The sialyl-Tn antigen synthase genes regulates migration-proliferation dichotomy in prostate cancer cells under hypoxia.

A low-oxygen (hypoxia) tumor microenvironment can facilitate chemotherapy and radiation therapy resistance in tumors and is associated with a poor prognosis. Hypoxia also affects PCa (prostate cancer) phenotype transformation and causes therapeutic resistance. Although O-glycans are known to be involved in the malignancy of various cancers under hypoxia, the expression and function of O-glycans in PCa are not well understood. In this study, the saccharide primer method was employed to analyze O-glycan expression in PCa cells. Results showed that the expression of sTn antigens was increased in PCa cells under hypoxia. Furthermore, it was found that ST6GalNAc1, the sTn antigen synthase gene, was involved in the migration-proliferation dichotomy and drug resistance in PCa cells under hypoxia. The results of this study will contribute to the development of novel diagnostic markers and drug targets for PCa under hypoxia.

Oxygen isotopes of anhydrous primary minerals show kinship between asteroid Ryugu and comet 81P/Wild2.

The extraterrestrial materials returned from asteroid (162173) Ryugu consist predominantly of low-temperature aqueously formed secondary minerals and are chemically and mineralogically similar to CI (Ivuna-type) carbonaceous chondrites. Here, we show that high-temperature anhydrous primary minerals in Ryugu and CI chondrites exhibit a bimodal distribution of oxygen isotopic compositions: 16O-rich (associated with refractory inclusions) and 16O-poor (associated with chondrules). Both the 16O-rich and 16O-poor minerals probably formed in the inner solar protoplanetary disk and were subsequently transported outward. The abundance ratios of the 16O-rich to 16O-poor minerals in Ryugu and CI chondrites are higher than in other carbonaceous chondrite groups but are similar to that of comet 81P/Wild2, suggesting that Ryugu and CI chondrites accreted in the outer Solar System closer to the accretion region of comets.

Clinical Outcomes of Total Hip Arthroplasty With the Anterolateral Modified Watson-Jones Approach for Displaced Femoral Neck Fractures.

Objective: In recent years, many studies have reported good results with total hip arthroplasty (THA) for displaced femoral neck fractures (FNFs). However, no study has reported the clinical outcomes of the anterolateral modified Watson-Jones THA (MWJ-THA) for displaced FNFs. This study aimed to investigate the clinical results of THA for displaced FNFs at our hospital and to discuss the advantages of MWJ-THA over THA with other approaches for displaced FNFs. Methods: Forty-three patients who underwent MWJ-THA for displaced FNFs were included in this study. Patient characteristics, preinjury walking ability, activities of daily living, implants used, walking ability (at 1, 3, and 6 months after surgery), cup placement angle, clinical hip score, surgical complications, revision surgery, and death within 1 year after surgery were investigated. Results: The mean age of the 43 patients was 63.3 years, and the mean body mass index (kg/m2) was 21.1. Regarding the heads used, 28-mm heads were used in 4 patients, 32-mm heads were used in 32 patients, and 36-mm heads were used in 7 patients. The cups were placed in the Lewinnek safety zone (93.0%). Four patients had stem sinkage of a few millimeters. 6 months postoperatively, 38 patients walked unaided, and 4 patients walked with a cane. The Harris Hip Score averaged over 90 points at all time points. No postoperative dislocation was observed. Two patients died within 1 year postoperatively. Conclusion: In this study, MWJ-THA was performed for displaced FNFs and resulted in no postoperative dislocations. Furthermore, more than 90% of the patients regained their preinjury walking ability at 6 months postoperatively. MWJ-THA has great dislocation control and is effective in treating displaced FNFs.

Functional analysis of GCNT3 for cell migration and EMT of castration-resistant prostate cancer cells.

