Hepatocellular Calcification in Severe Ischemia–Reperfusion Injury in a Liver Allograft

Ischemia–reperfusion injury (IR) of solid organ allografts is a consequence of ischemia resulting from disruption of blood flow during organ harvest and transportation. Histologically, this manifests as variable necrosis in a pattern similar to that seen in systemic hypoperfusion. Calcification of hepatocytes has been rarely observed in ischemic injury due to systemic shock and in two cases of severe IR, both of which were associated with graft loss and death. The authors present another case of dystrophic calcification within hepatocytes occurring in a liver allograft affected by severe IR. Biochemical stains revealed that the mineralized material was calcium phosphate (likely hydroxyapatite). By electron microscopy, the hepatocyte cytoplasm was filled with variably calcified vacuoles, a subset of which likely represented swollen mitochondria. When encountered in hepatic allograft biopsies, hepatocellular calcification is associated with ischemic injury and a poor prognosis.     

一种基于FPGA的数字EIT系统的实验研究

以FPGA芯片为核心的数字化生物电阻抗测量实验平台,主要完成直接数字频率合成(DDS)的电流源模块、V/I变换和数字解调模块的研究.采用DDS技术生成正弦信号发生器,由THS4021改进的Howland电路设计压控电流源,在不同负载条件下测量其输出电流幅值的频率响应,并在不同频率下测量电流源的输出阻抗,对恒流源性能的稳定性和精度进行测定.解调方法采用数字化解调,测试不同激励频率下的数字正交解调输出误差变化情况.最后采用盐水槽实验系统进行成像实验.结果表明,电流源可在6.1～390.6 kHz范围输出多频激励信号,在600 kHz前保持在190 kΩ的输出阻抗.当信号频率从200 kHz逐渐增加到1.6 MHz时,解调电路输出的I和Q路误差逐渐增加,对应的幅度误差从1.13％增加到7.19％,相位误差从1.03％增大到5.34％.采用相邻激励-相邻测量模式对环氧树脂棒进行盐水槽成像实验,成像结果表明系统能够对单个目标物体实现较准确的定位,验证了本研究平台的可行性.     

Functionalized liposomes loaded with siRNAs targeting ion channels in effector memory T cells as a potential therapy for autoimmunity

Effector memory T cells (T_M) play a key role in the pathology of certain autoimmune disorders. The activity of effector T_M cells is under the control of Kv1.3 ion channels, which facilitate the Ca²⁺ influx necessary for T cell activation and function, i.e. cytokine release and proliferation. Consequently, the knock-down of Kv1.3 expression in effector T_M's may be utilized as a therapy for the treatment of autoimmune diseases. In this study we synthesized lipid unilamellar nanoparticles (NPs) that can selectively deliver Kv1.3 siRNAs into T_M cells in vitro. NPs made from a mixture of phosphatidylcholine, pegylated/biotinylated phosphoethanolamine and cholesterol were functionalized with biotinylated-CD45RO (cell surface marker of T_M's) antibodies via fluorophore-conjugated streptavidin (CD45RO-NPs). Incubation of T cells with CD45RO-NPs resulted into the selective attachment and endocytosis of the NPs into T_M's. Furthermore, the siRNA against Kv1.3, encapsulated into the CD45RO-NPs, was released into the cytosol. Consequently, the expression of Kv1.3 channels decreased significantly in T_M's, which led to a remarkable decrease in Ca²⁺ influx. Our results can form the basis of an innovative therapeutic approach in autoimmunity.     

Identification of a Suppressor of Retinal Degeneration in Drosophila Photoreceptors

During sensory transduction, Drosophila photoreceptors experience substantial increases in intracellular Ca²⁺ levels ([Ca²⁺]_i). Nevertheless in a number of mutants associated with excessive Ca²⁺ influx through transient receptor potential (TRP) channels, Drosophila photoreceptors undergo loss of normal cellular structure manifest as a retinal degeneration. However, the molecular mechanisms that underpin this degeneration process remain unclear. The authors previously isolated a mutant, su(40), that is able to suppress the retinal degeneration seen in photoreceptors from loss-of-function alleles of rdgA that are known to have constitutively active TRP channels. Here the authors report the genetic mapping of su(40) as well the isolation of additional alleles of su(40). Studies of su(40) as well as these new alleles should facilitate the understanding of the mechanisms by which excessive Ca²⁺ influx results in retinal degeneration.     

