Genetic Diagnosis of Adult Hemodialysis Patients With Unknown Etiology.

Introduction: Kidney disease of unknown etiology accounts for 1 in 10 adult end-stage renal disease (ESRD) cases worldwide. The aim of this study is to clarify the genetic background of patients with chronic kidney disease (CKD) of unknown etiology who initiated renal replacement therapy (RRT) in adulthood. Methods: This is a multicenter cross-sectional cohort study. Of the 1164 patients who attended 4 dialysis clinics in Japan, we first selected patients who started RRT between the ages of 20 and 49 years. After excluding patients with apparent causes of CKD (e.g., diabetic nephropathy, polycystic kidney disease (PKD) with family history, patients who underwent renal biopsy), 90 patients with CKD of unknown cause were included. The 298 genes associated with CKD were analyzed using capture-based targeted next-generation sequencing. Results: Of the 90 patients, 10 (11.1%) had pathogenic variants in CKD-causing genes and 17 (18.9%) had variant of unknown significance (VUS). Three patients had PKD1 pathogenic variants, and 1 patient had PKD1 and COL4A4 pathogenic variants. In addition, 2 patients were diagnosed with atypical hemolytic uremic syndrome (aHUS) due to C3 or CFHR5. One patient each was diagnosed with Alport syndrome due to COL4A4 and COL4A3 variants, nephronophthisis due to NPHP1 variants, Fabry disease due to GLA variants, and autosomal-dominant tubulointerstitial kidney disease due to UMOD variants. Genetic diagnoses were not concordant with clinical diagnoses, except for patients with PKD1 variant. Conclusion: This largest study on genetic analysis in hemodialysis-dependent adults revealed the presence of undiagnosed inherited kidney diseases.

Body fluid volume calculated using the uric acid kinetic model relates to the vascular event.

PURPOSE: We developed a method to measure the extracellular and intracellular fluid volumes using the kinetics of uric acid in the bodies of Japanese patients undergoing dialysis. In this research, we aimed to assess the prognosis of vascular events using this uric acid kinetic model method. METHODS: We conducted a retrospective cohort study of 1,298 patients who were undergoing hemodialysis or predilution online hemodiafiltration at the end of December 2019 at 13 institutions in Japan. Information on vascular events was acquired in 2020. Vascular event prognosis was defined as the new incidence of one or more of the following four types of vascular events: myocardial infarction, cerebral infarction, cerebral hemorrhage, or limb amputation. We measured the extracellular fluid volume and intracellular fluid volume after dialysis using the uric acid kinetic model method and determined the association between ECV, ICV, and vascular event risk. RESULTS: A high extracellular volume was substantially linked to an increased risk of vascular events. In addition, while a crude analysis revealed that a high intracellular volume was associated with a low risk of vascular events, this was not statistically significant after multifactorial adjustment. This result was partly affected by the low measurement accuracy of the serum urea nitrogen level used for the intracellular volume calculation. CONCLUSIONS: Extracellular volume calculated using the uric acid kinetic model method is a prognostic factor for vascular events in patients undergoing hemodialysis.

Utility of a modified Controlling Nutritional Status score in mortality risk assessment of patients on hemodialysis.

PURPOSE: Malnutrition is common among hemodialysis patients. A nutritional index may be important for their prognosis prediction. We examined the use of a modified version of the Controlling Nutritional Status (CONUT) score as a predictor of mortality in hemodialysis patients and investigated whether statin use modified the association. METHODS: We conducted a retrospective cohort study in 1130 adults (mean age 67.7 years, 35.0% women) on maintenance hemodialysis at four outpatient dialysis clinics in Japan. The exposure of interest was modified CONUT scores calculated by one-time data of serum albumin and total cholesterol at baseline, or mean data of these measures using multiple blood test results. The primary outcome was all-cause mortality. We examined the association between modified CONUT groups (low-risk, middle-risk, and high-risk) and mortality using multivariate Cox proportional hazard regression and logistic regression models in overall patients and by statin use. RESULTS: During a median follow-up of 6.6 years, 525 (46.5%) patients died. Analysis using both one-time and mean data showed increased mortality risk in the high-risk modified CONUT score group as compared to the low-risk group (mean data: HR 2.52, 95% CI 1.96, 3.24); when stratified by statin use, the effect appeared stronger among users (mean data: HR 5.84, 95% CI 2.98, 11.44). Strong predictive ability was observed, particularly for mortality risk at 5 year follow-up using mean data (AUC = 0.842). CONCLUSION: Our results suggest that the modified CONUT score may be useful in helping clinicians quickly identify hemodialysis patients with poorer prognosis who may benefit from close monitoring and interventions.

