Tissue Retention Research Articles

The Mechanisms of Cadmium Toxicity in Living Organisms

Cadmium (Cd) is a toxic metal primarily found as a by-product of zinc production. Cd was a proven carcinogen, and exposure to this metal has been linked to various adverse health effects, which were first reported in the mid-19th century and thoroughly investigated by the 20th century. The toxicokinetics and dynamics of Cd reveal its propensity for long biological retention and predominant storage in soft tissues. Until the 1950s, Cd pollution was caused by industrial activities, whereas nowadays, the main source is phosphate fertilizers, which strongly contaminate soil and water and affect human health and ecosystems. Cd enters the human body mainly through ingestion and inhalation, with food and tobacco smoke being the primary sources. It accumulates in various organs, particularly the kidney and liver, and is known to cause severe health problems, including renal dysfunction, bone diseases, cardiovascular problems, and many others. On a cellular level, Cd disrupts numerous biological processes, inducing oxidative stress generation and DNA damage. This comprehensive review explores Cd pollution, accumulation, distribution, and biological impacts on bacteria, fungi, edible mushrooms, plants, animals, and humans on a molecular level. Molecular aspects of carcinogenesis, apoptosis, autophagy, specific gene expression, stress protein synthesis, and ROS formation caused by Cd were discussed as well. This paper also summarizes how Cd is removed from contaminated environments and the human body.

Enhanced model antigen retention in tissue through topical high-frequency ultrasound treatment

Enhanced model antigen retention in tissue through topical high-frequency ultrasound treatment

Magnetic soft microrobots for erectile dysfunction therapy

Erectile dysfunction (ED) is a major threat to male fertility and quality of life, and mesenchymal stromal cells (MSCs) are a promising therapeutic option. However, therapeutic outcomes are compromised by low MSC retention and survival rates in corpus cavernosum tissue. Here, we developed an innovative magnetic soft microrobot comprising an ultrasoft hydrogel microsphere embedded with a magnetic nanoparticle chain for MSC delivery. This design also features phenylboronic acid groups for scavenging reactive oxygen species (ROS). With a Young's modulus of less than 1 kPa, the ultrasoft microrobot adapts its shape within narrow blood vessels, ensuring a uniform distribution of MSCs within the corpus cavernosum. Our findings showed that compared with traditional MSC injections, the MSC delivery microrobot (MSC-Rob) significantly enhanced MSC retention and survival. In both rat and beagle ED models, MSC-Rob treatment accelerated the repair of corpus cavernosum tissue and restored erectile function. Single-cell RNA sequencing (scRNA-seq) revealed that MSC-Rob treatment facilitates nerve and blood vessel regeneration in the corpus cavernosum by increasing the presence of regenerative macrophages. Overall, our MSC-Rob not only advances the clinical application of MSCs for ED therapy but also broadens the scope of microrobots for other cell therapies.

Investigation of the usefulness of collagen peptide in intramyocardial injection of human iPS cell-derived cardiomyocytes

Abstract Background Injecting human induced pluripotent stem (iPS) cell-derived cardiomyocytes directly into the heart muscle holds promise as a treatment for heart failure. Achieving better engraftment is crucial for maximizing therapeutic effectiveness. Collagen peptide (CP) is a form of hydrolyzed gelatin known for its resistance to solidification. By adjusting its concentration, the viscosity of CP can be easily managed, potentially influencing the retention and engraftment efficiency of transplanted cells. Purpose We investigated the impact of different CP concentrations as a solvent for intramyocardial injection of human iPS cell-derived cardiomyocytes. Methods Initially, we examined the leakage of CP solution from the injection site using chicken meat and ex vivo rat heart. Subsequently, an in vivo experiment was conducted using beating rat myocardium injected with CP solution and Indian ink to assess CP diffusion in dynamic heart tissue. Finally, we performed intramyocardial injections of human iPS cell-derived cardiomyocytes onto a myocardial infarction model in immunosuppressed rats (1.0×107 cells/rat), followed by echocardiographic evaluation of cardiac function four weeks post-transplantation (N=3-6). Results There was less leakage from the injection site at lower CP concentration (10%) compared to higher concentration (20%), particularly in chicken or ex vivo rat heart tissue under static, non-beating conditions (chicken; 10% vs. 20% CP: 3.2±0.7% vs. 11.6±0.8%, P=0.01 / rat heart; 1.5±0.9% vs. 6.8±1.1%, P=0.0072). However, when examining fluid leakage from the cross-section of ex vivo rat hearts, we observed significantly higher leakage, regardless of CP concentration (10% CP: 45.7±2.7%, 20% CP: 59.1±8.1%), suggesting that CP diffusion within myocardial tissues had a greater influence on local retention than leakage from the injection site. Subsequently, in an experiment using beating rat myocardium and Indian ink, we observed significant suppression of diffusion in tissues treated with CP solution, with the 20% concentration showing a tendency to remain at the injection site (0% CP: 24.5±6.2 mm2, 10% CP: 6.9±7.1 mm2, 20% CP: 10.6±3.3 mm2, P=0.005). This indicates that 20% CP, rather than 10% CP, may be more suitable for local retention in tissues experiencing intra-tissue pressure, such as the beating myocardium. Finally, in vivo experiment using a myocardial infarction model of rat, there was no significant difference in left ventricular ejection fraction (LVEF) when cardiomyocytes were transplanted with 10% CP (0％ CP: 56.1±8.9%, 10%CP: 61.9±5.2%, P=0.92). However, there was a trend towards improved LVEF with the 20% CP solution (0% CP: 56.1±8.9%, 20%CP: 68.8±7.9%, P=0.13), Conclusion Twenty-percent CP may be more beneficial for treating myocardial ischemia due to enhanced retention and engraftment of transplanted cardiomyocytes.

