A single high‐resolution HLA mismatch has a similar adverse impact on the outcome of related hematopoietic stem cell transplantation as a single low‐resolution HLA mismatch

The relative importance of the resolution level of HLA typing has not been fully defined for related donor transplantation. To address this question, we retrospectively evaluated patients who underwent a first related hematopoietic stem cell transplantation (HSCT) from 2000 to 2011 from an HLA high‐resolution matched (MRD, n = 2,244), high‐resolution 1 locus‐mismatched (HR‐MMRD, n = 116), or low‐resolution 1 locus‐mismatched related donor (LR‐MMRD, n = 396) in the graft‐versus‐host direction at three loci (HLA A, B, and DRB1) using the database of the Japan Society for Hematopoietic Cell Transplantation. The median age was 40 years (0–74). The median follow‐up duration of surviving patients was 950 days. Although the cumulative incidences of grade III–IV acute graft‐versus‐host disease (GVHD) in the HR‐MMRD and LR‐MMRD groups were significantly higher than those in the MRD group (HR‐MMRD 19.8%, LR‐MMRD 20.4%, and MRD 9.5%), there was no statistically significant difference between the HR‐MMRD and LR‐MMRD groups (P = 0.65). Although both HR‐MMRD and LR‐MMRD were significantly associated with an increased risk of non‐relapse mortality and a worse overall survival, there was no statistically significant difference between the HR‐MMRD and LR‐MMRD groups. In conclusion, LR‐MM and HR‐MM have a similar adverse impact on the outcome in related HSCT. Am. J. Hematol. 90:618–623, 2015. © 2015 Wiley Periodicals, Inc.     

Feasibility of reduced-intensity conditioning followed by unrelated cord blood transplantation for primary hemophagocytic lymphohistiocytosis: a nationwide retrospective analysis in Japan

A nationwide retrospective analysis was performed on patients who received allogeneic hematopoietic stem cell transplantation for primary or familial hemophagocytic lymphohistiocytosis (HLH) in Japan. The present analysis investigated whether reduced-intensity conditioning (RIC) followed by cord blood transplantation (CBT) (RIC–CBT) is feasible, compared to the outcomes of myeloablative conditioning and bone marrow transplantation. Based on the JSHCT data, 53 patients were analyzed. The overall survival rate (OS) was 65.4 ± 6.6 %. RIC–CBT (n = 13) was not inferior to other methods. Patients with a performance status of PS 4 (ECOG scale) with HLH-associated severe organ dysfunction during the initiation of conditioning had extremely poor outcomes. The OS rate in the RIC–CBT patients, excluding those with a performance status 4, was 80.0 ± 12.6 %. RIC may reduce treatment-related mortality; in addition, patients with engraftment failure, which is the main adverse event following RIC–CBT, were successfully rescued with secondary CBT. Unrelated cord blood may represent an alternative source if a patient has no related donor. As a RIC regimen for CBT, 140 mg/m² melphalan with fludarabine and anti-lymphocyte globulin or anti-thymocyte globulin may be feasible, but further dosage optimization should be performed in controlled clinical trials.     

The SYNGAP1 3′UTR Variant in ALS Patients Causes Aberrant SYNGAP1 Splicing and Dendritic Spine Loss by Recruiting HNRNPK

Fused in sarcoma (FUS) is a pathogenic RNA-binding protein in amyotrophic lateral sclerosis (ALS). We previously reported that FUS stabilizes Synaptic Ras-GTPase activating protein 1 (Syngap1) mRNA at its 3′ untranslated region (UTR) and maintains spine maturation. To elucidate the pathologic roles of this mechanism in ALS patients, we identified the SYNGAP1 3′UTR variant rs149438267 in seven (four males and three females) out of 807 ALS patients at the FUS binding site from a multicenter cohort in Japan. Human-induced pluripotent stem cell (hiPSC)-derived motor neurons with the SYNGAP1 variant showed aberrant splicing, increased isoform α1 levels, and decreased isoform γ levels, which caused dendritic spine loss. Moreover, the SYNGAP1 variant excessively recruited FUS and heterogeneous nuclear ribonucleoprotein K (HNRNPK), and antisense oligonucleotides (ASOs) blocking HNRNPK altered aberrant splicing and ameliorated dendritic spine loss. These data suggest that excessive recruitment of RNA-binding proteins, especially HNRNPK, as well as changes in SYNGAP1 isoforms, are crucial for spine formation in motor neurons.SIGNIFICANCE STATEMENT It is not yet known which RNAs cause the pathogenesis of amyotrophic lateral sclerosis (ALS). We previously reported that Fused in sarcoma (FUS), a pathogenic RNA-binding protein in ALS, stabilizes synaptic Ras-GTPase activating protein 1 (Syngap1) mRNA at its 3′ untranslated region (UTR) and maintains dendritic spine maturation. To elucidate whether this mechanism is crucial for ALS, we identified the SYNGAP1 3′UTR variant rs149438267 at the FUS binding site. Human-induced pluripotent stem cell (hiPSC)-derived motor neurons with the SYNGAP1 variant showed aberrant splicing, which caused dendritic spine loss along with excessive recruitment of FUS and heterogeneous nuclear ribonucleoprotein K (HNRNPK). Our findings that dendritic spine loss is because of excess recruitment of RNA-binding proteins provide a basis for the future exploration of ALS-related RNA-binding proteins.     