Epitaxial growth of an atom-thin layer on a LiNi0.5Mn1.5O4 cathode for stable Li-ion battery cycling.

Transition metal dissolution in cathode active material for Li-based batteries is a critical aspect that limits the cycle life of these devices. Although several approaches have been proposed to tackle this issue, this detrimental process is not yet overcome. Here, benefitting from the knowledge developed in the semiconductor research field, we apply an epitaxial method to construct an atomic wetting layer of LaTMO3 (TM = Ni, Mn) on a LiNi0.5Mn1.5O4 cathode material. Experimental measurements and theoretical analyses confirm a Stranski-Krastanov growth, where the strained wetting layer forms under thermodynamic equilibrium, and it is self-limited to monoatomic thickness due to the competition between the surface energy and the elastic energy. Being atomically thin and crystallographically connected to the spinel host lattices, the LaTMO3 wetting layer offers long-term suppression of the transition metal dissolution from the cathode without impacting its dynamics. As a result, the epitaxially-engineered cathode material enables improved cycling stability (a capacity retention of about 77% after 1000 cycles at 290 mA g-1) when tested in combination with a graphitic carbon anode and a LiPF6-based non-aqueous electrolyte solution.

Prevalence of mild cognitive impairment and its association with handgrip strength in patients on hemodialysis.

Dementia is associated with a high risk of death and hospitalization among patients on hemodialysis (HD). We retrospectively evaluated the prevalence of mild cognitive impairment (MCI) in 421 patients on maintenance HD across nine facilities and investigated whether decreased handgrip strength was associated with decreased cognitive function. The Montreal Cognitive Assessment-Japan (MoCA-J) score and handgrip strength were measured. The mean age was 69.8 ± 11.2 years, and the median dialysis vintage 74.5 (IQR 30-150) months. The median MoCA-J score was 25 (IQR 21-27), and MCI was confirmed in 245 (58.2%) patients. Both the MoCA-J score and MoCA-J executive score were associated with age, history of cerebrovascular disease (CVA), and handgrip strength after adjustments. We found, among patients on HD aged under 70 years with a history of CVA, a handgrip strength < 90% (25.2 kg in males and 16.2 kg in females) correlated with significantly lower MoCA-J scores. A high prevalence of MCI and decreased handgrip strength were observed in patients on HD. Handgrip strength may be useful for the easy detection of MCI. A decrease in handgrip strength would allow for the early detection of MCI, especially among patients on HD aged under 70 years with a history of CVA.

Effect of porous structural properties on lithium-ion and sodium-ion storage: illustrated by the example of a micro-mesoporous graphene1-x (MoS2) x anode.

In this work, we synthesized micro-mesoporous graphene1-x (MoS2) x with different compositional ratios via co-reduction of graphite oxide and exfoliated MoS2 platelets. We systematically studied the performance of the micro-mesoporous graphene1-x (MoS2) x as anodes in lithium-ion batteries and sodium-ion batteries. The results show that the specific surface areas of the composites decrease with introducing MoS2. The irreversible capacitance, which is related to the formation of solid electrolyte interphases, also decreases. Besides specific surface area, we found that micropores can benefit the lithiation and sodiation. We demonstrated that a specific charge capacity of 1319.02 mA h g-1 can be achieved at the 50th cycle for the graphene½(MoS2)½ anode in lithium-ion batteries. Possible relationships between such a high specific capacity and the micro-mesoporous structure of the graphene1-x (MoS2) x anode are discussed. This work may shed light on a general strategy for the structural design of electrode materials in lithium-ion batteries and sodium-ion batteries.

A green strategy for the preparation of a honeycomb-like silicon composite with enhanced lithium storage properties.

The high-performance silicon (Si) composite electrodes are being widely developed due to their considerable theoretical capacity. Coating with carbon-based materials is an efficient way to solve the common issues of Si-based materials. Currently, most of the reported strategies are complicated, pollutive, or uneconomic, which hamper their practical applications. Herein, a honeycomb-like Si-based composite was prepared to address these issues via a facile and green reduction approach at room temperature. The pre-anchored Si nanoparticles could be packed and interconnected through a three-dimensional graphene network to further enhance the electrochemical properties of the active materials. As an electrode, this composite shows good rate capabilities upon lithium storage and cycling stability. The continued cycling measurement delivers a -0.049% capacity decay rate per cycle within 600 cycles. A direct comparison further exhibits the obviously improved performance between the as-designed Si-based composite and naked Si, suggesting a potential application of this convenient strategy for other high-performance electrode materials.