Comprehensive Pharmacokinetic Evaluation of High Melanin Binder Levofloxacin in Rabbits Shows Potential of Topical Eye Drops for Posterior Segment Treatment.

The purpose of this work was to understand the impact of melanin binding on ocular pharmacokinetics after administration of a high-binder model drug via different administration routes. We applied levofloxacin to pigmented and albino rabbits as eye drops (single and multiple), as well as by intravitreal and intravenous injections. Ocular tissues and plasma were analyzed for levofloxacin concentrations with liquid chromatography-mass spectrometry (LC-MS/MS), and pharmacokinetic parameters were calculated. The data show enrichment of levofloxacin and weeks-long retention in pigmented tissues. Upon intravitreal injection, the area under the curve (AUC) values in pigmented tissues were about 9 to 15 times higher than the respective values in the albino rabbits, but this difference expanded to 255- to 951-fold following topical eye drop administration. Multiple dosing of eye drops led to substantial accumulation of levofloxacin in the pigmented tissues: AUC values were 3 to 12 times higher than after intravitreal injection. The AUCs were much lower after single topical or intravenous drug administrations. High drug levels (0.1-35 µM) were always observed in the neural retinas of pigmented eyes; the highest exposure was seen after intravitreal administration followed by multiple doses of topical drops. Single topical instillation and intravenous injections to the albino rabbits resulted in vitreal bioavailability values of 0.009% and 0.003%, respectively. Melanin binding can be used to achieve targeted drug delivery and extended retention in pigmented ocular tissues. The results from topical multiple dosing experiments suggest that eye drop treatment may yield drug exposures and responses comparable to intravitreal delivery, even in the retinal pigment epithelium and choroid.

8041 Eneboparatide, A Potent Stimulator Of Urinary Calcium Reabsorption, Induces Prolonged Renal Cortex Retention And PTH1 Receptor Signaling

