Outcomes of Patients with Pulmonary Atresia with Intact Ventricular Septum Reaching Adulthood

Background: There is limited information on outcomes of adult patients with pulmonary atresia with intact ventricular septum (PA-IVS) due to the low incidence of disease and the large variation of surgical histories. Methods: Among 58 patients with repaired PA-IVS, a total of 32 patients aged ≥16 years and who were followed at our institution between January 2003 and December 2018 were reviewed. Surgical history, clinical outcomes, and laboratory, echocardiographic and electrocardiographic data were obtained by chart review. Results: Follow-up was from the age of 16 years and the median age at the latest follow-up was 23.7 years. Twenty-four patients had undergone biventricular repair (BVR), 3 had undergone one-and-a half ventricular repair (1.5VR), and 5 had undergone univentricular repair. Over a median follow-up period of 7.7 years (interquartile range: 4.1–11.0 years), 1 BVR patient died suddenly and 7 patients had heart failure. Arrhythmias were present in 5 patients. Ten patients underwent surgical re-interventions, including 4 BVR take-downs with conversion to 1.5VR and 3 Fontan conversions. Overall survival, heart failure-free, arrhythmia-free, and surgical re-intervention-free rates at 5 years and 10 years from the age of 16 years were 96.2% (95% confidence interval [CI], 77.2–99.4) and 96.2% (95% CI, 77.2– 99.4), 81.4% (95% CI, 62.1–92.1) and 74.6% (95%CI, 52.3–88.7), 88.7% (95% CI, 70.1–96.3) and 75.9% (95% CI, 51.7–90.2), and 80.7% (95% CI, 60.8–91.8) and 70.8% (95% CI, 49.7–85.7), respectively. Conclusion: Adults with PA-IVS have preserved long-term survival regardless of the early operative strategy, while they are at risk for heart failure, arrhythmia, and surgical re-intervention. Thus, detailed and continued follow-up is mandatory for all PA-IVS patients from childhood to adulthood

iPSC-derived type IV collagen α5-expressing kidney organoids model Alport syndrome

ヒトiPS細胞から作製した腎オルガノイドを用いたアルポート症候群病態モデルの開発. 京都大学プレスリリース. 2023-09-28.iPSC-derived kidney organoids to model a lifelong renal disease. 京都大学プレスリリース. 2023-10/17.Alport syndrome (AS) is a hereditary glomerulonephritis caused by COL4A3, COL4A4 or COL4A5 gene mutations and characterized by abnormalities of glomerular basement membranes (GBMs). Due to a lack of curative treatments, the condition proceeds to end-stage renal disease even in adolescents. Hampering drug discovery is the absence of effective in vitro methods for testing the restoration of normal GBMs. Here, we aimed to develop kidney organoid models from AS patient iPSCs for this purpose. We established iPSC-derived collagen α5(IV)-expressing kidney organoids and confirmed that kidney organoids from COL4A5 mutation-corrected iPSCs restore collagen α5(IV) protein expression. Importantly, our model recapitulates the differences in collagen composition between iPSC-derived kidney organoids from mild and severe AS cases. Furthermore, we demonstrate that a chemical chaperone, 4-phenyl butyric acid, has the potential to correct GBM abnormalities in kidney organoids showing mild AS phenotypes. This iPSC-derived kidney organoid model will contribute to drug discovery for AS

“Transmantle sign”を示す限局性皮質異形成における神経細胞の成熟と分化の未熟性：層特異的マーカー発現による解析

Transmantle dysplasia is a rare type of focal cortical dysplasia (FCD) characterized by expansion of the cortex from the deep white matter to the surface and in which there is a FCD IIA or IIB pathologic pattern. To characterize possible mechanisms underlying this regional disorder of radial migrating cells, we studied the expression patterns of neocortical layer-specific markers using immunohistochemistry in surgical specimens from 5 FCD IIA and 4 FCD IIB cases in children. All neuronal cells expressed the mature neuron marker MAP2/2B but not the microglia markers Iba-1 and CD68. Some layer-specific markers showed distinct expression patterns. TBR1-positive, SATB2-positive, and FOXP1-positive cells were diffusely distributed in the cortex and/or the white matter. TBR1-positive and FOXP1-positive cells were generally more numerous in FCD IIB than in FCD IIA and were mostly in the cortical molecular and upper layers. FOXP1-, FOXP2-, and CUTL1-positive cells also expressed the immature neuron marker, Nestin/PROX1, whereas TBR1-, CTIP2-, and SATB2-positive cells only expressed MAP2/2B. These data highlight differences between FCD IIB and FCD IIA with more cells having the immature marker in upper layer markers in the former. By analyzing layer-specific marker expression patterns, we identified apparent neuronal maturation differences between FCD IIA and FCD IIB in cases of transmantle dysplasia.博士（医学）・乙第1312号・平成25年5月29

A model relating transpiration for Japanese cedar and cypress plantations with stand structure

