Transcriptomic profiling and the first spatial expression analysis of candidate genes in the salivary gland of the East Asian medicinal leech, Hirudo nipponia.

Hirudo nipponia, a blood-sucking leech native to East Asia, possesses a rich repertoire of active ingredients in its saliva, showcasing significant medical potential due to its anticoagulant, anti-inflammatory, and antibacterial effects against human diseases. Despite previous studies on the transcriptomic and proteomic characteristics of leech saliva, which have identified medicinal compounds, our knowledge of tissue-specific transcriptomes and their spatial expression patterns remains incomplete. In this study, we conducted an extensive transcriptomic profiling of the salivary gland tissue in H. nipponia based on de novo assemblies of tissue-specific transcriptomes from the salivary gland, teeth, and general head region. Through gene ontology (GO) analysis and hierarchical clustering, we discovered a novel set of anti-coagulant factors-i.e., Hni-Antistasin, Hni-Ghilanten, Hni-Bdellin, Hni-Hirudin-as well as a previously unrecognized immune-related gene, Hni-GLIPR1 and uncharacterized salivary gland specific transcripts. By employing in situ hybridization, we provided the first visualization of gene expression sites within the salivary gland of H. nipponia. Our findings expand on our understanding of transcripts specifically expressed in the salivary gland of blood-sucking leeches, offering valuable resources for the exploration of previously unidentified substances with medicinal applications.

In-air experimental achievement of high-voltage DC power supply to be installed in a tandem electrostatic accelerator system for boron neutron capture therapy application.

The boron neutron capture therapy (BNCT) system developed by the Korea Institute of Radiological and Medical Sciences is a compact neutron source that can be installed at medical institutes. The target energy was accelerated to a maximum of 2.4 MeV-20 mA by introducing a gas stripping device that converts negative hydrogen ions into positive ions. By using the tandem-type accelerator in this way, a high-voltage DC power supply was designed with 1.2 MV-45 mA as the maximum capability. The design was improved to reduce the number of stages of a Cockcroft-Walton voltage multiplier. Hence, the ripple risk of the DC flat top resulting from unwanted stray capacitance was lowered. The overall height and volume of the Cockcroft-Walton voltage multiplier were reduced to less than half those of the existing design method, making miniaturization possible. After such advanced design and manufacturing, performance tests were performed at 750 kV-45 mA under 23 stages of the Cockcroft-Walton voltage multiplier, which is the highest level that can perform at its maximum under in-air conditions. It demonstrated stable performance under in-air conditions without breakdown for 2 h, even at 620 kV-35 mA. To reach the final target of 1.2 MV-45 mA, the groundwork is laid for achieving experimental performance while satisfying the optimal requirements in SF6 gas.

Optimal design of high-voltage DC power supply of 1.2 MV/45 mA applied to boron neutron capture therapy system.

To build a proton beam accelerator that can be applied to a boron neutron capture therapy system based on an electrostatic accelerator, a high-voltage direct-current (DC) power supply system equivalent to the generation of neutrons should be provided. The symmetrical Cockcroft-Walton voltage multiplier method is suitable for stable acceleration of the proton beam in the tandem electrostatic accelerator in this system. Before the second step-up with the Cockcroft-Walton circuit, the design of the inverter is prioritized by preponderantly considering the first voltage and resonance frequency. Moreover, the optimized stacking number is determined with consideration of the ripple voltage, voltage drop, average output voltage, and fundamental harmonics, and a design is performed to set related parameter values to be stable in the flat-top region of the voltage. A high-voltage DC power supply system of 1.2 MV/45 mA is needed for a stable terminal energy of 2.4 MeV/20 mA. Such a design can be optimized by securing reliable data using a simulation tool on the basis of theoretical calculations. This will become a formidable touchstone in manufacturing technology based on acquiring practical know-how for setting up a tandem electrostatic accelerator-based boron neutron capture therapy system in the future.

