T1 Samples Research Articles

Real-time automated quality control for quantitative MRI.

This work presents an automated quality control (QC) system within quantitative MRI (qMRI) workflows. By leveraging the ISMRM/NIST quantitative MRI system phantom, we establish an open-source pipeline for rapid, repeatable, and accurate validation and stability tracking of sequence quantification performance across diverse clinical settings. A microservice-based QC system for automated vial segmentation from quantitative maps was developed and tested across various MRF acquisition and protocol designs, with reports generated and returned to the scanner in real time. The system demonstrated consistent and repeatable value segmentation and reporting, successfully extracted all 252 T1 and T2 vial samples tested. Values extracted from the same sequence were found to be repeatable with 0.09% ± 1.23% and -0.26% ± 2.68% intersession error, respectively. By providing real-time quantification performance assessment, this easily deployable automated QC approach streamlines sequence validation and long-term performance monitoring, vital for the broader acceptance of qMRI as a standard component of clinical protocols.

Effect of Deposit Scale on Mechanical Properties of In-Situ Alloyed CrCoNi Medium Entropy Alloys Formed by Directed Energy Deposition

Directed energy deposition (DED), as an additive manufacturing technology, has shown unique advantages in multi-material additive manufacturing and remanufacturing. In this study, two types in-situ alloyed CrCoNi medium entropy alloys that have thin-walled structures with different thicknesses (T1 and T2) were manufactured by the DED process, and the mechanisms of differences in relative density, microstructure, and mechanical properties at different heights were systematically analyzed. In terms of microstructure, the T1 and T2 samples along the building direction exhibit significant differences in crystallographic orientation, grain size, and dislocation density, which are related to the local temperature gradient differences caused by the scanning path and heat accumulation. In terms of mechanical properties at different heights of the two types of thin-walled structures, the yield strength is higher but the elongation is lower at the bottom position of sample, while the yield strength is lower but the elongation is higher at the middle and top positions. The differences of mechanical properties at different heights of the T1 and T2 samples are related to the microstructure and relative density. This finding provides new insights for the design and performance analysis of complex thin-walled structures formed by additive manufacturing.

Physico-chemical changes in developed probiotic chicken meat spread fermented with Lactobacillus acidophilus and malted millet flour

This study was envisaged to develop a probiotic chicken meat spread fermented with Lactobacillus acidophilus (Lactic acid bacteria-LAB) and malted sorghum flour as a substrate and evaluation of its quality. Chicken meat spread was prepared by incorporating malted sorghum flour at 0, 2, 4 and 6 % (w/w) levels as a substrate in the product's formulation, along with Lactobacillus acidophilus at a concentration of 1 million cfu/g of the product. The product prepared with different formulations i.e. C (meat spread only), C1 (meat spread with 0 % malted sorghum flour + LAB), T1 (meat spread with 2 % malted sorghum flour + LAB), T2 (meat spread with 4 % malted sorghum flour + LAB) and T3 (meat spread with 6 % malted sorghum flour + LAB) were assessed for fresh product study and the storage stability at refrigeration temperature for 16 days when the physico-chemical parameters were analyzed. The fresh product study revealed significantly (p ≤ 0.05) higher moisture content in the T3 group. However, lower protein, ether extract, and ash content in T2 and T3 samples compared with the control, whereas a higher emulsion stability and cooking yield in the T2 group were observed when compared with the control. Free fatty acid, thiobarbituric acid, and titratable acidity values significantly (p ≤ 0.05) increased for the fermented samples in comparison to the control, and the same trend continued during the entire storage period. This study showcases the potential of using malted sorghum flour and Lactobacillus acidophilus to enhance the functional properties of chicken meat spreads, making them more nutritious and appealing to health-conscious consumers. Future research should focus on optimizing ingredient proportions and exploring alternative substrates to further improve these functional meat products.

Integrating omics techniques and culture-independent systems may improve the detection of persistent candidemia: data from an observational study

IntroductionBlood cultures have low sensitivity for candidemia. Sensitivity can be improved by the culture-independent system T2 Magnetic Resonance (T2). SeptiCyte RAPID is a host response assay quantifying the risk of infection-related inflammation through a scoring system (SeptiScore). We investigate the performance of SeptiScore in detecting persistent candidemia as defined by conventional cultures and T2.MethodsThis is a prospective multicentre observational study on patients with candidemia. Blood cultures and blood samples for assessment by T2 and SeptiCyte were collected for 4 consecutive days after the index culture. The performance of SeptiScore was explored to predict persistent candidemia as defined by (1) positive follow-up blood culture (2) either positive follow-up blood culture or T2 sample.Results10 patients were enrolled including 34 blood collections assessed with the 3 methods. Overall, 4/34 (12%) follow-up blood cultures and 6/34 (18%) T2 samples were positive. A mixed model showed significantly higher SeptiScores associated with persistent candidemia when this was defined as either a positive follow-up blood culture or T2 sample (0.82, 95%CI 0.06 to 1.58) but not when this was defined as a positive follow-up blood culture only (-0.57, 95%CI -1.28 to 0.14). ROC curve for detection of persistent candidemia by SeptiScore at day 1 follow-up showed an AUC of 0.85 (95%CI 0.52-1.00) when candidemia was defined by positive follow-up blood culture, and an AUC of 1.00 (95%CI 1.00–1.00) when candidemia was defined according to both methods.ConclusionIntegrating transcriptome profiling with culture-independent systems and conventional cultures may increase our ability to diagnose persistent candidemia.

