Weissella koreensis and Pediococcus pentosaceus bacterial ghosts induce inflammatory responses as immunostimulants.

In this study, bacterial ghosts (BGs) were generated from Weissella koreensis LKS42 (WKorGs) and Pediococcus pentosacues KA94 (PPGs) by chemically inducing lysis using substances such as hydrochloric acid (HCl), sulfuric acid (H2SO4), nitric acid (HNO3), acetic acid (CH3COOH), sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium carbonate (Na2CO3), n-butanol, and C6H8O7. HCl-induced WKorGs and PPGs exhibited complete removal of DNA and displayed transverse membrane dissolution tunnel structures under scanning electron microscopy (SEM). Cell viability assays showed high viability of RAW 264.7 cells exposed to HCl-induced WKorGs and PPGs. Additionally, treatment with HCl-induced WKorGs and PPGs elevated mRNA levels of pro-inflammatory cytokines (IL-1ß, IL-6, TNF-α, iNOS) and the anti-inflammatory cytokine IL-10 in RAW 264.7 cells. These findings suggest that HCl-induced WKorGs and PPGs have the potential to be used as inactivated bacterial immunostimulants, highlighting their promising applications in immunization and immunotherapy.

Effective Model Update for Adaptive Classification of Text Streams in a Distributed Learning Environment.

In this study, we propose dynamic model update methods for the adaptive classification model of text streams in a distributed learning environment. In particular, we present two model update strategies: (1) the entire model update and (2) the partial model update. The former aims to maximize the model accuracy by periodically rebuilding the model based on the accumulated datasets including recent datasets. Its learning time incrementally increases as the datasets increase, but we alleviate the learning overhead by the distributed learning of the model. The latter fine-tunes the model only with a limited number of recent datasets, noting that the data streams are dependent on a recent event. Therefore, it accelerates the learning speed while maintaining a certain level of accuracy. To verify the proposed update strategies, we extensively apply them to not only fully trainable language models based on CNN, RNN, and Bi-LSTM, but also a pre-trained embedding model based on BERT. Through extensive experiments using two real tweet streaming datasets, we show that the entire model update improves the classification accuracy of the pre-trained offline model; the partial model update also improves it, which shows comparable accuracy with the entire model update, while significantly increasing the learning speed. We also validate the scalability of the proposed distributed learning architecture by showing that the model learning and inference time decrease as the number of worker nodes increases.

Response of vesicle shapes to dense inner active matter.

We consider experimentally the Takatori-Sahu model of vesicle shape fluctuations induced by enclosed active matter, a model till present tested only in the absence of collective motion because few enclosed bacteria were used to generate the desired active motion (S. C. Takatori and A. Sahu, Phys. Rev. Lett., 2020, 124, 158102). Using deformable giant unilamellar vesicles (GUVs) and phase contrast microscopy, we extract the mode-dependence of GUV shape fluctuations when hundreds of E. coli bacteria are contained within each GUV. In the microscope focal plane, patterns of collective bacteria flow include vortex flow, dipolar flow, and chaotic motion, all of which influence the GUV shapes. The Takatori-Sahu model generalizes well to this situation if one considers the moving element to be the experimentally-determined size of the collecively-moving flock.

A bacteria-based carbon sequestration and waste recycling system.

Achieving carbon neutrality requires a variety of technological approaches. In the present study, we confirmed the applicability of a carbon cycle system in several industrial fields using sulphur-oxidising bacteria. This system produces a nitrogen fertiliser, which decreases carbon emissions by recycling H2S and NH3 pollutants discharged into the atmosphere or wastewater. It should be considered in industrial fields as a carbon reduction strategy.

Hydrochloric acid-treated Bacillus subtilis ghosts induce IL-1 beta, IL-6, and TNF-alpha in murine macrophage.

