Preservation Solution Research Articles

Context Privacy Preservation for User Validation by Wireless Sensors in the Industrial Metaverse Access System

The Industrial Metaverse provides unparalleled prospects for increasing productivity and efficiency across multiple sectors. As wireless sensor networks play an important role in data collection and transmission within this ecosystem, preserving context privacy becomes critical to protecting sensitive information. This paper investigates the issue of context privacy preservation for user validation via AccesSensor in the Industrial Metaverse and presents a technological method to address it. We explore the need for context privacy, look at existing privacy preservation solutions, and propose novel user validation methods that are customized to the Industrial Metaverse’s access system. This method is evaluated on time-based efficiency, privacy method and bandwidth utilization. Our method performs better as compared to the DPSensor. Our research seeks to provide insights and recommendations for developing strong privacy protection methods in wireless sensor networks that operate within the Industrial Metaverse ecosystem.

Assessment of Mitochondrial Respiration During Hypothermic Storage of Liver Biopsies Following Normothermic Machine Perfusion

Organ quality can be assessed prior to transplantation, during normothermic machine perfusion (NMP) of the liver. Evaluation of mitochondrial function by high-resolution respirometry (HRR) may serve as a viability assessment concept in this setting. Freshly collected tissue is considered as optimal sample for HRR, but due to technical and personnel requirements, more flexible and schedulable measurements are needed. However, the impact of cold storage following NMP before processing biopsy samples for mitochondrial analysis remains unknown. We aimed at establishing an appropriate storage protocol of liver biopsies for HRR. Wedge biopsies of 5 human livers during NMP were obtained and assessed by HRR. Analysis was performed after 0, 4, 8, and 12 h of hypothermic storage (HTS) in HTK organ preservation solution at 4°C. With HTS up to 4 h, mitochondrial performance did not decrease in HTS samples compared with 0 h (OXPHOS, 44.62 [34.75–60.15] pmol·s−1·mg wet mass−1 vs. 43.73 [40.69–57.71], median [IQR], p > 0.999). However, at HTS beyond 4 h, mitochondrial respiration decreased. We conclude that HTS can be safely applied for extending the biopsy measurement window for up to 4 h to determine organ quality, but also that human liver respiration degrades beyond 4 h HTS following NMP.

Innovative Agrowaste Banana Peel Extract-Based Magnetic Iron Oxide Nanoparticles for Eco-Friendly Oxidative Shield and Freshness Fortification

This study presents the synthesis of agro-waste banana peel extract-based magnetic iron oxide nanoparticles (BPEx-MIONPs), emphasizing antioxidant capacity and food preservation. Using iron (III) chloride hexahydrate (FeCl3 · 6 H2O) as a precursor and a reducing agent from agro-waste peel extract, a precisely controlled process yielded BPEx-MIONPs. Characterization involved X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and Fourier-transform infrared spectroscopy (FTIR). XRD revealed tetragonal Fe2O3, cubic magnetite structure, and monoclinic FexOy-NPs with an average size of 14.8 nm. TEM and SEM revealed diverse morphologies. TEM displayed both spherical and elongated nanoparticles, with some appearing as thin fibrils. In contrast, SEM images depicted an array primarily consisting of spherical nanoparticles, resembling coral reef formations. FTIR confirmed Fe–O bonds (1000 –400 cm-1). The antioxidant assessment showed robust DPPH free radical scavenging; BPEx achieved 100% inhibition at 18 min, and BPEx-MIONPs had an IC50 of ~ 136 µg/mL. BPEx-MIONPs, stabilized with banana-based bioplastic, effectively preserved grapes, reducing weight loss to 6.2% on day 3, compared to the control (19.0%). This pioneering study combines banana peel antioxidants with magnetic iron oxide nanoparticles, providing sustainable solutions for food preservation and nano-packaging. Ongoing research aims to refine conditions and explore broader applications of BPEx-MIONPs.Graphical