Circulating interleukin-8 and osteopontin are promising biomarkers of clinical outcomes in advanced melanoma patients treated with targeted therapy

BackgroundCirculating cytokines can represent non-invasive biomarkers to improve prediction of clinical outcomes of cancer patients. Here, plasma levels of IL-8, CCL4, osteopontin, LIF and BDNF were determined at baseline (T0), after 2 months of therapy (T2) and, when feasible, at progression (TP), in 70 melanoma patients treated with BRAF and MEK inhibitors. The association of baseline cytokine levels with clinical response, progression-free survival (PFS) and overall survival (OS) was evaluated.MethodsCytokine concentrations were measured using the xMAP technology. Their ability to discriminate between responding (Rs) and non-responding (NRs) patients was assessed by Receiver Operating Characteristics analysis. PFS and OS were estimated with the Kaplan–Meier method. The Cox proportional hazard model was used in the univariate and multivariate analyses to estimate crude and adjusted hazard ratios with 95% confidence intervals.ResultsCCL4 and LIF were undetectable in the majority of samples. The median osteopontin concentration at T0 and T2 was significantly higher in NRs than in Rs. The median T0 and T2 values of IL-8 were also higher in NRs than in Rs, although the statistical significance was not reached. No differences were detected for BDNF. In 39 Rs with matched T0, T2, and TP samples, osteopontin and IL-8 significantly decreased from T0 to T2 and rose again at TP, while BDNF levels remained unchanged. In NRs, none of the cytokines showed a significant decrease at T2. Only osteopontin demonstrated a good ability to discriminate between Rs and NRs. A high IL-8 T0 level was associated with significantly shorter PFS and OS and higher risk of progression and mortality, and remained an independent negative prognostic factor for OS in multivariate analysis. An elevated osteopontin T0 concentration was also significantly associated with worse OS and increased risk of death. Patients with high IL-8 and high osteopontin showed the lowest PFS and OS, and in multivariate analysis this cytokine combination remained independently associated with a three- to six-fold increased risk of mortality.ConclusionCirculating IL-8 and osteopontin appear useful biomarkers to refine prognosis evaluation of patients undergoing targeted therapy, and deserve attention as potential targets to improve its clinical efficacy.

Functional analysis of root-preferential oil palm metallothionein promoter in tobacco.

Root-specific or preferential promoters are essential to genetically modify plants with beneficial root traits. We have characterised the promoter from an oil palm metallothionein gene (EgMT) and performed a serial 5' deletion analysis to identify the region(s) essential for transgenes expression in roots. Stable functional characterisation of tobacco transgenic lines using the T1 generation showed that a deletion construct, designated as RSP-2D (1107bp), directed strong GUS expression at all stages of root development, particularly in mature roots. Other constructs, RSP-2A (2481bp) and RSP-2C (1639bp), drove GUS expression in roots with an intensity lower than RSP-2D. The promoter activity was also detectable in seed pods and immature seeds, albeit at lower levels than CaMV35S. The promoter activity may also be induced by wounding as intact GUS staining was observed at the flower- and leaf-cutting sites of T1 samples carrying either RSP-2C or RSP-2D constructs. The promoter sequence contains cis-acting elements that may act as negative regulators and be responsible for root specificity. The results further indicated that the 5' UTR and ATATT sequences are essential for strong promoter activity. This study highlights the potential of RSP-2D promoter as a tool for modifying root traits through genetic engineering.

Ultrashort echo time magnetic resonance elastography for quantification of the mechanical properties of short T2 tissues via optimal control-based radiofrequency pulses.

The aim of the current study is to demonstrate the feasibility of radiofrequency (RF) pulses generated via an optimal control (OC) algorithm to perform magnetic resonance elastography (MRE) and quantify the mechanical properties of materials with very short transverse relaxation times (T2 < 5 ms) for the first time. OC theory applied to MRE provides RF pulses that bring isochromats from the equilibrium state to a fixed target state, which corresponds to the phase pattern of a conventional MRE acquisition. Such RF pulses applied with a constant gradient allow to simultaneously perform slice selection and motion encoding in the slice direction. Unlike conventional MRE, no additional motion-encoding gradients (MEGs) are needed, enabling shorter echo times. OC pulses were implemented both in turbo spin echo (OC rapid acquisition with refocused echoes [RARE]) and ultrashort echo time (OC UTE) sequences to compare their motion-encoding efficiency with the conventional MEG encoding (classical MEG MRE). MRE experiments were carried out on agar phantoms with very short T2 values and on an ex vivo bovine tendon. Magnitude images, wave field images, phase-to-noise ratio (PNR), and shear storage modulus maps were compared between OC RARE, OC UTE, and classical MEG MRE in samples with different T2 values. Shear storage modulus values of the agar phantoms were in agreement with values found in the literature, and that of the bovine tendon was corroborated with rheometry measurements. Only the OC sequences could encode motion in very short T2 samples, and only OC UTE sequences yielded magnitude images enabling proper visualization of short T2 samples and tissues. The OC UTE sequence produced the best PNRs, demonstrating its ability to perform anatomical and mechanical characterization. Its success warrants in vivo confirmation in further studies.

