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Background: Overexpression of Aurora-A and mutant Ras (RasV12) together has been detected in human bladder cancer tissue. However, it is not clear whether this phenomenon is a general event or not. Although crosstalk between Aurora-A and Ras signaling pathways has been reported, the role of these two genes acting together in tumorigenesis remains unclear.Methods: Real-time PCR and sequence analysis were utilized to identify Ha- and Ki-ras mutation (Gly -> Val). Immunohistochemistry staining was used to measure the level of Aurora-A expression in bladder and colon cancer specimens. To reveal the effect of overexpression of the above two genes on cellular responses, mouse NIH3T3 fibroblast derived cell lines over-expressing either RasV12 and wild-type Aurora-A (designated WT) or RasV12 and kinase-inactivated Aurora-A (KD) were established. MTT and focus formation assays were conducted to measure proliferation rate and focus formation capability of the cells. Small interfering RNA, pharmacological inhibitors and dominant negative genes were used to dissect the signaling pathways involved.Results: Overexpression of wild-type Aurora-A and mutation of RasV12 were detected in human bladder and colon cancer tissues. Wild-type Aurora-A induces focus formation and aggregation of the RasV12 transformants. Aurora-A activates Ral A and the phosphorylation of AKT as well as enhances the phosphorylation of MEK, ERK of WT cells. Finally, the Ras/MEK/ERK signaling pathway is responsible for Aurora-A induced aggregation of the RasV12 transformants.Conclusion: Wild-type-Aurora-A enhances focus formation and aggregation of the RasV12 transformants and the latter occurs through modulating the Ras/MEK/ERK signaling pathway. [ABSTRACT FROM AUTHOR]

Adult human hearts exhibit limited regenerative capacity. Post-injury cardiomyocyte (CM) loss can lead to myocardial dysfunction and failure. Although neonatal mammalian hearts can regenerate, the underlying molecular mechanisms remain elusive. Herein, comparative transcriptome analyses identify adherens junction protein N-Cadherin as a crucial regulator of CM proliferation/renewal. Its expression correlates positively with mitotic genes and shows an age-dependent reduction. N-Cadherin is upregulated in the neonatal mouse heart following injury, coinciding with increased CM mitotic activities. N-Cadherin knockdown reduces, whereas overexpression increases, the proliferation activity of neonatal mouse CMs and human induced pluripotent stem cell-derived CMs. Mechanistically, N-Cadherin binds and stabilizes pro-mitotic transcription regulator β-Catenin, driving CM self-renewal. Targeted N-Cadherin deletion in CMs impedes cardiac regeneration in neonatal mice, leading to excessive scarring. N-Cadherin overexpression, by contrast, promotes regeneration in adult mouse hearts following ischemic injury. N-Cadherin targeting presents a promising avenue for promoting cardiac regeneration and restoring function in injured adult human hearts., Competing Interests: Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Objective: Physicians often use mobile apps for patient care, but few apps are dedicated to pediatric critical care medicine (PCCM). This study developed a mobile app specifically for Pediatric Critical Care Unit (PICU) residents to aid their onboarding process and aimed to assess whether it could enhance their confidence and comfort levels., Method: From March 2020 to April 2021, 90 residents participated and completed pre- and post-rotation quizzes with 20 questions each. Quiz score differences between the control and app groups were analyzed using t-tests. A survey was also administered at the end of the rotation to compare comfort level and confidence in PCCM knowledge pre- and post-rotation., Results: Enrollment included 50 residents in the control group and 40 in the app group. The participation rate was 100%, but not all participants completed both quizzes and survey. The app group showed a significantly greater improvement in quiz scores from pre- to post-rotation compared to the control group (increase of 0.23 questions vs 1.67, p = 0.045). However, the two groups had no significant differences in confidence in PCCM knowledge (p = 0.246) or comfort levels (p = 0.776) in the PICU., Conclusions: This study found no significant difference in confidence levels between the App and control groups at the end of the PICU rotation. However, the App group outperformed the control group in knowledge assessments. Frequent use of the app likely reinforced essential concepts and facilitated adaptation to the PICU service. Overall, the app's positive impact on knowledge and adaptation indicates it is a valuable tool for enhancing medical residents' educational experiences in busy clinical environments., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2024.)

Adeno-associated virus (AAV) serotypes from primates are being developed and clinically used as vectors for human gene therapy. However, the evolutionary mechanism of AAV variants is far from being understood, except that genetic recombination plays an important role. Furthermore, little is known about the interaction between AAV and its natural hosts, human and nonhuman primates. In this study, natural AAV capsid genes were subjected to systemic evolutionary analysis with a focus on selection drives during the diversification of AAV lineages. A number of positively selected sites were identified from these AAV lineages with functional relevance implied by their localization on the AAV structures. The selection drives of the two AAV2 capsid sites were further investigated in a series of biological experiments. These observations did not support the evolution of the site 410 of the AAV2 capsid driven by selection pressure from the human CD4+ T-cell response. However, positive selection on site 548 of the AAV2 capsid was directly related to host humoral immunity because of the profound effects of mutations at this site on the immune evasion of AAV variants from human neutralizing antibodies at both the individual and population levels. Overall, this work provides a novel interpretation of the genetic diversity and evolution of AAV lineages in their natural hosts, which may contribute to their further engineering and application in human gene therapy., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Background: Recessive GJB2 variants, the most common genetic cause of hearing loss, may contribute to progressive sensorineural hearing loss (SNHL). The aim of this study is to build a realistic predictive model for GJB2-related SNHL using machine learning to enable personalized medical planning for timely intervention., Method: Patients with SNHL with confirmed biallelic GJB2 variants in a nationwide cohort between 2005 and 2022 were included. Different data preprocessing protocols and computational algorithms were combined to construct a prediction model. We randomly divided the dataset into training, validation, and test sets at a ratio of 72:8:20, and repeated this process ten times to obtain an average result. The performance of the models was evaluated using the mean absolute error (MAE), which refers to the discrepancy between the predicted and actual hearing thresholds., Results: We enrolled 449 patients with 2184 audiograms available for deep learning analysis. SNHL progression was identified in all models and was independent of age, sex, and genotype. The average hearing progression rate was 0.61 dB HL per year. The best MAE for linear regression, multilayer perceptron, long short-term memory, and attention model were 4.42, 4.38, 4.34, and 4.76 dB HL, respectively. The long short-term memory model performed best with an average MAE of 4.34 dB HL and acceptable accuracy for up to 4 years., Conclusions: We have developed a prognostic model that uses machine learning to approximate realistic hearing progression in GJB2-related SNHL, allowing for the design of individualized medical plans, such as recommending the optimal follow-up interval for this population., Competing Interests: Declaration of competing interest The below authors declare the following financial interests/personal relationships which may be considered as potential competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Aim: To explore nursing students' perceptions of their design thinking learning experiences on a human development course., Background: Design thinking is a person-centered analytic and creative learning process that promotes higher order thinking skills rather than knowledge retention alone. Currently, this is the first study that has investigated the use of the design thinking process for nursing students on a human development course., Design: A phenomenological research design., Methods: The participants were first-year nursing students enrolled on a human development course at a Taiwanese university. In-depth, semi-structured interviews were conducted in 2022 and sufficiently high information power was obtained after 15 participants were interviewed. Data were systematically analysed, summarized and decoded using Colaizzi's seven analysis steps., Results: Three themes and twelve subthemes emerged from the data. (1) Challenges experienced with the design thinking learning process: participants experienced anxiety because of the unfamiliar assignments, and they found the assignments to be thought-consuming, time-consuming and energy-consuming, plus the students needed to adapt to working in teams with classmates. (2) Adjustment strategies employed by participants: participants highlighted the importance of teamwork during the design thinking learning process. They achieved consensus through communication and sought support from others to overcome challenges related to design thinking; however, they ultimately needed to engage in problem-solving by themselves. (3) Developing the diversity of academic capabilities: after the human development course, participants experienced enriched mindsets, cognitive abilities, problem-solving skills, interpersonal relationships and knowledge application., Conclusions: Design thinking provides creative teaching opportunities and encourages nursing students to engage in experimental and creative learning, which is a meaningful experience for them. Nurse educators could use the insights thus obtained to design a curriculum that sources design thinking as a learning process while facilitating the complexity and diversity of students' higher order thinking skills and not just repetitive learning., Competing Interests: Declaration of Competing Interest None., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study develops a hybrid 3D printing approach that combines fused deposition modeling (FDM) and digital light processing (DLP) techniques for fabricating bioscaffolds, enabling rapid mass production. The FDM technique fabricates outer molds, while DLP prints struts for creating penetrating channels. By combining these components, hydroxyapatite (HA) bioscaffolds with different channel sizes (600, 800, and 1000 μ m) and designed porosities (10%, 12.5%, and 15%) are fabricated using the slurry casting method with centrifugal vacuum defoaming for significant densification. This innovative method produces high-strength bioscaffolds with an overall porosity of 32%-37%, featuring tightly bound HA grains and a layered surface structure, resulting in remarkable cell viability and adhesion, along with minimal degradation rates and superior calcium phosphate deposition. The HA scaffolds show hardness ranging from 1.43 to 1.87 GPa, with increasing compressive strength as the designed porosity and channel size decrease. Compared to human cancellous bone at a similar porosity range of 30%-40%, exhibiting compressive strengths of 13-70 MPa and moduli of 0.8-8 GPa, the HA scaffolds demonstrate robust strengths ranging from 40 to 73 MPa, paired with lower moduli of 0.7-1.23 GPa. These attributes make them well-suited for cancellous bone repair, effectively mitigating issues like stress shielding and bone atrophy., (© 2024 IOP Publishing Ltd.)

