Tendon stem/progenitor cell ageing: Modulation and rejuvenation

Tendon ageing is a complicated process caused by multifaceted pathways and ageing plays a critical role in the occurrence and severity of tendon injury. The role of tendon stem/progenitor cells (TSPCs) in tendon maintenance and regeneration has received increasing attention in recent years. The decreased capacity of TSPCs in seniors contributes to impaired tendon functions and raises questions as to what extent these cells either affect, or cause ageing, and whether these age-related cellular alterations are caused by intrinsic factors or the cellular environment. In this review, recent discoveries concerning the biological characteristics of TSPCs and age-related changes in TSPCs, including the effects of cellular epigenetic alterations and the mechanisms involved in the ageing process, are analyzed. During the ageing process, TSPCs ageing might occur as a natural part of the tendon ageing, but could also result from decreased levels of growth factor, hormone deficits and changes in other related factors. Here, we discuss methods that might induce the rejuvenation of TSPC functions that are impaired during ageing, including moderate exercise, cell extracellular matrix condition, growth factors and hormones; these methods aim to rejuvenate the features of youthfulness with the ultimate goal of improving human health during ageing.     

The gamma-chain cytokine/receptor system in fish: more ligands and receptors

The mammalian gamma-chain (γC) cytokine family consists of interleukin (IL)-2, IL-4, IL-7, IL-9, IL-15 and IL-21. They signal through a receptor complex containing the common γC and a private alpha chain, and in the case of IL-2 and IL-15 an additional common IL-2/15Rβ chain. Deficiency of γC signalling in mammals prevents CD4⁺ T cells from developing effector functions and CD8⁺ T cells from developing immunological memory. Thus γC cytokines are critical for the generation and peripheral homeostasis of naïve and memory T cells. This review will give an update on the γC ligands and receptor subunits in fish, and also present some new data on the cloning and expression of a second γC and two IL-2Rβ chains in rainbow trout Oncorhynchus mykiss. In recent years, aided by the availability of sequenced fish genomes and expressed sequence tag databases, five of the six mammalian γC cytokines and their cognate receptors have been discovered in fish, with only the IL-9/IL-9R homologues apparently absent. Paralogues have been discovered in diploid fish and all the receptors described in the tetraploid rainbow trout, including γC itself, IL-2Rβ, IL-4Rα, IL-13Rα1, IL-13Rα2 and IL-2/15Rα, have duplicates. As a consequence of the teleost and salmonid whole genome duplications, even more paralogues may yet be discovered. Some of the paralogues have changes in domain structures and show differential expression and modulation, suggesting the potential for a change in function. Functional characterisation of fish γC cytokines is beginning but made more difficult by the co-existence of so many paralogues of the ligands and their receptors. Initial functional studies have shown that fish γC cytokines can modulate the expression of key cytokines (e.g. interferon-γ, IL-10 and IL-22) of the adaptive immune response, and may thus have promise as adjuvants to improve vaccination efficiency in fish.     

细胞外调控分子对酸敏感离子通道的功能调控及其机制

酸敏感离子通道(acid-Sensing ion channels,ASlCs)是一类由细胞外质子(H )激活的配体门控阳离子通道.迄今为止,人们在哺乳动物体内已经发现了6种ASICs亚基蛋白,它们分布在多种组织器官中.越来越多的研究表明:ASICs参与了机体的生理、病理过程,如:学习、记忆、痛觉、脑中风和肿瘤.在过去的10年中,人们发现多种内源性或外源性分子可以调控ASICs通道活性.由于这些细胞外调控分子与多种生理和病理功能有关,因此研究细胞外调控分子对ASICs的调控及其分子机制,可以帮助我们更多地了解ASICs功能以及结构信息,也为人们设计ASICs靶点特异性药物提供了理论依据.文章将系统地介绍细胞外调控分子对ASICs的功能调控及其作用机制,特别是该研究领域的最新进展.     