Background: Bacterial ghosts (BGs) are empty cell envelopes commonly generated using Gram-negative bacteria; they represent a potential platform for efficient adjuvant and vaccine delivery systems. However, the efficient production of BGs from bacteria in a short period of time is challenging. Objective: The purpose of this study was to investigate the possibility of producing BGs in the Gram-positive Bacillus subtilis using various chemicals, and the potential application of BGs as a novel immunomodulatory agent. Results: In this study, Bacillus subtilis ghosts (BSGs) were generated, for the first time to the best of our knowledge, using the minimum inhibitory concentration (MIC) of hydrochloric acid (HCl; 6.25 mg/mL), sulfuric acid (H2SO4; 3.125 mg/mL), and nitric acid (HNO3; 6.25 mg/mL). Among the BSGs generated using these chemicals, HCl-induced BSGs were completely DNA-free as confirmed by real-time polymerase chain reaction. Scanning electron microscopy showed the formation of transmembrane lysis tunnel structures in HCl-induced BSGs. Murine macrophages exposed to the HCl-induced BSGs at a concentration of 1 × 105 CFU/mL showed a cell viability of 97.8%. Additionally, HCl-induced BSGs upregulated the expression of pro-inflammatory cytokines including interleukin (IL)-1ß, tumor necrosis factor alpha, and IL-6. Furthermore, we found differences in the protein expression profiles between intact live bacteria and BSGs using two-dimensional electrophoresis coupled with peptide mass fingerprinting/matrix-assisted laser desorption/ionization-time of flight mass spectrometry analysis. Conclusion: These data suggest that the HCl-induced BSGs may be potentially safe and effective candidates for inactivated bacterial vaccines and/or immunostimulants. Supplementary Information: The online version contains supplementary material available at 10.1007/s13273-022-00221-5.

A comparative experimental study of distributed storage engines for big spatial data processing using GeoSpark.

With increasing numbers of GPS-equipped mobile devices, we are witnessing a deluge of spatial information that needs to be effectively and efficiently managed. Even though there are several distributed spatial data processing systems such as GeoSpark (Apache Sedona), the effects of underlying storage engines have not been well studied for spatial data processing. In this paper, we evaluate the performance of various distributed storage engines for processing large-scale spatial data using GeoSpark, a state-of-the-art distributed spatial data processing system running on top of Apache Spark. For our performance evaluation, we choose three distributed storage engines having different characteristics: (1) HDFS, (2) MongoDB, and (3) Amazon S3. To conduct our experimental study on a real cloud computing environment, we utilize Amazon EMR instances (up to 6 instances) for distributed spatial data processing. For the evaluation of big spatial data processing, we generate data sets considering four kinds of various data distributions and various data sizes up to one billion point records (38.5GB raw size). Through the extensive experiments, we measure the processing time of storage engines with the following variations: (1) sharding strategies in MongoDB, (2) caching effects, (3) data distributions, (4) data set sizes, (5) the number of running executors and storage nodes, and (6) the selectivity of queries. The major points observed from the experiments are summarized as follows. (1) The overall performance of MongoDB-based GeoSpark is degraded compared to HDFS- and S3-based GeoSpark in our experimental settings. (2) The performance of MongoDB-based GeoSpark is relatively improved in large-scale data sets compared to the others. (3) HDFS- and S3-based GeoSpark are more scalable to running executors and storage nodes compared to MongoDB-based GeoSpark. (4) The sharding strategy based on the spatial proximity significantly improves the performance of MongoDB-based GeoSpark. (5) S3- and HDFS-based GeoSpark show similar performances in all the environmental settings. (6) Caching in distributed environments improves the overall performance of spatial data processing. These results can be usefully utilized in decision-making of choosing the most adequate storage engine for big spatial data processing in a target distributed environment.

Shielding effect of radiation dose reduction fiber during the use of C-arm fluoroscopy: a phantom study.