Castration-resistant prostate cancer (CRPC) is a malignant tumor that is resistant to androgen deprivation therapy. Treatments for CRPC are limited, and no diagnostic markers are currently available. O-glycans are known to play an important role in cell proliferation, migration, invasion, and metastasis of cancer cells. However, the differences in the O-glycan expression profiles for normal prostate cancer (PCa) cells compared with CRPC cells have not yet been investigated. In this study, the saccharide primer method was employed to analyze the O-glycans expressed in CRPC cells. Expression levels of core 4-type O-glycans were significantly increased in CRPC cells. Furthermore, the expression level of N-Acetylglucosaminyltransferase 3 (GCNT3), a core 4-type O-glycan synthase gene, was increased in CRPC cells. The expression of core 4-type O-glycans and GCNT3 was presumed to be regulated by androgen deprivation. GCNT3 knockdown induced cell migration and epithelial-mesenchymal transition. These observations elucidate the mechanism of acquisition of castration resistance in PCa and offer new possibilities for the development of diagnostic markers and therapeutic targets in the treatment of PCa.

On the origin and evolution of the asteroid Ryugu: A comprehensive geochemical perspective.

Presented here are the observations and interpretations from a comprehensive analysis of 16 representative particles returned from the C-type asteroid Ryugu by the Hayabusa2 mission. On average Ryugu particles consist of 50% phyllosilicate matrix, 41% porosity and 9% minor phases, including organic matter. The abundances of 70 elements from the particles are in close agreement with those of CI chondrites. Bulk Ryugu particles show higher δ18O, Δ17O, and ε54Cr values than CI chondrites. As such, Ryugu sampled the most primitive and least-thermally processed protosolar nebula reservoirs. Such a finding is consistent with multi-scale H-C-N isotopic compositions that are compatible with an origin for Ryugu organic matter within both the protosolar nebula and the interstellar medium. The analytical data obtained here, suggests that complex soluble organic matter formed during aqueous alteration on the Ryugu progenitor planetesimal (several 10's of km), <2.6 Myr after CAI formation. Subsequently, the Ryugu progenitor planetesimal was fragmented and evolved into the current asteroid Ryugu through sublimation.

In vitro synthesis of mucin-type O-glycans using saccharide primers comprising GalNAc-Ser and GalNAc-Thr residues.

Mucin-type O-glycosylation of serine or threonine residue in proteins is known to be one of the major post-translational modifications. In this study, two novel alkyl glycosides, Nα-lauryl-O-(2-acetamido-2-deoxy-α-d-galactopyranosyl)-l-serineamide (GalNAc-Ser-C12) and Nα-lauryl-O-(2-acetamido-2-deoxy-α-d-galactopyranosyl)-l-threonineamide (GalNAc-Thr-C12) were synthesized as saccharide primers to prime mucin-type O-glycan biosynthesis in cells. Upon incubating human gastric cancer MKN45 cells with the saccharide primers, 22 glycosylated products were obtained, and their structures were analyzed using liquid chromatography-mass spectrometry and enzyme digestion. The amounts of glycosylated products were dependent on the amino acid residues in the saccharide primers. For example, in vitro synthesis of T antigen (Galß1-3GalNAc), fucosyl-T (Fucα1-2Galß1-3GalNAc), and sialyl-T (NeuAcα2-3Galß1-3GalNAc) preferred a serine residue, whereas sialyl-Tn (NeuAcα2-6GalNAc) preferred a threonine residue. Furthermore, the glycosylated products derived from GalNAc-Ser/Thr-C12 and Gal-GalNAc-Ser/Thr-C12 using cell-free synthesis showed the same amino acid selectivity as those in the cell experiments. These results indicate that glycosyltransferases involved in the biosynthesis of mucin-type O-glycans distinguish amino acid residues conjugated to GalNAc. The saccharide primers developed in this study might be useful for comparing mucin-type oligosaccharides in cells and constructing oligosaccharide libraries to study cell function.

Construction of an infectious dahlia common mosaic virus clone.

Dahlia is a major ornamental plant that is cultivated worldwide. However, dahlia plants, which are mainly propagated through vegetative reproduction, are susceptible to widespread damage by viruses, and viral control requires that the nature of the infecting virus(es) be known. In this study, dahlia common mosaic virus (DCMV) was detected for the first time in Japan and sequenced. This is the first report of an infectious DCMV clone being constructed, and it will aid in the characterization of DCMV.

Development of a Spontaneous Emulsifying Technique Using Porous Microparticles with Water-soluble Polymers.

A novel spontaneous emulsification method using porous polymer particles was investigated for the facile preparation of emulsions without mechanical manipulation. Porous water-soluble polymer particles prepared by spray freeze-drying could absorb soybean oil via capillary action. When the particles were added to water, emulsification proceeded rapidly with the dissolution of the polymer. The importance of using a water-soluble polymer for particle formation for the formation of fine emulsions and maintenance of dispersibility was confirmed. This emulsification technology is expected to be applied to the development of formulations that improve the solubility and mucosal absorption of poorly water-soluble drugs.