Phosphate Sorption and Desorption on Pyrite in Primitive Aqueous Scenarios: Relevance of acidic → Alkaline Transitions

Phosphate (P _i) sorption assays onto pyrite in media simulating primeval aquatic scenarios affected by hydrothermal emissions, reveal that acidic conditions favour P _i sorption whereas mild alkaline media – as well as those simulating sulfur oxidation to SO²⁻ ₄ – revert this capture process. Several mechanisms relevant to P _i availability in prebiotic eras are implicated in the modulation of these processes. Those favouring sorption are: (a) hydrophobic coating of molecules, such as acetate that could be formed in the vicinity of hydrothermal vents; (b) water and Mg²⁺ bridging in the interface mineral-aqueous media; (c) surface charge neutralization by monovalent cations (Na⁺ and K⁺). The increase of both the medium pH and the SO²⁻ ₄ trapping by the mineral interface would provoke the release of sorbed P _i due to charge polarization. Moreover it is shown that P _i self-modulates its sorption, a mechanism that depends on the abundance of SO²⁻ ₄ in the interface. The relevance of the proposed mechanisms of P _i capture, release and trapping arises from the need of abundant presence of this molecule for primitive phosphorylations, since – similarly to contemporary aqueous media – inorganic phosphate concentrations in primitive seas should have been low. It is proposed that the presence of sulphide minerals with high affinity to P _i could have trapped this molecule in an efficient manner, allowing its concentration in specific niches. In these niches, the conditions studied in the present work would have been relevant for its availability in soluble form, specially in primitive insulated systems with pH gradients across the wall. R B-L and Y C-S contributed equally to this work; recipients of fellowships from the Brazilian National Research Council in the PIBIC and PINC-School of Medicine programs of the Universidade Federal de Rio de Janeiro     

Entrainment of Na/K pumps by a synchronization modulation electric field

We studied entrainment of the catalytic cycle of the Na/K pumps by an imposed external AC electric field. Our results show that a well designed dichotomous oscillating electric field with a frequency close to the pumps’ natural turnover rate can synchronize the pump molecules. Characteristics of the synchronized pumps include: (1) outward pump currents responding to Na-extrusion and inward pump currents responding to K-pumping in are separated; (2) magnitude of the outward pump currents can be up to three times higher than that of the randomly paced pump currents; (3) magnitude ratio of the outward over inward pump currents reveals the 3:2 stoichiometry of the pumps. We, further, gradually increased the field oscillating frequency in a stepwise pattern and kept pump synchronization in each step. We found that the pumps’ turnover rate could be modulated up as the field frequency increased. Consequently, the pump currents significantly increased by many fold. In summary, these results show that the catalytic cycle of Na/K pumps can be synchronized and modulated by a well designed oscillating electric field resulting in activation of the pump functions.     

Presynaptic kainate receptor-mediated bidirectional modulatory actions: Mechanisms

Kainate receptors (KARs) are members of the glutamate receptor family, which also includes two other ionotropic subtypes, i.e. NMDA- and AMPA-type receptors, and types I, II and III metabotropic glutamate receptors. KARs mediate synaptic transmission postynaptically through their ionotropic capacity, while presynaptically, they modulate the release of both GABA and glutamate through operationally diverse modus operandi. At hippocampal mossy fiber (MF)-CA3 synapses, KARs have a biphasic effect on glutamate release, such that, depending on the extent of their activation, a facilitation or depression of glutamate release can be observed. This modulation is posited to contribute to important roles of KARs in short- and long-term plasticity. Elucidation of the modes of action of KARs in their depression and facilitation of glutamate release is beginning to gather impetus. Here we will focus on the cellular mechanisms involved in the modulation of glutamate release by presynaptic KAR activation at MF-CA3 synapses, a field that has seen significant progress in recent years.     