Development of Ready-to-Eat Breakfast Flakes with Green Banana (&lt;em&gt;Musa Acuminata&lt;/em&gt;) and Jackfruit Seeds (&lt;em&gt;Artocarpus Heterophyllus&lt;/em&gt;) Flour: A Comprehensive Analysis of Nutritional and Functional Properties

Green banana flour (GBF) and jackfruit seed flour (JSF) present sustainable solutions for reducing banana waste and offering a gluten-free alternative in the food industry. The present study explores the feasibility of a composite mixture of GBF and JSF as a nutritious option for preparing breakfast flakes, evaluating their proximate composition and functional properties. The flakes were formulated with 15% JSF and varying ratios of GBF and Corn flour (w/w); T1 (50:35) T2 (60:25), T3 (70:15), T4 (80:5), and a control sample (C) with 100% corn flour. Physico-functional properties of GBF and JSF, proximate composition and antioxidant activity of the breakfast flakes were determined using standard protocols. Sensory evaluation was conducted by a semi-trained panel using a 5-point hedonic scale. The physico-functional data suggested JSF had higher water absorption (24.66±0.58%) and oil absorption (52.33±2.52%) than GBF (5.08±0.38%, 6.14±0.38%). In the sensory data, the T3 sample exhibited the highest scores for taste (4.03±0.91), texture (3.73±0.87), crunchiness (4.03±0.13), and overall acceptability (3.90±0.95). Incorporating GBF and JSF positively impacted nutritional content, with T4 showing the highest crude fibre levels (5.86±0.11%) and total ash content (2.84±0.05%). Additionally, antioxidant activity was significantly elevated (P&lt;0.05) in flakes with GBF and JSF, with the highest observed in T4 (19.70±0.14%) followed by T3 (18.66±0.13). Storage study data confirmed a shelf-life of 4 weeks for the developed flake samples without added preservatives at room temperature. Therefore, the present study demonstrates the potential of GBF and JSF as a healthy alternative in the bakery industry, offering both nutritional benefits and favourable sensory properties.

The addition of ascorbic acid improves the microbiological quality and shelf life of Atlantic mackerel (&lt;i&gt;Scomber scombrus&lt;/i&gt;) fillets stored in ice

Ascorbic acid is one of the most promising additives for improving fish shelf life, but studies on its antimicrobial activity during product storage are limited. In this experiment, we assessed the effect of ascorbic acid on the preservability of Atlantic mackerel (Scomber scombrus) fillets during storage in ice. Fillets were treated to reach an ascorbic acid concentration of 0.30 mg/g (T1) and 0.15 mg/g (T2). Samples were stored in ice at 0°C and analyzed at 0, 2, 7, 9, and 14 days for total viable count, specific spoilage organisms (SSOs), Pseudomonas spp., potentially histamine-producing bacteria (HPB), and Enterobacteriaceae. Growth curves were constructed using the DMfit tool of Combase, and the shelf life was estimated in relation to microbiological limits reported in the literature. Sensory evaluation was performed using a quality index method scheme. At 7 and 9 days of storage, treated samples exhibited bacterial counts from 0.5 to 1.7 Log CFU/g lower than controls, with a logarithmic reduction proportional to the additive concentration. The antimicrobial action appeared to diminish after 14 days. A consistent effect was observed for potentially HPB, with counts of 1.7 Log CFU/g lower in T1 samples compared to controls at 9 days. The moderate effect on SSOs limited the estimated shelf life, as the critical limit was reached after 7 days and 8.4 days (T1 and T2). Ascorbic acid positively impacted the microbiological characteristics of mackerel fillets. Further investigation into the decay dynamics of the additive in fish products during storage is advisable.

Effects of sub-solvus ageing on the tensile and creep properties of a new cast nickel-based superalloy

For nickel-based superalloys with medium volume-fraction γʹ phase (20 %–40 %), dual or multi-stage aging treatments are usually conducted to generate a microstructure containing the multimodal distribution of γʹ for a balance of strength and plasticity. In the present study, the microstructure and high-temperature properties of a novel cast nickel-based superalloy K4800 were investigated after being subjected to three heat treatments (HT) procedures, namely HT1: 1180 °C/4 h + 1090 °C/2 h + 800 °C/16 h, HT2: 1180 °C/4 h + 1060 °C/2 h + 800 °C/16 h and HT3: 1180 °C/4 h + 800 °C/16 h. It was found that the sub-solvus aging treatments at 1090 and 1060 °C precipitated sub-micron-sized (∼300 nm) primary γʹ phase which enhanced the ductility during 800 °C tensile (the total elongation of T1, T2, and T3 samples were 6.75 %, 7.3 %, and 3.25 %, respectively) without evidently impairing the strength. After careful microstructure observation and deformation mechanism analysis, the enhancement of elongation was rationalized that the precipitation of the sub-micron-sized primary γʹ phase decreased the volume-fraction and size of the nanometer-sized γʹ phase which was precipitated at 800 °C, and simultaneously, promoted the dislocation movement by suppressing the non-planar slip. However, an excessive amount of the sub-micron-sized primary γʹ phase led to a faster ripening process of the nanometer-sized γʹ during creep, which decreased the creep life at 800 °C/430 MPa (T1: 125 h, T2: 199 h, and T3: 198 h). Based on this, we monitored the number density of nanometer-sized γʹ phase coexisting with different amounts of large γʹ during creep. An area fraction less than 7 % of the sub-micron-sized γʹ phase was considered to have little detrimental effect on the creep life of K4800 alloy, which corresponded to a sub-solvus temperature range about 1080–1090 °C.