A Lipid Bilayer Formed on a Hydrogel Bead for Single Ion Channel Recordings.

Ion channel proteins play important roles in various cell functions, making them attractive drug targets. Artificial lipid bilayer recording is a technique used to measure the ion transport activities of channel proteins with high sensitivity and accuracy. However, the measurement efficiency is low. In order to improve the efficiency, we developed a method that allows us to form bilayers on a hydrogel bead and record channel currents promptly. We tested our system by measuring the activities of various types of channels, including gramicidin, alamethicin, α-hemolysin, a voltage-dependent anion channel 1 (VDAC1), a voltage- and calcium-activated large conductance potassium channel (BK channel), and a potassium channel from Streptomyces lividans (KcsA channel). We confirmed the ability for enhanced measurement efficiency and measurement system miniaturizion.

Preventive Effect of Alginate Gel Formulation on Perineural Adhesion.

Background: Perineural adhesion is a potential complication of manipulating peripheral nerves. Using a model of median nerve manipulation in the carpal tunnel, perineural adhesion preventive effects of an alginate gel formulation were examined. Methods: After exposing carpal tunnels of Japanese white rabbits and dissecting the median nerve, the gliding floor was excised as much as possible and the transverse carpal ligament was repaired to induce a perineural tissue reaction. Prior to wound closure, 0.5 ml of alginate gel formulation was administered into the right carpal tunnel (formulation group) and 0.5 ml of physiological saline was administered into the left carpal tunnel (control group). At 1, 2, 3, and 6 weeks after treatment, electrophysiological evaluation of thenar distal latency, macroscopic evaluation with adhesion score, and pathological evaluation of carpal tunnel cross sections were performed (N = 4-5 at each time point). Results: Although distal latency tended to be low in the formulation group, there was no significant difference between the groups according to electrophysiological evaluation. Macroscopic evaluation revealed that the adhesion score was always lower in the formulation group than in the control group; over the course of treatment, it remained unchanged in the formulation group, but peaked at 3 weeks after treatment in the control group. In pathological evaluation, neural perfusion peaked at 2-3 weeks after treatment in both groups; neural perfusion tended to be lower in the formulation group than in the control group. Conclusions: Results suggested that the peak tissue response associated with nerve dissection occurred 2-3 weeks after treatment and that the repair process started subsequently. The alginate gel formulation modified the surrounding environment of the nerve and promoted repair by acting as a physical barrier against perineural fibrosis. The preventive effect of alginate gel on perineural adhesion may improve treatment outcomes of constrictive neuropathy.

Differentiated adipose-derived stem cells promote peripheral nerve regeneration.

INTRODUCTION: Many reports have indicated that adipose-derived stem cells (ADSCs) are effective for nerve regeneration. We investigated nerve regeneration by combining a polyglycolic acid collagen (PGA-c) tube, which is approved for clinical use, and Schwann cell-like differentiated ADSCs (dADSCs). METHODS: Fifteen-millimeter-long gaps in the sciatic nerve of rats were bridged in each group using tubes (group I), with tubes injected with dADSCs (group II), or by resected nerve (group III). RESULTS: Axonal outgrowth was greater in group II than in group I. Tibialis anterior muscle weight revealed recovery only in group III. Latency in nerve conduction studies was equivalent in group II and III, but action potential was lower in group II. Transplanted dADSCs maintained Schwann cell marker expression. ATF3 expression level in the dorsal root ganglia was equivalent in groups II and III. DISCUSSION: dADSCs maintained their differentiated state in the tubes and are believed to have contributed to nerve regeneration.

Persistent colonization of non-lymphoid tissue-resident macrophages by Stenotrophomonas maltophilia.

