Background
Pre‐implantation genetic testing (PGT) has been performed worldwide since it was first used by Handyside et al in the United Kingdom to sex embryos in 1990. Until about 2010, cleavage ...stage embryo biopsy and fluorescent in situ hybridization (FISH) were mainstream; however, in 2012, blastocyst biopsy (trophectoderm; TE biopsy) became mainstream. In addition, array comparative genomic hybridization (aCGH) was used for analysis and further evolved to next‐generation sequencing (NGS), which is used worldwide.
Methods
PGT for reciprocal balanced translocation and Robertsonian translocation (PGT‐SR) was approved in Japan for habitual abortion to reduce pregnancy loss, and since 2008, we have been performing PGT‐SR using cleavage stage embryos and FISH. In 2014, we performed TE biopsy and NGS analysis.
Main findings
In this paper, I separately described the details of our methods and clinical results of FISH and NGS. NGS is superior to FISH because it can detect all chromosomes.
Conclusion
TE biopsy and NGS, which have recently become mainstream, have stable outcomes, because TE biopsy yields more cells and fewer mosaics than the cleavage stage. As a result, diagnoses are more reliable, resulting in higher pregnancy rates and lower abortion rates.
Clinical and industrial applications require large quantities of human induced pluripotent stem cells (hiPSCs); however, little is known regarding the mechanisms governing aggregate formation and ...stability in suspension culture. To address this, we determined differences in growth processes among hiPSC lines in suspension culture. Using an hiPSC aggregate suspension culture system, hiPSCs from different lines formed multicellular aggregates classified as large compact or small loose based on their size and morphology. Time-lapse observation of the growth processes of two different hiPSC lines revealed that the balance between cell division and the extent of subsequent cell death determined the final size and morphology of aggregates. Comparison of the cell survival and death of two hiPSC lines showed that the formation of small, loose aggregates was due to continued cell death during the exponential phase of growth, with apoptotic cells extruded from growing hiPSC aggregates by the concerted contraction of their neighbors. Western blot and immunofluorescent staining revealed that aggregate morphology and proliferative ability relied to a considerable extent upon secretion of the extracellular matrix (ECM). hiPSCs forming large compact and stable aggregates showed enhanced production of collagen type I in suspension culture at 120 h. Furthermore, these aggregates exhibited higher expression of E-cadherin and proliferation marker Ki-67 as compared with levels observed in small and loose aggregates at 120 h. These findings indicated that differences in both aggregate formation and stability in suspension culture among hiPSC lines were caused by differences in ECM secretion capacity.
Recently, the comprehensive concept of "infection-related glomerulonephritis (IRGN)" has replaced that of postinfectious glomerulonephritis (PIGN) because of the diverse infection patterns, ...epidemiology, clinical features, and pathogenesis. In addition to evidence of infection, hypocomplementemia particularly depresses serum complement 3 (C3), with endocapillary proliferative and exudative GN developing into membranoproliferative glomerulonephritis (MPGN); also, C3-dominant or co-dominant glomerular immunofluorescence staining is central for diagnosing IRGN. Moreover, nephritis-associated plasmin receptor (NAPlr), originally isolated from the cytoplasmic fraction of group A
, is vital as an essential inducer of C3-dominant glomerular injury and is a key diagnostic biomarker for IRGN. Meanwhile, "C3 glomerulopathy (C3G)", also showing a histological pattern of MPGN due to acquired or genetic dysregulation of the complement alternative pathway (AP), mimics C3-dominant IRGN. Initially, C3G was characterized by intensive "isolated C3" deposition on glomeruli. However, updated definitions allow for glomerular deposition of other complement factors or immunoglobulins if C3 positivity is dominant and at least two orders of magnitude greater than any other immunoreactant, which makes it challenging to quickly distinguish pathomorphological findings between IRGN and C3G. As for NAPlr, it was demonstrated to induce complement AP activation directly in vitro, and it aggravates glomerular injury in the development of IRGN. A recent report identified anti-factor B autoantibodies as a contributing factor for complement AP activation in pediatric patients with PIGN. Moreover, C3G with glomerular NAPlr deposition without evidence of infection was reported. Taken together, the clinico-pathogenic features of IRGN overlap considerably with those of C3G. In this review, similarities and differences between the two diseases are highlighted.
