health. Peer relationships emerge as pivotal source of support, but challenging dynamics within the cohort negatively impacted participants, highlighting the importance of fostering an inclusive training environment. Since programs have less control over the composition of each cohort with the advent of the Match system in 2018, we recommend the use of community-building and debriefing activities to strengthen healthy relationships and address problematic dynamics. We recommend that training programs be proactive in creating mentoring relationships between faculty and students rather than waiting until students ask for help. Ultimately, we advocate for a holistic approach to genetic counseling training that maintains academic rigor but also prioritizes the creation of supportive, inclusive, and culturally sensitive learning environments for all students.

Abstract 2564: Circulating monocytes decrease significantly following disease-directed therapy and may reflect disease expansion in Langerhans cell histiocytosis

Abstract Introduction: Langerhans Cell Histiocytosis (LCH) is a rare disorder characterized by clonal hematopoietic cell expansion forming eosinophilic granulomas. Treatment approaches vary based on disease severity. Despite established therapies, assessing treatment response, particularly regarding peripheral blood monocytes, lacks comprehensive markers. Methods: We studied 76 LCH patients, collecting data on demographics, diagnostic methods, and treatment responses. Peripheral blood absolute monocyte counts were analyzed at key time points, including disease onset, progression, and post-therapy reinstatement, using statistical tests for comparison. Results: Analyzing data from 76 patients (average age: 38), we found diverse disease presentations, with common imaging modalities being CT (37), PET (29), and MRI (22). Complete responses were observed in 17 of 49 patients, with 11 undergoing local therapy. Conversely, 5 patients with pulmonary LCH and chronic tobacco use showed persistent progression. Notably, peripheral blood monocyte counts did not differ between initial and latest assessments, but on disease progression, the mean count (0.94 K/µL) significantly exceeded that following therapy reinstatement (0.31, p=0.000794). Results: Analyzing data from 76 patients (average age: 38), we found diverse disease presentations, with common imaging modalities being CT (37), PET (29), and MRI (22). Complete responses were observed in 17 of 49 patients, with 11 undergoing local therapy. Conversely, 5 patients with pulmonary LCH and chronic tobacco use showed persistent progression. Notably, peripheral blood monocyte counts did not differ between initial and latest assessments, but on disease progression, the mean count (0.94 K/µL) significantly exceeded that following therapy reinstatement (0.31, p=0.000794). Discussion: LCH is proposed to result from dysregulated hematopoiesis, leading to sequential somatic mutations during monocyte differentiation leading to clonal expansion of CD1A and CD207+ histiocytes. Our finding of increased monocyte count during disease progression may suggest elevated circulating LCH or heightened monocyte production for subsequent differentiation, potentially contributing to MHC Class 1 upregulation. This phenomena mirrors published findings from our own group in patients with Rosai Dorfman and Erdheim Chester Disease. To further elucidate the molecular underpinnings of LCH and explore the etiology of this monocyte trend, genetic sequencing analysis is in progress. Ultimately, monitoring peripheral blood monocyte dynamics could offer valuable insights into LCH progression and treatment response. Citation Format: Haadi Ali, Sam Reynolds, Sabrina Wilcox, Naina Chipalkatti, Asra Ahmed. Circulating monocytes decrease significantly following disease-directed therapy and may reflect disease expansion in Langerhans cell histiocytosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2564.

Abstract 69: Trajectory of Cardiovascular Health Across Childhood and Adolescence in Project Viva

Introduction: The American Heart Association recently put forth the Life’s Essential 8 (LE8) algorithm to assess cardiovascular health (CVH) status based on 8 health and behavioral factors. Few studies, however, have characterized the natural history of CVH status across development or identified sociodemographic determinants of CVH trajectories in children. We sought to address this research gap using data from Project Viva, a pre-birth cohort in eastern Massachusetts. Methods: We applied the LE8 algorithm to derive CVH score (range 0-100 points) in early childhood (median age 3.2y), mid-childhood (7.7y), early adolescence (13y), and late adolescence (17.5y). We used segmented mixed-effect models to characterize sex-specific CVH trajectories across these life stages and estimate three subject-specific trajectory parameters: timing of inflection point when CVH score declines; slope before and after the inflection point. We used linear regression models to assess the association of child race and ethnicity, mother’s education level, and annual household income with each trajectory parameter. Results: Among 1,523 children included, 48.7% were female, 3.5% Asian, 15.2% Black, 4.8% Hispanic, 11.5% Other, and 65.0% White. The mean (SD) CVH score was 82.6 (8.6) points in early childhood, 84.1 (8.3) points in mid-childhood, 82.0 (9.8) points in early adolescence, and 73.8 (11.5) points in late adolescence. Females (vs. males) exhibited slower rate of CVH score gain before the inflection point (0.3 vs. 1.0 points/y), later timing of inflection point when CVH score started declining (10.1 vs. 9.6y), and slower rate of CVH score decline after the inflection point (-1.3 vs. -1.5 points/y). Children of Black (β 0.48y; 95% CI 0.28, 0.68) or Other (β 0.28y; 95% CI 0.08, 0.49) race had a later timing of inflection point compared with White children. Children whose mothers were college-educated (vs. non-college educated) had higher CVH score throughout childhood, and exhibited faster rate of CVH score gain before the inflection point (β 0.19 points/y; 95% CI 0.03, 0.34), earlier timing of inflection point (β -0.37y; 95% CI -0.53, -0.22), and faster rate of CVH score decline after the inflection point (β -0.24 points/y; 95% CI -0.38, -0.10). No associations were observed for household income. Conclusions: This cohort study identified a key timepoint between mid-childhood and early adolescence where CVH trajectory declines over time, as well as sociodemographic differences in CVH trajectory parameters. Future studies are warranted to assess other determinants and consequences of these trajectory parameters.

