<i>Salmonella</i> Flagellum

Flagella-driven motility contributes to effective bacterial invasion. The bacterial flagellum of Salmonella enterica is a rotary motor powered by an electrochemical potential difference of protons across the cytoplasmic membrane. The flagellum is composed of several basal body rings and an axial structure consisting of the rod as a drive shaft, the hook as a universal joint and the filament as a helical propeller. The assembly of the axial structure begins with the rod, followed by the hook and finally the filament. A type III protein export apparatus is located at the flagellar base and transports flagellar axial proteins from the cytoplasm to the distal end of the growing flagellar structure where their assembly occurs. The protein export apparatus coordinates flagellar gene expression with assembly, allowing the hierarchy of flagellar gene expression to exactly parallel the flagellar assembly process. The basal body can accommodate a dozen stator complexes around a rotor ring complex in a load-dependent manner. Each stator unit conducts protons and pushes the rotor. In this book chapter, we will summarize our current understanding of the structure and function of the Salmonella flagellum

Development of the Counseling Center Assessment of Psychological Symptoms‐Japanese version: Pilot study

There is currently no reliable and valid multidimensional instrument for measuring psychological symptoms among Japanese university students. The purpose of this pilot study was to translate the Counseling Center Assessment of Psychological Symptoms‐62 (CCAPS‐62) into Japanese and evaluate its validity and reliability. Following robust translation procedures, the CCAPS‐Japanese was created. In the validation study, 2,758 undergraduate students from 11 universities (mean age = 19.08 ± 1.85 years) completed the CCAPS‐Japanese. The results of confirmatory factor analysis supported the theoretical eight‐factor structure model of the CCAPS‐Japanese with the exclusion of seven items. The decision to retain/remove items was made by evaluating factor loadings and model fit indices while considering cultural equivalence and structural validity. Using the finalized 55‐item CCAPS‐Japanese, further analyses demonstrated that the eight subscales had acceptable to good internal consistencies (α = .61–.89). Thus, the tool’s validity and reliability were established. The CCAPS‐Japanese may be appropriate for assessing the psychological concerns of Japanese university students.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/153629/1/cpp2412_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/153629/2/cpp2412.pd

The Architecture of the Cytoplasmic Region of Type III Secretion Systems

Type III secretion systems (T3SSs) are essential devices in the virulence of many Gram-negative bacterial pathogens. They mediate injection of protein effectors of virulence from bacteria into eukaryotic host cells to manipulate them during infection. T3SSs involved in virulence (vT3SSs) are evolutionarily related to bacterial flagellar protein export apparatuses (fT3SSs), which are essential for flagellar assembly and cell motility. The structure of the external and transmembrane parts of both fT3SS and vT3SS is increasingly well-defined. However, the arrangement of their cytoplasmic and inner membrane export apparatuses is much less clear. Here we compare the architecture of the cytoplasmic regions of the vT3SSs of Shigella flexneri and the vT3SS and fT3SS of Salmonella enterica serovar Typhimurium at ~5 and ~4 nm resolution using electron cryotomography and subtomogram averaging. We show that the cytoplasmic regions of vT3SSs display conserved six-fold symmetric features including pods, linkers and an ATPase complex, while fT3SSs probably only display six-fold symmetry in their ATPase region. We also identify other morphological differences between vT3SSs and fT3SSs, such as relative disposition of their inner membrane-attached export platform, C-ring/pods and ATPase complex. Finally, using classification, we find that both types of apparatuses can loose elements of their cytoplasmic region, which may therefore be dynamic

Cryo-EM structure of a functional monomeric Photosystem I from Thermosynechococcus elongatus reveals red chlorophyll cluster

A high-resolution structure of trimeric cyanobacterial Photosystem I (PSI) from Thermosynechococcus elongatus was reported as the first atomic model of PSI almost 20 years ago. However, the monomeric PSI structure has not yet been reported despite long-standing interest in its structure and extensive spectroscopic characterization of the loss of red chlorophylls upon monomerization. Here, we describe the structure of monomeric PSI from Thermosynechococcus elongatus BP-1. Comparison with the trimer structure gave detailed insights into monomerization-induced changes in both the central trimerization domain and the peripheral regions of the complex. Monomerization-induced loss of red chlorophylls is assigned to a cluster of chlorophylls adjacent to PsaX. Based on our findings, we propose a role of PsaX in the stabilization of red chlorophylls and that lipids of the surrounding membrane present a major source of thermal energy for uphill excitation energy transfer from red chlorophylls to P700

Evidence for suboceanic small-scale convection from a “garnet”-bearing lherzolite xenolith from Aitutaki Island, Cook Islands

海洋マントルにおける小スケール対流の証拠検出 --南太平洋アイツタキ島マントル捕獲岩からのアプローチ --.京都大学プレスリリース. 2024-07-08.Garnet peridotite xenoliths have been rarely reported from suboceanic mantle. Petrographic and geochemical characteristics of garnet-bearing oceanic peridotite xenoliths provide precious information on dynamics of the suboceanic lithosphere and asthenosphere interaction. We examined a lherzolite xenolith included in olivine nephelinite lava from Aitutaki Island, a member of the Cook-Austral volcanic chain. The lherzolite xenolith contains reddish fine-grained (< 5 µm in size) mineral aggregates (FMAs) with size range of 0.5–6 mm, consisting of olivine, calcic and sodic plagioclases, aluminous spinel, native iron, and nepheline. Microstructural observations and chemical data corroborate that the FMA is a decomposed pyrope-rich garnet including chromian spinel grains with an irregular highly indented morphology in the center. The FMA is surrounded by pyroxene-poor and olivine-rich aureole. The spatial and morphological relationships of FMA and chromian spinel with pyroxene-depleted margin suggest a reaction of aluminous spinel + pyroxenes → pyrope-rich garnet + olivine, which requires a compression before decomposition of the garnet to FMA. An orthopyroxene grain shows slight but clear chemical zoning characterized by increase in Al, Ca, and Cr from the grain center to the rim. The zoning patterns of Al and Ca in the orthopyroxene grain can be modeled by diffusion-controlled solid-state reactions induced by pressure and temperature changes, keeping surface concentrations in equilibrium with the other coexisting mineral phases. The results indicate that the mantle, from which the lherzolite xenolith was derived, underwent isothermal decompression followed by a weak heating on a time scale of a few tenths of million years before the xenolith extraction. From the deduced compression and decompression histories, we hypothesize that the mantle beneath Aitutaki Island was once dragged down to a garnet-stable deep mantle region and brought up later by small-scale sublithospheric convection

