Luteibacter jiangsuensis blood stream infection: a first case report.

BACKGROUND: Luteibacter jiangsuensis is a gram-negative aerobic bacillus that was first isolated from soil samples at a pesticide factory in China and reported in 2011. Here, we describe the first case of L. jiangsuensis infection in human. CASE PRESENTATION: A 59-year-old Japanese woman undergoing treatment for Crohn's disease was admitted to our hospital with fever. Clinical examination indicated catheter-related bloodstream infection. The catheter was removed and meropenem was initiated. Morphologically identical glucose non-fermentative gram-negative bacilli were detected from two sets of aerobic blood culture and catheter-tip cultures. MALDI-TOF mass spectrometry failed to identify the bacterium, which was later identified as L. jiangsuensis by 16 S rRNA gene sequencing. Antimicrobial susceptibility test revealed that the isolate was resistant to carbapenem, therefore meropenem was switched to intravenous levofloxacin (500 mg/day). After 14 days of treatment with levofloxacin, the patient was discharged. CONCLUSIONS: This is the first case of L. jiangsuensis infection in human. The strain was identified by 16 S rRNA gene sequence analysis.

Dysregulation of humoral immunity in Foxp3 conditional-knockout mice.

Foxp3+ regulatory T cells (Tregs) are crucial for maintaining tolerance to self-antigens and preventing autoimmune diseases. Loss of Foxp3 expression leads to autoimmunity and disrupts humoral immune responses, including hyperproduction of immunoglobulin E (IgE). Elucidation of how Tregs control antibody production can lead to the development of new therapies for autoimmune and allergic diseases. However, premature death of Foxp3-deficient mice makes it difficult to analyze the roles of Tregs in humoral immunity of adult mice. In this study, we developed Foxp3 conditional-knockout mice (Foxp3floxR26CreERT2) in which the Foxp3 gene was inducibly deleted by tamoxifen administration. After oral administration of tamoxifen, titers of immunoglobulins, particularly IgG2c and IgE, were increased in Foxp3floxR26CreERT2 mice compared with that in controls. Under these conditions, CD4+ T cells from Foxp3floxR26CreERT2 mice had increased expression of several activation markers, including inducible costimulator and CD40 ligand, as well as the cytokines interleukin 4 and interferon gamma. In addition, the proportions of T follicular helper (Tfh) cells and germinal center (GC) B cells were increased in Foxp3floxR26CreERT2 mice compared with those in controls. These results indicated that Tregs controlled excessive or pathogenic antibody production by suppressing Tfh cell differentiation and GC formation. Furthermore, these data suggested that Foxp3floxR26CreERT2 mice could be a useful tool for screening therapeutic agents.

Water Filling and Emptying Kinetics in Two-Dimensional Hexagonal Mesoporous Silica of the Same Pore Diameter but Different Pore Lengths.

The effect of pore length on the water filling and emptying rates was studied using mesoporous silica (MPS) with same pore diameter but different pore lengths. The pore diameter of the synthesized MPS was ∼8 nm, whereas the average pore lengths were 460, 1,770, and 4000 nm. The gravimetric method was employed to record the time course of the adsorbed mass of water in MPS at 298 K and 1 atm. In both the filling and emptying processes, the relaxation curves (time course of adsorbed mass of water per unit mass of sample) were not significantly related to the pore length. This independence of the initial adsorption and desorption rates on the pore length suggests that the surface of the MPS aggregates is the bottleneck in the overall adsorption and desorption processes and that the initial mass flux in each nanopore is inversely proportional to the pore length. Furthermore, because the relaxation times to reach the equilibrium state were independent of the pore length, the mass flux of water uptake, release, and transport probably increase with an increase in the pore length during the entire adsorption and desorption processes. A transport model to describe these phenomena was proposed.

A novel mouse model for tracking the fate of CXCR5-expressing T cells.

