Development of a Novel ATP Bioluminescence Assay Based on Engineered Probiotic Saccharomyces boulardii Expressing Firefly Luciferase.

Quantitative analysis of adenosine triphosphate (ATP) has been widely used as a diagnostic tool in the food and medical industries. Particularly, the pathogenesis of a few diseases including inflammatory bowel disease (IBD) is closely related to high ATP concentrations. A bioluminescent D-luciferin/luciferase system, which includes a luciferase (FLuc) from the firefly Photinus pyralis as a key component, is the most commonly used method for the detection and quantification of ATP. Here, instead of isolating FLuc produced in recombinant Escherichia coli, we aimed to develop a whole-cell biocatalyst system that does not require extraction and purification of FLuc. To this end, the gene coding for FLuc was introduced into the genome of probiotic Saccharomyces boulardii using the CRISPR/Cas9-based genome editing system. The linear relationship (r2 = 0.9561) between ATP levels and bioluminescence generated from the engineered S. boulardii expressing FLuc was observed in vitro. To explore the feasibility of using the engineered S. boulardii expressing FLuc as a whole-cell biosensor to detect inflammation biomarker (i.e., ATP) in the gut, a colitis mouse model was established using dextran sodium sulfate as a colitogenic compound. Our findings demonstrated that the whole-cell biosensor can detect elevated ATP levels during gut inflammation in mice. Therefore, the simple and powerful method developed herein could be applied for non-invasive IBD diagnosis.

Erratum to: Methylobacterium terrae sp. nov., a radiation-resistant bacterium isolated from gamma ray-irradiated soil.

In the article by Kim et al. published in Journal of Microbiology 2019; 57, 959-966, The NBRC accession number NBRC 112879T on 33th line of 2nd paragraph in the section of 'Description of Methylobacterium terrae sp. nov.' on page 964 should be corrected in NBRC 112873T.The sentence in abstract should have read: The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene and genome sequences of the type strain 17Sr1-28T (= KCTC 52904T = NBRC 112873T) are KY939566 and CP029553, respectively.We apologize for any inconvenience that this may have caused.

Sphingomonas edaphi sp. nov., a novel species isolated from beach soil in the Republic of Korea.

A yellow-coloured, Gram-negative, motile, strictly aerobic bacterial strain, designated strain DAC4T, was isolated from a soil sample collected at Ahnmok Beach (Busan, Republic of Korea). The cells of strain DAC4T were rod-shaped and the colonies that formed were round and convex. The results of phylogenetic analysis based on the 16S rRNA gene sequence of strain DAC4T revealed that the bacterium belongs to the genus Sphingomonas, family Sphingomonadaceae, and that it was most closely related to Sphingomonas jaspsi DSM 18422T (98.01 %), Sphingomonas rhizophila KACC 19189T (97.76 %), Sphingomonas mesophila KCTC 62179T(97.30 %), Sphingomonas sedimincola KCTC 12629T (97.16 %) and Sphingomonas oryziterrae KCTC 22476T (97.05 %). The major respiratory quinone was Q-10, and the major cellular fatty acids were summed feature 8 (C18  :  1ω7c) and summed feature 3 (C16  :  1ω7c/C16  :  1ω6c). The whole genome DNA G+C content of strain DAC4T was 62.16 mol%. Phosphatidylethanolamine, diphosphatidylglycerol, sphingoglycolipids, phosphatidylglycerol, phosphatidylcholine, four undefined glycolipids and an undefined lipid were detected in strain DAC4T, and the strain had sym-homospermidine as a major polyamine. The in silico DNA-DNA hybridization and average nucleotide identity values between strain DAC4T and the closely related taxa S. jaspsi and S. mesophila were 75.5/23.5 % and 73.5 /18.5%, respectively. The fluorimetric DNA-DNA hybridization results showed that strain DAC4T and S. rhizophila, S. sediminicola and S. oryziterrae have 37.1, 35.2 and 32.2 % DNA similarity, respectively. Based on phylogenetic, phenotypic and chemotaxonomic distinctiveness, strain DAC4T (=KCTC 62107T=JCM 32377T) is classified as a novel species of the genus Sphingomonas, for which the name Sphingomonas edaphi sp. nov. is proposed.

