Targeted therapy based on protein–drug conjugates has attracted significant attention owing to its high efficacy and low side effects. However, efficient and stable drug conjugation to a protein ...binder remains a challenge. Herein, a chemoenzymatic method to generate highly stable and homogenous drug conjugates with high efficiency is presented. The approach comprises the insertion of the CaaX sequence at the C‐terminal end of the protein binder, prenylation using farnesyltransferase, and drug conjugation through an oxime ligation reaction. MMAF and an EGFR‐specific repebody are used as the antitumor agent and protein binder, respectively. The method enables the precisely controlled synthesis of repebody–drug conjugates with high yield and homogeneity. The utility of this approach is illustrated by the notable stability of the repebody–drug conjugates in human plasma, negligible off‐target effects, and a remarkable antitumor activity in vivo. The present method can be widely used for generating highly homogeneous and stable PDCs for targeted therapy.
A chemoenzymatic conjugation method that is based on enzymatic prenylation and oxime ligation is a simple and efficient means for generating highly stable and homogeneous protein–drug conjugates in a site‐specific manner. It can be generally applied to the conjugation of drugs to a wide range of protein binders, facilitating the development of targeted therapies with high efficacies and low off‐target effects.
Summary
Compliance with a mandibular advancement device is important for the optimal treatment of obstructive sleep apnea. Recent advances in information and communication technology‐based monitoring ...and intervention for chronic diseases have enabled continuous monitoring and personalized management. Self‐evaluation and self‐regulation through objective monitoring and feedback may improve compliance. The aim of this study was to evaluate the effects of information and communication technology‐based remote monitoring and feedback services, using a smartphone application, on the objective compliance with a mandibular advancement device in patients with obstructive sleep apnea. Forty individuals who were diagnosed with obstructive sleep apnea by polysomnography were randomly assigned to groups A and B. During an initial 6‐week evaluation period, the mandibular advancement device‐wearing time was monitored with the smartphone application in group B, but not in group A. The two groups then switched the monitoring procedures during the second 6‐week period (the smartphone application was then used by group B, but not by group A). If no input data were indicated on the cloud server of the smartphone application during the monitored period, push notifications were provided twice daily. Objective compliance, monitored by a micro‐recorder within the mandibular advancement device, was noted and compared based on whether the monitoring service was provided. The number of mandibular advancement device‐wearing days was significantly higher in the monitored period than in the unmonitored period. The mandibular advancement device‐wearing time did not differ significantly between the two groups. In conclusion, information and communication technology‐based remote monitoring and feedback services demonstrated a potential to increase the objective measures of compliance with mandibular advancement devices.
This study aimed to compare the effectiveness of microcurrent-emitting toothbrushes (MCTs) and ordinary toothbrushes in reducing the dental plaque index (PI) and dental caries activity among ...orthodontic patients. The evaluation was performed using a crossover study design involving 22 orthodontic patients randomly assigned to the MCT or ordinary toothbrush groups. The participants used the designated toothbrush for 4 weeks and had a 1-week wash-out time before crossover to the other toothbrush. PI (Attin's index) and dental caries activity were measured at baseline and at the end of each 4-week period. Additionally, patients completed questionnaires to assess patient satisfaction for "freshness in mouth" and "cleansing degree." The results showed that the MCT group had a significant reduction in PI (p = 0.009), whereas the ordinary toothbrush group did not (p = 0.595). There was no significant difference in the dental caries activity between the two groups (p > 0.05). Patient satisfaction assessment revealed that 65% patients in the MCT group had more than "fair" experience of freshness, in contrast to 50% of patients in the ordinary toothbrush group. Satisfaction with cleansing degree was similar in both groups. Overall, these findings suggest that MCTs are more effective in reducing dental PI than ordinary toothbrushes.
Background
With the rapid increase in the prevalence of cancer worldwide, the utilization of complementary and alternative medicine (CAM) has increased among cancer patients. This review aimed to ...understand the perception, attitudes, and knowledge of healthcare professionals toward using CAM for cancer patients.
Methods
A mixed‐methods systematic review was undertaken in four databases. Inclusion criteria were primary studies reporting perception, attitudes, and knowledge of healthcare professionals for using CAM for cancer patients were eligible. A mixed‐methods convergent synthesis was carried out, and the findings were subjected to a GRADE‐CERQual assessment of confidence.
