Search

Aims: This study aimed to determine whether pathological changes in the bone marrow cause Osteoarthritis (OA) pain based on magnetic resonance imaging (MRI), immunohistochemistry, and electrophysiology., Main Methods: Adjuvant-induced arthritis (AIA) was achieved by injecting 150 μL of complete Freund's adjuvant into the right knee joints of male Sprague-Dawley rats. AIA rats were compared with saline-injected rats., Key Findings: AIA significantly induced mechanical hyperalgesia and spontaneous pain in the right hind paw 1-14 days after induction. Intratibial injection of 50 μL of 1 % lidocaine significantly suppressed AIA-induced mechanical hyperalgesia (p = 0.0001) and spontaneous pain (p = 0.0006) 3 days after induction. In T2-weighted MRI, AIA induced high-signal intensity within the proximal tibial metaphysis, and the mean T2 values in this area significantly increased on days 3 (p = 0.0043) and 14 (p = 0.0012) after induction. AIA induced intraosseous edema and significantly increased the number of intraosseous granulocytes on days 3 (p < 0.0001) and 14 (p < 0.0001) after induction. The electrophysiological study on days 3-7 after induction showed significantly increased spontaneous firing rates (p = 0.0166) and evoked responses to cutaneous stimuli (brush, p < 0.0001; pinching, p = 0.0359) in the right hind paw plantar surface and intratibial stimuli (p = 0.0002) in wide-dynamic-range neurons of the spinal dorsal horn., Significance: Intraosseous changes caused by OA induce hypersensitivity in the sensory afferents innervating bone marrow may be involved in OA pain. Novel bone marrow-targeted therapies could be beneficial for treating OA pain., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Dengue is a febrile illness caused by the dengue virus (DENV) that belongs to the genus Flavivirus in the family Flaviviridae. Cross-reactivity between flaviviruses poses a challenge while interpreting serological test results. In the present study, the cross-reactivity of sera of the patients with dengue, who traveled from Japan to DENV-endemic countries, was analyzed by using an enzyme-linked immunosorbent assay (ELISA) and neutralization test (NT). Sixteen serum samples were collected from patients with dengue and were tested for: i) IgM antibodies against Zika virus (ZIKV), West Nile virus (WNV), Japanese encephalitis virus (JEV), and tick-borne encephalitis virus (TBEV) using IgM ELISA, ii) IgG antibody against TBEV using IgG ELISA, and iii) neutralizing antibody against ZIKV, WNV, TBEV, and JEV. Among the 16 samples tested using ELISA, seven samples were IgM-positive for at least one of the other flaviviruses, and nine samples were IgG-positive for TBEV. Neutralizing antibody titers (NATs) against ZIKV, WNV, and TBEV were one-fourth or lower than those against the causative DENV in all samples. The NATs against JEV were one-fourth or lower than those against the causative DENV in six convalescent-phase serum sample among the seven convalescent-phase serum samples. The NAT against DENV of the residual one convalescent-phase serum was similar to that against JEV and that against JEV of its relevant acute-phase serum sample. These results showed that NTs with paired serum samples are important to correctly interpret the serological test results for DENV., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

