Analysis of the MYD88 L265P mutation in IgM monoclonal gammopathy by semi-nested polymerase chain reaction-based restriction fragment length polymorphism method

MYD88 L265P mutation causes constitutive activation of NF-κB and possible driver mutation in B-cell lymphoid malignancies. It is frequently detected in Waldenstrom’s macroglobulinemia （WM） （50％-100％）, and its detection is important in diagnostic and therapeutic targets of this syndrome. Standard detection method of MYD88 L265P mutation in clinical practice has yet to be established. We developed semi-nested PCR-based restriction fragment length polymorphism （snPCR-RFLP） to detect the mutation. The snPCR-RFLP method is a modification of the PCR-RFLP method, which uses the restriction enzyme BsiEI that recognizes CGACT/CG, intending to increase detection sensitivity by amplification of mutated allele in the DNA sample using semi-nested PCR before enzyme digestion. The detection sensitivity of snPCR-RFLP was estimated as 0.1％, by detecting mutated allele in wild-type allele in the cloned plasmid DNA, which is comparable with allele-specific （AS） PCR method widely used as sensitive detection method. By analyzing 40 cases with IgM monoclonal gammopathy, snPCR-RFLP detected 29/40 （70％） of all cases, 22/31 （70.9％） of WM, and 6/9 （66.6%） of IgM-type monoclonal gammopathy with undetermined significance （IgMMGUS）, including five cases （three cases of WM and two cases of IgMMGUS） in which the mutation was detected only by snPCR-RFLP but not by Sanger sequencing method. Regarding DNA sample status, particularly five cases, a case was extracted from formalin-fixed paraffin-embedded tissue and four cases were extracted from cells by Ficoll-Hypaque density gradient. In correlation with clinical features, the MYD88 mutation detected by snPCR-RFLP method was associated with the adverse prognostic index （WMIPSS） of WM using patient age, hemoglobin （Hb） level, platelet count, β2MG level, and serum IgM level （p＝0.055）. The snPCR-RFLP method is a clinically useful MYD88 mutation detection method that can be performed in general laboratories

A Cluster of Respiratory Syncytial Virus Infections in a Hospital Ward for Adult Immunocompromised Patients

Four male patients admitted to the same ward in the first half of September 201Y were identified to have respiratory syncytial virus（RSV）infection. Their ages ranged from 49 to 85 years（median 72.5）. One patient was infected with human immunodeficiency virus and three patients had hematological malignancies. Following immuno-chromatological testing with a nasal cavity swab, RSV infection was diagnosed. Although blood and sputum cultures were performed in three patients, no significant bacteria were detected. Two cases responded to supportive therapy. However, one patient died secondary to multiple myeloma, and another patient developed pneumonia and died with an exacerbation of leukemia. RSV infections in immunocompromised hosts are associated with a poor prognosis. Early diagnosis will facilitate isolation of infected individuals to prevent hospital outbreaks

生体腎移植の当事者に生じる心理社会的・倫理的課題と、診療に対する提言

学位の種別: 課程博士審査委員会委員 : （主査）東京大学教授 小山 博史, 帝京大学医学部附属病院泌尿器科講師 川合 剛人, 東京大学准教授 神出 誠一郎, 東京大学連携准教授 高山 智子, 東京大学講師 高木 大

Effect of Brønsted Acidity on Ion Conduction in Fluorinated Acetic Acid and N-Methylimidazole Equimolar Mixtures as Pseudo-protic Ionic Liquids

To clarify proton conduction mechanism in protic ionic liquids (PILs) and pseudo-PILs (pPILs), equimolar mixtures of N-methylimidazole (C(1)Im) with fluorinated acetic acids were investigated by Raman spectroscopy, X-ray scattering, and dielectric relaxation spectroscopy (DRS). Only the ionic species exist in the equimolar mixture of C(1)Im and HTFA (HTFA: trifluoroacetic acid). On the other hand, the equimolar mixture of C(1)Im and HDFA (HDFA: difluoroacetic acid) consists of both ionic and electrically neutral species. In particular, not only the electrostatic but also van der Waals interactions with the F atoms were observed in the liquid structures of both [C(1)hIm(+)][TFA(-)] and [C(1)hIm(+)][DFA(-)]. The concept for proton conduction mechanism that we have proposed in previous study was revisited; the proton conduction mechanism could be classified with two linear free energy relationship lines for proton exchange reaction and translation/rotation of proton carriers. Our results exhibit that the proton conduction mechanism changes from proton hopping to vehicle mechanism with increasing acidity of an acid HA in PILs