A Rapid Assessing Method of Drug Susceptibility Using Flow Cytometry for Mycobacterium tuberculosis Isolates Resistant to Isoniazid, Rifampin, and Ethambutol / 결핵및호흡기질환

Background@#The current conventional drug susceptibility test (DST) for Mycobacterium tuberculosis (Mtb) takes several weeks of incubation to obtain results. As a rapid method, molecular DST requires only a few days to get the results but does not fully cover the phenotypic resistance. A new rapid method based on the ability of viable Mtb bacilli to hydrolyze fluorescein diacetate to free fluorescein with detection of fluorescent mycobacteria by flow cytometric analysis, was recently developed. @*Methods@#To evaluate this cytometric method, we tested 39 clinical isolates which were susceptible or resistant to isoniazid (INH) or rifampin (RIF), or ethambutol (EMB) by phenotypic or molecular DST methods and compared the results. @*Results@#The susceptibility was determined by measuring the viability rate of Mtb and all the isolates which were tested with INH, RIF, and EMB showed susceptibility results concordant with those by the phenotypic solid and liquid media methods. The isolates having no mutations in the molecular DST but resistance in the conventional phenotypic DST were also resistant in this cytometric method. These results suggest that the flow cytometric DST method is faster than conventional agar phenotypic DST and may complement the results of molecular DST. @*Conclusion@#In conclusion, the cytometric method could provide quick and more accurate information that would help clinicians to choose more effective drugs.

Performance of an impedance-variable pulsed high-power electron-beam accelerator based on energy efficient transmission.

Versatile high-power pulsed electron-beam accelerators that meet the requirements of pulsed high-power specifications are needed for appropriate applications in medical industry, defense, and other industries. The pulsed electron beam accelerator comprising a Marx generator and Blumlein pulse forming line (PFL) is designed to accelerate the electron beams at the level of 1 MeV when electrostatically discharging. The performance specifications of Marx generators consisting of a 100 kV DC power supply, R-L-C circuit, and high voltage switch are at a maximum 800 kV. At this time, by using the capacitance mismatching principle between the Marx generator and the Blumlein PFL under the law of preserving the amount of charge, it is possible to generate a high voltage in the form of a square pulse up to about 1.1 MV, as much as 1.37 times the charged voltage of the Marx generator. As a result, energy transmission from the Marx generator with a high efficiency of about 85% to the Blumlein PFL is possible. The aim of this study is that the pulsed high-power electron-beam accelerator can be used to change the diode impedance, and the energy of the accelerated electron beam reaches a level of 1 MeV with the square pulse width of about 100 ns at the flat-top in the range of relativistic electron beam generation. Performance tests were securely carried out by installing a dummy load based on CuSO4 solution varying the diode impedance to deter damage to the circuit by preventing reflected waves from being generated in the load.

Evaluation of Auricular Cartilage Reconstruction Using a 3-Dimensional Printed Biodegradable Scaffold and Autogenous Minced Auricular Cartilage.

Auricular cartilage reconstruction represents one of the greatest challenges for otolaryngology-head and neck surgery. The native structure and composition of the auricular cartilage can be achieved by combining a suitable chondrogenic cell source with an appropriate scaffold. In reconstructive surgery for cartilage tissue, autogenous cartilage is considered to be the best chondrogenic cell source. Polycaprolactone is mainly used as a tissue-engineered scaffold owing to its mechanical properties, miscibility with a large range of other polymers, and biodegradability. In this study, scaffolds with or without autogenous minced auricular cartilage were implanted bilaterally in rabbits for auricular regeneration. Six weeks (n = 4) and 16 weeks (n = 4) after implantation, real-time quantitative reverse transcription polymerase chain reaction and histology were used to assess the regeneration of the auricular cartilage. Quantitative reverse transcription polymerase chain reaction analysis revealed that the messenger RNA expression of aggrecan, collagen I, and collagen II was higher in scaffolds with 50% minced cartilage than the scaffold-only groups or scaffolds with 30% minced cartilage (P < 0.05). Furthermore, histological analysis demonstrated significantly superior cartilage regeneration in scaffolds with the minced cartilage group compared with the scaffold-only and control groups (P < 0.05). Autogenous cartilage can be easily obtained and loaded onto a scaffold to promote the presence of chondrogenic cells, allowing for an improvement of the reconstruction of auricular cartilage. Here, the regeneration of auricular cartilage was also successful in the 50% minced cartilage group. The results presented in this study could have clinical implications, as they demonstrate the potential of a 1-stage process for auricular reconstruction.

