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CEBPA-IGH, a fusion gene of the immunoglobulin heavy-chain locus (IGH) and the CCAAT enhancer-binding protein α (C/EBPα) gene, is recurrently found in B-ALL cases and causes aberrant expression of C/EBPα, a master regulator of granulocyte differentiation, in B cells. Forced expression of C/EBPα in B cells was reported to cause loss of B-cell identity due to the inhibition of Pax5, a master regulator of B-cell differentiation; however, it is not known whether the same mechanism is applicable for B-ALL development by CEBPA-IGH. It is known that a full-length isoform of C/EBPα, p42, promotes myeloid differentiation, whereas its N-terminal truncated isoform, p30, inhibits myeloid differentiation through the inhibition of p42; however, the differential role between p42 and p30 in ALL development has not been clarified. In the present study, we examined the effect of the expression of p42 and p30 in B cells by performing RNA-seq of mRNA from LCL stably transfected with p42 or p30. Unexpectedly, suppression of PAX5 target genes was barely observed. Instead, both isoforms suppressed the target genes of MEF2 family members (MEF2s), other regulators of B-cell differentiation. Similarly, MEF2s target genes rather than PAX5 target genes were suppressed in CEBP-IGH-positive ALL (n = 8) compared with other B-ALL (n = 315). Furthermore, binding of both isoforms to MEF2s target genes and the reduction of surrounding histone acetylation were observed in ChIP-qPCR. Our data suggest that the inhibition of MEF2s by C/EBPα plays a role in the development of CEBPA-IGH-positive ALL and that both isoforms work co-operatively to achieve it.

・In Arabidopsis thaliana, PROPEPs and their derived elicitor‐active Pep epitopes provide damage‐associated molecular patterns (DAMPs), which trigger defence responses through cell‐surface receptors PEPR1 and PEPR2. In addition, Pep peptides induce root growth inhibition and root hair formation, however their relationships and coordinating mechanisms are poorly understood. ・Here, we reveal that Pep1‐mediated root hair formation requires PEPR‐associated kinases BAK1/BKK1 and BIK1/PBL1, ethylene, auxin and root hair differentiation regulators, in addition to PEPR2. ・Our analysis on 69 accessions unravels intraspecies variations in Pep1‐induced root hair formation and growth inhibition. The absence of a positive correlation between the two traits suggests their separate regulation and diversification in natural populations of A. thaliana. ・Restricted PEPR2 expression to certain root tissues is sufficient to induce root hair formation and growth inhibition in response to Pep1, indicating the capacity of non‐cell‐autonomous receptor signalling in different root tissues. Of particular note, root hair cell‐specific PEPR2 expression uncouples defence activation from root growth inhibition and root hair formation, suggesting a unique property of root hairs in root defence activation following Pep1 recognition.

In the drug development process, it is important to assess the contributions of drug-metabolizing enzymes and/or drug transporters to the intestinal pharmacokinetics of candidate compounds. For such assessments, chemical inhibitors are often used in in vitro systems. However, this practice poses two problems: one is the low expression levels of pharmacokinetic-related genes in conventional in vitro systems, such as Caco-2 cells, and the other is the off-target and less-efficient effects of their inhibitors. Here, as a model, we have established human biopsy-derived enteroids deficient in MDR1, a key efflux transporter. The expression levels and activities of other pharmacokinetic-related genes, such as CYP3A4, in the MDR1-knockout (KO) enteroid-derived monolayers were maintained at levels as high as those in the WT enteroid-derived monolayers. The contribution of MDR1 to the cytotoxicity of vinblastine, which CYP3A4 metabolized, was accurately evaluated by using the MDR1-KO enteroid-derived monolayers. In contrast, it could not be evaluated in the WT enteroid-derived monolayers treated by verapamil, a widely used MDR1 inhibitor, due to the off-target effect of verapamil, which also inhibits CYP3A4. The combination of human enteroid-derived monolayers and genome editing technology would be a powerful tool to evaluate the contributions of specific pharmacokinetic-related molecules.

