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.

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.

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.

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.

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.

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.

Catfish rhamnose-binding lectin induces G0/1 cell cycle arrest in Burkitt's lymphoma cells via membrane surface Gb3.

Silurus asotus egg lectin (SAL), an α-galactoside-binding protein isolated from the eggs of catfish, is a member of the rhamnose-binding lectin family that binds to Gb3 glycan (Galα1-4Galß1-4Glc). We have previously demonstrated that SAL reduces the proliferation of Gb3-expressing Burkitt's lymphoma Raji cells and confirm here that it does not reduce their viability, indicating that unlike other lectins, it is not cytotoxic. The aim of this study was to determine the signal transduction mechanism(s) underlying this novel SAL/Gb3 binding-mediated effect profile. SAL/Gb3 interaction arrested the cell cycle through increasing the G0/1 phase population of Raji cells. SAL suppressed the transcription of cell cycle-related factors such as c-MYC, cyclin D3, and cyclin-dependent protein kinase (CDK)-4. Conversely, the CDK inhibitors p21 and p27 were elevated by treatment with SAL. In particular, the production of p27 in response to SAL treatment increased steadily, whereas p21 production was maximal at 12 h and lower at 24 h. Activation of Ras-MEK-ERK pathway led to an increase in expression of p21. Notably, treatment of Raji cells with anti-Gb3 mAb alone did not produce the above effects. Taken together, our findings suggest that Gb3 on the Raji cell surface interacts with SAL to trigger sequential GDP-Ras phosphorylation, Ras-MEK-ERK pathway activation, p21 production, and cell cycle arrest at the G0/1 phase.

cDNA and Gene Structure of MytiLec-1, A Bacteriostatic R-Type Lectin from the Mediterranean Mussel (Mytilus galloprovincialis).

MytiLec is an α-d-galactose-binding lectin with a unique primary structure isolated from the Mediterranean mussel (Mytilus galloprovincialis). The lectin adopts a ß-trefoil fold that is also found in the B-sub-unit of ricin and other ricin-type (R-type) lectins. We are introducing MytiLec(-1) and its two variants (MytiLec-2 and -3), which both possess an additional pore-forming aerolysin-like domain, as members of a novel multi-genic "mytilectin family" in bivalve mollusks. Based on the full length mRNA sequence (911 bps), it was possible to elucidate the coding sequence of MytiLec-1, which displays an extended open reading frame (ORF) at the 5' end of the sequence, confirmed both at the mRNA and at the genomic DNA sequence level. While this extension could potentially produce a polypeptide significantly longer than previously reported, this has not been confirmed yet at the protein level. MytiLec-1 was revealed to be encoded by a gene consisting of two exons and a single intron. The first exon comprised the 5'UTR and the initial ATG codon and it was possible to detect a putative promoter region immediately ahead of the transcription start site in the MytiLec-1 genomic locus. The remaining part of the MytiLec-1 coding sequence (including the three sub-domains, the 3'UTR and the poly-A signal) was included in the second exon. The bacteriostatic activity of MytiLec-1 was determined by the agglutination of both Gram-positive and Gram-negative bacteria, which was reversed by the co-presence of α-galactoside. Altogether, these data support the classification of MytiLec-1 as a member of the novel mytilectin family and suggest that this lectin may play an important role as a pattern recognition receptor in the innate immunity of mussels.

A pediatric case of single-level idiopathic cervical intervertebral disk calcification with symptom relapse 1 year after initial onset.

BACKGROUND: To date there has been only 1 reported case of the symptom relapse of pediatric idiopathic intervertebral disk calcification (PIIDC), as described by Yoon and colleagues in 1987, who reported symptom relapse in a patient with multilevel PIIDC. Thus, symptom relapse in patients with single-level PIIDC have not been reported. METHODS: We report here a case of single-level PIIDC with symptom relapse 1 year after the initial onset. RESULTS: The patient was a 7-year-old girl who developed cervical pain and fever up to 38°C without an obvious cause. Computed tomography (CT) revealed calcification in the C4/5 intervertebral disk space and in the epidural space at the C3-5 vertebral levels. The patient was diagnosed with PIIDC and treatment with oral nonsteroidal anti-inflammatory drugs was begun. Both cervical pain and fever gradually improved and resolved in approximately 1 week. CT obtained 6 months after the initial onset showed calcifications localized in the posterior area of the C4/5 intervertebral disk space and reduced epidural calcifications, which had nearly resolved. One year after the initial onset, the patient developed similar symptoms. CT revealed an enlarged calcified lesion in the epidural space. Thus, the patient was diagnosed with symptom relapse of PIIDC. Although there was enlargement of calcifications in the epidural space, there were no calcifications involving the intervertebral disk at the time of relapse. The patient was treated conservatively. Follow-up CT revealed that the lesion resolved with time. CONCLUSIONS: This report described a patient with single-level PIIDC and symptom relapse 1 year after the initial onset. In the case presented herein, calcifications of the intervertebral space had extruded into the epidural space, thus causing a symptom relapse. The patient was treated conservatively at the initial onset and at the time of relapse. The symptoms improved both times. Although patients with single-level PIIDC usually have an uneventful clinical course, it is necessary to be mindful of potential symptom relapse.

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.

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.

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.

Balanced polarization maintaining fiber Sagnac interferometer vibration sensor.

To achieve a nearly zero-delay operating point in a polarization-maintaining (PM) fiber Sagnac interferometer, two identical PM fibers were incorporated so that their two main axes were orthogonally coupled to each other. A simple fiber vibration sensor system was constructed with a light emitting diode and a balanced PM fiber Sagnac interferometer, in which one of the PM fibers was used as a sensing cable and the other as a reference cable. The vibration sensor was confirmed to be temperature-compensated and generated a phase shift per unit length and unit strain of the sensor of 4.7 milliradian/(m·µÎµ) when mechanical vibrations with 1 kHz sinusoidal and triangular waves were stably observed under an input power of 10 µW.

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.

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).