Miniaturized two-stack Blumlein pulser with a variable repetition-rate for non-thermal irreversible-electroporation experiments.

Non-thermal irreversible electroporation (NTIRE) to avoid thermal damage to cells during intense DC ns pulsed electric fields (nsPEFs) is a recent modality for medical applications. This mechanism, related to bioelectrical dynamics of the cell, is linked to the effect of a DC electric field and a threshold effect with an electrically stimulated membrane for the charge distribution in the cell. To create the NTIRE condition, the pulse width of the nsPEF should be shorter than the charging time constant of the membrane related to the cell radius, membrane capacitance, cytoplasm resistivity, and medium resistivity. It is necessary to design and fabricate a very intense nanosecond DC electric field pulser that is capable of producing voltages up to the level of 100 kV/cm with an artificial pulse width (∼ns) with controllable repetition rates. Many devices to generate intense DC nsPEF using various pulse-forming line technologies have been introduced thus far. However, the previous Blumlein pulse-generating devices are clearly inefficient due to the energy loss between the input voltage and the output voltage. An improved two-stage stacked Blumlein pulse-forming line can overcome this limitation and decrease the energy loss from a DC power supply. A metal oxide silicon field-effect transistor switch with a fast rise and fall time would enable a high repetition rate (max. 100 kHz) and good endurance against very high voltages (DC ∼ 30 kV). The load is designed to match the sample for exposure to cell suspensions consisting of a 200 Ω resistor matched with a Blumlein circuit and two electrodes without the characteristic RC time effect of the circuit (capacitance =0.174 pF).

Potential Role of Immunodiagnosis for Pulmonary Tuberculosis Using Induced Sputum Cells

PURPOSE: To evaluate the diagnostic utility and predictors for determinate results of an enzyme-linked immunospot assay using induced sputum cells (IS ELISPOT) for a rapid diagnosis of pulmonary tuberculosis (TB). MATERIALS AND METHODS: Subjects suspected of pulmonary TB who had either sputum acid fast bacilli smear-negative or not producing sputum spontaneously were prospectively enrolled. ELISPOT assay was performed using cells from induced sputum. RESULTS: A total of 43 subjects, including 25 with TB (TB group) and 18 with non-TB disease (non-TB group) were enrolled. Results of IS ELISPOT were determinate in only 17/43 (39%) subjects, but all of determinate results were consistent with the final diagnosis. Of the 43 sputum samples, 11 (26%) were inadequate to perform IS ELISPOT. Of 32 adequate sputum samples, the proportion of determinate results was significantly higher in the TB group (75%, 15/20) than in the non-TB group (17%, 2/12) (p=0.002). The status of active TB was a unique predictor but smear positivity was not a significant predictor for determinate results. In addition, sensitivity of IS ELISPOT (75%, 9/12) in smear negative TB was higher than that of TB-polymerase chain reaction (25%, 3/12). CONCLUSION: IS ELISPOT showed relatively high diagnostic value and accuracy in the TB group, independent of smear positivity. IS ELISPOT may provide additional diagnostic yield for microbiological tools in the rapid diagnosis of smear-negative TB.

Surface-coupling of Cerenkov radiation from a modified metallic metamaterial slab via Brillouin-band folding.