Protein S is a vitamin K-dependent glycoprotein with important anticoagulant, fibrinolytic, anti-inflammatory, anti-apoptotic, and cytoprotective functions. Congenital protein S deficiency is an autosomal dominant thrombophilia due to protein S gene (PROS1) variations. Our group identified a variation in PROS1 that translates into protein S deficiency: c.50 T C (p.Leu17Pro). Here, we investigated the mechanisms by which this variation results in protein S deficiency.The effect of L17P substitution on protein S signal peptide was predicted by in silico (a computational prediction technique) analysis of hydrophobicity and signal peptide cleavage. Recombinant protein S was overexpressed in HEK293 and COS-7 cells. Intracellular kinetics and extracellular secretion of recombinant protein S-L17P were analyzed by western blotting and immunocytochemistry.In silico hydrophobicity analysis showed that protein S-L17P had disrupted hydrophobic status in the h-region of its signal peptide. Under normal culture conditions, recombinant protein S -L17P was not detected in either transfectant cell lysates or medium. Upon treatment with a proteasome inhibitor, recombinant protein S-L17P was clearly detected in the cell lysate, but not in the culture medium. Recombinant protein S-L17P did not undergo post-translational modification with N-glycosylation, suggesting that the nascent polypeptide of recombinant protein S-L17P is not transported to the endoplasmic reticulum lumen, but is mislocalized to the cytosol.PROS1-L17P variation translates into protein S deficiency. Protein S-L17P causes its cytosolic mislocalization resulting in its proteasome-dependent degradation.

Recognition of microbe- and danger-associated molecular patterns (MAMPs and DAMPs, respectively) by pattern recognition receptors (PRRs) is central to innate immunity in both plants and animals. The plant PRRs described to date are all cell surface-localized receptors. According to their ligand-binding ectodomains, each PRR engages a specific coreceptor or adaptor kinase in its signaling complexes to regulate defense signaling. With a focus on the coreceptor RLK BRI1-ASSOCIATED RECEPTOR KINASE1 (BAK1) and related SOMATIC EMBRYOGENESIS RECEPTOR KINASEs (SERKs), here we review the increasing inventory of BAK1 partners and their functions in plant immunity. We also discuss the significance of autoimmunity triggered by BAK1/SERK4 disintegration in shaping the strategies for attenuation of PRR signaling by infectious microbes and host plants.

Background Medial meniscal extrusion (ME) is a biomarker to predict later development of knee osteoarthritis (KOA). On the other hand, we have reported osteophyte formation at the posterior condylar notch of the femur served as a biomarker for the same purpose. The purpose of this study is to compare capacity of the two biomarkers in predicting KOA development. Methods Two cohort of knees were established utilizing publicly available data from the Osteoarthritis Initiative (OAI). No OA group (NOA) consisted of knees that were grade 0 or 1 on Kellgren and Lawrence grade (K/L) both at baseline and 48 months later, and pre-radiographic-OA group (PROA) consisted of knees that were grade 0 or 1 at baseline but grade ≥2 48 months later. Baseline MR images were evaluated in terms of ME and osteophyte formation at the posterior condylar notch. ME was evaluated both by meniscus subluxation index (MSI) indicating the ratio of the extruded width of the medial meniscus to the width of medial meniscal body and by the medial radial displacement (MRD) indicating actual extruded width. The size of the osteophyte was assessed using a semi-quantitative whole-organ magnetic resonance imaging score (WORMS). The predictive accuracy of KOA was assessed by the area under the receiver operating characteristic curve (AUC) and optimal cutoff was determined for each parameter. Results The AUC for MSI was 0.654 (0.561–0.748: 95% CI) and the cutoff value was determined as 17%. That for MRD was 0.677 (0.584–0.770) and the cutoff value was 2.2 mm. The AUC for the WORMS score at the posterior condylar notch was 0.667 (0.579–0.756) and the cutoff value was 2. Conclusions Similar predicting capacity of KOA development was found both in ME and osteophyte formation at the posterior condylar notch. Using these simple parameter, mas-screening for KOA development would be possible.