This study addresses the pressing need for sustainable bioremediation solutions to combat increasing pollution challenges in alignment with sustainability development goals. The research focuses on developing a co-culture approach involving microalgae and Komagataeibacter europaeus BCRC 14148 bacterium to create a biocomposite for efficient ammonia removal. Nanocellulose, produced by the bacterium, serves as a substrate for microalgae attachment. Optimization using specific growth media ratios resulted in biocomposite yields of 4.05 ± 0.16 g/L and 3.83 ± 0.13 g/L in HS medium with fructose and glucose, respectively. The optimal conditions include a 40:60 ratio of HS-F to TAP medium, 25 ℃ incubation, 6000 Lux light intensity, pH 5.5, and a 48-hour incubation period. When applied to wastewater treatment, the biocomposite demonstrated exceptional ammonium removal efficiency at 91.64 ± 1.27 %. This co-culture-derived biocomposite offers an eco-friendly, recyclable, and effective solution for sustainable environmental bioremediation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background: Depression is a common mental disorder. Literature has explored patients' perspectives of the recovering process of depression. However, there is a lack of research to explore both patients' and caregivers' perspectives of the healing process and develop a theory to support patients with depression., Aims: The purpose of this study is to develop a substantive theory that depicts patients with depression toward healing and recovering., Methods: This study used a grounded theory approach and collected data between 2019 and 2020 in a medical center in Taiwan. A theoretical sampling was performed after interviewing 29 participants, including 20 patients who had recovered from depression and nine caregivers; data saturation was achieved. Data analysis was conducted with open, axial, and selective coding and used NVivo Version 11 to aid the process of coding., Results: A substantive theory was developed and the core category was "Patients' fortitude through the healing and recovering process of depression." Other main categories interrelated in this core category were reframing negative thinking and cultivating positive thinking, rebuilding a positive self-worth by embracing self-compassion, and learning to cope with everyday stress., Conclusions: This theory could help health care professionals to work therapeutically with patients and commend their fortitude while experiencing depression and engaging them with the care they planned together; and find some joy in life. Educators and researchers could use this theory to advance nursing care.

Purpose: Despite the importance of radial endobronchial ultrasound (rEBUS) in transbronchial biopsy, researchers have yet to apply artificial intelligence to the analysis of rEBUS images., Materials and Methods: This study developed a convolutional neural network (CNN) to differentiate between malignant and benign tumours in rEBUS images. This study retrospectively collected rEBUS images from medical centres in Taiwan, including 769 from National Taiwan University Hospital Hsin-Chu Branch, Hsinchu Hospital for model training (615 images) and internal validation (154 images) as well as 300 from National Taiwan University Hospital (NTUH-TPE) and 92 images were obtained from National Taiwan University Hospital Hsin-Chu Branch, Biomedical Park Hospital (NTUH-BIO) for external validation. Further assessments of the model were performed using image augmentation in the training phase and test-time augmentation (TTA)., Results: Using the internal validation dataset, the results were as follows: area under the curve (AUC) (0.88 (95% CI 0.83 to 0.92)), sensitivity (0.80 (95% CI 0.73 to 0.88)), specificity (0.75 (95% CI 0.66 to 0.83)). Using the NTUH-TPE external validation dataset, the results were as follows: AUC (0.76 (95% CI 0.71 to 0.80)), sensitivity (0.58 (95% CI 0.50 to 0.65)), specificity (0.92 (95% CI 0.88 to 0.97)). Using the NTUH-BIO external validation dataset, the results were as follows: AUC (0.72 (95% CI 0.64 to 0.82)), sensitivity (0.71 (95% CI 0.55 to 0.86)), specificity (0.76 (95% CI 0.64 to 0.87)). After fine-tuning, the AUC values for the external validation cohorts were as follows: NTUH-TPE (0.78) and NTUH-BIO (0.82). Our findings also demonstrated the feasibility of the model in differentiating between lung cancer subtypes, as indicated by the following AUC values: adenocarcinoma (0.70; 95% CI 0.64 to 0.76), squamous cell carcinoma (0.64; 95% CI 0.54 to 0.74) and small cell lung cancer (0.52; 95% CI 0.32 to 0.72)., Conclusions: Our results demonstrate the feasibility of the proposed CNN-based algorithm in differentiating between malignant and benign lesions in rEBUS images., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Aneuploidy, the presence of chromosome gains or losses, is a hallmark of cancer. Here, we describe KaryoCreate (karyotype CRISPR-engineered aneuploidy technology), a system that enables the generation of chromosome-specific aneuploidies by co-expression of an sgRNA targeting chromosome-specific CENPA-binding ɑ-satellite repeats together with dCas9 fused to mutant KNL1. We design unique and highly specific sgRNAs for 19 of the 24 chromosomes. Expression of these constructs leads to missegregation and induction of gains or losses of the targeted chromosome in cellular progeny, with an average efficiency of 8% for gains and 12% for losses (up to 20%) validated across 10 chromosomes. Using KaryoCreate in colon epithelial cells, we show that chromosome 18q loss, frequent in gastrointestinal cancers, promotes resistance to TGF-β, likely due to synergistic hemizygous deletion of multiple genes. Altogether, we describe an innovative technology to create and study chromosome missegregation and aneuploidy in the context of cancer and beyond., Competing Interests: Declaration of interests T.D. is on the Scientific Advisory Board (SAB) of io9. S.M.L. is a co-founder of io9 and a member of Biological Dynamics, Inc. SAB. T.D. and N.B. are inventors on patent application no. 63/375,181., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Bacterial cellulose (BC) has attracted worldwide attention owing to its tremendous properties and versatile applications. BC has huge market demand, however; its production is still limited hence important to explore the economically and technically feasible bioprocess for its improved production. The current study is based on improving the bioprocess for BC production employing Komagataeibacter europeaus 14148. Physico-chemical parameters have been optimized e.g., initial pH, incubation temperature, incubation period, inoculum size, and carbon source for maximum BC production. The study employed crude and/or a defined carbon source in the production medium. Hestrin and Schramm (HS) medium was used for BC production with initial pH 5.5 at 30 °C after 7 days of incubation under static conditions. The yield of BC obtained from fruit juice extracted from orange, papaya, mango and banana were higher than other sugars employed. The maximum BC yield of 3.48 ± 0.16 g/L was obtained with papaya extract having 40 g/L reducing sugar concentration and 3.47 ± 0.05 g/L BC was obtained with orange extract having 40 g/L reducing sugar equivalent in the medium. BC yield was about three-fold higher than standard HS medium. Fruit extracts can be employed as sustainable and economic substrates for BC production to replace glucose and fructose., Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05451-y., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© Association of Food Scientists & Technologists (India) 2022.)

