Serial Collection Research Articles

2-Deoxyglucose and hydroxychloroquine HPLC-MS-MS analytical methods and pharmacokinetic interactions after oralco-administration in male rats.

Our previous work has shown a synergistic tumoricidal efficacy of combining the hexokinase (HK) inhibitor 2-deoxyglucose (2-DG) and the autophagy inhibitor chloroquine (CQ) through intraperitoneal injections on HK2-addicted prostate cancers in animal models. The pharmacokinetic (PK) behaviors of these oral drugs after simultaneous oral administration have not been reported. We developed high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) analytical methods for 2-DG and the clinically favored drug hydroxychloroquine (HCQ) for sera samples. Using a jugular vein-cannulated male rat model with serial blood collection before and after a single gavage dose of each drug alone or in combination, we examined their PK metrics for drug-drug interactions. The data demonstrated a rapid and complete separation of 2-DG from common monosaccharides by HPLC-MS-MS multi-reaction monitoring. Application of the HPLC-MS-MS 2-DG and HCQ methods to sera samples of nine rats showed a peak time (Tmax ) for 2-DG of 0.5 h after 2-DG alone or with HCQ and identical post-peak half-life of approximately 1 h. With a seemingly bi-modal time course for HCQ, the Tmax for HCQ alone (1.2 h) was faster than that for the combination (2 h; p = .017). After combination dosing, the peak concentration (Cmax ) and area under the curve (AUC0-4h ) of 2-DG were decreased by 53.8% (p = .0004) and 53.7% (p = .0001), whereas AUC0-8h for HCQ was decreased by 30.8% (p = .0279) from the respective single dosing. Without changing the mean residence time (MRT0-∞ ) of each drug, the combination affected the apparent volume of distribution (Vd ) and clearance (CL) of 2-DG, and CL for HCQ without affecting its Vd . We observed significant negative PK interactions, probably at the intestinal absorption level, between 2-DG and HCQ taken simultaneously by mouth. Future optimization efforts are warranted for their combination regimen for clinical translation.

Information Sources Preferred by Students in Academic Writing

The study was conducted to directly determine the information resources used by students. Also, the study specifically aimed to identify which format and type of information resources are preferred by students when writing their researches or assignments. A devised structured survey questionnaire; other parts of the tool were patterned with the study of Dilevko & Gottlieb5. The survey was administered 1st Semester of 2022-2023 to eighty (n= 80) students, forty (40) each from second and fifth year plus students. The collected surveys were sorted and tabulated.Mean, percentage and rank were used to treat the data statistically. Results demonstrated that despite the proliferation of online information sources, students still preferred to use print when writing their researches or assignments.In terms of types, regardless of format, books were still preferred over journals and magazines. This result can be attributed to the library’s weak Serials (journals, magazines, etc.) collections. The library should consider to balance and strengthen its collections in both format and type.

Higher Molecular Injury at Diagnosis of Acute Cellular Rejection Increases the Risk of Lung Allograft Failure: A Clinical Trial.

Rationale: The association of acute cellular rejection (ACR) with chronic lung allograft dysfunction (CLAD) in lung transplant recipients has primarily been described before consensus recommendations incorporating restrictive phenotypes. Furthermore, the association of the degree of molecular allograft injury during ACR with CLAD or death remains undefined. Objectives: To investigate the association of ACR with the risk of CLAD or death and to further investigate if this risk depends on the degree of molecular allograft injury. Methods: This multicenter, prospective cohort study included 188 lung transplant recipients. Subjects underwent serial plasma collections for donor-derived cell-free DNA (dd-cfDNA) at prespecified time points and bronchoscopy. Multivariable Cox proportional-hazards analysis was conducted to analyze the association of ACR with subsequent CLAD or death as well as the association of dd-cfDNA during ACR with risk of CLAD or death. Additional outcomes analyses were performed with episodes of ACR categorized as "high risk" (dd-cfDNA ⩾ 1%) and "low risk" (dd-cfDNA < 1%). Measurements and Main Results: In multivariable analysis, ACR was associated with the composite outcome of CLAD or death (hazard ratio [HR], 2.07 [95% confidence interval (CI), 1.05-4.10]; P = 0.036). Elevated dd-cfDNA ⩾ 1% at ACR diagnosis was independently associated with increased risk of CLAD or death (HR, 3.32; 95% CI, 1.31-8.40; P = 0.012). Patients with high-risk ACR were at increased risk of CLAD or death (HR, 3.13; 95% CI, 1.41-6.93; P = 0.005), whereas patients with low-risk status ACR were not. Conclusions: Patients with ACR are at higher risk of CLAD or death, but this may depend on the degree of underlying allograft injury at the molecular level. Clinical trial registered with www.clinicaltrials.gov (NCT02423070).

