Ultra-Fast Vitrification: Minimizing the Toxicity of Cryoprotective Agents and Osmotic Stress in Mouse Oocyte Cryopreservation.

Globally, women have been adopting oocyte cryopreservation (OC) for fertility preservation for various reasons, such as inevitable gonadotoxic treatment for specific pathologic states and social preferences. While conventional vitrification (C-VIT) has improved the success rate of OC, challenges of possible toxicities of high-concentration cryoprotective agents and osmotic stress persist. To overcome these challenges, we evaluated the ultra-fast vitrification (UF-VIT) method, which reduces the equilibration solution stage exposure time compared to C-VIT by observing mouse oocyte intracellular organelles and embryonic development. Consequently, compared to fresh mouse oocytes, UF-VIT presented significant differences only in endoplasmic reticulum (ER) intensity and mitochondrial (MT) distribution. Meanwhile, C-VIT showed substantial differences in the survival rate, key ER and MT parameters, and embryonic development rate. UF-VIT exhibited considerably fewer negative effects on key MT parameters and resulted in a notably higher blastocyst formation rate than C-VIT. Meiotic spindle (spindle and chromosomes) morphology showed no significant changes between the groups during vitrification/warming (VW), suggesting that VW did not negatively affect the meiotic spindle of the oocytes. In conclusion, UF-VIT seems more effective in OC owing to efficient cytoplasmic water molecule extraction, osmotic stress reduction, and minimization of cell contraction and expansion amplitude, thus compensating for the drawbacks of C-VIT.

Effectiveness of community-based interventions for older adults living alone: a systematic review and meta-analysis.

OBJECTIVES: This study examined the effectiveness of community-based interventions designed for older adults living alone through a systematic review and meta-analysis. METHODS: The study incorporated 4 randomized controlled trials (RCTs) and 5 non-RCTs to evaluate various interventions. The methodological quality of these studies was assessed using the Downs and Black checklist, while Q-statistic and I-square tests were performed to examine statistical heterogeneity. Additionally, visual inspection of funnel plots and the trim-and-fill method were employed to investigate potential publication bias. Of the 2,729 identified studies, 9 met the criteria for inclusion in this review. Independent variables were categorized into 5 groups (physical activity, nutrition, social relationships, social participation, and combined intervention) to examine their effects. Dependent variables were similarly classified into 5 subgroups to identify the specific effects of the interventions. RESULTS: Interventions focusing on nutrition and combined approaches were the most effective, yielding effect sizes of 0.96 (95% confidence interval [CI], 0.66 to 1.25) and 0.43 (95% CI, 0.26 to 0.60), respectively. The interventions had the greatest impacts on the health behavior and mental health of the participants, with effect sizes of 0.98 (95% CI, 0.73 to 1.22) for health behavior and 0.67 (95% CI, 0.19 to 1.16) for mental health. CONCLUSIONS: This study suggests a direction for the development of community-based interventions tailored to the needs of older adults living alone. Additionally, it provides evidence to inform policy decisions concerning this demographic.

Recognition of atomic-level difference in porphyrin dyads for self-sorted supramolecular polymer growth.

Porphyrin dyads (PDMs, where M = Zn and Cu) composed of diphenylporphyrin and tetraphenylporphyrin units, designated as DPDMs and TPDMs, respectively, exhibited remarkable differences in the molecular assemblies depending on the coordination metal ion. Furthermore, TPDMs showed self-sorting behavior during the formation of supramolecular assemblies through the recognition of atomic-level difference.

Development of an In Vitro Model for Inflammation Mediated Renal Toxicity Using 3D Renal Tubules and Co-Cultured Human Immune Cells.