Preservation Solutions for Cuc Phuong National Park From a Tourism Perspective

Purpose: the purpose of this paper is to present solutions to preserve cuc phuong national park from tourism view Methods: The study employs qualitative methods including synthesis, inductive and explanatory methods. The overall purpose of this study is to obtain the understanding of Cuc Phuong National Park residents’ perceptions and satisfaction toward tourism impacts as well as their attitudes for tourism development. Besides that, this study also points out the main issues of current ecotourism strategies in Cuc Phuong National Park. Based on the results, ecotourism strategies were proposed to response to ecotourism criteria. The interview questions and questionnaire were developed in English To gain an in-depth understanding of local householders’ perceptions on ecotourism and the current ecotourism strategies in Cuc Phuong National Park. Results and Discussion: to improve the efficiency of conservation activity, Cuc Phuong National Park should give more power for local people in implementing and managing activities. Besides that, the law enforcement about illegal activities must to enhance in order to catch of the current context. Based on the practical observation of researcher, the current law is too weak to prevent the illegal activities. Therefore, stronger law policies are very necessary to promulgate soon and implement in strict and opened ways. Originality/Value: the results of study and the author’s proposed strategies will help Cuc Phuong National Park improve tourism activities in a sustainable way, harmony among benefits, especially focus on community and conservation. Besides that, it also helps the local government, tour-operators and tourism promoters be aware of real concerns, issues of current tourism activities in Cuc Phuong National Park and then have appropriate program designs and actions to be effective, satisfying community benefits and minimizing the problems for the natural environment.

An Integrated Testing Strategy (ITS) to assess the environmental compatibility of wood protection techniques

To quantify the possible impact of different wood protection techniques on the aquatic environment, we applied a tiered Integrated Testing Strategy (ITS) on leachates obtained from untreated (UTW) Norway spruce (Picea abies), specimens treated with a copper-ethanolamine-based preservative solution, complying with the Use Class 3 (UC3), and specimens thermally modified (TM). Different maturation times in water were tested to verify whether toxicant leaching is time-dependent. Tier I tests, addressing acute effects on Aliivibrio fischeri, Raphidocelis subcapitata, and Daphnia magna, evidenced that TM toxicity was comparable or even lower than in UTW. Conversely, UC3 significantly affected all species compared to UTW, also after 30 days of maturation in water, and was not considered an environmentally acceptable wood preservation solution. Tier II (effects on early-life stages of Lymnea auricularia) and III (chronic effects on D. magna and L. auricularia) performed on UTW and TM confirmed the latter as an environmentally acceptable treatment, with increasing maturation times resulting in decreased adverse effects. The ITS allowed for rapid and reliable identification of potentially harmful effects due to preservation treatments, addressed the choice for a less impacting solution, and can be effective for manufacturers in identifying more environmentally friendly solutions while developing their products.

Optimization of coagulation-flocculation process for wastewater treatment from vegetable oil refineries using chitosan as a natural flocculant

Vegetable oil refinery wastewater (VORW) is a significant source of refractory pollutants necessitating efficient treatment prior to discharge. This study investigates the treatment of VORW via coagulation-flocculation using ferric chloride (FeCl3) as a coagulant and chitosan as a natural flocculant. Central Composite Design (CCD) is employed to optimize the treatment process and assess the interplay of experimental factors. The study evaluates turbidity, COD, and polyphenols as responses, with pH, FeCl3 concentration, chitosan dosage, and agitation time as independent factors.The results showed that the optimal conditions identified include pH 6, FeCl3 dosage of 1.6 g/L, chitosan dosage of 13.4 mg/L, and agitation time of 26 min, resulting in 100 % turbidity removal, 86 % COD reduction, and 90 % polyphenol removal. The analysis of variance indicated that the established models were significant and that they are characterized by a good fit (R2 in the order of 0.95, 0.96, and 0.96 for turbidity, COD, and polyphenols, respectively).These findings highlight the efficacy and sustainability of the coagulation-flocculation process with chitosan, offering a practical, rapid, and cost-effective solution for VORW treatment and environmental preservation.

Minimally Invasive Management of Uterine Fibroids - Role of Transvaginal Radiofrequency Ablation

Minimally invasive approaches in management of common gynecological pathologies such as uterine fibroids are of outmost importance to offer fertility preservation and cost-effective solutions. Transvaginal radiofrequency ablation in day surgery setting offers an efficient volume reduction solution with improvement of fibroid related symptoms in patients undergoing accurate pre-operative selection.

Toward an optimal cadaveric brain model for neurosurgical education: assessment of preservation, parenchyma, vascular injection, and imaging.