Ascorbic acid releases dormancy and promotes germination by an integrated regulation of abscisic acid and gibberellin in Pyrus betulifolia seeds.

Seed dormancy is an important life history state in which intact viable seeds delay or prevent germination under suitable conditions. Ascorbic acid (AsA) acts as a small molecule antioxidant, and breaking seed dormancy and promoting subsequent growth are among its numerous functions. In this study, a germination test using Pyrus betulifolia seeds treated with exogenous AsA or AsA synthesis inhibitor lycorine (Lyc) and water absorption was conducted. The results indicated that AsA released dormancy and increased germination and 20 mmol L-1 AsA promoted cell division, whereas Lyc reduced germination. Seed germination showed typical three phases of water absorption; and seeds at five key time points were sampled for transcriptome analysis. It revealed that multiple pathways were involved in breaking dormancy and promoting germination through transcriptome data, and 12 differentially expressed genes (DEGs) related to the metabolism and signal transduction of abscisic acid (ABA) and gibberellins (GA) were verified by subsequent RT-qPCR. For metabolites, exogenous AsA increased endogenous AsA and GA3 but reduced ABA and the ABA/GA3 ratio. In addition, three genes regulating ABA synthesis were downregulated by AsA, while five genes mediating ABA degradation were upregulated. Taken together, AsA regulates the pathways associated with ABA and GA synthesis, catalysis, and signal transduction, with subsequent reduction in ABA and increase in GA and further the balance of ABA/GA, ultimately releasing dormancy and promoting germination.

Critical Appraisal of Medical System Performance for STEMI Management – a Comprehensive Analysis of Time Efficiency

Abstract Background Time intervals related to ST-segment myocardial infarction (STEMI) revascularization are central determinants for patient outcomes. The current capability of the Romanian STEMI program to meet guideline-recommended time intervals is largely unknown. Aims The present study aims to assess the ability of a regional STEMI network to obtain guideline-recommended time intervals for primary percutaneous coronary intervention (pPCI) and to measure the occurrence and the extent of time delays. Materials and Methods This prospective study included 500 consecutive patients with STEMI at the “Prof. Dr. C.C. Iliescu” Emergency Institute for Cardiovascular Diseases, Bucharest, Romania during a period of 14 months. Complete ischemic timelines were created using several key timepoints. Results A secondary route (transfer from another hospital) was noted in most cases. The main time intervals were the following: patient delay 209 min, emergency medical system delay 66 min, and PCI center delay at 70 min, totaling an ischemic time of 6.4 h. A provisional stop at another hospital involved the addition of 113 min (1.8 h) until STEMI diagnosis and an additional 83 min (1.3 h) from diagnosis to revascularization, totaling a supplementary ischemic time of 3.1 h. In total, 41.5% of the patients were revascularized between 2 and 6 h from symptoms onset. The objective of revascularization in less than 120 min (from first medical contact) was accomplished in 35.5% of the patients. Prehospital thrombolysis was performed in 6.4% of the cases, although its potential benefits could have been expected in 64.5% of the patients. Conclusions Patients with STEMI arrive predominantly via secondary routes to the PCI center, which implies significantly increased ischemic times. The ambulance alert system and primary routes represent by far the most efficient, albeit still imperfect methods of pre-hospital approach. Prehospital thrombolysis did not compensate for the gaps existing in the performance of the current system.

Optimizing Cold Chain Distribution Routes Considering Dynamic Demand: A Low-Emission Perspective

Cold chain logistics, with its high carbon emissions and energy consumption, contradicts the current advocacy for a “low-carbon economy”. Additionally, in the real delivery process, customers often generate dynamic demand, which has the characteristic of being sudden. Therefore, to help cold chain distribution companies achieve energy-saving and emission-reduction goals while also being able to respond quickly to customer needs, this article starts from a low-carbon perspective and constructs a two-stage vehicle distribution route optimization model that minimizes transportation costs and refrigeration costs, alongside carbon emissions costs. This research serves to minimize the above-mentioned costs while also ensuring a quick response to customer demands and achieving the goals of energy conservation and emission reduction. During the static stage, in order to determine the vehicle distribution scheme, an enhanced genetic algorithm is adopted. During the dynamic optimization stage, a strategy of updating key time points is employed to address the dynamic demand from customers. By comparing the dynamic optimization strategy with the strategy of dispatching additional vehicles, it is demonstrated that the presented model is capable of achieving an overall cost reduction of approximately 17.13%. Notably, carbon emission costs can be reduced by around 17.11%. This demonstrates that the dynamic optimization strategy effectively reduces the usage of distribution vehicles and lowers distribution costs.