Abstract Disclosure: G. Ravel: Employee; Self; Amolyt Pharma. R. Datta: Consulting Fee; Self; Amolyt Pharma. S. Milano: Employee; Self; Amolyt Pharma. M. Ovize: Employee; Self; Amolyt Pharma. S. Allas: Employee; Self; Amolyt Pharma. M. Aouadi: None. M.D. Culler: Employee; Self; Amolyt Pharma. Natural parathyroid hormone (PTH) replacement therapy for chronic hypoparathyroidism (cHP) is not ideal due to both short circulating half-life and brief receptor signaling, making the effect too transient. Following agonist binding to the PTH 1 receptor (PTH1R), the magnitude and duration of the receptor signal, and consequently the biological response, appears to depend on the affinity of the ligand for a specific receptor conformation termed R0, which determines the duration of ligand binding. Eneboparatide (Enebo) is a hybrid analog of PTH and PTH-related peptide designed to strongly bind to the R0 conformation while having a short circulating half-life similar to that of PTH. When tested in cHP patients, Enebo eliminated the need for calcium (Ca) (88% patients) and Vitamin D (92% patients) supplementation, while maintaining serum Ca levels over a full 24 hours. Urinary calcium (uCa) was rapidly decreased and was normalized in 92% of patients who were hypercalciuric prior to treatment. To explore the mechanism of the potent effect of Enebo on uCa, a binding assay specific for the R0 conformation of the PTH1R confirmed the high affinity of Enebo versus natural PTH1-34, with IC50 = 1.5nM and 30.9nM, respectively. Postulating that the high-affinity binding of Enebo to R0 should result in longer tissue retention in vivo, we examined the tissue levels of PTH1-34 and Enebo in male, 200-300g rats using quantitative whole-body autoradiography (QWBA) following subcutaneous injection of 3H-labeled Enebo and PTH1-34. Tissue levels of the compounds were normalized based on the concentrations observed in cardiac blood. The highest level observed for both compounds was in the renal cortex, the major site of active PTH-induced Ca reabsorption, with peak concentrations observed 30 minutes post-injection. Six hours after injection, 96.3% of PTH1-34 had disappeared from the renal cortex, as compared with only 61.4% of Enebo, despite both compounds having similar circulating half-lives. With respect to activating the PTH1R, both compounds showed a similar increase in cAMP in human embryonic kidney (HEK293) cells transfected with the PTH1R and the Green Downward cADDis cAMP biosensor, with EC50 of 0.32 and 0.48nM for Enebo and PTH1-34, respectively. To mimic the effect of rapid elimination in vivo, the ligands were added to the cells, washed-off after 15 minutes, and cAMP production followed over the next 16 hours. After washout, the cAMP signal induced by PTH1-34 rapidly returned to baseline in less than 60 minutes, whereas the cAMP signal induced by Enebo gradually decreased and was still evident after 16 hours. Collectively, these data confirm high affinity of Enebo for the R0 conformation of the PTH1R, as well as prolonged receptor signaling and retention by the renal cortex, clearly differentiating Enebo from PTH1-34 and providing strong rationale for the efficacy of Enebo in reducing uCa in cHP patients. Presentation: 6/3/2024

Janus-Structured Microgel Barrier with Tissue Adhesive and Hemostatic Characteristics for Efficient Prevention of Postoperative Adhesion.

Postoperative adhesion (POA) is a common and serious complication following various types of surgery. Current physical barriers either have a short residence time at the surgical site with a low tissue attachment capacity or are prone to undesired adhesion formation owing to the double-sided adhesive property, which limits the POA prevention efficacy of the barriers. In this study, Janus-structured microgels (Janus-MGs) with asymmetric tissue adhesion capabilities are fabricated using a novel bio-friendly gas-shearing microfluidic platform. The anti-adhesive side of Janus-MGs, which consists of alginate, hyaluronic acid, and derivatives, endows the material with separation, lubrication, and adhesion prevention properties. The adhesive side provided Janus-MGs with tissue attachment and retention capability through catechol-based adhesion, thereby enhancing the in situ adhesion prevention effect. In addition, Janus-MGs significantly reduced blood loss and shortened the hemostatic time in rats, further reducing adhesion formation. Three commonly used rat POA models with different tissue structures and motion patterns are established in this study, namely peritoneal adhesion, intrauterine adhesion, and peritendinous adhesion models, and the results showed that Janus-MGs effectively prevented the occurrence of POA in all the models. The fabrication of Janus-MGs offers a reliable strategy and a promising paradigm for preventing POA following diverse surgical procedures.

Skeletal Muscle Memory: An Update From the Antidoping Perspective.

This narrative review explores the concept of muscle memory, focusing on the physiological and biochemical mechanisms underlying information retention in skeletal muscle tissue as it relates to antidoping. The discussion encompasses the role of satellite cells (SCs) in myonuclei recruitment, resulting in increased myonuclear density and heightened muscle protein turnover. The myonuclear domain theory suggests that myonuclei acquired during hypertrophy may persist, contributing to enhanced muscle protein synthesis (MPS) and potential benefits of muscle memory. The impact of sustained training, protein intake, and resistance exercise on muscle memory, especially in elite athletes, is considered. The review also delves into the influence of anabolic androgenic steroids (AAS) on muscle tissue, highlighting their role in elevating the performance threshold and supporting recovery during intense training through increased muscle protein turnover rates. Additionally, genetic and epigenetic modifications, such as DNA methylation, are explored as potential contributors to muscle memory. The complex interplay of continuous training, AAS use, and genetic factors offers avenues for further research, especially in the context of antidoping efforts. The understanding of muscle memory has implications for maintaining performance gains and addressing ethical challenges in sports.

