Evaluation of the tetrodotoxin uptake ability of pufferfish Takifugu rubripes tissues according to age using an in vitro tissue slice incubation method

The tetrodotoxin (TTX) uptake ability of pufferfish Takifugu rubripes tissues and its growth-associated changes were investigated using an in vitro tissue slice incubation method. Tissue slices prepared from the liver, skin, and intestine of a non-toxic cultured adult T. rubripes (20 months old) and incubated with incubation buffer containing 25 μg/mL TTX for 1?48 h showed a time-dependent increase in the TTX content in all tissues. The TTX contents of the skin and intestine slices were comparable to or slightly higher than that of the liver slices, with a similar transition pattern, suggesting similar TTX uptake ability among the skin, intestine, and liver. The TTX uptake ability of the liver and intestine did not differ significantly between young (8 months old) and adult (20 months old) fish, but the skin slices of young fish took up approximately twice as much TTX as that of adult fish, suggesting that the TTX uptake ability of the skin is involved in the growth-dependent changes in the toxin distribution inside the body in T. rubripes. To estimate the TTX uptake pathway in each tissue, an immunohistochemical technique was used to observe temporal changes in the intra-tissue microdistribution of TTX during incubation. The findings suggested that TTX is transferred and accumulates from pancreatic exocrine cells to hepatic parenchymal cells in the liver, from connective tissues to basal cells in the skin, and from villi epithelial cells via the lamina propria to the muscle layer in the intestine

Olfactory Response in Patients with Cerebral Palsy: Investigation of the Possible Use of Olfactory Testing to Evaluate Patients and Reduce Anxiety of Their Parents

Cerebral palsy (CP) is a developmental disorder associated with lifelong motor impairment and disability. Several studies have investigated the effects of CP on sensory responses, but there has been no research on olfactory function in CP. This study had two specific aims. First, by observing respiratory responses which serve as an index for the ability to detect and recognize odors, we aimed to estimate the patients\u27 ability to distinguish pleasant from unpleasant odors. These responses can also indicate the capacity for an emotional response to pleasant or unpleasant stimuli. Second, we aimed to investigate the psychological response in the parents as they observed the respiratory response in their children during the odor test. Parents were assessed with the Spielberger State-Trait Anxiety Inventory (STAI) to measure their state anxiety levels before and after they had observed their children being tested. In patients with CP, respiratory rate (RR) significantly decreased during pleasant odor stimuli (P < 0.05). There was a significant increase in RR during the presentation of the unpleasant odor (P < 0.05). All the CP patients showed respiratory changes in response to both pleasant and unpleasant smells, and we found that the parents\u27 anxiety levels decreased after the test. We intend to develop this olfactory test technique to contribute to the neurological rehabilitation for CP patients and to reduce anxiety of their parents

Evaluation of the tetrodotoxin uptake ability of pufferfish Takifugu rubripes tissues according to age using an in vitro tissue slice incubation method

The tetrodotoxin (TTX) uptake ability of pufferfish Takifugu rubripes tissues and its growth-associated changes were investigated using an in vitro tissue slice incubation method. Tissue slices prepared from the liver, skin, and intestine of a non-toxic cultured adult T. rubripes (20 months old) and incubated with incubation buffer containing 25 μg/mL TTX for 1–48 h showed a time-dependent increase in the TTX content in all tissues. The TTX contents of the skin and intestine slices were comparable to or slightly higher than that of the liver slices, with a similar transition pattern, suggesting similar TTX uptake ability among the skin, intestine, and liver. The TTX uptake ability of the liver and intestine did not differ significantly between young (8 months old) and adult (20 months old) fish, but the skin slices of young fish took up approximately twice as much TTX as that of adult fish, suggesting that the TTX uptake ability of the skin is involved in the growth-dependent changes in the toxin distribution inside the body in T. rubripes. To estimate the TTX uptake pathway in each tissue, an immunohistochemical technique was used to observe temporal changes in the intra-tissue microdistribution of TTX during incubation. The findings suggested that TTX is transferred and accumulates from pancreatic exocrine cells to hepatic parenchymal cells in the liver, from connective tissues to basal cells in the skin, and from villi epithelial cells via the lamina propria to the muscle layer in the intestine