Environmental concerns have taken a center stage in our lives driving the society towards biorefinery. Bioprocess development to produce valuable products utilizing waste has its own significance in circular bioeconomy and environmental sustainability. In the present study, production of bacterial cellulose using pineapple waste as carbon source by Komagataeibacter europaeus was undertaken and it was applied for removal of vanadium, a heavy metal which is generated as waste by semiconductors industry in Taiwan. Highest yield of bacterial cellulose (BC) e.i. 5.04 g/L was obtained with pineapple core hydrolysate (HS-PC) replacing glucose in HS medium. The vanadium adsorption capacity by BC produced by HS medium was 5.24 mg/g BC at pH 4 and 2.85 mg/g BC was observed on PCH medium. BC was characterised via SEM, FTIR and XRD., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Aims: Fractures are the result of fragile bone structures after trauma caused by direct or indirect external impact or strong muscular contraction. Most fracture patients undergo surgical fixation to accelerate the healing process and restore the function of mutilated bone. Promoting the healing process remains an important issue for the treatment of bone fractures. Our previous studies demonstrated the remarkable bone-protective effects of kefir peptides (KPs) in ovariectomized rats and mice. In this study, we further evaluate the efficacy of KPs on fracture healing using a rat model of femoral fracture., Main Methods: Fifteen 8-week-old male Sprague Dawley (SD) rats were divided into the sham, mock, and KPs groups, in which the mock and the KPs groups underwent femur-fracture surgery with nail fixation, while the sham group underwent a sham operation. The next day, rats were orally administered with daily 400 mg/kg of KPs (KPs group) or distilled water (sham and mock groups) for four weeks. X-ray imaging, histochemical staining and serum osteogenic markers were applied for fracture healing evaluation. In vitro, mouse bone marrow mesenchymal stem cells (BMMSCs) and MC3T3-E1 line were subjected to osteoblast differentiation in the presence of KPs and compared with no KPs treatment., Key Findings: The results demonstrated that KPs treatment improved the progression of the fracture healing process (p < 0.05) and significantly increased the expressions of Col1a1, Alp, Spp1, Vegfa and Cox2 mRNA in the femurs of the KPs-treated fractured rats compared to those of the mock-treated fracture rats. In vitro, KPs treatment promoted bone regeneration factor (Col1a1, Alp, M-csf and Phospho1) expression in MC3T3-E1-derived osteoblast cultures (on Day 3) and enhanced osteogenic differentiation and mineralization in BMMSC-derived osteoblast cultures (on Day 17 and Day 21)., Significance: This is the first study to show that KPs can help with fracture healing by promoting osteogenic differentiation, and it also suggests that KPs can be used as a nutritional supplement to accelerate fracture healing., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Globally, breast cancer is the most common cancer type in terms of incidence for women. Women with breast cancer endure higher levels of psychological distress than other types of cancer because many lose their identity as a woman, which is an additional characteristic of their psychological distress. Research using phenomenology to explore "the meaning of life" is rare among women with breast cancer., Purpose: The purpose of this study was to explore the perspectives of women with breast cancer on "the meaning of life.", Methods: A phenomenological approach was used. Twenty-six women living with breast cancer drawn from a cancer treatment hospital in Taiwan participated in this study. A semistructured interview was utilized to collect the data, and Colaizzi's seven steps were used to analyze the data., Results: Four themes emerged: (a) value of overcoming suffering, (b) value of reciprocal love, (c) value of self-transcendence, and (d) value of spiritual comfort. This means that the participants defined "the meaning of life" through the lens of suffering from cancer, reciprocal love from their families and friends, uncovering and discovering creative pathways that transformed their pain while searching for the value of their existence, and seeking spiritual guidance from religion., Conclusions: The participants identified the most pivotal aspect of healing as transforming their pain and accepting value for their suffering. They acknowledged they could not change the fact that they had cancer, but they could learn to accept it as part of their lived experience. Healthcare professionals may use these four themes at a clinically appropriate time on women's journeys toward healing to inspire women with breast cancer to process their own unique "meaning of life.", Competing Interests: The authors declare no conflicts of interest., (Copyright © 2022 The Authors. Published by Wolters Kluwer Health, Inc.)

Background: Intensive care management of diabetic ketoacidosis (DKA) is targeted to reverse ketoacidosis, replace the fluid deficit, and correct electrolyte imbalances. Adequate restoration of circulation and treatment of shock is key. Pediatric treatment guidelines of DKA have become standard but complexities arise in children with co-morbidities. Congenital nephrogenic diabetes insipidus (NDI) is a rare hereditary disorder characterized by impaired kidney concentrating ability and treatment is challenging. NDI and DKA together have only been previously reported in one patient., Case Diagnosis/treatment: We present the case of a 12-year-old male with NDI and new onset DKA with hyperosmolality. He presented in hypovolemic shock with altered mental status. Rehydration was challenging and isotonic fluid resuscitation resulted in increased urine output and worsening hyperosmolar state. Use of hypotonic fluid and insulin infusion led to lowering of serum osmolality faster than desired and increased the risk for cerebral edema. Despite the rapid decline in serum osmolality his mental status improved so we allowed him to drink free water mixed with potassium phosphorous every hour to match his urinary output (1:1 replacement) and continued 0.45% sodium chloride based on his fluid deficit and replacement rate with improvement in his clinical status., Conclusions: This case illustrates the challenges in managing hypovolemic shock, hyperosmolality, and extreme electrolyte derangements driven by NDI and DKA, as both disease processes drive excessive urine output, electrolyte and acid-base imbalances, and rapid fluctuation in osmolality., (© 2022. The Author(s), under exclusive licence to International Pediatric Nephrology Association.)

Periodontitis (PD) is a common oral disease associated with various other diseases, particularly those affecting the cardiovascular system. This study explored whether peripheral artery occlusive disease (PAOD) is associated with PD and dental scaling. This study was a retrospective cohort study design from 2000 to 2018. The study population was newly diagnosed with periodontitis. The comparison group was defined as never diagnosed with periodontitis. The outcome variable was defined with the diagnosis of peripheral arterial occlusive disease (PAOD). The propensity score matching was performed by age, sex, comorbidities, and dental scaling between the two groups. Kaplan-Meier analysis was used to calculate the cumulative incidence of PAOD among the two groups. To perform the independent risk of the PAOD group, the multivariate Cox proportional hazard model was used to estimate the hazard ratios. First, 792,681 patients with PD and 458,521 patients with no history of PD were selected from Taiwan's Longitudinal Health Insurance Database, which comprises the data of two million beneficiaries. After propensity score matching between the PD and non-PD groups for age, sex, comorbidities, and dental scaling, 357,106 patients in each group were analyzed for PAOD risk. The incidence density, relative risk, and cumulative incidence of PAOD were higher in the PD group than in the non-PD group. After adjusting for all variables, the risk of PAOD for the PD group was greater than for the non-PD group (adjusted hazard ratio = 1.03; 95% CI, 1.01-1.06). Undergoing at least one dental scaling procedure reduced the risk of PAOD. Age over 65 years was also a risk factor. In conclusion, patients with PD have an increased risk of PAOD. In addition, our results can lead to increased attention to oral hygiene, as dental scaling has a trend towards a lower risk of PAOD.