Enhanced renal clearance impacts levetiracetam concentrations in patients with traumatic brain injury with and without augmented renal clearance

BackgroundThe purpose of this study was to examine the impact of ARC on levetiracetam concentrations during the first week following acute TBI. The hypothesis was levetiracetam concentrations are significantly lower in TBI patients with augmented renal clearance (ARC) compared to those with normal renal clearance.MethodsThis is a prospective cohort pharmacokinetic study of adults with moderate to severe TBI treated with levetiracetam during the first week after injury. Serial blood collections were performed daily for analysis of levetiracetam, cystatin C, and 12-hr creatinine clearance (CrCl) determinations. Patients were divided into two cohorts: with (CrCl ≥130 ml/min/1.73 m2) and without ARC.ResultsTwenty-two patients with moderate to severe TBI were included. The population consisted primarily of young male patients with severe TBI (mean age 40 years old, 68% male, median admission GCS 4). Each received levetiracetam 1000 mg IV every 12 h for the study period. ARC was present in 77.3% of patients, with significantly lower levetiracetam concentrations in ARC patients and below the conservative therapeutic range (< 6mcg/mL) for all study days. In patients without ARC, the serum concentrations were also below the expected range on all but two study days (Days 4 and 5). Four of the 22 (18.2%) patients exhibited seizure activity during the study period (two of these patients exhibited ARC). Cystatin C concentrations were significantly lower in patients with ARC, though the mean for all patients was within the typical normal range.ConclusionsARC has a high prevalence in patients with moderate to severe TBI. Levetiracetam concentrations after standard dosing were low in all TBI patients, but significantly lower in patients with ARC. This study highlights the need to consider personalized drug dosing in TBI patients irrespective of the presence of ARC.Clinical trial registrationThis study was registered at cliicaltrials.gov (NCT02437838) Registered on 08/05/2015, https://clinicaltrials.gov/ct2/show/NCT02437838.

A new approach for serial electron diffraction data collection.

This commentary describes a novel method for serial electron diffraction data collection in electron crystallography, utilizing a scanning transmission electron microscope to rapidly obtain patterns with low radiation dose. This approach, demonstrated with zeolite samples, has the potential to provide highly automated and rapid structures from nanocrystalline materials.

Characterizing the acute antibody response of monkeypox and MVA-BN vaccine following an Australian outbreak.

In response to the emergence of the monkeypox virus (MPXV) in Australia in May 2022, we developed and evaluated indirect immunofluorescence assays (IFA) for MPXV and Vaccinia virus (VACV) IgG and IgM antibodies using serum samples from patients with nucleic acid amplification test (NAAT)-confirmed mpox and uninfected unvaccinated controls. Additionally, 47 healthcare workers receiving two doses of the third-generation smallpox vaccine Modified Vaccinia Ankara-Bavarian Nordic (MVA-BN) undertook serial serum collection to describe the serological response to vaccination. MPXV antibodies were detected in 16/18 individuals with NAAT-confirmedmpox (sensitivity 0.89, specificity 1.00), and VACV antibodies were detected in 28/29 individuals who received two doses of MVA-BN vaccine (sensitivity 0.97, specificity 1.00). Detectable antibody in subjects historically vaccinated with early-generation vaccines against smallpox was found in 7/7 subjects, at a median of 48 years following vaccination. MPXVNAAT-positivepatients with serum samples collected within the first 14 days after rash onset had detectable IgG and IgM in 9/12 and 5/12 of patients, respectively, with maintenance of IgG and disappearance of IgM titersafter 60 days. Whilespecificity was high when testing unvaccinated and uninfected subjects, significant cross-reactivity between MPXV and VACV antibodies was observed.

Prospective healthcare-associated links in transmission of nontuberculous mycobacteria among people with cystic fibrosis (pHALT NTM) study: Rationale and study design.

Healthcare-associated acquisition and transmission of nontuberculous mycobacteria (NTM) among people with cystic fibrosis (pwCF) has been described, and remains a concern for both patients and providers. This report describes the design of a prospective observational study utilizing the standardized epidemiologic investigation toolkit for healthcare-associated links in transmission of NTM among pwCF. This is a parallel multi-site study of pwCF who have infections with respiratory NTM isolates and receive healthcare within a common CF Care Center. Participants have a history of one or more NTM positive airway cultures and have been identified as having NTM infections suggestive of a possible outbreak within a single Center, based on NTM isolate genomic analysis. Participants are enrolled in the study over a 3-year period. Primary endpoints are identification of shared healthcare-associated source(s) among pwCF in a Center, identification of healthcare environmental dust and water biofilm NTM isolates that are genetically highly-related to respiratory isolates, and identification of common home of residence watersheds among pwCF infected with clustered isolates. Secondary endpoints include characterization of healthcare-associated transmission and/or acquisition modes and settings as well as description of incidence and prevalence of healthcare-associated environmental NTM species/subspecies by geographical region. We hypothesize that genetically highly-related isolates of NTM among pwCF cared for at the same Center may arise from healthcare sources including patient-to-patient transmission and/or acquisition from health-care environmental dust and/or water biofilms. This study design utilizes a published, standardized, evidence-based epidemiologic toolkit to facilitate confidential, independent healthcare-associated NTM outbreak investigations within CF Care Centers. This study will facilitate real-time, rapid detection and mitigation of healthcare-associated NTM outbreaks to reduce NTM risk, inform infection prevention and control guidelines, and characterize the prevalence and origin of NTM outbreaks from healthcare-associated patient-to-patient transmission and/or environmental acquisition. This study will systematically characterize human disease causing NTM isolates from serial collection of healthcare environmental dust and water biofilms and define the most common healthcare environmental sources harboring NTM biofilms. ClinicalTrials.gov NCT05686837.