The Influence of Experience on Gazing Patterns during Endovascular Treatment: Eye-Tracking Study

Objective: In various fields, differences in eye-gazing patterns during tasks between experts and novices have been evaluated. The aim of this study was to investigate gazing patterns during neuro-endovascular treatment using an eye-tracking device and assess whether gazing patterns depend on the physician’s experience or skill. Methods: Seven physicians performed coil embolization for a cerebral aneurysm in a silicone vessel model under biplane X-ray fluoroscopy, and their gazing patterns were recorded using an eye-tracking device. The subjects were divided into three groups according to experience, highly experienced (Expert) group, intermediately experienced (Trainee) group, and less experienced (Novice) group. The duration of fixation on the anterior–posterior (AP) view screen, lateral (LR) view, and out-of-screen were compared between each group. Results: During microcatheter navigation, the Expert and Trainee groups spent a long time on fixation to AP, while the Novice group split their attention between each location. In coil insertion, the Expert group gazed at both the AP and the LR views with more saccades between screens. In contrast, the Trainee group spent most of their time only on the AP view screen and the Novice group spent longer out-of-screen. Conclusion: An eye-tracking device can detect different gazing patterns among physicians with several experiences and skill levels of neuroendovascular treatment. Learning the gazing patterns of experts using eye tracking may be a good educational tool for novices and trainees

Production of Single-Chain Fv Antibodies Specific for GA-Pyridine, an Advanced Glycation End-Product (AGE), with Reduced Inter-Domain Motion

Due to their lower production cost compared with monoclonal antibodies, single-chain variable fragments (scFvs) have potential for use in several applications, such as for diagnosis and treatment of a range of diseases, and as sensor elements. However, the usefulness of scFvs is limited by inhomogeneity through the formation of dimers, trimers, and larger oligomers. The scFv protein is assumed to be in equilibrium between the closed and open states formed by assembly or disassembly of VH and VL domains. Therefore, the production of an scFv with equilibrium biased to the closed state would be critical to overcome the problem in inhomogeneity of scFv for industrial or therapeutic applications. In this study, we obtained scFv clones stable against GA-pyridine, an advanced glycation end-product (AGE), by using a combination of a phage display system and random mutagenesis. Executing the bio-panning at 37 °C markedly improved the stability of scFvs. We further evaluated the radius of gyration by small-angle X-ray scattering (SAXS), obtained compact clones, and also visualized ope

Multi-omics analysis defines highly refractory RAS burdened immature subgroup of infant acute lymphoblastic leukemia.

Funder: Princess Takamatsu Cancer Research FundFunder: Japan Foundation for Pediatric Research Funai Foundation for Information TechnologyKMT2A-rearranged infant acute lymphoblastic leukemia (ALL) represents the most refractory type of childhood leukemia. To uncover the molecular heterogeneity of this disease, we perform RNA sequencing, methylation array analysis, whole exome and targeted deep sequencing on 84 infants with KMT2A-rearranged leukemia. Our multi-omics clustering followed by single-sample and single-cell inference of hematopoietic differentiation establishes five robust integrative clusters (ICs) with different master transcription factors, fusion partners and corresponding stages of B-lymphopoietic and early hemato-endothelial development: IRX-type differentiated (IC1), IRX-type undifferentiated (IC2), HOXA-type MLLT1 (IC3), HOXA-type MLLT3 (IC4), and HOXA-type AFF1 (IC5). Importantly, our deep mutational analysis reveals that the number of RAS pathway mutations predicts prognosis and that the most refractory subgroup of IC2 possesses 100% frequency and the heaviest burden of RAS pathway mutations. Our findings highlight the previously under-appreciated intra- and inter-patient heterogeneity of KMT2A-rearranged infant ALL and provide a rationale for the future development of genomics-guided risk stratification and individualized therapy