A bioprospecting study was conducted from Seawater samples collected at Kaohsiung Seacoast, Taiwan. The current research was aimed to isolate potential lutein-producing strain, evaluate and optimize the best cultivation mode, lutein accumulation stage, lutein-extraction method, and condition to recover maximum lutein (main product) and lipid (byproduct). Biorefinery is the latest approach worldwide to extract multi-products for cost-effectiveness. Selected isolate among several isolates, identified as Chlorella sorokiniana Kh12 and exploited under biorefinery concept for lutein and lipid extraction. Kh12 cultivated under mixotrophy: 2X-(HT)-9k yielded maximum biomass (3.46 g L -1 ) and lutein (13.69 mg g -1 ) which is among the higher yields reported so far. Among various tested solvents, methanol was the best extractor. Bead milling was most effective to disrupt algal cell walls, seven minutes of milling was best for maximum lutein (7.56 mg g -1 ) extraction. Kh12 could be a promising candidate for commercial lutein and lipid co-production based on the outcome., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Adeno-associated viruses (AAV) serve as vectors for therapeutic gene delivery. AAV9 vectors have been FDA approved, as Zolgensma, for the treatment of spinal muscular atrophy and are being evaluated in clinical trials for the treatment of neurotropic and musculotropic diseases. A major hurdle for AAV-mediated gene delivery is the presence of preexisting neutralizing antibodies in 40 to 80% of the general population. These preexisting antibodies can reduce therapeutic efficacy through viral neutralization and the size of the patient cohort eligible for treatment. In this study, cryo-electron microscopy and image reconstruction were used to define the epitopes of five anti-AAV9 monoclonal antibodies (MAbs), ADK9, HL2368, HL2370, HL2372, and HL2374, on the capsid surface. Three of these, ADK9, HL2370, and HL2374, bound to or near the icosahedral 3-fold axes, HL2368 bound to the 2/5-fold wall, and HL2372 bound to the region surrounding the 5-fold axes. Pseudoatomic modeling enabled the mapping and identification of antibody contact amino acids on the capsid, including S454 and P659. These epitopes overlap previously defined parvovirus antigenic sites. Capsid amino acids critical for the interactions were confirmed by mutagenesis, followed by biochemical assays testing recombinant AAV9 (rAAV9) variants capable of escaping recognition and neutralization by the parental MAbs. These variants retained parental tropism and had similar or improved transduction efficiency compared to AAV9. These engineered rAAV9 variants could expand the patient cohort eligible for AAV9-mediated gene delivery by avoiding preexisting circulating neutralizing antibodies. IMPORTANCE The use of recombinant adeno-associated viruses (rAAVs) as delivery vectors for therapeutic genes is becoming increasingly popular, especially following the FDA approval of Luxturna and Zolgensma, based on serotypes AAV2 and AAV9, respectively. However, high-titer anti-AAV neutralizing antibodies in the general population exempt patients from treatment. The goal of this study is to circumvent this issue by creating AAV variant vectors not recognized by preexisting neutralizing antibodies. The mapping of the antigenic epitopes of five different monoclonal antibodies (MAbs) on AAV9, to recapitulate a polyclonal response, enabled the rational design of escape variants with minimal disruption to cell tropism and gene expression. This study, which included four newly developed and now commercially available MAbs, provides a platform for the engineering of rAAV9 vectors that can be used to deliver genes to patients with preexisting AAV antibodies.

Melanoma is the cause of most deaths from skin cancer. The extracellular signal-regulated kinase 1/2 (ERK1/2) pathway has been reported to participate in progression of melanoma in fair skinned populations. ERK1/2 is found in both the cytoplasm and nucleus of cells, and phosphorylated ERK1/2 has been implicated in tumor progression. We investigated the relation between melanoma progression and expression of cytoplasmic and nuclear phosphorylated ERK1/2. We examined 34 surgically resected melanomas and investigated their clinicopathologic characteristics. We found immunostaining of phosphorylated ERK1/2 in all melanomas and faint staining in benign nevi. We found expression of cytoplasmic phosphorylated ERK1/2 in most melanomas; however, nuclear phosphorylated ERK1/2 expression was found in only five melanomas. Expression of cytoplasmic phosphorylated ERK1/2 was related to the tumor stage in melanoma. Nine of 10 cases of distant metastasis were positive for cytoplasmic phosphorylated ERK1/2. Our findings suggest that phosphorylated ERK1/2 expression is relevant to clinical pathology and that in melanoma patients, phosphorylated ERK1/2 expression is found in both the cytoplasm and nucleus. Our findings suggest that cytoplasmic phosphorylated ERK1/2 participates in progression of melanoma and that it could be a useful target for clinical treatment of melanoma.

Aim and Objectives: The aim of this study was to develop a theory to help guide patients with end-stage renal disease to adapt to peritoneal dialysis., Background: Taiwan ranks first worldwide in end-stage renal disease incidence and dialysis prevalence. Many patients cannot accept long-term dialysis treatment and thus face several physical and psychological suffering. No theory has yet been developed to help guide patients with end-stage renal disease to adapt to peritoneal dialysis., Design: A grounded theory approach was used in this study., Methods: A theoretical sampling was performed after interviewing 25 patients who had adapted to peritoneal dialysis at a medical centre in Taiwan from January 2018 to September 2018; data saturation was achieved. Data were analysed using open, axial and selective coding and while using the constant comparison technique. COREQ reporting guidelines were utilised., Results: A substantive theory was developed to help guide patients with end-stage renal disease to adapt to peritoneal dialysis. The core category that emerged from the data collected was 'Confronting peritoneal dialysis to live and co-exist with peritoneal dialysis'. Other key categories linked to and embraced in this core category were as follows: positive self-regulation, regulation of daily life and the process of adaptation to dialysis method., Conclusion: The results could help healthcare professionals to better understand the process of end-stage renal disease patients' adaptation to peritoneal dialysis, thereby facilitating patients' adaptation to dialysis in their daily life, enhancing their quality of life and improving the quality of medical care., Relevance to Clinical Practice: Healthcare professionals could use this theory as reference when providing care for peritoneal dialysis patients to assist them in adapting to life with peritoneal dialysis as soon as possible through positive self-regulation, daily life adjustments and the process of adapting to the dialysis method., (© 2021 John Wiley & Sons Ltd.)

Bacterial cellulose (BC) represents a novel bio-origin nonomaterial with its unique properties having diverse applications. Increased market demand and low yield are the major reason for its higher cost. Bacteria belonging to Komagataeibacter sp are the most exploited ones for BC production. Development of a cost-effective bioprocess for higher BC production is desirable. Though static fermentation modes have been majorly employed for BC production using tray fermenters, agitated mode has also been employed successfully with air-lift fermenters as well as stirred tank reactors. Bioprocess advances in recent years has led BC production to an upper level; however, challenges of aeration requirement and labor cost towards the higher end is associated with static cultivation at large scale. We have discussed the bioprocess development for BC production in recent years along with the challenges associated and the path forward., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Although most children with coronavirus disease 2019 (COVID-19) are asymptomatic or only with mild symptoms, many symptomatic children still require admission to the intensive care unit. Multiple cases of diabetic ketoacidosis (DKA) and hyperosmolar hyperglycemic syndrome (HHS) associated with COVID-19 have been reported in adults. However, to our knowledge, only few similar cases have been published in the pediatric population. We report one of the first few severe cases of mixed HHS with DKA associated with COVID-19 in an adolescent. Our patient was successfully treated with intravenous immunoglobulin, Remdesivir, and methylprednisolone. As the pandemic continues, clinicians should be aware of this syndrome and consider early use of Remdesivir and corticosteroids. Further studies are required to understand the pathophysiology of this syndrome occurring with COVID-19., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Yu Shan Tseng et al.)

Freshwater demand is rising worldwide due to largely increasing population and industrialization. Latest focus is to explore the Ocean and saline effluent from industries to produce freshwater in a sustainable way via algal desalination. Current physicochemical desalination technology is not only an energy-intensive and expensive process but also gives severe environmental impact from brine and GHGs emissions. Therefore, it is neither environmentally-friendly nor feasible to countries with limited resources. Biodesalination could be an attractive technology with recent breakthroughs in algal bioprocess with fast growth rate under highly saline conditions to effectively remove salts optimally 50-67% from saline water. Algal desalination mainly occurs through biosorption and bioaccumulation which governs by biotic and abiotic factors e.g., strain, temperature, pH, light and nutrients etc. This review provides a current scenario of this novel technology by an in-depth assessment of technological advancement, social impact, possible risks and scope for policy implications., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This study uses hyperspectral imaging (HSI) and a deep learning diagnosis model that can identify the stage of esophageal cancer and mark the locations. This model simulates the spectrum data from the image using an algorithm developed in this study which is combined with deep learning for the classification and diagnosis of esophageal cancer using a single-shot multibox detector (SSD)-based identification system. Some 155 white-light endoscopic images and 153 narrow-band endoscopic images of esophageal cancer were used to evaluate the prediction model. The algorithm took 19 s to predict the results of 308 test images and the accuracy of the test results of the WLI and NBI esophageal cancer was 88 and 91%, respectively, when using the spectral data. Compared with RGB images, the accuracy of the WLI was 83% and the NBI was 86%. In this study, the accuracy of the WLI and NBI was increased by 5%, confirming that the prediction accuracy of the HSI detection method is significantly improved.