This study evaluated the shielding effect of a newly developed dose-reduction fiber (DRF) made from barium sulfate, in terms of radiation doses delivered to patients' radiosensitive organs and operator during C-arm fluoroscopy and its impact on the quality of images. A C-arm fluoroscopy unit was placed beside a whole-body phantom. Radiophotoluminescent glass dosimeters were attached to the back and front of the whole-body phantom at 20 cm intervals. Radiation doses were measured without DRF and with it applied to the back (position 1), front (position 2) or both sides (position 3) of the phantom. To investigate the impact of DRF on the quality of fluoroscopic images, step-wedge and modulation transfer function phantoms were used. The absorbed radiation doses to the back of the phantom significantly decreased by 25.3-88.8% after applying DRF to positions 1 and 3. The absorbed radiation doses to the front of the phantom significantly decreased by 55.3-93.6% after applying DRF to positions 2 and 3. The contrast resolution values for each adjacent step area fell in the range 0.0119-0.0209, 0.0128-0.0271, 0.0135-0.0339 and 0.0152-0.0339 without and with DRF applied to positions 1, 2 and 3, respectively. The investigated DRF effectively reduces absorbed radiation doses to patients and operators without decreasing the quality of C-arm fluoroscopic images. Therefore, routine clinical use of the DRF is recommended during the use of C-arm fluoroscopy.

Apparatus to Measure Subnanometer Fluctuation of Giant Unilamellar Vesicle Membranes.

The design, calibration, and performance of an apparatus are described to study the nanometer-scale thermal or driven fluctuations of free-standing vesicle membranes using a design resembling the position detection system of optical tweezers except for the laser power lower by orders of magnitude to avoid trapping. Over four decades of frequency, 1-10,000 Hz, it reports membrane fluctuation amplitudes 0.01-100 nm by measuring scattering of a laser beam as it passes membranes (∼1 µm cross-section) suspended in the aqueous medium. The low-power laser beam, <100 µW, is sharply focused on the edge of a giant unilamellar vesicle, and fluctuations of position are measured using a position-sensitive photodetector. The central result of this approach is the capability to reach small fluctuations otherwise inaccessible using other techniques. The typical obtained data are fit to the standard Helfrich mechanical model. The applications and limitations of the device are discussed, as well as other potential uses to which the apparatus may be applied by rational extension of the approach presented.

Wavelength discrimination (WLD) TOF-PET detector with DOI information.

Depth-of-interaction (DOI) encoding can contribute to improving spatial resolution uniformity and sensitivity in positron-emission-tomography (PET) scanners. In addition, time-of-flight (TOF) PET scanners with DOI encoding have received considerable interest because of their potential for improving the spatial resolution, sensitivity, and image quality of the overall system. In this study, a new DOI detector configuration utilizing scintillators' emission wavelength is proposed, and experimental results on the energy, timing, and DOI performance of the detector are provided. The DOI information from the proposed phoswich-type detector can be acquired at the detector level without complex signal processing by utilizing a single optical filter with customized optical properties. For this, we used either a short pass filter (SPF) or a long pass filter (LPF) that allows light photons of a specific wavelength to pass. The two-layered phoswich detector was configured with two scintillators with different photon-emission spectra. In this study, we used Ce:GAGG (3 mm × 3 mm × 10 mm) and LYSO:Ce (3 mm × 3 mm × 10 mm) as the top and bottom layer scintillators, respectively. A digital silicon photomultiplier (dSiPM) was used as the photosensor and for data acquisition. The phoswich detector was placed in the center of two dSiPM pixels, where one of the dSiPM pixels was covered with the optical filter, and the light guide was placed on the other pixel. The detector was tested for energy, timing, and DOI encoding performance. When an SPF was used, the energy resolutions of 16.2% and 11.8% were achieved for the Ce:GAGG (top layer) and LYSO:Ce (bottom layer) respectively without correcting for saturation effect. With a small (3 mm × 3 mm × 5 mm) LYSO crystal as the reference detector, CRTs (coincidence-resolving times) of 338 ps and 244 ps were recorded for the top and bottom layers respectively. The detector configuration also provides an excellent DOI-separation figure-of-merit (FoM) value of 1.9. In the case of LPF, the energy resolutions of 12.0% and 12.9% were achieved for the Ce:GAGG (top layer) and LYSO:Ce (bottom layer), respectively. CRTs (coincidence resolving times) of 314 ps and 263 ps were recorded for the top and bottom layers, respectively. The DOI-separation FoM value of 1.5 was achieved in this setup. Results show that the proposed method can provide excellent discrete DOI positioning accuracy without compromising the timing performance of the detector.

A hybrid and scalable error correction algorithm for indel and substitution errors of long reads.