Study Design: Prospective study. Purpose: To examine the long-term effects of interspinous ligament injections of local anesthetics and steroids for the treatment of Baastrup’s diseases. Overview of Literature: Baastrup’s disease is associated with axial low back pains. Baastrup’s disease has been more recently described as the “kissing spinous processes” disease. Several authors have reported methods for the diagnosis and treatment of the disease. However, there has been only one report of patients receiving interspinous ligament injections of agents for the treatment of Baastrup’s disease. Methods: Seventeen patients showed severe low back pains between spinous processes at L3–L4 or L4–L5. X-ray imaging, computed tomography, and magnetic resonance imaging revealed kissing spinous processes, consolidation of spinous process, or inflamma tion of an interspinous ligament. Pain reliefs after lidocaine and dexamethasone administration into interspinous ligament as therapy for low back pains were being examined and followed up. Results: Low back pain scores significantly improved immediately after injection of the agents into interspinous ligaments. At final follow-up (1.4 year), low back pain scores significantly improved as compared with before the treatment. Conclusions: Findings from the current study indicate that lidocaine and dexamethasone administration into interspinous ligament in patients diagnosed with Baastrup’s disease is effective for managing the pain associated with this disease.

A first assessment of anammox activity, which is a unique N2 emission process, was conducted in samples of coastal marine sediment from Japan with a 15N tracer. The occurrence and diversity of bacteria possibly responsible for the anammox process were also evaluated by selective PCR-amplification of the 16S rRNA gene for known anammox bacteria. Anammox activity, detected by measuring 14N15N gas production, was only found in samples collected at the intertidal sand bank located at the Yodo River estuary. In the Yodo River samples, 16S rRNA gene fragments affiliated with the known anammox bacterial lineage were also recovered, and the two major phylotypes were both "Candidatus Scalindua wagneri" relatives with 95% and 98% sequence similarity. Even from the other samples in which no recognizable anammox activity was detected, 16S rRNA gene fragments related to known anammox bacteria, but not to "Ca. S. wagneri", were detected. This is the first report of anammox-mediated N2 emission in coastal marine environments in Japan. Notably, the PCR-based analysis allowed us to discover unexpected phylogenetic diversity of anammox bacteria-related 16S rRNA gene sequences. The selective PCR primer set developed in this study could be a powerful tool to unveil the ecology of anammox bacteria in natural environments.

Although the emission of N(2) via anaerobic ammonium oxidation (anammox) is a key process in the elimination of nitrogenous compounds from aquatic environments, little information is available regarding its significance and the relevant microorganisms (anammox bacteria) in eutrophic freshwater lakes. In the present study, the anammox bacteria in the sediment of a eutrophic lake in Japan, Lake Kitaura, were examined using a (15)N-tracer technique to measure their potential anammox activity. Potential anammox activity was localized to the northern region of the lake where a stable supply of both NH(4)(+) and NO(3)(-) existed in the sediment. These results suggest the contribution of anammox bacteria to the total emission of N(2) from sediment in this eutrophic lake to not be negligible. Moreover, selective PCR successfully amplified anammox bacteria-related (Brocadiales-related) 16S rRNA genes from sediment samples in which potential anammox activity was observed. The clone libraries consisted of diverse phylotypes except the genus "Scalindua"-lineages, and the lineages of genus "Brocadia" were dominantly recovered, followed by the genus "Kuenenia"-lineages. Most of them, however, were novel and phylogenetically distinguishable from known Brocadiales species. A unique population of anammox bacteria inhabits and potentially contributes to the emission of N(2) from Lake Kitaura.

