Single Lot Research Articles

Post-irradiation darkening model for EBT-3 films characterized using a single lot calibration approach

Objective.In this study, we present a model to correct the progressive post-irradiation darkening of EBT3 films. The model allows for a clinical use of EBT3 using application and calibration films scanned with different post-irradiation times.Approach.The model is a post-irradiation time- and dose-dependent power-law function, projecting the scanned transmittance of application films to the transmittance matching the same post-irradiation time of calibration films. The model was characterized for two EBT3 production lots within the dose range 0.1-12.8 Gy. A first characterization was performed utilizing calibration films scanned repeatedly for 54 d post-irradiation (lot 1), while a fast re-characterization of a second lot used three post-irradiation times (lot 2). For a long-term validation of the model, 16 film strips were irradiated at 2 Gy on different time points starting from the day of film calibration up to 43 d afterwards (lot 1). For the multiple-dose validation of the model, 8 strips were irradiated with dose levels ranging 0-12 Gy deposited 25 d after the calibration (lot 2). As a proof of principle, the model was applied to four clinical patient-specific quality assurance film measurements with prescribed dose/fraction ranging 2.66 Gy-8 Gy.Main results. The post-irradiation transmittance decreased for higher doses up to -2.5% at 12.8 Gy, and 54 d post-irradiation. With a lot-specific model correction, the mean dose accuracy of validation strips that ranged from initial -3.4% (triple-channel) and -9.90% (blue-channel) reduced to within 3.0% (all colour channels) for doses above 1 Gy. The median dose difference with the planned dose improved from -3.5% to -1.1%, and the 3%/2 mm local gamma ranged from (48.5-92.5)% to (81.2-99.2)%.Significance.A film darkening model corrects the transmittance of EBT3 films and increases the flexibility of existing dosimetry protocols. The correction ensures dose accuracies within 3%.

B-143 Using Big Data Analytics to Measure the Multi-year Consistency of Sigma Metrics for Vitros Oncology Assays

Abstract Background Sigma metrics assess lab quality by incorporating precision, accuracy and total error allowed (TEa). Sigma metrics report quality on a 0-6 scale, with 6 representing World-Class quality. Due to logistical constraints, Sigma metrics are often calculated with limitations such as a small number of analyzers, single reagent lot or short duration. QuidelOrtho developed a novel approach using big data analytics for the objective, automated calculation of Sigma metrics across thousands of Vitros analyzers running in real lab conditions. This approach was previously used to report the Sigma metric performance for 115 assays using data from 2022. Of particular interest is the consistency of quality levels for Vitros oncology assays assessed in the study. The aim of this analysis was to examine Sigma metrics for Vitros oncology assays and compare Sigma metrics to data from 2022. Methods The methods follow the previous approach using the same Python algorithm to extract worldwide QC data from Jan. to Nov. 2023 (inclusive). A QC fluid lot from the study period was selected for each analyte to align with the fluid level reported in the 2022 study, which was based on medical relevance. Metrics were calculated for each set of QC data that originated from a single analyzer, reagent lot and calibration curve; the median Sigma metric is reported in the Results. Proficiency testing limits were utilized as TEa limits where available, with preference to CLIA 2024 limits. Results The following assays reported Sigma metrics as >6 using both 2022 and 2023 data: alpha-fetoprotein, cancer antigen 125, carcinoembryonic antigen, prostate-specific antigen (PSA), free PSA, IgA, and IgG. Assays with higher metrics using 2023 data: cancer antigen 19-9 (5.0 vs >6) and IgM (5.3 vs >6). Assays with lower metrics using 2023 data: cancer antigen 15-3 (4.8 vs 4.4). Conclusions Vitros oncology assays report similar performance for 2022 and 2023. All Vitros oncology assays assessed report Sigma metrics of 4 or greater, indicating they achieve results within the corresponding allowable error.

B-142 Using Big Data Analytics to Measure the Multi-year Consistency of Sigma metrics for Vitros Cardiac Assays