BACKGROUND: Long-read sequencing has shown the promises to overcome the short length limitations of second-generation sequencing by providing more complete assembly. However, the computation of the long sequencing reads is challenged by their higher error rates (e.g., 13% vs. 1%) and higher cost ($0.3 vs. $0.03 per Mbp) compared to the short reads. METHODS: In this paper, we present a new hybrid error correction tool, called ParLECH (Parallel Long-read Error Correction using Hybrid methodology). The error correction algorithm of ParLECH is distributed in nature and efficiently utilizes the k-mer coverage information of high throughput Illumina short-read sequences to rectify the PacBio long-read sequences.ParLECH first constructs a de Bruijn graph from the short reads, and then replaces the indel error regions of the long reads with their corresponding widest path (or maximum min-coverage path) in the short read-based de Bruijn graph. ParLECH then utilizes the k-mer coverage information of the short reads to divide each long read into a sequence of low and high coverage regions, followed by a majority voting to rectify each substituted error base. RESULTS: ParLECH outperforms latest state-of-the-art hybrid error correction methods on real PacBio datasets. Our experimental evaluation results demonstrate that ParLECH can correct large-scale real-world datasets in an accurate and scalable manner. ParLECH can correct the indel errors of human genome PacBio long reads (312 GB) with Illumina short reads (452 GB) in less than 29 h using 128 compute nodes. ParLECH can align more than 92% bases of an E. coli PacBio dataset with the reference genome, proving its accuracy. CONCLUSION: ParLECH can scale to over terabytes of sequencing data using hundreds of computing nodes. The proposed hybrid error correction methodology is novel and rectifies both indel and substitution errors present in the original long reads or newly introduced by the short reads.

Strategies to improve the photocatalytic activity of TiO2: 3D nanostructuring and heterostructuring with graphitic carbon nanomaterials.

TiO2-based photocatalysis has been considered to be one of the most promising avenues for environmental remediation including water purification. However, several technical issues such as the limited surface area of bulk TiO2, the large band gap of TiO2, and rapid charge recombination still limit the practical application of TiO2 photocatalysts. Therefore, here we focus on two structural design strategies: (i) monolithic three-dimensional (3D) nanostructuring, and (ii) heterostructuring with graphitic carbon nanomaterials. A monolithic 3D nanostructure enables maximal surface area in a given volume and efficient reuse of the photocatalyst without recollection. Heterostructuring with carbon nanomaterials helps achieve maximal utilization of the solar spectrum and charge separation and provides efficient TiO2 photocatalysts. In this review, recent progress on TiO2 photocatalysts toward the abovementioned strategies will be summarized. Further discussion and direction will provide insights into the rational design of highly efficient TiO2 photocatalysts, and help develop advanced photocatalyst models.

Effect of metallic tools on scattered radiation dose during the use of C-arm fluoroscopy in orthopaedic surgery.

This study investigated the effect of metallic tools on the scattered radiation dose delivered to surgeons' radiosensitive organs while simulating hip surgery using C-arm fluoroscopy. Two phantoms, a pelvis and a Rando phantom, were used to simulate a patient and a surgeon in this study. Photoluminescence dosimeters were inserted into the Rando phantom in the positions of the eye, thyroid and gonad. A drill was positioned above the hip of the pelvis phantom or beside the pelvis phantom of the same height. For each drill location, the scattered radiation dose was measured when the angle to the operator phantom was 45°; this was repeated when the angle was 90°. The scattered radiation doses to the eye, thyroid and gonad when the drill was placed beside the pelvis phantom with 90° angulation to the operator phantom were significantly lower than the reference values and those when the drill was placed beside the pelvis phantom at a 45° angulation to the operator phantom. The scattered radiation doses to the eye and thyroid when the drill was placed above the hip were significantly lower than the references values. Of the four different scenarios, the scattered radiation doses to the eye, thyroid and gonad were lowest when the drill was placed beside the pelvis phantom with 90° angulation. This study showed that the scattered radiation doses to radiosensitive organs were affected by the location and angle of the metallic tools in relation to the operator. Therefore, orthopedic surgeons should consider the effect of metallic tools on the scattered radiation dose during intraoperative use of C-arm fluoroscopy.