Results
Forty‐two studies were chosen. The majority of the studies were quantitative and had less than 100 participants. Most publications were from European countries, and oncology was the highest among the specialties. The review found the following themes: feasibility of having negative adverse effects, low expectations of using CAM among HCPs, potential positive effects of using CAM, specific CAM training may be helpful, no concrete regulations to promote CAM practice, and poor physician–patient communication.
Conclusions
Nurses had more positive views than other professions; oncologists were concerned regarding herb–drug interactions; integration of CAM into the healthcare system was favorable; HCPs felt the need to participate in specific CAM training; and HCPs agreed that CAM education should be provided more regularly. Future studies should explore the studies views of cancer patients and details of in‐depth evidence of CAM in oncology settings.
A high diversity of fungi was discovered on various substrates collected at the marine shallow-water Kueishan Island Hydrothermal Vent Field, Taiwan, using culture and metabarcoding methods but ...whether these fungi can grow and play an active role in such an extreme environment is unknown. We investigated the combined effects of different salinity, temperature and pH on growth of ten fungi (in the genera Aspergillus, Penicillium, Fodinomyces, Microascus, Trichoderma, Verticillium) isolated from the sediment and the vent crab Xenograpsus testudinatus. The growth responses of the tested fungi could be referred to three groups: (1) wide pH, salinity and temperature ranges, (2) salinity-dependent and temperature-sensitive, and (3) temperature-tolerant. Aspergillus terreus NTOU4989 was the only fungus which showed growth at 45 °C, pH 3 and 30 ‰ salinity, and might be active near the vents. We also carried out a transcriptome analysis to understand the molecular adaptations of A. terreus NTOU4989 under these extreme conditions. Data revealed that stress-related genes were differentially expressed at high temperature (45 °C); for instance, mannitol biosynthetic genes were up-regulated while glutathione S-transferase and amino acid oxidase genes down-regulated in response to high temperature. On the other hand, hydrogen ion transmembrane transport genes and phenylalanine ammonia lyase were up-regulated while pH-response transcription factor was down-regulated at pH 3, a relative acidic environment. However, genes related to salt tolerance, such as glycerol lipid metabolism and mitogen-activated protein kinase, were up-regulated in both conditions, possibly related to maintaining water homeostasis. The results of this study revealed the genetic evidence of adaptation in A. terreus NTOU4989 to changes of environmental conditions.
Continuous recording of intracellular activities in single cells is required for deciphering rare, dynamic and heterogeneous cell responses, which are missed by population or brief single-cell ...recording. Even if the field of intracellular recording is constantly proceeding, several technical challenges are still remained to conquer this important approach. Here, we demonstrate long-term intracellular recording by combining a vertical nanowire multi electrode array (VNMEA) with optogenetic stimulation to minimally disrupt cell survival and functions during intracellular access and measurement. We synthesized small-diameter and high-aspect-ratio silicon nanowires to spontaneously penetrate into single cells, and used light to modulate the cell's responsiveness. The light-induced intra- and extracellular activities of individual optogenetically-modified cells were measured simultaneously, and each cell showed distinctly different measurement characteristics according to the cell-electrode configuration. Intracellular recordings were achieved continuously and reliably without signal interference and attenuation over 24 hours. The integration of two controllable techniques, vertically grown nanowire electrodes and optogenetics, expands the strategies for discovering the mechanisms for crucial physiological and dynamic processes in various types of cells.
Background and Aim
We conducted a nationwide validation study of diagnostic algorithms to identify cases of inflammatory bowel disease (IBD) within the Korea National Health Insurance System (NHIS) ...database.
Method
Using the NHIS dataset, we developed 44 algorithms combining the International Classification of Diseases (ICD)‐10 codes, codes for Rare and Intractable Diseases (RID) registration and claims data for health care encounters, and pharmaceutical prescriptions for IBD‐specific drugs.
For each algorithm, we compared the case identification results from electronic medical records data with the gold standard (chart‐based diagnosis). A multiple sampling test verified the validation results from the entire study population.
