Biology subjects

Kazama, T.

Publications and source records attributed to Kazama, T..

8 recordsLinked to original sources

bioRxiv2025

Efficacy of vocal cord injection of dedifferentiated fat cells in treating glottis closure insufficiency: Insights from a rat model of recurrent laryngeal nerve resection

Glottic insufficiency results from impaired vocal cord contact, leading to a gap between the cords and manifesting as hoarseness and respiratory difficulties. Vocal cord injection is a commonly utilized therapeutic approach to rectify this gap by augmenting vocal cord volume; however, the optimal injectable material remains undetermined. Dedifferentiated fat cells (DFATs), derived from mature adipocytes, exhibit robust proliferative capacity and multipotency, establishing them as potential candidates for treating glottic insufficiency. This study investigated the therapeutic efficacy of DFATs in a rat model of recurrent laryngeal nerve paralysis. In the experimental design, the recurrent laryngeal nerve of each rat was resected unilaterally, and 5 weeks later, the atrophied vocal cord muscles were injected with either 10 l of saline (control), 1.0 x 106 DFATs in 10 l (DFAT group), or 1.0 x 106 DFATs combined with 500 ng of basic fibroblast growth factor (bFGF) in 10 l (DFAT + bFGF group). At 4 weeks post-injection, laryngeal endoscopy was performed to evaluate the glottic gap, followed by histological analyses to assess vocal muscle atrophy, collagen deposition, and markers of cellular proliferation (Ki-67) and apoptosis (TUNEL). To confirm DFAT engraftment, GFP-labeled DFATs were evaluated at 2, 4, and 6 weeks post-injection. Results indicated that both the DFAT and DFAT + bFGF groups exhibited significantly reduced glottic gaps, increased collagen deposition, and decreased TUNEL-positive apoptotic cells compared to controls. Notably, the DFAT + bFGF group displayed superior outcomes, including a greater vocal muscle area and enhanced Ki-67-positive cell proliferation, indicating mitigation of muscle atrophy. GFP-positive DFATs were detectable in the tissue for up to 6 weeks, confirming engraftment. These findings underscore the potential of DFATs, particularly in combination with bFGF, as an innovative and effective therapeutic approach for glottic insufficiency secondary to recurrent laryngeal nerve paralysis.

cell biology↗

bioRxiv2024

Engineering rice Nramp5 modifies cadmium and manganese uptake selectivity using yeast assay system

Cd is a seriously hazardous heavy metal for both plants and humans and international regulations regarding Cd intake have become stricter in recent years. Three-quarters of the Cd intake comes from plant-based foods, half of which comes from cereals. Therefore, it is anticipated that the Cd uptake efficiency of cereals, including rice, a staple crop in Asia, will be reduced. Natural resistance-associated macrophage protein (Nramp) is the principal transporter involved in the uptake and translocation of metal ions in various plants. In rice, OsNramp5 is a transporter of Mn, which is an essential micronutrient for plant growth, and is responsible for Cd uptake. Although several attempts have been made to engineer the metal uptake characteristics of OsNramp5, in many cases, both Cd and Mn uptake efficiencies are impaired. Therefore, in this study, we engineered OsNramp5 to reduce Cd uptake while retaining Mn uptake efficiency for low-Cd rice production. OsNramp5 was engineered using amino acid substitution(s) at the 232nd Ala and 235th Met of OsNramp5, which have been suggested to be key residues for metal uptake efficiency and/or selectivity by structural analyses of bacterial Nramps. The metal uptake efficiency was first analyzed using a yeast model assay system. Several mutants showed less than 8.6% Cd and more than 64.1% Mn uptake efficiency compared to the original OsNramp5. The improved metal uptake characteristics were confirmed by direct measurement of the metal content in the yeast using inductively coupled plasma optical emission spectroscopy. Notably, several mutants reduced Cd uptake efficiency to the background level while retaining more than 64.7% Mn uptake efficiency under conditions mimicking heavily polluted soils in the world. In addition, computational structural modeling suggested requirements for the spatial and chemical properties of the metal transport tunnel and metal-binding site, respectively, for Cd/Mn uptake efficiency.