Abstract PR018: Feasibility of serial microbiome collection in a multi-site neoadjuvant pancreatic cancer clinical trial allowing for a diverse patient population

Abstract Background: Pancreatic cancer portends a very poor prognosis. It is the third leading cause of cancer deaths in the United States. African Americans have a 50- 90% increased incidence of pancreatic cancer in comparison to other racial groups. 20% of patients with pancreatic cancer present with early-stage disease and will have a median overall survival of 2.5 years if able to undergo surgical resection. There is a desperate need to discover novel ways of treating pancreatic cancer. Understanding of the microbiome may be informative to overcoming chemotherapy resistance, toxicity and improving outcomes and disparities. The aim of this study is to (1) identify changes in the intestinal microbiota during neoadjuvant treatment of pancreatic ductal adenocarcinoma that correlate with treatment response and (2) identify changes in the intestinal microbiota during neoadjuvant treatment of pancreatic ductal adenocarcinoma that correlate with treatment toxicity in a diverse patient population. Methods: Patients with resectable or borderline resectable pancreatic cancer who are enrolled in a multi-site clinical trial (NCT03483038) receiving neoadjuvant NEO-Nali-IRI (oxaliplatin, liposomal irinotecan and 5- FU) followed by surgical resection were asked to provide serial microbiome collections of saliva, urine, blood, and stool with a dietary log during the following four (or five) time points: at baseline, mid-therapy (after C4), at completion of chemotherapy (after C8), after radiation if applicable, and at 4-6 weeks post-operative. Participants were given stool collection kits during clinic visits for home sample acquisition. All stool samples were mailed to the University of Florida Health Cancer Center Microbiome Biorepository. Saliva, urine and blood were collected during clinic visits. Demographics, stage, medical history and concurrent medications including antimicrobics and probiotics, chemotherapy dose-reductions, delays, discontinuations, adverse events, safety adverse events, attribution of causality, and hospitalizations were obtained. Outcomes include imaging response using RECIST criteria, CA 19-9 trend, development of metastatic disease, and the ability to proceed to definitive surgical resection. DNA will be extracted from stool and analyzed using 16S rRNA sequencing methods to determine signatures and biodiversity. Descriptive statistics and correlative analysis will be reported. Continuous and categorical variables will be compared with Student’s t-test and Chi-squared test. Multiple groups will be compared by ANOVA. Kaplan-Meier logistic regression will be performed to compare outcomes between the different microbiota groupings. Conclusion: Serial microbiome collection from a multi-site clinical trial is feasible during neoadjuvant chemotherapy for pancreatic cancer and can increase the diversity of the sample population. Citation Format: Sherise C. Rogers, Ibrahim Nassour, Ryan Thomas, Ilyas Sahin, Brian Ramnaraign, Karen Russell, Steven Hughes, Kathryn Hitchcock, Omar Kayaleh, Anita Turk, Ji-Hyun Lee, David DeRemer, Carmen Allegra, Thomas George. Feasibility of serial microbiome collection in a multi-site neoadjuvant pancreatic cancer clinical trial allowing for a diverse patient population [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr PR018.

Vascular, Imaging and Cognition Association of China (VICA) Study: Rationale and Design

AbstractBackgroundChina has the highest number of patients with dementia in the world, and about 15‐30% of these cases are caused by vascular disease. Currently, the pathophysiological mechanism that cause vascular cognitive impairment (VCI) remain insufficiently understood. Therefore, early diagnosis and prognostic indicators of VCI are critically important for managing its onset and development To this end, several Chinese institutions have joined forces to establish “Vascular, Imaging and Cognition Association of China” (VICA) (Figure 1). The objective of VICA is to identify new targets for intervention and prompting intelligent graded and stratified treatment.MethodVICA is the first large‐scale multi‐center VCI consortium in China. It consists of seven advanced centers and over two hundred primary hospitals across the country. Included VICA are three different populations at risk for VCI: (a) high vascular risk factors for dementia, (b) cerebral small vessel disease, and (c) acute ischemic stroke or transient ischemic attacks (TIA).ResultAll individuals will be enrolled and monitored for at least 1 year using serial neuropsychological assessment, neuroimaging and collection of multi‐omics data from biospecimens (e.g., blood, urine, feces). In addition, a small sample of patients will undergo PET scans to determine whether neurodegenerative pathology (such as amyloid or tau accumulation) contributes to the course of vascular dementia. The enrollment procedure of patients with stroke or TIA is shown in Figure 2.ConclusionThe overall scientific aim of VICA is to guide early clinical diagnosis and early intervention of VCI based on the integration of multiple vascular risk factors, neuroimaging features and biomarkers. VICA represents a unique scientific resource that will help better understand the dynamics of cognitive change in different populations and identify new potential therapeutic targets for this condition.