Although rare, children with active coronavirus disease 2019 are at risk of developing malignant arrhythmia. Herein, we present the first paediatric case of refractory ventricular tachycardia from acute fulminant myocarditis secondary to acute COVID-19 infection. This 5-year-old boy required venoarterial extracorporeal membrane oxygenation support, but made a complete recovery without significant morbidity.

Background: The use of virtual simulation methods in nursing education offers potential advantages for students to experience and learn in a safe and realistic environment by acquiring skills and attitudes to prepare for clinical experiences., Objective: The study's aim was to test the hypothesis that nursing students who used a mobile learning app would have significantly (1) higher levels of knowledge about medication administration and nasotracheal suctioning, (2) better development of skill performances on medication administration and nasotracheal suctioning, (3) higher satisfaction, and (4) lower cognitive load than a control group., Design: A randomized experimental study with pretest and posttest design., Settings and Participants: One hundred nursing students were randomly assigned to an experimental or control group at a Southern Taiwan university's school of nursing., Methods: The experimental group received the virtual simulation-based mobile learning app and the control group used traditional paper materials. Participants were assessed on knowledge before and after the intervention. Skill performance and intrinsic and extraneous cognitive load were measured after the intervention., Results: The experimental group who used the mobile app had significantly higher knowledge scores, significantly lower intrinsic and extraneous cognitive load, better skill performance, and higher satisfaction than the control group., Conclusions: The implementation of a mobile app for simulation learning had a positive impact on nursing students' knowledge and skill performances as well as the reduction of learning cognitive loading. Nurse educators and researchers should collaborate in developing virtual learning resources to support clinical nursing education. Tweetable abstract: The mobile app for learning achieved higher knowledge and satisfaction scores, demonstrated better skill performance without increasing cognitive load., Competing Interests: Conflict of Interest The authors declare that they have no competing interests., (Copyright © 2021. Published by Elsevier Ltd.)

Glutamate N -methyl-d-aspartate receptor (NMDAR) hyperfunction is known to contribute to acute renal failure due to ischemia-reperfusion and endotoxemia. d-Serine is a coagonist for NMDAR activation, but whether NMDARs play a role in d-serine-mediated nephrotoxicity remains unclear. Here, we demonstrate that NMDAR blockade ameliorated d-serine-induced renal injury. In NMDAR-expressing LLC-PK 1 cells, which were used as a proximal tubule model, d-serine but not l-serine induced cytotoxicity in a dose-dependent manner, which was abrogated by the selective NMDAR blockers MK-801 and AP-5. Time-dependent oxidative stress, evidenced by gradually increased superoxide and H 2 O 2 production, was associated with d-serine-mediated cytotoxicity; these reactive oxygen species could be alleviated not only after NMDAR inhibition but also by NADPH oxidase (NOX) inhibition. Activation of protein kinase C (PKC)-δ and PKC-ζ is a downstream signal for NMDAR-mediated NOX activation because PKC inhibition diminishes the NOX activity that is induced by d-serine. Renal injury was further confirmed in male Wistar rats that intraperitoneally received d-serine but not l-serine. Peak changes in glucosuria, proteinuria, and urinary excretion of lactate dehydrogenase and malondialdehyde were found after 24 h of treatment. Persistent tubular damage was observed after 7 days of treatment. Cotreatment with the NMDAR blocker MK-801 for 24 h abolished d-serine-induced functional insufficiency and tubular damage. MK-801 attenuated renal superoxide formation by lowering NOX activity and protein upregulation of NOX4 but not NOX2. These results reveal that NMDAR hyperfunction underlies d-serine-induced renal injury via the effects of NOX4 on triggering oxidative stress. NEW & NOTEWORTHY Ionotropic N -methyl-d-aspartate receptors (NMDARs) are not only present in the nervous system but also expressed in the kidney. Overstimulation of renal NMDARs leads to oxidative stress via the signal pathway of calcium/protein kinase C/NADPH oxidase in d-serine-mediated tubular cell damage. Intervention of NMDAR blockade may prevent acute renal injury caused by d-serine.

Background and Objectives: Oral squamous cell carcinoma (OSCC) is a malignant disease with a particularly high incidence in Taiwan. Our objective in this study was to elucidate the involvement of sphingolipid transporter 2 (SPNS2) expression and SPNS2 protein expression in the clinicopathological indexes and the clinical outcomes of OSCC patients. Materials and Methods: Immunohistochemistry analysis was performed for SPNS2 protein expression in samples from 264 cases of OSCC. Correlations of SPNS2 expression with clinicopathological variables and patient survival were analyzed. Results: Our results revealed that the cytoplasmic protein expression of SPNS2 in OSCC tissue specimens was lower than in normal tissue specimens. Negative cytoplasmic protein expression of SPNS2 was significantly correlated with T status and stage. Kaplan-Meier survival curve analysis revealed that negative cytoplasmic SPNS2 expression was predictive of poorer overall survival of OSCC patients in stage III/IV. We also determined that low SPNS2 expression was an independent prognostic factor related to overall survival among OSCC patients in stage III/IV from univariate Cox proportional hazard models. Multivariate Cox proportional hazard models revealed that cytoplasmic SPNS2 expression, T status, lymph node metastasis, and histological grade were independent prognostic factors for survival. Conclusions: Overall, this study determined that SPNS2 protein may be a useful prognostic marker for OSCC patients and potential therapeutic target for OSCC treatment.

Diagnosis of early esophageal neoplasia, including dysplasia and superficial cancer, is a great challenge for endoscopists. Recently, the application of artificial intelligence (AI) using deep learning in the endoscopic field has made significant advancements in diagnosing gastrointestinal cancers. In the present study, we constructed a single-shot multibox detector using a convolutional neural network for diagnosing different histological grades of esophageal neoplasms and evaluated the diagnostic accuracy of this computer-aided system. A total of 936 endoscopic images were used as training images, and these images included 498 white-light imaging (WLI) and 438 narrow-band imaging (NBI) images. The esophageal neoplasms were divided into three classifications: squamous low-grade dysplasia, squamous high-grade dysplasia, and squamous cell carcinoma, based on pathological diagnosis. This AI system analyzed 264 test images in 10 s, and the sensitivity, specificity, and diagnostic accuracy of this system in detecting esophageal neoplasms were 96.2%, 70.4%, and 90.9%, respectively. The accuracy of this AI system in differentiating the histological grade of esophageal neoplasms was 92%. Our system showed better accuracy in diagnosing NBI (95%) than WLI (89%) images. Our results showed the great potential of AI systems in identifying esophageal neoplasms as well as differentiating histological grades.

An artificial intelligence algorithm to detect mycosis fungoides (MF), psoriasis (PSO), and atopic dermatitis (AD) is demonstrated. Results showed that 10 s was consumed by the single shot multibox detector (SSD) model to analyze 292 test images, among which 273 images were correctly detected. Verification of ground truth samples of this research come from pathological tissue slices and OCT analysis. The SSD diagnosis accuracy rate was 93%. The sensitivity values of the SSD model in diagnosing the skin lesions according to the symptoms of PSO, AD, MF, and normal were 96%, 80%, 94%, and 95%, and the corresponding precision were 96%, 86%, 98%, and 90%. The highest sensitivity rate was found in MF probably because of the spread of cancer cells in the skin and relatively large lesions of MF. Many differences were found in the accuracy between AD and the other diseases. The collected AD images were all in the elbow or arm and other joints, the area with AD was small, and the features were not obvious. Hence, the proposed SSD could be used to identify the four diseases by using skin image detection, but the diagnosis of AD was relatively poor.