plant biology↗

bioRxiv2023

Impact of climate change and oligotrophication on quality and quantity of lake primary production: A case study in Lake Biwa

Global climate change and anthropogenic oligotrophication are expected to reshape the dynamics of primary production (PP) in aquatic ecosystems; however, few studies have explored their long-term effects. In theory, the PP of phytoplankton in Lake Biwa may decline over decades due to warming, heightened stratification, and anthropogenic oligotrophication. Furthermore, the PP of large phytoplankton, which are inedible to zooplankton, along with biomass-specific productivity (PBc), could decrease. In this study, data from 1976-2021 and active fluorometry measurements taken in 2020 and 2021 were evaluated. Quantitatively, the temporal dynamics of mean seasonal PP during 1971-2021 were assessed according to the carbon fixation rate to investigate relationships among environmental factors. Qualitatively, phytoplankton biomass, PP, and PBc were measured in two size fractions [edible (S) or inedible (L) for zooplankton] in 2020 and 2021, and the L:S balance for these three measures was compared between 1992 (low-temperature/high-nutrient conditions) and 2020-2021 (high-temperature/low-nutrient conditions) to assess seasonal dynamics. The results indicated that climate change and anthropogenic oligotrophication over the past 50 years have diminished Lake Biwas PP since the 1990s, impacting the phenology of PP dynamics. However, the L:S balance in PP and PBc has exhibited minimal change since 1992. These findings suggest that, although climate change and oligotrophication may reduce overall PP, they do not markedly alter the inedible/edible phytoplankton balance in terms of PP and PBc. Instead, as total PP declines, the production of small edible phytoplankton decreases proportionally, potentially affecting trophic transfer efficiency and material cycling in Lake Biwa.

ecology↗

bioRxiv2023

Similarity and dissimilarity in alteration of gene expression profile associated with inhalational anesthesia between sevoflurane and desflurane

Although sevoflurane is the most commonly used inhalational anesthetic agent, the popularity of desflurane is increasing to a similar level. The main beneficial property of desflurane is the relatively fast emergence of the patient from the anesthetic state after halting its supply compared with anesthesia using other anesthetic agents. However, there has been no comprehensive comparison of the effects of these two anesthetic agents on alterations in liver gene expression profiles in animals, including humans, to assess the levels of hepatotoxicity that is induced at least in some extent by inhalational anesthesia. Thus, we compared alterations in the global gene expression profiles in the livers of rats subjected to inhalational anesthesia by sevoflurane or desflurane by a next-generation sequencing method. Our data revealed that both anesthetic agents significantly activated a similar set of genes including those related to drug metabolism and circadian rhythm. Furthermore, many genes downregulated by sevoflurane were also downregulated by desflurane. However, many of the genes related to the cholesterol biosynthetic process were specifically repressed by sevoflurane, but not by desflurane.

molecular biology↗

bioRxiv2022

The mitochondrial genome of Oryza sativa L. cv. Taichung 65 (Poaceae)

A complete mitochondrial genome sequence was determined by a hybrid approach with long- and short-read sequences in a japonica rice cultivar Taichung 65. The assembled mitochondrial genome was 401,060 bases in length and an overall GC content of 43.88% and predicted to harbor 57 protein-encoding, 18 transfer RNA, and 3 ribosomal RNA genes. Phylogenetic and sequence polymorphism analysis indicated that Taichung 65 is classified as typical japonica. The mitochondrial genome resource would serve as a fundamental tool to investigate cytoplasmic male sterility in rice.

genomics↗

bioRxiv2021

Size-dependent susceptibility of lake phytoplankton to light stress: An implication for succession of large green algae in a deep oligotrophic lake