Possible Proton Conduction Mechanism in Pseudo-Protic Ionic Liquids: A Concept of Specific Proton Conduction

In a previous work, we have found that the pseudo-protic ionic liquid N-methylimidazolium acetate, [C(1)HIm] [OAc] or [Hmim] [OAc], mainly consists of the electrically neutral molecular species N-methylimidazole, C(1)Im, and acetic acid, AcOH, even though the mixture has significant ionic conductivity. This system was revisited by employing isotopic substitution Raman spectroscopy (ISRS) and pulsed field gradient (PFG) NMR self-diffusion measurements. The ISRS and PFG-NMR results obtained fully confirm our earlier findings. In particular, the self-diffusion coefficient of the hydroxyl hydrogen atom in AcOH is identical to that of the methyl hydrogen atoms within the experimental uncertainty, consistent with very little ionization. Therefore, a proton conduction mechanism similar to the Grotthuss mechanism for aqueous acid solutions is postulated to be responsible for the observed electrical conductivity. Laity resistance coefficients (r(ij)) are calculated from the transport properties, and the negative values obtained for the like-ion interactions are consistent with the pseudo-ionic liquid description, that is, the mixture is indeed a very weak electrolyte. The structure and rotational dynamics of the mixture were also investigated using high-energy X-ray total scattering experiments, molecular dynamics simulations, and dielectric relaxation spectroscopy. Based on a comparison of activation energies and the well-known linear free energy relationship between the kinetics and thermodynamics of autoprotolysis, we propose for [C(1)HIm] [OAc] a Grotthus-type proton conduction mechanism involving fast AcOH/AcO- rotation as a decisive step

Comparative analysis of plant isochorismate synthases reveals structural mechanisms underlying their distinct biochemical properties

Isochorismate synthase (ICS) converts chorismate into isochorismate, a precursor of primary and secondary metabolites including salicylic acid (SA). SA plays important roles in responses to stress conditions in plants. Many studies have suggested that the function of plant ICSs is regulated at the transcriptional level. In Arabidopsis thaliana, the expression of AtICS1 is induced by stress conditions in parallel with SA synthesis, and AtICS1 is required for SA synthesis. In contrast, the expression of NtICS is not induced when SA synthesis is activated in tobacco, and it is unlikely to be involved in SA synthesis. Studies on the biochemical properties of plant ICSs are limited, compared with those on transcriptional regulation. We analyzed the biochemical properties of four plant ICSs: AtICS1, NtICS, NbICS from Nicotiana benthamiana, and OsICS from rice. Multiple sequence alignment analysis revealed that their primary structures were well conserved, and predicted key residues for ICS activity were almost completely conserved. However, AtICS1 showed much higher activity than the other ICSs when expressed in Escherichia coli and N. benthamiana leaves. Moreover, the levels of AtICS1 protein expression in N. benthamiana leaves were higher than the other ICSs. Construction and analysis of chimeras between AtICS1 and OsICS revealed that the putative chloroplast transit peptides (TPs) significantly affected the levels of protein accumulation in N. benthamiana leaves. Chimeric and point-mutation analyses revealed that Thr(531), Ser(537), and Ile(550) of AtICS1 are essential for its high activity. These distinct biochemical properties of plant ICSs may suggest different roles in their respective plant species.ArticleBioscience Reports 38(2) : BSR20171457, (2018)journal articl

Dynamic Chelate Effect on the Li+-Ion Conduction in Solvate Ionic Liquids

Lithium-glyme solvate ionic liquids (Li-G SILs), which typically consist of a lithium-ion (Li+) solvated by glymes of oligoethers and its counter anion, are expected as promising electrolytes for lithium secondary batteries. Additionally, a specific ligand-exchange Li+ conduction mechanism was proposed at the electrode/electrolyte interface of the cell using Li-G SILs. To reveal Li+ conduction in SILs, Li-G SILs with varying ethylene oxide chain lengths were investigated using various techniques that are sensitive to solution structure and dynamics. We found good correlations between the relaxation time of the slowest dielectric mode and the ionic conductivity as well as viscosity. We propose that a dynamic chelate effect, which is closely related to solvent exchange and/or contact ion-pair formation/dissociation, is important for Li+ conduction in these Li-G SILs

Geological history of the land area between Okinawa Jima and Miyako Jima of the Ryukyu Islands, Japan, and its phylogeographical significance for the terrestrial organisms of these and adjacent islands