Kaposi's sarcoma-associated herpesvirus viral protein kinase phosphorylates extracellular signal-regulated kinase and activates MAPK/ERK signaling pathway.

Open reading frame 36 (ORF36) of Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a serine/threonine-type viral protein kinase (vPK). Previous studies have examined the functions of KSHV vPK; however, its role in the activation of extracellular signal-regulated kinase (ERK1/2) has not yet been described to date. Using HEK 293 cell lines, we performed a human phospho-kinase array analysis to screen for MAPK signaling pathways kinases that are activated by KSHV vPK. In addition, we investigated the regulator protein phosphorylation of up/downstream ERK1/2 pathway; nuclear translocation of phosphorylated ERK1/2; and regulation of transcription factor, inflammatory cytokine, and pro-/anti-apoptotic factor by KSHV vPK transfection. Here, we demonstrated that KSHV vPK activates ERK1/2 signaling pathway and plays an important role in the activation of MAPK/ERK signaling pathway.

Methylobacterium terrae sp. nov., a radiation-resistant bacterium isolated from gamma ray-irradiated soil.

A Gram-stain-negative, asporogenous, aerobic rods, motile by means of a single polar flagellum, catalase- and oxidase-positive, methylotrophic bacterium, designated 17Sr1-28T, was isolated from gamma ray-irradiated soil. The 16S rRNA gene sequence analysis showed that strain 17Sr1-28T was phylogenetically related to Methylobacterium currus PR1016AT (96.8%), Methylobacterium platani PMB02T (96.2%), Methylobacterium aquaticum DSM 16371T (96.3%), Methylobacterium tarhaniae N4211T (96.4%), Methylobacterium frigidaeris IER25-16T (95.8%), and Methylobacterium organophilum JCM 2833T (92.7%). The G+C content calculated based on genome sequence was 71.6%. The average nucleotide identity and in silico DNA-DNA hybridization values between strain 17Sr1- 28T and M. currus, M. platani, M. aquaticum, M. tarhaniae, M. frigidaeris, and M. organophilum were 77.7-90.4% and 22-39.6%, respectively. The major fatty acids of strain 17Sr1-28T were summed feature 8 (C18:1ω7c and/or C18:1ω6c), and summed feature 3 (C16:1ω7c and/or C16:1ω6c). The predominant quinone was ubiquinone 10 and the major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol. On the basis of the data from phenotypic tests and genotypic differences between strain 17Sr1-28T and its close phylogenetic relatives, strain 17Sr1-28T represents a new species belonging to the genus Methylobacterium, for which the name Methylobacterium terrae sp. nov. (= KCTC 52904T = NBRC 112873T) is proposed.

Ilyomonas limi gen. nov., sp. nov., a new member of the family Chitinophagaceae isolated from mud.

A Gram-strain negative, aerobic, catalase and oxidase positive, non-motile, short rod-shaped bacterium, designated 17mud1-8T, was isolated from mud collected from Nowon-gu, Seoul, South Korea. The strain was found to be able to grow at 10-40 °C (optimum 28-30 °C), pH 5.0-8.0 (optimum 7.0), and in the absence of NaCl. The nearly full-length 16S rRNA gene of strain 17mud1-8T exhibits sequence similarity of 94.1% with that of Panacibacter ginsenosidivorans Gsoil 1550T, followed by 93.6% sequence similarity with Parafilimonas terrae DSM 28286T. Phylogenetic analysis indicated that strain 17mud1-8T belongs to the family Chitinophagaceae, sharing approximately 94.1-91.9% sequence similarity with members of closely related genera. The respiratory quinone was identified as MK-7. The predominant fatty acids were found to consist of iso-C15:0, iso-C17:1ω5c and iso-C15:1 G. The polar lipids were identified as phosphatidylethanolamine, an unidentified aminophospholipid, an unidentified glycolipid, ten unidentified aminolipids and seven unidentified lipids. The draft genome of 17mud1-8T has G+C content of 40.9 mol% and a 5.8 Mb chromosome. On the basis of the phenotypic and genotypic properties, and phylogenetic inference, strain 17mud1-8T was found to represent a novel genus in the family Chitinophagaceae, for which the name Ilyomonas limi gen. nov., sp. nov. is proposed, with the type strain 17mud1-8T(=KCTC 52874T = NBRC 112826T).

