Gene Therapy Using Efficient Direct Lineage Reprogramming Technology for Neurological Diseases.

Gene therapy is an innovative approach in the field of regenerative medicine. This therapy entails the transfer of genetic material into a patient's cells to treat diseases. In particular, gene therapy for neurological diseases has recently achieved significant progress, with numerous studies investigating the use of adeno-associated viruses for the targeted delivery of therapeutic genetic fragments. This approach has potential applications for treating incurable diseases, including paralysis and motor impairment caused by spinal cord injury and Parkinson's disease, and it is characterized by dopaminergic neuron degeneration. Recently, several studies have explored the potential of direct lineage reprogramming (DLR) for treating incurable diseases, and highlighted the advantages of DLR over conventional stem cell therapy. However, application of DLR technology in clinical practice is hindered by its low efficiency compared with cell therapy using stem cell differentiation. To overcome this limitation, researchers have explored various strategies such as the efficiency of DLR. In this study, we focused on innovative strategies, including the use of a nanoporous particle-based gene delivery system to improve the reprogramming efficiency of DLR-induced neurons. We believe that discussing these approaches can facilitate the development of more effective gene therapies for neurological disorders.

Clinical Trends in Management of Locally Advanced ESCC: Real-World Evidence from a Large Single-Center Cohort Study.

Neoadjuvant chemoradiation followed by surgery (NCRT+S) has been widely applied to patients with locally advanced esophageal squamous cell carcinoma (ESCC); however, treatment trends and their survival outcomes in a real-world clinical setting are poorly understood. This study aimed to analyze real-world evidence to understand treatment patterns and outcomes for patients with ESCC. We analyzed the treatment pattern and 5-year overall survival (5yOS) by synthesizing the individuals' general characteristics, cancer information, and treatment records extracted from the Clinical Data Warehouse from 1994 to 2018. Of a total of 2151 patients, most patients received upfront surgery and 5yOS was 36.8% (31.4−43.1%). From 2003 to 2012, the use of NCRT increased, and 5yOS was improved to 42.2% (38.8−45.7%). Notably, after 2013, the proportion of NCRT+S markedly increased up to >50% of patients: 5yOS was much improved to 56.3% (53.2−59.6%). With regard to treatment, patients with NCRT+S had the most favorable 5yOS of 58.1% (53−63.7%), although that for patients with upfront surgery was 48.6% (45.9−51.5%, p < 0.001). Moreover, patients who received adjuvant therapy after surgery had better OS than those with surgery alone (58.4% (52.7−64.7%) vs. 47.3% (44.1−50.7%), p < 0.001). This analysis of real-world data demonstrated a significantly improved survival outcome for locally advanced ESCC over time since NCRT prior to surgery had been routinely applied. We revealed that NCRT+S was the most effective treatment for locally advanced ESCC and that adjuvant chemotherapy may be an encouraging therapeutic option for patients with positive nodes after upfront surgery.

Identifying the Risk of Sepsis in Patients With Cancer Using Digital Health Care Records: Machine Learning-Based Approach.

BACKGROUND: Sepsis is diagnosed in millions of people every year, resulting in a high mortality rate. Although patients with sepsis present multimorbid conditions, including cancer, sepsis predictions have mainly focused on patients with severe injuries. OBJECTIVE: In this paper, we present a machine learning-based approach to identify the risk of sepsis in patients with cancer using electronic health records (EHRs). METHODS: We utilized deidentified anonymized EHRs of 8580 patients with cancer from the Samsung Medical Center in Korea in a longitudinal manner between 2014 and 2019. To build a prediction model based on physical status that would differ between sepsis and nonsepsis patients, we analyzed 2462 laboratory test results and 2266 medication prescriptions using graph network and statistical analyses. The medication relationships and lab test results from each analysis were used as additional learning features to train our predictive model. RESULTS: Patients with sepsis showed differential medication trajectories and physical status. For example, in the network-based analysis, narcotic analgesics were prescribed more often in the sepsis group, along with other drugs. Likewise, 35 types of lab tests, including albumin, globulin, and prothrombin time, showed significantly different distributions between sepsis and nonsepsis patients (P<.001). Our model outperformed the model trained using only common EHRs, showing an improved accuracy, area under the receiver operating characteristic (AUROC), and F1 score by 11.9%, 11.3%, and 13.6%, respectively. For the random forest-based model, the accuracy, AUROC, and F1 score were 0.692, 0.753, and 0.602, respectively. CONCLUSIONS: We showed that lab tests and medication relationships can be used as efficient features for predicting sepsis in patients with cancer. Consequently, identifying the risk of sepsis in patients with cancer using EHRs and machine learning is feasible.

