Method for quantifying free hemoglobin, distinct from the hemoglobin-haptoglobin complex, in human serum.

The measurement of free hemoglobin (free Hb) in blood is crucial for assessing the risk of organ damage in patients with hemolytic diseases. However, the colorimetric method, commonly used in clinical practice, does not distinguish between free Hb and the hemoglobin-haptoglobin complex (Hb-Hp) in the blood, instead reflecting the total Hb level. Although size-exclusion high-performance liquid chromatography (SEC-HPLC) can specifically measure free Hb, its clinical use is limited by long assay times. Here, we developed a novel assay method for the rapid quantification of free Hb in serum, distinguishing it from Hb-Hp, using a latex agglutination immunoturbidimetric assay (LATIA). This method could be used to measure free Hb in sera in the range of 1-100 µg/mL in approximately 15 min using an automatic biochemistry analyzer. Using Hb-spiked serum samples from healthy adults, there was a high correlation with Hb levels determined using the newly developed method and SEC-HPLC, indicating a high specificity for free Hb. This novel assay can be used to monitor levels of free Hb in patients with various hemolytic diseases and to design therapeutic strategies based on measured values. However, further studies are required to assess its clinical performance.

Dysfunction of Foxp3+ Regulatory T Cells Induces Dysbiosis of Gut Microbiota via Aberrant Binding of Immunoglobulins to Microbes in the Intestinal Lumen.

Foxp3+ regulatory T (Treg) cells prevent excessive immune responses against dietary antigens and commensal bacteria in the intestine. Moreover, Treg cells contribute to the establishment of a symbiotic relationship between the host and gut microbes, partly through immunoglobulin A. However, the mechanism by which Treg cell dysfunction disturbs the balanced intestinal microbiota remains unclear. In this study, we used Foxp3 conditional knockout mice to conditionally ablate the Foxp3 gene in adult mice and examine the relationship between Treg cells and intestinal bacterial communities. Deletion of Foxp3 reduced the relative abundance of Clostridia, suggesting that Treg cells have a role in maintaining Treg-inducing microbes. Additionally, the knockout increased the levels of fecal immunoglobulins and immunoglobulin-coated bacteria. This increase was due to immunoglobulin leakage into the gut lumen as a result of loss of mucosal integrity, which is dependent on the gut microbiota. Our findings suggest that Treg cell dysfunction leads to gut dysbiosis via aberrant antibody binding to the intestinal microbes.

Fabrication of atomic junctions with experimental parameters optimized using ground-state searches of Ising spin computing.

Feedback-controlled electromigration (FCE) is employed to control metal nanowires with quantized conductance and create nanogaps and atomic junctions. In the FCE method, the experimental parameters are commonly selected based on experience. However, optimization of the parameters by way of tuning is intractable because of the impossibility of attempting all different combinations systematically. Therefore, we propose the use of the Ising spin model to optimize the FCE parameters, because this approach can search for a global optimum in a multidimensional solution space within a short calculation time. The FCE parameters were determined by using the energy convergence properties of the Ising spin model. We tested these parameters in actual FCE experiments, and we demonstrated that the Ising spin model could improve the controllability of the quantized conductance in atomic junctions. This result implies that the proposed method is an effective tool for the optimization of the FCE process in which an intelligent machine can conduct the research instead of humans.

Control parameters for fabrication of single-electron transistors using field-emission-induced electromigration.

We report a simple method for the control of electrical characteristics of planar-type metal-based single-electron transistors (SETs) using field-emission-induced electromigration. The advantages of this method are as follows: (1) the fabrication of SETs is achieved by only passing a field emission current through a nanogap and (2) the charging energy of SETs can be controlled by adjusting the magnitude of the applied current during the procedure. In order to better control the electrical properties of the SETs, we investigate the relation between control parameters of the method and electrical characteristics of the SETs. When the field-emission-induced electromigration with the preset current of 500 nA was applied to the nanogaps, current-voltage characteristics of the nanogaps displayed the suppression of electrical current at low-bias voltages known as Coulomb blockade at 16 K. In addition, Coulomb blockade voltage was clearly modulated by the gate voltage periodically at 16 K, resulting in the formation of single island in the SETs by the field-emission-induced electromigration. Furthermore, as the preset current was increased, the charging energy of the SETs was decreased with decreasing the initial gap separation of the nanogaps. These results imply that the electrical characteristics of the SETs are controllable by the preset current of the method and the initial gap separation of the nanogaps. Field-emission-induced electromigration procedure allows us to simply control electrical characteristics of planar-type metal-based SETs.

Bioluminescent enzyme immunoassay for the detection of norovirus capsid antigen.

An ultrasensitive and fully automated bioluminescent enzyme immunoassay (BLEIA) was developed for the detection of norovirus (NV) capsid antigen. In the evaluation tests with recombinant virus-like particles, the BLEIA demonstrated broad reactivity against several NV genotypes (genotypes 1, 3, 4, 7, 8, and 12 in genogroup I [GI] and genotypes 1, 2, 3, 4, 5, 6, 12, and 13 in GII), a wide dose-response range from 0.25 pg/ml to 10,000 pg/ml, and good reproducibility with low coefficients of variation (CVs) (within-run CVs of <2.8%, between-day CVs of <3.7%). In the evaluation tests with NV-positive fecal samples, a good correlation (y = 0.66x - 3.21, r = 0.84) between the BLEIA and real-time quantitative reverse transcription-PCR was obtained. Furthermore, in the dilution test with NV specimens, the analytical sensitivity of NV was estimated to be 10(5) to 10(6) copies/g of fecal sample, indicating that the analytical sensitivity of the BLEIA is comparable to that of commercially available molecular methods. All assay steps are fully automated, the turnaround time is 46 min, and the throughput of the assay is 120 tests/h. These results indicate that the BLEIA is potentially useful for the rapid diagnosis of NV in epidemic and sporadic gastroenteritis.