Pontibacter oryzae sp. nov., a carotenoid-producing species isolated from a rice paddy field.

A taxonomic study using a polyphasic approach was performed on a Gram-stain negative, red-pink, aerobic, non-motile, asporogenous, rod-shaped bacterium, designated strain KIRANT, isolated from soil collected from a rice paddy field. The 16S rRNA gene sequence analysis showed that strain KIRANT is phylogenetically related to Pontibacter actiniarum KMM 6156T, Pontibacter korlensis X14-1T, Pontibacter odishensis JC130T, Pontibacter litorisediminis YKTF-7T and Pontibacter aurantiacus NP1T (97.6, 97.5, 97.3, 97.3 and 96.7% sequence similarity, respectively). The major fatty acids of strain KIRANT were identified as iso-C15:0, iso-C15:0 3-OH and summed feature 4. The predominant menaquinone was identified as MK-7. The polar lipid profile was found to consist of phosphatidylethanolamine, four unidentified phospholipids, an unidentified glycolipid, an unidentified aminolipid and four unidentified lipids. The genome of strain KIRANT has a G + C content of 48.3 mol%. The in silico DNA-DNA hybridization and average nucleotide identity values between strain KIRANT and the closely related strains P. actiniarum KMM 6156T and P. korlensis X14-1T were 21.2%/21.8% and 76.4%/75.1%, respectively. On the basis of the data from phenotypic tests and genotypic differences between strain KIRANT and its close phylogenetic relatives, strain KIRANT is concluded to represent a new species belonging to the genus Pontibacter, for which the name Pontibacter oryzae sp. nov. is proposed. The type strain is KIRANT (= KACC 19815T = JCM 32880T).

Assessment of portal image resolution improvement using an external aluminum target and polystyrene electron filter.

BACKGROUND: In this study, an external 8 mm thick aluminum target was installed on the upper accessory tray mount of a medical linear accelerator head. The purpose of this study was to determine the effects of the external aluminum target beam (Al-target beam) on the portal image quality by analyzing the spatial and contrast resolutions. In addition, the image resolutions with the Al-target beams were compared with those of conventional 6 megavoltage (MV) images. METHODS: The optimized Al-target beam was calculated using Monte Carlo simulations. To validate the simulations, the percentage depth dose and lateral profiles were measured and compared with the modeled dose distributions. A PTW resolution phantom was used for imaging to assess the image resolution. The spatial resolution was quantified by determining the modulation transfer function. The contrast resolution was determined by a fine contrast difference between the 27 measurement areas. The spatial and contrast resolutions were compared with the those of conventional portal images. RESULTS: The measured and calculated percentage depth dose of the Al-target beam were consistent within 1.6%. The correspondence of measured and modelled profiles was evaluated by gamma analysis (3%, 3 mm) and all gamma values inside the field were less than one. The critical spatial frequencies (f50) of the images obtained with the Al-target beam and conventional imaging beam were 0.745 lp/mm and 0.451 lp/mm, respectively. The limiting spatial frequencies (f10) for the Al-target beam image and the conventional portal image were 2.39 lp/mm and 1.82 lp/mm, respectively. The Al-target beam resolved the smaller and lower contrast objects better than that of the MV photon beam. CONCLUSION: The Al-target beams generated by the simple target installation method provided better spatial and contrast resolutions than those of the conventional 6 MV imaging beam.

Percutaneous Endoscopic Lumbar Foraminoplasty for Lumbar Foraminal Stenosis of Elderly Patients with Unilateral Radiculopathy: Radiographic Changes in Magnetic Resonance Images.

