Mutation of the N-terminal proline 9 of BLMA from Streptomyces verticillus abolishes the binding affinity for bleomycin

AbstractA gene, blmA, from bleomycin (Bm)-producing Streptomyces verticillus, encodes a Bm-binding protein, designated BLMA. The expression of BLMA conferred resistance to Bm in the Escherichia coli host, whereas a mutant protein, designated Pro-9/Leu, with the N-terminal proline 9 residue in BLMA replaced by leucine, did not. We created a fusion protein between the maltose-binding protein (MBP) and a mutant protein Pro-9/Leu/Leu with Met-94 in Pro-9/Leu replaced by leucine. Pro-9/Leu/Leu from the fusion protein, obtained by digestion with CNBr digestion, did not inhibit DNA-cleaving and antibacterial activities of Bm. Native-polyacrylamide gel electrophoresis (PAGE) and gel filtration column chromatographic analysis showed that the molecular size of Pro-9/Leu/Leu is roughly half of that of BLMA, suggesting that the mutant protein cannot form dimeric structure. Furthermore, Far-UV circular dichroism (CD) spectrum of Pro-9/Leu/Leu was quite different from that of BLMA and similar to the spectra obtained from unordered proteins [Venyaminov, S.Y. and Vassilenko, K.S. (1994) Anal. Biochem. 222, 176–184], suggesting that the secondary structure of Pro-9/Leu/Leu is disrupted. These results indicate that the mutation abolishes not only dimer formation but also the secondary structure of BLMA, which results in the loss of its function as a Bm-resistance determinant

造形表現に関する概念規定について : イメージ・アイデア・具象と抽象

Art is originally the projection which is reflected by fermenting and subliming the immanent value\u27s view of personality on the creative opportunity. In addition to this, the area of design requirs the functional quality based on the logic of lawful object. Finally it can be said that the design is the fact. It synthesizes the relation between the function and the sensuous and immanent beauty. The concept of \u27Image\u27, \u27Idea\u27 and \u27Concrete and Abstract\u27 is considered in this thesis. Judging from the designing point of view, they have the high frequency in use in the daily term. And this thesis aims at clearing up the concept which was vaguely used hitherto

A Raman Lidar with a Deep Ultraviolet Laser for Continuous Water Vapor Profiling in the Atmospheric Boundary Layer

A Raman lidar with a deep ultraviolet laser was constructed to continuously monitor water vapor distributions in the atmospheric boundary layer for twenty-four hours. We employ a laser at a wavelength of 266 nm and detects the light separated into an elastic backscatter signal and vibrational Raman signals of oxygen, nitrogen, and water vapor. The lidar was encased in a temperature-controlled and vibration-isolated compact container, resistant to a variety of environmental conditions. Water vapor profile observations were made for twelve months from November 24, 2017, to November 29, 2018. These observations were compared with collocated radiosonde measurements for daytime and nighttime conditions

Analysis of the Relationship between Hypertrophy of the Ligamentum Flavum and Lumbar Segmental Motion with Aging Process

Study DesignRetrospective cross-sectional study.PurposeTo investigate the relationship between ligamentum flavum (LF) hypertrophy and lumbar segmental motion.Overview of LiteratureThe pathogenesis of LF thickening is unclear and whether the thickening results from tissue hypertrophy or buckling remains controversial.Methods296 consecutive patients underwent assessment of the lumbar spine by radiographic and magnetic resonance imaging (MRI). Of these patients, 39 with normal L4–L5 disc height were selected to exclude LF buckling as one component of LF hypertrophy. The study group included 27 men and 12 women, with an average age of 61.2 years (range, 23–81 years). Disc degeneration and LF thickness were quantified on MRI. Lumbar segmental spine instability and presence of a vacuum phenomenon were identified on radiographic images.ResultsThe distribution of disc degeneration and LF thickness included grade II degeneration in 4 patients, with a mean LF thickness of 2.43±0.20 mm; grade III in 10 patients, 3.01±0.41 mm; and grade IV in 25 patients, 4.16±1.12 mm. LF thickness significantly increased with grade of disc degeneration and was significantly correlated with age (r=0.55, p<0.01). Logistic regression analysis identified predictive effects of segmental angulation (odds ratio [OR]=1.55, p=0.014) and age (OR=1.16, p=0.008).ConclusionsAge-related increases in disc degeneration, combined with continuous lumbar segmental flexion-extension motion, leads to the development of LF hypertrophy

Analysis of the Prevalence and Distribution of Cervical and Thoracic Compressive Lesions of the Spinal Cord in Lumbar Degenerative Disease