Promotion of IL-4- and IL-5-dependent differentiation of anti-μ-primed B cells by ascorbic acid 2-glucoside

The stable ascorbic acid derivative 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G) was used to investigate the role of ascorbic acid (AA) in B cell differentiation in vitro. AA-2G is stable in a solution unlike AA but is hydrolyzed by cellular alpha-glucosidase to release AA. Mouse spleen B cells were primed for 2 days with an anti-mu antibody in the presence of interleukin (IL)-4 and IL-5 and then washed and recultured with AA-2G in the presence of IL-4 and IL-5. AA-2G, but not AA, dose-dependently increased IgM production, the greatest enhancement being 150% at concentrations of more than 0.5 mM. In the absence of IL-4 and IL-5, primed B cells produced a negligible amount of IgM, and AA-2G had no effect. AA-2G-induced IgM production in the presence of IL-4 and IL-5 was inhibited by the alpha-glucosidase inhibitor castanospermine. Intracellular AA content, depleted during the priming period, increased by adding AA-2G at the start of reculture. Treatment of B cells with AA-2G resulted in an increase in the number of IgM-secreting cells, CD138-positive cells and CD45R/B220-negative cells. The number of viable cells in untreated cultures decreased gradually, but the decrease was significantly attenuated by AA-2G, resulting in about 70% more viable cells in AA-2G-treated cultures. AA-2G caused a slight but reproducible enhancement of DNA synthesis and a slight decrease in the number of cells with a sub-G1 DNA content. These results demonstrated that AA released from AA-2G enhanced cytokine-dependent IgM production in anti-mu-primed B cells and suggest that its effect is caused through promoting the differentiation of B cells to plasma cells and attenuating the gradual decrease in the number of viable cells

Filamentous structures in the cell envelope are associated with bacteroidetes gliding machinery

Many bacteria belonging to the phylum Bacteroidetes move on solid surfaces, called gliding motility. In our previous study with the Bacteroidetes gliding bacterium Flavobacterium johnsoniae, we proposed a helical loop track model, where adhesive SprB filaments are propelled along a helical loop on the cell surface. In this study, we observed the gliding cell rotating counterclockwise about its axis when viewed from the rear to the advancing direction of the cell and revealed that one labeled SprB focus sometimes overtook and passed another SprB focus that was moving in the same direction. Several electron microscopic analyses revealed the presence of a possible multi-rail structure underneath the outer membrane, which was associated with SprB filaments and contained GldJ protein. These results provide insights into the mechanism of Bacteroidetes gliding motility, in which the SprB filaments are propelled along tracks that may form a multi-rail system underneath the outer membrane. The insights may give clues as to how the SprB filaments get their driving force

Monkeys mutant for PKD1 recapitulate human autosomal dominant polycystic kidney disease.

Autosomal dominant polycystic kidney disease (ADPKD) caused by PKD1 mutations is one of the most common hereditary disorders. However, the key pathological processes underlying cyst development and exacerbation in pre-symptomatic stages remain unknown, because rodent models do not recapitulate critical disease phenotypes, including disease onset in heterozygotes. Here, using CRISPR/Cas9, we generate ADPKD models with PKD1 mutations in cynomolgus monkeys. As in humans and mice, near-complete PKD1 depletion induces severe cyst formation mainly in collecting ducts. Importantly, unlike in mice, PKD1 heterozygote monkeys exhibit cyst formation perinatally in distal tubules, possibly reflecting the initial pathology in humans. Many monkeys in these models survive after cyst formation, and cysts progress with age. Furthermore, we succeed in generating selective heterozygous mutations using allele-specific targeting. We propose that our models elucidate the onset and progression of ADPKD, which will serve as a critical basis for establishing new therapeutic strategies, including drug treatments

Identification of cell cycle–arrested quiescent osteoclast precursors in vivo

Osteoclasts are multinucleated cells that resorb bone. Although osteoclasts originate from the monocyte/macrophage lineage, osteoclast precursors are not well characterized in vivo. The relationship between proliferation and differentiation of osteoclast precursors is examined in this study using murine macrophage cultures treated with macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-κB (RANK) ligand (RANKL). Cell cycle–arrested quiescent osteoclast precursors (QuOPs) were identified as the committed osteoclast precursors in vitro. In vivo experiments show that QuOPs survive for several weeks and differentiate into osteoclasts in response to M-CSF and RANKL. Administration of 5-fluorouracil to mice induces myelosuppression, but QuOPs survive and differentiate into osteoclasts in response to an active vitamin D3 analogue given to those mice. Mononuclear cells expressing c-Fms and RANK but not Ki67 are detected along bone surfaces in the vicinity of osteoblasts in RANKL-deficient mice. These results suggest that QuOPs preexist at the site of osteoclastogenesis and that osteoblasts are important for maintenance of QuOPs