BACKGROUND: The emergence of various infectious diseases and the toxic effects of hyperinflammation by biotherapeutics have highlighted the need for in vitro preclinical models mimicking the human immune system. In vitro models studying the relationship between hyperinflammation and acute renal injury mainly rely on 2D culture systems, which have shown limitations in recapitulating kidney function. Herein, we developed an in vitro kidney toxicity model by co-culturing 3D engineered kidney proximal tubules cells (RPTEC/TERT1) with human peripheral blood mononuclear cells (PBMC). METHODS: RPTEC/TERT1 were sandwich cultured to form 3D renal tubules for 16 days. The tubules were then co-cultured with PBMC using transwell (0.4 µm pores) for 24 h. Hyperinflammation of PBMC was induced during co-culture using polyinosinic-polycytidylic acid (polyI:C) and lipopolysaccharide (LPS) to investigate the effects of the induced hyperinflammation on the renal tubules. RESULTS: Encapsulated RPTEC/TERT1 cells in Matrigel exhibited elevated renal function markers compared to 2D culture. The coexistence of PBMC and polyI:C induced a strong inflammatory response in the kidney cells. This hyperinflammation significantly reduced primary cilia formation and upregulated kidney injury markers along the 3D tubules. Similarly, treating co-cultured PBMC with LPS to induce hyperinflammation resulted in comparable inflammatory responses and potential kidney injury. CONCLUSION: The model demonstrated similar changes in kidney injury markers following polyI:C and LPS treatment, indicating its suitability for detecting immune-associated kidney damage resulting from infections and biopharmaceutical applications.

Real-Time, In Situ Imaging of Macrophages via Phase-Change Peptide Nanoemulsions.

Macrophages are specialized phagocytes that play central roles in immunity and tissue repair. Their diverse functionalities have led to an evolution of new allogenic and autologous macrophage products. However, realizing the full therapeutic potential of these cell-based therapies requires development of imaging technologies that can track immune cell migration within tissues in real-time. Such innovations will not only inform treatment regimens and empower interpretation of therapeutic outcomes but also enable prediction and early intervention during adverse events. Here, phase-changing nanoemulsion contrast agents are reported that permit real-time, continuous, and high-fidelity ultrasound imaging of macrophages in situ. Using a de novo designed peptide emulsifier, liquid perfluorocarbon nanoemulsions are prepared and show that rational control over interfacial peptide assembly affords formulations with tunable acoustic sensitivity, macrophage internalization, and in cellulo stability. Imaging experiments demonstrate that emulsion-loaded macrophages can be readily visualized using standard diagnostic B-mode and Doppler ultrasound modalities. This allows on-demand and long-term tracking of macrophages within porcine coronary arteries, as an exemplary model. The results demonstrate that this platform is poised to open new opportunities for non-invasive, contrast-enhanced imaging of cell-based immunotherapies in tissues, while leveraging the low-cost, portable, and safe nature of diagnostic ultrasound.

NMR-guided directed evolution.

Directed evolution is a powerful tool for improving existing properties and imparting completely new functionalities to proteins1-4. Nonetheless, its potential in even small proteins is inherently limited by the astronomical number of possible amino acid sequences. Sampling the complete sequence space of a 100-residue protein would require testing of 20100 combinations, which is beyond any existing experimental approach. In practice, selective modification of relatively few residues is sufficient for efficient improvement, functional enhancement and repurposing of existing proteins5. Moreover, computational methods have been developed to predict the locations and, in certain cases, identities of potentially productive mutations6-9. Importantly, all current approaches for prediction of hot spots and productive mutations rely heavily on structural information and/or bioinformatics, which is not always available for proteins of interest. Moreover, they offer a limited ability to identify beneficial mutations far from the active site, even though such changes may markedly improve the catalytic properties of an enzyme10. Machine learning methods have recently showed promise in predicting productive mutations11, but they frequently require large, high-quality training datasets, which are difficult to obtain in directed evolution experiments. Here we show that mutagenic hot spots in enzymes can be identified using NMR spectroscopy. In a proof-of-concept study, we converted myoglobin, a non-enzymatic oxygen storage protein, into a highly efficient Kemp eliminase using only three mutations. The observed levels of catalytic efficiency exceed those of proteins designed using current approaches and are similar with those of natural enzymes for the reactions that they are evolved to catalyse. Given the simplicity of this experimental approach, which requires no a priori structural or bioinformatic knowledge, we expect it to be widely applicable and to enable the full potential of directed enzyme evolution.