20 Trace metal homeostasis in growing cattle

Abstract Apparent trace metal absorption and tissue retention upon incremental levels of supplemental Zn, Cu, and Mn were studied in growing cattle. A total of 60 Holstein bulls were enrolled for the study and fed for 28 d a diet consisting of barley straw (15%), molasses (10%) and pelleted concentrate (75%; Zn = 78 ppm, Cu = 15 ppm, Mn = 91 ppm). Thereafter, 20 bulls were randomly selected and slaughtered for determination of baseline tissue trace metal composition. The remaining 40 bulls [body weight (BW) = 394 ± 20 kg] were then blocked based on BW in 8 blocks of 5 bulls each. Bulls within a block were randomly assigned one of the diets differing in supplemental levels of Zn, Cu, and Mn (all from sulfate form) with the following dietary contents of Zn, Cu and Mn, respectively: No supplement (38, 7, and 47 ppm), Dose 1 (61, 11, and 68 ppm), Dose 2 (78, 15 and 91 ppm), Dose 3 (123, 26, 136 ppm) or Dose 4 (195, 43, and 214 ppm). These diets were fed for 12 wk, and complete collection of feces and urine were performed on wk 4, 8 and 12 for assessment of trace metals balance and apparent absorption. Gastrointestinal tissues, whole organs (heart, liver, spleen, and kidney), bile, cervical vertebra, and tibia were collected at slaughterhouse from all bulls. All samples were dried, ground, and analyzed for minerals. Data were processed using dietary treatment as a continuous variable and also as categorial variable. Both approaches adjusted for the fixed effect of block (Proc Mixed, SAS 9.4). Whole-body Zn, Cu and Mn balance quadratically increased (P < 0.05) with supplementation, with a significantly higher values for Dose 4 compared with all other dietary treatment (Figure 1, P < 0.05). Dose 4 also led to a 2-fold increase in urinary Zn and Cu excretions (P < 0.05), and a 4-fold increase in urinary Mn excretion (P < 0.01). Although magnitudes of change were moderate (8 to 15%), increasing dietary Zn supply linearly increased spleen Zn content (P = 0.03), and tended to increase ruminal and hepatic Zn content (P < 0.10). Hepatic Cu linearly increased from 264 to 667 mg/kg DM with increasing dietary Cu supply (P < 0.01), whereas Cu concentration in bile only increased at Doses 4 and 5 (P < 0.01). Increasing Mn supply resulted in quadratic increase of ruminal and biliary Mn content from 340 to 1140 mg/kg DM, and from 0.32 to 1.38 mg/L, respectively. Small intestine and cecum Mn concentrations linearly and quadratically increased (about 2-fold increase, P < 0.05) with incremental supply of Mn. Homeostatic regulation mechanisms were apparently overwhelmed at the greatest Zn, Cu and Mn supplementation levels evaluated in this study.

Supramolecular Enzyme-Peptide Gels for Localized Therapeutic Biocatalysis.

Enzyme therapeutics are often compromised by poor accumulation within target tissues, necessitating high doses that can exacerbate off-target effects. We report an injectable supramolecular enzyme-peptide gel platform for prolonged local enzyme retention in vivo . The gel is based on CATCH(+/-) ( C o- A ssembling T ags based on CH arge-complementarity), cationic and anionic peptide pairs that form β-sheet fibrils upon mixing 1 . Enzymes recombinantly fused to CATCH(-) peptide integrate into CATCH(+/-) β-sheet fibrils during assembly, resulting in an enzyme-peptide gel. Catalytically-active gels were fabricated with four disparate enzymes: CATCH(-)-NanoLuc luciferase, CATCH(-)-cutinase, CATCH(-)-uricase, and CATCH(-)-adenosine synthase A. CATCH(-)-cutinase gels demonstrated tunability of the platform, while CATCH(-)-NanoLuc gels demonstrated prolonged tissue retention relative to soluble enzyme. CATCH(-)-uricase gels resolved localized inflammation in a gout model, while CATCH(-)-adenosine synthase A gels suppressed localized lipopolysaccharide-induced inflammation. Modular and tunable enzyme content, coupled with prolonged tissue retention, establish CATCH enzyme-peptide gels as a generalizable vehicle for effective local therapeutic biocatalysis.

Cell-penetrating peptides TAT and 8R functionalize P22 virus-like particles to enhance tissue distribution and retention in vivo.