Abstract Background Sigma metrics assess lab quality by incorporating precision, accuracy and total error allowed (TEa). Sigma metrics report quality on a 0-6 scale, with 6 representing World-Class quality. Due to logistical constraints, Sigma metrics are often calculated with limitations such as a small number of analyzers, single reagent lot or short duration. QuidelOrtho developed a novel approach using big data analytics for the objective, automated calculation of Sigma metrics across thousands of Vitros(R) analyzers running in real lab conditions. This approach was previously used to report the Sigma metric performance for 115 assays using data from 2022. Of particular interest is the consistency of quality levels for Vitros cardiac assays assessed in the study. Methods The methods follow the previous approach using the same Python algorithm to extract QC data from Jan. to Nov. 2023 (inclusive). A QC fluid lot from the study period was selected for each analyte to align with the fluid level reported in the 2022 study, which was based on medical relevance. Metrics were calculated for each set of QC data that originated from a single analyzer, reagent lot and calibration curve; the median Sigma metric is reported in the Results. Proficiency testing limits were utilized as TEa limits where available, with preference to CLIA 2024 limits. Results The following assays reported Sigma metrics as >6 using both 2022 and 2023 data: apolipoprotein A1, apolipoprotein B, CK-MB, dLDL, LDH, myoglobin, NT-proBNP, and triglycerides. Assays with higher metrics using 2023 data: CK (5.2 vs 5.5), dHDL (5.8 vs >6), hsTnI (4.1 vs 4.3), and NT-proBNP2 (4.2 vs >6). Assays with lower metrics using 2023 data: cholesterol (5.8 vs. 5.3) and homocysteine (>6 vs 5.9). CKMB2 (>6) is a newer assay that did not have metrics reported in 2022. Vitros cardiac assays report metrics within 0.5 Sigma of the 2022 metrics. An exception is NT-proBNP2, which changed from 4.2 to >6. However, the 2023 analysis has almost double the number of Vitros analyzers (51 vs 90), representing new use of NT-proBNP2. Conclusions Vitros cardiac assays report consistent metrics when using 2023 data compared to 2022. All Vitros cardiac assays assessed report Sigma metrics of 4 or greater, indicating they achieve results within the corresponding allowable error.

A-110 Analytical Performance Evaluation of Vitros® Immunodiagnostic Products Intact PTH II Assay

Abstract Background Intact parathyroid hormone (iPTH) is indicated to aid in the diagnosis of hyperparathyroidism, hypoparathyroidism, differential diagnosis of hypocalcemia or hypercalcemia and for intraoperative measurement of iPTH levels. Studies were conducted to evaluate the analytical performance of the Vitros Immunodiagnostic Products Intact PTH II (Vitros iPTH II) assay by determining limit of detection (LoD), limit of quantitation (LoQ), and precision. A method comparison of the Vitros iPTH II assay against the Roche Elecsys PTH assay was also conducted. Methods The LoD and LoQ were determined consistent with CLSI EP17. Precision was evaluated consistent with CLSI EP05 using six patient pools for 20 days. Repeatability was determined using two replicates per run. Within-lab precision was determined using a single reagent lot and a single calibration. A quantitative comparison between the Vitros iPTH II assay on a Vitros 3600 system and the Roche Elecsys PTH assay on a Roche Cobas 8000 system was conducted in accordance with CLSI guideline EP09 using 206 endogenous patient samples. Samples were selected to cover the full measuring interval and were tested over a total of six non-consecutive days. Results The LoD for the Vitros iPTH II assay was determined to be 1.7 pg/mL. The claimed LoQ was determined using a precision profile approach with a %CV limit of 20% and set at 1.7 pg/mL. For mean parathyroid hormone (PTH) concentrations ranging from 15.9 to 4656 pg/mL, repeatability (within-run) percent coefficient of variation (%CV) ranged from 0.8% to 1.4% and within-lab %CV ranged from 1.5% to 3.0%. The method comparison of the Vitros iPTH II assay versus the Roche Elecsys PTH assay yielded a correlation coefficient of 0.991 and a Passing-Bablok regression slope of 1.01 and intercept of 0.3. Conclusions The Vitros iPTH II assay demonstrated acceptable analytical performance in these studies, with high correlation and close agreement to the Roche Elecsys PTH assay.

B-144 Sigma Metrics for Vitros Reproductive Endocrinology Assays Across Multiple Fluid Levels

Abstract Background Sigma metrics assess laboratory quality by incorporating precision, accuracy and total error allowed (TEa). Sigma metrics report quality on a scale of 0 to 6, with 6 representing the World-Class quality. QuidelOrtho developed a novel approach using big data analytics for the objective and automated calculation of Sigma metrics across thousands of Vitros® Analyzers running in real laboratory conditions. This approach was previously used to report the Sigma metric performance for 115 assays using data from 2022. A limitation of that study is the metrics were reported for a single fluid that was selected for medical relevance. However, reproductive endocrinology assays have complex medical utilization and there is a benefit to understanding Sigma metrics at multiple levels. The aim of this poster is to provide additional data on the Sigma metrics for reproductive endocrinology assays. Methods The methods follow the previous approach using the same Python algorithm to extract QC data from Jan. to Aug. 2022 (inclusive). Up to seven QC fluid lots were selected for each analyte. Metrics were calculated for each set of QC data that originated from a single analyzer, reagent lot and calibration curve; the median Sigma metric is reported in the Results. Proficiency testing limits were utilized as TEa limits where available, with preference to CLIA 2024 limits. Results Sigma metrics are reported for multiple fluids for the following assays: estradiol, follicular stimulating hormone, luteinizing hormone, progesterone, prolactin, testosterone, human chorionic gonadotropin, and thyroid stimulating hormone. All fluids examined report Sigma metrics greater than 4, which is considered “Good” performance. An exception is luteinizing hormone which has Sigma metrics = 1.9 at the lowest reported concentration. Conclusions Vitros reproductive endocrinology assays report strong Sigma metrics across multiple concentrations, indicating they are highly capable of achieving results within the allowable error.