Abstract C025: Colli-Pee™ UAS™ 40mL: A first-void urine collection device with potential to improve access to cancer screening when paired with an appropriate clinical assay

Abstract In recent years, liquid biopsy has become increasingly important for cancer screening, detection and monitoring as cancer-specific biomarkers can be captured non-invasively in biological fluids. First-void urine (FVU) offers distinct advantages, such as ease of collection and large available volumes. Colli-Pee™ UAS™ 40 mL is a FVU self-collection device pre-filled with UAS, a non-lytic liquid preservative. Usability questionnaires from 98 healthy and diseased users showed that Colli-Pee™ was clear (90%) and comfortable (68%) to use, underscoring its potential to improve access to cancer screening for underserved communities. Here, we evaluated the performance of the device for FVU collection and urinary analyte preservation. FVU samples were collected from healthy donors with the Colli-Pee™ UAS 40 mL device to evaluate analyte preservation (n=14). Samples were stored at room temperature (RT), with aliquots processed at baseline and 8 days, or after freeze/thaw cycling (-20°C/-4°F to 40°C/104°F) to simulate ambient temperature transport. Cell-free DNA (cfDNA, Qiagen Circulating Nucleic Acid Kit) and extracellular vesicle RNA (EV RNA, Qiagen exoRNeasy Maxi Kit) were extracted from sample supernatants and evaluated for analyte preservation via qPCR assays targeting the β-globin gene (cfDNA) or the GAPDH mRNA (EV RNA). Cellular pellets were obtained at each time point by centrifugation and genomic DNA was extracted (Qiagen PowerFecal Pro DNA Kit) to evaluate both host cell integrity and microbial growth. Both were inferred based on the amount of change in qPCR signal for a human DNA target (TS143) and a non-specific bacterial target (16S). Data are presented as mean ± SD. Analysis of RT-stored samples confirmed that both urinary cfDNA and EV RNA were well preserved (ΔCt: 0.01 ± 1.04 and 0.58 ± 1.15, respectively). Similarly, qPCR of host and microbial DNA recovered from the cellular pellet showed no change after storage (ΔCt: 0.00 ± 0.50 and 0.06 ± 0.51, respectively), demonstrating that host cells remained intact and that UAS™ preservative prevented microbial growth. Under simulated ambient temperature transportation conditions, cfDNA (ΔCt: 0.57 ± 1.58), EV RNA (1.05 ± 0.94), and host cell integrity (0.63 ± 0.73) were successfully maintained. Urinary cfDNA, EV RNA, and host cell integrity from FVU samples were effectively preserved up to 7 days post-collection at RT, and under simulated ambient temperature transportation. By enabling standardized volumetric FVU collection and analyte preservation, the Colli-Pee™ UAS™ 40 mL device demonstrates its viability as a non-invasive FVU self-sampling device for oncology applications, simplifying serial and at-home collection. In addition to the advantages of FVU, Colli-Pee™ collection devices can improve access to testing by allowing for samples to be collected at home and transported at ambient temperature, which can improve the chances of early detection and outcomes for diverse patients in at-risk populations. Citation Format: Kyle MacDonald, Melissa Richer, Thanh Dang, Alejandra Rioscortes, Cameron Wood, Nette Meers, Savannah Colameco, Koen Beyers, Graham Morse, Tara Crawford Parks. Colli-Pee™ UAS™ 40mL: A first-void urine collection device with potential to improve access to cancer screening when paired with an appropriate clinical assay [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C025.

Dysfunctional Stromal Pathways Mark Early Changes within the Bone Marrow Microenvironment in JAK2 V617F-Driven Model of Myeloproliferative Neoplasm