Virus-like particles (VLPs) are used as nanocontainers for targeted drug, protein, and vaccine delivery. The phage P22 VLP is an ideal macromolecule delivery vehicle, as it has a large exterior surface area, which facilitates multivalent genetic and chemical modifications for cell recognition and penetration. Arginine-rich cell-penetrating peptides (CPPs) can increase cargo transport efficiency in vivo. However, studies on the tissue distribution and retention of P22 VLPs mediated by TAT and 8R are lacking. This study aimed to analyze the TAT and 8R effects on the P22 VLPs transport efficiency and tissue distribution both in vitro and in vivo. We used a prokaryotic system to prepare P22 VLP self-assembled particles and expressed TAT-or 8R-conjugated mCherry on the VLP capsid protein as model cargoes and revealed that the level of P22 VLP-mCherry penetrating the cell membrane was low. However, both TAT and 8R significantly promoted the cellular uptake efficiency of P22 VLPs in vitro, as well as enhanced the tissue accumulation and retention of P22 VLPs in vivo. At 24 h postinjection, TAT enhanced the tissue distribution and retention in the lung, whereas 8R could be better accumulation in brain. Thus, TAT was superior in terms of cellular uptake and tissue accumulation in the P22 VLPs delivery system. Understanding CPP biocompatibility and tissue retention will expand their potential applications in macromolecular cargo delivery.

Indocyanine green uptake by human tumor and non‑tumor cell lines and tissue.

Indocyanine green (ICG) is a potential promising dye for a better intraoperative tumor border definition and an improved patient outcome by potentially improving tumor border visualization compared with traditional white light guided surgery. Here, the cellular uptake of ICG in human squamous cell carcinoma (SCC026) and immortalized non-cancer skin (HaCaT) cell lines was evaluated to study the tumor-specific cellular uptake of ICG. The spatial distribution of ICG inside tumor tissue was investigated in tissue sections of head and neck squamous cell carcinoma at a microscopic level. ICG uptake and internalization was observed in living cells after 2.5 h and in the nucleus after 24 h. In dead cells, higher and faster uptake was observed. In the tissue sections, higher ICG signal intensity could be detected in connective tissue and surrounding clusters and blood vessels. In conclusion, no distinct ICG uptake by tumor cells was detected in cancer cell lines and tumor tissue. ICG localization in certain regions of tumor tissue appears to be a result of enhanced tissue permeability and retention, but not specific to tumor cells.

P12-21 GTL synthetic paraffin oil shows low liver and tissue retention compared to mineral oil – differences in MOSH interpretation

P12-21 GTL synthetic paraffin oil shows low liver and tissue retention compared to mineral oil – differences in MOSH interpretation

Comparative study of dietary selenium sources on gilthead seabream (Sparus aurata): Growth, nutrient utilization, stress response and final product quality

Selenium (Se) is a crucial trace mineral in human nutrition, known for its various health benefits, including the enhancement of antioxidant defenses. Enrichment of fish fillets with Se is seen as an interesting strategy to reduce the risk of Se deficiency in human diets. While inorganic sources have been traditionally used to supplement Se in fish diets, organic Se forms have demonstrated superior bioavailability and efficacy in managing oxidative stress and tissue retention rates compared to their inorganic counterparts.The main objective of this study was to assess the performance, stress resistance, and product quality of gilthead seabream (Sparus aurata) in response to diets supplemented with 0.2 mg kg−1 of Na2SeO3 (CTRL), Zn-L-SeMet (AVSe) or OH-Se-Met (ORGSe), complying therefore with the European Food Safety Agency (EFSA) maximum allowed supplementation of organic Se (0.2 mg kg−1). Diets were formulated to be similar in their protein (48% DM) and lipid content (17% DM), and hand-fed to apparent satiety to quadruplicate groups of 20 fish (initial body weight ∼ 75 g) gilthead seabream juveniles for 137 days. Fish were kept in a recirculating saltwater system (salinity 35‰, 22 ± 1 °C) until they reached commercial size (>250 g). At the end of the growth trial, all fish were individually weighed and measured, with 4 fish per tank sampled for whole-body composition analysis. Fillet yield, muscle lipid and mineral composition, and hepatic oxidative stress indicators were assessed (n = 12). Lipid peroxidation, muscle pH, water-holding capacity (WHC), color, and texture were evaluated on days 7 and 14 (n = 8), while sensory analysis was conducted on 20 fish. Additionally, 12 fish per treatment were subjected to overcrowding (100 kg m−3) for 5 min and air exposure for 1 min. After a 1-h recovery period, plasma and liver samples were collected from these fish, and results were compared to those of the non-stress group. All diets yielded equivalent growth performance, ensuring low feed conversion ratio. Fish fed the AVSe diet exhibited higher whole-body protein content compared to those fed the CTRL diet. Regarding mineral utilization, increased Se retention and gain in fish fed AVSe and ORGSe diets led to higher Se content in these fish compared to the CTRL. Additionally, the AVSe diet has also resulted in significantly increased Se deposition in the muscle compared to the other tested Se sources, contributing with 34% of the Se Daily Recommended Intake (DRI). The assessed flesh quality parameters and overall consumer acceptance did not reveal significant differences among the groups. Exposure to an acute stress was found to decrease the activities of Glutathione reductase and glutathione peroxidase, which in turn has increased the level of oxidized glutathione, particularly in the AVSe group. Compared to OH-Se-Met and Na2SeO3, Zn-L-SeMet was more effective in promoting response to stress and nutritional value of gilthead seabream as an important source of protein and Se in human diet.