Ralstonia mannitolilytica infection in a tertiary care centre: an outbreak report

BackgroundIn this paper we describe an outbreak of Ralstonia mannitolilytica infection declared in our facility between January 2021 and January 2022. MethodsIn order to identify the source of the outbreak, we applied widespread epidemiological investigation and infection control measures, including device isolation, environmental sampling, and PFGE typing. ResultsThirty-six cases of R. mannitolilytica infection were identified, mostly adults (78%) and males (75%). Initially, neurological procedures were a common risk factor among cases, leading to sampling of related environmental settings. Cases with other medical procedures started to be reported. The PFGE results showed most R. mannitolilytica isolates were indistinguishable which expanded our investigation to all hospital areas. The infection source was discovered later as a single lot number of sodium chloride solution that had been used in practice. The contaminated solution was recalled from all hospital units and the findings were reported to the Saudi FDA to communicate with the manufacturer and other healthcare organizations involved. No new cases of R. mannitolilytica were identified thereafter. ConclusionOur findings indicate that identifying the source of an outbreak could be challenging. An extended incubation period might be considered to improve and accelerate the identification of R. mannitolilytica. In order to minimize similar incidents, it is essential to regularly monitor the compliance of manufacturers and suppliers with regulations related to the safety of solutions administered in medical practice.

The mangrove oyster Saccostrea palmula is an option for artisanal aquaculture development in the Mexican Central Pacific

The mangrove oyster Saccostrea palmula is a relatively small but emerging species in aquaculture, and it is necessary to resume and increase studies on its cultivation. Therefore, we carried out a study that includes three main components for developing S. palmula culture in the Mexican Central Pacific: spat collection, pre-growth phase (eight weeks), and grow-out phase (six months). We tested natural (shell strings) and artificial (coupelles) collectors placed in contrasting environmental conditions: marine (La Boquita) and estuarine (Cuyutlan). The spat was pre-grown in suspended Australian trays at the two collection sites and under in-land pond conditions. Then, it was pooled under an estuarine environment and in a single lot for the final culture phase in Juluapan lagoon (all sites in Colima, Mexico). Spat collection was higher in coupelles (858.3 ± 175.6 spat coupelle-1) than in shell strings. The number of spat collected (13,062 spat), final pre-growth size (26.5 ± 0.6 mm shell height), and pre-growth survival (94.7%) were higher in La Boquita (marine environment). The final culture phase in Juluapan (estuarine conditions) was successful, recording an average height of 54.2 ± 4.9 mm and average total weight of 30.8 ± 6.9 g after six months of culture, after spat collection and pre-growth periods. We found the largest oyster, S. palmula, ever recorded (shell height 83.3 mm and total weight 59.4 g), and we confirmed that S. palmula has potential for artisanal aquaculture and socioeconomic regional development in the Mexican Central Pacific.

Enhancing mango quality control: A novel approach to spongy tissue inspection through image clustering and machine learning models via X‐ray imaging

AbstractThe globally renowned Alphonso mango is esteemed for its exceptional flavor, fragrance, texture, and extended shelf life. However, it faces a substantial challenge with spongy tissue (ST) disorder, affecting up to 30% of a single lot. This issue leads to the rejection of entire batches during export due to delayed detection, causing significant produce loss. Additionally, the current destructive assessment method is inadequate, as physically inspecting only a few mangoes within a batch does not ensure overall quality, emphasizing the need for a non‐destructive approach. To address these challenges, this study proposes a non‐destructive approach utilizing X‐ray imaging to identify ST in Alphonso mangoes, employing machine learning (ML) for binary classification. We captured and augmented 216 X‐ray images, utilizing various augmentation techniques. K‐means clustering effectively distinguished healthy and ST‐affected areas in mango fruits. For automated classification, ML models were fine‐tuned through grid search hyperparameter optimization within a 5‐fold cross‐validation framework. The CNN model achieved impressive accuracies of 99.20% and 96.82% at threshold values of .5 and .95, respectively, on the test dataset, while the SVM model attained overall accuracy rates of 95.23% and 87.35% for the same thresholds. Our study demonstrates the potential of machine learning in X‐ray imaging for detecting internal fruit issues, improving inspection for disease and damage identification, and enhancing quality control across the food industry. Practical implications include the implementation of non‐destructive inspection methods to reduce produce loss and ensure consistent quality in mango lots. Future research could focus on validating the model's efficacy with external datasets, evaluating its robustness in detecting various internal irregularities, and exploring advancements in X‐ray imaging technology for broader applications in agriculture.Practical applicationsThe suggested non‐destructive approach provides a practical solution for quality control, especially in export scenarios. The versatility of the ML‐powered X‐ray imaging technique to examine internal irregularities in various fruits beyond mangoes broadens its potential application in the fruit industry for sorting and grading purposes. This innovation contributes to heightened consumer satisfaction, providing a cost‐effective and scalable solution for widespread implementation. In summary, the findings of the study show promise for revolutionizing mango quality control, particularly for the esteemed Alphonso variety, and propelling advancements in fruit inspection methodologies across the industry.