Background: JAK2V617F (JAK2VF) is the primary driver mutation in classical myeloproliferative neoplasms (MPN). JAK2VF is present decades prior to diagnosis (Mitchell et.al. Nature 2022), and the JAK2VF+ clone may remodel the BM microenvironment further supporting its expansion (Curto-Garcia, et. al. Haematologica 2020). Understanding this relationship, particularly during early disease development may aid identification of new therapeutic targets to stop or revert these early changes. Human BM procured before MPN diagnosis is rarely available and most murine MPN models use myeloablative conditioning (eg., irradiation), permanently altering the BM stroma (Costa &amp; Reagan Frontiers 2019), diminishing the clinical relevance of resultant findings. We developed an innovative translatable model of MPN with sustainable low-level disease engraftment and no requirement for recipient conditioning. By preserving the normal microenvironment, our model enabled detection of early changes within the BM stroma induced by the JAK2VF+ clone. Methods: Whole BM transplantation (BMT) was performed via a single tail vein injection into unconditioned C57BL/6 Ptprc a (JAX#002014, CD45.1+) recipient mice. Donor animals consisted of either wildtype ( wt) C57BL/6 (JAX#000664) mice or our PolyI:C inducible MPN-like model created by crossing a floxed-JAK2VF (JAX#031658) with Mx1-Cre (JAX#003556) animals (JAK2VF Parental). For all BMTs, donors and recipients were age- (10wks) and sex-matched. BMT cell concentrations ranged from 1e6 to 15e6per injection, n≥10 mice, both sexes were used per group. A cohort of non-BMT age- and sex-matched recipients were maintained as naïve controls. Recipient serial submandibular blood collections were used to examine complete blood cell counts and donor chimerism. Bulk RNA-seq transcriptomic analysis was performed on digested and lineage-negative sorted whole BM from matched naïve and 5e6 BMT recipients (n=7 and 6, respectively) to delineate stromal alterations driven by JAK2VF+ MPN clone. Qualitative x-ray analysis was performed on femurs. Results: 15e6 wt donor CD45.2 BM cells can be transplanted sustainably into unconditioned CD45.1 recipients with ~3.0% donor chimerism, maintained up to 200-days post-BMT (Fig A). Follow up secondary BMT into conditioned recipients using as few as 25e4CD45.2+ cells sorted from primary BMT recipients displayed 83% average engraftment by 150-day post BMT, confirming functional wt donor CD45.2 HSC. This successful strategy was used to perform BMT using JAK2VF parental (1e6, 5e6, and 15e6) cells. Detected chimerism followed a dose response, not exceeding an average of 10% up to &amp;gt;225 days. Surprisingly, with the engraftment at &amp;lt;10%, we observed MPN-like phenotype (elevated hemoglobin and monocytosis) except for no thrombocytosis (Fig A). To identify alterations in the BM niche, stromal cells were isolated from BMT recipients and subjected to RNA-seq. Enrichment of stromal cells was validated by confirming that 23% of the top 100 highest read coverage transcripts were encoding stromal genes (i.e. Sparc, Dcn, Col2a1, and Cxcl12). Gene set enrichment analysis (GSEA) of stromal fraction showed upregulation of osteoclastic differentiation and bone remodeling. GSEA against hallmark gene sets indicated upregulation of mesenchymal phenotype and inflammatory response suggesting an underlying effect on the stroma driven by the presence of the JAK2VF clone. Qualitative x-ray of femurs indicated loss of bone density, suggesting disease-driven effect on the stroma (Fig B). Discussion: The development of our translatable model of MPN, where a small subset of cells drives the disease within unconditioned recipients represents an important milestone towards studying early disease pathology in MPNs. Using wt BMT, we demonstrated the successful establishment of long-term, low-level chimerism in unconditioned recipients. We found the clone-dependent impact on the stroma without myeloablative conditioning. Bulk transcriptomics and femur x-rays indicate aberrant bone morphology in BMT mice, despite the low level of engraftment (&amp;lt;5%) in the recipient system. Our data suggests that even low VAF of JAK2VF may support pathologic remodeling of the BM environment, warranting further exploration of patient BM niche early and late in the MPN course.

Field outcomes of laparoscopic ovum pick-up combined with in vitro embryo production in sheep: Effects of long-acting recombinant ovine FSH pre-stimulation, collection frequency, and donor breed

Laparoscopic ovum pick-up (LOPU) combined with in vitro embryo production (IVEP) is a technology platform that improves the utilization rate of the elite ewe's ovarian oocytes and increases the number of obtained offspring. This study aimed to evaluate the effects of FSH pre-stimulation, serial oocyte collection, and breed on LOPU-IVEP under field conditions. Donors were randomly assigned to five groups (group A: decreasing doses of pituitary FSH (p-FSH); group B: constant doses of p-FSH; group C: two doses of long-acting recombinant ovine FSH (ro-FSH); group D: single administration of a long-acting ro-FSH in; group E: no FSH stimulation). Oocyte yield following LOPU (average recovered oocytes: 20.9 ± 0.5; average viable oocytes: 17.2 ± 0.4) and oocyte developmental competence (average blastocysts: 7.0 ± 0.2) in group C were significantly better than these of group D and group E, and similar to these of groups A and B. Meanwhile, there were no differences in oocyte yield and developmental capacity using repeated LOPU session at 1-, 2-, and 3-month intervals (p > 0.05). Finally, we compared LOPU-IVEP outcomes among five sheep breeds. The results indicated that East Friesian × Chinese Mongolian crossbred sheep and purebred East Friesian sheep had the more recovered oocytes and viable oocytes compared with the Suffolk, Dorper, and Texel breeds, and average number of blastocysts in East Friesian × Chinese Mongolian sheep group was also highest among the groups (8.1 ±0.3, p < 0.05). In summary, the results of this study indicate long-acting ro-FSH pre-stimulation combined with 12 times LOPU sessions over one year maximizes embryo production of elite donor ewes under field conditions.