The Chikungunya virus (CHIKV), an enveloped RNA virus that has been identified in over 40 countries and is considered a growing threat to public health worldwide. However, there is no preventive vaccine or specific therapeutic drug for CHIKV infection. To identify a new inhibitor against CHIKV infection, this study constructed a subgenomic RNA replicon expressing the secretory Gaussia luciferase (Gluc) based on the CHIKV SL11131 strain. Transfection of in vitro-transcribed replicon RNA to BHK-21 cells revealed that Gluc activity in culture supernatants was correlated with the intracellular replication of the replicon genome. Through a chemical compound library screen using the Gluc reporter CHIKV replicon, we identified several compounds that suppressed CHIKV infection in Vero cells. Among the hits identified, CP-154,526, a non-peptide antagonist of the corticotropin-releasing factor receptor type-1 (CRF-R1), showed the strongest anti-CHIKV activity and inhibited CHIKV infection in Huh-7 cells. Interestingly, other CRF-R1 antagonists, R121919 and NGD 98-2, also exhibited inhibitory effects on CHIKV infection. Time-of-drug addition and virus entry assays indicated that CP-154,526 suppressed a post-entry step of infection, suggesting that CRF-R1 antagonists acted on a target in the intracellular replication process of CHIKV. Therefore, the Gluc reporter replicon system established in this study would greatly facilitate the development of antiviral drugs against CHIKV infection., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Youichi Suzuki reports financial support was provided by Japan Agency for Medical Research and Development., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: Few studies have examined a possible correlation between cervical disc degeneration and disc height. The aim of this study was to establish age-related changes and sex-specific differences of cervical disc height using magnetic resonance imaging (MRI) and to evaluate the relationship between degree of cervical disc degeneration and disc height in asymptomatic subjects., Methods: We measured the intervertebral disc anteroposterior diameter and disc height in each disc level using MRI in 1211 relatively healthy volunteers (606 men and 605 women, mean age 49.5 years). We included at least 100 males and 100 females in each decade of life between the 20s and the 70s. Cervical disc degeneration was defined according to the modified Pfirrmann classification system, and disc index and intervertebral disc height narrowing ratio were evaluated on sagittal plane MRI., Results: Intervertebral disc height decreased gradually with increasing age in both sexes. The grade of disc degeneration significantly increased with age in both sexes at every level. Mild disc degeneration was observed even in subjects their 20s. Disc degeneration occurred around the C5-C6 level. The average disc height decreased with aging after the 40s. Disc index and intervertebral disc height narrowing ratio decreased with a progression in the disc degeneration grade in both sexes., Conclusions: This large-scale cross-sectional analysis of cervical spine MRI data in healthy subjects demonstrated that cervical disc height narrowing progresses with age. Cervical disc height narrowing is highly correlated with the progression of disc degeneration., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: Postoperative SSI is a common and potentially serious complication in spine surgery. Seasonal variation occurs in rates of nosocomial infection, with higher rates found in the summer, during which hot, humid conditions may be optimal for proliferation of bacteria. This might also influence the rate of SSI. The purpose of the study was to examine seasonal variation in SSI after PLIF/TLIF surgery, including relationships with experience of surgeons and causal organisms., Methods: Cases with SSI after PLIF/TLIF surgery at 10 facilities between January 1, 2012, and December 31, 2014 were retrieved from a database. Infection was defined based on CDC guidelines for SSIs. Patients were followed for at least two years after surgery. Surgeries were examined in spring (April-June), summer (July-September), autumn (October-December), and winter (January-March). Seasonal variation and other factors with a potential association with SSIs were evaluated., Results: A total of 1174 patients (607 males, 567 females) who underwent PLIF/TLIF surgery were identified. The operations were PLIF (n = 667), TLIF (n = 443), MIS-PLIF (n = 27), and MIS-TLIF (n = 37). The total SSI rate for the 2-year period was 2.5% (29/1174), and the 2-year average SSI rates for surgeries in each season were spring, 2.6% (7/266); summer, 3.9% (13/335); fall, 1.3% (4/302); winter, 1.8% (5/271). The SSI rate was significantly higher in summer than non-summer (3.9% vs. 1.9%, p < 0.05). SSIs were caused by a variety of pathogens, including Gram-positive cocci, and Staphylococcus aureus was the most common pathogenic organism to cause SSI., Conclusion: Seasonality should be taken into account in strategies for SSI prevention, with particular attention on mitigation of increased temperature and humidity in the summer and on infection caused by Gram-positive cocci and S. aureus., Competing Interests: Declaration of Competing Interest None of the authors have a conflict of interest., (Copyright © 2020 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Background: The mechanism of vascular remodelling in pulmonary arterial hypertension (PAH) remains unclear. Hence, defining the origin of cells constituting intractable vascular lesions in PAH is expected to facilitate therapeutic progress. Herein, we aimed to evaluate the origin of intractable vascular lesions in PAH rodent models via bone marrow (BM) and orthotopic lung transplantation (LT)., Methods: To trace BM-derived cells, we prepared chimeric rats transplanted with BM cells from green fluorescent protein (GFP) transgenic rats. Male rats were transplanted with lungs obtained from female rats and vice versa. Pulmonary hypertension was induced in the transplanted rats via Sugen5416 treatment and subsequent chronic hypoxia (Su/Hx)., Results: In the chimeric Su/Hx models, GFP-positive cells were observed in the pulmonary vascular area. Moreover, the right ventricular systolic pressure was significantly lower compared with wild-type Su/Hx rats without BM transplantation (P = 0.009). PAH suppression was also observed in rats that received allograft transplanted BM transplantation. In male rats that received LT and Su/Hx, BM-derived cells carrying the Y chromosome were also detected in neointimal occlusive lesions of the transplanted lungs received from female rats., Conclusions: BM-derived cells participate in pulmonary vascular remodelling in the Su/Hx rat model, whereas BM transplantation may contribute to suppression of development of PAH., (Copyright © 2021 Canadian Cardiovascular Society. Published by Elsevier Inc. All rights reserved.)

Objectives: This study aimed to evaluate the anteroposterior diameters and cross-sectional areas of the dural sac and spinal cord in the thoracic spine, to elucidate the spinal cord occupation rate of the dural sac in these dynamic changes for each level using multidetector-row computed tomography (MDCT)., Patients and Methods: Fifty patients with cervical or lumbar spinal disease were prospectively enrolled. After preoperative myelography, MDCT was performed at maximum passive flexion and extension. The anteroposterior diameter and cross-sectional area of the dural sac and spinal cord in the axial plane and the thoracic spinal cord length in the sagittal plane were measured. The spinal cord occupation rate in the dural sac was calculated., Results: The spinal cord occupation rate of the dural sac in anteroposterior diameter was lower on flexion than on extension, with significant differences from the T1/T2 to T11/T12 levels (p < 0.0001). The spinal cord occupation rate of the dural sac in cross-sectional area was lower on flexion than on extension, with significant differences except from T3/T4 to T6/T7 levels (p < 0.01). There was a bimodal increase in the occupation rate with elevations at the cervicothoracic junction and thoracolumbar junction. The thoracic spinal cord length on flexion was significantly longer than that on extension (p < 0.0001)., Conclusions: The spinal cord occupation rate of the dural sac was lower on flexion than on extension, despite thoracic spine being considered a rigid region. The dynamic changes in longitudinal stretching and shrinkage of the spinal cord affected the occupation rate., (Copyright © 2020. Published by Elsevier B.V.)

