Recruitment of DNA repair synthesis machinery to sites of DNA damage/repair in living human cells

The eukaryotic sliding DNA clamp, proliferating cell nuclear antigen (PCNA), is essential for DNA replication and repair synthesis. In order to load the ring-shaped, homotrimeric PCNA onto the DNA double helix, the ATPase activity of the replication factor C (RFC) clamp loader complex is required. Although the recruitment of PCNA by RFC to DNA replication sites has well been documented, our understanding of its recruitment during DNA repair synthesis is limited. In this study, we analyzed the accumulation of endogenous and fluorescent-tagged proteins for DNA repair synthesis at the sites of DNA damage produced locally by UVA-laser micro-irradiation in HeLa cells. Accumulation kinetics and in vitro pull-down assays of the large subunit of RFC (RFC140) revealed that there are two distinct modes of recruitment of RFC to DNA damage, a simultaneous accumulation of RFC140 and PCNA caused by interaction between PCNA and the extreme N-terminus of RFC140 and a much faster accumulation of RFC140 than PCNA at the damaged site. Furthermore, RFC140 knock-down experiments showed that PCNA can accumulate at DNA damage independently of RFC. These results suggest that immediate accumulation of RFC and PCNA at DNA damage is only partly interdependent

Pectoralis Major and Serratus Anterior Muscle Flap for Diaphragmatic Reconstruction

We have reported a new reconstruction method using a pectoralis major and serratus anterior muscle flap for diaphragmatic defects after chondrosarcoma resection. The reconstruction of diaphragmatic defects is challenging. In diaphragmatic reconstruction with chest wall defects, strong chest wall reconstruction and diaphragmatic flexibility are important to avoid interference with respiration. The artificial material Gore-Tex is used as the first choice, but it has infection-, exposure-, and durability-related drawbacks. As an alternative method using artificial material, we have reported our new technique—diaphragmatic reconstruction using a reversed-combined pectoralis major and serratus anterior muscle flap

Wireless data transmission in a 560-GHz band using low-phase-noise terahertz wave generated by photomixing of a pair of distributed feedback lasers injection-locking to Kerr micro-resonator soliton comb

The demand for higher data rates in next-generation mobile wireless communication systems (6G) has led to significant interest in terahertz (THz) waves as a high-frequency, broad modulation bandwidth carrier wave. In this study, we propose and demonstrate a wireless data transfer in the 560-GHz band using low-phase-noise THz waves generated by photomixing of a pair of distributed feedback lasers injection-locking to Kerr micro-resonator soliton comb. Experimental results showed near-error-free on-off keying (OOK) data transfer at 1 Gbit/s in the 560-GHz band, with a Q-factor of 6.23, surpassing the error-free limit. Also, modulation formats of binary phase shift keying (BPSK) and quadrature phase shift keying (QPSK) were successfully used, showing clear constellation diagrams and relatively low root mean squared error vector magnitude (rms EVM) values of 23.9% and 23.6%, respectively. Moreover, data transfer at 0.4 Gbit/s in 16 quadrature amplitude modulation (16QAM) demonstrated clear isolated symbols and achieved a low rms EVM value of 8.1%, complying with the IEEE 802.15.3d standard amendment. These demonstrations highlight the potential of using injection-locked DFB lasers with the Kerr micro-resonator soliton comb to achieve high-quality, high-speed wireless data transfer in the 560-GHz band. These findings contribute significantly to the advancement of wireless communication technology in the THz frequency range and pave the way for the realization of 6G wireless communication systems

The impact of crystal phase transition on the hardness and structure of kidney stones

The version of record of this article, first published in Urolithiasis, is available online at Publisher’s website: https://doi.org/10.1007/s00240-024-01556-5.Calcium oxalate kidney stones, the most prevalent type of kidney stones, undergo a multi-step process of crystal nucleation, growth, aggregation, and secondary transition. The secondary transition has been rather overlooked, and thus, the effects on the disease and the underlying mechanism remain unclear. Here, we show, by periodic micro-CT images of human kidney stones in an ex vivo incubation experiment, that the growth of porous aggregates of calcium oxalate dihydrate (COD) crystals triggers the hardening of the kidney stones that causes difficulty in lithotripsy of kidney stone disease in the secondary transition. This hardening was caused by the internal nucleation and growth of precise calcium oxalate monohydrate (COM) crystals from isolated urine in which the calcium oxalate concentrations decreased by the growth of COD in closed grain boundaries of COD aggregate kidney stones. Reducing the calcium oxalate concentrations in urine is regarded as a typical approach for avoiding the recurrence. However, our results revealed that the decrease of the concentrations in closed microenvironments conversely promotes the transition of the COD aggregates into hard COM aggregates. We anticipate that the suppression of the secondary transition has the potential to manage the deterioration of kidney stone disease

Evidence for Solution-Mediated Phase Transitions in Kidney Stones: Phase Transition Exacerbates Kidney Stone Disease