Abstract The modern and Late Pleistocene terrestrial fauna of Miyako Jima and adjacent islands (the Miyako Islands) in the southern Ryukyu Islands, southwestern Japan, includes some endemic taxa or genetically unique populations that exclusively have closest allies in the more isolated Okinawa Jima and adjacent islands (the Okinawa Islands) than in the Yaeyama Islands, which are located southwest of the Miyako Islands with much narrower intervening straits. Those taxa or populations include representatives of lineages that have physiologically highly limited ability for over-sea dispersal and the Miyako Islands are currently separated from the Okinawa Islands by at least 300 km of open water; therefore, the formation of this phylogeographical pattern is perplexing. In this study, we review the late Cenozoic geology of the Miyako Islands, southern Okinawa Jima, the Okinawa–Miyako submarine plateau (OMSP; a plateau located between Okinawa Jima and Miyako Jima), and the Kerama gap, which is a depression between the OMSP and Okinawa Jima. We then consider the origin of the modern and Late Pleistocene terrestrial animals, including a number of non-volant vertebrates on the Miyako Islands. Finally, we propose a new hypothesis (the OMSP hypothesis) to explain the enigmatic composition of modern and Late Pleistocene terrestrial vertebrate fauna of the islands. Southern Okinawa Jima was uplifted and emerged after ca. 2 Ma and was temporarily connected to the OMSP, which is likely to have emerged earlier than southern Okinawa Jima, to form a large island extending from Okinawa Jima to the Miyako Islands with a NE–SW direction of ~ 400 km. Subsequently, Okinawa Jima became separated from the OMSP when the Ryukyu Group—which is composed of Quaternary reef and associated fore-reef and shelf deposits—began to accumulate around the island at 1.7–1.4 Ma. During the interval from 2.0 to 1.7–1.4 Ma, numerous terrestrial animals, including flightless vertebrates, extended their distribution to the OMSP. Although the Miyako Islands repeatedly underwent complete submergence during deposition of the main part of the Ryukyu Group (1.25–0.4 Ma), they were uplifted and emerged to become a land area after ca. 0.4 Ma. In contrast, the OMSP subsided after ca. 0.4 Ma and was almost completely submerged after 0.27 Ma. During ca. 0.4–0.27 Ma, terrestrial animals migrated from the OMSP to the Miyako Islands

Change in N-Glycosylation of Plasma Proteins in Japanese Semisupercentenarians.

An N-glycomic analysis of plasma proteins was performed in Japanese semisupercentenarians (SSCs) (mean 106.7 years), aged controls (mean 71.6 years), and young controls (mean 30.2 years) by liquid chromatography/mass spectrometry (LC/MS) using a graphitized carbon column. Characteristic N-glycans in SSCs were discriminated using a multivariate analysis; orthogonal projections to latent structures (O-PLS). The results obtained showed that multi-branched and highly sialylated N-glycans as well as agalacto- and/or bisecting N-glycans were increased in SSCs, while biantennary N-glycans were decreased. Since multi-branched and highly sialylated N-glycans have been implicated in anti-inflammatory activities, these changes may play a role in the enhanced chronic inflammation observed in SSCs. The levels of inflammatory proteins, such as CRP, adiponectin, IL-6, and TNF-α, were elevated in SSCs. These results suggested that responses to inflammation may play an important role in extreme longevity and healthy aging in humans. This is the first study to show that the N-glycans of plasma proteins were associated with extreme longevity and healthy aging in humans

Stratified layer analysis reveals intrinsic leptin stimulates cryptal mesenchymal cells for controlling mucosal inflammation

Mesenchymal cells in the crypt play indispensable roles in the maintenance of intestinal epithelial homeostasis through their contribution to the preservation of stem cells. However, the acquisition properties of the production of stem cell niche factors by the mesenchymal cells have not been well elucidated, due to technical limitations regarding the isolation and subsequent molecular and cellular analyses of cryptal mesenchymal cells. To evaluate the function of mesenchymal cells located at the large intestinal crypt, we established a novel method through which cells are harvested according to the histologic layers of mouse colon, and we compared cellular properties between microenvironmental niches, the luminal mucosa and crypts. The gene expression pattern in the cryptal mesenchymal cells showed that receptors of the hormone/cytokine leptin were highly expressed, and we found a decrease in Wnt2b expression under conditions of leptin receptor deficiency, which also induced a delay in cryptal epithelial proliferation. Our novel stratified layer isolation strategies thus revealed new microenvironmental characteristics of colonic mesenchymal cells, including the intrinsic involvement of leptin in the control of mucosal homeostasis