Background: Pulmonary endothelial damage has a negative impact on the maintenance of normal pulmonary vascular function. Such damage results in delayed thrombus dissolution and vascular remodeling in chronic thromboembolic pulmonary hypertension (CTEPH). Although endothelial progenitor cells (EPCs) may be incorporated into neovasculature during vascular repair, their function in CTEPH remains unclarified, especially under the augmentation of soluble guanylate cyclase (sGC) activity., Methods and Results: We evaluated the effect of EPCs on endothelial function and compared the effect of riociguat, a sGC stimulator, on the number and function of circulating EPCs in two groups of CTEPH patients. The two groups consisted 16 CTEPH patients who were treatment naïve (Naïve group), and 14 CTEPH patients who were being treated with riociguat, a sGC stimulator (Riociguat group). The number of circulating EPCs in the Riociguat group was significantly higher than that in the Naïve group. Gene expression levels associated with angiogenesis were significantly higher in EPCs of the Riociguat group. EPC-stimulated tube formation and migration of human pulmonary microvascular endothelial cell (hPMVEC) in the Riociguat group exceeded that in the Naïve group. The angiogenic ability of hPMVECs stimulated by EPCs in the Riociguat group was enhanced compared to that of the sGC stimulator, BAY 41-2272., Conclusion: These findings indicate that riociguat may induce EPCs to play a protective role via modulation of endothelial functions associated with CTEPH., Translation Aspect of the Work: Endothelial dysfunction exacerbates CTEPH. Riociguat enhanced the protective role of EPCs via neovascularization, which prevented vascular remodeling and alleviated CTEPH., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Optimal treatment of lumbar spinal stenosis (LSS) with neurological deficit due to osteoporotic vertebral fractures (OVFs) has been controversial. We assessed the usefulness, safety, and efficacy of posterior lumbar interbody fusion (PLIF) for LSS with neurological deficit due to OVFs and compared this procedure to posterior/anterior combined surgery (PACS)., Methods: Of 36 consecutive patients with LSS with neurological deficit due to OVFs, 15 underwent PLIF (6 males, 9 females; mean age, 74 years), and 21 underwent PACS (4 males, 17 females; mean age, 70 years). Surgical complications, clinical outcomes (operative time, blood loss, American Spinal Injury Association Impairment Scale [AIS], activities of daily living [ADLs]), and sagittal alignment were investigated. Bony fusion was assessed using plain and functional X-rays and computed tomography scans., Results: There were no significant differences in age, sex, or disease or follow-up duration between the groups. Operative time was significantly shorter and intraoperative blood loss significantly less in the PLIF than in the PACS groups. AIS and ADL improved significantly postoperatively in both groups. No significant difference was observed in neurological improvement, correction angle, loss of correction, and surgical complications. No pseudarthrosis occurred, and no patient required additional surgery in the PLIF group., Conclusions: PLIF for LSS with neurological deficit due to OVFs achieves posterior rigid fixation with instrumentation, anterior column reconstruction by interbody fusion, and adequate decompression using a single posterior approach. This less invasive procedure is a useful reconstructive surgery option., (Copyright © 2019 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Dengue virus (DENV) is the causative agent of dengue fever (DF), dengue haemorrhagic fever (DHF), and dengue shock syndrome (DSS) and continues to be a public health problem in the tropical and subtropical areas. However, there is currently no antiviral treatment for DENV infection. In this study, our aim was to develop a stable reporter replicon cell system that supports constant viral RNA replication in cultured cells. The isolated replicon cells exhibited high levels of luciferase activity in the culture supernatant concomitant with expression of virus-encoded NS1, NS3 and NS5 proteins in the cells. The NS1, NS3 proteins and dsRNA were detected in the replicon cells by immunofluorescence analysis. Furthermore, the anti-DENV inhibitors ribavirin and bromocriptine significantly reduced the luciferase activity in a dose-dependent manner. High-throughput screening with a compound library using the stably-transfected replicon cells showed a Z' factor value of 0.57. Our screening yielded several candidates including one compound that has already shown anti-DENV activity. Taken together, our results demonstrate that this DENV subgenomic replicon cell system expressing a secretory luciferase gene can be useful for the high-throughput screening of anti-DENV compounds and the analysis of the replication mechanism of the DENV RNA., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Japanese encephalitis (JE) is one of the most important viral encephalitis in Asia. JE is caused by the Japanese encephalitis virus (JEV), which belongs to the genus Flavivirus, family Flaviviridae. The diagnosis of JE is usually based on serological assays, and it has been reported that cross-reactivity between flaviviruses has complicated the interpretations of results from serological assays. Therefore, analysis of the cross-reactivity is an important subject for serological diagnosis of JE and other diseases caused by flaviviruses. In the present study, the cross-reactivity of the sera of patients with JE to other flaviviruses was analyzed using enzyme-linked immunosorbent assay (ELISA) and neutralization tests. Sixteen serum samples were collected from patients with JE and were tested for: i) IgM antibody against West Nile virus (WNV), dengue virus (DENV), zika virus (ZIKV), and tick-borne encephalitis virus (TBEV) using IgM-ELISA, ii) IgG antibody against DENV and TBEV using IgG-ELISA, and iii) neutralization tests with DENV 1-4, ZIKV, TBEV, and WNV. Out of the 16 samples tested using ELISA, 11 and 14 samples were positive for IgM and IgG, respectively, against at least one of the other flaviviruses. In neutralization tests, neutralizing potency against DENV, ZIKV, or TBEV was not detected in any samples. Although 13 samples showed neutralizing potency against WNV, their neutralizing antibody titers were equal to or less than one-eighth of those against JEV. These results show that neutralization tests are more specific than ELISA, indicating the importance of the neutralization tests in the diagnosis of JE., (Copyright © 2019 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Objectives: The purpose of this study was to establish the clinical cut-off values of the 10-s grip and release (G&R) and 10-s step quantitative tests for the diagnosis of cervical spondylotic myelopathy (CSM) and to elucidate the aging variation and gender difference of those values in a large cohort of healthy subjects., Patients and Methods: Patients with CSM (n = 454) and asymptomatic subjects (n = 818) were included. Subjects were aged 40-70 years; according to their age, they were categorized by decades. The 10-s G&R and 10-s step tests were used to quantitatively assess performance. The receiver operating characteristic (ROC) curves were plotted to evaluate the cut-off value of the 10-s G&R and 10-s step tests for determining the presence or absence of CSM in each gender and decade., Results: The cut-off values in the G&R test were 20 in 40 s, 19 in 50 s, 17 in 60 s, and 16 in 70 s groups respectively. The cut-off values in the 10-s step test were 19 in 40 s, 18 in 50 s, 16 in 60 s, and 15 in 70 s groups respectively. The cut-off value of the G&R test in females was lower than that in the males. The cut-off value of 10-s step test was lower in the females than in the males in 40 s and 50 s groups., Conclusions: The cut-off values in the 10-s G&R test and 10-s step decreased with age. When these quantifiable tests are used as screening tests of CSM, age and gender difference should be considered., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Dengue virus (DENV) is the most prevalent human arthropod-borne virus and causes severe problems worldwide, mainly in tropical and sub-tropical regions. However, there is no specific antiviral drug against DENV infection. We and others recently reported that stearoyl-CoA desaturase-1 (SCD1) inhibitor showed potent suppression of hepatitis C virus replication. In this study, we examined the impact of SCD1 on DENV replication. We found that SCD1 inhibitors (MK8245 and #1716) dramatically suppressed DENV replication in a dose-dependent manner without cytotoxicity. This anti-DENV efficacy was observed against all four DENV serotypes and other flaviviruses, including Zika virus and Japanese encephalitis virus. A subgenomic replicon system of DENV was used to confirm that SCD1 inhibitor suppressed viral RNA replication. Interestingly, exogenous supplementation of unsaturated fatty acids resulted in recovery of the DENV titer even in the presence of SCD1 inhibitor, suggesting that fatty acid biosynthesis contributes to DENV genome replication. These findings indicate that SCD1 is a novel host factor required for DENV replication, and SCD1 inhibitor is a potential candidate for treating dengue fever., (Copyright © 2019. Published by Elsevier B.V.)

