Frequent nonhomologous replacement of replicative helicase loaders by viruses in Vibrionaceae.

Several microbial genomes lack textbook-defined essential genes. If an essential gene is absent from a genome, then an evolutionarily independent gene of unknown function complements its function. Here, we identified frequent nonhomologous replacement of an essential component of DNA replication initiation, a replicative helicase loader gene, in Vibrionaceae. Our analysis of Vibrionaceae genomes revealed two genes with unknown function, named vdhL1 and vdhL2, that were substantially enriched in genomes without the known helicase-loader genes. These genes showed no sequence similarities to genes with known function but encoded proteins structurally similar with a viral helicase loader. Analyses of genomic syntenies and coevolution with helicase genes suggested that vdhL1/2 encodes a helicase loader. The in vitro assay showed that Vibrio harveyi VdhL1 and Vibrio ezurae VdhL2 promote the helicase activity of DnaB. Furthermore, molecular phylogenetics suggested that vdhL1/2 were derived from phages and replaced an intrinsic helicase loader gene of Vibrionaceae over 20 times. This high replacement frequency implies the host's advantage in acquiring a viral helicase loader gene.

Screening for Major Depressive Disorder Using a Wearable Ultra-Short-Term HRV Monitor and Signal Quality Indices.

To encourage potential major depressive disorder (MDD) patients to attend diagnostic sessions, we developed a novel MDD screening system based on sleep-induced autonomic nervous responses. The proposed method only requires a wristwatch device to be worn for 24 h. We evaluated heart rate variability (HRV) via wrist photoplethysmography (PPG). However, previous studies have indicated that HRV measurements obtained using wearable devices are susceptible to motion artifacts. We propose a novel method to improve screening accuracy by removing unreliable HRV data (identified on the basis of signal quality indices (SQIs) obtained by PPG sensors). The proposed algorithm enables real-time calculation of signal quality indices in the frequency domain (SQI-FD). A clinical study conducted at Maynds Tower Mental Clinic enrolled 40 MDD patients (mean age, 37.5 ± 8.8 years) diagnosed on the basis of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition, and 29 healthy volunteers (mean age, 31.9 ± 13.0 years). Acceleration data were used to identify sleep states, and a linear classification model was trained and tested using HRV and pulse rate data. Ten-fold cross-validation showed a sensitivity of 87.3% (80.3% without SQI-FD data) and specificity of 84.0% (73.3% without SQI-FD data). Thus, SQI-FD drastically improved sensitivity and specificity.

Effects of Zinc Acetate Hydrate Supplementation on Renal Anemia with Hypozincemia in Hemodialysis Patients.

INTRODUCTION AND AIMS: This study examined whether zinc supplementation with zinc acetate hydrate improved renal anemia with hypozincemia in patients undergoing hemodialysis. METHODS: The study participants included 21 patients undergoing hemodialysis who presented with a serum zinc level < 60 mg/dL and who were administered zinc acetate hydrate at 50 mg (reduced to 25 mg, as appropriate) for 6 months. Patients with a hemorrhagic lesion, acute-phase disease (pneumonia or cardiac failure), or hematologic disease and those whose treatment was switched from peritoneal dialysis to hemodialysis were excluded. The changes in the erythropoietin resistance index (ERI) before and after zinc acetate hydrate administration were examined. ERI was defined as the dose (IU) of erythropoiesis-stimulating agent (ESA)/week/body weight (kg)/hemoglobin content (g/dL). The differences between the two groups were analyzed using the Wilcoxon signed rank sum test, and p < 0.05 was considered statistically significant. RESULTS: The study participants included 19 men and 2 women aged 41-95 years (mean ± standard deviation (SD): 67.1 ± 13.6). The changes in the values of parameters measured before and after zinc acetate hydrate administration were as follows: Blood Hb did not change significantly, from 10.0-13.6 g/dL (11.5 ± 1.0 g/dL) to 10.2-12.4 g/dL (11.4 ± 0.7 g/dL); serum zinc concentration significantly increased, from 33.0-59.0 mg/dL µg/dL (52.4 ± 7.6 mg/dL µg/dL) to 57.0-124.0 mg/dL µg/dL (84.1 ± 16.3 mg/dL µg/dL; p < 0.01); the ESA dose significantly decreased, from 0-12,000 IU/week (5630 ± 3351 IU/week) to 0-9000 IU/week (4428 ± 2779; p = 0.04); and ERI significantly decreased, from 0.0-18.2 (8.1 ± 5.1) to 0.0-16.0 (6.3 ± 4.3; p = 0.04). CONCLUSIONS: Zinc supplementation increased the serum zinc concentration and significantly reduced the ESA dose and ERI, suggesting that a correction of hypozincemia contributes to lessening renal anemia in these patients.