We assessed types of cadaveric head and brain tissue specimen preparations that are used in a high throughput neurosurgical research laboratory to determine optimal preparation methods for neurosurgical anatomical research, education, and training. Cadaveric specimens (N = 112) prepared using different preservation and vascular injection methods were imaged, dissected, and graded by 11 neurosurgeons using a 21-point scale. We assessed the quality of tissue and preservation in both the anterior and posterior circulations. Tissue quality was evaluated using a 9-point magnetic resonance imaging (MRI) scale. Formalin-fixed specimens yielded the highest scores for assessment (mean ± SD [17.0 ± 2.8]) vs. formalin-flushed (17.0 ± 3.6) and MRI (6.9 ± 2.0). Cadaver assessment and MRI scores were positively correlated (P < 0.001, R2 0.60). Analysis showed significant associations between cadaver assessment scores and specific variables: nonformalin fixation (β = -3.3), preservation within ≤72h of death (β = 1.8), and MRI quality score (β = 0.7). Formalin-fixed specimens exhibited greater hardness than formalin-flushed and nonformalin-fixed specimens (P ≤ 0.006). Neurosurgeons preferred formalin-flushed specimens injected with colored latex. For better-quality specimens for neurosurgical education and training, formalin preservation within ≤72h of death was preferable, as was injection with colored latex. Formalin-flushed specimens more closely resembled live brain parenchyma. Assessment scores were lower for preparation techniques performed > 72h postmortem and for nonformalin preservation solutions. The positive correlation between cadaver assessment scores and our novel MRI score indicates that donation organizations and institutional buyers should incorporate MRI as a screening tool for the selection of high-quality specimens.

Steatotic Donor Transplant Livers: Preservation Strategies to Mitigate against Ischaemia-Reperfusion Injury.

Liver transplantation (LT) is the only definitive treatment for end-stage liver disease, yet the UK has seen a 400% increase in liver disease-related deaths since 1970, constrained further by a critical shortage of donor organs. This shortfall has necessitated the use of extended criteria donor organs, including those with evidence of steatosis. The impact of hepatic steatosis (HS) on graft viability remains a concern, particularly for donor livers with moderate to severe steatosis which are highly sensitive to the process of ischaemia-reperfusion injury (IRI) and static cold storage (SCS) leading to poor post-transplantation outcomes. This review explores the pathophysiological predisposition of steatotic livers to IRI, the limitations of SCS, and alternative preservation strategies, including novel organ preservation solutions (OPS) and normothermic machine perfusion (NMP), to mitigate IRI and improve outcomes for steatotic donor livers. By addressing these challenges, the liver transplant community can enhance the utilisation of steatotic donor livers which is crucial in the context of the global obesity crisis and the growing need to expand the donor pool.

Hypothermic machine perfusion of a donor kidney using an experimental dextran-40-based preservation solution and orthotopic transplantation (experimental study)

Objective: to evaluate the efficacy of hypothermic machine perfusion (HMP) of a donor kidney obtained from a non-heartbeating (NHB) donor, using an experimental dextran-40-based preservation solution, in subsequent orthotopic transplantation in a rabbit model.Materials and methods. Twenty grey giant rabbits weighing 2,500–3,100 g, divided into donors (n = 10) and recipients (n = 10), were used in the study. After obtaining kidney from an NHB donor, ex vivo HMP of the left donor kidney using a dextran-40-based preservation solution was performed and peripheral vascular resistance (PVR) parameters were measured. This was followed by bilateral nephrectomy and orthotopic transplantation. The follow-up period was 12 days. Creatinine levels, urea levels, and glomerular filtration rate (GFR) were measured during follow-up.Results. During ex vivo HMP of donor kidneys from NHBs, PVR dropped progressively from 1.90 ± 0.27 mmHg/mL/min to 0.72 ± 0.09 mmHg/mL/min at p &lt; 0.001. In the early post-transplant period (during the first 2 days after implantation), creatinine and urea levels were moderately elevated compared to normal. Creatinine and urea levels were 91.07 ± 11.49 μmol/L at p &lt; 0.011 and 9.09 ± 1.06 mmol/L at p &lt; 0.009 on day 2, respectively, but by day 12, they reverted to physiologic values, which were 77.17 ± 10.19 μmol/L at p &lt; 0.019 and 4.88 ± 0.54 mmol/L at p &lt; 0.022, respectively. These findings were correlated with GFR values, which ranged from 26.29 to 26.74 mL/min/1.72 m2 in mean values over the course of a 12-day follow-up period.Conclusion. Ex vivo HMP using dextran-40-based preservation solution has a positive effect on the kidney at 30 minutes of warm ischemia following asystole and achieves satisfactory graft function over 12 days of follow-up.