BACKGROUND AND STUDY AIMS/OBJECTIVE: Clinical outcomes of percutaneous endoscopic lumbar foraminoplasty (PELF) performed for lumbar foraminal stenosis (LFS) have been reported in the literature. However, no study has reported the radiographic changes in magnetic resonance imaging (MRI) after surgery. This report presents the clinical outcome as well as radiographic changes in MRI after PELF for elderly patients with unilateral radiculopathy. We also describe detailed operative nuances to expand the foraminal space and mobilize the affected nerve root. MATERIAL AND METHODS: Between January 2015 and December 2016, 24 patients over 65 years of age underwent PELF to treat unilateral radiculopathy caused by LFS. The demographics, medical history, visual analog scale (VAS), and Oswestry Disability Index (ODI) were reviewed retrospectively. MRI radiographic data, cross-sectional area (CSA) of the affected foramen and nerve root, were evaluated before and after surgery. RESULTS: Preoperative radiculopathy improved in 23 of 24 patients (96%). The VAS score significantly improved from 7.89 ± 1.8 to 2.57 ± 2.5 (67.4%). The ODI also significantly improved from 33.15 ± 9.2 to 10.24 ± 6.7 (69.1%). Foraminal width increased ∼ 1.67 mm (21.4%), foraminal height increased 5.00 mm (36.9%), and CSA increased about 55.27 mm2 (60.6%) (p < 0.001). The CSA and the shape of the affected nerve section also expanded, but these were not statistically significant. CONCLUSION: We performed PELF in elderly patients with LFS. There were considerable radiographic changes in MRI, and the patients also showed clinically relevant improvement. Thus PELF resulted in sufficient decompression of the nerve root in LFS.

Investigation of the use of external aluminium targets for portal imaging in a medical accelerator using Geant4 Monte Carlo simulation.

OBJECTIVE: To install a low-Z target on the wedge tray mount of a medical linear accelerator to create a new image beam and to confirm image contrast enhancement. METHODS: Experimental low-energy photon beams were produced with the linac running in the 6 MeV electron mode and with a low-Z target installed on the wedge tray mount [denoted 6 MeV (low-Z target)]. Geant4 Monte Carlo simulation was performed to analyse the energy spectrum and image contrast of a 6 MeV (low-Z target) beam. This study modelled the 6 MeV (low-Z target) beam and the 6 MV (megavoltage) radiotherapy photon beam and verified model validity by measurement. In addition, a contrast phantom was modelled to quantitatively compare the image contrasts of the 6 MeV (low-Z target) beam and the 6 MV radiotherapy photon beam. A low-Z target was fabricated to generate low-energy photons (25-150 keV) from incident electrons, and a portal image of the Alderson RANDO phantom was acquired using a clinical linear accelerator for qualitative analysis. RESULTS: The measured and calculated percentage depth dose of the 6 MV photon and 6 MeV (Al) beams were consistent within 1.5 and 1.6%, respectively, and calculated lateral profiles of the 6 MV photon beam and the 6 MeV (Al) beam were consistent with the measured results within 1.5 and 1.9%, respectively. Although low-energy photons (25-150 keV) of the 6 MV photon beam were only 0.3%, the Be, C, and Al low-Z targets, but not the Ti target, generated 34.4 to 38.5% low-energy photons. In 5 to 20 cm water phantoms, contrast of the 6 MeV (Al) beam was approximately 1.16 times greater than that of the 6 MV beam. The contrasts of 6 MeV (Al) and 6 MV photon beams in the 20 cm water phantom were ~34% lower than those in the 5 cm water phantom. 6 MeV (Al)/CR (computed radiography) images of the human body phantom were more vivid and detailed than 6 MV/EPID (electronic portal imaging device) and 6 MeV (Al)/EPID images. CONCLUSION: The experimental beam with a low-Z target, which was simply installed on the wedge tray mount of the radiotherapy linear accelerator, generated significantly more low-energy photons than the 6 MV radiotherapy photon beam, and provided better quality portal images. Advances in knowledge: This study shows that, unlike the existing low-Z beam studies, a low-Z target can be installed outside the head of a linear accelerator to improve portal image quality.

Olfactory schwannoma-case report-.