In several previous studies, Charlson comorbidity index (CCI) score was associated with postoperative complications, mortality, and re-admission. There are few reports about the influence of CCI ...score on postoperative clinical outcome. The purpose of this study was to investigate the influence of comorbidities as calculated with CCI on postoperative clinical outcomes after PLIF.
Three hundred sixty-six patients who underwent an elective primary single-level PLIF were included. Postoperative clinical outcome was evaluated with the Japanese Orthopaedic Association lumbar score (JOA score). The correlation coefficient between the CCI score and postoperative improvement in JOA score was investigated. Patients were divided into three groups according to their CCI score (0, 1, and 2+). JOA improvement rate, length of stay (LOS), and direct cost were compared between each group. Postoperative complications were also investigated.
There was a weak negative relationship between CCI score and JOA improvement rate (r = - 0.20). LOS and direct cost had almost no correlation with CCI score. The JOA improvement rate of group 0 and group 1 was significantly higher than group 2+. LOS and direct cost were also significantly different between group 0 and group 2+. There were 14 postoperative complications. Adverse postoperative complications were equivalently distributed in each group, and not associated with the number of comorbidities.
A higher CCI score leads to a poor postoperative outcome. The recovery rate of patients with two or more comorbidities was significantly higher than in patients without comorbidities. However, the CCI score did not influence LOS and increased direct costs. The surgeon must take into consideration the patient's comorbidities when planning a surgical intervention in order to achieve a good clinical outcome.
Novel deep learning image reconstruction (DLIR) reportedly changes the image quality characteristics based on object contrast and image noise. In clinical practice, computed tomography image noise is ...usually controlled by tube current modulation (TCM) to accommodate changes in object size. This study aimed to evaluate the image quality characteristics of DLIR for different object sizes when the in-plane noise was controlled by TCM. Images acquisition was performed on a GE Revolution CT system to investigate the impact of the DLIR algorithm compared to the standard reconstructions of filtered-back projection (FBP) and hybrid iterative reconstruction (hybrid-IR). The image quality assessment was performed using phantom images, and an observer study was conducted using clinical cases. The image quality assessment confirmed the excellent noise- reduction performance of DLIR, despite variations due to phantom size. Similarly, in the observer study, DLIR received high evaluations regardless of the body parts imaged. We evaluated a novel DLIR algorithm by replicating clinical behaviors. Consequently, DLIR exhibited higher image quality than those of FBP and hybrid-IR in both phantom and observer studies, albeit the value depended on the reconstruction strength, and proved itself capable of providing stable image quality in clinical use.
Expansive open-door laminoplasty results in favorable clinical outcomes for cervical myelopathy. However, some postoperative complications associated with surgical invasiveness, such as axial neck ...pain and kyphosis, have not been resolved. The use of an exoscope, which is a recently introduced novel magnification tool, allows for traditional open-door laminoplasty with minimal invasiveness. Therefore, we propose the use of exoscopic minimally invasive open-door laminoplasty (exLAP) and present its clinical outcomes during the acute postoperative period.
: A total of 28 patients who underwent open-door laminoplasty at C3-C6 were reviewed. Of these patients, 17 underwent exLAP (group M) and 11 underwent conventional Hirabayashi open-door laminoplasty (group H). Outcomes were evaluated using numerical rating scale (NRS) scores for neck pain and the frequency of oral analgesic use from postoperative day 1 to 7.
: The NRS score for neck pain was significantly lower for patients in group M than for those in group H.
: ExLAP is a novel, practical, and minimally invasive surgical technique that may alleviate the postoperative axial pain of patients with cervical myelopathy.