Correction: Candidemia Risk Prediction (CanDETEC) Model for Patients With Malignancy: Model Development and Validation in a Single-Center Retrospective Study.

[This corrects the article DOI: 10.2196/24651.].

Candidemia Risk Prediction (CanDETEC) Model for Patients With Malignancy: Model Development and Validation in a Single-Center Retrospective Study.

BACKGROUND: Appropriate empirical treatment for candidemia is associated with reduced mortality; however, the timely diagnosis of candidemia in patients with sepsis remains poor. OBJECTIVE: We aimed to use machine learning algorithms to develop and validate a candidemia prediction model for patients with cancer. METHODS: We conducted a single-center retrospective study using the cancer registry of a tertiary academic hospital. Adult patients diagnosed with malignancies between January 2010 and December 2018 were included. Our study outcome was the prediction of candidemia events. A stratified undersampling method was used to extract control data for algorithm learning. Multiple models were developed-a combination of 4 variable groups and 5 algorithms (auto-machine learning, deep neural network, gradient boosting, logistic regression, and random forest). The model with the largest area under the receiver operating characteristic curve (AUROC) was selected as the Candida detection (CanDETEC) model after comparing its performance indexes with those of the Candida Score Model. RESULTS: From a total of 273,380 blood cultures from 186,404 registered patients with cancer, we extracted 501 records of candidemia events and 2000 records as control data. Performance among the different models varied (AUROC 0.771- 0.889), with all models demonstrating superior performance to that of the Candida Score (AUROC 0.677). The random forest model performed the best (AUROC 0.889, 95% CI 0.888-0.889); therefore, it was selected as the CanDETEC model. CONCLUSIONS: The CanDETEC model predicted candidemia in patients with cancer with high discriminative power. This algorithm could be used for the timely diagnosis and appropriate empirical treatment of candidemia.

Dysfunction of X-linked inhibitor of apoptosis protein (XIAP) triggers neuropathological processes via altered p53 activity in Huntington's disease.

Mitochondrial dysfunction is associated with neuronal damage in Huntington's disease (HD), but the precise mechanism of mitochondria-dependent pathogenesis is not understood yet. Herein, we found that colocalization of XIAP and p53 was prominent in the cytosolic compartments of normal subjects but reduced in HD patients and HD transgenic animal models. Overexpression of mutant Huntingtin (mHTT) reduced XIAP levels and elevated mitochondrial localization of p53 in striatal cells in vitro and in vivo. Interestingly, XIAP interacted directly with the C-terminal domain of p53 and decreased its stability via autophagy. Overexpression of XIAP prevented mitochondrially targeted-p53 (Mito-p53)-induced mitochondrial oxidative stress and striatal cell death, whereas, knockdown of XIAP exacerbated Mito-p53-induced neuronal damage in vitro. In vivo transduction of AAV-shRNA XIAP in the dorsal striatum induced rapid onset of disease and reduced the lifespan of HD transgenic (N171-82Q) mice compared to WT littermate mice. XIAP dysfunction led to ultrastructural changes of the mitochondrial cristae and nucleus morphology in striatal cells. Knockdown of XIAP exacerbated neuropathology and motor dysfunctions in N171-82Q mice. In contrast, XIAP overexpression improved neuropathology and motor behaviors in both AAV-mHTT-transduced mice and N171-82Q mice. Our data provides a molecular and pathological mechanism that deregulation of XIAP triggers mitochondria dysfunction and other neuropathological processes via the neurotoxic effect of p53 in HD. Together, the XIAP-p53 pathway is a novel pathological marker and can be a therapeutic target for improving the symptoms in HD.