Dextran adsorption onto red blood cells revisited: single cell quantification by laser tweezers combined with microfluidics.

The aggregation of red blood cells (RBC) is of importance for hemorheology, while its mechanism remains debatable. The key question is the role of the adsorption of macromolecules on RBC membranes, which may act as "bridges" between cells. It is especially important that dextran is considered to induce "bridge"-less aggregation due to the depletion forces. We revisit the dextran-RBC interaction on the single cell level using the laser tweezers combined with microfluidic technology and fluorescence microscopy. An immediate sorption of ~104 molecules of 70 kDa dextran per cell was observed. During the incubation of RBC with dextran, a gradual tenfold increase of adsorption was found, accompanied by a moderate change in the RBC deformability. The obtained data demonstrate that dextran sorption and incubation-induced changes of the membrane properties must be considered when studying RBC aggregation in vitro.

An iterative sinogram gap-filling method with object- and scanner-dedicated discrete cosine transform (DCT)-domain filters for high resolution PET scanners.

PURPOSE: We aimed to develop a gap-filling algorithm, in particular the filter mask design method of the algorithm, which optimizes the filter to the imaging object by an adaptive and iterative process, rather than by manual means. METHODS: Two numerical phantoms (Shepp-Logan and Jaszczak) were used for sinogram generation. The algorithm works iteratively, not only on the gap-filling iteration but also on the mask generation, to identify the object-dedicated low frequency area in the DCT-domain that is to be preserved. We redefine the low frequency preserving region of the filter mask at every gap-filling iteration, and the region verges on the property of the original image in the DCT domain. RESULTS: The previous DCT2 mask for each phantom case had been manually well optimized, and the results show little difference from the reference image and sinogram. We observed little or no difference between the results of the manually optimized DCT2 algorithm and those of the proposed algorithm. CONCLUSIONS: The proposed algorithm works well for various types of scanning object and shows results that compare to those of the manually optimized DCT2 algorithm without perfect or full information of the imaging object.

A Salmonella typhimurium ghost vaccine induces cytokine expression in vitro and immune responses in vivo and protects rats against homologous and heterologous challenges.

Salmonella enteritidis and Salmonella typhimurium are important food-borne bacterial pathogens, which are responsible for diarrhea and gastroenteritis in humans and animals. In this study, S. typhimurium bacterial ghost (STG) was generated based on minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH). Experimental studies performed using in vitro and in vivo experimental model systems to characterize effects of STG as a vaccine candidate. When compared with murine macrophages (RAW 264.7) exposed to PBS buffer (98.1%), the macrophages exposed to formalin-killed inactivated cells (FKC), live wild-type bacterial cells and NaOH-induced STG at 1 × 108 CFU/mL showed 85.6%, 66.5% and 84.6% cell viability, respectively. It suggests that STG significantly reduces the cytotoxic effect of wild-type bacterial cells. Furthermore, STG is an excellent inducer for mRNAs of pro-inflammatory cytokine (TNF-α, IL-1ß) and factor (iNOS), anti-inflammatory cytokine (IL-10) and dual activities (IL-6) in the stimulated macrophage cells. In vivo, STG vaccine induced humoral and cellular immune responses and protection against homologous and heterologous challenges in rats. Furthermore, the immunogenicity and protective efficacy of STG vaccine were compared with those of FKC and non-vaccinated PBS control groups. The vaccinated rats from STG group exhibited higher levels of serum IgG antibody responses, serum bactericidal antibodies, and CD4+ and CD8+ T-cell populations than those of the FKC and PBS control groups. Most importantly, after challenge with homologous and heterologous strains, the bacterial loads in the STG group were markedly lower than the FKC and PBS control groups. In conclusion, these findings suggest that the STG vaccine induces protective immunity against homologous and heterologous challenges.

Assessment of the "cross-bridge"-induced interaction of red blood cells by optical trapping combined with microfluidics.

