Complement networks in gene-edited pig xenotransplantation: enhancing transplant success and addressing organ shortage.

The shortage of organs for transplantation emphasizes the urgent need for alternative solutions. Xenotransplantation has emerged as a promising option due to the greater availability of donor organs. However, significant hurdles such as hyperacute rejection and organ ischemia-reperfusion injury pose major challenges, largely orchestrated by the complement system, and activated immune responses. The complement system, a pivotal component of innate immunity, acts as a natural barrier for xenotransplantation. To address the challenges of immune rejection, gene-edited pigs have become a focal point, aiming to shield donor organs from human immune responses and enhance the overall success of xenotransplantation. This comprehensive review aims to illuminate strategies for regulating complement networks to optimize the efficacy of gene-edited pig xenotransplantation. We begin by exploring the impact of the complement system on the effectiveness of xenotransplantation. Subsequently, we delve into the evaluation of key complement regulators specific to gene-edited pigs. To further understand the status of xenotransplantation, we discuss preclinical studies that utilize gene-edited pigs as a viable source of organs. These investigations provide valuable insights into the feasibility and potential success of xenotransplantation, offering a bridge between scientific advancements and clinical application.

Advantages of the retroperitoneal retrocolic space as the transplant site for encapsulated xenogeneic islets.

OBJECTIVE: Islet allotransplantation has demonstrated improved clinical outcomes using the hepatic portal vein as the standard infusion method. However, the current implantation site is not ideal due to the short-term thrombotic and long-term immune destruction. Meanwhile, the shortage of human organ donors further limits its application. To find a new strategy, we tested a new polymer combination for islet encapsulation and transplantation. Meanwhile, we explored a new site for xenogeneic islet transplantation in mice. METHOD: We synthesized a hydrogel combining alginate plus poly-ethylene-imine (Alg/PEI) for the encapsulation of rat, neonatal porcine, and human islets. Transplantation was performed into the retroperitoneal retro-colic space of diabetic mice. Control mice received free islets under the kidney capsule or encapsulated islets into the peritoneum. The biochemical indexes were measured, and the transplanted islets were harvested for immunohistochemical staining of insulin and glucagon. RESULTS: Mice receiving encapsulated rat, porcine and human islets transplanted into the retroperitoneal space maintained normoglycemia for a median of 275, 145.5, and 146 days, respectively. In contrast, encapsulated xenogeneic islets transplanted into the peritoneum, maintained function for a median of 61, 95.5, and 82 days, respectively. Meanwhile, xenogeneic islets transplanted free into the kidney capsule lost their function within 3 days after transplantation. Immunohistochemical staining of encapsulated rat, porcine and human islets, retrieved from the retroperitoneal space, allowed to distinguish morphological normal insulin expressing ß- and glucagon expressing α-cells at 70, 60, and 100 days post-transplant, respectively. CONCLUSION: Transplantation of Alg/PEI encapsulated xenogeneic islets into the retroperitoneal space provides a valuable new implantation strategy for the treatment of type 1 diabetes.

Observing the Viscous Relaxation Process of Silica Optical Fiber at ~1000 °C Using Regenerated Fiber Bragg Grating.

A regenerated fiber Bragg grating (RFBG) in silica fiber was used to observe the viscous relaxation process of the host silica fiber at high temperatures of around 1000 °C. Two factors, preannealing time and loaded tension, which affect viscous relaxation, were observed. When an RFBG is stretched after a longer preannealing, the measured viscosity of the optical fiber was observed to reach equilibrium faster, which means that preannealing accelerates viscous relaxation. A similar acceleration phenomenon was also observed when a larger load was applied to stretch the optical fiber, although the acceleration effect of loaded tension was not as strong as in the preannealing case. The results play an active role in establishing effective optical-fiber devices for application in high-temperature environments.

Electric-arc-induced strength-controllable weak polarization mode coupling in polarization maintaining fiber.