The immunomodulatory impacts of endocrine therapy on host immunity in early-stage hormone receptor positive, HER2 negative breast cancer.

572 Background: Endocrine therapy (ET) reduces mortality in hormonally driven breast cancer (BC). The impacts of ET on the estrogen receptor are well known; however, there is limited data on the impact of ET on the host's immune system. This project aims to elucidate the potential immunomodulatory effects of ET-induced estrogen deprivation in HR+/HER2 negative early-stage breast cancer patients (pts) as well as baseline differences between BC pts and healthy controls (HC). Methods: Pts with Stage I-III HR+/HER2- BC initiating ET after completion of local regional therapy and/or chemotherapy were enrolled (NCT03719495). Whole blood, E1 (estrone) and E2 (estradiol) levels were collected at baseline (T1), 4 weeks (T2), 12 weeks (T3) and 24 weeks (T4) after ET initiation. 5 cohorts were enrolled: premenopausal and postmenopausal HCs, premenopausal pts on tamoxifen (TAM) or aromatase inhibitors (AIs) plus ovarian suppression, postmenopausal pts on AIs. Bulk RNA sequencing of whole blood was performed using NuGEN mRNA-seq on the Illumina Nova-seq platform. Differential expression analysis was performed using DESeq2 between HC and ET groups at T1, and between T1 and T4 within BC pts. The Benjamini-Hochberg method was used to adjust the P values for multiple comparisons. A curated set of 192 immune related gene signatures was used in the gene signature analysis. For each signature/module, a score was computed by taking the median of the expression values from all genes in each gene signature. CIBERSORT analysis was performed using the LM22 signature gene file and 100 permutations. Paired t-test was performed to compare T1 and T4. Results: Between 2019 and 2023, 64 pts were enrolled. Pts with missing samples or prior chemotherapy were excluded in this analysis. T1 samples of 25 pts with BC versus 14 HCs revealed marked changes in circulating immune cell (IC) composition with 537 DEGs (p value &lt; 0.05). Circulating ICs in BC pts had marked upregulation of genes associated with oxidative phosphorylation, adipogenesis, interferon alpha, reactive oxygen species compared to HCs. Immune cell composition was inferred by CIBERSORT analysis which suggests that BC pts had fewer circulating naïve B cells (p=0.048) whereas regulatory T cells (p=0.025) and activated NK cells (p=0.0039) were higher in BC pts than HCs at T1. BC pts with reductions in E1/E2 levels 6 months after ET had 77 DEGs in PBMCs between T1 and T4. Gene signature analysis shows that B cells and NK cells (p=0.0077) signatures are elevated post ET whereas a reduction of neutrophil signature (p=0.0034) was noted. Conclusions: There are significant differences in the baseline IC repertoire/activity in BC pts compared to HCs. The reduction in B cell activity/number and an increase in regulatory T cell number suggests a possible compromised immune system in BC pts. After surgery and 6 months of ET, these suppressed immune phenotypes reversed. Clinical trial information: NCT03719495 .

Thermoluminescence characterization of flooring tiles from Malaysia for potential use in retrospective dosimetry

In the event of natural disasters or technical malfunctions at nuclear sites, the surrounding media, including individuals, may be subject to unforeseen radiation exposure. Thus, it is not possible to determine the precise level of radiation exposure that each person has received in this scenario. To handle such a problem, the use of retrospective dosimetry can offer an accurate evaluation of the radiation dose acquired as a result of such incidents. This information enables the development of suitable remedial actions. Normally, in the region that has been impacted, several artifacts can serve as natural dosimeters. The study involved the utilization of locally available seven different brands of flooring tiles in Malaysia for the possibility of being used in retrospective dosimetry through exposures to 60Co gamma-rays within a range of 10–200 Gy. To understand the luminescence features of this material, a convenient thermoluminescence (TL) technique has been employed. The dosimetric features of the samples were investigated, including their effective atomic numbers (Zeff), reproducibility, glow curve, dose response, sensitivity, linearity index, energy dependency and fading, which is representative of the usual TL method. Additionally, by analyzing the glow curves of the irradiated samples together with the peak shape method and the initial rise methods, several kinetic parameters, including the kinetic orders (b), activation energy (E) or trap depth, frequency factor (s) or escape probability, and trap lifetime (τ) were calculated. Different kinds of tiles exhibit distinct scenarios in terms of kinetic characteristics. T1 sample exhibits the highest degree of sensitivity and a highly linear dose response among the tile samples that were evaluated. Also, T1, T3, and T5 samples show minimal loss of TL intensity (∼38–39%) one year after irradiation, making them ideal for use as a retrospective dosimeter.