Enamel Mineralization and the Role of Ameloblasts in Calcium Transport

Amelogenesis is a dynamic and unique process of cell-matrix interactions in that matrix synthesis, degradation and resorption all proceed simultaneously, coupled with mineral depositions in a compartment between ameloblasts and dentin or dental papilla. Accumulation of data suggest the role of ameloblasts in tooth morphogenesis and matrix formation, but no fully acceptable explanation has been given concerning the role of ameloblasts in calcium transport. In this article, old and new points of issue raised regarding the role of ameloblasts in calcium acquisition are reviewed and possible mechanisms whereby the ameloblasts prevent the rise of cytosolic calcium while actively or less actively transporting calcium are elaborated upon based on recent findings.     

Novel light-curable materials containing experimental bioactive micro-fillers remineralise mineral-depleted bonded-dentine interfaces

This study aimed at evaluating the therapeutic remineralising effects of innovative light-curable materials (LCMs) containing two experimental calcium silicate-based micro-fillers (TCS) modified with β-TCP only or β-TCP, zinc oxide (ZnO)/polyacrylic acid (PAA) on mineral-depleted bonded-dentine interfaces in simulated body fluids (SBFS). Three experimental LCMs were formulated: (1) resin A, containing a β-TCP-modified TCS (βTCS) micro-filler; (2) resin B, containing a polycarboxylated β-TCP/ZnO-modified TCS (βZn-TCS) micro-filler; and 3) resin C, containing no filler (control). Acid-etched (35% H₃PO₄) dentine specimens were bonded using the three LCMs and submitted to atomic force microscope (AFM)/nano-indentation analysis to evaluate the modulus of elasticity (Ei) and hardness (Hi) across the interface after SBFS storage (24?h/1?m/3?m). The ultramorphology and micropermeability of the resin–dentine interface were evaluated using confocal laser microscopy. Resin–dentine sticks were created and submitted to microtensile bond strength (μTBS) test (SBFS: 24?h/3?m). Scanning electron microscopy (SEM) was performed after de-bonding. The LCMs containing the experimental bioactive micro-fillers reduced the micropermeability and induced a significant increase of the Ei and Hi along the bonding interface. The specimens created using the resin B (βZn-TCS) attained the highest μTBS values both after 24?h and 3?m of SBFS storage. In conclusion, the innovative bioactive light-curable materials tested in this study are able to induce a therapeutic remineralising effect on the nano-mechanical properties and on the sealing ability of mineral-depleted resin–dentine interfaces. The contemporary idea of minimally invasive operative treatment, where therapeutic restorations are performed to combat the carious process and remineralise the dental hard tissues, may be satisfied by using such resin-base systems, containing βTCS or βZn-TCS bioactive micro-fillers.     

Biocompatibility evaluation of emulsion electrospun nanofibers using osteoblasts for bone tissue engineering

Emulsion electrospinning is an advanced technique to fabricate core-shell structured nanofibrous scaffolds, with great potential for drug encapsulation. Incorporation of dual factors hydroxyapatite (HA) and laminin, respectively, within the shell and core of nanofibers through emulsion electrospinning might be of advantageous in supporting the adhesion, proliferation, and maturation of cells instead of single factor-encapsulated nanofibers. We fabricated poly(L-lactic acid-co-?-caprolactone) (PLCL)/hydroxyapaptite (PLCL/HA), PLCL/laminin (PLCL/Lam), and PLCL/hydroxyapatite/laminin (PLCL/HA/Lam) scaffolds with fiber diameter of 388?±?35, 388?±?81, and 379?±?57?nm, respectively, by emulsion electrospinning. The elastic modulus of the prepared scaffolds ranged from 22.7–37.0?MPa. The osteoblast proliferation on PLCL/HA/Lam scaffolds, determined on day 21, was found 10.4% and 12.0% higher than the cell proliferation on PLCL/Lam or PLCL/HA scaffold, respectively. Cell maturation determined on day 14, by alkaline phosphatase (ALP) activity, was significantly higher on PLCL/HA/Lam scaffolds than the ALP activity on PLCL/HA and PLCL/Lam scaffolds (p???0.05). Results of the energy dispersive X-ray studies carried out on day 28 also showed higher calcium deposition by cells seeded on PLCL/HA/Lam scaffolds. Osteoblasts were found to adhere, proliferate, and mature actively on PLCL/HA/Lam nanofibers with enhanced cell proliferation, ALP activity, bone protein expression, and mineral deposition. Based on the results, we can conclude that laminin and HA individually played roles in osteoblast proliferation and maturation, and the synergistic function of both factors within the novel emulsion electrospun PLCL/HA/Lam nanofibers enhanced the functionality of osteoblasts, confirming their potential application in bone tissue regeneration.