Sex and Diet-Related Disparities in Low Handgrip Strength among Young and Middle-Aged Koreans: Findings Based on the Korea National Health and Nutrition Examination Survey (KNHANES) from 2014 to 2017.

"Possible sarcopenia" may be defined as a low muscle strength assessed by handgrip strength (HGS) by sex. We examined the sex-specific association between low handgrip strength (LGS) and dietary factors for the prevention of sarcopenia in young and middle-aged Koreans. We used data from the 2014-2017 Korea National Health and Nutrition Examination Survey of 11,635 Korean adults with LGS and normal handgrip strength (NGS). The relationship between dietary factors, e.g., nutrients, foods, and dietary patterns, and HGS was evaluated by multivariate logistic regression analyses. In men, the LGS group had a higher proportion of energy from carbohydrates and a lower proportion of energy from proteins than the NGS group. The LGS group had lower protein, niacin, phosphorus, and iron densities in their diet than the NGS group. The odds of having LGS increased as intake of vitamin B1 (odds ratio (OR) 2.916, 95% confidence interval (CI) 1.265-6.719), niacin (OR 2.286, 95% CI 1.095-4.774), phosphorus (OR 2.731, 95% CI 1.036-7.199), and iron (OR 2.591, 95% CI 1.102-6.088) decreased. In women with LGS, the odds of insufficient protein intake (OR 1.976, 95% CI 1.248-3.127) was significantly higher. This study suggests that adequate intake of protein, vitamin B1, niacin, phosphorus, and iron is beneficial for maintaining HGS.

Embedding Financial Services in Frequented, Trusted Settings: Building on Families' Pre-existing Economic Mobility Efforts.

OBJECTIVE: The aim of this study was to understand parents'/guardians' experiences and aspirations around economic mobility and their impact on the acceptability and use of financial services embedded in frequented, trusted settings such as schools and pediatric clinics. METHOD: We recruited 18 English-speaking guardians with at least 1 child enrolled in a school for low-income families and eligible for Internal Revenue Service-sponsored free tax preparation. Each participant completed a semistructured interview before tax filing; a subset completed follow-up interviews. We used grounded theory analysis. RESULTS: We developed the following theoretical model based on our key thematic findings to describe the acceptability and use of financial services within the context of guardians' lived experiences and pre-existing efforts to build economic mobility: Families experience multilevel barriers to economic mobility. Despite these barriers, guardians are proactively working to build economic mobility by empowering the next generation with knowledge, skills, and assets and resiliently pursuing economic goals. As a result, guardians will accept empowering, nonjudgmental, expert, and trustworthy financial services that contribute to their existing efforts. To move from acceptance to use, financial services must be effectively publicized, accessible, and supportive. CONCLUSION: Financial services may be more acceptable and used if they are embedded in trusted organizations and have expert, supportive staff with lived experience who empower guardians to work toward their economic goals, accessible platforms, and effective publicity. These characteristics may facilitate uptake and economic mobility. Trusted organizations serving young families can partner with financial services to test these findings and help families build economic mobility.

A Deep Learning Algorithm for Classifying Diabetic Retinopathy Using Optical Coherence Tomography Angiography.

Purpose: To develop an automated diabetic retinopathy (DR) staging system using optical coherence tomography angiography (OCTA) images with a convolutional neural network (CNN) and to verify the feasibility of the system. Methods: In this retrospective cross-sectional study, a total of 918 data sets of 3 × 3 mm2 OCTA images and 917 data sets of 6 × 6 mm2 OCTA images were obtained from 1118 eyes. A deep CNN and four traditional machine learning models were trained with annotations made by a retinal specialist based on ultra-widefield fluorescein angiography. Separately, the same images of the test data sets were independently graded by two human experts. The results of the CNN algorithm were compared with those of traditional machine learning-based classifiers and human experts. Results: The proposed CNN achieved an accuracy of 0.728, a sensitivity of 0.675, a specificity of 0.944, an F1 score of 0.683, and a quadratic weighted κ of 0.908 for a six-level staging task, which were far superior to the results of traditional machine learning methods or human experts. The CNN algorithm showed a better performance using 6 × 6 mm2 rather than 3 × 3 mm2 sized OCTA images and using combined data rather than a separate OCTA layer alone. Conclusions: CNN-based classification using OCTA images can provide reliable assistance to clinicians for DR classification. Translational Relevance: This CNN algorithm can guide the clinical decision for invasive angiography or referrals to ophthalmology specialists, helping to create more efficient diagnostic workflow in primary care settings.