Objectives: To identify factors associated with prolonged length of stay (LOS) in spine surgery, with the goal of establishing details of LOS for multiple diseases and surgical procedures., Patients and Methods: The subjects were patients who underwent spine surgery at 10 facilities in the Nagoya Spine Group from January 2005 to December 2015. Data were collected for patient background, primary spinal pathology, anatomical location of the lesion, and surgical methods. The primary outcome was LOS, which was defined as the calendar days from surgery to hospital discharge., Results: A total of 10,829 patients (5953 males, 4876 females; age 5-93 years, mean 60.2 ± 28.8 years) were identified in the database. Average follow-up was 61 months (range: 13-120 months). Average LOS was 22.3 ± 21.3 days, and there was a gradual decrease in LOS over the study period. LOS was significantly correlated with age, and prolonged LOS was significantly associated with thoracic spine surgery and significantly longer after surgery with instrumentation. Average LOS was >30 days for intramedullary tumor resection and posterior cervical fusion, but only 10.2 days for microendoscopic discectomy. Reoperation was performed in 210 patients (1.9%) and these patients had a significantly higher average LOS of 43.1 days., Conclusion: These results will assist quality improvement in spine surgery. The identified risk factors for prolonged LOS will also assist in planning of surgery, postoperative care, and discharge, with the goal of reducing health care costs., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objectives: Although patients with cervical spinal cord injury without radiographic abnormality (SCIWORA) present increased signal intensity (ISI) on magnetic resonance imaging (MRI), its degree has not been examined. This study evaluated the clinical effectiveness of MRI-based ISI in adult patients of SCIWORA. Its predictive value for symptom severity was also evaluated., Patients and Methods: One-hundred consecutive SCIWORA patients who had undergone expansive laminoplasty were enrolled. Among them, 79 were male and 21 were female. The mean age was 55 years (range 20-87). All patients underwent MRI in the acute phase, and ISI was classified into three groups based on sagittal T2-weighted MRI: Grade 0, none; Grade 1, light (obscure); and Grade 2, intense (bright). The pre- and postoperative neurological status was evaluated using the Japanese Orthopaedic Association scoring system for cervical myelopathy (JOA score) and the ASIA impairment scale (AIS)., Results: Preoperative MRI showed Grade 0 in 8 patients, Grade 1 in 49 patients, and Grade 2 in 43 patients. There were no differences in age and gender among three groups. The pre- and postoperative JOA scores decreased significantly with an increasing ISI grade. The recovery rate of JOA score decreased with the ISI grade. The ISI grade tended to increase with the pre- and postoperative AIS grades. ISI Grade 2 on MRI was observed in severely paralyzed cases., Conclusions: MRI-based ISI classification is correlated with preoperative symptom severity in adult patients with SCIWORA and can be a predictor of surgical outcome., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: The goals of the study are to analyze postoperative outcomes and recurrence in cases of spinal cord and cauda equina ependymoma in each World Health Organization (WHO) Grade, and to examine the influence of extent of surgical removal on prognosis. Spinal ependymoma has a relatively high frequency among intramedullary spinal cord tumors. The tumor is classified in WHO guidelines as grades I, II, and III, but few studies have examined postoperative prognosis based on these grades., Methods: The records of 80 patients undergoing surgery for spinal cord and cauda equina ependymoma were examined in a multicenter study using a retrospective database. Neurological motor status, pathological type, extent of resection, and tumor recurrence were evaluated., Results: The histopathological types were grade I in 23 cases (myxopapillary: 21, subependymoma: 2), grade II in 52 cases, and grade III in 5 cases (including all anaplastic cases). Total resection was performed in 60 cases (83%), and eight cases had recurrence, including 2 in WHO grade I, 2 in grade II, and 4 in grade III. The 5-year recurrence-free survival rates were 90%, 91%, and 20% in grades I, II and III, respectively. Adjuvant radiotherapy for the local site was performed in 8 cases, including 3 in grade I and 5 in grade III; however, 4 of the 5 grade III cases (80%) had recurrence after radiotherapy. Among 59 patients with normal ambulation or independence without external assistance (McCormick Grade I or II), 53 (90%) maintained the same mobility after surgery. In cases that underwent total resection, the recurrence rate was significantly lower (p < 0.01). A good preoperative motor status also resulted in significantly better postoperative recovery of motor status (p < 0.05)., Conclusions: Total resection of spinal cord and cauda equina ependymoma leads to postoperative motor recovery and may reduce tumor recurrence. Therefore, early surgery for this tumor is recommended before aggravation of paralysis., (Copyright © 2018 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Background: Riociguat, the first approved drug for patients with chronic thromboembolic pulmonary hypertension (CTEPH), is a soluble guanylate cyclase (sGC) Stimulator. It directly stimulates sGC independently of nitric oxide (NO) and increases sGC sensitivity for NO. The safety and efficacy of transitioning from a phosphodiesterase 5 inhibitor (PDE5i) to riociguat is unknown., Methods and Results: Twenty-three patients were prospectively enrolled: 8 symptomatic patients with inadequate clinical responses to PDE5i were changed to riociguat (transitioned group); 15 started riociguat anew (new or add-on group). We analyzed the change from baseline to 6-12 months of riociguat treatment for the 6-minute walk distance (6MWD), mean pulmonary arterial pressure (mPAP), pulmonary vascular resistance (PVR), cardiac index (CI), partial pressure of oxygen in arterial blood (PaO2), brain natriuretic peptide (BNP), World Health Organization (WHO) functional class, safety and adverse events. The mPAP, BNP and WHO functional class significantly improved in total. In the transitioned group, BNP significantly decreased by -116.5±188.6pg/ml (P=0.0156). The 6MWD, mPAP, PVR, CI, and PaO2 improved but not significantly. The baseline condition was significantly more severe in the transitioned than in the new or add-on group. No patients discontinued riociguat. Relatively rapid transitioning from PDE5i to riociguat was safe under careful observation., Conclusions: Transitioning to riociguat may be safe and effective in CTEPH patients with inadequate clinical responses to PDE5i., (Copyright © 2017 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.)