Maruyama M., Tanaka Y., Momma K., et al. Evidence for Solution-Mediated Phase Transitions in Kidney Stones: Phase Transition Exacerbates Kidney Stone Disease. Crystal Growth and Design 23, 4285 (2023); https://doi.org/10.1021/acs.cgd.3c00108.In this study, we investigated calcium oxalate (CaOx) kidney stones and showed direct evidence of the solution-mediated phase transition of calcium oxalate dihydrate (COD; the metastable phase) to calcium oxalate monohydrate (COM; the stable phase). We examined the crystal phases, crystal textures, and protein distributions within thin sections of calcium oxalate kidney stones. Observation with a polarized-light microscope showed that the outline of the mosaic texture, in which COM crystals are assembled in a mosaic pattern, roughly coincides with COD’s crystallographically stable face angles. Microfocus X-ray CT measurement captured the intermediate process of the phase transition, starting inside the COD single crystal and gradually transforming to COM crystals. In addition, the distribution of osteopontin and prothrombin fragment-1, common proteins contained in urine and visualized by multicolor fluorescence immunostaining, showed no apparent striations inside the COM single crystals with the mosaic texture, although the striation is apparent inside the COD single crystals. This is probably because the phase transition of mosaic-like COM occurred in a semiclosed system inside the COD single crystal, so the effect of periodic (day-night, seasonal, etc.) urinary protein concentration changes was small. On the other hand, striations were visible in concentrically laminated COM. This indicated that concentrically laminated COM formed in response to the changes in urinary protein concentrations. From the above, we conclude that the COD single crystals and the concentrically laminated COM seen in CaOx stones are primary structures, and the mosaic COM is a secondary structure that is a pseudomorph formed by the solution-mediated phase transition from COD single crystals

Identification of the sex-determining factor in the liverwort Marchantia polymorpha reveals unique evolution of sex chromosomes in a haploid system

半数体生物の性染色体上の性決定遺伝子を解明 --コケがもつ現生生物最古の起源の性染色体--. 京都大学プレスリリース. 2021-11-08.Sex determination is a central process for sexual reproduction and is often regulated by a sex determinant encoded on a sex chromosome. Rules that govern the evolution of sex chromosomes via specialization and degeneration following the evolution of a sex determinant have been well studied in diploid organisms. However, distinct predictions apply to sex chromosomes in organisms where sex is determined in the haploid phase of the life cycle: both sex chromosomes, female U and male V, are expected to maintain their gene functions, even though both are non-recombining. This is in contrast to the X-Y (or Z-W) asymmetry and Y (W) chromosome degeneration in XY (ZW) systems of diploids. Here, we provide evidence that sex chromosomes diverged early during the evolution of haploid liverworts and identify the sex determinant on the Marchantia polymorpha U chromosome. This gene, Feminizer, encodes a member of the plant-specific BASIC PENTACYSTEINE transcription factor family. It triggers female differentiation via regulation of the autosomal sex-determining locus of FEMALE GAMETOPHYTE MYB and SUPPRESSOR OF FEMINIZATION. Phylogenetic analyses of Feminizer and other sex chromosome genes indicate dimorphic sex chromosomes had already been established 430 mya in the ancestral liverwort. Feminizer also plays a role in reproductive induction that is shared with its gametolog on the V chromosome, suggesting an ancestral function, distinct from sex determination, was retained by the gametologs. This implies ancestral functions can be preserved after the acquisition of a sex determination mechanism during the evolution of a dominant haploid sex chromosome system

In vivo efficacy of KRP-109, a novel elastase inhibitor, in a murine model of severe pneumococcal pneumonia.

KRP-109 is a novel specific inhibitor of neutrophil elastase (NE). Various studies suggest that NE inhibitors reduce lung injury associated with systemic inflammatory response syndrome (SIRS). In this study, the efficacy of KRP-109 was examined using a murine model of severe pneumonia induced by Streptococcus pneumoniae (S. pneumoniae). Female mice (CBA/J, aged 5 weeks) were inoculated intranasally with penicillin-susceptible S. pneumoniae (ATCC49619 strain, 2.5 × 10(8) CFU/mouse). KRP-109 (30 or 50 mg/kg) or physiological saline as a control was administered intraperitoneally every 8 h beginning at 8 h after inoculation, and survival rate was evaluated over 7 days. Histopathological and bacteriological analyses of the lung, and bronchoalveolar lavage were performed at 48 h post-infection. The mice treated with KRP-109 (KRP-109 mice) tended to have higher survival rate than those given saline. The lung tissues of the KRP-109 mice had few neutrophils in the alveolar walls and less inflammation. Furthermore, KRP-109 decreased significantly total cell and neutrophil counts, and cytokine levels (interleukin 1β and macrophage inflammatory protein 2) in bronchoalveolar lavage fluid. Viable bacterial numbers in lung were not influenced by treatment of KRP-109. The present results indicate that KRP-109 reduces lung inflammation in a murine model, and that KRP-109 may be useful for the treatment of patients with severe pneumonia

糖尿病誘発性慢性腎疾患モデル動物―尿細管障害バイオマーカーによる検討―

Diabetes-induced chronic kidney disease（ DCKD） is a serious health problem. Therefore, modalities for preventing/improving DCKD are required. The aim of this study was to establish an animal model of DCKD. Four five-week-old male OLETF and LETO rats were used as experimental and control animals, respectively. After both rats were raised 17 months, collections of urine, blood and bilateral kidneys were performed. Urinary biomarkers for renal proximal tubule injury, monocyte chemoattractant protein-1（ MCP-1） and kidney injury molecule-1（ KIM-1）, were measured by the Bio-Plex system. OLETF rats showed a significant increase in blood glucose, Hb A1c, creatinine, blood urea nitrogen（ BUN）, kidney/body weight ratio, glomerular area, 24 h-urine volume, urinary protein concentrations, MCP-1 and KIM-1 compared with LETO rats. There was a positive correlation not only between BUN and MCP-1 or KIM-1 but also between urinary protein concentrations and MCP-1 or KIM-1. These results indicate that the present OLETF rats have both glomerular and proximal tubular injury. Thus, the aged male OLETF rats with long-term diabetes may become a potent animal model for examining effects of exercise/dietary therapies on DCKD