Accumulating evidence has revealed that lymphoid tissue-resident commensal bacteria (e.g. Alcaligenes spp.) survive within dendritic cells. We extended our previous study by investigating microbes that persistently colonize colonic macrophages. 16S rRNA-based metagenome analysis using DNA purified from murine colonic macrophages revealed the presence of Stenotrophomonas maltophilia. The in situ intracellular colonization by S. maltophilia was recapitulated in vitro by using bone marrow-derived macrophages (BMDMs). Co-culture of BMDMs with clinically isolated S. maltophilia led to increased mitochondrial respiration and robust IL-10 production. We further identified a 25-kDa protein encoded by the gene assigned as smlt2713 (recently renamed as SMLT_RS12935) and secreted by S. maltophilia as the factor responsible for enhanced IL-10 production by BMDMs. IL-10 production is critical for maintenance of the symbiotic condition, because intracellular colonization by S. maltophilia was impaired in IL-10-deficient BMDMs, and smlt2713-deficient S. maltophilia failed to persistently colonize IL-10-competent BMDMs. These findings indicate a novel commensal network between colonic macrophages and S. maltophilia that is mediated by IL-10 and smlt2713.

ZF-Mapper: Simple and Complete Freeware for Fluorescence Quantification in Zebrafish Images.

Zebrafish embryos and larvae have become popular vertebrate models because their body walls are transparent, which enables live imaging of target organs using fluorescent protein transgenes or dye staining. Software packages for the quantification of these fluorescent signals are available from both commercial and noncommercial sources; however, their algorithms are complicated and their resources (code) have mostly not been openly shared. In this study, we developed a simple and robust open-source software tool named "ZF-Mapper" for the quantification of the fluorescence intensity of each pixel in zebrafish images with batch image file processing capability. Using this software, we can evaluate the three-dimensional (3D) distribution of fluorescence intensity among zebrafish cells by analyzing each image pixel. We tested ZF-Mapper for the analysis of zebrafish with macrophage-specific enhanced green fluorescent protein (EGFP) and obtained results that were equivalent to those acquired using the conventional image analysis software ImageJ. We further applied ZF-Mapper to the analysis of zebrafish with cancer cell xenografts and quantified the amount of implanted melanoma cells labeled with a tdTomato red fluorescent protein in the whole body and the tail region. In addition, by combining ZF-Mapper with R freeware, we created an interactive 3D scatter plot of the fluorescence intensities of macrophage-EGFPs in zebrafish. In summary, we developed the Python-based freeware ZF-Mapper for the quantification of fluorescent signals in multiple zebrafish images, which enables fluorescence-based zebrafish screening. We provide the source code and the executable application software for Windows (.exe) and macOS (.app).

Tin Ring Splint Treatment for Osteoarthritis of the Distal Interphalangeal Joints.

Background: We made a tin ring splint for osteoarthritis of the distal interphalangeal joint that looks attractive and is easy to wear. We report the treatment results with this splint. Methods: We enrolled 30 patients with painful osteoarthritis of the distal interphalangeal joint in this study. A tin ring splint was made with tin alloy containing small quantities of silver. Patients were instructed to wear the splint when they felt pain. Patients were assessed before splint use and after 1, 3, and 6 months of splint use. Endpoints included the numeric pain scale, active arc of motion of the distal interphalangeal joint, Hand 20, functional assessment criteria of the upper extremities, and treatment satisfaction. In addition, data were collected on time to symptom relief and satisfaction related to usability and appearance of the splint (0 = dissatisfied, 10 = satisfied). Results: The numeric pain scale showed significant pain improvement from 58.4 ± 4.1 at baseline to 33.1 ± 4.5 at 1 month, and the Hand 20 score also showed significant improvement from 35.0 ± 4.3 at baseline to 20.2 ± 3.2 after 6 months. Active arc of motion were not changed significantly. Most patients responded that symptoms were relieved by the 10th day after treatment. Satisfaction related to usability was 8.9 ± 0.3, and appearance was 7.6 ± 0.4. Conclusions: A tin ring splint quickly reduced pain, and satisfaction related to usability and appearance was high. This splint could be one choice for conservative treatment of osteoarthritis of the distal interphalangeal joint.

Decreased aluminium tolerance in the growth of Saccharomyces cerevisiae with SSO2 gene disruption.