The germinal center (GC) reaction, a critical process in the humoral immune response, requires follicular helper T (Tfh) cells. Tfh cells express the master transcription factor Bcl6 and chemokine receptor CXCR5, which enable them to migrate from the T cell zone to B cell follicles and interact with GC B cells. However, CXCR5 is downregulated when Tfh cells become memory cells. Therefore, it is difficult to track Tfh cells continuously in vivo. In this study, we generated a mouse strain, Cxcr5CreERT2R26Tomato, in which the fluorescent protein tdTomato is inducibly expressed in CXCR5+ cells by tamoxifen administration. After the oral administration of tamoxifen, most Tfh cells in Peyer's patches (PP) from Cxcr5CreERT2R26Tomato mice were tdTomato+. To track antigen-specific Tfh cells in vivo, OVA-specific OT-II T cells derived from Cxcr5CreERT2R26Tomato mice were transferred to wild-type mice, and the recipient mice were immunized with OVA followed by tamoxifen administration. CXCR5+ T cells became tdTomato+ and were mainly located in B cell follicles and GC areas 8 days after immunization. Four weeks after immunization, tdTomato+ OT-II T cells migrated from B cell follicles to the T-B border area and T cell zone after CXCR5 downregulation and CCR7 upregulation. These results indicate that Cxcr5CreERT2R26Tomato mice are a useful tool for studying the cell fate of differentiated Tfh cells in vivo and therefore have implications for the development of therapeutic strategies for infectious and autoimmune diseases.

Vapor-Phase Synthesis of ZIF-8 MOF Thick Film by Conversion of ZnO Nanorod Array.

ZIF-8 metal organic framework "micrometer-thick" films were constructed from ZnO precursor by a vapor-phase synthesis. The ZnO-to-ZIF-8 crystal transformation proceeded in the presence of 2-methylimidazole (Hmim) vapor. Continuous coatings of intergrown ZIF-8 crystals require control of a nucleation density. The dependence of ZnO crystal plane on the ZnO-to-ZIF-8 crystal transformation was investigated using four bulk ZnO single crystals: a-plane (11-20), c-plane (0001), m-plane (10-10), and r-plane (10-11). It was revealed that the m-plane (10-10) of ZnO is more effectively transformed into ZIF-8. In this work, highly oriented ZnO nanorod array film was used to provide the transport pathway of Hmim molecules and volume expansion space of ZnO-to-ZIF-8 crystal transformation for nucleation and crystal intergrowth. The high conversion of ZnO nanorod array into ZIF-8 in a short time could be achieved because (1) such mass transfer is easy due to the uniform internanorod distance being maintained during reaction and (2) the surface of the nanorod array is dominated by the highly reactive m-plane (10-10).

Exome QTL-seq maps monogenic locus and QTLs in barley.

BACKGROUND: QTL-seq, in combination with bulked segregant analysis and next-generation sequencing (NGS), is used to identify loci in small plant genomes, but is technically challenging to perform in species with large genomes, such as barley. A combination of exome sequencing and QTL-seq (exome QTL-seq) was used to map the mono-factorial Mendelian locus black lemma and pericarp (Blp) and QTLs for resistance to net blotch disease, a common disease of barley caused by the fungus Pyrenophora teres, which segregated in a population of 100 doubled haploid barley lines. METHODS: The provisional exome sequences were prepared by ordering the loci of expressed genes based on the genome information and concatenating genes with intervals of 200-bp spacer "N" for each chromosome. The QTL-seq pipeline was used to analyze short reads from the exome-captured library. RESULTS: In this study, short NGS reads of bulked total DNA samples from segregants with extreme trait values were subjected to exome capture, and the resulting exome sequences were aligned to the reference genome. SNP allele frequencies were compared to identify the locations of genes/QTLs responsible for the trait value differences between lines. For both objective traits examined, exome QTL-seq identified the monogenic Mendelian locus and associated QTLs. These findings were validated using conventional mapping approaches. CONCLUSIONS: Exome QTL-seq broadens the utility of NGS-based gene/QTL mapping in organisms with large genomes.

Characterization of Rab-interacting lysosomal protein in the brain of Bombyx mori.