Cancer associated macrophages in blood correlates with poor outcomes in patients with clinically resectable disease in gastrointestinal cancers.

e15052 Background: Gastrointestinal (GI) cancers, i.e., pancreatic (PC), colorectal (CC), and esophageal (EC), are known to be both highly aggressive and clinically underdiagnosed, resulting in high rates of relapse after standard neoadjuvant therapy with curative intent resection. While cancer biomarkers (i.e. CEA, CA19-9) can aid in identifying early relapse post-resection, they lack accuracy and do not aid in identifying underdiagnosed patients. Recently, a circulating phagocytic macrophage engorged to ≥50 µm, i.e. cancer-associated macrophage-like cells (CAMLs), were identified in GI cancers and associated with highly aggressive cancer subtypes. However, monitoring CAMLs pre- and post-surgery and their clinical value in minimal residual disease remains unexplored. In this pilot study, we prospectively evaluated the blood of n = 133 patients with GI cancer pre- and post-surgery to assess CAMLs in resectable disease, and their association with progression free survival (PFS), relapse free survival (RFS), and overall survival (OS). Methods: In a single-blind multi-institutional prospective pilot study, we recruited n = 133 GI patients with newly diagnosed clinically resectable disease (n = 118), or with metastatic disease scheduled for curative resection post systemic therapy (n = 15). Whole peripheral blood (7.5mL) was drawn prior to neoadjuvant chemotherapy and/or radiation therapy at T1, and a post-surgery sample (T2) was obtained. CAMLs were filtered, stained with cytokeratin &amp; CD45, and enumerated for CAML engorgement of ≥50 µm, as previously described. Cox proportional regression hazard ratios for PFS, RFS &amp; OS using univariate and multivariate analyses were run over 96 months. Results: Of the patients with clinically resectable EC (n = 36/133), PC (n = 75/133) or CC (n = 22/133), 32% of patients (n = 43/133) did not undergo curative resection. T1 samples drawn pre-surgery were available from n = 105 patients and T2 samples drawn post-surgery were available from = 52 patients. Engorged ≥50 µm CAMLs were found in 51% (n = 53/105) of T1 patients and were significantly associated with worse PFS (HR = 3.4 CI95% 2.0-5.8, p &lt; 0.001) and worse OS (HR = 63.5 CI95% 2.0-6.1, p &lt; 0.001), compared to patients with &lt; 50 µm CAMLs. Additionally, engorged ≥50 µm CAMLs were found in 60% (n = 31/52) of T2 patient samples and were associated with significantly worse RFS (HR = 5.7, CI95% 2.4-13.2, p &lt; 0.001) and OS (HR = 5.3, CI95% 2.2-13.3, p &lt; 0.001) compared to patients with &lt; 50 µm CAMLs. Conclusions: These data suggest that GI cancer patients with CAMLs ≥50µm prior to curative intent resection have high rates of progression, and their presence post surgery may identify patients with residual disease that are likely to relapse. Given these results, more refined studies should evaluate CAMLs ability to identify such at risk patients who may benefit from alternative therapeutic approaches.

Influence of thermocycling on the functional properties of the NiTi alloy produced by wire arc additive manufacturing

The influence of the thermocycling under a stress of 200 and 300 MPa on the martensitic transformation temperatures, recoverable and irreversible strain was study in the NiTi samples produced by wire arc additive manufacturing (WAAM). Two types of samples were used: the T1 sample including the Ti-rich NiTi and Ni-rich NiTi layers in the working length and the T2 sample including only the Ni-rich NiTi layers. It was found that the variation in the martensitic transformation temperatures on thermal cycling depended on the chemical composition of the layer. The transformation temperatures decreased on thermal cycling in the Ti-rich NiTi layer (in T1 sample), whereas they were constant in the Ni-rich NiTi layers (in T1 and T2 samples). The recoverable strain in both samples did not change during thermal cycling. The irreversible strain was found in the T1 sample regardless of stress acting on thermal cycling, while T2 sample showed the plastic strain only in the first cycle under 300 MPa. It was shown that the difference in a functional behavior of the T1 and T2 samples on thermocycling was due to that the T1 sample contained the Ti-rich NiTi layer, that was characterized by a low dislocation slip limit contrary to the Ni-rich layers which were hardened by the Ni4Ti3 precipitates.