Development of a highly sensitive method for the quantitative analysis of modified nucleosides using UHPLC-UniSpray-MS/MS.

There are more than 150 types of naturally occurring modified nucleosides, which are believed to be involved in various biological processes. Recently, an ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) technique has been developed to measure low levels of modified nucleosides. A comprehensive analysis of modified nucleosides will lead to a better understanding of intracellular ribonucleic acid modification, but this analysis requires high-sensitivity measurements. In this perspective, we established a highly sensitive and quantitative method using the newly developed ion source, UniSpray. A mass spectrometer was used with a UniSpray source in positive ion mode. Our UHPLC-UniSpray-MS/MS methodology separated and detected the four major nucleosides, 42 modified nucleosides, and dG15N5 (internal standard) in 15 min. The UniSpray method provided good correlation coefficients (>0.99) for all analyzed nucleosides, and a wide range of linearity for 35 of the 46 nucleosides. Additionally, the accuracy and precision values satisfied the criteria of <15% for higher concentrations and <20% for the lowest concentrations of all nucleosides. We also investigated whether this method could measure nucleosides in biological samples using mouse tissues and non-small cell lung cancer clinical specimens. We were able to detect 43 and 31 different modified nucleosides from mouse and clinical tissues, respectively. We also found significant differences in the levels of N6-methyl-N6-threonylcarbamoyladenosine (m6t6A), 1-methylinosine (m1I), 2'-O-methylcytidine (Cm), 5-carbamoylmethyluridine (ncm5U), 5-methoxycarbonylmethyl-2-thiouridine (mcm5S2U), and 5-methoxycarbonylmethyl-2'-O-methyluridine (mcm5Um) between cancerous and noncancerous tissues. In conclusion, we developed a highly sensitive methodology using UHPLC-UniSpray-MS/MS to simultaneously detect and quantify modified nucleosides, which can be used for analysis of biological samples.

Development and Clinical Application of a Novel Non-contact Early Airflow Limitation Screening System Using an Infrared Time-of-Flight Depth Image Sensor.

Obstructive pulmonary diseases, such as diffuse panbronchiolitis (DPB), asthma, chronic obstructive pulmonary disease (COPD), and asthma COPD overlap syndrome (ACOS) trigger a severe reaction at some situations. Detecting early airflow limitation caused by diseases above is critical to stop the progression. Thus, there is a need for tools to enable self-screening of early airflow limitation at home. Here, we developed a novel non-contact early airflow limitation screening system (EAFL-SS) that does not require calibration to the individual by a spirometer. The system is based on an infrared time-of-flight (ToF) depth image sensor, which is integrated into several smartphones for photography focusing or augmented reality. The EAFL-SS comprised an 850 nm infrared ToF depth image sensor (224 × 171 pixels) and custom-built data processing algorithms to visualize anterior-thorax three-dimensional motions in real-time. Multiple linear regression analysis was used to determine the amount of air compulsorily exhaled after maximal inspiration (referred to as the forced vital capacity, FVC EAFL -SS) from the ToF-derived anterior-thorax forced vital capacity (FVC), height, and body mass index as explanatory variables and spirometer-derived FVC as the objective variable. The non-contact measurement is automatically started when an examinee is sitting 35 cm away from the EAFL-SS. A clinical test was conducted with 32 COPD patients (27/5 M/F, 67-93 years) as typical airflow limitation cases recruited at St. Marianna University Hospital and 21 healthy volunteers (10/11 M/F, 23-79 years). The EAFL-SS was used to monitor the respiration of examinees during forced exhalation while sitting still, and a spirometer was used simultaneously as a reference. The forced expiratory volume in 1 s (FEV1% EAFL -SS) was evaluated as a percentage of the FVC EAFL -SS, where values less than 70% indicated suspected airflow limitation. Leave-one-out cross-validation analysis revealed that this system provided 81% sensitivity and 90% specificity. Further, the FEV1 EAFL -SS values were closely correlated with that measured using a spirometer (r = 0.85, p < 0.0001). Hence, EAFL-SS appears promising for early airflow limitation screening at home.

