Exogenous Expression of Human apoA-I Enhances Cardiac Differentiation of Pluripotent Stem Cells

The cardioprotective effects of high-density lipoprotein cholesterol (HDL-C) and apolipoprotein A1 (apoA-I) are well documented, but their effects in the direction of the cardiac differentiation of embryonic stem cells are unknown. We evaluated the effects of exogenous apoA-I expression on cardiac differentiation of ESCs and maturation of ESC-derived cardiomyocytes. We stably over-expressed full-length human apoA-I cDNA with lentivirus (LV)-mediated gene transfer in undifferentiated mouse ESCs and human induced pluripotent stem cells. Upon cardiac differentiation, we observed a significantly higher percentage of beating embryoid bodies, an increased number of cardiomyocytes as determined by flow cytometry, and expression of cardiac markers including α-myosin heavy chain, β-myosin heavy chain and myosin light chain 2 ventricular transcripts in LV-apoA-I transduced ESCs compared with control (LV-GFP). In the presence of noggin, a BMP4 antagonist, activation of BMP4-SMAD signaling cascade in apoA-I transduced ESCs completely abolished the apoA-I stimulated cardiac differentiation. Furthermore, co-application of recombinant apoA-I and BMP4 synergistically increased the percentage of beating EBs derived from untransduced D3 ESCs. These together suggests that that pro-cardiogenic apoA-I is mediated via the BMP4-SMAD signaling pathway. Functionally, cardiomyocytes derived from the apoA-I-transduced cells exhibited improved calcium handling properties in both non-caffeine and caffeine-induced calcium transient, suggesting that apoA-I plays a role in enhancing cardiac maturation. This increased cardiac differentiation and maturation has also been observed in human iPSCs, providing further evidence of the beneficial effects of apoA-I in promoting cardiac differentiation. In Conclusion, we present novel experimental evidence that apoA-I enhances cardiac differentiation of ESCs and iPSCs and promotes maturation of the calcium handling property of ESC-derived cardiomyocytes via the BMP4/SMAD signaling pathway

The effects of a mindfulness-based family psychoeducation intervention for the caregivers of young adults with first-episode psychosis: A randomized controlled trial

Objective: In this study, we investigated the effects of a mindfulness-based family psychoeducation (MBFPE) program on the mental-health outcomes of both caregivers and young adults with first-episode psychosis with an onset in the past three years through a multi-site randomized controlled trial. We also studied the outcomes of three potential mediating effects of interpersonal mindfulness, expressed emotions, and non-attachment on the program. Method: We randomly assigned 65 caregivers of young adults with psychosis to MBFPE (n = 33) or an ordinary family psychoeducation (FPE) program (n = 32); among them, 18 young adults in recovery also participated in the evaluation of outcomes. Results: Intent-to-treat analyses were conducted. No significant time × group interaction effects of MBFPE and FPE programs were found in any of the caregivers’ outcomes. However, the young adults with psychosis reported higher levels of recovery after the MBFPE program than after the ordinary FPE program (F = 8.268, p = 0.012, d = 1.484). They also reported a larger reduction in over-involvement of their caregivers (F = 4.846, p = 0.044, d = 1.136), showing that MBFPE had a superior effect to FPE in promoting recovery and reducing over-involvement. Conclusions: A brief psychoeducation program may not reduce the burden on or improve the mental-health outcome of caregivers of individuals with recent-onset psychosis. However, integrating mindfulness into a conventional family psychoeducation program may reduce the expressed emotions of caregivers, especially over-involvement. Further studies should explore how psychoeducation programs can reduce the impact of psychosis on family through sustainable effects in terms of reducing their burden and expressed emotions, using a rigorous study and adequate sample size

BING, a novel antimicrobial peptide isolated from Japanese medaka plasma, targets bacterial envelope stress response by suppressing cpxR expression