Development of Nutritious Biscuits for Gluten Intolerance Patients

Background: The rising incidence of celiac disease and gluten intolerance has led to an increasing demand for gluten-free products. Tapioca and corn flour are promising alternatives to wheat flour due to their gluten-free nature and beneficial nutritional properties. Objective: This study aimed to develop and evaluate the sensory characteristics of gluten-free biscuits made from tapioca and corn flour, individually and in combination, compared to a control sample made from wheat flour. Methods: This study was conducted at the National Institute of Food Science and Technology, University of Agriculture Faisalabad. Ingredients such as wheat flour, corn flour, tapioca flour, sugar, butter, eggs, vanilla extract, baking powder, and salt were procured from the local market. Four samples were prepared: a control sample (T1) with wheat flour; T2 with corn flour; T3 with tapioca flour; and T4 with a blend of corn and tapioca flours. The specific quantities of ingredients were as follows: T1 (250g wheat flour), T2 (250g corn flour), T3 (250g tapioca flour), and T4 (125g corn flour + 125g tapioca flour). Biscuits were prepared by mixing dry ingredients, followed by liquid ingredients, to form a smooth dough, which was then shaped and baked at 180°C for 20-25 minutes. Sensory evaluation involved consumer effective testing with 50 untrained panelists using a 9-point hedonic scale, and quantitative descriptive analysis (QDA) with 10 trained panelists using a 15-point scale. Data were analyzed using SPSS version 25 and Microsoft Excel for graphical representation. Ethical approval was obtained in accordance with the Helsinki Declaration, and informed consent was obtained from all participants. Results: In the consumer effective test, the control sample (T1) received ratings of 7.89 for color, 7.10 for flavor, 7.85 for texture, 7.9 for aroma, and 7.68 for overall acceptance. The T2 sample showed ratings of 7.78 for overall acceptance, 7.55 for color, 7.95 for flavor, 7.65 for texture, and 7.98 for aroma. The T3 sample scored 7.62 for overall acceptance, 7.55 for flavor, 7.8 for texture, and 8.0 for aroma. The T4 sample, with the blend of corn and tapioca flour, displayed the highest scores: 8.5 for color, 8.15 for flavor, 8.1 for texture, 7.95 for aroma, and 8.17 for overall acceptance. In the QDA, the control sample (T1) received scores of 9.81 for overall acceptability, 8.55 for flavor, 9.5 for texture, 10.5 for aroma, and 11 for taste. The T2 sample scored 10.08 for overall acceptability, 9.5 for color, 9.7 for flavor, 9.1 for texture, 11 for aroma, and 11.1 for taste. The T3 sample had scores of 10.32 for overall acceptability, 9.0 for color, 11.1 for flavor, 9.6 for texture, 11 for aroma, and 10.9 for taste. The T4 sample achieved the highest scores in QDA: 11.18 for overall acceptability, 10 for color, 13 for flavor, 10 for texture, 11.9 for aroma, and 11 for taste. Conclusion: The study demonstrated that gluten-free biscuits made from a blend of tapioca and corn flours (T4) had superior sensory attributes compared to those made from either flour alone and the control sample made from wheat flour. This suggests that tapioca and corn flour blends could be promising alternatives in the development of gluten-free products. Further research is recommended to explore the application of these flours in other gluten-free food products.

#2402 Reproducibility of memory B-cell and HLA antibody detection in patients on kidney transplant waiting list

Abstract Background and Aims Detection of HLA antibodies (HLA-Abs) impacts on the access and outcomes of kidney transplantation (KT). Currently, evaluation of HLA-Abs in serum is performed using single antigen bead assays (SAB). The analysis of circulating HLA-specific memory B cells (mBc), capable of differentiating to HLA-Abs secreting cells, was conceived with the aim of complementing the study of the HLA-Abs found in serum, but there is debate regarding the representativeness and reproducibility of the test. Thus, we studied HLA-Abs generated by mBc from patients on the waiting list (WL) at two different time points without sensitizing events in between and compared them with HLA-Abs detected in serum. Method We selected 7 patients on the WL for a first KT with a cPRA of 3%-100%: 4 had pregnancies and 2 had blood transfusions as known sensitizing events. We collected blood and serum samples at 2 time points separated by an average of 3.5 months (T1, T2). We cultured polyclonally activated PBMCs and isolated and concentrated IgG from the supernatant (SN) to study HLA-Abs. We tested SN and sera with SAB assays. Beads with MFI &amp;gt;750 were considered positive. Results SAB serum study (Fig. 1a). Eighty-two percent of the specificities found at T1 were also detected at T2, and 97% of those identified at T2 were also observed at T1. Thus, there was good concordance between T1 and T2 samples (κ = 0.869, Cohen) and the MFI the specificities correlated (p &amp;lt; 0.0001, Spearman). SAB SN study (Fig. 1b). Forty-one percent of the positive specificities detected at T1 were also identified at T2, and 59% of the positive specificities found at T2 had been seen at T1. Thus, there was a moderate concordance between T1 and T2 samples (κ = 0.454, Cohen) and the MFI of the specificities still correlated (p &amp;lt; 0.0001, Spearman). Only 42% and 35% of the positivities found in serum were also detected in the SN at T1 and T2, respectively. However, we found 9 positivities unique to the SN T1 (100% HLA-ABC) in 2 patients with cPRA&amp;gt;80% and we also identified a total of 20 positivities unique to the SN at T2 (45% HLA-ABC, 55% HLA-DPDQDR) in 3 patients with cPRA&amp;gt;80%. Conclusion The study of the degree of sensitization with SAB serum analysis remains stable between samples along time, while the mBc analysis shows less reproducibility. We identified a higher number of HLA-Ab in the study of serum compared with the study of the SN of mBc; however, we found some HLA-Abs unique to the SN. We suggest that, probably due to the small number of circulating mBC, increasing the number of samples tested for mBc improves the representativeness of the test.

Effect of Heat Treatment on the Material Property and Cell Viability of Wire Arc Additively Manufactured Ti6Al4 V.

