The Effectiveness of RNAi in Caenorhabditis elegans Is Maintained during Spaceflight

PublishedJournal ArticleResearch Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov'tThis is the final version of the article. Available from Public Library of Science via the DOI in this record.BACKGROUND: Overcoming spaceflight-induced (patho)physiologic adaptations is a major challenge preventing long-term deep space exploration. RNA interference (RNAi) has emerged as a promising therapeutic for combating diseases on Earth; however the efficacy of RNAi in space is currently unknown. METHODS: Caenorhabditis elegans were prepared in liquid media on Earth using standard techniques and treated acutely with RNAi or a vector control upon arrival in Low Earth Orbit. After culturing during 4 and 8 d spaceflight, experiments were stopped by freezing at -80°C until analysis by mRNA and microRNA array chips, microscopy and Western blot on return to Earth. Ground controls (GC) on Earth were simultaneously grown under identical conditions. RESULTS: After 8 d spaceflight, mRNA expression levels of components of the RNAi machinery were not different from that in GC (e.g., Dicer, Argonaute, Piwi; P>0.05). The expression of 228 microRNAs, of the 232 analysed, were also unaffected during 4 and 8 d spaceflight (P>0.05). In spaceflight, RNAi against green fluorescent protein (gfp) reduced chromosomal gfp expression in gonad tissue, which was not different from GC. RNAi against rbx-1 also induced abnormal chromosome segregation in the gonad during spaceflight as on Earth. Finally, culture in RNAi against lysosomal cathepsins prevented degradation of the muscle-specific α-actin protein in both spaceflight and GC conditions. CONCLUSIONS: Treatment with RNAi works as effectively in the space environment as on Earth within multiple tissues, suggesting RNAi may provide an effective tool for combating spaceflight-induced pathologies aboard future long-duration space missions. Furthermore, this is the first demonstration that RNAi can be utilised to block muscle protein degradation, both on Earth and in space.This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, the Japan Society for the Promotion of Science, and “Ground-Based Research Announcement for Space Utilization” promoted by the Japan Space Forum. TE was supported by the Medical Research Council UK (G0801271). NJS was supported by the National Institutes of Health (NIH NIAMS ARO54342). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Hospital and clinic cooperation for the treatment of rheumatoid arthritis in Okayama Prefecture, Japan

Objective: To survey the current status and problems of cooperation between clinics and hospitals in Okayama Prefecture, Japan for the treatment of rheumatoid arthritis (RA). 　Methods: We distributed a questionnaire to 300 of the 983 Okayama Prefecture clinics that had either an internal medicine or orthopedic surgery department, from December 2013 to February 2014. The questionnaire covered practice pattern for RA treatment in clinics, current status of the hospital and clinic cooperation, and acceptance of the biologic therapy. 　Results: One hundred clinics responded to the questionnaire. Seventy percent of the clinics reported making referrals to rheumatologists before the initiation of RA treatment, and half of the other 30% of the clinics administered methotrexate as the first-line treatment for RA by their own decision. Sixty-six clinics cooperated with flagship hospitals, conducting medical and laboratory examinations, providing prescriptions, and treating common diseases of patients. These clinics expected the cooperating rheumatologists to follow-up patients every 3 to 6 months and to make the diagnosis, make decisions regarding RA treatment changes, and perform surgery. Seventy-one percent of the clinics responded that cooperation with a hospital is possible even for patients who are administered biologics. As reasons for no cooperation with the flagship hospitals, clinics noted the lack of information about rheumatologists in the area and recent trends in the management of RA. 　Conclusion: The current study reported, for the first time, the actual conditions of management of RA in clinics, as well as future problems of hospital and clinic cooperation in Okayama Prefecture

STEAL PHENOMENON AFTER LUMBAR AND THORACIC SYMPATHETIC GANGLION BLOCKADE IN DOGS1 THE INFLUENCE OF INTRAVASCULAR VOLUME

The steal phenomenon between the bilateral sides following unilateral sympathetic denervation can be influenced by intravascular blood volume. We studied the changes in skin temperature as an indicator of this phenomenon induced by unilateral lumbar or thoracic sympathetic ganglion blockade in dogs under various hemodynamics. Each of 30 dogs in lumbar and thoracic experiments, which were divided into three groups according to mean right atrial pressure (RAP, mmHg) as hypovolemic (RAP＜3 ; n=10), normovolemic (3≦RAP≦6 ; n=10) and hypervolemic (RAP＞6 ; n=10), underwent'unilat- eral high-frequency thermocoagulation in lumbar (L5-L7) or thoracic (Th7-Th11) sympathetic ganglia. Skin temperature at planta of both hindlimbs or bilateral costal arches was compared among three groups for 60 minutes following･either blockade. Ipsilateral skin temperature rose in all groups (p＜0.05). Contralateral skin temperature fell significantly only in the hypovolemic and the normovolemic groups (p＜0.05), but not in the hypervolemic group. After lumbar sympathetic blockade, the changes is the contralateral skin temperature for 60 minutes, expressed as a percentage of the respective control value, were significantly larger (p＜0.05) in the hypovolemic (-6.4±1.8%) and the normovolemic (-6.0±2.2%) groups than that in the hypervolemic group (+0.1±1.7 %). After thoracic sympathetic blockade, this percentage change was significantly differ- ent (p＜0.05) among the hypovolemic (-1.2±0.7%), the normovolemic (-0.7±0.6%) and the hypervolemic (+0.0±0.4%) groups. We conclude that the steal phenomenon may be influenced by intravascular volume. Therefore, it is necessary to sufficiently understand the patient's systemic hemodynamics or peripheral circulation in performing unilateral sympathetic ganglion blockade. Further- more, a pretreatment such as a preoperative volume loading can be beneficial, especially for the hypovolemic patient who has bilateral vascular disorders

Transforaminal epidural blood patch for intractable spontaneous cerebrospinal fluid leak: a case report

Abstract Background Epidural blood patch (EBP) is a recognized treatment for spontaneous cerebrospinal fluid leak (SCFL) and is typically administered by the interlaminar approach. Here, we report a case of a patient in whom SCFL failed to resolve after three applications of interlaminar EBPs before finally being successfully treated with transforaminal EBP. Case presentation We report a case of a 41-year-old female with a definitive diagnosis of SCFL according to computed tomography (CT) myelography. A fluoroscopy-guided interlaminar EBP was applied three times without resolution of her orthostatic headache. A second myelography was therefore performed demonstrating a leak point on the ventral side of the dura mater. To close the ruptured ventral dura mater, it was necessary to fill the ventral epidural space with blood. Therefore, transforaminal EBP was performed. On spinal CT performed immediately after treatment, the ventral epidural space was observed to be filled with injected blood. Her headache improved the following day, and her symptoms completely subsided after 5\ua0days. Conclusion Transforaminal epidural blood patch is appropriate for patients with intractable cerebrospinal fluid leak. Patients with cerebrospinal fluid leakage due to rupture of the ventral side of the dura mater may be particularly good candidates for this procedure

Kinetically Controlled One-Pot Formation of DEFGH-Rings of Type B Physalins through Domino-Type Transformations

The characteristic DEFGH-ring system of type B physalins has been synthesized by means of a one-pot procedure incorporating domino-type ring transformations. Unexpectedly, we found that introduction of an α-hydroxyester functionality at C17 in ring E allowed the key 7-<i>endo</i> oxy-Michael reaction to proceed. Originally this was thought to be an unfavored process. This afforded the desired caged ring system to be formed in a kinetically controlled manner. Consecutive treatment with AcOH at 100 °C furnished the DEFGH-ring system in one pot