Rab guanosine triphosphatases in eukaryotic cells are key regulators of membrane-trafficking events, such as exocytosis and endocytosis. Rab7 regulates traffic from early to late endosomes and from late endosomes to vacuoles/lysosomes. The Rab7-interacting lysosomal protein (RILP) was extracted from the silkworm, Bombyx mori (B. mori), and expressed in Escherichia coli (E. coli), followed by its purification. The glutathione sulfotransferase pull-down assay revealed that Rab7 of B. mori interacted with RILP of B. mori. We then produced antibodies against RILP of B. mori in rabbits for their use in Western immunoblotting and immunohistochemistry. Western immunoblotting of brain tissue for RILP revealed a single band, at approximately 50 kD. RILP-like immunohistochemical reactivity (RILP-ir) was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Furthermore, RILP-ir was colocalized with the eclosion hormone-ir and bombyxin-ir. However, RILP-ir was not colocalized with prothoracicotropic hormone-ir. These results were similar to those of Rab7 from our previous study. These findings suggest that RILP and Rab7 are involved in the neurosecretion in a restricted subtype of neurons in B. mori. Thus, our study is the first to report of a possible relationship between an insect Rab effector and neurosecretion.

3'-ethynylcytidine, an RNA polymerase inhibitor, combined with cisplatin exhibits a potent synergistic growth-inhibitory effect via Vaults dysfunction.

BACKGROUND: We previously reported that 3'-ethynylcytidine (ECyd, TAS-106), an RNA polymerases inhibitor, enhances the anti-tumor efficacy of platinum in several tumor types in both in vitro and in vivo tumor models. However, the molecular mechanisms underlying the ECyd-induced enhancement remain elusive. METHODS: Cisplatin (CDDP)-resistant head and neck cancer KB cells were established by stepwise dose escalation with CDDP. The combination effect of ECyd and CDDP were assessed using isobologram analysis. The transcriptional and post-translational statuses of several molecules were detected using real-time PCR, immunoblot analysis and immunocytochemistry. Xenograft assays were used to confirm the mechanisms underlying the ECyd induced enhancement of CDDP anti-tumor efficacy in vivo. RESULTS: ECyd sensitized KB to CDDP by inhibiting the drug transporter Vault complex (Vaults). First, we showed that Vaults were overexpressed in CDDP-resistant KB cells. The suppression of major vault protein (MVP) by RNA interference restored the sensitivity to CDDP. Next, we showed that ECyd significantly sensitized the resistant cells to CDDP, compared with the parental paired cell line. A molecular analysis revealed that ECyd inhibited the synthesis of vRNAs as well as the induction of MVP, both of which are critical components of Vaults as a drug transporter. Furthermore, we found that the synergistic effect of ECyd and CDDP was correlated with the MVP expression level when the effect was analyzed in additional cancer cell lines. Finally, we demonstrated that ECyd decreased the vRNAs expression level in xenograft tumor. CONCLUSIONS: Our data indicated the ability of ECyd to cancel the resistance of cancer cells to CDDP by inhibiting the Vaults function and the decrease of Vaults expression itself, and the ability of the combination therapy with CDDP and ECyd to offer a new strategy for overcoming platinum resistance. Moreover, the study results suggest that Vaults could be a biomarker for stratifying patients who may benefit from the combination therapy with ECyd and platinum.

Study of optical rotation based on the molecular structure in fused oligomers of macrocycles.

We designed a unique oligomer form in which several helically twisted macrocycles (M- or P-helicity) are arranged through fusion. We investigated the optical rotation of a series of fused oligomers of macrocycles with a difference in the number and arrangement of elements associated with point-chiral auxiliary. Some oligomers cooperatively attained a situation where an identical sense of twisting was preferred throughout the entire molecule. On the basis of these results, we estimated diastereomeric excess induced in each oligomer. We revealed that the molar optical rotation per element was modulated with a rotational angle between elements: an increase via 0° rotational arrangement, a decrease via 180° rotational arrangement, or a decrease via cyclic arrangement. Alternatively, for other oligomers in which several diastereomeric conformers coexist, we uniquely attempted to consider the optical rotation based on the molecular structure through the assessment of a change ratio of the absorption dissymmetry factor before and after complexation with an achiral guest. We found that molar optical rotation could be different based on the arrangement, although actual measured values were similar.

Relationship between the expression of Rab family GTPases and neuropeptide hormones in the brain of Bombyx mori.