Wire arc additive manufacturing (WAAM) holds promise for producing medium to large industrial components. Application of WAAM in the manufacturing of biomedical materials has not yet been evaluated. The current study addresses two key research questions: first, the suitability of the WAAMed Ti6Al4V alloy for biomedical applications, and second, the effect of Ti6Al4V's constituents (α and β phases) on the cell viability. The WAAMed Ti6Al4V alloy was fabricated (as-deposited: AD) using a metal inert gas (MIG)-based wire arc system using an in-house designed shielding chamber filled with argon. Subsequently, samples were subjected to solution treatment (950 °C for 1 h), followed by aging at 480 °C (T1), 530 °C (T2), and 580 °C (T3) for 8 h and subsequent normalization to ambient conditions. Microstructural analysis revealed ∼45.45% of α'-Ti colonies in the as-deposited samples, reducing to 23.26% postaging at 580 °C (T3). The α-lath thickness and interstitial oxygen content in the sample were observed to be proportional to the aging temperature, peaking at 580 °C (T3). Remarkably, during tribocorrosion analysis in simulated body fluid, the 580 °C-aged T3 sample displayed the lowest corrosion rate (7.9 μm/year) and the highest coefficient of friction (CoF) at 0.58, showing the effect of increasing oxygen content in the alloy matrix. Cell studies showed significant growth at 530 and 580 °C by day 7, correlated with higher oxygen content, while other samples had declining cell density. Additionally, optimal metallurgical property ranges were identified to enhance the Ti6Al4V alloy's biocompatibility, providing crucial insights for biomedical implant development.

Abstract 3690: Monitoring prostate specific membrane antigen or androgen receptor expression on circulating stromal cells in advanced prostate cancer patients and their correlation with patient response

Abstract Background: Metastatic Prostate cancer (mPCa) mortality rates are high despite numerous treatment options against various mechanisms of action, including targeted therapies, i.e. prostate specific membrane antigen (PSMA)-targeted therapy (Pluvicto) or anti-androgen receptor (AR) (Enzulamide, Biculamide). Further, tumor biomarkers (i.e. PSMA and AR) can change over time and with resistance mechanisms that develop after treatments. This suggests that primary biopsies may not represent later PCa tumors necessitating a way to identify patients that may respond to later line therapies. Blood based biopsies can be used to monitor PCa treatment, which can include circulating tumor cells (CTCs) and the newly discovered cancer associated macrophage like cells (CAMLs), a cancer specific circulating phagocytic stromal cells. Interestingly, while PSMA &amp; AR have been identified in CTCs, they have not been evaluated in CAMLs, nor have PSMA &amp; AR expressions in CTCs &amp; CAMLs been evaluated in anti-AR therapy settings. We measured CAMLs &amp; CTCs in mPCa to evaluate their PSMA &amp; AR expression, including in patients treated with anti-AR therapy. Methods: We evaluated CAMLs &amp; CTCs in a multi-institutional prospective pilot study using n=30 mPCa patients with progressive disease, prior to starting a new line of therapy (T0). Whole peripheral blood (7.5mL) was filtered for CAMLs &amp; CTCs and stained for PSMA (n=15) &amp; AR (n=15). In addition, 15 patients were treated with anti-AR therapy as standard of care and responses by PET/CT were compared against AR expression. When possible, follow up samples (T1) were also procured. Results: CTCs were identified in 16% (n=5/30) of patients and CAMLs were identified in 96% (n=29/30), with 53% of patients having hyperengorged CAMLs ≥50 μm (n=16/30). CTCs in patients were not statistically significant for worse outcomes PFS HR= 0.1748, p=0.546 nor OS (HR= 1.303, p=0.865), nor were CAMLs prognostic for PFS or OS. However, patients with ≥50 μm CAMLs did have significantly worse PFS (HR= 5.9, CI95% 2.2-16.3, p=0.001) and worse OS (HR= 3.1, CI95% 1.3-7.1, p=0.016). Further, PSMA CAML/CTC expression was high in 60% (n=9/15) of patients and AR CAML/CTC expression was high in 53% (n=8/15). Of the n=15 patients who received anti-AR therapy after T0, high AR expressing CAMLs/CTCs (n=8/15) had a ~250% better mPFS of 10.3 months (mons) (progressing at 2.2-32.6 mons) vs low AR CAMLs/CTCs mPFS of 4 mons (progressing at 0.2-7.2 mons). Additional mOS analysis from T1 samples and 2-year survival is ongoing. Conclusion: We utilized liquid biopsies to monitor the expression of two common cell surface receptors (PSMA &amp; AR) in CAMLs and CTCs in mPCa. Further, in a small subset of patients treated with AR therapy, high AR CTC/CAML expression appeared to correlate with better response rates, though larger prospective studies are needed. Citation Format: Dimpal M. Kasabwala, Raymond C. Bergan, R. Katherine Alpaugh, Daniel C. Danila, Tuan L. Chuang, Brenda Y. Hurtado, Daniel L. Adams. Monitoring prostate specific membrane antigen or androgen receptor expression on circulating stromal cells in advanced prostate cancer patients and their correlation with patient response [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 3690.