Infection Screening System Using Thermography and CCD Camera with Good Stability and Swiftness for Non-contact Vital-Signs Measurement by Feature Matching and MUSIC Algorithm.

Screening systems for infectious diseases based on fever have been implemented at international airports to prevent the spread of infection for over a decade. Currently, only Infrared Thermography (IRT) is used for screening and measuring facial skin temperature, which is one of clinical indicators of potential infection. Aiming at higher accuracy in screening, our group adopted heart rate (HR) and respiration rate (RR) for the first time as the new screening parameters. In our previous study, we proposed a screening system based on dual image sensors, which include IRT and a charged-coupled devices (CCD) camera. The sensors can measure three vital signs simultaneously, namely HR, RR, and facial skin temperature. For the measurement of RR in this system, stability and swiftness must be applied for application in airports. In this study, we introduce feature matching and multiple signal classification (MUSIC) algorithm in this system. Feature matching between thermal images and RGB images captured by a CCD camera and IRT, respectively, is used to detect the nose and mouth in IRT, which helps extract respiration signals corresponding to airflow from breathing. In addition, the MUSIC algorithm improves the accuracy of RR frequency estimations in limited time respiration signal and achieves swiftness. The proposed method improves stability by simultaneously detecting the nose and mouth in thermal images, and enhances the accuracy of estimated RR using the MUSIC algorithm. By using this system, we evaluate the accuracy of the estimated vital signs. The performance of this screening system was evaluated using data obtained from 12 influenza patients and 13 healthy subjects at a clinical facility in Fukushima, Japan.

Stable Contactless Sensing of Vital Signs Using RGB-Thermal Image Fusion System with Facial Tracking for Infection Screening.

Infrared thermography (IRT) has been used to screen febrile passengers in international airports for over a decade. However, fever-based infection screening using IRT suffered from low sensitivity because measurements can be affected by ambient temperature, humidity, etc. In our previous study, we proposed an RGB-thermal image fusion system to measure vital signs i.e., the RGB camera detects tiny changes in color from facial skin, associated with blood flow, to estimate heart rate, and IRT senses temperature changes around the nasal area, caused by respiration, to measure respiratory rate). The inclusion of heart and respiratory rates lead to increased screening accuracy. In the present study, to promote the widespread use of our system in real-world settings, a face detection and tracking method was developed and implemented into the system, thereby enabling the accurate and stable measurement of vital signs. We assessed heart and respiratory rate estimation via an RGB-thermal image fusion system using Bland-Altman plots and statistical analysis.

Demonstration of thermo-sensitive tetra-gel with implication for facile and versatile platform for a new class of smart gels.

A tertiary branched poly(N-isopropylacrylamide) with controlled molecular weight, distribution and the end amino-functionalization (tetra-PNIPAAm-NH2) was studied for the ability to form a gel via in situ chain-end reaction with a counterpart tertiary branched poly(ethyleneglycol) bearing N-hydroxysuccinimide end groups (tetra-PEG-NHS), a well-documented class of building block to yield the tetra-gel. Some of these polymers, both comparable and distinct (relative to the counterpart) extended chain length pairs, provided a self-standing and macroscopically homogeneous gel, which was capable of undergoing thermo-sensitive and reversible change in hydration in line with the nature of PNIPAAm. Phantom network model based calculation indicated that a half molar fraction of the polymer chains in the network remained unreacted, revealing further room for optimizing the reaction condition. Since such tetra-PNIPAAm based motif can be readily tailored to a variety of other physicochemical stimuli-responsive analogues, our finding may give important insight into a platform for 'smart' tetra-gels with exceptional mechanical properties and potentially highly controllable molecular cut-off capability.

α-Heteroarylation of ketones via the umpolung reaction of N-alkoxyenamine.

A new method has been developed for the umpolung α-heteroarylation of ketones via an N-alkoxyenamine. The treatment of ketones with tris(heteroaryl)aluminum reagents in the presence of isoxazolidine gave the corresponding α-heteroarylated ketones in moderate to good yields.

Sequential retro-ene arylation reaction of N-alkoxyenamides for the synthesis of tert-alkylamines.

A sequential retro-ene arylation reaction has been developed for the conversion of N-alkoxyenamides to the corresponding tert-alkylamines in good yields via the nucleophilic addition of a triarylaluminum reagent to an in situ generated N-acylketimine. The reaction is tolerant of a range of functional groups and provides facile access to a series of tert-alkylamines that would be otherwise difficult to access using conventional procedures.