Results
A random nationwide sample of 1697 patients (848 potential cases and 849 negative control cases) from 17 hospitals were included for validation. A combination of the ICD‐10 code, ≥ 1 claims for health care encounters, and ≥ 1 prescription claims (reference algorithm) achieved excellent performance (sensitivity, 93.1% 95% confidence interval 91–94.7; specificity, 98.1% 96.9–98.8; positive predictive value, 97.5% 96.1–98.5; negative predictive value, 94.5% 92.8–95.8) with the lowest error rate (4.2% 3.3–5.3). The multiple sampling test confirmed that the reference algorithm achieves the best performance regarding IBD diagnosis. Algorithms including the RID registration codes exhibited poorer performance compared with that of the reference algorithm, particularly for the diagnosis of patients affiliated with secondary hospitals. The performance of the reference algorithm showed no statistical difference depending on the hospital volume or IBD type, with P‐value < 0.05.
Conclusions
We strongly recommend the reference algorithm as a uniform standard operational definition for future studies using the NHIS database.
Medical disorders caused by second-hand smoke are a major public health concern worldwide. To estimate the level of second-hand smoke exposure, salivary diagnostics for cotinine analysis is a ...compelling alternative in conventional diagnostics using bio-fluids, such as blood and urine, owing to its simple and non-invasive collection method. However, there are several critical issues, such as tedious multisteps, demand for expertise, and field unavailability to collect and transport the purified saliva for further analysis. Here, an all-in-one platform is presented to simply collect real human saliva and directly deliver it onto the biosensing surface. The platform consists of a commercial cotton-swab-type collector, 3D-printed housing, and microfluidic channel integrated with an electrochemical competitive immunosensor to evaluate the level of salivary cotinine. The immunosensor is based on a competitive binding assay between cotinine-conjugated horseradish peroxidase (C-HRP) and cotinine for anti-cotinine binding sites. The current responses obtained from the HRP-thionine-H
O
system decreased proportionally to the cotinine concentration. This immunosensor successfully detected its target over a range of 1 × 10
to 1 × 10
pg ml
with a low limit of detection of 6 × 10
pg ml
and a limit of quantification of 1 × 10
pg ml
. In addition, the platform is applicable to various commercial cotton-swab-type saliva collectors and can successfully transfer the saliva in wide flow rates ranging from 0.1 to 30 ml min
without leakage or damage to the sensing surface. Furthermore, the practicality of the proposed platform was evaluated by measuring cotinine in real human saliva from eight non-smokers. The concentration of cotinine was from 45.7 to 890.8 pg ml
, which was in good agreement with that measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The introduced all-in-one platform represented a reliable performance delivering simple and practical steps in salivary diagnostics.
•RNA adjuvant was developed from the CrPV intergenic region IRES.•The RNA adjuvant functioned as an adjuvant with protein-based vaccines.•The RNA adjuvant increased vaccine efficacy and induced ...balanced Th1/Th2 response.•The RNA adjuvant enhanced APC chemotaxis.
An ideal adjuvant should increase vaccine efficacy through balanced Th1/Th2 responses and be safe to use. Recombinant protein-based vaccines are usually formulated with aluminum (alum)-based adjuvants to ensure an adequate immune response. However, use of alum triggers a Th2-biased immune induction, and hence is not optimal. Although the adjuvanticity of RNA has been reported, a systematic and overall investigation on its efficacy is lacking. We found that single strand RNA (termed RNA adjuvant) derived from cricket paralysis virus intergenic region internal ribosome entry site induced the expression of various adjuvant-function-related genes, such as type 1 and 2 interferon (IFN) and toll-like receptor (TLR), T cell activation, and leukocyte chemotaxis in human peripheral blood mononuclear cells; furthermore, its innate and IFN transcriptome profile patterns were similar to those of a live-attenuated yellow fever vaccine. This suggests that protein-based vaccines formulated using RNA adjuvant function as live-attenuated vaccines. Application of the RNA adjuvant in mouse enhanced the efficacy of Middle East respiratory syndrome spike protein, a protein-subunit vaccine and human papillomavirus L1 protein, a virus-like particle vaccine, by activating innate immune response through TLR7 and enhancing pAPC chemotaxis, leading to a balanced Th1/Th2 responses. Moreover, the combination of alum and the RNA adjuvant synergistically induced humoral and cellular immune responses and endowed long-term immunity. Therefore, RNA adjuvants have broad applicability and can be used with all conventional vaccines to improve vaccine efficacy qualitatively and quantitively.
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