The parvoviruses are small nonenveloped single stranded DNA viruses that constitute members that range from apathogenic to pathogenic in humans and animals. The infection with a parvovirus results in the generation of antibodies against the viral capsid by the host immune system to eliminate the virus and to prevent re-infection. For members currently either being developed as delivery vectors for gene therapy applications or as oncolytic biologics for tumor therapy, efforts are aimed at combating the detrimental effects of pre-existing or post-treatment antibodies that can eliminate therapeutic benefits. Therefore, understanding antigenic epitopes of parvoviruses can provide crucial information for the development of vaccination applications and engineering novel capsids able to escape antibody recognition. This review aims to capture the information for the binding regions of ∼30 capsid-antibody complex structures of different parvovirus capsids determined to date by cryo-electron microscopy and three-dimensional image reconstruction. The comparison of all complex structures revealed the conservation of antigenic regions among parvoviruses from different genera despite low sequence identity and indicates that the available data can be used across the family for vaccine development and capsid engineering.

Short bowel (SB) increases the risk of kidney stones. However, the underlying mechanism is unclear. Here, we examined how SB affected renal oxalate and citrate handlings for in vivo hyperoxaluric rats and in vitro tubular cells. SB was induced by small intestine resection in male Wistar rats. Sham-operated controls had no resection. After 7 days of recovery, the rats were divided into control, SB (both fed with distilled water), ethylene glycol (EG), and SB+EG (both fed with 0.75% EG for hyperoxaluric induction) groups for 28 days. We collected the plasma, 24 h of urine, kidney, and intestine tissues for analysis. Hypocitraturia was found and persisted up to 28 days for the SB group. Hypocalcemia and high plasma parathyroid hormone (PTH) levels were found in the 28-day SB rats. SB aggravated EG-mediated oxalate nephropathy by fostering hyperoxaluria and hypocitraturia, and increasing the degree of supersaturation and calcium oxalate (CaOx) crystal deposition. These effects were associated with renal up-regulations of the oxalate transporter solute carrier family 26 (Slc26)a6 and citrate transporter sodium-dependent dicarboxylate cotransporter-1 (NaDC-1) but not Slc26a2. The effects of PTH on the SB kidneys were then examined in NRK-52E tubular cells. Recombinant PTH attenuated oxalate-mediated cell injury and up-regulated NaDC-1 via protein kinase A (PKA) activation. PTH, however, showed no additive effects on oxalate-induced Slc26a6 and NaDC-1 up-regulation. Together, these results demonstrated that renal NaDC-1 upregulation-induced hypocitraturia weakened the defense against Slc26a6-mediated hyperoxaluria in SB kidneys for excess CaOx crystal formation. Increased tubular NaDC-1 expression caused by SB relied on PTH., (© 2020 The Author(s).)

Pulmonary fibrosis (PF) is a major public health problem with limited therapeutic options. There is a clear need to identify novel mediators of PF to develop effective therapeutics. Here we show that an ER protein disulfide isomerase, thioredoxin domain containing 5 (TXNDC5), is highly upregulated in the lung tissues from both patients with idiopathic pulmonary fibrosis and a mouse model of bleomycin (BLM)-induced PF. Global deletion of Txndc5 markedly reduces the extent of PF and preserves lung function in mice following BLM treatment. Mechanistic investigations demonstrate that TXNDC5 promotes fibrogenesis by enhancing TGFβ1 signaling through direct binding with and stabilization of TGFBR1 in lung fibroblasts. Moreover, TGFβ1 stimulation is shown to upregulate TXNDC5 via ER stress/ATF6-dependent transcriptional control in lung fibroblasts. Inducing fibroblast-specific deletion of Txndc5 mitigates the progression of BLM-induced PF and lung function deterioration. Targeting TXNDC5, therefore, could be a novel therapeutic approach against PF.

Autophagy is a potential target for the treatment of triple negative breast cancer (TNBC). Because of a lack of targeted therapies for TNBC, it is vital to find optimal agents that avoid chemoresistance and metastasis. Flavopereirine has anti-proliferation ability in cancer cells, but whether it regulates autophagy in breast cancer cells remains unclear. A Premo™ Tandem Autophagy Sensor Kit was used to image the stage at which flavopereirine affects autophagy by confocal microscopy. A plasmid that constitutively expresses p-AKT and siRNA targeting p38 mitogen-activated protein kinase (MAPK) was used to confirm the related signaling pathways by Western blot. We found that flavopereirine induced microtubule-associated protein 1 light chain 3 (LC3)-II accumulation in a dose- and time-dependent manner in MDA-MB-231 cells. Confocal florescent images showed that flavopereirine blocked autophagosome fusion with lysosomes. Western blotting showed that flavopereirine directly suppressed p-AKT levels and mammalian target of rapamycin (mTOR) translation. Recovery of AKT phosphorylation decreased the level of p-p38 MAPK and LC3-II, but not mTOR. Moreover, flavopereirine-induced LC3-II accumulation was partially reduced in MDA-MB-231 cells that were transfected with p38 MAPK siRNA. Overall, flavopereirine blocked autophagy via LC3-II accumulation in autophagosomes, which was mediated by the AKT/p38 MAPK signaling pathway.

Although various advancements in radical surgery and neoadjuvant chemotherapy have been developed in treating osteosarcoma (OS), their clinical prognosis remains poor. A synthetic chemical compound, 3-hydroxylflavone, that is reported to regulate ROS production is known to inhibit human bone osteosarcoma cells. However, its role and mechanism in human OS cells remains unclear. In this study, we have determined the potential of 3-Hydroxy-2-phenylchromone (3-HF) against OS using human osteosarcoma (HOS) cells. Our previous studies showed that Zipper sterile-alpha-motif kinase (ZAK), a kinase member of the MAP3K family, was involved in various cellular events such as cell proliferation and cell apoptosis, and encoded two transcriptional variants, ZAKα and β. In this study, we show that 3-HF induces the expression of ZAK and thereby enhances cellular apoptosis. Using gain of function and loss of function studies, we have demonstrated that ZAK activation by 3-HF in OS cells is confined to a ZAKβ form that presumably plays a leading role in triggering ZAKα expression, resulting in an aggravated cancer apoptosis. Our results also validate ZAKβ as the predominant form of ZAK to drive the anticancer mechanism in HOS cells.

The AAV2.7m8 vector is an engineered capsid with a 10-amino acid insertion in adeno-associated virus (AAV) surface variable region VIII (VR-VIII) resulting in the alteration of an antigenic region of AAV2 and the ability to efficiently transduce retina cells following intravitreal administration. Directed evolution and in vivo screening in the mouse retina isolated this vector. In the present study, we sought to identify the structural differences between a recombinant AAV2.7m8 (rAAV2.7m8) vector packaging a GFP genome and its parental serotype, AAV2, by cryo-electron microscopy (cryo-EM) and image reconstruction. The structures of rAAV2.7m8 and AAV2 were determined to 2.91 and 3.02 Å resolution, respectively. The rAAV2.7m8 amino acid side-chains for residues 219-745 (the last C-terminal residue) were interpretable in the density map with the exception of the 10 inserted amino acids. While observable in a low sigma threshold density, side-chains were only resolved at the base of the insertion, likely due to flexibility at the top of the loop. A comparison to parental AAV2 (ordered from residues 217-735) showed the structures to be similar, except at some side-chains that had different orientations and, in VR-VIII containing the 10 amino acid insertion. VR-VIII is part of an AAV2 antigenic epitope, and the difference is consistent with rAAV2.7m8's escape from a known AAV2 monoclonal antibody, C37-B. The observations provide valuable insight into the configuration of inserted surface peptides on the AAV capsid and structural differences to be leveraged for future AAV vector rational design, especially for retargeted tropism and antibody escape., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: M. Agbandje-McKenna (MAM) is a SAB member for Voyager Therapeutics, Inc., and AGTC, has a sponsored research agreement with AGTC, Intima Bioscience, Inc., and Voyager Therapeutics, and is a consultant for Intima Bioscience, Inc. MAM is a co-founder of StrideBio, Inc. MAM and Antonette Bennett have IP on AAV vectors. A. Keravala (AK), R. Aeran (RA) and YS. Tseng (YST) were employees of Adverum Biotechnology Inc, at the time of this study. Currently, AK is employed by CODA Biotherapeutics, RA by Encoded Therapeutics and YST by Biomarin. M. Gasmi is CSO and president and Brahim Belbellaa employee of Adverum Biotechnologies and both hold stock grants. Adverum Biotechnologies Inc has been licensed the exclusive exploitation rights to the rAAV2.7m8 vector., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Heat stroke (HS) is a life-threatening illness and defined as when body temperature elevates above 40°C accompanied by the systemic inflammatory response syndrome that results in multiple organ dysfunctions. α -Lipoic acid (ALA) acts as a cofactor of mitochondrial enzymes and exerts anti-inflammatory and antioxidant properties in a variety of diseases. This study investigates the beneficial effects of ALA on myocardial injury and organ damage caused by experimental HS and further explores its underlying mechanism. Male Wistar rats were exposed to 42°C until their rectal core temperature reached 42.9°C and ALA was pretreared 40 or 80 mg/kg (i.v.) 1.5 h prior to heat exposure. Results showed that HS-induced lethality and hypothermia were significantly alleviated by ALA treatment that also improved plasma levels of CRE, LDH, and CPK and myocardial injury biomarkers myoglobin and troponin. In addition, ALA reduced cardiac superoxide anion formation and protein expression of cleaved caspase 3 caused by HS. Proinflammatory cytokine TNF- α and NF- κ B pathways were significantly reduced by ALA treatment which may be associated with the upregulation of Hsp70. ALA significantly increased the Atg5-12 complex and LC3B II/LC3B I ratio, whereas the p62 and p-mTOR expression was attenuated in HS rats, indicating the activation of autophagy by ALA. In conclusion, ALA ameliorated the deleterious effects of HS by exerting antioxidative and anti-inflammatory capacities. Induction of Hsp70 and activation of autophagy contribute to the protective effects of ALA in HS-induced myocardial injury., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2019 Hsin-Hsueh Shen et al.)