Ossification of the posterior longitudinal ligament of the spine (OPLL) is a common spinal disorder among the elderly that causes myelopathy and radiculopathy. To identify genetic factors for OPLL, ...we performed a genome-wide association study (GWAS) in ∼8,000 individuals followed by a replication study using an additional ∼7,000 individuals. We identified six susceptibility loci for OPLL: 20p12.3 (rs2423294: P = 1.10 × 10(-13)), 8q23.1 (rs374810: P = 1.88 × 10(-13)), 12p11.22 (rs1979679: P = 4.34 × 10(-12)), 12p12.2 (rs11045000: P = 2.95 × 10(-11)), 8q23.3 (rs13279799: P = 1.28 × 10(-10)) and 6p21.1 (rs927485: P = 9.40 × 10(-9)). Analyses of gene expression in and around the loci suggested that several genes are involved in OPLL etiology through membranous and/or endochondral ossification processes. Our results bring new insight to the etiology of OPLL.
Ossification of the posterior longitudinal ligament of the spine (OPLL) is an intractable disease leading to severe neurological deficits. Its etiology and pathogenesis are primarily unknown. The ...relationship between OPLL and comorbidities, especially type 2 diabetes (T2D) and high body mass index (BMI), has been the focus of attention; however, no trait has been proven to have a causal relationship. We conducted a meta-analysis of genome-wide association studies (GWASs) using 22,016 Japanese individuals and identified 14 significant loci, 8 of which were previously unreported. We then conducted a gene-based association analysis and a transcriptome-wide Mendelian randomization approach and identified three candidate genes for each. Partitioning heritability enrichment analyses observed significant enrichment of the polygenic signals in the active enhancers of the connective/bone cell group, especially H3K27ac in chondrogenic differentiation cells, as well as the immune/hematopoietic cell group. Single-cell RNA sequencing of Achilles tendon cells from a mouse Achilles tendon ossification model confirmed the expression of genes in GWAS and post-GWAS analyses in mesenchymal and immune cells. Genetic correlations with 96 complex traits showed positive correlations with T2D and BMI and a negative correlation with cerebral aneurysm. Mendelian randomization analysis demonstrated a significant causal effect of increased BMI and high bone mineral density on OPLL. We evaluated the clinical images in detail and classified OPLL into cervical, thoracic, and the other types. GWAS subanalyses identified subtype-specific signals. A polygenic risk score for BMI demonstrated that the effect of BMI was particularly strong in thoracic OPLL. Our study provides genetic insight into the etiology and pathogenesis of OPLL and is expected to serve as a basis for future treatment development.
Activated monocytes/macrophages promote glomerular injury, including crescent formation, in anti-glomerular basement membrane (GBM) glomerulonephritis. Disulfiram, an alcohol-aversion drug, inhibits ...monocyte/macrophage migration by inhibiting FROUNT, a cytosolic protein that enhances chemokine receptor signaling. Our study found that disulfiram at a human equivalent dose successfully blocked albuminuria and crescent formation with podocyte loss, and later stage kidney fibrotic lesions, in a rat model of anti-GBM glomerulonephritis. A disulfiram derivative, DSF-41, with more potent FROUNT inhibition activity, inhibited glomerulonephritis at a lower dose than disulfiram. Disulfiram markedly reduced the number of monocytes or macrophages at the early stage of glomerulonephritis and that of CD3+ and CD8+ lymphocytes at the established stage. Impaired pseudopodia formation was observed in the glomerular monocytes/macrophages of the disulfiram group; consistent with the in vitro observation that disulfiram blocked chemokine-dependent pseudopodia formation and chemotaxis of bone marrow-derived monocytes/macrophages. Furthermore, disulfiram suppressed macrophage activation as revealed by reduced expression of inflammatory cytokines and chemokines (TNF-α, CCL2, and CXCL9) and reduced CD86 and MHC class II expressions in monocytes/macrophages during glomerulonephritis. The dramatic reduction in monocyte/macrophage number might have resulted from disulfiram suppression of both the chemotactic response of monocytes/macrophages and their subsequent activation to produce cytokines and chemokines, which further recruit monocytes. Additionally, FROUNT was expressed in CD68+ monocytes/macrophages infiltrating the crescentic glomeruli in human anti-GBM glomerulonephritis. Thus, disulfiram can be a highly effective and safe drug for the treatment of glomerulonephritis by blocking the chemotactic responses of monocytes/macrophages and their activation status in the glomerulus.
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