Liraglutide Protects Pancreatic Islet From Ischemic Injury by Reducing Oxidative Stress and Activating Akt Signaling During Cold Preservation to Improve Islet Transplantation Outcomes.

Islet transplantation is a promising therapy for patients with type 1 diabetes. However, ischemic injury to the donor islets during cold preservation leads to reduced islet quality and compromises transplant outcome. Several studies imply that liraglutide, a glucagon-like peptide-1 receptor agonist, has a positive effect on promoting islet survival, but its impact on islet cold-ischemic injury remains unexplored. Therefore, the aim of this study was to investigate whether liraglutide can improve islet transplantation efficacy by inhibiting cold-ischemic injury and to explore the underlying mechanisms. Liraglutide was applied in a mouse pancreas preservation model and a human islets cold-preservation model, and islet viability, function, oxidative stress levels were evaluated. Furthermore, islet transplantation was performed in a syngeneic mouse model and a human-to-nude mouse islet xenotransplantation model. The supplementation of liraglutide in preservation solution improved islet viability, function, and reduced cell apoptosis. Liraglutide inhibited the oxidative stress of cold-preserved pancreas or islets through upregulating the antioxidant enzyme glutathione levels, inhibiting reactive oxygen species accumulation, and maintaining the mitochondrial membrane integrity, which is associated with the activation of Akt signaling. Furthermore, the addition of liraglutide during cold preservation of donor pancreas or donor islets significantly improved the subsequent transplant outcomes in both syngeneic mouse islet transplantation model and human-to-nude mouse islet xenotransplantation model. Liraglutide protects islets from cold ischemia-related oxidative stress during preservation and hence improved islet transplantation outcomes, and this protective effect of liraglutide in islets is associated with the activation of Akt signaling.

Cell-specific Extracellular Vesicles and Their miRNA Cargo Released Into the Organ Preservation Solution During Cold Ischemia Storage as Biomarkers for Liver Transplant Outcomes.

Liver transplantation (LT) is crucial for end-stage liver disease patients, but organ shortages persist. Donation after circulatory death (DCD) aims to broaden the donor pool but presents challenges. Complications like acute rejection, hepatic artery thrombosis, and biliary issues still impact posttransplant prognosis. Biomarkers, including extracellular vesicles (EVs) and microRNAs (miRNAs), show promise in understanding and monitoring posttransplant events. This study explores the role of EVs and their miRNA cargo in LT, including their potential as diagnostic tools. EVs from intrahepatic end-ischemic organ preservation solution (eiOPS) in 79 donated livers were detected using different techniques (nanosight tracking analysis, transmission electron microscopy, and flow cytometry). EV-derived miRNAs were identified by quantitative real time-polymerase chain reaction. Bioinformatics analysis was performed using the R platform. Different-sized and origin-specific EVs were found in eiOPS, with significantly higher concentrations in DCD compared with donation after brain death organs. Additionally, several EV-associated miRNAs, including let-7d-5p , miR-28-5p , miR-200a-3p , miR-200b-3p , miR-200c-3p , and miR-429 , were overexpressed in DCD-derived eiOPS. These miRNAs also exhibited differential expression patterns in liver tissue biopsies. Pathway analysis revealed enrichment in signaling pathways involved in extracellular matrix organization and various cellular processes. Moreover, specific EVs and miRNAs correlated with clinical outcomes, including survival and early allograft dysfunction. A predictive model combining biomarkers and clinical variables showed promise in acute rejection detection after LT. These findings provide new insights into the use of EVs and miRNAs as biomarkers and their possible influence on posttransplantation outcomes, potentially contributing to improved diagnostic approaches and personalized treatment strategies in LT.

Advancements in bamboo preservative solutions for sustainable construction in Brazil

Advancements in bamboo preservative solutions for sustainable construction in Brazil

Persistence of SARS-CoV-2 colonization and high expression of inflammatory factors in cardiac tissue 6 months after COVID-19 recovery: a prospective cohort study.

The presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in myocardial autopsy tissues has been observed in certain individuals with coronavirus disease 2019 (COVID-19). However, the duration of cardiac involvement remains uncertain among recovered COVID-19 patients. Our study aims to evaluate the long-term persistence of SARS-CoV-2 within cardiac tissue. We prospectively and consecutively evaluated the patients undergoing mitral valve replacement (MVR) and left atrial (LA) volume reduction surgery from May 25 to June 10, 2023 at our center, who had been approximately 6 months of recovery after Omicron wave. Patients tested positive for SARS-CoV-2 upon admission were excluded. The surgical LA tissue was collected in RNA preservation solution and stored at -80 ℃ immediately. Then SARS-CoV-2, interleukin-6 (IL-6) and interleukin-1β (IL-1β) RNA expression in LA tissues were assessed through thrice-repeated reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analyses. Categorical variables were assessed using the Chi-square or Fisher's exact tests, and continuous variables was analyzed using the Mann-Whitney U test. Nine of 41 patients were enrolled, all of whom tested negative for SARS-CoV-2 upon admission (two antigen and PCR tests). In four of nine patients, SARS-CoV-2 RNA was detected in their LA tissue, indicating viral colonization. Among the four positive cases, the IL-6 and IL-1β relative expression levels in the LA tissue of one patient were increased approximately 55- and 110-fold, respectively, compared to those of SARS-CoV-2 (-) patients. Increased expression of IL-6 and IL-1β were observed in the myocardium of this patient. Another patient demonstrated a remarkable 7-fold increase in both IL-6 and IL-1β expression, surpassing that of SARS-CoV-2 (-) patients. Additionally, no other cardiac inflammation-related diseases or conditions were presented in these two patients. The IL-6 and IL-1β expression levels of the remaining two patients were not significantly different from those of SARS-CoV-2 (-) patients. The relative expression levels of IL-6 and IL-1β in cardiac tissues of all SARS-CoV-2 (-) patients were relatively low. Interestingly, despite abnormally elevated levels of IL-6 and IL-1β within their cardiac tissue, two patients did not show a significant increase in serum IL-6 and IL-1β levels when compared to other patients. Our research suggests that certain COVID-19-recovered patients have persistent colonization of SARS-CoV-2 in their cardiac tissue, accompanied by a local increase in inflammatory factors.

(680) - Itaconate Supplementation in Preservation Solution Leads to Lung Function Improvement After Extended Hypothermic Preservation

(680) - Itaconate Supplementation in Preservation Solution Leads to Lung Function Improvement After Extended Hypothermic Preservation

(43) - A Newly Produced Polymerized Bovine Hemoglobin and Hemoglobin-Based Heart Preservation Solution Prolonged Preservation Duration of Donor Heart in Cold Static Storage and Reduced Myocardial Injury After Allograft Transplantation

(43) - A Newly Produced Polymerized Bovine Hemoglobin and Hemoglobin-Based Heart Preservation Solution Prolonged Preservation Duration of Donor Heart in Cold Static Storage and Reduced Myocardial Injury After Allograft Transplantation

Multi-Walled Carbon Nanotubes Improve the Development of Chrysanthemum × morifolium (Ramat.) Hemsl. ‘Jinba’ Inflorescences

The application of cut flower preservation technology can significantly enhance both the ornamental and economic value of fresh-cut flowers. Research on vase solutions has become a concentrated area in current studies on cut flower preservation. Multi-walled carbon nanotubes (MWCNTs), as a type of carbon nanomaterial with bactericidal and membrane-penetrating properties, can be used as a component in vase solutions. This supplementation of energy substances aims to improve antioxidant enzyme activity, thereby enhancing the postharvest quality of cut chrysanthemums. In this study, deionized water and a standard preservative solution were employed as control groups to compare the effects of MWCNTs applied at different concentrations, combined with common preservatives such as sugar and 8-hydroxyquinoline, on the postharvest flowering and preservation of Chrysanthemum × morifolium ‘Jinba’. By observing the distribution of MWCNTs in the tissues surrounding the cut and changes in water content, carbon sources, osmoregulatory substance levels, and the expression of relevant key genes, a formulation with excellent postharvest treatment effects was identified. Preliminary investigations into its action and mechanism were also conducted. The results indicated that the combined treatment with 5 mg L−1 MWCNTs, 30 g L−1 sucrose, and 0.2 g L−1 8-hydroxyquinoline effectively promoted water and sugar uptake in chrysanthemum flowers, accelerating bud opening, maintaining larger inflorescence diameter, and extending the vase life. Ultimately, this enhanced the ornamental value of cut chrysanthemums. These research findings provide theoretical and experimental foundations for the application of multi-walled carbon nanotubes as auxiliary additives to improve the ornamental quality of cut flowers.