Morphological features and prognostic significance of multilayered pigment epithelium detachment in age-related macular degeneration.

AIMS: To investigate the structure of multilayered pigment epithelial detachment (m-PED) in neovascular age-related macular degeneration, and its association with visual prognosis and the progression of fibrotic scars at 12 months. METHODS: We retrospectively analysed 68 eyes of 63 patients with m-PED that included a prechoroidal cleft. The compartments within m-PED were divided into neovascular tissue (layer 1), a hyper-reflective band (layer 2), and a prechoroidal cleft (layer 3). Clinical variables were compared between patients manifesting layer 2 and those who did not. Multiple regression analyses were used to find the factors related to visual outcome and fibrotic scar formation. RESULTS: Layer 2 was detected in 38 (55.9 %) of 68 eyes. With continuous treatment, the group with layer 2 showed gradual visual deterioration (p<0.001 at month 12), while the group without layer 2 showed visual improvement (p<0.001 at month 12). In the group with layer 2, the thickness of layer 2 significantly increased, and in the group without layer 2, if it formed, it increased gradually (p=0.004 at month 12). In both groups, other layers significantly decreased by month 12. The presence of layer 2 at baseline was significantly associated with a poor visual outcome (p=0.009) and fibrotic scar formation (p=0.023). CONCLUSIONS: The m-PED with layer 2 had a higher risk of fibrotic scar formation and was associated with a poor visual prognosis. Layer 2 may be an early stage precursor of a fibrotic scar.

Dichloromethane fractions of Calystegia soldanella induce S­phase arrest and apoptosis in HT­29 human colorectal cancer cells.

Calystegia soldanella is a halophyte and a perennial herb that grows on coastal sand dunes worldwide. Extracts from this plant have been previously revealed to have a variety of bioactive properties in humans. However, their effects on colorectal cancer cells remain poorly understood. In the present study, the potential biological activity of C. soldanella extracts in the colorectal cancer cell line HT­29 was examined. First, five solvent fractions [n­hexane, dichloromethane (DCM), ethyl acetate, n­butanol and water] were obtained from the crude extracts of C. soldanella through an organic solvent extraction method. In particular, the DCM fraction was demonstrated to exert marked dose­ and time­dependent inhibitory effects according to results from the cell viability assay. Data obtained from the apoptosis assay suggested that the inhibition of HT­29 cell viability induced by DCM treatment was attributed to increased apoptosis. The apoptotic rate was markedly increased in a dose­dependent manner, which was associated with the protein expression levels of apoptosis­related proteins, including increased Fas, Bad and Bax, and decreased pro­caspase­8, Bcl­2, Bcl­xL, pro­caspase­9, pro­caspase­7 and pro­caspase­3. A mitochondrial membrane potential assay demonstrated that more cells became depolarized and the extent of cytochrome c release was markedly increased in a dose­dependent manner in HT­29 cells treated with DCM. In addition, cell cycle analysis confirmed S­phase arrest following DCM fraction treatment, which was associated with decreased protein expression levels of cell cycle­related proteins, such as cyclin A, CDK2, cell division cycle 25 A and cyclin dependent kinase inhibitor 1. Based on these results, the present study suggested that the DCM fraction of the C. soldanella extract can inhibit HT­29 cell viability whilst inducing apoptosis through mitochondrial membrane potential regulation and S­phase arrest. These results also suggested that the DCM fraction has potential anticancer activity in HT­29 colorectal cells. Further research on the composition of the DCM fraction is warranted.

A Comparative Study of the Hepatoprotective Effect of Centella asiatica Extract (CA-HE50) on Lipopolysaccharide/d-galactosamine-Induced Acute Liver Injury in C57BL/6 Mice.