Breast cancer, which is the most frequently diagnosed cancer, is quite heterogeneous. For breast cancer subtypes lacking targeted therapies, it is vitally essential to find novel agents that prevent chemoresistance and metastatic relapse. Flavopereirine is a β-carboline alkaloid that has antiplasmodial activity, and its antiproliferative effect in different cancers remains unclear. The effect of flavopereirine on cell cycle arrest and apoptosis signaling in breast cancer cells was analyzed by flow cytometry. An inhibitor and siRNA were used to confirm the related signaling pathways by Western blot analysis. We found that flavopereirine caused G0/G1 phase arrest in MCF-7 cells and S phase arrest in MDA-MB-231 cells. MDA-MB-231 cells were more sensitive to flavopereirine-induced apoptosis. Furthermore, we found that flavopereirine-induced apoptosis was partially reduced in MDA-MB-231 cells treated with an extracellular regulated kinase (ERK) inhibitor and p38 mitogen-activated protein kinase (MAPK) siRNA. Moreover, p38 siRNA treatment simultaneously reduced phosphorylated ERK expression levels. Conversely, the recovered phosphorylation of AKT decreased the levels of p-ERK and p-p38 MAPK. Overall, flavopereirine induces cell cycle arrest and the AKT/p38 MAPK/ERK signaling pathway, which contribute to flavopereirine-induced apoptosis in MDA-MB-231 cells., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Osteosarcoma (OS) is a tumor entity that can cause a large number of cancer-related deaths. Although chemotherapy can decrease proliferation and increase apoptosis of human OS cells, the clinical prognosis remains poor. Fisetin is a flavonol found in fruits and vegetables and is reported to inhibit cell growth in numerous cancers. But the molecular mechanism underlying fisetin in human OS cells is not clear. It is known that sterile-alpha motif and leucine zipper containing kinase (ZAK), a kinase in the MAP3K family, is involved in various cell processes, including proliferation and apoptosis. In our lab, we have demonstrated that overexpression of ZAK can induce apoptosis in human OS cells. In the previous studies, MAP4K, the upstream of MAP3K, can act in parallel to MST1/2 to activate LATS1/2 in the Hippo pathway. Turning on the Hippo pathway can decrease proliferation and otherwise cause cell apoptosis in cancer cells. In this study, we found that fisetin can upregulate ZAK expression to induce the Hippo pathway and mediate the activation of JNK/ERK, the downstream of ZAK, to trigger cell apoptosis via AP-1 dependent manner in human OS cells. These findings reveal a novel molecular mechanism underlying fisetin effect on human OS cells., (© 2019 Wiley Periodicals, Inc.)

Climate change impacts on the ecosystem services that people rely on, such as water, air, and agricultural products. The quality and quantity of various ecosystem services may be diminished under conditions of extreme climate change. Therefore, the effects of climate change may be expected to threaten people's health and survival. The concept of health promotion includes attention to the environment and emphasizes balance between nature and manmade structures. In addition, health promotion practices and actions in response to climate change emphasize multidisciplinary cooperation and focus on health inequality and vulnerable populations. Therefore, health promotion professionals must have sufficient professional competence in order to manage the multifaceted health impacts of climate change. The purpose of this article is to review the literature on health promotion and emergency medical care under conditions of climate change. Examples are provided to delineate the biological, psychological, social, and spiritual effects of climate change. The results of this literature review may provide community-based health promotion and emergency medical services guidance for further development and improvements. Healthcare professionals are expected to play a central role in managing the impact of climate change in order to achieve health for all.

ZAK is a novel mixed lineage kinase-like protein that contains a leucine-zipper and a sterile-alpha motif as a protein-protein interaction domain, and it is located in the cytoplasm. There are 2 alternatively spliced forms of ZAK: ZAKα and ZAKβ. Previous studies showed that ZAKα is involved in various cell processes, including cell proliferation, cell differentiation, and cardiac hypertrophy, but the molecular mechanism of ZAKβ is not yet known. In a recent study in our laboratory, we found that ZAKβ can ameliorate the apoptotic effect induced by ZAKα in H9c2 cells. We further hypothesized that ZAKβ could also improve the apoptotic effect induced by ZAKα in human osteosarcoma cells. The results of this study show that ZAKβ can induce apoptosis and decrease cell viability similar to the effects of ZAKα. Interestingly, our ZAKα-specific inhibitor assay shows that the expression of ZAKβ is highly dependent on ZAKα expression. However, ZAKβ expression effectively induces ZAKα expression and results in synergistic enhancement of apoptosis in human osteosarcoma cells. Furthermore, co-immunoprecipitation results revealed that ZAKα can directly interact with ZAKβ, and this interaction may contribute to the enhanced apoptotic effects., Significance of the Study: ZAK is a mixed lineage kinase involved in cell differentiation, proliferation, and hypertrophic growth. ZAKα isoform of ZAK is associated with tumorigenesis, but the function of ZAKβ is not yet known. In H9c2 cells, ZAKβ was found to ameliorate the apoptotic effect induced by ZAKα. However, in osteosarcoma cells, ZAKβ elevates the apoptotic effect induced by ZAKα. In this study, we show that similar to ZAKα, the ZAKβ induces apoptosis and decreases cell viability. Interestingly, the expression of ZAKβ is dependent on ZAKα expression, and ZAKβ further enhances ZAKα expression and results in synergistic enhancement of apoptosis in osteosarcoma cells., (Copyright © 2018 John Wiley & Sons, Ltd.)