Experimental study on the preservation of amputated limb by low-temperature HTK preservation solution arterial perfusion

Objective: To evaluate the effect of HTK (Histidine-Tryptophan-Ketoglutarate solution) infusion on the protection and replantation of amputated limb. Methods: In experiment 1, the amputated limbs of male New Zealand white rabbits were preserved by different preservation methods, including low-temperature HTK solution perfusion group, low-temperature normal saline perfusion group, low-temperature preservation group and non-perfusion group at normal temperature. The homogenates of muscle tissue at different time points were collected for biochemical detection. H&amp;E (hematoxylin and eosin) staining was performed on muscle tissue. Bax, Bcl-2 protein immunohistochemical staining and Tunel method were used to detect apoptosis in muscle, nerve, and vascular tissues. The ultrastructure of cells was observed by transmission electron microscope. In experiment 2, the amputated limbs of rabbits were treated with HTK perfusion and cryopreservation without perfusion, respectively. The survival of rabbits and their limbs were observed, and detected by H&amp;E, TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) and scanning electron microscopy. Statistical analysis was performed using SPSS 22 (SPSS Inc., Chicago, IL, USA). χ2 test, student’s t-test, and ANOVA (Analysis of Variance) were used to compare the categorical and continuous variables. Results: Experiment 1: With the extension of disconnection time, there were statistical differences between the biochemical indexes of the HTK liquid perfusion group and those of the non-low temperature non-perfusion group, while some indexes of the other groups were different. Immunohistochemical staining of tissues (muscle, nerve, and blood vessel) in each group showed that the number of cells stained by anti-apoptotic protein BCL-2 in each group significantly increased with the extension of time, while the number of cells stained by pro-apoptotic protein BAX2 in each group significantly decreased. Tunel test showed that compared with other treatment groups, the apoptosis rate of the HTK liquid group was significantly decreased. Experiment 2: The survival rate of the experimental group was 60%, and that of the control group was only 30%. H&amp;E staining, the TUNEL method and scanning electron microscopy suggested that HTK infusion could reduce ischemia-reperfusion injury in muscle tissue. Conclusion: 1. Perfusion preservation of severed limbs at 4 ℃ can effectively reduce tissue damage caused by ischemia and hypoxia, and the effect is stronger than direct preservation. 2. HTK solution and normal saline were used for one-time perfusion of the severed limbs of rabbits, and the preservation effect of HTK solution was the best. 3. HTK solution was used for one-time perfusion of severed hind limbs of rabbits, which was refrigerated at 4 ℃ and then replanted (ischemia time was 3–5 h). Compared with the replanted limbs after cold storage alone, the survival rate was higher, and the postoperative ischemia and hypoxia injury of surviving limbs were less.

Analysis of gene and protein expression in the endometrium for validation of an ex vivo model of the equine uterus using PCR, digital and visual histopathology