Fracture resistance of teeth obturated with two different types of mineral trioxide aggregate cements / Resistência à fratura de dentes obturados com dois tipos diferentes de cimentos agregados de trióxido minera

Objective: Endodontically obturated teeth have lower fracture resistance depending on the obturating material and technique. The purpose of this study was therefore to evaluate the influence of ProRoot MTA (Dentsply Sirona, Tulsa Division) and OrthoMTA III (BioMTA, Daejeon, Korea) as an obturating material on the fracture resistance of endodontically treated teeth. Material and Methods: Thirty extracted human maxillary central incisors were decoronated and instrumented using Protaper instruments (size F5). Irrigation was performed with 2.5% sodium hypochlorite between each instrument change followed by 7% maleic acid for one minute. Finally, canals were flushed with 5 ml of PBS solution for one minute. Samples were then divided into three groups. Group I- positive control (no root canal filling); Group II- obturation with ProRoot MTA; Group III- obturation with OrthoMTA III. Ten teeth were randomly selected as a negative control in which no treatment was performed. All the specimens were then subjected to fracture strength testing using universal testing machine. For evaluation of biomineralization, six maxillary central incisors were divided into two groups. Group I obturated with ProRoot MTA and group II obturated with OrthoMTA III. These samples were subjected to SEM analysis. Results: Positive control group demonstrated the least fracture resistance, while OrthoMTA III group showed the highest fracture resistance. There was no significant difference between negative control group and ProRoot MTA groups (p=0.821). OrthoMTA III group showed better tubular biomineralization when compared to ProRoot MTA. Conclusions: Root canals obturated with OrthoMTA III had better fracture resistance and increased tubular biomineralization compared to ProRoot MTA. Since root canals obturated with OrthoMTA III had better fracture resistance, it can be used as a promising obturating material.(AU)

Objetivo: Dentes obturados endodonticamente apresentam menor resistência à fratura, dependendo do material e da técnica de obturação. Portanto, o objetivo deste estudo foi avaliar a influência do ProRoot MTA (Dentsply Sirona, Tulsa Division) e OrthoMTA III (BioMTA, Daejeon, Coréia) como material obturador na resistência à fratura de dentes tratados endodonticamente. Material e Métodos: Trinta incisivos centrais superiores humanos extraídos foram decoronados e instrumentados com instrumentos Protaper (tamanho F5). A irrigação foi realizada com hipoclorito de sódio a 2,5% entre cada troca de instrumento, seguida por ácido maleico a 7% por um minuto. Finalmente, os canais foram lavados com 5 ml de solução de PBS por um minuto. As amostras foram então divididas em três grupos. Grupo I - controle positivo(sem preenchimento do canal radicular); Grupo II - obturação com ProRoot MTA; Grupo III -obturação com OrthoMTA III. Dez dentes foram selecionados aleatoriamente como controle negativo, no qual nenhum tratamento foi realizado. Todas as amostras foram então submetidas atestes de resistência à fratura usando uma máquina de teste universal. Para avaliação da biomineralização, seis incisivos centrais superiores foram divididos em dois grupos: grupo Iobturado com ProRoot MTA e grupo II obturado com OrthoMTA III. Essas amostras foram submetidas à análise SEM. Resultados: O grupo controle positivo demonstrou a menor resistência à fratura, enquanto o grupo OrthoMTA III apresentou a maior resistência à fratura. Não houve diferença significativa entre o grupo controle negativo e os grupos ProRoot MTA (p= 0,821). O grupo OrthoMTA III apresentou melhor biomineralização tubular quando comparado ao ProRoot MTA. Conclusões: Os canais radiculares obturados com OrthoMTA III apresentaram melhor resistência à fratura e maior biomineralização tubular em comparação como ProRoot MTA. Como os canais radiculares obturados com OrthoMTA III apresentaram melhor resistência à fratura, podendo ser utilizado como um material obturador promissor.(AU)

Experience of Emergency Department Patients With Using the Talking Pole Device: Prospective Interventional Descriptive Study.