Objective: To provide insight into the wide spectrum of migraine during childhood to establish practical and comprehensive treatment strategies., Background: Although recent studies have confirmed the effect of anti-migraine agents in childhood headaches fulfilling the criteria of migraine without aura, there have been no studies regarding the efficacy of these drugs in childhood migraine without aura not filling the diagnostic criteria., Methods: In total, 154 patients with a clinical diagnosis of migraine, with onset of repetitive headaches at the age of ⩽15years, were retrospectively included from clinics in seven tertiary medical centers., Results: Patients' diagnoses included migraine with aura (n=49), migraine without aura (n=65), clinical migraine without aura not fulfilling International Classification of Headache Disorders-3 beta criteria (suspected migraine without aura; n=38), and hemiplegic migraine (n=2). Abortive medicine was effective in 74 of 97 patients, and preventive medicine was effective in 61 of 84 patients. Drugs with high efficacy were acetaminophen and ibuprofen for abortive therapy and cyproheptadine, amitriptyline, and propranolol for preventive therapy. Psychosocial problems were less common, and abnormalities on electroencephalography were more common in the suspected migraine without aura group. Otherwise, clinical features and drug responsibility were comparable among the migraine with aura, migraine without aura, and suspected migraine without aura groups. Retrospectively, experts clinically diagnosed childhood migraine without aura when the headache met at least one of the three criteria B, C, and D in International Classification of Headache Disorders-3 beta in addition to A and E. Abortive and preventive medication including paroxetine (n=2) benefited 10 and 15 of the 33 patients with daily headache, respectively. Psychotherapy/counseling (n=4), treatment for orthostatic dysregulation (n=4), and elimination of stressors (n=3) markedly alleviated headache in this group., Conclusion: Our results indicated that those with suspected migraine without aura not filling International Classification of Headache Disorders diagnostic criteria should be included in the treatment for migraine. Treatment should also be targeted to comorbid developmental disorders, orthostatic dysregulation, and psychosocial problems in patients with refractory daily headaches., (Copyright © 2016 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