Metallic metamaterials with positive dielectric responses are promising as an alternative to dielectrics for the generation of Cerenkov radiation [J.-K. So et al., Appl. Phys. Lett. 97(15), 151107 (2010)]. We propose here by theoretical analysis a mechanism to couple out Cerenkov radiation from the slab surfaces in the transverse direction. The proposed method based on Brillouin-zone folding is to periodically modify the thickness of the metamaterial slab in the axial direction. Moreover, the intensity of the surface-coupled radiation by this mechanism shows an order-of-magnitude enhancement compared to that of ordinary Smith-Purcell radiation.

A new method to measure the polymerization shrinkage kinetics of composites using a particle tracking method with computer vision.

OBJECTIVES: The aim of this study was to develop a new method to measure the polymerization shrinkage of light cured composites and to evaluate the overall utility and significance of the technique. METHODS: An optical instrument to measure the linear polymerization shrinkage of composites without directly contacting the specimen was developed using a particle tracking method with computer vision. The measurement system consisted of a CCD color video camera, a lens, an image storage device, and image processing and analysis software. The shrinkage kinetics of a commercial silorane-based composite (P90) and two conventional methacrylate-based composites (Z250 and a flowable Z350) were investigated and compared with the data measured using the "bonded disc method". RESULTS: The linear shrinkage of the composites was 0.33-1.41%. The shrinkage value was lowest for the silorane-based (P90) composite and highest for the flowable Z350 composite. The estimated volume shrinkages of the materials were comparable to the axial shrinkages measured with the bonded disc method. SIGNIFICANCE: The new instrument was able to measure the true linear shrinkage of composites without sensitivity to the specimen geometry and the viscosity of the material. Therefore, this instrument can be used to characterize the shrinkage kinetics for a wide range of commercial and experimental visible-light-cure materials in relation to the composition and chemistry.

Enhanced transmission of electromagnetic waves through 1D plasmonic crystals.

Transmission of electromagnetic waves through thick perfect conducting slabs perforated by one-dimensional arrays of rectangular holes was studied experimentally in the microwave frequency range. The observed thickness-dependent transmission clearly exhibits the evanescent and propagating nature of the involved electromagnetic excitations on the considered structures, which are effective surface plasmons and localized waveguide resonances, respectively. The 1D crystals showing transmission based on localized resonances further manifests the frequency-dependent effective refractive index depending on the filling ratio of the holes and accompanies resonant guided wave propagation.

Effect of shrinkage strain, modulus, and instrument compliance on polymerization shrinkage stress of light-cured composites during the initial curing stage.

OBJECTIVES: The aim of this study was to investigate the influence of shrinkage strain, modulus, and instrument compliance on the polymerization shrinkage stress measurement of light-cured composites, and to determine whether the silorane-based low-shrinkage composite shows a low-polymerization shrinkage stress. METHODS: A universal hybrid; Z250 (Z2), a flowable; Z350 (Z3), and a silorane-based; P90 (P9) composite was examined. A modified "bonded disc method" was used to measure the axial shrinkage strain of the composite. For the measurement of the initial modulus development of composites during light curing, a dynamic oscillatory shear test was undertaken using a custom-made oscillation rheometer. A frequency of 6 Hz and strain amplitude of 0.0091 rad for 20 s was employed and the complex shear modulus (G(*)) was determined. A newly designed stress-strain analyzer was made to measure the shrinkage stress of the composites with two modes: (1) high compliance, or (2) low compliance. The shift between the two modes was controlled by an On-Off switch of a negative feedback circuit. Theoretical shrinkage stress was calculated from the shrinkage strain and modulus measured above, and compared with experimentally measured stress. Data were analyzed with one-way ANOVA and Tukey's post hoc test (alpha=0.05), and correlation analysis was done to investigate the relationship between measured stress and shrinkage strain, modulus, and theoretical stress. RESULTS: The shrinkage strain of Z3 (4.12%) at 10 min was the highest, followed by Z2 (2.31%) and P9 (0.77%). At 10 s after light curing, Z2 showed the highest modulus (466.2 MPa), Z3 (154.1 MPa), and P9 the lowest (130.7 MPa). The measured stresses with low compliance were much higher than those with high compliance. With high compliance, the contraction stress of Z3 was the highest (2.75 MPa), followed by Z2 (1.54 MPa) and P9 (0.48 MPa). In low-compliance mode, the stresses of Z3 (7.93 MPa) and Z2 (7.48 MPa) were similar (p=0.323) while the stress of P9 (3.23 MPa) was much lower. A strong correlation was observed between the theoretical stress and the measured stress with low compliance (R=0.996). In high-compliance mode, the shrinkage strain also showed a near-linear relationship with the stress measured (R=0.937), but the modulus showed a low correlation with the measured stress (R=0.398). SIGNIFICANCE: Depending on the instrument compliance, polymerization shrinkage stress showed significant differences for each material. In high-compliance shrinkage strain played a greater role, while in low-compliance shrinkage strain and elastic modulus contributed comparably in determining the shrinkage stress. The low-shrinkage silorane-based composite demonstrated considerable reduction in shrinkage strain and stress.