BACKGROUND: Patient engagement is important. However, it can be difficult in emergency departments (EDs). OBJECTIVE: The aim of this study was to evaluate the satisfaction of ED patients using a patient-friendly health information technology (HIT) device, the "Talking Pole," and to assess the factors relevant to their satisfaction. METHODS: This study was conducted in May 2017 at the ED of a tertiary hospital. The "Talking Pole" is a smartphone-based device attached to a intravenous infusion pole with sensors. It is capable of sensing patient movement and fluid dynamics. In addition, it provides clinical information from electronic medical records to patients and serves as a wireless communication tool between patients and nurses. Patients and caregivers who entered the observation room of the ED were selected for the study. The "Talking Pole" devices were provided to all participants, regardless of their need for an intravenous pole upon admittance to the ED. After 2 hours, each participant was given an 18-item questionnaire created for this research, measured on a 5-point Likert scale, regarding their satisfaction with "Talking Pole." RESULTS: Among 52 participants recruited, 54% (28/52) were patients and the remaining were caregivers. In total, 38% (20/52) were male participants; the average age was 54.6 (SD 12.9) years, and 63% (33/52) of the participants were oncology patients and their caregivers. The overall satisfaction rate was 4.17 (SD 0.79 ) points. Spearman correlation coefficient showed a strong association of "overall satisfaction" with "comparison to the previous visit" (ρ=.73 ), "perceived benefit" (ρ=.73), "information satisfaction" (ρ=.70), and "efficiency" (ρ=.70). CONCLUSIONS: In this study, we introduced a patient-friendly HIT device, the "Talking Pole." Its architecture focused on enhancing information delivery, which is regarded as a bottleneck toward achieving patient engagement in EDs. Patient and caregiver satisfaction with the "Talking Pole" was positive in the ED environment. In particular, correlation coefficient results improved our understanding about patients' satisfaction, HIT devices, and services used in the ED.

Homogeneous generation of iDA neurons with high similarity to bona fide DA neurons using a drug inducible system.

Recent work generating induced dopaminergic (iDA) neurons using direct lineage reprogramming potentially provides a novel platform for the study and treatment Parkinson's disease (PD). However, one of the most important issues for iDA-based applications is the degree to which iDA neurons resemble the molecular and functional properties of their endogenous DA neuron counterparts. Here we report that the homogeneity of the reprogramming gene expression system is critical for the generation of iDA neuron cultures that are highly similar to endogenous DA neurons. We employed an inducible system that carries iDA-inducing factors as defined transgenes for direct lineage reprogramming to iDA neurons. This system circumvents the need for viral transduction, enabling a more efficient and reproducible reprogramming process for the generation of genetically homogenous iDA neurons. We showed that this inducible system generates iDA neurons with high similarity to their bona fide in vivo counterparts in comparison to direct infection methods. Thus, our results suggest that homogenous expression of exogenous genes in direct lineage reprogramming is critical for the generation of high quality iDA neuron cultures, making such culture systems a valuable resource for iDA-based drug screening and, ultimately, potential therapeutic intervention in PD.

Nanogrooved substrate promotes direct lineage reprogramming of fibroblasts to functional induced dopaminergic neurons.

The generation of dopaminergic (DA) neurons via direct lineage reprogramming can potentially provide a novel therapeutic platform for the study and treatment of Parkinson's disease. Here, we showed that nanoscale biophysical stimulation can promote the direct lineage reprogramming of somatic fibroblasts to induced DA (iDA) neurons. Fibroblasts that were cultured on flat, microgrooved, and nanogrooved substrates responded differently to the patterned substrates in terms of cell alignment. Subsequently, the DA marker expressions, acquisition of mature DA neuronal phenotypes, and the conversion efficiency were enhanced mostly on the nanogrooved substrate. These results may be attributed to specific histone modifications and transcriptional changes associated with mesenchymal-to-epithelial transition. Taken together, these results suggest that the nanopatterned substrate can serve as an efficient stimulant for direct lineage reprogramming to iDA neurons, and its effectiveness confirms that substrate nanotopography plays a critical role in the cell fate changes during direct lineage reprogramming.