Parallel Algorithm on Multicore Processor and Graphics Processing Unit for the Optimization of Electric Vehicle Recharge Scheduling

Parallel Algorithm on Multicore Processor and Graphics Processing Unit for the Optimization of Electric Vehicle Recharge Scheduling

Economic Production Quantity Model Considering Rework of Imperfect Goods under Stock, Advertisement, Selling Price and Replacement Period-Dependent Demand

An economic production quantity (EPQ) system with an imperfect production model in which demand is affected by advertisement frequency, selling price, stock level and product replacement time is discussed in this paper. It is unavoidable to produce only perfect quality goods in a manufacturing operation due to real-life issues like labor problems, machinery faults, etc. Therefore, we consider the production system imperfect and manufacture a fraction of imperfect goods that depend on the production rate. All produced goods are continuously shifted to the screening center, where perfect goods are directly sold to the consumers. After the screening period, the imperfect goods are transferred to the reworking center, where some imperfect goods are reworked at a price. The remaining defective products are sold as a single lot. A profit function is developed for this proposed production inventory model, which is highly nonlinear. The total profit is maximized using the graphical method to get the optimal production quantity and total business time. The profit function’s concavity is shown graphically by using the Mathematica software. To demonstrate the model’s feasibility, numerical examples are provided. Sensitivity analysis is carried out to highlight the findings of the suggested production inventory system and depict the managerial insights.

Realising transitions for urban regeneration through statutory planning: the case of greyfield precinct renewal in suburban Melbourne

ABSTRACT The compact city agenda aims to deliver sustainable future urban form. However, as with all planning, there is contestation. While built form and housing have primacy, cities need space for hard and soft infrastructural systems, including natural systems that, until recently, have been taken for granted but are diminishing through poorly planned densification. Nowhere is this more evident than in the suburbs, where lot-by-lot redevelopment is significantly reducing open space and tree canopy. Infill developments are not producing the benefits expected from compact cities; they are increasing strain on the grey, blue, and green infrastructure. This paper highlights limited intent in local planning regulations to promote anything other than business as usual, which, in the Australian context, is single lots becoming 2–4 subdivisions. While planning schemes can regulate negative externalities, they do not consider this when taken as a whole, across a redeveloped neighbourhood predominantly consisting of housing, and are not using the redevelopment opportunities to build a more resilient future for local areas. This paper builds on the Australian greyfield literature by showing how statutory amendments can promote precinct scale development, working across multiple lots to deliver housing concurrently with greater levels of amenity for existing and future residents. KEY POINTS Infill housing development on single lots is reducing local amenity while not making the fullest use of the land for housing supply. Site assembly precincts are critical for higher yield, higher amenity, urban redevelopment. Strategic policy alone will not deliver the solutions; statutory mechanisms for doing so are critical. This paper presents a fully worked statutory response to the issue, highlighting the method for developing the scheme and the future work necessary for delivering site-assembly precincts at council level.

Standardization of flow cytometric detection of antigen expression.

Since response to antigen-based immunotherapy relies upon the level of tumor antigen expression we developed an antigen quantification assay using ABC values. Antigen quantification as a clinical assay requires methods for quality control and for interlaboratory and inter-cytometer platform standardization. A single lot of Cytotrol™ Lyophilized Control Cells (Beckman Coulter) used for all studies. The variability in antigen quantification across 4 different instrument platforms in 2 separate laboratories was evaluated. The effect of the antibody clone utilized, importance of custom 1:1 molar ratio (fluorophore to protein, F/P) verses off-the-shelf antibodies, and QuantiBrite PE calibration verses linearity calibration combined with a single point scale transformation with CD4 as reference were determined. Use of single lot control cells allowed validation of reproducibility between flow cytometer platforms and laboratories and allowed assessment of different antibody lots, cocktail preparation, and different antibody clones. Off the shelf antibody preparations provide reproducible estimates of antigen density, however custom 1:1 unimolar antibody preparations should be utilized for definitive measurement of antigen expression.Geometric Mean fluorescent Intensity (GeoMFI) was not comparable across instruments and inter-laboratory. The use of CD4 as the reference marker can minimize variability in ABC values. Comparable antigen quantification is vital in managing patients receiving antigen-based immunotherapy. If this assay is to be utilized in a clinical setting, quality control methods have to be instituted to assure reproducibility and allow validation across laboratories. We have demonstrated that use of a lyophilized cell control is highly valuable in achieveing these goals.