Combined molecular mechanics (MM) and semiempirical molecular-orbital (MO) calculations have been applied to the investigation on the conformational and electronic properties of periplanone-B (1), a major component of the sex pheromone of American cockroach, and structurally related analogues 2, 3, 4 and 5. In the first step, the geometries of conformers of 1–5 were obtained by MM2 combined with a new algorithm for exhaustive generation of ring conformations, CONFLEX2, and an additional set of MM2 force-field parameters. The global minimum (1A) of the natural pheromone populates predominantly and is superimposable on the X-ray structure. Minor conformers 1B and 1C correspond to the rotamers at the isopropyl group of 1A. The global minimum of the analogue 2 was identified to have the structure 2A, which agrees well with the X-ray data. The structural comparison of the stable conformers of the analogues 2–5 with 1A revealed similar ring conformations in the most stable conformers of 2, 3 and 4, and the third one of 5. These results suggest that the ring structure characteristic of the conformer 1A and the conformer populations must be a significant factor in the biological activity of these analogues. In the next step, the electronic properties were calculated by the semiempirical MO (AM1) method. A significant correlation is found between the biological activity and one of the unoccupied frontier orbitals which is localized around C1–C10–C9 including the carbonyl and spiro-epoxy groups. A newly defined effective frontier parameter: EF(N)(s), which regards the orbital electron density, orbital energy, and ring conformation is found to correlate well with the biological activity of these analogues.

Electroantennogram and single sensillum recordings were performed on the male cigarette beetle, Lasioderma serricorne , antennae. Strong antennal responses were elicited by not only natural sex pheromone component (4S,5S,7S)-serricornin (SSS-serricornin) but also non-natural behavioural inhibitor, (4S,5S,7R)-serricornin (SSR-serricornin). In the case of mixture application of the SSS- and SSR-serricornin, integrated electroantennogram responses were observed. Single sensillum recording from male flagella sensilla showed that the majority of basiconic type sensilla responded to the behavioural inhibitor as well as to the sex pheromone. However, a few of them responded only to the inhibitor. The behavioural inhibitory action of SSR-serricornin toward the SSS-serricornin results from central integration of the information from separate receptor cells rather than blockage at the peripheral receptors.

The biological activities of synthetic periplanone analogs, including four candidates of periplanone-A (P-A), were evaluated by behavioral and electroantennogram (EAG) assays. Among 16 periplanone analogs, six compounds evoked pheromonal activity from the male American cockroaches. The threshold dosages of these biological active analogs were 10-10(5) times lower than that of the known periplanone mimic, germacrene-D. The conformation required for eliciting the pheromonal activity is discussed in terms of the structure-activity relationship of these analogs. Hauptmann's P-A elicited the strongest pheromonal activity among four candidates of P-A in our bioassay, suggesting that Hauptmann's P-A is a natural P-A produced from female cockroaches.

Conformational analysis using molecular mechanics (MM) was performed for a determination of the stereochemistry of serricornin, the sex pheromone of the cigarette beetle (Lasioderma serricorne F.). An exhaustive conformational analysis using MM2 calculations with algorithms for covering torsional energy surfaces of flexible molecules furnishes coordinates and steric energies of all local energy minimum conformers of serricornin, both acyclic and the corresponding cyclic forms. These coordinates gave angles required for the calculation of vicinal H/H coupling constants ((3) J HHs) of each energy minima by Altona's modified Karplus equation. The Boltzmann distributions of all local energy minima were calculated from their steric energies to furnish populations of each energy minimum conformer. Populationweighted averaged(3) J HHs of four enantiomeric pairs, (S (*),S (*),S (*))-, (S (*),S (*),R (*))-, (S (*),R (*),S (*))-, and (R (*),S (*),S (*))-serricornins were calculated from the data above. The observed(3) J HHs of the naturally occurring serricornin, both acyclic and cyclic forms, are fitted best to calcd.(3) J HHs of (4S (*), 6S (*), 7S (*))-acyclic and (3S (*), 5S (*), 6S (*))-cyclic serricornin, respectively, among those of four enantiomeric pairs of serricornin.