Antimicrobial peptides (AMPs) have emerged as a promising alternative to small molecule antibiotics. Although AMPs have previously been isolated in many organisms, efforts on the systematic identification of AMPs in fish have been lagging. Here, we collected peptides from the plasma of medaka (Oryzias latipes) fish. By using mass spectrometry, 6399 unique sequences were identified from the isolated peptides, among which 430 peptides were bioinformatically predicted to be potential AMPs. One of them, a thermostable 13-residue peptide named BING, shows a broad-spectrum toxicity against pathogenic bacteria including drug-resistant strains, at concentrations that presented relatively low toxicity to mammalian cell lines and medaka. Proteomic analysis indicated that BING treatment induced a deregulation of periplasmic peptidyl-prolyl isomerases in gram-negative bacteria. We observed that BING reduced the RNA level of cpxR, an upstream regulator of envelope stress responses. cpxR is known to play a crucial role in the development of antimicrobial resistance, including the regulation of genes involved in drug efflux. BING downregulated the expression of efflux pump components mexB, mexY and oprM in P. aeruginosa and significantly synergised the toxicity of antibiotics towards these bacteria. In addition, exposure to sublethal doses of BING delayed the development of antibiotic resistance. To our knowledge, BING is the first AMP shown to suppress cpxR expression in Gram-negative bacteria. This discovery highlights the cpxR pathway as a potential antimicrobial target

Calcium Homeostasis in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Rationale: Cardiomyocytes generated from human induced pluripotent stem cells (hiPSCs) are suggested as the most promising candidate to replenish cardiomyocyte loss in regenerative medicine. Little is known about their calcium homeostasis, the key process underlying excitation-contraction coupling. Objective: We investigated the calcium handling properties of hiPSC-derived cardiomyocytes and compared with those from human embryonic stem cells (hESCs). Methods and Results: We differentiated cardiomyocytes from hiPSCs (IMR90 and KS1) and hESCs (H7 and HES3) with established protocols. Beating outgrowths from embryoid bodies were typically observed 2 weeks after induction. Cells in these outgrowths were stained positively for tropomyosin and sarcomeric alpha-actinin. Reverse-transcription polymerase chain reaction studies demonstrated the expressions of cardiac-specific markers in both hiPSC- and hESC-derived cardiomyocytes. Calcium handling properties of 20-day-old hiPSC- and hESC-derived cardiomyocytes were investigated using fluorescence confocal microscopy. Compared with hESC-derived cardiomyocytes, spontaneous calcium transients from both lines of hiPSC-derived cardiomyocytes were of significantly smaller amplitude and with slower maximal upstroke velocity. Better caffeine-induced calcium handling kinetics in hESC-CMs indicates a higher sacroplasmic recticulum calcium store. Furthermore, in contrast with hESC-derived cardiomyocytes, ryanodine did not reduce the amplitudes, maximal upstroke and decay velocity of calcium transients of hiPSC-derived cardiomyocytes. In addition, spatial inhomogeneity in temporal properties of calcium transients across the width of cardiomyocytes was more pronounced in hiPSC-derived cardiomyocytes than their hESC counterpart as revealed line-scan calcium imaging. Expressions of the key calcium-handling proteins including ryanodine recptor-2 (RyR2), sacroplasmic recticulum calcium-ATPase (SERCA), junction (Jun) and triadin (TRDN), were significantly lower in hiPSC than in hESCs. Conclusions: The results indicate the calcium handling properties of hiPSC-derived cardiomyocytes are relatively immature to hESC counterparts. © 2011 The Author(s).published_or_final_versionSpringer Open Choice, 21 Feb 201

死亡與臨終 : 事實與迷思 = Death and dying : myth and reality

我們都必定會經歷臨終與死亡，但我們卻不可能在死亡來臨之前了解它是怎麼一回事。 醫療進步與文化變遷使大衆與臨終和死亡隔離。隨着醫療進步而來的，就是以防止死亡為醫療目標，因而死亡變成一項醫療上的失敗。 在現代社會，死亡走向制度化與非人化。臨終過程由醫護人員負責處理，於是對很多人來說，這過程變得很神秘。恐懼與焦慮與其說由於對死亡的認知引起，不如說由於其不可知而導致。對死亡與臨終的了解雖然不一定消除恐懼與焦慮，但肯定是對不可知的恐懼的唯一對治之方

