Thermal Hysteresis and Bursting Rate in Sucrose Solutions with Antifreeze Proteins

Antifreeze proteins (AFPs) modify the ice shape and inhibit further growth leading to thermal hysteresis (TH). Numerous studies have been performed with the addtition of AFPs to preserve frozen products but the influence of sucrose on the effects of AFPs has not been investigated as of yet. Therefore, the TH activities of type I antifreeze protein (AFP I) and antifreeze glycoprotein (AFGP) were measured as a function of concentration, type of AFPs, and also sucrose concentration. The results showed that the TH values rose with increase in concentration of AFPs and sucrose concentration. The crystals experienced shape modification and grew in the c-axis direction in the presence of both AFPs. The bursting rate of crystals changed depending on both the concentrations of AFPs and sucrose.     

Cell-friendly regulation of ice crystals by antifreeze organism-inspired materials

There is a dilemma between ice growth inhibition and cell-threatening sharp-edged ice formation, dominating an unfavorable efficiency of cell cryoprotection using ice control materials (ICMs). Herein, we design different-type antifreeze organism-inspired ICMs and show the cell-friendly regulation of ice crystals by a synergistic effect for cell cryoprotection. Preferred rapid adsorption of “high-diffusivity ICM” on the curved ice crystal surface induced by “ice-binding ICM,” owing to the Gibbs–Curie–Wulff theorem, promotes smooth-shaped and small ice crystal formation during the freeze–thaw cycle. Meanwhile, their intracellular and extracellular osmotic balance well protects the cryopreserved cells. All-round protection benefits cell survival to achieve a cryopreservation efficiency of 97%. After cryopreservation, ice-binding ICM exhibits double thermosensitive behaviors to directly encapsulate post-thaw cells by gels, to further support cell-based therapies. This work provides an in-depth understanding of cell cryoprotection, and paves the road to the development of even better ICMs for biospecimen cryopreservation.     

Exploring novel cell cryoprotectants based on neutral amino acids

Cryoprotectants play a key role in cell cryopreservation because they can reduce cryoinjuries to cells associated with ice formation. To meet the clinical requirements of cryopreserved cells, cryoprotectants should be biocompatible, highly efficient and easily removable from cryopreserved cells. However, integration of these properties into one cryoprotectant still remains challenging. Herein, three biocompatible neutral amino acids, including β-alanine, γ-aminobutyric acid and ε-aminocaproic acid, are first reported to have the potential as such ideal cryoprotectants. The results demonstrate that they can inhibit ice formation and reduce osmotic stress to provide extracellular and intracellular protection, thereby ensuring high cryopreservation efficiency for both anuclear and nucleated cells. More importantly, due to the remarkable osmotic regulation ability, the neutral amino acids can be rapidly removed from cryopreserved cells via a one-step method without causing observable damage to cells, superior to the current state-of-the-art cryoprotectants—dimethyl sulfoxide and glycerol. This work provides a new perspective to develop novel cryoprotectants, which may have dramatic impacts on solvent-free cryopreservation technology to support the cell-based applications, such as cell therapy and tissue engineering, etc.     

An Effective Antifreeze Protein Predictor with Ensemble Classifiers and Comprehensive Sequence Descriptors

Antifreeze proteins (AFPs) play a pivotal role in the antifreeze effect of overwintering organisms. They have a wide range of applications in numerous fields, such as improving the production of crops and the quality of frozen foods. Accurate identification of AFPs may provide important clues to decipher the underlying mechanisms of AFPs in ice-binding and to facilitate the selection of the most appropriate AFPs for several applications. Based on an ensemble learning technique, this study proposes an AFP identification system called AFP-Ensemble. In this system, random forest classifiers are trained by different training subsets and then aggregated into a consensus classifier by majority voting. The resulting predictor yields a sensitivity of 0.892, a specificity of 0.940, an accuracy of 0.938 and a balanced accuracy of 0.916 on an independent dataset, which are far better than the results obtained by previous methods. These results reveal that AFP-Ensemble is an effective and promising predictor for large-scale determination of AFPs. The detailed feature analysis in this study may give useful insights into the molecular mechanisms of AFP-ice interactions and provide guidance for the related experimental validation. A web server has been designed to implement the proposed method.     