Acute liver failure (ALF) refers to the sudden loss of liver function and is accompanied by several complications. In a previous study, we revealed the protective effect of Centella asiatica 50% ethanol extract (CA-HE50) on acetaminophen-induced liver injury. In the present study, we investigate the hepatoprotective effect of CA-HE50 in a lipopolysaccharide/galactosamine (LPS-D-Gal)-induced ALF animal model and compare it to existing therapeutic silymarin, Lentinus edodes mycelia (LEM) extracts, ursodeoxycholic acid (UDCA) and dimethyl diphenyl bicarboxylate (DDB). Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were decreased in the CA-HE50, silymarin, LEM, UDCA and DDB groups compared to the vehicle control group. In particular, AST and ALT levels of the 200 mg/kg CA-HE50 group were significantly decreased compared to positive control groups. Lactate dehydrogenase (LDH) levels were significantly decreased in the CA-HE50, silymarin, LEM, UDCA and DDB groups compared to the vehicle control group and LDH levels of the 200 mg/kg CA-HE50 group were similar to those of the positive control groups. Superoxide dismutase (SOD) activity was significantly increased in the 100 mg/kg CA-HE50, LEM and UDCA groups compared to the vehicle control group and, in particular, the 100 mg/kg CA-HE50 group increased significantly compared to positive control groups. In addition, the histopathological lesion score was significantly decreased in the CA-HE50 and positive control groups compared with the vehicle control group and the histopathological lesion score of the 200 mg/kg CA-HE50 group was similar to that of the positive control groups. These results show that CA-HE50 has antioxidant and hepatoprotective effects at a level similar to that of silymarin, LEM, UDCA and DDB, which are known to have hepatoprotective effects; further, CA-HE50 has potential as a prophylactic and therapeutic agent in ALF.

Calystegia soldanella Extract Exerts Anti-Oxidative and Anti-Inflammatory Effects via the Regulation of the NF-κB/Nrf-2 Pathways in Mouse Macrophages.

Plant polyphenols are widely used to treat various inflammatory diseases, owing to their ability to suppress reactive oxygen species production and the expression of inflammatory cytokines. Herein, we investigated phenolic compounds from Calystegia soldanella using UPLC Q-TOF MS/MS and their antioxidative and anti-inflammatory activities were analyzed. The C. soldanella ethyl acetate fraction (CsEF) had the strongest antioxidative activity, given its high polyphenol compound content. It also exhibited anti-inflammatory effects, inhibiting the production of inflammatory cytokines such as NO, PGE2, IL-1ß, IL-6, and TNF-α in LPS-stimulated mouse macrophages. CsEF activated the nuclear transcription factor Nrf-2, thereby upregulating antioxidant enzymes such as HO-1 and NQO-1 and inhibiting NF-κB expression, which in turn, suppressed the expression of COX-2, iNOS, and inflammatory cytokines, ultimately exerting anti-inflammatory effects. Further, UPLC-Q-TOF-MS/MS was used to analyze the polyphenol compound contents in CsEF. The quercetin glycosides isoquercitrin and quercitrin were the primary flavonoid compounds, while the caffeic acid derivatives, chlorogenic acid and dicaffeoylquinic acid, were the primary phenolic acids. Thus, C. soldanella, which had only a limited use thus far as a medicinal plant, may serve as a natural medicinal resource for treating inflammatory diseases.

Anti-obesity effects of galla rhois via genetic regulation of adipogenesis.