Selenium nanoparticle inclusion in broiler diets for enhancing sustainable production and health

This study aimed to evaluate the effects of dietary supplementation of nanoparticles of Selenium (Nano-Se) on productive performance, nutrient digestibility, carcass criteria, selenium retention, blood biochemistry, and histopathological examination of broiler chicken. A total of 192 1-day-old male broiler chickens (Cobb 500) were randomly assigned to one of four treatment diets, with each diet given to six replicates of eight chicks. The birds were randomly assigned to one of four treatment groups, each of which included Nano-Se at levels of 0, 0.2, 0.3, or 0.4 mg/kg. The feeding experiment lasted 35 days. Nano-Se addition to broiler diets at 0.2 and 0.3 mg/kg enhanced body weight and body weight gain linearly compared to the control diet and 0.4 mg/kg. The apparent digestibility coefficient of ether extracts linearly increased with increasing Nano-Se levels up to 0.4 mg/kg. Increasing Nano-Se decreased serum cholesterol, triglycerides, alanine aminotransaminase, aspartate aminotransaminase, and creatinine in broiler chickens. Also, serum antioxidants showed a significant increase with increasing Nano-Se levels. As Nano-Se levels were supplemented, improvements in cooking loss, water-holding capacity, and antioxidants were observed as compared to the control. Additionally, a noticeable improvement in meat quality was observed regarding the obtained meat characters. It was preferred to use low doses of Nano-Se (0.3 mg/kg), as tissue retention of Se for both meat and liver was more comparable to the control. In conclusion, nutritional supplementation with Nano-Se increased growth performance, nutrient digestibility, selenium retention, meat quality, blood biochemistry, histological indices, and antioxidant activity of broiler chickens. Overall, the best performance of broilers was observed with Nano-Se supplementation at 0.3 mg/kg, highlighting its potential as a novel supplement for broiler diets.

Tissue surface adaptation and retention of digital obturator after one year of use

BackgroundEffect of aging on tissue adaptability and retention of digital obturator is still under investigation.Methods A maxillary Armany (class I) epoxy reference model was scanned to fabricate digital obturator fabricated from milled Co-Cr framework and 3D printed bulb. A color map of the scanned reference and digital obturator was made using Geomagic software to evaluate the accuracy of fit before and after cyclic loading using ROBOTA chewing simulator at 37,500, 75,000 and 150,000 cycles to simulate clinically 3-, 6- and 12-months chewing condition. Insertion-removal condition simulating the placement and removal of the obturator was done using repeated 360, 720 and 1440 cycles and retention was evaluated before and after the repeated cycles. Data were collected, tabulated and statistically analyzed using Statistical Package for Social Sciences (IBM SPSS Statistics 26). Student t-test and multi variable ANOVA test were used to detect significance. P-value < 0.05 was considered significant difference.ResultsFor retention test: There was a significant difference between baseline and 3, 6 and12 months. For the tissue surface adaptation test: There was significant difference at all measured areas (P-value < 0.05) before and after application of load.Conclusiondigitally designed and fabricated obturator was highly retentive and has excellent tissue surface adaptation upon fabrication, After application of load; reduction of retention and lack of tissue adaptation were resulted.The clinical implicationof this manuscript is that digital obturator can be used successfully with the shortcomings of loosening retention and adaptation afterwhile. So, clinical trials should investigate the clinical acceptance of these shortcomings.