Growth control of ice crystals by poly(vinyl alcohol) and antifreeze protein in ice slurries

Effect of poly(vinyl alcohol) (PVA) in inhibiting an increase in ice crystal size in isothermal ice slurries was investigated, and then compared with the effect of an antifreeze protein (AFP), NaCl, and three other polymers, namely, poly(ethylene glycol), poly(vinyl pyrrolidone), and poly(acrylic acid). First, ice slurries, in which the initial size distribution of ice crystals was known, were isothermally preserved for given periods of time (typically 300 min) in the presence of PVA, AFP type I, NaCl, or the other three polymers. Then, the average size of the ice crystals was measured using image processing. Both the PVA and AFP type I completely inhibited the increase in ice crystal size at such low concentrations that the melting temperature of the solution was , whereas NaCl and the other three polymers clearly increased the ice crystal size due to Ostwald ripening. This inhibition effect of PVA and AFP type I was caused by thermal hysteresis, which is often taken as the primary manifestation of non-equilibrium antifreeze activity of these additives and defined as the difference between the melting temperature and non-equilibrium freezing temperature at which ice crystals start to grow in solution. The increase in ice crystal size was inhibited when the thermal hysteresis surpassed the driving potential for Ostwald ripening. Using PVA, which exhibits thermal hysteresis, is a novel technique to completely inhibit the increase in ice crystal size in isothermal ice slurries.     

Exploring novel cell cryoprotectants based on neutral amino acids

Cryoprotectants play a key role in cell cryopreservation because they can reduce cryoinjuries to cells associated with ice formation. To meet the clinical requirements of cryopreserved cells, cryoprotectants should be biocompatible, highly efficient and easily removable from cryopreserved cells. However, integration of these properties into one cryoprotectant still remains challenging. Herein, three biocompatible neutral amino acids, including β-alanine, γ-aminobutyric acid and ε-aminocaproic acid, are first reported to have the potential as such ideal cryoprotectants. The results demonstrate that they can inhibit ice formation and reduce osmotic stress to provide extracellular and intracellular protection, thereby ensuring high cryopreservation efficiency for both anuclear and nucleated cells. More importantly, due to the remarkable osmotic regulation ability, the neutral amino acids can be rapidly removed from cryopreserved cells via a one-step method without causing observable damage to cells, superior to the current state-of-the-art cryoprotectants—dimethyl sulfoxide and glycerol. This work provides a new perspective to develop novel cryoprotectants, which may have dramatic impacts on solvent-free cryopreservation technology to support the cell-based applications, such as cell therapy and tissue engineering, etc.     

Purification and partial characterization of antifreeze proteins from leaves of Ligustrum lucidum Ait

Three AFPs (AFP-1, AFP-2, AFP-3) of Ligustrum lucidum Ait leaves were purified by using ice-affinity, chromatography separation on a DEAE-cellulose-32 column and a Sephadex G100 column. The ice-affinity proteins were about 1.2 mg/100 g leaves. Their molecular weight was 66.1, 26.3, and 20.2 kDa, respectively. Their thermal hysteresis activity was 0.379, 0.678, and 0.460 °C respectively. Asx was very abundant in these AFPs; its molar ratio was 22.2, 24.9, and 27.8%, respectively. AFP-2 was effective to protect peroxidase, β-glucosidase, and trehalose synthase from freeze–thawing process. Cryoprotection of AFP-2 is better than trehalose.     

Tricine as a Novel Cryoprotectant with Osmotic Regulation,Ice Recrystallization Inhibition and Antioxidant Properties for Cryopreservation of Red Blood Cells

The cryopreservation of red blood cells (RBCs) plays a key role in blood transfusion therapy. Traditional cryoprotectants (CPAs) are mostly organic solvents and may cause side effects to RBCs, such as hemolysis and membrane damage. Therefore, it is necessary to find CPAs with a better performance and lower toxicity. Herein, we report for the first time that N-[Tri(hydroxymethyl)methyl]glycine (tricine) showed a great potential in the cryopreservation of sheep RBCs. The addition of tricine significantly increased the thawed RBCs’ recovery from 19.5 ± 1.8% to 81.2 ± 8.5%. The properties of thawed RBCs were also maintained normally. Through mathematical modeling analysis, tricine showed a great efficiency in cryopreservation. We found that tricine had a good osmotic regulation capacity, which could mitigate the dehydration of RBCs during cryopreservation. In addition, tricine inhibited ice recrystallization, thereby decreasing the mechanical damage from ice. Tricine could also reduce oxidative damage during freezing and thawing by scavenging reactive oxygen species (ROS) and maintaining the activities of endogenous antioxidant enzymes. This work is expected to open up a new path for the study of novel CPAs and promote the development of cryopreservation of RBCs.     