Objectives: To examine the clinical efficacy of intrathecal morphine as postoperative analgesia for cervical laminoplasty., Summary of Background Data: Patients who undergo posterior cervical spinal surgery frequently experience significant postoperative pain. Postoperative pain contributes to patient morbidity because of decreasing early voluntary mobilization and delayed rehabilitation. Intrathecal morphine is known to be a simple and effective analgesia. However, the effectiveness of intrathecal morphine for cervical spinal surgery has not yet been reported., Methods: Seventy-eight patients with cervical spondylotic myelopathy were divided into two groups prospectively, a diclofenac suppository (DS) group who received 50 mg diclofenac suppository at the end of the surgery, and an intrathecal morphine (ITM) group who were preoperatively administered 0.3 mg of morphine chloride, intrathecally, via a lumbar puncture. All patients underwent double-door laminoplasty of C3-6 or C3-7 level. Visual analog scale (VAS) of cervical pain, self-rating pain impression, supplemental analgesic usage, and complication rate were evaluated until the seventh postoperative day., Results: Thirty-one patients in the DS group and 32 patients in the ITM group were finally assessed. No baseline variable differences between the two groups were observed. The VAS was significantly lower in the ITM group at 4 h and 24 h until the seventh postoperative day. Self-rating pain impression was significantly better in the ITM group. No significant difference was observed in complication rate., Conclusions: Intrathecal morphine was an effective and safe analgesic method for cervical laminoplasty in patients with cervical spondylotic myelopathy., (Copyright © 2016 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Dengue virus (DENV) infectious disease is a major public health problem worldwide; however, licensed vaccines or specific antiviral drugs against this infection are not available. To identify novel anti-DENV compounds, we screened 1280 pharmacologically active compounds using focus reduction assay. Bromocriptine (BRC) was found to have potent anti-DENV activity and low cytotoxicity (half maximal effective concentration [EC50], 0.8-1.6 μM; and half maximal cytotoxicity concentration [CC50], 53.6 μM). Time-of-drug-addition and time-of-drug-elimination assays suggested that BRC inhibits translation and/or replication steps in the DENV life cycle. A subgenomic replicon system was used to verify that BRC restricts RNA replication step. Furthermore, a single amino acid substitution (N374H) was detected in the NS3 protein that conferred resistance to BRC. In summary, BRC was found to be a novel DENV inhibitor and a potential candidate for the treatment of DENV infectious disease., (Copyright © 2016 The Author(s). Published by Elsevier B.V. All rights reserved.)

Much attention has been paid to cholestane-type steroidal glycosides because of their importance from the perspectives of both chemical diversity and significant biological activities. A phytochemical investigation of the rhizomes of Polygonatum odoratum (Liliaceae) resulted in the isolation of three novel cholestane-type steroidal glycosides (1-3) with unique Δ(14,16)-unsaturated D-ring structures as well as two novel spirostane-type steroidal saponins (4 and 5) and three known steroidal glycosides (6-8). Their structures were determined by various spectroscopic methods and chemical reactions. Steroidal saponin 7 showed significant antifungal activity against Candida albicans JCM1542 (MIC 3.1 μg/mL) and Aspergillus fumigatus JCM1738 (MIC 6.3 μg/mL)., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Purpose: The 10-s grip and release (G&R) test and 10-s step test were reported to be useful tools to evaluate the severity of cervical myelopathy quantitatively. The purpose of this study is to establish the standard values of the 10-s G&R test and 10-s step test as quantitative tests for cervical myelopathy and to elucidate the aging variation and gender difference of those values in a large cohort of normal subjects., Methods: A total of 1230 healthy volunteers were enrolled. They included at least 100 men and 100 women in each decade from the 20s to 70s. Three tests were performed: the number of finger grips and releases in 10 s, number of steps in 10 s, and gripping power., Results: The average number of G&Rs on the weaker side was 21.5 ± 5.5, whereas the average number of steps was 19.7 ± 3.4. The average gripping power on the weaker side was 29.5 ± 9.6 kg. The number of G&Rs and steps in 10 s decreased significantly with age (r = -0.58, -0.43, respectively). The average number of G&Rs on the weaker side was <20 in the 60s and 70s. The number of G&Rs was significantly correlated with the number of steps and gripping power. A significant difference was seen between the genders in all three tests., Conclusion: The standard values of the 10-s G&R test, 10-s step test, and gripping power were established in this study. When these quantifiable tests are used as screening tests of cervical myelopathy, aging variation and gender difference should be considered.