Serum Titanium Levels Remain Elevated But Urine Titanium is Undetectable in Children With Early Onset Scoliosis Undergoing Growth-Friendly Surgical Treatment: A Prospective Study

Background: Elevated serum titanium levels have been found in patients with early onset scoliosis (EOS) treated with traditional growing rods (TGR), magnetically controlled growing rods (MCGR), and vertical expandable prosthetic titanium rib (VEPTR). No studies have investigated whether serum titanium remains persistently elevated and if titanium is excreted. Our purpose was to compare serum titanium levels in patients with EOS with growth-friendly instrumentation to age-matched controls and evaluate urine titanium and serial serum titanium levels in patients with EOS. Methods: This was a prospective case-control study. Patients with EOS with TGR, MCGR, or VEPTR underwent urine titanium and serial serum titanium collection at a minimum 6-month interval. Control patients did not have a history of metal implant insertion and underwent serum titanium collection before fracture fixation. Results: Twenty patients with EOS (6 TGR, 8 MCGR, and 6 VEPTR) and 12 controls were analyzed. The control group had no detectable serum titanium (0 ng/mL), whereas the patients with EOS had a median serum titanium of 4.0 ng/mL (P &lt; 0.001). Analysis of variance showed significantly higher median serum titanium levels in the MCGR and VEPTR groups than the TGR group at time point 1 (5.5 vs 6.0 vs 2.0 ng/mL, P = 0.01) and time point 2 (6.5 vs 7.5 vs 2.0 ng/mL, P &lt; 0.001). Binary comparisons showed a significant difference in serum titanium level between TGR and MCGR (time point 1: P = 0.026, time point 2: P = 0.011) and TGR and VEPTR (time point 1: P = 0.035, time point 2: P = 0.003). However, there was no difference between MCGR and VEPTR (time point 1: P = 0.399, time point 2: P = 0.492) even though the VEPTR group had a longer duration of follow-up (P = 0.001) and a greater number of lengthenings per patient at the first serum collection (P = 0.016). No patients with EOS had detectable urine titanium. Conclusions: Patients with EOS treated with titanium alloy growth-friendly instrumentation had elevated serum titanium levels compared with age-matched controls that persisted over time with no evidence of renal excretion. Additional studies are necessary to assess for local and systemic accumulation of titanium and the significance of long-term exposure to titanium in growing children. Level of Evidence: Level III, therapeutic.

EXTH-56. CSF PENETRATION COMPARISON OF TWO ORAL FORMULATIONS OF APL-101 IN A PRE-CLINICAL NON-HUMAN PRIMATE MODEL

Abstract Vebreltinib (APL-101) is a potent c-Met inhibitor which has shown efficacy in murine pre-clinical models and is under clinical trial evaluation for several cancer types, including adult high-grade glioma. The capsule formulation (APL-101-Cap) was reformulated as a pill (APL-101-Pill) with the expectation of increasing bioavailability. We determined and compared the plasma pharmacokinetics (PK) and cerebrospinal fluid (CSF) penetration of APL-101-Cap and APL-101-Pill in a non-human primate (NHP) model. METHODS: Seven animals, previously developed as serial CSF access models, were utilized across all studies. One NHP was studied with both formulations. Both formulations were administered orally as a single drug/single dose in serial studies, followed by serial plasma and CSF sample collections from 0-96 hours. Dosages: APL-101-Cap 3.25 mg/kg (n = 3) or 6.5 mg/kg (n = 4) and APL-101-Pill 5 mg/kg (n = 4). The resulting plasma concentrations from each formulation were quantified by LC-MS/MS. PK parameters were calculated via noncompartmental methods, and results were analyzed to assess CNS penetration (AUCCSF:AUCPlasma) and compare formulations. RESULTS: Mean (range) for plasma and CSF PK exposure [AUCLAST (hr*ng/mL)] reported. APL-101-Cap plasma exposure, regardless of dose, was more variable than APL-101-Pill. APL-101-Pill had lower exposure in both plasma (AUC 1893.0 (841-2865)] and CSF [AUC 27.7 (5.0-65.0)] than APL-101-Cap regardless of dose [(3.25 mg/kg): plasma AUC 9621.0 (1331-23132) and CSF AUC 688.0 (24-1352); 6.5 mg/kg: plasma AUC 6208.0 (1424-19800) and CSF AUC 606.0 (34-1179)]. CNS penetrance was higher with APL-101-Cap (3.2% [0.5-5.8%] at 3.25mg/kg; 4.2% (2.4-6%) at 6.5 mg/kg), but also more variable when compared to APL-101-Pill (1.2% [0.6-2.3%]). For APL-101-Cap, one animal at 3.25 mg/kg and two animals at 6.25 mg/kg had undetectable CSF concentrations. CONCLUSION: In this NHP model, single-dose oral administration of APL-101-Pill resulted in lower, but less variable plasma and CSF exposure compared to APL-101-Cap. APL-101-Cap CSF exposure was unaffected by dose escalation.