Campylobacter species and genotype distribution in Finnish beef liver – Retail liver juice ideal for isolation and quantification

Campylobacteriosis, primarily caused by Campylobacter jejuni and C. coli, is the main bacterial zoonosis worldwide. While poultry is recognized as the main reservoir, bovines are considered another important reservoir for Campylobacter spp. found in human infections. In contrast to chicken, retail beef is seldom contaminated by Campylobacter species. However, beef liver is recognized to be frequently contaminated and has been linked to human infections via epidemiological investigations. Our aims were to evaluate the prevalence of Campylobacter spp. inside and on the surface of beef liver pieces at retail in Finland and to analyse the population in more detail using whole genome sequencing (WGS) to assess the public health relevance. A total of 90 retail beef livers were studied using both enrichment of the external peptone-saline rinse of the liver piece and direct culture from the inside after surface sterilization. Furthermore, 46 of the livers were also studied using direct culture of retail beef liver juice collected from the bottom of the consumer package to estimate the concentration of Campylobacter species. Overall, 44 (49 %) of the samples were positive for Campylobacter species, C. jejuni, C. fetus and C. lari being identified in 42 %, 8.9 % and 1.1 % of the samples, respectively. Direct culture of retail liver juice was a sensitive and convenient method for Campylobacter spp. detection, resulting in 48 % prevalence and a mean concentration of 49 cfu/ml (maximum 335 cfu/ml). Two samples (2.2 %), containing large hepatic ducts, were positive for C. jejuni internally, representing multilocus sequence typing (MLST) sequence type ST-19 and ST-21. WGS, core genome phylogeny and core genome MLST revealed that in most cases only one clearly distinct clone of clinically relevant C. jejuni or C. fetus was isolated from a single lot of samples. However, in some cases several distinct clones were identified simultaneously even from a single liver piece. In epidemiological investigations, it is thus highly advisable to genotype multiple isolates to capture the whole diversity of Campylobacter spp. from suspected food sources. Good kitchen hygiene, avoidance of cross-contamination and thorough cooking are important for limiting the transmission of campylobacteriosis.

Cellular Mechanisms Affecting Allogeneic CAR T Cell Expansion and Rejection in Large B-Cell Lymphoma

Allogeneic chimeric antigen receptor (CAR) T cell therapies hold promise to improve CAR T access. However, limited expansion and persistence due to rejection in some recipients may impact efficacy. Elucidation of cellular mechanisms governing allogeneic CAR T expansion and persistence may inform patient management and technology development. We hypothesized that cell-extrinsic and intrinsic mechanisms differentially regulate rejection and expansion of allogeneic CAR T cells in patients with large B-cell lymphoma. We identified 11 patients with relapsed/refractory large B-cell lymphoma treated on the ALPHA-2 phase 1 trial (NCT04416984) with ALLO-501A, a healthy donor-derived CD19 CAR T with T cell receptor (TCR) knockout. ALLO-501A uses Cellectis technologies. Despite treatment with a single ALLO-501A lot, we observed heterogeneity in CAR T expansion and clinical outcomes. Six patients had robust expansion (expanders), and five patients had relatively poor expansion (non-expanders) [median maximum CAR vector copy number 21,571 (range 10,296-104,784) versus 141 (range 55-502) copies/µg DNA, p < 0.01]. As with autologous therapies, expanders exhibited longer CAR T persistence and a higher frequency of clinical responses than non-expanders. To elucidate cell-extrinsic mechanisms involved in allogeneic CAR T rejection, we hypothesized that recipient-derived alloreactive T cells limit expansion in non-expanders. We adapted a mixed lymphocyte reaction assay to identify alloreactive CD4 + and CD8 + T cell clones from pre-lymphodepletion PBMC samples. Briefly, labeled recipient PBMCs were incubated with irradiated ALLO-501A cells from the treatment lot. Proliferating recipient CD4 + and CD8 + T cells were isolated and underwent TCR sequencing. Alloreactive clones were defined as those enriched in this sample compared to an unstimulated sample from the same patient (p < 0.05 by Fisher's test) and were tracked post-treatment. Two expanders and two non-expanders were selected for analysis. Non-expanders exhibited more robust in vitro T cell proliferation in response to ALLO-501A than expanders (85 vs. 40% proliferating; p = 0.03). Thus, more alloreactive clones were identified in non-expanders (median 762 CD4 + clones, 314 CD8 + clones) than expanders (median 313 CD4 + clones, 90 CD8 + clones). Non-expanders also had higher median alloreactive CD8 + T cell frequencies than expanders at days 7 and 14 following allogeneic CAR T infusion ( Figure A). A similar pattern was not observed for alloreactive CD4 + clones or non-alloreactive T cell clones, highlighting the potential role of recipient alloreactive CD8 + T cells in the rejection process and the need to control this process for the success of allogeneic CAR T. Next, to define the role of cell-intrinsic mechanisms in allogeneic CAR T expansion, we tracked clonal evolution of the CAR T repertoire through TCR sequencing of the infusion product (IP) and post-treatment PBMC samples in all patients. At day 14 (peak allogeneic CAR T expansion), all expanders had numerous IP-derived clones and substantial clonotype overlap with each other. In contrast, no clonotype overlap was observed among non-expanders ( Figure B). Furthermore, we identified the same high-frequency IP-derived clones in multiple expanders and their relative abundance reflected the original clonal hierarchy of the IP. This pattern differs from autologous therapies, where clonal hierarchy is not maintained after infusion (Sheih, et al. Nat. Comm. 2020), and suggests that cell-intrinsic mechanisms may influence allogeneic CAR T expansion potential. Single-cell RNA and proteomic studies to characterize these expanding clones are ongoing. In conclusion, we compared expansion kinetics across 11 allogeneic CAR T recipients to elucidate the cellular mechanisms governing allogeneic CAR T expansion and rejection. We revealed the impact of recipient alloreactive CD8 + T cells in allogeneic CAR T rejection and identified expansion of identical allogeneic CAR T clonotypes with retained clonal hierarchy in multiple patients. While patient factors such as tumor burden and target antigen expression likely play a role as well, our results suggest that strategies to eliminate alloreactive recipient cells and enhance allogeneic CAR T fitness through improved product design, manufacturing and lymphodepletion could improve allogeneic CAR T expansion, persistence and efficacy.