Aluminium ions inhibit growth of the budding yeast Saccharomyces cerevisiae. Disruption of the SSO2 gene increased the susceptibility to aluminium. Sso2p belongs to the soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) family. SSO2 has one paralogue, SSO1, which encodes Sso1p. The SNARE complex containing Sso1/2p plays a role in the recognition of plasma membrane targeted vesicle transport. The susceptibility to aluminium stress was not increased in the Δsso1 strain. The phenotype of aluminium ion influx between the wild-type and Δsso2 strains was not different, suggesting that Sso2p was involved in the elimination of cellular aluminium. However, the cellular lipid constitution of Δsso2 was richer in unsaturated fatty acids than the wild type, indicating that Sso2p is associated with lipid homeostasis of the plasma membrane. Aluminium treatment increased the production of reactive oxygen species (ROS) during proliferation. ROS production was increased in the Δsso2 strain after 3 h of aluminium treatment compared with the wild type. These results suggested that Sso2p plays a role in maintaining the lipid composition of the plasma membrane and the increase in unsaturated fatty acids amplified the production of ROS in the acute phase of aluminium stress. ROS derived from aluminium stress inhibited growth and resulted in the susceptibility of the Δsso2 strain.

Non-tuberculous Mycobacterium or Fungus Induced Chronic Tenosynovitis with Rice Body of the Hand.

BACKGROUND: Chronic tenosynovitis of the wrist and hand is commonly seen by orthopedists, especially hand surgeons. However, cases with rice body formation are comparatively rare. Thus, we retrospectively reviewed the cases of chronic tenosynovitis in our department and evaluated the necessity of antibiotic therapy in the early post-surgical stage. METHODS: We analyzed the medical and surgical records of patients who had undergone surgery for chronic tenosynovitis with rice body formation in our department from 1997 to 2015. We evaluated the causes of chronic tenosynovitis, culture findings, pathological findings, and post-operative treatment courses. RESULTS: Nineteen patients with 23 involved hands underwent surgery for chronic tenosynovitis, and 9 patients had rice body formation. The most common cause of chronic tenosynovitis was non-tuberculous mycobacteriosis, and other causes were fungal infection and infection of unknown origin. Recurrence was observed in 2 cases of mycobacteriosis and 1 case of fungal infection; 1 case of mycobacteriosis also had a re-recurrence. CONCLUSIONS: In the diagnosis of chronic tenosynovitis with rice body formation, it is necessary to consider not only non-tuberculous mycobacteriosis, but also fungal infection as its origin. However, it is difficult to define the cause of synovitis, but in cases in which these infections are suspected, anti-bacterial therapy in the early post-surgical period could be effective.

Computational design of a symmetrical ß-trefoil lectin with cancer cell binding activity.

Computational protein design has advanced very rapidly over the last decade, but there remain few examples of artificial proteins with direct medical applications. This study describes a new artificial ß-trefoil lectin that recognises Burkitt's lymphoma cells, and which was designed with the intention of finding a basis for novel cancer treatments or diagnostics. The new protein, called "Mitsuba", is based on the structure of the natural shellfish lectin MytiLec-1, a member of a small lectin family that uses unique sequence motifs to bind α-D-galactose. The three subdomains of MytiLec-1 each carry one galactose binding site, and the 149-residue protein forms a tight dimer in solution. Mitsuba (meaning "three-leaf" in Japanese) was created by symmetry constraining the structure of a MytiLec-1 subunit, resulting in a 150-residue sequence that contains three identical tandem repeats. Mitsuba-1 was expressed and crystallised to confirm the X-ray structure matches the predicted model. Mitsuba-1 recognises cancer cells that express globotriose (Galα(1,4)Galß(1,4)Glc) on the surface, but the cytotoxicity is abolished.

Galeazzi-equivalent Fractures: Report of Two Cases and Literature Review.

The Galeazzi-equivalent fracture is a rare injury that occurs in children. The most important issue is the distal ulnar epiphyseal injury. Although there have been some case reports, most of them performed only short term follow-up. This article describes two cases of this fracture with long term follow-up until epiphyseal closure. First case is a 12-year-old girl who sustained a Galeazziequivalent fracture of her right forearm and underwent emergency surgery. At follow-up of 5 years and 10 months postsurgery, radiographs show ulnar growth arrest of one mm and she has mild pain. Second case is a 15-year-old boy who sustained an open Galeazzi-equivalent fracture of his left forearm and underwent emergency surgery. At follow-up of 3 years and 3 months postsurgery, radiographs show no growth arrest of the distal ulna. He has no residual complaint. Long term follow-up is absolutely necessary to monitor ulnar growth.