Rab proteins are small GTPases that play essential roles in vesicle transport. In this study, we examined the expression of Rab proteins and neuropeptide hormones in the brain of the silkworm, Bombyx mori. We produced antibodies against B. mori Rab1 and Rab14 in rabbits. Immunoblotting of samples of brain tissue from B. mori revealed a single band for each antibody. Rab1 and Rab14 immunohistochemical labeling in the brain of B. mori was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Rab1, Rab7 and Rab14 co-localized with bombyxin. Rab1 and Rab7 co-localized with eclosion hormone. Rab1 co-localized with prothoracicotropic hormone. These results suggest that Rab1, Rab7 and Rab14 may be involved in neuropeptide transport in the brain of B. mori. This is the first report on the specificity of Rab proteins for the secretion of different neuropeptides in insects.

Spatialising urban health vulnerability: An analysisof NYC's critical infrastructure during COVID-19.

This paper examines how fragmentation of critical infrastructure impacts the spread of the coronavirus outbreak in New York City at the neighbourhood level. The location of transportation hubs, grocery stores, pharmacies, hospitals and parks plays an important role in shaping spatial disparities in virus spread. Using supervised machine learning and spatial regression modelling we examine how the geography of COVID-19 case rates is influenced by the spatial arrangement of four critical sectors of the built environment during the public health emergency in New York City: health care facilities, mobility networks, food and nutrition and open space. Our models suggest that an analysis of urban health vulnerability is incomplete without the inclusion of critical infrastructure metrics in dense urban geographies. Our findings show that COVID-19 risk at the zip code level is influenced by (1) socio-demographic vulnerability, (2) epidemiological risk, and (3) availability and access to critical infrastructure.

Effect of abutment tooth location on the accuracy of digital impressions obtained using an intraoral scanner for removable partial dentures.

PURPOSE: To verify the effect of abutment tooth location on the accuracy of digital impressions obtained using an intraoral scanner (IOS) for removable partial dentures (RPDs). METHODS: The target abutment teeth included the left first premolar (#34), second molar (#37), and right second premolar (#45) in a mandibular Kennedy class II model and the left and right second molars (#37, #47) in a class III model. Only #37 was isolated from the remaining teeth by the mucosal area in both models. Digital impressions were obtained using a desktop scanner (reference data) and an IOS (IOS data; scanning origin #37; n=10). The general trueness based on the entire model superimposition (TG), local trueness (TL) of an individual tooth, and dimensional accuracy (coordinate and linear accuracy) of the IOS data of the target abutment teeth were compared (α=0.05). RESULTS: In both models, #37 showed significantly inferior TG (P<0.01), superior TL (P<0.01), and mesial coordinate displacement (P<0.01 and P<0.05 in class II and III models, respectively). Intra-model comparisons showed that #45 in the class II model and #47 in the class III model had significantly inferior linear accuracy (P<0.05 and P<0.01, respectively) and buccal coordinate displacement (P<0.05 and P<0.01, respectively) compared with the other target teeth. CONCLUSIONS: In digital impressions of RPDs, isolation of abutment teeth by mucosal areas can reduce general trueness based on the entire dental arch and mesial tooth displacement, whereas increased distance from the scanning origin can adversely affect local trueness and dimensional accuracy.

Morphological Comparison of Residual Ridge in Impression for Removable Partial Denture between Digital and Conventional Techniques: A Preliminary In-Vivo Study.

Although digital impression using an intraoral scanner (IOS) has been applied for removable partial denture (RPD) fabrication, it is still unclear how the morphology of a residual ridge recorded by digital impression would differ from that recorded by conventional impression. This in vivo study investigated the morphological difference in the recorded residual ridge between digital and conventional impressions. Vertical and horizontal displacements (VD and HD) in residual ridges recorded by digital and conventional impressions were assessed in 22 participants (15 female; mean age 78.2 years) based on the morphology of the tissue surface of in-use RPD. Additionally, the mucosal thickness of the residual ridge was recorded using an ultrasound diagnostic device. VD and HD were compared using the Wilcoxon signed-rank test, and the correlation of mucosal thickness with VD and HD was analyzed using Spearman's ρ. The VD of digital impression was significantly greater than that of a conventional impression (p = 0.031), while no significant difference was found in HD (p = 0.322). Meanwhile, the mucosal thickness showed no significant correlation with the recorded morphology of the residual ridge, regardless of the impression techniques. It was concluded that the digital impression would result in a greater displacement in the height of the residual ridge from the morphology of in-use RPD than the conventional impression.