Slumping tendency and rheological properties of flowable composites.

OBJECTIVES: The aim of this study was to develop a method for measuring the slumping tendency of flowable resin composites and to correlate the results with those obtained from standard rheological methods. METHODS: Five commercial flowable composites (Aeliteflo: AF, Filtek flow: FF, DenFil flow: DF, Tetric flow: TF and Revolution: RV) were used. A fixed volume of each composite was extruded from a syringe onto a glass slide using a custom-made loading device. The composite was allowed to slump for 10s at 25 degrees C and light cured. The aspect ratio (height/diameter) of the cone or dome shaped specimen was measured to estimate the slumping tendency of the composites. In order to investigate the relationship between the slumping tendency and the rheological properties of the composites, the complex viscosity eta* of each composite was measured by a dynamic oscillatory shear test over a range of angular frequency omega=0.1-100rad/s using a rheometer. The aspect ratios of the composites were compared by one-way ANOVA and Tukey's post hoc test at the 5% significance level. Regression analysis was performed to investigate the relationship between the aspect ratio and the complex viscosity. RESULTS: Slumping tendency based on the aspect ratio varied among the five materials (AF

Initial dynamic viscoelasticity change of composites during light curing.

OBJECTIVES: The aim of this study was to measure the initial dynamic viscoelastic modulus change of dental composites during light curing using a custom made oscillation rheometer. METHODS: Six commercial universal hybrid resin composites: Z100 (Z1), Z250 (Z2), Z350 (Z3), DenFil (DF), Tetric Ceram (TC), and Clearfil AP-X (CF) were examined. A custom designed oscillation rheometer was made, which consisted of three parts: (1) an oscillatory shear strain induction unit, (2) a measuring unit of parallel plates made of glass rods, and (3) a stress measurement unit. For the measurement of the initial viscoelastic modulus change of composites during light curing as a function of time, a dynamic oscillatory shear test was undertaken at a frequency of 6Hz and strain amplitude of 0.00579rad for 10s. From the strain-stress curves, the complex shear modulus (G*), storage shear modulus (G'), loss shear modulus (G'') and loss tangent (tandelta), and the time to reach a G* of 10MPa were determined. The data were analyzed with one-way ANOVA and Tukey's post hoc test (alpha=0.05). RESULTS: There were great differences in the development of viscoelasticity among materials. At 10s, the complex modulus G* of Z1 was the highest (563.7MPa), followed by CF, Z2, Z3, TC, and finally DF being the lowest (150.3MPa). The time to reach the G* of 10MPa ranged from 2.55s (Z1) to 4.06s (DF). The tandelta of the composites decreased with time as the polymerization reaction proceeded. SIGNIFICANCE: The initial modulus development of composites during curing plays a very important role in determining the polymerization shrinkage stress. The custom made oscillation rheometer was able to measure the very initial dynamic viscoelasticity change of several commercial composites during light curing; therefore, the instrument can be used to study the initial curing kinetics of newly developed composites.