Enhancing the functional properties of thermophilic enzymes by chemical modification and immobilization

The immobilization of proteins (mostly typically enzymes) onto solid supports is mature technology and has been used successfully to enhance biocatalytic processes in a wide range of industrial applications. However, continued developments in immobilization technology have led to more sophisticated and specialized applications of the process. A combination of targeted chemistries, for both the support and the protein, sometimes in combination with additional chemical and/or genetic engineering, has led to the development of methods for the modification of protein functional properties, for enhancing protein stability and for the recovery of specific proteins from complex mixtures. In particular, the development of effective methods for immobilizing large multi-subunit proteins with multiple covalent linkages (multi-point immobilization) has been effective in stabilizing proteins where subunit dissociation is the initial step in enzyme inactivation. In some instances, multiple benefits are achievable in a single process.Here we comprehensively review the literature pertaining to immobilization and chemical modification of different enzyme classes from thermophiles, with emphasis on the chemistries involved and their implications for modification of the enzyme functional properties. We also highlight the potential for synergies in the combined use of immobilization and other chemical modifications.     

Modulation of a lipase from Staphylococcus warneri EX17 using immobilization techniques

This research describes the immobilization on glyoxyl, cyanogen bromide or octyl agarose beads of a purified lipase from Staphylococcus warneri strain EX17 (SWL), and the effect on its properties. The immobilization on glyoxyl-agarose at pH 10 and 25 °C, conditions in which the enzyme is readily inactivated, required the stabilization of the soluble enzyme. This was attained by the addition of 25% glycerol. Using this additive, immobilization on glyoxyl-agarose beads proceeded very quickly with good activity retention around 80%. This was the most stable preparation under thermal inactivation at pH 5, 7 and 9, in the presence of either cosolvents or detergents. This preparation was hyperactivated by concentrations of Triton X-100, which would produce negative effects over enzyme activity when using the other SWL preparations. Immobilized SWL preparations hydrolyzed different chiral esters, such as (±)-methyl mandelate, (±)-2-O-butyryl-2-phenylacetic acid, and (±)-2-hydroxy-4-phenyl-butyric acid ethyl ester, being its specificity depended on the immobilization protocol. The enantiospecificity was also strongly modulated by the immobilization. Thus, using HPBEt as substrate, octyl-SWL exhibited an opposite enantiospecificity to the other two biocatalysts. This preparation was the most enantioselective in the hydrolysis of (±)-2-O-butyryl-2-phenylacetic acid (E = 56.3).     

大豆基因工程根瘤菌田间结瘤和共生固氮效应

利用三亲本接合转移方法,将快生型大豆根病菌B_(52)的基因文库克隆转移到受体菌慢生型大豆根瘤菌2210中,获得4株大豆基因工程根瘤菌(32.43、B—2—6、B—3—8)。田间接种试验表明:在大豆分枝期(V2),43和32结瘤效分别比不接种对照增加117.9％和100.4％;在初花期(R_1),32和B—2—6结瘤数分别比对照增加122.5％和91.6％,根瘤固氮酶活性增加233.3％和195.6％;在结荚始期(R3),B—2—6和32结瘤数比对照增加78.5％和70.1％,根瘤固氟酶活性43和B—2—6分别比对照增加53.4％和49.3％。产量统计表明:32、B—2—6和43比对照分别增产16.8％、14.3％和12.2％,亩增收大豆分别为28.2、24.1和20.6公斤。以上三个菌株均比受体菌株2210增产率高。     

Ganglioside-Modulated Proteolysis by Ca2+-Activated Neutral Proteinase (CANP): A Role of Glycoconjugates in CANP Regulation

We examined ganglioside modulation of the activity of the millimolar Ca2(+)-sensitive form (mCANP) of calcium-activated neutral proteinase (CANP), which is enriched in myelin, from brain. GM1, GD1a, GT1a, GM2, and GM4 produced a concentration-dependent increase of mCANP activity. GD1a stimulated the greatest increase of enzyme activity (107%), followed by GT1a, whereas GD1b was inhibitory (56%). GM1, GM2, and GM4 stimulated but less so than GD1a and GT1a. Free N-acetylneuraminic acid, asialo-GM1, GM3, and a ganglioside mixture containing GM1, GD3, GD1a, and GD1b had no effect. The ganglioside-mediated modulation was not affected by trifluoperazine and chlorpromazine (phospholipid-binding antagonists). The mCANP Ca2+ requirement was significantly reduced in the presence of stimulatory gangliosides, and this increased sensitivity varied (10-50-fold) with ganglioside structure. Gangliosides may interact with membrane mCANP and modulate its proteolytic action.