Study DesignRetrospective study.PurposeThe aim of the present study is to analyze the prevalence and distribution of cervical and thoracic compressive lesions of the spinal cord in lumbar degenerative disease, using whole-spine postmyelographic computed tomography.Overview of LiteratureOf the various complications resulting from spinal surgery, unexpected neurological deterioration is the most undesired. There are reports of missed compressive lesions of the spinal cord at the cervical or thoracic level in lumbar degenerative disease.MethodsThere were 145 consecutive patients with symptomatic lumbar degenerative disease evaluated. Before the lumbar surgery, image data were obtained. The following parameters at the cervical and thoracic levels were analyzed: compressive lesions from the anterior parts; compressive lesions from the anterior and posterior parts; ossification of the ligamentum flavum; ossification of the posterior longitudinal ligament; and spinal cord tumor.ResultsCompressive lesions from the anterior parts were observed in 34 cases (23.4%). Compressive lesions from the anterior and posterior parts were observed in 34 cases (23.4%). Lesions of ossification of the ligamentum flavum were observed in 45 cases (31.0%). Lesions of ossification of the posterior longitudinal ligament were observed in 15 cases (10.3%). Spinal cord tumor was not observed.ConclusionsA survey of compressive lesions at the cervical or thoracic level in lumbar degenerative disease is important in preventing unexpected neurological deterioration after the lumbar surgery

Predictable Imaging Signs of Cauda Equina Entrapment in Thoracolumbar and Lumbar Burst Fractures with Greenstick Lamina Fractures

Study DesignA retrospective study.PurposeThe aim of present study was to investigate imaging findings suggestive of cauda equina entrapment in thoracolumbar and lumbar burst fractures.Overview of LiteratureBurst fractures with cauda equina entrapment can cause neurologic deterioration during surgery. However, dural tears and cauda equina entrapment are very difficult to diagnose clinically or radiographically before surgery.MethodsTwenty-three patients who underwent spinal surgery for thoracolumbar or lumbar burst fractures were enrolled in this study. In magnetic resonance imaging T2-weighted images of the transverse plane, we defined cauda equina notch sign (CENS) as a v-shaped image that entrapped cauda equina gathers between lamina fractures. We evaluated the fractured spine by using CENS and lamina fractures and the rate of available space for the spinal canal at the narrowest portion of the burst fracture level. We classified patients into entrapment group or non-entrapment group, based on whether cauda equina entrapment existed.ResultsLamina fractures were detected in 18 (78.3%) and CENS were detected in 6 (26.1%) of 23 burst-fracture patients. Cauda equina entrapment existed in all the patients with CENS. In addition, the rate of available space for the spinal canal increased according to logistic regression. The size of the retropulsed fragment in the spinal canal was the most reliable of all the factors, suggesting cauda equina entrapment.ConclusionsCENS was the most predictable sign of cauda equina entrapment associated with burst fractures

Successful use of dupilumab for egg-induced eosinophilic gastroenteritis with duodenal ulcer: a pediatric case report and review of literature

Background Non-esophageal eosinophilic gastrointestinal disorder (non-EoE-EGID) is a rare disease in which eosinophils infiltrate parts of the gastrointestinal tract other than the esophagus; however, the number of patients with non-EoE-EGID has been increasing in recent years. Owing to its chronic course with repeated relapses, it can lead to developmental delays due to malnutrition, especially in pediatric patients. No established treatment exists for non-EoE-EGID, necessitating long-term systemic corticosteroid administration. Although the efficacy of dupilumab, an anti-IL-4/13 receptor monoclonal antibody, for eosinophilic esophagitis, has been reported, only few reports have demonstrated its efficacy in non-EoE EGIDs. Case presentation A 13-year-old boy developed non-EoE-EGID with duodenal ulcers, with chicken eggs as the trigger. He was successfully treated with an egg-free diet, proton pump inhibitors, and leukotriene receptor antagonists. However, at age 15, he developed worsening upper abdominal pain and difficulty eating. Blood analysis revealed eosinophilia; elevated erythrocyte sedimentation rate; and elevated levels of C-reactive protein, total immunoglobulin E, and thymic and activation-regulated chemokines. Upper gastrointestinal endoscopy revealed a duodenal ulcer with marked mucosal eosinophilic infiltration. Gastrointestinal symptoms persisted even after starting systemic steroids, making it difficult to reduce the steroid dose. Subcutaneous injection of dupilumab was initiated because of comorbid atopic dermatitis exacerbation. After 3 months, the gastrointestinal symptoms disappeared, and after 5 months, the duodenal ulcer disappeared and the eosinophil count decreased in the mucosa. Six months later, systemic steroids were discontinued, and the duodenal ulcer remained recurrence-free. The egg challenge test result was negative; therefore, the egg-free diet was discontinued. Blood eosinophil count and serum IL-5, IL-13, and eotaxin-3 levels decreased after dupilumab treatment. The serum levels of IL-5 and eotaxin-3 remained within normal ranges, although the blood eosinophil counts increased again after discontinuation of oral prednisolone. Conclusions Suppression of IL-4R/IL-13R-mediated signaling by dupilumab may improve abdominal symptoms and endoscopic and histologic findings in patients with non-EoE-EGID, leading to the discontinuation of systemic steroid administration and tolerance of causative foods