CTNI-07. PHASE I STUDY OF PAXALISIB AND RADIOTHERAPY FOR CNS DISEASE HARBORING PI3K PATHWAY MUTATIONS: PILOT ANALYSIS OF CIRCULATING TUMOR DNA FOR PATIENT ELIGIBILITY CONFIRMATION AND POST-TREATMENT RESPONSE

Abstract INTRODUCTION Radiotherapy (RT) is effective for brain metastases (BM) or leptomeningeal disease (LMD), but disease control is poor for tumors harboring PI3K pathway alterations (PI3Kalt). We hypothesized that combining RT with paxalisib, a CNS-penetrant PI3K/mTOR inhibitor, offers therapeutic synergy. The role of plasma circulating tumor DNA (ctDNA) to confirm study eligibility and response monitoring is unclear. METHODS This is a multi-institutional, phase I trial of concurrent paxalisib and whole brain RT (30 Gy in 10 fractions, NCT04192981) for CNS disease. To establish eligibility, patients had documented PI3Kalt performed using institutional tumor targeted sequencing (MSK-IMPACT) of primary or metastatic tumors (eligibility sequencing). Plasma ctDNA was collected at baseline and 3-, 6- and 9-months post-treatment and sequenced using institutional plasma ctDNA targeted sequencing platform (MSK-ACCESS). Our objectives were to a) determine PI3Kalt concordance between eligibility sequencing and baseline ctDNA and b) analyze quantitative post-treatment plasma ctDNA dynamics. RESULTS Between 3/2020-5/2023, 15 patients were enrolled to receive daily paxalisib (45mg, n = 11 or 60mg, n = 4) and WBRT. Twelve were treated for BM and 3 for LMD. Eligibility sequencing was determined a median of 8.1months (range: 2.4-38.6) pre-study enrollment and the most common PI3Kalt were PIK3CA (n = 7), INPP4B (n = 2), INPPL1 (n = 1) and PIK3CG (n = 1), and PTEN copy number alterations (homdel n = 2). Baseline ctDNA was sequenced from 13/15 patients (18 samples: baseline = 13, 3-months = 4 and 6-months = 1). We confirmed 100% of PIK3CA mutations using baseline ctDNA. One breast cancer BM patient demonstrated significant variant allele frequency reduction from 50% (baseline) to 0.1% (3-months post-treatment) for two tracked PIK3CA mutations (p.E545K and p.A1066V), which correlated with a partial response per RANO-BM. CONCLUSIONS For patients on genomically-driven targeted therapy and RT combination trials, alterations detected using plasma ctDNA, especially PIK3CA, may be used to rapidly assess eligibility. Serial ctDNA collection may serve as a potential response biomarker with further validation.

CTIM-40. EARLY RADIOGRAPHIC RESPONSE AND ENGRAFTMENT OF PHASE I FIRST-IN-HUMAN BICISTRONIC CAR T CELLS CORRELATE WITH REAL-TIME AUTOLOGOUS GBM ORGANOID CYTOLYSIS

Abstract Glioblastoma (GBM) is a highly treatment-resistant cancer, in part due to prominent inter- and intratumoral heterogeneity. To date, monovalent CAR T-cell therapies have failed to overcome this heterogeneity. Bivalent CAR-T cells may overcome this challenge by expanding the population of target-positive tumor cells. NCT05168423 is first-in-human, phase 1 dose escalation study to assess safety and feasibility of intrathecal delivery of autologous bicistronic CAR-T cells, targeting both EGFR and IL13Rα2, in patients with EGFR-amplified recurrent GBM. For each patient enrolled in this study, high-fidelity glioblastoma organoids (GBOs) are derived from tumor tissue and utilized to test the autologous CAR-T product in real-time. GBOs are co-cultured with autologous CAR-T cells and individually monitored using modified impedance spectroscopy assays over six days for quantification of real-time tumor cytolysis. CAR-T efficacy in GBOs was correlated to matched clinical trial patient outcomes, including CAR-T engraftment. To date, three patients have been treated at the initial dose level (10,000,000 CART-EGFR-IL13Rα2 cells). All three patients demonstrated tumor shrinkage on MRI obtained within 24-48hrs post-infusion. MRI changes were accompanied by significant tumor-inflammation associated neurotoxicity that resolved with steroids and IL1R blockade. Serial CSF collection for qPCR following CAR-T infusion revealed average peaks of &amp;gt;100,000 copies of CAR-T cells/μg of gDNA, comparable to CART19-treated patients. Cytotoxicity values in GBOs strongly correlated with peak CSF engraftment levels of CAR-T cells in all three patients (r = 0.998, p = 0.037). In summary, we present striking radiographic and biological responses from the first cohort of recurrent GBM patients treated with a bivalent, intrathecally delivered CAR T cell product simultaneously targeting EGFR and IL13Rα2. Potent anti-tumor activity of CART-EGFR-IL13Rα2 cells was also observed by treating patient-derived GBOs with the patients’ corresponding autologous CAR T-cell products, providing a parallel, ex vivo system for real-time prediction of CAR T-cell engraftment.