Background: The classification and therapeutic strategy for thoracolumbar burst fractures are controversial. The load-sharing classification (LSC) and thoracolumbar injury classification system (TLICS) are both quantitative evaluation systems for thoracolumbar burst fractures. We hypothesized that their combination would be helpful not only for surgical indications but also for deciding on the surgical approach. However, no reports have evaluated the relationship between them. The purpose of this study was to clarify the relationship between the LSC and TLICS and investigate the clinical usefulness of their combination., Methods: This study included 100 consecutive patients surgically treated for thoracolumbar burst fractures (71 men and 29 women; mean age 36 years). Clinical and radiographical data as well as thoracolumbar injury classification systems were evaluated., Results: LSC and TLICS scores were found to be statistically correlated. The mean LSC score with a TLICS score of 5 or more (surgical treatment recommended) was 7.3 ± 1.2 points, and the mean LSC score with a TLICS score of 3 or less (conservative treatment recommended) was 6.1 ± 1.3 points. The mean TLICS score with an LSC score of 7 or more (additional anterior reconstruction recommended) was 6.6 ± 2.7 points, and the mean TLICS score with an LSC score of 6 or less (expectation of good clinical results with posterior short fusion) was 5.0 ± 2.5 points. The TLICS score was 3 or less, and the LSC score was 7 or more in 13 patients (13 %)., Conclusion: Although the TLICS scores correlated with the LSC scores, a single application of TLICS might not be sufficient to identify those patients who have a TLICS score of 3 or less and an LSC score of 7 or more as surgically indicated. However, an additional LSC evaluation avoided deviations as the two classifications complemented each other, and it was useful in determining the best treatment options for thoracolumbar burst fractures.

O-linked methylation of sugar substituents is a common modification in the biosynthesis of many natural products and is catalyzed by multiple families of S-adenosyl-L-methionine (SAM or AdoMet)-dependent methyltransferases (MTs). Mycinamicins, potent antibiotics from Micromonospora griseorubida, can be methylated at two positions on a 6-deoxyallose substituent. The first methylation is catalyzed by MycE, a SAM- and metal-dependent MT. Crystal structures were determined for MycE bound to the product S-adenosyl-L-homocysteine (AdoHcy) and magnesium, both with and without the natural substrate mycinamicin VI. This represents the first structure of a natural product sugar MT in complex with its natural substrate. MycE is a tetramer of a two-domain polypeptide, comprising a C-terminal catalytic MT domain and an N-terminal auxiliary domain, which is important for quaternary assembly and for substrate binding. The symmetric MycE tetramer has a novel MT organization in which each of the four active sites is formed at the junction of three monomers within the tetramer. The active-site structure supports a mechanism in which a conserved histidine acts as a general base, and the metal ion helps to position the methyl acceptor and to stabilize a hydroxylate intermediate. A conserved tyrosine is suggested to support activity through interactions with the transferred methyl group from the SAM methyl donor. The structure of the free enzyme reveals a dramatic order-disorder transition in the active site relative to the S-adenosyl-L-homocysteine complexes, suggesting a mechanism for product/substrate exchange through concerted movement of five loops and the polypeptide C-terminus., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Macrolides are a class of valuable antibiotics that include a macrolactone ring, at least one appended sugar unit, and, in most cases, additional hydroxyl or epoxide groups installed by cytochrome P450 enzymes. These functional groups contribute to structural diversification and serve to improve the bioactivity profiles of natural products. Here, we have characterized in vitro two P450 enzymes from the mycinamicin biosynthetic pathway of Micromonospora griseorubida. First, MycCI was characterized as the C21 methyl hydroxylase of mycinamicin VIII, the earliest macrolide form in the postpolyketide synthase tailoring pathway. Moreover, we established that optimal activity of MycCI depends on the native ferredoxin MycCII. Second, MycG P450 catalyzes consecutive hydroxylation and epoxidation reactions with mycinamicin IV as initial substrate. These reactions require prior dimethylation of 6-deoxyallose to mycinose for effective conversion by the dual function MycG enzyme.

We report on a boy with bilateral optic nerve hypoplasia and mild psychomotor retardation. At 1 year and 9 months of age, he was admitted to hospital with a cluster of febrile convulsions and unconsciousness. Magnetic resonance imaging (MRI) revealed widespread areas of high signal intensity on diffusion-weighted imaging of the deep cerebral white matter and corpus callosum. This imaging disappeared at five days of illness. No atrophy or abnormalities were noted on the 6-month follow-up MRI. Despite full recovery after the acute episode, the patient showed retarded developmental progress. We discuss the differential diagnosis for this case.

Introduction of antibodies which retain their function into cells using a simple and easy method would be very useful for study of the intracellular events in living cells. In this study, we developed a new method for intracellular delivery of antibody. It includes a combination of a novel IgG-capturing protein and hemagglutinating virus of Japan envelope (HVJ-E), an inactivated Sendai virus particle which can deliver a variety of molecules into mammalian cells via membrane-fusing activity. The IgG-capturing protein, which was molecularly designed to have two functions, was prepared as a fusion protein (ZZ-NP) of ZZ-dimer derived from an immunoglobulin-binding domain of protein A and nucleocapsid protein (NP), a part of the structural protein of HVJ. ZZ-NP was efficiently incorporated into the HVJ-E particle by treatment with detergent, and enhanced the incorporation of IgG. Moreover, fluorescence immunostaining revealed that the incorporated antibody was very efficiently introduced into living cells while retaining its function, i.e. anti-NPC (nuclear pore complex) monoclonal antibody was selectively located around cell nuclei. These findings suggest that this method is useful for intracellular delivery of antibody and for analysis of biological function of sub-cellular molecules in living cells.