Advance care planning for 600 Chinese patients with end-stage renal disease

Background/purpose: There is increasing recognition of the need to integrate advance care planning (ACP) into end-stage renal disease (ESRD) care with attention to medical, ethical, psychosocial, and spiritual issues but publications comparing patients who chose renal replacement therapy (RRT) and renal palliative care (RPC) is scarce. We here share our experience on ACP for ESRD patients in a center with renal replacement and palliative programs in place. Methods: From June 2006 to December 2011, ESRD patients were empowered to make an informed choice of future medical care in a structured ACP that was emphasized to be an ongoing process. Patients who opted for RRT and RPC would be followed up at the predialysis clinic and the one-stop multidisciplinary RPC clinic, respectively. This was a single-center study in a secondary care hospital. A total of 600 patients (265 RRT, 335 RPC) were enrolled and followed up over a median of 782 days. Results: The majority of patients and relatives declined dialysis because of perceived physical burden. Only 1.6% of palliative care patients changed their decision and commenced dialysis. Baseline characteristics differed between patients who chose RRT or RPC. Survival declined according to the modified Charlson Comorbidity Index scores. Older age, mental incompetence, hyperlipidemia, high modified Charlson Comorbidity Index, low estimated glomerular filtration rate, and low albumin were important independent predictors of poor survival. Factors affecting the ACP decision were discussed in the Chinese culture context. Conclusion: A structured ACP could empower the patient to make an informed decision on the management of ESRD. 背景: 於未期腎病患者的照顧中加入關注身心社靈和倫理問題的預設照顧計劃(ACP)受到日益重視，但有關比較接受腎替代療法和接受腎臟紓緩治療文獻討論為數不多。作為同時提供腎透析服務以及腎臟紓緩治療的部門，本文旨在分享我們為未期腎病者討論預計照顧計劃的經驗。 方法: 自二零零六年六月至二零一零五月間，透過有組織的預設照顧計劃討論，未期腎病患者會被鼓勵就未來的治療計劃作出知情選擇。選擇腎透析和腎臟紓緩治療的病人會分別於透析預備門診和一站式跨科際腎臟紓緩治療門診去覆診。本研究於一家二級醫院進行。總共有六百病人參與此研究，當中265名接受腎透析，335名接受腎臟紓緩治療，其中位跟進日數為782日。 結果: 大部份病人和家屬之所以拒絕腎透析是由於預計的身體負累，只有百分之一點六接受腎臟紓緩治療會改變主義而接受腎透析。選擇腎透析和腎臟紓緩治療的病人在基本的身體狀況有明顯分別。生存率亦隨著修改版查爾森共病量表的分數而下降。年長、精神自主能力缺欠、高血脂、修改版查爾森共病量表分數高、腎小球濾過率低、白蛋白低均屬重要暨獨立的因素以預計較差的生存率。本文亦會探討在中國文化處境下影響預設照顧計劃討論的因素。 結論: 有組織的預設照顧計劃討論能幫助病人在未期腎病的醫療方向作出知情的選擇

Isogenic Human-Induced Pluripotent Stem-Cell-Derived Cardiomyocytes Reveal Activation of Wnt Signaling Pathways Underlying Intrinsic Cardiac Abnormalities in Rett Syndrome

Rett syndrome (RTT) is a severe neurodevelopmental disorder caused by MeCP2 mutations. Nonetheless, the pathophysiological roles of MeCP2 mutations in the etiology of intrinsic cardiac abnormality and sudden death remain unclear. In this study, we performed a detailed functional studies (calcium and electrophysiological analysis) and RNA-sequencing-based transcriptome analysis of a pair of isogenic RTT female patient-specific induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CMs) that expressed either MeCP2wildtype or MeCP2mutant allele and iPSC-CMs from a non-affected female control. The observations were further confirmed by additional experiments, including Wnt signaling inhibitor treatment, siRNA-based gene silencing, and ion channel blockade. Compared with MeCP2wildtype and control iPSC-CMs, MeCP2mutant iPSC-CMs exhibited prolonged action potential and increased frequency of spontaneous early after polarization. RNA sequencing analysis revealed up-regulation of various Wnt family genes in MeCP2mutant iPSC-CMs. Treatment of MeCP2mutant iPSC-CMs with a Wnt inhibitor XAV939 significantly decreased the β-catenin protein level and CACN1AC expression and ameliorated their abnormal electrophysiological properties. In summary, our data provide novel insight into the contribution of activation of the Wnt/β-catenin signaling cascade to the cardiac abnormalities associated with MeCP2 mutations in RTT