Prevalence of rear seat belt use among pregnant women in a suburban area of Japan.

AIM: The aim of this study was to clarify the prevalence and influencing factors of rear seat belt use among pregnant women. METHODS: Questionnaires were given to 1546 pregnant women who visited obstetrics clinics and hospitals for prenatal checkups from October to December 2013. A total of 1494 pregnant women (96.6%) agreed to participate in this study and completed the questionnaire. RESULTS: Fewer than 20% of the rear-seat passengers 'always' used seat belts before and during pregnancy, whereas a third 'never' used a seat belt before or during pregnancy. There was no significant decrease in seat belt use by rear-seat passengers during compared to before pregnancy. Multivariate analysis revealed that age, knowledge of how to use a seat belt during pregnancy, belief in the compulsory use of a rear seat belt and driver behavioral characteristics before pregnancy were associated with rear seat belt use during pregnancy. CONCLUSIONS: The prevalence of fastening seat belts was substantially low. The provision of information regarding proper seat belt use and its role in protecting the fetus may increase use.

Comparison of reduction products from graphite oxide and graphene oxide for anode applications in lithium-ion batteries and sodium-ion batteries.

Hydrazine-reduced graphite oxide and graphene oxide were synthesized to compare their performances as anode materials in lithium-ion batteries and sodium-ion batteries. Reduced graphite oxide inherits the layer structure of graphite, with an average spacing between neighboring layers (d-spacing) of 0.374 nm; this exceeds the d-spacing of graphite (0.335 nm). The larger d-spacing provides wider channels for transporting lithium ions and sodium ions in the material. We showed that reduced graphite oxide as an anode in lithium-ion batteries can reach a specific capacity of 917 mA h g-1, which is about three times of 372 mA h g-1, the value expected for the LiC6 structures on the electrode. This increase is consistent with the wider d-spacing, which enhances lithium intercalation and de-intercalation on the electrodes. The electrochemical performance of the lithium-ion batteries and sodium-ion batteries with reduced graphite oxide anodes show a noticeable improvement compared to those with reduced graphene oxide anodes. This improvement indicates that reduced graphite oxide, with larger interlayer spacing, has fewer defects and is thus more stable. In summary, we found that reduced graphite oxide may be a more favorable form of graphene for the fabrication of electrodes for lithium-ion and sodium-ion batteries and other energy storage devices.

Understanding Performance Differences from Various Synthesis Methods: A Case Study of Spinel LiCr0.2Ni0.4Mn1.4O4 Cathode Material.

High voltage (5-V class) spinel LiCr0.2Ni0.4Mn1.4O4 is one of the most promising cathode materials to meet the energy requirements of lithium-ion batteries for electric vehicles and hybrid electric vehicles. For the mass production of this material (1 kg or higher), different synthesis routes will lead to different electrochemical performances, even with similar morphology and similar crystal structure obtained from laboratory X-ray diffraction, and the reason for this issue is still not clear. Herein, we have investigated the reasons for the different electrochemical performances resulting from three common synthesis routes (spray pyrolysis, coprecipitation, and sol-gel). Taking advantage of the high-resolution X-ray beam in synchrotron X-ray diffraction, we find that varying phase composition and the generated impurities, rather than the particle distribution, are likely to be the main reasons for the detected electrochemical variations. A higher amount of impurities will result in greater charge transfer resistance, inferior cycling stability, and more oxygen/lithium vacancies. Therefore, it is very important to obtain a deeper understanding with the help of higher-resolution X-rays and to provide better guidance for mass production of this cathode material for practical applications.

Enhancing the high rate capability and cycling stability of LiMn2O4 by coating of solid-state electrolyte LiNbO3.