Previous studies have revealed that changes in forest structure due to management (e.g., thinning, aging, and clearcutting) could affect the forest water balance. However, there are unexplained variability in changes in the annual water balance with changing structure among different sites. This is the case even when analyzing data for specific species/regions. For a more advanced and process-based understanding of changes in the water balance with changing forest structure, we examined transpiration (E) observed using the sap-flux method for 14 Japanese cedar and cypress plantations with various structure (e.g., stem density and diameter) in Japan and surrounding areas and developed a model relating E with structural parameters. We expressed E using the simplified Penman–Monteith equation and modeled canopy conductance (G[c]) as a product of reference G[c] (G[cref]) when vapor pressure deficit is 1.0 kPa and functions expressing the responses of G[c] to meteorological factors. We determined G[cref] and parameters of the functions for the sites separately. E observed for the 14 sites was not reproduced well by the model when using mean values of G[cref] and the parameters among the sites. However, E observed for the sites was reproduced well when using G[cref] determined for each site and mean values of the parameters of the functions among the sites, similar to the case when using G[cref] and the parameters of the functions determined for each site. These results suggest that considering variations in G[cref] among the sites was important to reproduce variations in E, but considering variations in the parameters of the functions was not. Our analysis revealed that G[cref] linearly related with the sapwood area on a stand scale (A) and that Alinearly related with stem density (N) and powers of the mean stem diameter (d[m]). Thus, we proposed a model relating E with A (or N and d[m]), where G[cref] was calculated from A (or N and d[m]) and the parameters of the functions were assumed to be the mean values among the sites. This model estimates changes in Ewith changing structure from commonly available data (N and d[m]), and therefore helps improve our understanding of the underlying processes of the changes in the water balance for Japanese cedar and cypress plantations

Efficient integration of transgenes into a defined locus in human embryonic stem cells

Random integration is one of the more straightforward methods to introduce a transgene into human embryonic stem (ES) cells. However, random integration may result in transgene silencing and altered cell phenotype due to insertional mutagenesis in undefined gene regions. Moreover, reliability of data may be compromised by differences in transgene integration sites when comparing multiple transgenic cell lines. To address these issues, we developed a genetic manipulation strategy based on homologous recombination and Cre recombinase-mediated site-specific integration. First, we performed gene targeting of the hypoxanthine phosphoribosyltransferase 1 (HPRT) locus of the human ES cell line KhES-1. Next, a gene-replacement system was created so that a circular vector specifically integrates into the targeted HPRT locus via Cre recombinase activity. We demonstrate the application of this strategy through the creation of a tetracycline-inducible reporter system at the HPRT locus. We show that reporter gene expression was responsive to doxycycline and that the resulting transgenic human ES cells retain their self-renewal capacity and pluripotency

Widely Targeted Metabolomics Based on Large-Scale MS/MS Data for Elucidating Metabolite Accumulation Patterns in Plants

Metabolomics is an ‘omics’ approach that aims to analyze all metabolites in a biological sample comprehensively. The detailed metabolite profiling of thousands of plant samples has great potential for directly elucidating plant metabolic processes. However, both a comprehensive analysis and a high throughput are difficult to achieve at the same time due to the wide diversity of metabolites in plants. Here, we have established a novel and practical metabolomics methodology for quantifying hundreds of targeted metabolites in a high-throughput manner. Multiple reaction monitoring (MRM) using tandem quadrupole mass spectrometry (TQMS), which monitors both the specific precursor ions and product ions of each metabolite, is a standard technique in targeted metabolomics, as it enables high sensitivity, reproducibility and a broad dynamic range. In this study, we optimized the MRM conditions for specific compounds by performing automated flow injection analyses with TQMS. Based on a total of 61,920 spectra for 860 authentic compounds, the MRM conditions of 497 compounds were successfully optimized. These were applied to high-throughput automated analysis of biological samples using TQMS coupled with ultra performance liquid chromatography (UPLC). By this analysis, approximately 100 metabolites were quantified in each of 14 plant accessions from Brassicaceae, Gramineae and Fabaceae. A hierarchical cluster analysis based on the metabolite accumulation patterns clearly showed differences among the plant families, and family-specific metabolites could be predicted using a batch-learning self-organizing map analysis. Thus, the automated widely targeted metabolomics approach established here should pave the way for large-scale metabolite profiling and comparative metabolomics

Effects of soil water decline on diurnal and seasonal variations in sap flux density for differently aged Japanese cypress (Chamaecyparis obtusa) trees

The effects of soil drought on transpiration are often neglected when predicting transpiration for forests in humid regions under the influence of the Asian monsoon. These effects have indeed been neglected for Japanese cypress, Chamaecyparis obtusa, a major plantation species in Japan and the surrounding area, probably because previous studies have reported no clear effects of soil drought on transpiration for Japanese cypress forests. However, a few studies have reported an apparent reduction in transpiration with soil drought for young Japanese cypress forests. It remains unclear whether such a reduction in transpiration is limited to young Japanese cypress forests or if it is not uncommon for mature Japanese cypress forests, which occupy a large area in Japan. To clarify this point, we conducted sap flux measurements in a year with soil drought on three differently aged Japanese cypress stands including mature (43 years old) and relatively young (23 and 26 years old) trees. In a diurnal time scale, a cross correlation analysis of sap flux density (Fd) and vapor pressure deficit (VPD) showed that the time lags between Fd and VPD were 1-3 h in dry soil conditions. These were larger than those of wet soil conditions (&lt;1 h) for all sample trees. Fd at a given VPD in dry soil conditions was smaller than that in wet soil conditions for all sample trees; a 28%–63% reduction in the rate of change in Fd was observed under dry soil conditions. Because our results were obtained when the non-exceedance probability of recorded monthly precipitation was 9%–18%, the results suggest the need to consider the effects of soil drought more extensively. Those effects should be considered for not only relatively young but also mature Japanese cypress when predicting diurnal and seasonal patterns of transpiration in years with soil drought, and when predicting inter-annual patterns of transpiration for Japanese cypress despite humid temperate climate. </p