Preclinical Efficacy of a Novel CD8-Targeted Fusosome for In Vivo CAR T Cell Therapy

SG299 is a CD8-targeted fusosome; a self-inactivating (SIN) lentiviral vector (LVV) pseudotyped with a CD8-targeted fusogen, which carries a transgene encoding a CD19-directed chimeric antigen receptor (CAR). We evaluated the preclinical efficacy of a representative CD8/CD19CAR fusosome comparable to SG299 for its ability to control tumor growth in vivo in an NSG mouse model engrafted with human CD19 positive tumor cells and human peripheral blood mononuclear cells (PBMC). We compared direct intravenous (IV) administration and ex vivo exposure of fusosome to resting T cells followed by IV injection of vector-cell mixtures, as has been proposed for extracorporeal delivery (ECD) methods. We have also evaluated the performance of multiple lots of fusosomes to assess consistency in manufacturing and have evaluated donor-to-donor variability in five PBMC donors in this model. To measure anti-tumor efficacy, immunodeficient NSG mice were challenged IV with NALM6 tumors expressing firefly luciferase (5E5 cells/mouse) on day 4. For direct IV, PBMC (1E7 cells/mouse) were injected IV on day -1 followed by CD8/CD19CAR fusosome (1E7 IU/mouse) on day 0. For ECD, PBMC were incubated with CD8/CD19CAR fusosome for 1 hr and the cell and fusosome mix were injected IV on day 1. Tumor growth was monitored by bioluminescence imaging over a 28-day period. In addition, we have investigated both pharmacokinetics of fusosome induced CAR T cells and NALM6 tumors by flow cytometry. CD8/CD19CAR fusosome demonstrated statistically significant control of NALM6 tumors by both direct IV and ECD routes of delivery. Kinetic analysis of peripheral blood demonstrated tumor control and in vivo CART cell generation were dose dependent. The peak CAR T cell response was at D14. CARs detected at all timepoints were CD8+ specific; no CD4 + CAR T cells were observed. In addition, all mice that received fusosome demonstrated protection from morbidity and mortality as measured by reduced weight loss and improved survival compared to control or tumor only mice. Lot-to-lot performance variability of fusosomes was minimal; three lots of fusosome that had similar physical and functional attributes showed efficient and equivalent tumor control with multiple PBMC donors. Finally, an assessment of donor-to-donor variability across five PBMC donors using a single lot of fusosome showed donor-specific variations in tumor control as measured by BLI but demonstrated a consistent protection from morbidity and mortality. Collectively, these studies demonstrate preclinical efficacy of our CD8/CD19CAR candidate, lot consistency and verify our ability to generate CD19-directed CAR T cells in vivo using a specifically targeted platform.