Using support vector machine and evolutionary profiles to predict antifreeze protein sequences

Antifreeze proteins (AFPs) are ice-binding proteins. Accurate identification of new AFPs is important in understanding ice-protein interactions and creating novel ice-binding domains in other proteins. In this paper, an accurate method, called AFP_PSSM, has been developed for predicting antifreeze proteins using a support vector machine (SVM) and position specific scoring matrix (PSSM) profiles. This is the first study in which evolutionary information in the form of PSSM profiles has been successfully used for predicting antifreeze proteins. Tested by 10-fold cross validation and independent test, the accuracy of the proposed method reaches 82.67% for the training dataset and 93.01% for the testing dataset, respectively. These results indicate that our predictor is a useful tool for predicting antifreeze proteins. A web server (AFP_PSSM) that implements the proposed predictor is freely available.     

Ice Crystal Growth in Sucrose Solutions Containing Kappa- and Iota-Carrageenans

Measurements of ice recrystallization inhibition (IRI) and thermal hysteresis (TH) activities of kappa (κ)- and iota (ι)-carrageenans were carried out to examine whether they can be novel cryoprotectants or not. IRI measurements indicate that both carrageenans reduce recrystallization in sucrose solution, but that the IRI activity of κ-carregeenan is higher than that of ι-carrageenan. TH measurements indicate that κ- and ι-carrageenans do not exhibit TH activity. TH activity measurements of antifreeze glycoprotein (AFGP) in the presence of κ-carregeenan demonstrate that this carregeenan neither influences the TH activity of AFGP nor the shape of the ice crystals. The round ice crystal shape transformed into an angular and elongated shape in the presence of both carregeenans.     

Cryopreservation of Mesenchymal Stem Cells Using Medical Grade Ice Nucleation Inducer

Mesenchymal stem cells (MSCs) can differentiate into multiple different tissue lineages and have favourable immunogenic potential making them an attractive prospect for regenerative medicine. As an essential part of the manufacturing process, preservation of these cells whilst maintaining potential is of critical importance. An uncontrolled area of storage remains the rate of change of temperature during freezing and thawing. Controlled-rate freezers attempted to rectify this; however, the change of phase from liquid to solid introduces two extreme phenomena; a rapid rise and a rapid fall in temperature in addition to the intended cooling rate (normally −1 °C/min) as a part of the supercooling event in cryopreservation. Nucleation events are well known to initiate the freezing transition although their active use in the form of ice nucleation devices (IND) are in their infancy in cryopreservation. This study sought to better understand the effects of ice nucleation and its active instigation with the use of an IND in both a standard cryotube with MSCs in suspension and a high-throughput adhered MSC 96-well plate set-up. A potential threshold nucleation temperature for best recovery of dental pulp MSCs may occur around −10 °C and for larger volume cell storage, IND and fast thaw creates the most stable process. For adhered cells, an IND with a slow thaw enables greatest metabolic activity post-thaw. This demonstrates a necessity for a medical grade IND to be used in future regenerative medicine manufacturing with the parameters discussed in this study to create stable products for clinical cellular therapies.     

聚乙二醇相变储能材料研究进展

聚乙二醇相变材料是一类相变焓较高、热滞后效应低的储能材料。本文综述了聚乙二醇相变储能材料的研究进展,重点论述了其制备方法、应用及展望。     

The thermal degradation of PET and analogous polyesters measured by thermal analysis-Fourier transform infrared spectroscopy

The thermal degradation of two commercial poly(ethylene terephthalate) (PET) samples and two laboratory prepared polyesters, poly(ethylene isophthalate) and poly(diethylene glycol terephthalate), was studied using thermogravimetry and thermal analysis-Fourier transform infrared spectroscopy. The commercial PET samples were copolymerised with diethylene glycol and isophthalic acid groups in different proportions, and their thermal stabilities were found to differ. Through a study of the thermal degradation of poly(diethylene glycol terephthalate) and poly(ethylene isophthalate), it was found that diethylene glycol and isophthalate units promoted thermal degradation through increased chain flexibility and more favourable bond angles, respectively. The thermal degradation of all the polyesters tested lead to the formation of non-volatile residue. Infrared spectroscopic analysis indicated that the residue consisted almost exclusively of interconnected aromatic rings.     