Effect of Scanning Origin Location on Data Accuracy of Abutment Teeth Region in Digital Impression Acquired Using Intraoral Scanner for Removable Partial Denture: A Preliminary In Vitro Study.

The aim of this study was to investigate the effect of scanning origin location on the data accuracy of removable partial denture (RPD) abutment teeth region in digital impressions acquired by an intraoral scanner. A mandibular partially edentulous model including the following target abutment teeth was used: the left second molar (#37); left first premolar (#34); and right second premolar (#45). The following scanning strategies were tested: the strategy starting from #37 to mesial direction (37M); strategies starting from #34 to mesial (34M) and distal directions (34D), and strategies starting from #45 to mesial (45M) and distal directions (45D). The evaluated measures were trueness, precision, and linear accuracy. One-way and two-way ANOVA were performed for the comparison of trueness and linear accuracy, while Kruskal-Wallis test was performed for the precision comparison (α = 0.05). 45M and 45D showed significantly superior trueness of #34 to 37M and 34D. 45M also showed significantly superior trueness of #45 to 34. 45D showed significantly inferior linear accuracy of #34 and superior linear accuracy of #45 compared to other strategies. It was concluded that scanning origin location would have an impact on data accuracy of RPD abutment teeth region in digital impressions acquired by intraoral scanner.

Multivalent cation and polycation polymer preparations influence pharmacokinetics of dolutegravir via chelation-type drug interactions.

Dolutegravir (DTG) is an integrase inhibitor, whose gastrointestinal absorption is impaired by the formation of chelates with multivalent metal cation preparations. However, little is known regarding the interactions of DTG with preparations containing other multivalent metal cations or with polycation polymer preparations. This study examined how the pharmacokinetics of DTG are affected by co-administration with Al(OH)3, LaCO3, and the polycation polymers bixalomer (Bxl) and sevelamer (Svl). Prior to oral administration of DTG (5 mg/kg), rats were orally administered Al(OH)3 (150 or 300 mg/kg), LaCO3 (50 or 75 mg/kg), Bxl (250 or 500 mg/kg), or Svl (300 or 600 mg/kg). Serum concentrations of DTG were then measured over the next 24 h. Compared to the administration of DTG alone, its co-administration with Al(OH)3, LaCO3, Bxl, and Svl led to reduced serum concentration of DTG, and consequently, a significantly reduced area under the curve. These comparisons also revealed a considerable reduction in the maximum concentration, suggesting that the interactions of these agents with DTG in the intestinal tract inhibit absorption of DTG. The above results demonstrate that Al(OH)3, LaCO3, Bxl, and Svl affect the pharmacokinetics of DTG and indicate the need for caution when combining any of the above preparations with DTG.

Influence of cytidine deaminase on antitumor activity of 2'-deoxycytidine analogs in vitro and in vivo.

Antitumor 2'-deoxycytidine (dCyd) analogs such as gemcitabine (dFdC), cytarabine (Ara-C), and 2'-C-cyano-2'-deoxy-1-ß-d-arabinofuranosylcytosine (CNDAC) are activated by dCyd kinase, whereas cytidine deaminase (CDA) inactivates them by conversion to their uracil forms. To elucidate the relationship between the chemosensitivity to antitumor dCyd nucleosides and CDA expression, we established a stable line of human gastric carcinoma TMK-1 cells constitutively overexpressing CDA (TMK-1/CDA) and examined its chemosensitivity to antitumor dCyd analogs in vitro and in vivo. We observed comparable reactivity for dFdC and Ara-C, and the substrate reactivity of CNDAC to recombinant human CDA was more than 10 times less efficient than those of Ara-C and dFdC. Next, we examined the in vitro chemosensitivity of TMK-1/CDA and observed a marked decrease in the sensitivity of TMK-1/CDA to Ara-C, dFdC, and CNDAC compared with mock-transfected cells. In addition, we transplanted TMK-1/CDA cells into a nude mouse xenograft model and examined their in vivo chemosensitivity to CNDAC. The in vivo antitumor effect of CNDAC on TMK-1/CDA cells was substantially reduced compared with that of mice transplanted with mock-transfected cells. These results indicate that CDA could play an important role in regulating susceptibility to antitumor dCyd analogs in vitro and in vivo. In addition, the expression level of CDA was found to affect the antitumor activity of CNDAC, even though the substrate reactivity of CNDAC to CDA is relatively low.