Serials Analysis Directions Part 2: Finding Gems, Pulling Weeds, and Reclaiming Space: Case Studies from GreenGlass for Serials

The presenters of the session, "Serials Analysis Directions Part 2: Finding Gems, Pulling Weeds, and Reclaiming Space: Case Studies from GreenGlass for Serials" provided attendees with valuable perspectives on using GreenGlass for Serials to conduct serials collection review projects effectively, efficiently, and with more confidence. The first two presenters, Katy Gabrio and Ann Roll, are librarians who have used GreenGlass for Serials. They shared their reasons for reviewing serials collections, their libraries' goals for the projects, and the results. The third speaker, Andy Breeding, a senior product manager at OCLC, spoke about the history of GreenGlass and GreenGlass for Serials, as well as the time savings gained from using GreenGlass for Serials, and trends he noticed in serials holdings of libraries through the years.

Serials Analysis Directions: The importance of Standard Numbers in Collaborative Collection Development for Serials

The Research Collections and Preservation Consortium (ReCAP) partners contracted the Center for Research Libraries (CRL) to do a volume level analysis of their combined print serial collections. This paper focuses on the decision to use the OCLC number as the primary match point and the starting point for the collection analysis, challenges encountered due to inconsistent OCLC number use across collections, the way those challenges informed the overall project, and the method CRL created to find OCLC numbers for records without them. Final results of the volume-level analysis, ongoing work to integrate Harvard’s serials into the analysis, and the effects of ReCAP partners’ collaborative collection development initiatives are included.

Duration of Enterovirus D68 RNA Shedding in the Upper Respiratory Tract and Transmission among Household Contacts, Colorado, USA.

Enterovirus D68 (EV-D68) causes cyclical outbreaks of respiratory disease and acute flaccid myelitis. EV-D68 is primarily transmitted through the respiratory route, but the duration of shedding in the respiratory tract is unknown. We prospectively enrolled 9 hospitalized children with EV-D68 respiratory infection and 16 household contacts to determine EV-D68 RNA shedding dynamics in the upper respiratory tract through serial midturbinate specimen collections and daily symptom diaries. Five (31.3%) household contacts, including 3 adults, were EV-D68-positive. The median duration of EV-D68 RNA shedding in the upper respiratory tract was 12 (range 7-15) days from symptom onset. The most common symptoms were nasal congestion (100%), cough (92.9%), difficulty breathing (78.6%), and wheezing (57.1%). The median illness duration was 20 (range 11-24) days. Understanding the duration of RNA shedding can inform the expected rate and timing of EV-D68 detection in associated acute flaccid myelitis cases and help guide public health measures.

Serum Anti-Spike Antibodies Are Not Affected by Immunosuppressants in SARS-CoV-2 Vaccinations Given to Brazilian Patients with Inflammatory Bowel Disease.

This study aimed to evaluate humoral responses after vaccination against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) of patients with inflammatory bowel disease (IBD). Patients with IBD enrolled in a tertiary outpatient unit were followed up between September 2021 and September 2022 via serial blood collection. Immunoglobulin G antibody titers against SARS-CoV-2 were measured before administration and 1 and 6 months after the administration of two doses of different vaccination regimens. The results were compared with those of a healthy control group obtained during the same period. The mean pre-vaccination antibody titers were 452.0 and 93.3 AU/mL in the IBD (n = 42) and control (n = 89) groups, respectively. After two doses of the vaccine, the titers significantly increased in both groups (IBD, 8568.0 AU/mL; control, 7471.0 AU/mL; p < 0.001). One month after the second dose, no significant differences were observed between the two groups (p = 0.955). Significant differences between vaccination schemes in the IBD group were observed, with higher titers in those who received Pfizer, younger patients (p < 0.005), and those with a previous coronavirus disease 2019 (COVID-19) infection (p < 0.012). The use of immunosuppressants and immunobiologicals did not affect the overall humoral response to COVID-19 vaccine in patients with IBD, but specific vaccine regimens, age, and previous coronavirus infection significantly did. This study reinforces the positive impact of booster doses and the safety of SARS-CoV-2 vaccination.