The synthesis of a simple analog of periplanone-B with high biological activity is described mainly on the examination of the conformational property of the key intermediate 10-t-butoxy-6-methyldeca-2,6-dienone by molecular mechanics calculation and dynamic NMR spectroscopy

Micro distribution pattern of tetrodotoxin (TTX) in several tissues of marine puffers Takifugu vermicularis and Chelonodon patoca was investigated by means of monoclonal antibody-based immunoenzymatic technique under light microscope. In the investigation TTX was visualized at glands in the skin of T. vermicularis, while in C. patoca TTX was detected in succiform cells of the skin section. Similarly, in the ovary section of T. vermicularis TTX was recognized at late peri nucleolus stage, yolk granule stage-I, and yolk granule stage-II of oocytes. The oocytes of late peri nucleolus stage and yolk granule stage-I showed TTX antigen at their nucleus and yolk vesicles, while in yolk granule stage-II TTX was visualized at yolk granules and yolk vesicles. In the ovary of C. patoca TTX was detected in the connective tissues and in the nucleus of some perinucleolar oocytes. In the liver and muscle of C. patoca TTX was found to be distributed in parenchymal hepatocytes and muscle fiber, respectively. This study, however, reveals that intra-tissue distribution of TTX varies in respect of species.

A droplet of an oil-in-water emulsion of methyl linoleate in a saccharide or protein solution that contained with a surfactant, a stabilizer, or both was dehydrated by drying equipment for a single droplet that resembled a spray drier. The lipid exposed on the surface of dehydated samples was extracted and measured by gas chromatography. Gum arabic or gelatin without additives resulted in little lipid being exposed; they were good entrapping agents. Little lipid was exposed with a pullulan solution containing lecithin, sugar ester, carboxymethylcellulose, or sodium caseinate but much was exposed with a maltodextrin solution containing any of the surfactants tested. When both the surfactant lecithin and the stabilizer xanthan gum were added to the emulsion prepared in a maltodextrin solution, lipid was not detected. The results suggested that effective entrapping agents of liquid lipids cause much emulsification, stabilize the emulsion (that is, they cause the continuous phase to be very viscous), and create a dehydrated matrix of fine, dense network layers.

The structure of periplanone-A was established by X-ray crystallographic analysis. In comparison of the X-ray and 1H NMR data with those of periplanone-B, it was shown that periphanone-A and -B adopt essentially the same conformation. The epoxy epimer of periplanone-A has l04-times lower biological activity than periplanone-A, and NMR analysis indicated that the molecule exists in a mixture of different conformers. The conformation of the regioisomer, in which the carbonyl and epoxy groups are transposed, was analysed by X-ray crystallography and 1H NMR spectroscopy. It is highly biologically active, and the conformation of the germacranoid skeleton was shown to be almost identical with that of periplanone-A. Simplified analogues and germacrene-D had relatively lower activity. NOE Experiments on these compounds suggested the conformational resemblance of their germacranoid skeletons with those of periplanone-A and -B.

The structure of periplanone-A was established by X-ray crystallographic analysis. In comparison of the X-ray and 1H NMR data with those of periplanone-B, it was shown that periphanone-A and -B adopt essentially the same conformation. The epoxy epimer of periplanone-A has l04-times lower biological activity than periplanone-A, and NMR analysis indicated that the molecule exists in a mixture of different conformers. The conformation of the regioisomer, in which the carbonyl and epoxy groups are transposed, was analysed by X-ray crystallography and 1H NMR spectroscopy. It is highly biologically active, and the conformation of the germacranoid skeleton was shown to be almost identical with that of periplanone-A. Simplified analogues and germacrene-D had relatively lower activity. NOE Experiments on these compounds suggested the conformational resemblance of their germacranoid skeletons with those of periplanone-A and -B.

Bioassays were performed on malePeriplaneta americana L. using synthetic (−)-periplanone-B (P-B) and Hauptmann's (−)-periplanone-A (P-A), and their mixtures at various ratios to estimate the roles of both periplanones for the sexual communication of the species. Both P-A and P-B elicited qualitatively the same responses, such as antennal movement, rapid locomotion, wing raising, and homosexual behavior of male cockroaches, but the threshold of the pheromone activities for P-B was two orders of magnitude lower than that of P-A. Neither synergistic nor inhibitory but only a simple integrated effect on the responses was observed when mixtures of P-A and P-B were applied.