An electric-arc-based scheme to generate strength-controllable weak polarization mode coupling (PMC) points in polarization maintaining fiber (PMF) is described. The resulting PMC strengths can be readily controlled to be in the very weak range of -60 to -40 dB. In this range, excellent mechanical strength combined with high return loss is achieved. An experimental quasi-distributed temperature sensor is formed by three separate PMC points in a single PMF using the electric arc method.

Polarization mode coupling and related effects in fiber Bragg grating inscribed in polarization maintaining fiber.

Polarization mode coupling (PMC) and related effects from writing fiber Bragg gratings in polarization maintaining fiber (FBGs-in-PMF) are observed experimentally for the first time by optical fiber coherence domain polarimetry (OCDP) using a broadband light source. PMC is another useful aspect of FBG-in-PMF besides Bragg wavelength and its possible potential is evaluated and discussed. A localized and long range temperature measurement based on the PMC and Bragg wavelength is given as an example.

KIF2A Upregulates PI3K/AKT Signaling through Polo-like Kinase 1 (PLK1) to Affect the Proliferation and Apoptosis Levels of Eriocheir sinensis Spermatogenic Cells.

E. sinensis is an animal model for studying the reproduction and development of crustaceans. In this study, we knocked down the Es-Kif2a gene by injecting dsRNA into E. sinensis and inhibited Es-Plk1 gene expression by injecting PLK1 inhibitor BI6727 into E. sinensis. Then, the cell proliferation level, apoptosis level, and PI3K/AKT signaling expression level were detected. Our results showed that the proliferation level of spermatogenic cells decreased, while the apoptosis level increased after Es-Kif2a knockdown or Es-Plk1 inhibition. In order to verify whether these changes are caused by regulating the PI3K/AKT pathway, we detected the expression of PI3K and AKT proteins after Es-Kif2a knockdown or Es-Plk1 inhibition. Western Blot showed that in both the Es-Kif2a knockdown group and the Es-Plk1 inhibition group, the expression of PI3K and AKT proteins decreased. In addition, immunofluorescence showed that Es-KIF2A and Es-PLK1 proteins were co-localized during E. sinensis spermatogenesis. To further explore the upstream and downstream relationship between Es-KIF2A and Es-PLK1, we detected the expression level of Es-PLK1 after Es-Kif2a knockdown as well as the expression level of Es-KIF2A after Es-Plk1 inhibition. Western Blot showed that the expression of Es-PLK1 decreased after Es-Kif2a knockdown, while there was no significant change of Es-KIF2A after Es-Plk1 inhibition, indicating that Es-PLK1 may be a downstream factor of Es-KIF2A. Taken together, these results suggest that Es-KIF2A upregulates the PI3K/AKT signaling pathway through Es-PLK1 during the spermatogenesis of E. sinensis, thereby affecting the proliferation and apoptosis levels of spermatogenic cells.

Es-ß-CATENIN affects the hemolymph-testes barrier in Eriocheir sinensis by disrupting cell junctions and cytoskeleton.

ß-CATENIN is an evolutionarily conserved multifunctional molecule that maintains cell adhesion as a cell junction protein to safeguard the integrity of the mammalian blood-testes barrier, and also regulates cell proliferation and apoptosis as a key signaling molecule in the WNT/ß-CATENIN signaling pathway. In the crustacean Eriocheir sinensis, Es-ß-CATENIN has been shown to be involved in spermatogenesis, but the testes of E. sinensis have large and well-defined structural differences from those of mammals, and the impact of Es-ß-CATENIN in them is still unknown. In the present study, we found that Es-ß-CATENIN, Es-α-CATENIN and Es-ZO-1 interact differently in the testes of the crab compared to mammals. In addition, defective Es-ß-CATENIN resulted in increased Es-α-CATENIN protein expression levels, distorted and deformed F-ACTIN, and disturbed localization of Es-α-CATENIN and Es-ZO-1, leading to loss of hemolymph-testes barrier integrity and impaired sperm release. In addition to this, we also performed the first molecular cloning and bioinformatics analysis of Es-AXIN in the WNT/ß-CATENIN pathway to exclude the effect of the WNT/ß-CATENIN pathway on the cytoskeleton. In conclusion, Es-ß-CATENIN participates in maintaining the hemolymph-testes barrier in the spermatogenesis of E. sinensis.