To explore clinical protocols for the prevention of early-onset group B Streptococcus (EOGBS) disease of the newborn in Japan, we conducted a multicenter questionnaire survey. Of 32 regional centers participating in the Kyoto Neonatal Study Group, 28 provided usable data concerning prevention practices undertaken between 2000 and 2004. Twenty-three (82%) of the 28 hospitals implemented bacteriological screening to identify maternal GBS carriage, and all 23 hospitals administered intrapartum antibiotics to all screening-positive pregnant women. There were no institutes that used risk-based strategies. In the 23 hospitals, bacteriological screening was conducted mostly by lower vaginal swab alone (n = 18). Eighteen hospitals performed screening once during pregnancy, either before 34 weeks' gestation (n = 6) or between 35 and 37 weeks' gestation (n = 12). Oral antepartum antibiotics, when carriage was identified, were administered at 12 (52%) hospitals. Twenty institutes used penicillins for intrapartum prophylaxis. However, the loading dose for chemoprophylaxis ranged from 0.5 to 2 g, and the interval between repeat administrations ranged from 4 to 12 h. Although the results indicated that more than 80% of the hospitals surveyed had introduced some screening-based prevention practices, the timing of the bacteriological screening during the pregnancy, the number of screenings, and the screening sites, as well as the antibiotics used, and their dosage, varied widely. Because of these highly variable methods, the efficacy of the implementation of preventive practices could not be determined. This study is the first to have described preventive practices for EOGBS disease in Japan in the era of Centers for Disease Control and Prevention guidelines. In light of the above results, a larger study under a unifying protocol would be warranted.

Mushrooms and soils samples collected from a sub-alpine forest of Mt. Fuji in Japan were measured for 137Cs and stable Cs. The ranges of 137Cs specific activities and stable Cs concentrations in the mushrooms were 291-7950 Bq kg(-1) dry weight and 4.69-58.1 mg kg(-1) dry weight, respectively. Both 137Cs specific activities and stable Cs concentrations in the mushrooms were higher than those in common agricultural plants. The 137Cs specific activities and stable Cs concentrations in the soils were 3.18-149 Bq kg(-1) dry weight and 0.618-2.18 mg kg(-1) dry weight, respectively. The appearance frequencies of filamentous actinomycetes and planktonic bacteria from the soils decreased according to increasing Cs contents in the medium. No relationship was observed between the appearance frequencies of those and the stable Cs concentrations in the soils. The filamentous actinomycetes from any soil sample could not grow in the presence of 25 mM Cs, although the planktonic bacteria from the soil samples could grow with up to 50 mM Cs in YM agar. In addition, the planktonic bacteria from approximately 70% of the soil samples could grow even in the presence of 100 mM Cs. Filamentous actinomycetes were more sensitive to Cs than planktonic bacteria. In in vitro experiments, Cs uptake by these strains of filamentous actinomycetes and planktonic bacteria was high in the presence of 5 mM CsCl and the strains accumulated Cs, the same as in mushrooms. Our results indicate that filamentous actinomycetes in the soils have higher sensitivity to Cs than planktonic bacteria, and several strains of filamentous actinomycetes have a high Cs accumulation in the presence of 5 mM Cs.

Combined treatment of acrinol (Ac) and tetracycline hydrochloride (Tc) against Pseudomonas aeruginosa strains isolated from clinical specimens synergistically increased the bactericidal effect. The minimum bactericidal concentration (MBC) of Ac against P. aeruginosa strain no. 985 was 200 microg/ml, while the MBC of Ac against strains no. 47 and no. 783 was above 800 microg/ml for each. The MBC of Tc was above 400 microg/ml against each of the tested strains. However, simultaneous treatment with 25 microg/ml Ac and 200 microg/ml Tc against P. aeruginosa strain no. 985 decreased the viable cell number from 107 cfu/ml to <10 cfu/ml within 24 h, while a higher concentration of Tc (400 microg/ml) with Ac (25 microg/ml) reduced the viable cell number to <10 cfu/ml within 8 h. A similar synergistic bactericidal effect of Ac and Tc was observed in strains no. 47 and no. 783 by treatment with 200 microg/ml Ac and 200 microg/ml or 400 microg/ml Tc. The degree of bactericidal effect against P. aeruginosa was proportional to the concentration of Tc under the condition of a constant concentration of Ac. Furthermore, Ac-treated cells of strain no. 47 were killed by a following Tc treatment, but cells pretreated with Tc did not show such a sensitivity to Ac. To induce the synergistic effect of Ac and Tc, Ac must be applied to P. aeruginosa before or at the same time as Tc.

A simple and safety gas exposure method was developed using a gas sampling bag as an exposure vessel and a preparation vessel of diluted gas. The gas exposure conditions such as amount of S9 in the plate, volume of gas for the plate, amount of top agar, exposure period and exposure temperature were examined by mutagenicity testing of 1,3-butadiene using the gas sampling bag. Mutagenicity tests of 14 compounds and 1,3-butadiene on S. typhimurium TA98, TA100, TA1535 and TA1537, and E. coli WP2 uvrA were also examined by the developed gas exposure method. 1,3-Butadiene, propyne (methyl acetylene), monochlorodifluoromethane, ethylchloride, diborane and silane were mutagenic. 1-Butene, 2-butene, 2-methylpropene, methyl vinyl ether, trichlorofluoromethane, dichlorodifluoromethane, 1,2-dichloro-1,1,2,2-tetrafluoroethane, 1,1-difluoroethane and phosphine were not mutagenic on S. typhimurium TA98, TA100, TA1535 and TA1537, and E. coli WP2 uvrA with or without metabolic activation. These results were compatible with a previous report, and this developed method has the advantage that it can be tested easily and safely for combustible and self-combustible substances such as 1,3-butadiene and silane.