In the present study, we investigated the effects of Galla Rhois (GR) on obesity and gene expression. We prepared a GR extract and various solvent fractions and evaluated the degree to which they inhibited adipocyte differentiation and adipogenesis in vitro. Among them, the GR ethyl acetate fraction (GE) had the lowest EC50 for adipocyte differentiation and adipogenesis and thus was selected for in vivo experiments. We induced obesity in C57BL/6 mice by providing them a high-fat diet (HFD). Then, GE (10-40 mg/kg) or orlistat (positive control, 4 mg/kg) was orally administered daily for six weeks. Mean body weights and weight gain were significantly lower in the GE40 group (40 mg/kg of GE) compared with the HFD group (p < 0.05). The most significant changes in serum glucose, total cholesterol, and triglyceride levels were confirmed in the GE40 group (p < 0.05). Epididymal fat was weighed and stained for body fat measurement, and significant differences were recorded from GE10 to GE40 (p < 0.05). Finally, 3T3-L1 pre-adipocytes were treated with GE, and cDNA from these cells was used for microarray analysis and qRT-PCR. Microarray analysis revealed 13 genes up-regulated and 21 genes down-regulated by GE. From the qRT-PCR analysis, we found that GE altered the mRNA expression of eosinophil peroxidase, glucose-dependent insulinotropic polypeptide receptor, and apolipoprotein B. Based on this study, we suggest that GR could be developed as an anti-obesity therapeutic agent.

Aquatic Exercise and Land Exercise Treatments after Total Knee Replacement Arthroplasty in Elderly Women: A Comparative Study.

Background and Objectives: Early intensive exercise after total knee replacement arthroplasty (TKRA) has become increasingly popular due to its ability to enhance knee physical function and reduce pain. When implemented exclusively, aquatic exercise (AE) appears to be more advantageous than land exercise (LE), particularly in the early phase after TKRA. Our study aimed to compare the clinical efficacy of AE and LE with respect to their effects on pain and physical function after TKRA. Materials and Methods: Between February 2008 and January 2020, 100 female patients who underwent TKRA were enrolled in this retrospective study. We measured the range of motion (ROM) of the knee, the isokinetic strength of the knee joint (function), and pain both initially and one month after TKRA. Two weeks after TKRA, the participants were enrolled in either the AE or the LE program for a total of two weeks. Two 30 min sessions of intensive ROM and knee strengthening exercises and balance training were provided to the AE and LE groups for 10 days. The home exercise group (HE) only received information on ROM and strengthening exercises. There were 33, 21, and 46 patients allocated to the AE, LE, and HE groups, respectively. Results: The ROM of the side on which surgery was performed improved significantly in all groups, as did the pain scores. In the AE group, the knee flexor strength showed a tendency toward improvement. Contrastingly, there was no significant improvement in the knee extensor strength in the AE group. Conclusions: Overall, the AE and LE groups showed superior outcomes compared with HE. In addition, the AE group demonstrated some improvement in knee muscle strength even with a short hospital stay. Further study with long-term follow-up should be performed to better define the outcomes.

Incidence and management of retrobulbar hemorrhage after blowout fracture repair.

BACKGROUND: Retrobulbar hemorrhage (RBH) is a rare complication after orbital surgery but associated with ocular complications including blindness. The aim of this study was to identify clinical characteristics of patients with RBH requiring emergent orbital decompression after blowout fracture repair. METHOD: A retrospective review of 426 blowout fracture patients at a tertiary oculoplastic clinic provided data regarding demographics, physical examination findings, and computed tomography (CT) images. Extraocular motility had been recorded in patient charts on a scale from 0 to - 4. Patients requiring emergent orbital decompression due to RBH after surgery (RBH group) were compared with those who did not (Control group), using the Mann-Whitney U-test. Incidences of RBH according to primary or secondary surgery were also investigated, using Fisher's exact test. RESULT: Five (1.2%) of the 426 patients who underwent blowout fracture repair developed RBH requiring emergent intervention. All RBH patients fully recovered after the decompression procedure or conservative treatment. Number of days to surgery was significantly longer in the RBH group (97.0 ± 80.1) than in the Control group (29.0 ± 253.0) (p = 0.05). Preoperative enophthalmos was also significantly greater in the RBH group (RBH vs. Control group, 3.6 ± 1.7 mm versus 1.2 ± 1.3 mm (p = 0.003)). The incidence of RBH was significantly higher in patients that underwent secondary surgery (odds ratio = 92.9 [95% confidence interval, 11.16-773.23], p = 0.001). CONCLUSIONS: Surgeons should pay more attention to hemostasis and postoperative care in patients with a large preoperative enophthalmic eye, when time from injury to surgery is long and in revision cases. When RBH occurs, time to intervention and surgical decompression is critical for visual recovery and preventing blindness. TRIAL REGISTRATION: The institutional review board of the Yeungnam University Medical Center approved this study ( YUMC 2018-11-010 ), which was conducted in accord with the Declaration of Helsinki.