SEMA4D/VEGF surface enhances endothelialization by diminished-glycolysis-mediated M2-like macrophage polarization.

Cardiovascular disease remains the leading cause of death and morbidity worldwide. Inflammatory responses after percutaneous coronary intervention led to neoathrosclerosis and in-stent restenosis and thus increase the risk of adverse clinical outcomes. In this work, a metabolism reshaped surface is engineered, which combines the decreased glycolysis promoting, M2-like macrophage polarization, and rapid endothelialization property. Anionic heparin plays as a linker and mediates cationic SEMA4D and VEGF to graft electronically onto PLL surfaces. The system composed by anticoagulant heparin, immunoregulatory SEMA4D and angiogenic VEGF endows the scaffold with significant inhibition of platelets, fibrinogen and anti-thrombogenic properties, also noteworthy immunometabolism reprogram, anti-inflammation M2-like polarization and finally leading to rapid endothelializaiton performances. Our research indicates that the immunometabolism method can accurately reflect the immune state of modified surfaces. It is envisioned immunometabolism study will open an avenue to the surface engineering of vascular implants for better clinical outcomes.

High and Economical Nattokinase Production with Acetoin as a Useful Byproduct from Soybean Milk and Glucose.

Nattokinase (NK) is a potent fibrinolytic enzyme with wide pharmaceutical and nutraceutical applications. Safe and high NK-yielding strains are urgently needed. In this study, the best strain NDF was isolated from one of the 11 natto samples and then identified as Bacillus subtilis. The effects of carbon and nitrogen sources on NK production were investigated, and glucose and soybean milk were finally selected as the optimal carbon and nitrogen sources, respectively. Acetoin, a valuable compound with versatile usages, was detected as the main byproduct of carbon overflow. In a 6-L fermenter, NK and acetoin reached their peak concentrations simultaneously (10,220 IU/mL and 25.9 g/L, respectively) at 25 h in a culture medium containing 180 g/L of soybean milk and 105 g/L of glucose. The NK product was verified by sequencing of the aprN gene and SDS-PAGE analysis. Only very limited kinds of proteins were found in the supernatant of the fermentation broth, and NK was one of the main bands. This study has developed an economical and high NK production method with acetoin as a useful byproduct.

Dopamine D1 receptor agonist treatment alleviates morphine-exposure-induced learning and memory impairments.

We investigated the mechanisms by which the dopamine D1 receptor alleviates morphine-exposure-induced cognitive impairments. Rats were intraperitoneally injected with morphine in a dose-escalating manner over 10 days, and 15 min before the morphine injection on days 1, 3, 5, 7, and 9, the rats were administered the D1 receptor agonist SKF81297 or the D1 receptor antagonist SCH38933 into the midbrain periaqueductal gray (PAG). The Morris water maze was used to examine learning- and memory-related behavioral changes. Midbrain PAG toll-like receptor 4 (TLR4) and protein kinase A (PKA) protein expression was examined using western blot analysis, and cellular expression and localization of TLR4 and PKA were investigated using immunohistochemistry. Chronic morphine exposure impaired spatial learning and memory ability, and resulted in longer latency to the platform, decreased number of platform crossings, and shortened time in the effective area and the target quadrant. Chronic morphine exposure also increased TLR4 and PKA expression in the PAG. However, D1 receptor agonist treatment improved learning and memory ability; in morphine-treated rats, administration of the D1 receptor agonist SKF81297 could shorten the latency to the platform, increase the number of platform crossings, and increase the time spent in the effective area and the target quadrant. In addition, TLR4 expression decreased and PKA expression significantly increased in the PAG in these animals. In summary, administration of the dopamine D1 receptor agonist SKF81297 into the PAG alleviated morphine-exposure-induced cognitive impairments.