Red blood cell (RBC) aggregation is an intrinsic property of the blood that has a direct effect on the blood viscosity and circulation. Nevertheless, the mechanism behind the RBC aggregation has not been confirmed and is still under investigation with two major hypotheses, known as "depletion layer" and "cross-bridging." We aim to ultimately understand the mechanism of the RBC aggregation and clarify both models. To measure the cell interaction in vitro in different suspensions (including plasma, isotonic solution of fibrinogen, isotonic solution of fibrinogen with albumin, and phosphate buffer saline) while moving the aggregate from one solution to another, an approach combining optical trapping and microfluidics has been applied. The study reveals evidence that RBC aggregation in plasma is at least partly due to the cross-bridging mechanism. The cell interaction strength measured in the final solution was found to be significantly changed depending on the initial solution where the aggregate was formed.

Effect of shear-induced platelet activation on red blood cell aggregation.

Mechanical shear stress is one of the important factors for platelet activation. Although shear stress has been frequently utilized in many applications of diagnostic bio-equipment, there has been little consideration as to whether shear stress induces platelet activation and consequently alters hemorheological characteristics. Therefore, we investigated the effect of shear-induced platelet activation on red blood cell (RBC) aggregation. The hypothesis of the present research is as follows: Platelets activated by high shear stress secrete substances, which can affect hemorheological characteristics to promote RBC aggregation. In our study, an increase in RBC aggregation indices (critical shear stress (CSS) and aggregation index (AI)) by shear-induced platelet activation was observed. Significantly, an increase of 19% in CSS was observed. However, deformability remained unchanged. These phenomena could be a result of the increased cellular adhesion force on RBC membranes due to secreted substances from activated platelets. Therefore, since high shear application results in the unexpected effect on RBC aggregation, conditions for shear application in diagnostic bio-equipment are to be carefully determined.

Simulation study of a novel target oriented SPECT design using a variable pinhole collimator.

PURPOSE: In the past decade, demands for organ specific (target oriented) single-photon emission computed tomography (SPECT) is increasing, and several groups have conducted studies on developing clinical dedicated SPECT with pinhole collimator to improve the spatial resolution. However, acceptance angle of the collimator cannot be adjusted to fit the different ROIs of target objects because the shape of pinhole could not be changed, and the magnifying factor cannot be maximized as the collimator-to-detector distance is fixed. Furthermore, those dedicated pinhole SPECTs are typically made for a single purpose and therefore possess a drawback in that it cannot be utilized for any other purpose. In this study, we propose a novel SPECT system using variable pinhole collimator (VP SPECT) whose parameters are flexible. METHODS: The proposed variable pinhole collimator is modeled on conventional pinhole by piling several tungsten layers of different apertures. Depending on the combination of the holes in each layer, a variety of hole diameters and acceptance angles of the pinhole can be made. In addition, VP SPECT system allows attaching the collimator to the object as close as possible to maximize the sensitivity and adjust the distance of the pinhole from the scintillation detector to optimize the system resolution for each rotation angle, automatically. For quantitative measurement, we compared the sensitivity and spatial resolution of VP SPECT with those of conventional pinhole SPECT. To determine the possibility of the clinical and preclinical use of proposed system, a digital mouse whole-body (MOBY) phantom is used for simulating the live mouse model. RESULTS: The result of simulation using ultra-micro hot spot phantom shows that the sensitivity of the proposed VP SPECT system is about 297% of that of the conventional system. While hot rods of diameter 0.6 mm can be distinguished in the image with the proposed VP SPECT system, 1.2-mm hot rods are barely discernible in the conventional pinhole SPECT image. According to the result of MOBY phantom simulation, heart walls separated by 3 mm were not distinguished in conventional pinhole SPECT images, but were clearly discerned in VP SPECT images. CONCLUSIONS: In this study, we designed a novel pinhole collimator for SPECT and presented preliminary results of target oriented imaging with a simulation study. Currently, we are pursuing strategies to realize the proposed system, with the goal to apply the technology into a high-sensitivity and high-resolution preclinical SPECT. Should VP SPECT be applied to the clinical setting, we anticipate a high-sensitivity, high-resolution system for applications such as heart dedicated SPECT or related fields.