To study the influence of solid-state electrolyte coating layers on the performance of cathode materials for lithium-ion batteries in combination with organic liquid electrolyte, LiNbO3-coated Li1.08Mn1.92O4 cathode materials were synthesized by using a facile solid-state reaction method. The 0.06LiNbO3-0.97Li1.08Mn1.92O4 cathode exhibited an initial discharge capacity of 125 mAh g(-1), retaining a capacity of 119 mAh g(-1) at 25 °C, while at 55 °C, it exhibited an initial discharge capacity of 130 mAh g(-1), retaining a capacity of 111 mAh g(-1), both at a current density of 0.5 C (where 1 C is 148 mAh g(-1)). Very good rate capability was demonstrated, with the 0.06LiNbO3-0.97Li1.08Mn1.92O4 cathode showing more than 85% capacity at the rate of 50 C compared with the capacity at 0.5 C. The 0.06LiNbO3-0.97Li1.08Mn1.92O4 cathode showed a high lithium diffusion coefficient (1.6 × 10(-10) cm(2) s(-1) at 55 °C), and low apparent activation energy (36.9 kJ mol(-1)). The solid-state electrolyte coating layer is effective for preventing Mn dissolution and maintaining the high ionic conductivity between the electrode and the organic liquid electrolyte, which may improve the design and construction of next-generation large-scale lithium-ion batteries with high power and safety.

Novel germanium/polypyrrole composite for high power lithium-ion batteries.

Nano-Germanium/polypyrrole composite has been synthesized by chemical reduction method in aqueous solution. The Ge nanoparticles were directly coated on the surface of the polypyrrole. The morphology and structural properties of samples were determined by X-ray diffraction, scanning electron microscopy and transmission electron microscopy. Thermogravimetric analysis was carried out to determine the polypyrrole content. The electrochemical properties of the samples have been investigated and their suitability as anode materials for the lithium-ion battery was examined. The discharge capacity of the Ge nanoparticles calculated in the Ge-polypyrrole composite is 1014 mAh g(-1) after 50 cycles at 0.2 C rate, which is much higher than that of pristine germanium (439 mAh g(-1)). The composite also demonstrates high specific discharge capacities at different current rates (1318, 1032, 661, and 460 mAh g(-1) at 0.5, 1.0, 2.0, and 4.0 C, respectively). The superior electrochemical performance of Ge-polypyrrole composite could be attributed to the polypyrrole core, which provides an efficient transport pathway for electrons. SEM images of the electrodes have demonstrated that polypyrrole can also act as a conductive binder and alleviate the pulverization of electrode caused by the huge volume changes of the nanosized germanium particles during Li(+) intercalation/de-intercalation.

Interface strain-induced multiferroicity in a SmFeO3 film.

An epitaxial pseudocubic SmFeO3 thin film on (100) Nb-SrTiO3 was studied based on ferroelectric (FE) characterization and magnetic measurements. High-resolution transmission electron microscopy images clarify the nature of the epitaxial growth, the stress-induced structural distortion at the film/substrate interface, and the existence of two different orientation lattices. Clear grain boundaries can be seen, which could introduce an extra local distortion. Rectangular FE loops can be observed at room temperature, even by just applying a small voltage ranging from -1 to +1 V, indicative of the presence of FE polarization. Piezoelectric force microscopy images confirm the existence of FE domains and the switchable polarization. A strong ferromagnetic-like transition occurs around 185 K, which is much lower than the transition observed in the bulk sample. It is believed that the pseudocubic structure enhances FE polarization and decreases the magnetic ordering temperature, which is confirmed by the first-principles theoretical calculations. Meanwhile, the ferroelectricity in this thin film should originate from distortion and modification in the structural modules rather than from the exchange striction interaction that is found in the bulk SmFeO3.

Preparation of multiple site-specific mutant proteins for NMR studies by PCR-directed cell-free protein synthesis.

Cell-free protein synthesis (CFPS) offers a fast and inexpensive approach to selectively label proteins with isotopes that can then be detected by nuclear magnetic resonance (NMR) spectroscopy directly in the translation mixture. We describe a PCR-based approach for production of protein-coding circularized DNA templates that can be expressed in Escherichia coli extract in CFPS dialysis system. This approach typically yields target protein concentrations close to 1 mg/mL, which is sufficient for subsequent analysis by 2D (1)H,(15)N-NMR. Furthermore, this PCR-based technique also enables parallel preparation of mutant proteins in a high-throughput mode, enabling rapid assignments of NMR signals. This chapter describes the general CFPS protocol that we used to rapidly assign residue-specific cross peaks from 2D (1)H,(15 N-NMR) spectra obtained from 12 Ile/Ala substituted mutants of the 40 kDa protein complex, αCTS:τc16.

Site-specific incorporation of unnatural amino acids into proteins by cell-free protein synthesis.

