Percutaneous Transluminal Angioscopy During Coronary Intervention

To investigate the feasibility of angioscopic-guided percutaneous transluminal coronary angioplasty and to elucidate the mechanism of efficacy of coronary stenting for acute myocardial infarction, we performed coronary angioscopy in 102 patients with stable angina or acute myocardial infarction. Thrombi and intimal flaps were observed in most patients after coronary angioplasty. Large intimal splits were seen in one third of patients. Stents were inserted in 10 patients who were revealed to have a large flap or protruding split to the inner lumen. Thrombolytic agents were administered in 2 patients with large thrombi. Additional treatments were required in 32% of patients. No acute myocardial infarction or unstable angina occurred in patients during hospitalization. Thus, angioscopy of the coronary lumen enables clinicians to determine the most appropriate and least risky coronary intervention strategy. In patients with acute myocardial infarction, angioscopy revealed occlusive or protruding thrombi in 34 of 35 patients. The protruding thrombi disappeared after stenting. The frequency of large intimal flaps increased after predilatation with balloon, but these disappeared after stenting. The present angioscopic study demonstrates that the coronary stent compresses the occlusive or protruding thrombi and covers the ruptured thrombogenic plaque Consequently, smooth-surfaced and wide vessel lumen are obtained

Stromal interaction molecule 1 haploinsufficiency causes maladaptive response to pressure overload

Stromal interaction molecule 1 (STIM1), an endo/sarcoplasmic reticulum Ca2+ sensor, has been shown to control a Ca2+- dependent signal that promotes cardiac hypertrophy. However, whether STIM1 has adaptive role that helps to protect against cardiac overload stress remains unknown. We hypothesized that STIM1 deficiency causes a maladaptive response to pressure overload stress. We investigated STIM1 heterozygous KO (STIM1(+/)-) mice hearts, in which STIM1 protein levels decreased to 27% of wild-type (WT) with no compensatory increase in STIM2. Under stress-free conditions, no significant differences were observed in electrocardiographic and echocardiographic parameters or blood pressure between STIM1(+/)-and WT mice. However, when STIM1(+/)-mice were subjected to transverse aortic constriction (TAC), STIM1(+/-) mice had a higher mortality rate than WT mice. The TAC-induced increase in the heart weight to body weight ratio (mean mg/g +/- standard error of the mean) was significantly inhibited in STIM1(+/-) mice (WT sham, 4.12 +/- 0.14; WT TAC, 6.23 +/- 0.40; STIM1(+/-) sham, 4.53 +/- 0.16; STIM1(+/-) TAC, 4.63 +/- 0.08). Reverse transcription- polymerase chain reaction analysis of the left ventricles of TAC-treated STIM1(+/-) mice showed inhibited induction of cardiac fetal genes, including those encoding brain and atrial natriuretic proteins. Western blot analysis showed upregulated expression of transient receptor potential channel 1 (TRPC1) in TAC-treated WT mice, but suppressed expression in TAC-treated STIM1(+/-) mice. Taken together, the hearts of STIM1 haploinsufficient mice had a superficial resemblance to the WT phenotype under stress-free conditions; however, STIM1 haploinsufficient mice showed a maladaptive response to cardiac pressure overload

Correction of an asymmetric maxillary dental arch by alveolar bone distraction osteogenesis

This case report describes a new surgical orthodontic approach involving alveolar bone distraction osteogenesis for correction of an asymmetric maxillary dental arch. The treatment was combined with conventional orthognathic surgery to improve the mandibular lateral deviation. This new treatment strategy produced an ideal dental arch and a symmetric facial appearance efficiently and effectively

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target

A Comparative Study of The Internal Ribosome Entry Sitecontaining Expression System

We analyzed transient and stable eukaryotic gene expression using internal ribosome entry site （IRES） and green fluorescent protein （GFP）-S65T （IRES-GFP） expression systems. The IRES-containing vector enables simple construction of a plasmid and easy detection of transfected cells, as well as an estimation of the levels of the gene of interest. This system is useful for measuring changes in the intracellular calcium concentration using the fluorescent calcium indicator Fura-2 AM, although subcellular localization of the protein of interest is not detectable. Importantly, the IRES-GFP plasmid showed delayed expression and an approximate 40% reduction in fluorescence intensity, which suggests that usage of IRES-containing expression system might cause under-estimation of desired gene. Although this technique has some disadvantages, the IRES-containing vector is simple and straightforward for gene expression analysis, comparing with co-expression procedure of multiple plasmids

Diverse structures and adsorption properties of quasi-Werner-type copper(II) complexes with flexible and polar axial bonds

The quasi-Werner-type copper(II) complex, [Cu(PF6)2(4-mepy)4] (1), in which 4-mepy is the 4-methylpyridine ligand, has flexible and polar axial bonds of Cu-PF6. Flexibility of the Cu-PF6 bonds induces diverse and unprecedented guest-inclusion structures, such as {[Cu(PF6)2(4-mepy)4][Cu(PF6)(4-mepy)4(acetone)]・PF6・4acetone} (γ-1⊃2.5acetone), {[Cu(PF6)2(4-mepy)4][Cu(PF6)(4-mepy)4(2-butanone)]・PF6・3.5(2-butanone)} (γ-1⊃2.25(2-butanone)), {[Cu(PF6)2(4-mepy)4][Cu(PF6)(4-mepy)4(H2O)]・PF6・4benzene} (γ-1⊃0.5H2O・2benzene), and {[Cu(PF6)2(4-mepy)4]・2benzene} (γ-1⊃2benzene). Exposure of the dense form, α-1, to benzene vapor affords the benzene-inclusion complex {[Cu(PF6)2(4-mepy)4]・2benzene} (γ-1⊃2benzene), all benzene guests of which are easily removed by vacuum drying, reforming guest-free, dense α-1' with smaller sized crystals than α-1. In contrast to α-1, which shows almost no CO2 adsorption, α-1' adsorbs CO2 gas with structural transformations, this being the first example that exhibits adsorption of gas in a dense Werner-type complex and a drastic change in adsorption properties depending on the size of the crystals

A simple and dual expression plasmid system in prokaryotic (E. coli) and mammalian cells.

We introduce a simple and universal cloning plasmid system for gene expression in prokaryotic (Escherichia coli) and mammalian cells. This novel system has two expression modes: the (subcloning) prokaryotic and mammalian modes. This system streamlines the process of producing mammalian gene expression plasmids with desired genes. The plasmid (prokaryotic mode) has an efficient selection system for DNA insertion, multiple component genes with rare restriction sites at both ends (termed "units"), and a simple transformation to mammalian expression mode utilizing rare restriction enzymes and re-ligation (deletion step). The new plasmid contains the lac promoter and operator followed by a blunt-end EcoRV recognition site, and a DNA topoisomerase II toxin-originated gene for effective selection with isopropyl-β-D-thiogalactoside (IPTG) induction. This system is highly efficient for the subcloning of blunt-end fragments, including PCR products. After the insertion of the desired gene, protein encoded by the desired gene can be detected in E. coli with IPTG induction. Then, the lac promoter and operator are readily deleted with 8-nucleotide rare-cutter blunt-end enzymes (deletion step). Following re-ligation and transformation, the plasmid is ready for mammalian expression analysis (mammalian mode). This idea (conversion from prokaryotic to mammalian mode) can be widely adapted. The pgMAX system overwhelmingly simplifies prokaryotic and mammalian gene expression analyses