Preparation and characterization of poly(ethylene glycol) crosslinked chitosan films

Poly(ethylene glycol) (PEG) crosslinked chitosan films with various PEG to chitosan ratio and PEG molecular weight were successfully prepared via the epoxy‐amine reaction between chitosan and PEG‐epoxy. The thermal and mechanical properties and swelling behavior were studied for the PEG crosslinked chitosan films. The mechanical strength of chitosan films were greatly enforced by the introduction of PEG‐epoxy, achieving an elongation of about 80%. It was found that the crosslinked chitosan films form hydrogel in water, achieving a swelling ratio higher than 20 times of original weight. The swelling behavior of chitosan films relied greatly on the molecular weight of the crosslinker PEG‐epoxy and the weight percent of PEG‐epoxy. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 2008     

聚乙二醇/涤纶接枝共聚固-固相转变贮热材料

为了制备热稳定性好、蓄热性能优异的固-固相转变材料(PCM),研究采用化学法合成了聚乙二醇(PEG)/涤纶(PET)PCM。实验结果表明,PEG/PET PCM的热力学性能与PEG的分子量、PEG/PET质量配比以及不同交联体系有关。化学接枝法合成的PEG/PET PCM,最大相变焓可达112.02 J/g,热稳定性提高,热滞后性减小,PEG/PET PCM在众多领域具有广泛应用。     

ABA triblock copolymers from poly(p‐dioxanone) and poly(ethylene glycol)

Poly(p‐dioxanone)–poly(ethylene glycol)–poly(p‐dioxanone) ABA triblock copolymers (PEDO) were synthesized by ring‐opening polymerization from p‐dioxanone using poly(ethylene glycol) (PEG) with different molecular weights as macroinitiators in N₂ atmosphere. The copolymer was characterized by ¹H NMR spectroscope. The thermal behavior, crystallization, and thermal stability of these copolymers were investigated by differential scanning calorimetry and thermogravimetric measurements. The water absorption of these copolymers was also measured. The results indicated that the content and length of PEG chain have a greater effect on the properties of copolymers. This kind of biodegradable copolymer will find a potential application in biomedical materials. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 102:1092–1097, 2006     

导电聚合物PEDOT/PSS-MPEG的制备及性能

引言导电高分子既有导体材料的光电学特性,又有良好的力学性能和可加工性[1],这使得导电高分子材料具有广泛的应用前景。聚噻吩类有机导电材料[2]就是这类材料中的一种。聚噻吩的室温电导率     

Blood-Based Biomarkers in Hepatitis B Virus-Related Hepatocellular Carcinoma,Including the Viral Genome and Glycosylated Proteins

Hepatitis B virus (HBV) infection is a major risk factor for hepatocellular carcinoma (HCC) development and is a global public health issue. High performance biomarkers can aid the early detection of HCC development in HBV-infected individuals. In addition, advances in the understanding of the pathogenesis of HBV infection and in clinical laboratory techniques have enabled the establishment of disease-specific tests, prediction of the progression of liver diseases, including HCC, and auxiliary diagnosis of HCC, using blood-based methods instead of biopsies of liver or HCC tissues. Viral factors such as the HBV genotype, HBV genetic mutations, HBV DNA, and HBV-related antigens, as well as host factors, such as tumor-associated proteins and post-translational modifications, especially glycosylated proteins, can be blood-based, disease-specific biomarkers for HCC development in HBV-infected patients. In this review, we describe the clinical applications of viral biomarkers, including the HBV genome and glycosylated proteins, for patients at a risk of HBV-related HCC, based on their molecular mechanisms. In addition, we introduce promising biomarker candidates for practical use, including colony stimulating factor 1 receptor (CSF1R), extracellular vesicles, and cell-free, circulating tumor DNA. The clinical use of such surrogate markers may lead to a better understanding of the risk of disease progression and early detection of HCC in HBV-infected patients, thereby improving their prognosis.     

The Role of PEMFs on Bone Healing: An In Vitro Study

Bone responses to pulsed electromagnetic fields (PEMFs) have been extensively studied by using devices that expose bone cells to PEMFs to stimulate extracellular matrix (ECM) synthesis for bone and cartilage repair. The aim of this work was to highlight in which bone healing phase PEMFs exert their action. Specifically, we evaluated the effects of PEMFs both on human adipose mesenchymal stem cells (hASCs) and on primary human osteoblasts (hOBs) by testing gene and protein expression of early bone markers (on hASCs) and the synthesis of late bone-specific proteins (on hOBs) as markers of bone remodeling. Our results indicate that PEMFs seem to exert their action on bone formation, acting on osteogenic precursors (hASCs) and inducing the commitment towards the differentiation pathways, unlike mature and terminally differentiated cells (hOBs), which are known to resist homeostasis perturbation more and seem to be much less responsive than mesenchymal stem cells. Understanding the role of PEMFs on bone regenerative processes provides important details for their clinical application.