A Study on Two-Warehouse Inventory Systems with Integrated Multi-Purpose Production Unit and Partitioned Rental Warehouse

A study of two warehouse inventory systems with a production unit is developed in this article with some constraints which are of practical applicability to optimize the total production cycle and its cost. A production unit evolves in three different states to retain its quality and prolong its lifetime: the state of producing items, the state of reworking the identified defective items, and the state of being idle. It processes the items up to a certain time point. The screening process starts immediately after a product comes out of the production unit. The classified non-defective items are first stored in own warehouse (OW), after filling to its maximum capacity, and the remaining items fill in the first block RW1 of the rental warehouse RW. All identified defective items are stored in the second block RW2 of RW. The holding cost of an item is higher in RW than OW. All defective items are sent to the production unit for re-do processes as a single lot immediately after the stop of the production and re-do items are stored in RW1 to satisfy the demand. The items in the RW1 are of higher priority in satisfying the demands after the stop of the production unit in producing new items as to deduce the total cost. Demand is assumed as both time and advertisement dependent and is encouraged once production starts. The deterioration rate differs in both warehouses. No backlog is entertained. The study is directed to achieve optimum total cycle cost towards the attainment of the optimum production time slot and the entire cycle of the system. We have arrived at explicit expressions for the total cost function of the entire production cycle. An analytic optimization process of the discriminant method is employed in the form of an algorithm to arrive at the optimum total cost. It provides a numerical illustration of a specific environment. The implications of the current research work are as follows. The optimum utility of production units in three different states in arriving at the optimum total cost is extensively studied with respect to deterioration, demand, and production rates. It also examined the influence of fluctuating deterioration, demand, and production parameters in arriving at optimum deterioration cost, holding cost, and total cycle cost, as they have important managerial insights. The effect of rental charges on the optimum total cost is examined as the system is used for multi-purpose storage.

Levels and Distribution of Salmonella in Naturally Contaminated Cashews

Raw materials associated with foodborne illness outbreaks are rarely available for evaluation. The levels and distribution of Salmonella were determined in naturally contaminated raw cashews linked to a salmonellosis outbreak associated with a fermented cashew cheese analog. Two unopened 22.7-kg boxes from a single lot of cashew kernel pieces were each divided into seven approximately equal units, 14 in total. Three 10-g subsamples per unit (n = 21) were evaluated for aerobic plate count (APC), coliform counts, and Escherichia coli counts, and 10 50-g subsamples per unit (n = 70) were enriched for the presence of Salmonella. Presumptive Salmonella-positive colonies were confirmed using CHROMagar Salmonella and real-time PCR (invA) and then serotyped using antigenic methods and genome sequencing prediction tools. APC and coliform counts ranged from 1.81 to 5.47 (mean 2.44 ± 0.63) log CFU/g and 0.60 to 5.20 (mean 1.74 ± 0.80) log CFU/g, respectively. Salmonella was recovered from four units in Box 1 and all seven units in Box 2. One of the 10 subsamples was positive in all but four of the positive units; one (Box 1) and three (Box 2) units had two positive subsamples. The level of Salmonella in the two boxes combined was 0.0023 most probable number/g (95% confidence interval [0.0014, 0.0038]). Salmonella Urbana was isolated from three of five positive subsamples in Box 1 and eight of 10 positive subsamples in Box 2. Salmonella Fresno and Vinohrady were unique to single subsamples from Box 1, and Salmonella Nima was isolated from two subsamples from Box 2. Of the four serovars recovered, Salmonella Urbana and Salmonella Vinohrady were in common with outbreak-associated clinical or product isolates. Understanding the distribution and concentration of Salmonella in naturally contaminated cashews provides important information for hazard analysis and risk assessments for soaked and fermented cashew products.

Chemical Constituents (Proximate, Minerals, Bioactive Compounds) of Pea [Pisum sativum (L.)] Shells

Background: Peas are preserved by freezing in pea processing industries because of seasonal limitations; there is a vast amount of pea peels as solid waste generated. Now days, the nutritional and techno-functional properties of by-products play an important role in the development of functional and enriched products in the food sector. The current study aim to analyze the nutrients content in pea shells. The result suggests pea shells can be used for value-addition in products and presence of antioxidant in pea shell powder indicates a better health effect if consumed. Methods: In this laboratory investigation during 2017-2018 peas were collected from market a single lot and analyzed for chemical constituents. In this connection, a fibrous coat of pea shells was separated, dried and prepared powder to perform nutritional analysis. Result: The study result indicates that pea shells have high nutritive value, e.g. protein (17.76%), total dietary fiber (21.04%), calcium (803.33 mg/100 g), iron (10.70 mg/100 g), potassium (1078.75 mg/100 g), magnesium (1029.55 mg/100 g) and bioactive compounds. Fresh pea shells have higher antioxidant activity (91%) compared to powder (86%). The total phenols, vitamin C and â-carotene strongly correlated with antioxidant activity. During the storage study, total phenols and antioxidant activity significantly decreased while on 90 days it was increased.