Characterization of Chemically-Induced Bacterial Ghosts (BGs) Using Sodium Hydroxide-Induced Vibrio parahaemolyticus Ghosts (VPGs).

Acellular bacterial ghosts (BGs) are empty non-living bacterial cell envelopes, commonly generated by controlled expression of the cloned lysis gene E of bacteriophage PhiX174. In this study, Vibrio parahaemolyticus ghosts (VPGs) were generated by chemically-induced lysis and the method is based on minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH), acetic acid, boric acid, citric acid, maleic acid, hydrochloric acid, and sulfuric acid. The MIC values of the respective chemicals were 3.125, 6.25, <50.0, 25.0, 6.25, 1.56, and 0.781 mg/mL. Except for boric acid, the lysis efficiency reached more than 99.99% at 5 min after treatment of all chemicals. Among those chemicals, NaOH-induced VPGs appeared completely DNA-free, which was confirmed by quantitative real-time PCR. Besides, lipopolysaccharides (LPS) extracted from the NaOH-induced VPGs showed no distinctive band on SDS-PAGE gel after silver staining. On the other hand, LPS extracted from wild-type bacterial cells, as well as the organic acids-induced VPGs showed triple major bands and LPS extracted from the inorganic acids-induced VPGs showed double bands. It suggests that some surface structures in LPS of the NaOH-induced VPGs may be lost, weakened, or modified by the MIC of NaOH. Nevertheless, Limulus amoebocyte lysate assay revealed that there is no significant difference in endotoxic activity between the NaOH-induced VPGs and wild-type bacterial cells. Macrophages exposed to the NaOH-induced VPGs at 0.5 × 106 CFU/mL showed cell viability of 97.9%, however, the MIC of NaOH did not reduce the cytotoxic effect of wild-type bacterial cells. Like Escherichia coli LPS, the NaOH-induced VPGs are an excellent activator of pro-inflammatory cytokines (IL-1ß and iNOS), anti-inflammatory cytokine (IL-10), and dual activities (IL-6) in the stimulated macrophage cells. On the other hand, the induction of TNF-α mRNA was remarkable in the macrophages exposed with wild-type cells. Scanning electron microscopy showed the formation of trans-membrane lysis tunnel structures in the NaOH-induced VPGs. SDS-PAGE and agarose gel electrophoresis also confirmed that cytoplasmic proteins and genomic DNA released from the VPGs to culture medium through the lysis tunnel structures. Taken together, all these data indicate that the NaOH-induced VPGs show the potency of a safe, economical, and effective inactivated bacterial vaccine candidate.

Performances of a protector against scattered radiation during intraoperative use of a C-arm fluoroscope.

The scattered radiation protector for mobile x-ray systems, Creative Valuable Protector-2, has been recently developed. However, there have been no studies investigating the effects of this device. We aim to investigate the effects of the scattered radiation protector on the equivalent doses from scattered radiation delivered to radiosensitive organs while simulating spine surgery using a C-arm fluoroscope. Chest and rando phantoms were used to simulate a patient and a surgeon in this study. The equivalent dose from scattered radiation to radiosensitive organs was measured in four different situations according to the use of the scattered radiation protector and the C-arm configuration. To compare the quality of the images with and without the scattered radiation protector, an acryl step phantom with five steps was used, and the contrast resolution of each step was calculated. The equivalent dose from the scattered radiation to the surgeon's eye, thyroid, and gonad decreased significantly by using the scattered radiation protector for both the Posteroanterior (PA) (p < 0.001) and Anteroposterior (AP) (p < 0.001) C-arm configurations. The installation of the scattered radiation protector also reduced the direct radiation dose to the chest phantom. A scattered map showed that scattered radiation doses decreased by approximately 50% for the PA configuration and 75% for the AP configuration by using the scattered radiation protector. Before and after installation of the scattered radiation protector, the contrast resolution of each adjacent step area was 0.025-0.404 and 0.216-0.421. The scattered radiation protector was effective in reducing not only the equivalent dose from scattered radiation to the surgeon's radiosensitive organs, but also the direct radiation dose to the patient. This was all achieved without decreasing the quality of the C-arm fluoroscopic images.