The Arabidopsis thaliana ecotype Ct-1 achieves higher salt tolerance relative to Col-0 via higher tissue retention of K+ and NO3-

Agriculture is vital for global food security, and irrigation is essential for improving crop yields. However, irrigation can pose challenges such as mineral scarcity and salt accumulation in the soil, which negatively impact plant growth and crop productivity. While numerous studies have focused on enhancing plant tolerance to high salinity, research targeting various ecotypes of Arabidopsis thaliana has been relatively limited. In this study, we aimed to identify salt-tolerant ecotypes among the diverse wild types of Arabidopsis thaliana and elucidate their characteristics at the molecular level. As a result, we found that Catania-1 (Ct-1), one of the ecotypes of Arabidopsis, exhibits greater salt tolerance compared to Col-0. Specifically, Ct-1 exhibited less damage from reactive oxygen species (ROS) than Col-0, despite not accumulating antioxidants like anthocyanins. Additionally, Ct-1 accumulated more potassium ions (K+) in its shoots and roots than Col-0 under high salinity, which is crucial for water balance and preventing dehydration. In contrast, Ct-1 plants were observed to accumulate slightly lower levels of Na+ than Col-0 in both root and shoot tissues, regardless of salt treatment. These findings suggest that Ct-1 plants achieve high salinity resistance not by extruding more Na+ than Col-0, but rather by absorbing more K+ or releasing less K+. Ct-1 exhibited higher nitrate (NO3-) levels than Col-0 under high salinity conditions, which is associated with enhanced retention of K+ ions. Additionally, genes involved in NO3- transport and uptake, such as NRT1.5 and NPF2.3, showed higher transcript levels in Ct-1 compared to Col-0 when exposed to high salinity. However, Ct-1 did not demonstrate significantly greater resistance to osmotic stress compared to Col-0. These findings suggest that enhancing plant tolerance to salt stress could involve targeting the cellular processes responsible for regulating the transport of NO3- and K+. Overall, our study sheds light on the mechanisms of plant salinity tolerance, emphasizing the importance of K+ and NO3- transport in crop improvement and food security in regions facing salinity stress.

Abstract We060: Hypoxia Inducible Factor-2α Enhances Neutrophil Survival and Promotes Cardiac Injury Following Myocardial Infarction via cIAP-1 Signaling

Rationale: Heart failure is a major cause of mortality following myocardial infarction (MI). Neutrophils (PMNs) are among the first immune cells to accumulate in the infarcted region. While some long-term beneficial functions of PMN in heart injury are now appreciated, en mase PMN accumulation in injured hearts is still associated with worsened resolution of inflammation and disease outcomes. MI causes extensive hypoxic injury to heart tissue, where hypoxia-inducible factors (HIFs) play critical roles in cellular functions. While HIF1α regulation of cell survival, migration and metabolism have been well-studied, the role of HIF-2α in these processes is not well-understood. Even less is known of HIF-2α contributions to PMN effector functions, especially in the context of MI. Objectives: The current study aimed to 1. Mechanistically explore how HIF-2α regulates survival of tissue infiltrating PMNs post-MI and 2. Determine how HIF-2α activity in PMNs impacts heart injury resolution post-MI, and whether specific HIF-2α deletion in PMNs can alleviate MI-induced tissue injury. Methods &amp; Results: Infarct size, cardiac functions monitored by echocardiography, and scar tissue formation after left anterior descending coronary ligation injury model were compared between littermate control and specific PMN HIF-2α depletion (Ly6G-cre +/- HIF-2α fl/fl ) mice. Myocardial infarction size, cardiac dysfunction, scare tissue deposit and survival of tissue infiltrating PMNs post-MI were substantially attenuated in mice with PMN HIF-2α deletion. A significantly reduced lifespan of HIF-2α knockout PMNs was further confirmed by fate-mapping EdU pulse chase experiments. Mechanistically, molecular studies revealed that HIF-2α transcriptionally regulates expression of the pro-survival factor cIAP1 in PMNs binding to an HRE located in the promoter region of BIRC2 , in a hypoxia-inducible manner. Ex-vivo pharmacological inhibition of cIAP1 expression in PMNs using Birinapant resulted in dramatic cell death, establishing the critical role of cIAP1 in PMN survival. Finally, our data demonstrate that the protective effect of HIF-2α deletion in PMNs on MI outcomes was due to elevated recruitment of reparative macrophages into infarcted hearts. Conclusion: HIF-2α, through cIAP1 regulation, promotes PMN survival and tissue retention post-MI, leading to worsen cardiac functions and scar formation.