Second‐generation fully automated Elecsys cerebrospinal fluid immunoassays demonstrate high precision, reproducibility, and sample stability suitable for clinical use to aid Alzheimer’s disease diagnosis

AbstractBackgroundBeta‐amyloid(1–42) (Abeta42) and phosphorylated tau 181 (pTau) cerebrospinal fluid (CSF) biomarker analysis is recommended alongside cognitive evaluation to aid Alzheimer’s disease (AD) diagnosis. For clinical use, CSF biomarker assays should demonstrate high precision/reproducibility and sample stability. This study evaluated precision, reproducibility, and sample stability of second‐generation fully automated Elecsys® CSF immunoassays.MethodFrozen/pooled CSF and PreciControl samples were analyzed using Elecsys β‐Amyloid(1–42) CSF II/Phospho‐Tau (181P) CSF immunoassays on cobas e 601 (Roche Diagnostics International Ltd). pTau/Abeta42 ratios were calculated. Precision was evaluated at one site per Clinical and Laboratory Standards Institute (CLSI) EP02‐A3 (two runs/day in duplicate over 21 days; n=84). Reproducibility was determined per CLSI EP05‐A3 over five days using a single lot across three sites (site‐to‐site)/three lots at one site (lot‐to‐lot). Coefficients of variation (CVs)/standard deviations (SDs) for repeatability/intermediate precision/reproducibility were calculated using variance component analysis. Sample stability was determined from fresh CSF, collected per manufacturer's instructions. Baseline measurements were taken <6 hours after lumbar puncture and follow‐up measurements (from stored aliquots) at defined intervals for ≤8 days at room temperature (RT; 25°C; n=13), ≤15 days at 6°C (n=14), and ≤13 weeks at ‐20°C (n=25). Storage stability was determined from percent recoveries/ordinary linear regression.ResultRepeatability CVs were <2.4% and <1.9% for Abeta42 and pTau immunoassays, respectively, and <2.4% for pTau/Abeta42 ratio. Intermediate precision CVs were <3.3% and <3.7% for Abeta42 and pTau immunoassays, respectively, and <3.9% for pTau/Abeta42 ratio. Site‐to‐site reproducibility CVs were <5.1% and <3.6% for Abeta42 and pTau immunoassays, respectively, and <6.5% for pTau/Abeta42 ratio (Table 1). For the Abeta42 immunoassay, lot‐to‐lot reproducibility CVs were <5.4%, except for one sample pool (mean concentration: 322 pg/mL; SD: 34.1 pg/mL; Table 2). Lot‐to‐lot reproducibility CVs were <3.6% for the pTau immunoassay and <6.0% for pTau/Abeta42 ratio. At maximum sample storage duration (8 days at RT/15 days at 2−8°C), median recoveries of 95%/98% were observed for Abeta42 and 97%/99% for pTau immunoassays. Stability assessment at ‐20°C is ongoing.ConclusionThe Elecsys immunoassays demonstrated high precision/reproducibility, supporting clinical use in AD diagnosis. Conservative recommendations for sample storage are 5 days at RT/14 days at 2−8°C.

Effects of cushion box and closed let-down ladder usage on impact damage to corn kernel during handling.

The main purpose of this study was to evaluate the effects of the cushion box and closed let-down ladder usage in minimizing mechanical damage to corn kernels during free fall. Kernels from a single lot of cultivar KSC 705 were evaluated for percentage of breakage using three drop methods (free fall, with cushion box, and with closed let-down ladder) at five different moisture contents (10%, 15%, 20%, 25%, and 30%), and three drop heights (5, 10, and 15 m). The results showed that the drop methods had a significant effect on the breakage sensibility of kernels. Sample kernels dropped without a ladder (free fall) had a significantly higher average percentage breakage of 13.80%. In the use of the cushion box, the average kernel breakage was calculated to be 11.41%, which was decreased by about 17% more than the free fall. Sample kernels dropped with the closed let-down ladder had a lower average breakage of 7.26%, which showed that the closed let-down ladder significantly helped to reduce mechanical damage to corn kernels by about 47% comparing free fall and by about 37% than the use of the cushion box. The amounts of kernel damage increased significantly with increasing drop height and decreasing moisture content, but the use of the cushion box and closed let-down ladder systems somewhat reduced the adverse effect of the above factors. To minimize mechanical damage to kernels as they fall into the bin, a grain let-down ladder should be installed in the bin so that it can receive kernels from the filling spout with minimum damage. Empirical models were developed for the dependency of damage to corn kernels due to the impact caused by free fall on the drop height and moisture content at different drop methods.