A dopamine D1 receptor agonist improved learning and memory in morphine-treated rats.

OBJECTIVE: The objective of this article is to study the role of the dopamine (DA) D1 receptor in the midbrain periaqueductal grey (PAG) on learning and memory in morphine-addicted rats. METHODS: DA D1 receptor agonist SKF81297 and D1 receptor antagonist SCH SCH23390 were administrated into the PAG, respectively, and the learning and memory behavioral changes of morphine addicted rats were detected by water maze. Western blot and immunohistochemistry were used to detect glutamate decarboxylase 67 (GAD67) and tyrosine receptor kinase B (TrkB) in PAG. RESULTS: D1 receptor agonist shortened the latency to platform and increased the number of platform crossings, indicating improved learning and memory ability of morphine addict rat. D1 receptor agonist increased GAD67 expression and decreased TrkB in PAG. CONCLUSION: (1) The PAG is involved in the learning and memory changes of the addicted rats; (2) the activation of DA D1 receptor will increase the GAD67, reduce the damage to peripheral neurons, and improve the learning and memory of the addicted rats; and (3) D1 receptor agonists further reduced TrkB expression in morphine-addicted rats, whereas TrkB levels deviated from changes in rat behavior.

Strong Antibacterial Polydopamine Coatings Prepared by a Shaking-assisted Method.

Strong antibacterial polydopamine (PDA) coatings prepared by a facile shaking-assisted method is reported for the first time. It was found that a minor modification made to the conventional synthesis procedure of PDA coatings, viz. replacing the static solution condition with a shaking solution condition by using a mechanical shaker, can produce the roughened polydopamine (rPDA) coatings at different substrates, e.g., glass, stainless steel, plastic, and gauze. The resulting rPDA coatings were characterized with Raman spectrum, zeta-potential analysis and contact angle measurement. The antibacterial activity of the rPDA coatings was evaluated by a shake flask test with gram-positive Staphylococcus aureus, and gram-negative Escherichia coli and Pseudomonas aeruginosa as bacteria models. Testing results revealed that, in the absence of any other antibacterial agents, the rPDA coatings exhibited remarkably enhanced antibacterial activities. In addition, such enhanced antibacterial activities of the rPDA coatings were found to be unimpaired by steam sterilization treatments.

Topical grape seed proanthocyandin extract reduces sunburn cells and mutant p53 positive epidermal cell formation, and prevents depletion of Langerhans cells in an acute sunburn model.

OBJECTIVE: The purpose of this study was to investigate whether grape seed proanthocyanidin extract (GSPE) can provide photoprotection against ultraviolet (UV) irradiation. BACKGROUND DATA: Study has shown that GSPE is a natural oxidant, and is used in many fields such as ischemia-reperfusion injury, chronic pancreatitis, and even cancer. However, the effect of GSPE on UV irradiation is as yet unknown. METHODS: Cutaneous areas on the backs of normal volunteers were untreated or treated with GSPE solutions or vehicles 30 min before exposure to two minimal erythema doses (MED) of solar simulated radiation. Cutaneous areas at different sites were examined histologically for the number of sunburn cells, or immunohistochemically for Langerhans cells and mutant p53 epidermal cells. RESULTS: On histological and immunohistochemical examination, skin treated with GSPE before UV radiation showed fewer sunburn cells and mutant p53-positive epidermal cells and more Langerhans cells compared with skin treated with 2-MED UV radiation only (p<0.001, p<0.001, and p<0.01, respectively). CONCLUSIONS: GSPE may be a possible preventive agent for photoprotection.