Cell-free protein synthesis (CFPS) offers a fast and inexpensive means to incorporate unnatural amino acids (UAAs) site specifically into proteins. This enables engineering of proteins and allows production of protein-based probes for analysis of their interactions with other molecules. Using dialysis Escherichia coli CFPS system in combination with aminoacyl-tRNA synthetase and suppressor tRNA evolved from Methanocaldococcus jannaschii high expression yield of proteins with site specifically incorporated UAAs can be achieved. Typically the target protein can be prepared at concentrations of about 1 mg/mL, which is generally sufficient for subsequent applications.

Hollow structured Li3VO4 wrapped with graphene nanosheets in situ prepared by a one-pot template-free method as an anode for lithium-ion batteries.

To explore good anode materials of high safety, high reversible capacity, good cycling, and excellent rate capability, a Li3VO4 microbox with wall thickness of 40 nm was prepared by a one-pot and template-free in situ hydrothermal method. In addition, its composite with graphene nanosheets of about six layers of graphene was achieved. Both of them, especially the Li3VO4/graphene nanosheets composite, show superior electrochemical performance to the formerly reported vanadium-based anode materials. The composite shows a reversible capacity of 223 mAh g(-1) even at 20C (1C = 400 mAh g(-1)). After 500 cycles at 10C there is no evident capacity fading.

Proofreading exonuclease on a tether: the complex between the E. coli DNA polymerase III subunits α, epsilon, θ and ß reveals a highly flexible arrangement of the proofreading domain.

A complex of the three (αεθ) core subunits and the ß2 sliding clamp is responsible for DNA synthesis by Pol III, the Escherichia coli chromosomal DNA replicase. The 1.7 Å crystal structure of a complex between the PHP domain of α (polymerase) and the C-terminal segment of ε (proofreading exonuclease) subunits shows that ε is attached to α at a site far from the polymerase active site. Both α and ε contain clamp-binding motifs (CBMs) that interact simultaneously with ß2 in the polymerization mode of DNA replication by Pol III. Strengthening of both CBMs enables isolation of stable αεθ:ß2 complexes. Nuclear magnetic resonance experiments with reconstituted αεθ:ß2 demonstrate retention of high mobility of a segment of 22 residues in the linker that connects the exonuclease domain of ε with its α-binding segment. In spite of this, small-angle X-ray scattering data show that the isolated complex with strengthened CBMs has a compact, but still flexible, structure. Photo-crosslinking with p-benzoyl-L-phenylalanine incorporated at different sites in the α-PHP domain confirm the conformational variability of the tether. Structural models of the αεθ:ß2 replicase complex with primer-template DNA combine all available structural data.

A direct proofreader-clamp interaction stabilizes the Pol III replicase in the polymerization mode.

Processive DNA synthesis by the αεθ core of the Escherichia coli Pol III replicase requires it to be bound to the ß2 clamp via a site in the α polymerase subunit. How the ε proofreading exonuclease subunit influences DNA synthesis by α was not previously understood. In this work, bulk assays of DNA replication were used to uncover a non-proofreading activity of ε. Combination of mutagenesis with biophysical studies and single-molecule leading-strand replication assays traced this activity to a novel ß-binding site in ε that, in conjunction with the site in α, maintains a closed state of the αεθ-ß2 replicase in the polymerization mode of DNA synthesis. The ε-ß interaction, selected during evolution to be weak and thus suited for transient disruption to enable access of alternate polymerases and other clamp binding proteins, therefore makes an important contribution to the network of protein-protein interactions that finely tune stability of the replicase on the DNA template in its various conformational states.

Lanthanide tags for site-specific ligation to an unnatural amino acid and generation of pseudocontact shifts in proteins.

Pseudocontact shifts (PCS) from paramagnetic lanthanide ions present powerful long-range structural restraints for structural biology by NMR spectroscopy, but site-specific tagging of proteins with lanthanides remains a challenge, as most of the available lanthanide tags require proteins with single cysteine residues. We show that cyclen-based paramagnetic lanthanide tags can be attached to proteins in a site-specific manner by Cu(I)-catalyzed azide-alkyne cycloaddition to a genetically encoded p-azido-l-phenylalanine residue with a tether that proved sufficiently short and rigid for the observation of PCSs in several proteins. Despite the sterically demanding conditions associated with bulky tags and reactions close to the protein surface, ligation yields consistently above 50% and approaching 100% were obtained with the help of the Cu(I)-stabilizing ligand BTTAA. The yields were high independent of the presence of cysteine residues, thereby avoiding the need for cysteine mutations associated with conventional lanthanide-tagging strategies.