In the past, most research in equine reproduction has been performed in vivo but the use of in vitro and ex vivo models has recently increased. This study aimed to evaluate the functional stability of an ex vivo hemoperfused model for equine uteri with molecular characterization of marker genes and their proteins. In addition, the study validated the respective protein expression and the aptness of the software QuPath for identifying and scoring immunohistochemically stained equine endometrium. After collection, uteri (n = 12) were flushed with preservation solution, transported to the laboratory on ice, and perfused with autologous blood for 6 h. Cycle stage was determined by examination of the ovaries for presence of Graafian follicles or corpora lutea and analysis of plasma progesterone concentration (estrus: n = 4; diestrus: n = 4; anestrus: n = 4). Samples were obtained directly after slaughter, after transportation, and during perfusion (240, 300, 360 min). mRNA expression levels of progesterone (PGR), estrogen (ESR1) and oxytocin (OXTR) receptor as well as of MKI67 (marker of cell growth) and CASP3 (marker of apoptosis) were analyzed by RT-qPCR, and correlation to protein abundance was validated by immunohistochemical staining. Endometrial samples were analyzed by visual and computer-assisted evaluation of stained antigens via QuPath. For PGR, effects of the perfusion and cycle stage on expression were found (P < 0.05), while ESR1 was affected only by cycle stage (P < 0.05) and OXTR was unaffected by perfusion and cycle stage. MKI67 was lower after 360 min of perfusion as compared to samples collected before perfusion (P < 0.05). For CASP3, differences in gene expression were found after transport and samples taken after 240 min (P < 0.05). Immunohistochemical staining revealed effects of perfusion on stromal and glandular cells for steroid hormone receptors, but not for Ki-67 and active Caspase 3. OXTR was visualized in all layers of the endometrium and was unaffected by perfusion. Comparison of QuPath and visual analysis resulted in similar results. For most cell types and stained antigens, the correlation coefficient was r > 0.5. In conclusion, the isolated hemoperfused model of the equine uterus was successfully validated at the molecular level, demonstrating stability of key marker gene expression. The utility of computer-assisted immunohistochemical analysis of equine endometrial samples was also confirmed.

Abstract 335: BD®OMICS-Guard Sample Preservation Buffer preserves mRNA and cell surface epitopes for single-cell sequencing applications

Abstract Multiomics has emerged as a powerful tool to derive insights on the transcriptomic and proteomic data of human cancer samples and preclinical models of blood and tumor malignancies. Due to the sensitivity of sequencing techniques, sample quality and stability can potentially interfere with results. BD® OMICS-Guard Sample Preservation Buffer (SPB), a novel preservation solution, allows cells and bulk tissues to be stored for up to 72h at 4 °C, while reliably capturing the true biology of cells. This reagent gently preserves mRNA and protein integrity without the traditional cross-linking and harsh fixatives, increasing flexibility and expanding options for study designs. Here, we showcase the preservation properties ofthis reagent for multiomic readouts in human cells and mouse tissue across three different timepoints.BD® OMICS-Guard SPB’s preservation of mRNA from human PBMC and bulk mouse spleen tissues wasevidenced by WTA for differentially expressed genes in samples preserved for 24, 48 and 72h comparedto fresh/unpreserved controls. Protein epitope preservation was demonstrated via CITE-Seq assay withthe BD® AbSeq Human Immune Discovery Panel (human PBMC) or a 30-plex panel of anti-mouse BD®AbSeq Antibody-Conjugated Oligonucleotides (mouse splenocytes) and confirmed with multicolor flowcytometry. For both sample types, high correlation R2 values (&amp;gt;0.87) were calculated for the preservation time points versus fresh controls. Concomitant flow cytometry analysis of major cell types and surface protein expression was consistent with sequencing data for all samples. Specificity and sensitivity of major cell type markers and cell surface proteins across cell types and time is visualized in the BD®AbSeq Antibody-generated heatmap for each sample. Additionally, BD® OMICS-Guard SPB maintains mRNA integrity in targeted gene assays and VDJ full-length assays, suggesting robust preservation of select transcripts readily interrogated in single-cell profiling. Batch effect across time and technical replicates for both sample types was limited with the BD Rhapsody™ HT Xpress System and delineated by donor/sample specific tSNEs across time. We demonstrate that the BD® OMICS-Guard Sample Preservation Buffer effectively preserves both mRNA and surface proteins for single-cell experiments. BD® OMICS-Guard SPB allows the convenient preservation of samples such as tumor biopsies to be collected and stored for up to 72h at 4 °C while maintaining mRNA and protein integrity, allowing the flexibility for precious samples to be potentially shipped to a central sequencing site to ensure standardized sequencing workflows. For Research Use Only. Not for use in diagnostic or therapeutic procedures.BD and BD Rhapsody are trademarks of Becton, Dickinson and Company or its affiliates. © 2023 BD. All rights reserved. Citation Format: Samuel X. Shi, Cynthia Sakofsky, Zhiqi Zhang, Samatha Vadrevu, Xiaoshan Shi, Hye-Won Song, Aruna Ayer. BD®OMICS-Guard Sample Preservation Buffer preserves mRNA and cell surface epitopes for single-cell sequencing applications [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 335.