Concurrent administration with multivalent metal cation preparations or polycationic polymer preparations inhibits the absorption of raltegravir via its chelation.

OBJECTIVES: Raltegravir (RAL) that can form chelates with multivalent metal cations shows lateral interactions with multivalent metal cation and polycationic polymer. We investigated the interactions of RAL with multivalent metal cation preparations, Al(OH)3 and LaCO3 , and polycationic polymer preparations, bixalomer (Bxl) and sevelamer (Svl). METHODS: Immediately before the oral administration of 40 mg/kg RAL, the rats were administered orally with the vehicle, Al(OH)3 , LaCO3 , Bxl, or Svl, and the time course of RAL serum concentration was followed. The in vitro binding affinity of RAL with multivalent metal cation and polycationic polymer was also evaluated using isothermal titration calorimetry (ITC). RESULTS: When Al(OH)3 , LaCO3 , Bxl, or Svl was concomitantly administered with RAL, the maximum concentration and area under the curve were significantly lower than those when RAL was administered alone. ITC showed the interaction of RAL with Al(OH)3 as an enthalpy-driven reaction and its interactions with LaCO3 and Bxl as entropy-enthalpy mixed reactions. CONCLUSIONS: The interaction of RAL with Al(OH)3 , LaCO3, Bxl, or Svl can inhibit RAL absorption into the gastrointestinal tract, and thus, the multivalent metal cation and polycationic polymer are the modifying factors that can affect RAL pharmacokinetics.

Dual dynamic chirality generated in the assembly of three achiral rods through the three-fold twisting of a macrocycle.

We demonstrated dynamic chirality based on assemblies of three achiral rods with a twisted macrocycle. The three-fold twisting of a macrocycle can lead to two different chiral forms with C2- or D3-symmetry. Through a transmission of chirality to each dynamic chiral form, a helical-sense preference was successfully induced.

Chiral diversification through the assembly of achiral phenylacetylene macrocycles with a two-fold bridge.

We demonstrate so-called "chiral diversification", which is a design strategy to create multiple chiral molecules through the assembly and double-bridging of achiral components. We used phenylacetylene macrocycles (PAMs) as an achiral element. In a molecule, two achiral rings of [6]PAM are stacked one above the other, or bound to each other mechanically. As an alternative, a single enlarged ring of [12]PAM was also assumed to be a doubled form of [6]PAM. In any case, one or two ring(s) are doubly-bridged by covalent bonds to exert chirality. Through intramolecular two-bond formation, these multiple chiral molecules were obtained as a set of products in one reaction. The dynamic chirality generated in molecules with either two helically-stacked rings of [6]PAM or a single helically-folded ring of [12]PAM was characterized by induced Cotton effects with the aid of an external chiral source. Thus, a chiral structure based on [12]PAM could be demonstrated as the first success. Alternatively, enantiomeric separation was achieved for molecules with two interlocked rings of [6]PAM to show remarkable chiroptical properties.

Folylpolyglutamate synthase and gamma-glutamyl hydrolase regulate leucovorin-enhanced 5-fluorouracil anticancer activity.

Although 5-fluorouracil (5-FU) plus leucovorin (LV) is a standard chemotherapy regimen for colorectal cancer, the factors that determine the LV-mediated enhancement of the antitumor activity of 5-FU have remained unknown. We investigated the roles of folylpolyglutamate synthase (FPGS) and gamma-glutamyl hydrolase (GGH), which are the main enzymes involved in folate metabolism, in the effect of LV. LV enhanced the anticancer activity of 5-FU and the level of reduced folate in human colon cancer cells. Small-interfering RNA (siRNA) transfected into DLD-1 cells to downregulate FPGS reduced the basal level of reduced folate, the folate level after LV treatment, and the enhancement of 5-fluoro-2'-deoxyuridine (FdUrd)-induced cytotoxicity elicited by LV. By contrast, the downregulation of GGH by siRNA increased cellular sensitivity to FdUrd combined with LV. These results suggest that FPGS and GGH expression levels in tumors are determinants of the efficacy of LV in enhancing the antitumor activity of 5-FU.