IL-1ra loaded chondroitin sulfate-functionalized microspheres for minimally invasive treatment of intervertebral disc degeneration

Presently, the clinical treatment of intervertebral disc degeneration (IVDD) remains challenging, but the strategy of simultaneously overcoming the overactive inflammation and restoring the anabolic/catabolic balance of the extracellular matrix (ECM) in the nucleus pulposus (NP) has become an effective way to alleviate IVDD. IL-1ra, a natural antagonist against IL-1β, can mitigate inflammation and promote regeneration in IVDD. Chondroitin sulfate (CS), an important component of the NP, can promote ECM synthesis and delay IVDD. Thus, these were chosen and integrated into functionalized microspheres to achieve their synergistic effects. First, CS-functionalized microspheres (GelMA-CS) with porous microstructure, good monodispersion, and about 200 µm diameter were efficiently and productively fabricated using microfluidic technology. After lyophilization, the microspheres with good local injection and tissue retention served as the loading platform for IL-1ra and achieved sustained release. In in vitro experiments, the IL-1ra-loaded microspheres exhibited good cytocompatibility and efficacy in inhibiting the inflammatory response of NP cells induced by lipopolysaccharide (LPS) and promoting the secretion of ECM. In in vivo experiments, the microspheres showed good histocompatibility, and local, minimally invasive injection of the IL-1ra-loaded microspheres could reduce inflammation, maintain the height of the intervertebral disc (IVD) and the water content of NP close to about 70 % in the sham group, and retain the integrated IVD structure. In summary, the GelMA-CS microspheres served as an effective loading platform for IL-1ra, eliminated inflammation through the controlled release of IL-1ra, and promoted ECM synthesis via CS to delay IVDD, thereby providing a promising intervention strategy for IVDD. Statement of significanceThe strategy of simultaneously overcoming the overactive inflammation and restoring the anabolic/catabolic balance of the extracellular matrix (ECM) in nucleus pulposus (NP) has shown great potential prospects for alleviating intervertebral disc degeneration (IVDD). From the perspective of clinical translation, this study developed chondroitin sulfate functionalized microspheres to act as the effective delivery platform of IL-1ra, a natural antagonist of interleukin-1β. The IL-1ra loading microspheres (GelMA-CS-IL-1ra) showed good biocompatibility, good injection with tissue retention, and synergistic effects of inhibiting the inflammatory response induced by lipopolysaccharide and promoting the secretion of ECM in NPCs. In vivo, they also showed the beneficial effect of reducing the inflammatory response, maintaining the height of the intervertebral disc and the water content of the NP, and preserving the integrity of the intervertebral disc structure after only one injection. All demonstrated that the GelMA-CS-IL-1ra microspheres would have great promise for the minimally invasive treatment of IVDD.

Differential response of proline metabolism defense, Na+ absorption and deposition to salt stress in salt-tolerant and salt-sensitive rapeseed (Brassica napus L.) genotypes.

Soil salinization is a major abiotic factor threatening rapeseed yields and quality worldwide, yet the adaptive mechanisms underlying salt resistance in rapeseed are not clear. Therefore, this study aimed to explore the differences in growth potential, sodium (Na+) retention in different plant tissues, and transport patterns between salt-tolerant (HY9) and salt-sensitive (XY15) rapeseed genotypes, which cultivated in Hoagland's nutrient solution in either the with or without of 150 mM NaCl stress. The results showed that the inhibition of growth-related parameters of the XY15 genotype was higher than those of the HY9 in response to salt stress. The XY15 had lower photosynthesis, chloroplast disintegration, and pigment content but higher oxidative damage than the HY9. Under NaCl treatment, the proline content in the root of HY9 variety increased by 8.47-fold, surpassing XY15 (5.41-fold). Under salt stress, the HY9 maintained lower Na+ content, while higher K+ content and exhibited a relatively abundant K+/Na+ ratio in root and leaf. HY9 also had lower Na+ absorption, Na+ concentration in xylem sap, and Na+ transfer factor than XY15. Moreover, more Na+ contents were accumulated in the root cell wall of HY9 with higher pectin content and pectin methylesterase (PME) activity than XY15. Collectively, our results showed that salt-tolerant varieties absorbed lower Na+ and retained more Na+ in the root cell wall (carboxyl group in pectin) to avoid leaf salt toxicity and induced higher proline accumulation as a defense and antioxidant system, resulting in higher resistance to salt stress, which provides the theoretical basis for screening salt resistant cultivars.