Effect of Incorporating a Dimethacrylate Monomer on the Shear Bond Strength of Two Adhesive Resin Cements to Zirconia Ceramic.

Self-assembled nano-layering resulting from interaction of the phosphate functional group of adhesive monomers with zirconia ceramic surface has been proposed. The purpose of this study was to investigate the bond strengths of two adhesive resin cements (Panavia F 2.0 and BisCem) containing phosphate monomers added with various concentrations (0.0, 1.0, 3.0, and 5.0 wt%) of triethylene glycol dimethacrylate (TEGDMA) to air-abraded zirconia ceramic. The polished/air-abraded zirconia plates (KaVo Everest® ZS-Ronde) were imaged using atomic force microscopy and the average surface roughness (Ra) values were calculated (n = 5). The surface energy parameters of the zirconia plates and the resin cements were calculated based on the extended Fowkes theory. All resin-bonded (diameter: 2.38 mm) zirconia specimens were stored in water at 37 °C for 24 h and then half of them additionally thermocycled 10,000 times before the shear bond strength (SBS) test (n = 10). Air-abrasion of zirconia surface significantly increased the γhS (hydrogen bonding component) value (p < 0.001), as well as greatly increasing the surface area (p < 0.001). For both resin cements, the γhS (dipole-dipole component) gradually increased with increasing incorporated TEGDMA concentrations, whereas the γhS gradually decreased. Overall, the addition of 3.0 wt% of TEGDMA consistently resulted in higher SBS values even after thermocycling. Under the tested condition, reducing the concentration of the adhesive monomers with phosphate functional group by adding the dimethacrylate monomer (up to 3.0 wt%) increased the bond strength between the resin cements and zirconia ceramic.

Protein-induced metamorphosis of unilamellar lipid vesicles to multilamellar hybrid vesicles.

Protein encapsulation into nanocarriers has been extensively studied to improve the efficacy and stability of therapeutic proteins. However, the chemical modification of proteins or new synthetic carrier materials are essential to achieve a high encapsulation efficiency and structural stability of proteins, which hinders their clinical applications. New strategies to physically incorporate proteins into nanocarriers feasible for clinical uses are required to overcome the current limitation. Here we report the spontaneous protein-induced reorganization of 'pre-formed' unilamellar lipid vesicles to efficiently incorporate proteins within multilamellar protein-lipid hybrid vesicles without chemical modification. Epidermal growth factor (EGF) binds to the surface of cationic unilamellar lipid vesicles and induces layer-by-layer self-assembly of the vesicles. The protein is spontaneously entrapped in the interstitial layers of a multilamellar structure with extremely high loading efficiency, ~99%, through polyionic interactions as predicted by molecular dynamics simulation. The loaded protein exhibits much higher structural, chemical, and biological stability compared to free protein. The method is also successfully applied to several other proteins. This work provides a promising method for the highly efficient encapsulation of therapeutic proteins into multilamellar lipid vesicles without the use of specialized instruments, high energy, coupling agents, or organic solvents.

Self-Assembling Catalytic Peptide Nanomaterials Capable of Highly Efficient Peroxidase Activity.

The self-assembly of short peptides gives rise to versatile nanomaterials capable of promoting efficient catalysis. We have shown that short, seven-residue peptides bind hemin to produce functional catalytic materials which display highly efficient peroxidation activity, reaching a catalytic efficiency of 3×105 m-1 s-1 . Self-assembly is essential for catalysis as non-assembling controls show no activity. We have also observed peroxidase activity even in the absence of hemin, suggesting the potential to alter redox properties of substrates upon association with the assemblies. These results demonstrate the practical utility of self-assembled peptides in various catalytic applications and further support the evolutionary link between amyloids and modern-day enzymes.