Field observations of the population dynamics and measurements of photophysiology in Lake Biwa were conducted by size class (< vs. > 30 m) from early summer to autumn to investigate the relationships between susceptibility to light stress and cell size. Also, a nutrient bioassay was conducted to clarify whether the growth rate and photosystem II (PSII) photochemistry of small and large phytoplankton are limited by nutrient availability. Large phytoplankton, which have lower intracellular Chl-a concentrations, had higher maximum PSII photochemical efficiency (Fv/Fm) but lower non-photochemical quenching (NPQNSV) than small phytoplankton under both dark and increased light conditions. The nutrient bioassay revealed that the PSII photochemistry of small phytoplankton was restricted by N and P deficiency at the pelagic site even at the end of the stratification period, while that of large phytoplankton was not. These results suggest that large phytoplankton have lower susceptibility to PSII photodamage than small phytoplankton due to lower intracellular Chl-a concentrations. The size dependency of susceptibility to PSII photoinactivation may play a key role in large algal blooms in oligotrophic water.

microbiology↗

bioRxiv2021

Disruption of the mitochondrial gene orf352 partially restores pollen development in cytoplasmic male sterile rice

Plant mitochondrial genomes sometimes carry cytoplasmic male sterility (CMS)-associated genes. These genes have been harnessed in agriculture to produce high-yielding F1 hybrid seeds in various crops. The gene orf352 was reported to be an RT102-type CMS gene in rice (Oryza sativa), although a causal demonstration of its role in CMS is lacking. Here, we employed mitochondrion-targeted transcription activator-like effector nucleases (mitoTALENs), to knock out orf352 from the mitochondrial genome in the cytoplasmic male sterile rice RT102A. We isolated 18 independent transformation events in RT102A that resulted in genome editing of orf352, including its complete removal from the mitochondrial genome in several plants. Sequence analysis around the mitoTALEN target sites revealed the repair of their induced double-strand breaks via homologous recombination. Near the 5' target site, repair involved sequences identical to orf284, while repair of the 3' target site yielded various new sequences that generated new chimeric genes consisting orf352 fragments. Plants with a new mitochondrial gene encoding amino acids 179 to 352 of ORF352 exhibited the same shrunken pollen grain phenotype as RT102A, whereas plants either lacking orf352 or harboring a new gene encoding amino acids 211 to 352 of ORF352 showed partial rescue of pollen viability and germination, although they failed to set seed. These results demonstrated that disruption of orf352 partially restored pollen development, indicating that amino acids 179 to 210 from ORF352 may contribute to the establishment of pollen abortion.

plant biology↗

bioRxiv2020

Development of photosynthetic carbon fixation model using multi-excitation wavelength fast repetition rate fluorometry in Lake Biwa

Direct measurements of gross primary productivity (GPP) in the water column are essential, but can be spatially and temporally restrictive. Fast repetition rate fluorometry (FRRf) is a bio-optical technique based on chlorophyll a (Chl-a) fluorescence that can estimate the electron transport rate (ETRPSII) at photosystem II (PSII) of phytoplankton in real time. However, derivation of phytoplankton GPP in carbon units from ETRPSII remains challenging because the electron requirement for carbon fixation ([FE]e,C) can vary depending on multiple factors. Also, the FRRf is still relatively novel, especially in freshwater ecosystems where phosphorus limitation and cyanobacterial blooms are common. The goal of the present study is to construct a robust [FE]e,C model for freshwater ecosystems using simultaneous measurements of ETRPSII by FRRf with multi-excitation wavelengths coupled with traditional carbon fixation rate by the 13C method. The study was conducted in oligotrophic and mesotrophic areas in Lake Biwa from July 2018 to May 2019. The combination of excitation light at 444, 512 and 633 nm correctly estimated ETRPSII of cyanobacteria. The range of [FE]e,C in the phytoplankton community varied from 1.1 to 31.0 mol e- mol C-1 during the study period. Generalized liner model showed the best model including 12 physicochemical and biological factors explained 67% of the variance in [FE]e,C. Among all factors, water temperature was the most significant, while PAR intensity was not. The GPP values estimated by FRRf (GPPf) with the best [FE]e,C model relative to 13C (GPP13C) varied 0.5-1.5. Further, GPPf estimated with more parsimonious [FE]e,C models were also comparable to GPP13C. This study quantifies the applicability of the in situ FRRf methodology, and supports continuous monitoring of GPP by FRRf in lakes with large spatio-temporal variability of environmental conditions and phytoplankton assemblages.

microbiology↗