Abstract 7531: Mechanisms of response to anti-PD1 (PD1) combined with Ipilimumab (IPI) in patients (pts) with PD1-resistant metastatic melanoma (MM)

Abstract Background: Approximately 1 in 3 pts with PD1-resistant MM will benefit from IPI+PD1, but the role of adding IPI in this context is unclear. We sought to (a) identify biomarkers and (b) determine mechanisms of response to IPI+PD1 in PD1 resistant pts. Methods: Pts with PD1-resistant MM subsequently treated with IPI+PD1 were included. We performed multiplex immunofluorescence (mIF), imaging mass cytometry (IMC) and RNA sequencing (RNAseq) on melanoma samples, and cytometry by time of flight (CYTOF) on peripheral blood mononuclear cells (PBMCs), at 3 timepoints (before PD1 [T1], before IPI+PD1 [T2], and after IPI+PD1 [T3]). Results: 34 pts with PD1-resistant MM (82% innate resistance) who received IPI+PD1 were included: 16 (47%) responders and 18 (53%) non-responders to IPI+PD1. Median age was 66yo (37-86) and 19 (56%) were male. Six pts (18%) were ECOG PS ≥1, 18 (53%) were stage M1C/D and 11 (32%) had elevated LDH at the start of IPI+PD1.Gene expression analysis (RNAseq) on T1 and T2 tumor samples revealed higher expression of cancer testis antigens (CTA; MAGE-A3 and MAGE-C1; p&amp;lt;0.05) and immune regulatory genes (ILR2; p&amp;lt;0.2), but lower expression of fibrosis regulation genes (FABP3 and MMP7; p &amp;lt; 0.2) in responders vs non-responders. We then examined whether the addition of IPI led to expansion of CTA-specific T cells, and CyTOF analysis of MAGE-A3 tetramers on PBMCs showed a decrease in peripheral MAGE-A3-specific CD8+ T cells with PD1 (T1 to T2; p=0.03); and expansion of this T-cell subset with subsequent IPI+PD1 (T2 to T3) was seen in 7/12 HLA-A2+ pts. Further analysis on PBMC longitudinal samples showed an increase in regulatory T cells (Tregs) in responders (p=0.04), between T1 and T3, and a decrease in % of TCF7+ CD8+ &amp; CD4+ T cells in non-responders (p&amp;lt;0.05) with the addition of IPI (T2 to T3). On baseline (T1) tumor samples (mIF), IPI+PD1 responders had a higher % of stem cell-like TCF7+ CD8+ T cells (p=0.0004), lower % of well-differentiated Tbet+ CD8+ T cells (p=0.07), and higher % of CD4+ T cells (p=0.006), compared with non-responders; but no difference in Tregs. On longitudinal tumor samples, there was a decrease in the % of stem cell-like TCF7+ CD8+ T cells but an increase in the % of tumor-reactive CD39+ CD103+ CD8+ T cells with PD1 (T1 to T2; p&amp;lt;0.05), mainly seen in responders (p=0.06). There were only 3 T2 to T3 IHC paired samples, precluding further analysis. IMC analysis on longitudinal samples (T1, T2 and T3) is underway to characterize the interaction between Tregs and T effector cells. Conclusions: We identified potential mechanisms of response to IPI+PD1 in PD1-resistant MM pts. Our data suggests that response to IPI+PD1 requires baseline stem cell-like TCF7+ T cells with the potential to differentiate to an effector phase (CD39+ CD103+ CD8+ T cells), and that IPI facilitated expansion of antigen-specific T cells in a CTA-expression context, that PD1 alone is unable to do. Citation Format: Ines Pires da Silva, Jordan Conway, Jarem Edwards, Felix Marsh-Wakefield, Camelia Quek, Angela Ferguson, Peter Johnansson, Matteo Carlino, Alexander Menzies, James Wilmott, Umaimainthan Palendira, Richard Scolyer, Georgina Long. Mechanisms of response to anti-PD1 (PD1) combined with Ipilimumab (IPI) in patients (pts) with PD1-resistant metastatic melanoma (MM) [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 7531.

Effect of gutting on microbiological, chemical and sensory properties of Urechis unicinctus stored in ice and at chill temperature

This study investigates the effect of gutting on quality characteristics of Urechis unicinctus, stored in ice and at chill temperature (4℃), in terms of sensory assessment, total volatile basic nitrogen, biogenic amines and microbial changes. The gutted and ungutted groups were kept in ice (T1-gutted, T2-ungutted) and at chill (4℃) temperature (T3–gutted, T4-ungutted). The study revealed that gutting worsened the sensory quality (as indicated by hardness, cohesiveness, springiness and chewiness) of all samples and that samples kept at 4°C scored lower than those kept in ice. Total Volatile Basic Nitrogen registered the highest values in gutted samples, exceeding the acceptable limit after 1 day for the T3, and 12 days for T1 samples, respectively. For ungutted samples the threshold value of 30 mg/L has been exceeded after 4 days for T4, and 14 days for T2 samples. In the present study, eight biogenic amines were analyzed; tryptamine and histamine were not detected all the time. The total concentration of biogenic amines in gutted U. unicinctus was significantly higher in comparison with ungutted groups starting from the 8th day, for samples stored in ice, and from the 3rd day for samples kept at 4℃ (p &lt; 0.05). However, the concentration of biogenic amines did not reach the allowable limit, even the samples were unacceptable due to high values of total viable count. Therefore, limits of biogenic amines available for fish are not suitable for U. unicinctus. Sensory scores were significantly correlated with Total Volatile Basic Nitrogen and Total Viable Count in all four groups (p &lt; 0.05). Combined results of this study indicated that shelf life of T1, T2, T3 and T4 were 6, 7, 1 and 4 days, respectively. Gutting had a little effect on the storage of U. unicinctus stored in ice, but obviously shortened the shelf life of samples when stored at 4℃.