Adeno-associated viruses (AAVs) are being developed as vectors for the treatment of genetic disorders. However, pre-existing antibodies present a significant limitation to achieving optimal efficacy for the AAV gene delivery system. Efforts aimed at engineering vectors with the ability to evade the immune response include identification of residues on the virus capsid important for these interactions and changing them. Here K531 is identified as the determinant of monoclonal antibody ADK6 recognition by AAV6, and not the closely related AAV1. The AAV6-ADK6 complex structure was determined by cryo-electron microscopy and the footprint confirmed by cell-based assays. The ADK6 footprint overlaps previously identified AAV antigenic regions and neutralizes by blocking essential cell surface glycan attachment sites. This study thus expands the available repertoire of AAV-antibody information that can guide the design of host immune escaping AAV vectors able to maintain capsid functionality., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Rationale: Cardiac fibrosis plays a critical role in the pathogenesis of heart failure. Excessive accumulation of extracellular matrix (ECM) resulting from cardiac fibrosis impairs cardiac contractile function and increases arrhythmogenicity. Current treatment options for cardiac fibrosis, however, are limited, and there is a clear need to identify novel mediators of cardiac fibrosis to facilitate the development of better therapeutics. Exploiting coexpression gene network analysis on RNA sequencing data from failing human heart, we identified TXNDC5 (thioredoxin domain containing 5), a cardiac fibroblast (CF)-enriched endoplasmic reticulum protein, as a potential novel mediator of cardiac fibrosis, and we completed experiments to test this hypothesis directly., Objective: The objective of this study was to determine the functional role of TXNDC5 in the pathogenesis of cardiac fibrosis., Methods and Results: RNA sequencing and Western blot analyses revealed that TXNDC5 mRNA and protein were highly upregulated in failing human left ventricles and in hypertrophied/failing mouse left ventricle. In addition, cardiac TXNDC5 mRNA expression levels were positively correlated with those of transcripts encoding transforming growth factor β1 and ECM proteins in vivo. TXNDC5 mRNA and protein were increased in human CF (hCF) under transforming growth factor β1 stimulation in vitro. Knockdown of TXNDC5 attenuated transforming growth factor β1-induced hCF activation and ECM protein upregulation independent of SMAD3 (SMAD family member 3), whereas increasing expression of TXNDC5 triggered hCF activation and proliferation and increased ECM protein production. Further experiments showed that TXNDC5, a protein disulfide isomerase, facilitated ECM protein folding and that depletion of TXNDC5 led to ECM protein misfolding and degradation in CF. In addition, TXNDC5 promotes hCF activation and proliferation by enhancing c-Jun N-terminal kinase activity via increased reactive oxygen species, derived from NAD(P)H oxidase 4. Transforming growth factor β1-induced TXNDC5 upregulation in hCF was dependent on endoplasmic reticulum stress and activating transcription factor 6-mediated transcriptional control. Targeted disruption of Txndc5 in mice ( Txndc5 -/- ) revealed protective effects against isoproterenol-induced cardiac hypertrophy, reduced fibrosis (by ≈70%), and markedly improved left ventricle function; post-isoproterenol left ventricular ejection fraction was 59.1±1.5 versus 40.1±2.5 ( P <0.001) in Txndc5 -/- versus wild-type mice, respectively., Conclusions: The endoplasmic reticulum protein TXNDC5 promotes cardiac fibrosis by facilitating ECM protein folding and CF activation via redox-sensitive c-Jun N-terminal kinase signaling. Loss of TXNDC5 protects against β agonist-induced cardiac fibrosis and contractile dysfunction. Targeting TXNDC5, therefore, could be a powerful new therapeutic approach to mitigate excessive cardiac fibrosis, thereby improving cardiac function and outcomes in patients with heart failure., (© 2018 American Heart Association, Inc.)

While Nasopharyngeal carcinoma (NPC) is uncommon in western countries, it is endemic in Southeast Asia and Southern China. Previous study of norcantharidin (NCTD), isolated from blister beetles, has proved its anticancer effect on various tumors. However, the effect of NCTD in NPC has never been studied. The purpose of this study is to inspect the suppression activity of NCTD on NPC, along with the underlying mechanism. NPC cell line NPC-BM was treated with NCTD. NCTD remarkably inhibited proliferation and induce apoptosis in NPC-BM cell. Activation of caspase-3, -8, -9 was observed through western blotting. The expression of antiapoptotic protein Bcl-XL was significantly reduced, but expression of proapoptotic protein Bak was increased after treatment of NCTD. The cytotoxic effect of NCTD on NPC-BM cell is mainly due to apoptosis, mediated by caspase and mitochondrial pathway. These results suggested that NCTD could be a potential anticancer agent for NPC., (© 2017 Wiley Periodicals, Inc.)

Human osteosarcoma (OS) is a malignant cancer of the bone. It exhibits a characteristic malignant osteoblastic transformation and produces a diseased osteoid. A previous study demonstrated that doxorubicin (DOX) chemotherapy decreases human OS cell proliferation and might enhance the relative RNA expression of ZAK. However, the impact of ZAKα overexpression on the OS cell proliferation that is inhibited by DOX and the molecular mechanism underlying this effect are not yet known. ZAK is a protein kinase of the MAPKKK family and functions to promote apoptosis. In our study, we found that ZAKα overexpression induced an apoptotic effect in human OS cells. Treatment of human OS cells with DOX enhanced ZAKα expression and decreased cancer cell viability while increasing apoptosis of human OS cells. In the meantime, suppression of ZAKα expression using shRNA and inhibitor D1771 both suppressed the DOX therapeutic effect. These findings reveal a novel molecular mechanism underlying the DOX effect on human OS cells. Taken together, our findings demonstrate that ZAKα enhances the apoptotic effect and decreases cell viability in DOX-treated human OS cells., (© 2017 Wiley Periodicals, Inc.)

Aims and Objectives: The aim of this study was to explore the factors predicting suicide recovery and to provide guidance for healthcare professionals when caring for individuals who have attempted suicide., Background: The high rate of suicide is a global health problem. Suicide prevention has become an important issue in contemporary mental health. Most suicide research has focused on suicidal prevention and care. There is a lack of research on the factors predicting suicidal recovery., Design: A cross-sectional design was adopted., Methods: A correlational study with a purposive sample of 160 individuals from a suicide prevention centre in southern Taiwan was conducted. The questionnaires included the Brief Symptom Rating Scale-5, Suicidal Recovery Assessment Scale and Beck Hopelessness Scale. Descriptive statistics and linear regressions were used for the analysis., Results: The mean age of the participants was 40.2 years. Many participants were striving to make changes to create a more stable and fulfilling life, had an improved recovery from suicide and had a good ability to adapt or solve problems. The linear regression showed that the Beck Hopelessness Scale scores (ß = -.551, p < .001) and Brief Symptom Rating Scale-5 (ß = -.218, p = .003) and past suicidal behaviour (ß = -.145, p = .008) were significant predictors of individuals' recovery from suicide. They accounted for 57.1% of the variance., Conclusions: Suicidal individuals who have a lower level of hopelessness, a better ability to cope with their mental condition and fewer past suicidal behaviours may better recover from suicide attempts., Relevance to Clinical Practice: The nurses could use the results of this study to predict recovery from suicide in patients with attempted suicide., (© 2017 John Wiley & Sons Ltd.)

Background: Arsenic is a widely distributed metalloid compound that has biphasic effects on cultured cells. In large doses, arsenic can be toxic enough to trigger cell death. In smaller amounts, non-toxic doses may promote cell proliferation and induces carcinogenesis. Aberration of chromosome is frequently detected in epithelial cells and lymphocytes of individuals from arsenic contaminated areas. Overexpression of Aurora-A, a mitotic kinase, results in chromosomal instability and cell transformation. We have reported that low concentration (≦1 μM) of arsenic treatment increases Aurora-A expression in immortalized bladder urothelial E7 cells. However, how arsenic induces carcinogenesis through Aurora-A activation remaining unclear., Methods: Bromodeoxyuridine (BrdU) staining, MTT assay, and flow cytometry assay were conducted to determine cell proliferation. Messenger RNA and protein expression levels of Aurora-A were detected by reverse transcriptional-PCR and Western blotting, respectively. Centrosome of cells was observed by immunofluorescent staining. The transcription factor of Aurora-A was investigated by promoter activity, chromosome immunoprecipitation (ChIP), and small interfering RNA (shRNA) assays. Mouse model was utilized to confirm the relationship between arsenic and Aurora-A., Results: We reveal that low dosage of arsenic treatment increased cell proliferation is associated with accumulated cell population at S phase. We also detected increased Aurora-A expression at mRNA and protein levels in immortalized bladder urothelial E7 cells exposed to low doses of arsenic. Arsenic-treated cells displayed increased multiple centrosome which is resulted from overexpressed Aurora-A. Furthermore, the transcription factor, E2F1, is responsible for Aurora-A overexpression after arsenic treatment. We further disclosed that Aurora-A expression and cell proliferation were increased in bladder and uterus tissues of the BALB/c mice after long-term arsenic (1 mg/L) exposure for 2 months., Conclusion: We reveal that low dose of arsenic induced cell proliferation is through Aurora-A overexpression, which is transcriptionally regulated by E2F1 both in vitro and in vivo. Our findings disclose a new possibility that arsenic at low concentration activates Aurora-A to induce carcinogenesis.