Intracranial schwannomas preferentially arise from the vestibular branch of the eighth nerve, and rarely from the trigeminal nerve, facial nerve, and lower cranial nerves. Anterior cranial fossa schwannomas are extremely uncommon and few details about them have been reported. The patient was a 39-year-old woman whose chief complaints were anosmia and frontal headache for 2 years. The gadolinium (Gd)-enhanced magnetic resonance imaging (MRI) showed an extra-axial mass from ethmoid sinus to right frontal base region near the midline, with solid enhancement in lower portion and multicystic formation in upper portion. The tumor was totally resected via basal subfrontal approach. At operation, the tumor had cystic portion with marginal calcification and the anterior skull base was destructed by the tumor. The olfactory bulb was involved, and the tumor capsule did not contain neoplastic cells. The histopathological diagnosis was schwannoma. We report a rare case of anterior cranial fossa schwannoma with literature review.

Removal of intradural-extramedullary spinal cord tumors with unilateral limited laminectomy.

Total laminectomy for the removal of intradural-extramedullary spinal cord tumors has been used widely, but postoperative complications often develop, such as kyphosis, spinal instability, and persistent back pain. In this study, we evaluated seven patients with intradural-extramedullary spinal cord tumors with respect to the value of unilateral limited laminectomy. Our cases included six schwannomas, and one meningioma. The cervical region was involved in four cases, the thoracolumbar region in two cases, and the lumbar region in one case. The rationale for choosing a unilateral approach is to preserve musculoligamentous attachments and posterior bony elements as much as possible. The patients were mobilized on the third postoperative day and preoperative neurological symptoms were recovered within a few weeks. We did not observe any complication relating to unilateral limited laminectomy and at follow-up evaluation (at 3 and 12 months postoperatively), none of the patients showed spinal deformity or spinal instability. We think that the unilateral limited laminectomy is a safe and efficient technique for the treatment of intradural-extramedullary spinal cord tumors. We suggest that this technique is one of the best treatments for these tumors.

Predisposing factors related to shunt-dependent chronic hydrocephalus after aneurysmal subarachnoid hemorrhage.

OBJECTIVE: Hydrocephalus is a common sequelae of aneurysmal subarachnoid hemorrhage (SAH) and patients who develop hydrocephalus after SAH typically have a worse prognosis than those who do not. This study was designed to identify factors predictive of shunt-dependent chronic hydrocephalus among patients with aneurysmal SAH, and patients who require permanent cerebrospinal fluid diversion. METHODS: Seven-hundred-and-thirty-four patients with aneurysmal SAH who were treated surgically between 1990 and 2006 were retrospectively studied. Three stages of hydrocephalus have been categorized in this paper, i.e., acute (0-3 days after SAH), subacute (4-13 days after SAH), chronic (>/=14 days after SAH). Criteria indicating the occurrence of hydrocephalus were the presence of significantly enlarged temporal horns or ratio of frontal horn to maximal biparietal diameter more than 30% in computerized tomography. RESULTS: Overall, 66 of the 734 patients (8.9%) underwent shunting procedures for the treatment of chronic hydrocephalus. Statistically significant associations among the following factors and shunt-dependent chronic hydrocephalus were observed. (1) Increased age (p < 0.05), (2) poor Hunt and Hess grade at admission (p < 0.05), (3) intraventricular hemorrhage (p < 0.05), (4) Fisher grade III, IV at admission (p < 0.05), (5) radiological hydrocephalus at admission (p < 0.05), and (6) post surgery meningitis (p < 0.05) did affect development of chronic hydrocephalus. However the presence of intracerebral hemorrhage, multiple aneurysms, vasospasm, and gender did not influence on the development of shunt-dependent chronic hydrocephalus. In addition, the location of the ruptured aneurysms in posterior cerebral circulation did not correlate with the development of shunt-dependent chronic hydrocephalus. CONCLUSION: Hydrocephalus after aneurysmal SAH seems to have a multifactorial etiology. Understanding predisposing factors related to the shunt-dependent chronic hydrocephalus may help to guide neurosurgeons for better treatment outcomes.