Wear and surface roughness of current prosthetic composites after toothbrush/dentifrice abrasion

STATEMENT OF PROBLEM: Surface changes of prosthetic composites caused by toothbrushing are known, although composite materials have been improved and are now widely used for various kinds of prosthetic restorations. PURPOSE: This study evaluated the influence of toothbrushing on abrasive wear and surface roughness of current prosthetic composites. MATERIAL AND METHODS: Seven composite materials (Artglass, Axis, Cesead II, Conquest Sculpture, Estenia, Infis, and Targis) were assessed, and a machinable ceramic material (Cerec 2 Vitablocs) was used as a reference. Composite specimens polymerized with their proprietary curing units and sectioned ceramic specimens were stored in water for 14 days, and subsequently subjected to toothbrush-dentifrice abrasion. The amount of vertical loss and the surface roughness of each specimen after 20,000 strokes were determined with a profilometer. Average values of groups of 5 specimens were compared with ANOVA and Duncan new multiple range test. RESULT: Significantly (P < .05) less wear was observed with respect to the Targis (10.01 microm; SD = 0.53 microm) and Estenia (13.04 microm; 1.95 microm) materials than for the other composites assessed, whereas Artglass (34.08 microm; 3.66 microm) and Conquest Sculpture (31.78 microm; 4.67 microm) materials demonstrated the most wear. The least surface roughness was exhibited by Conquest Sculpture (Ra, 0.54 microm; 0.07 microm) material, and the greatest by Cesead II (1.10 microm; 0.13 microm). Ceramic material showed a more wear-resistant (4.54 microm; 0.79 microm) and smoother (0.26 microm; 0.02 microm) surface than any of the composite materials. CONCLUSION: Abrasion and surface roughness of the prosthetic composites caused by toothbrushing varied in accordance with the material. Type of prosthetic composite significantly influenced the surface condition after toothbrushing.     

Colour reproducibility of a photo-activated prosthetic composite with different thicknesses

This study determined the colour accuracy of a photo-activated prosthetic composite with various thicknesses and shades for the purpose of evaluating the relationship between material thickness variation and colour reproducibility. Four light shades (A1, B1, C1 and D2) and four dark shades (A4, B4, C4 and D4) of a representative photo-activated prosthetic composite (Artglass) for body paste were assessed. The specimens were prepared with thicknesses of 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mm. The L*a*b* colour parameter of each shade was measured using a dental colorimeter (ShadeEye) connected to a computer. The overall colour accuracy of groups of five specimens was compared by means of analysis of variance (ANOVA) and Scheffe's S multiple range test (P=0;05). In addition, L*a*b* colour difference (Delta E*) values between the 3.0 mm thickness and the other thicknesses were calculated. Three-factor ANOVA revealed that all colour parameters were influenced by the thickness as well as by the shade letter and shade tab (P < 0.05). The L* value consistently decreased for all shades as thickness increased, although there was no obvious correlation between the thickness of the material and either the a* or b* values. Among the Delta E* values, the values between the 2.5 and 3.0 mm thicknesses were the lowest, and values were found to be lower than 2.0 for all shades. The colour accuracy of the photo-activated prosthetic composite was related to the thickness of the material regardless of the shade. For acceptable colour reproducibility, a composite material thickness of at least 2.5 mm was found to be required.     

Hepatic mitochondrial cytochrome P-450: Isolation and functional characterization

A CO-binding heme protein was solubilized and partially purified from the inner membrane fraction of rat liver mitochondria by a modification of a method [Imai, Y. & Sato, R. (1974) Biochem. Biophys. Res. Commun. 60, 8-14] developed to purify cytochrome P-450 from liver microsomes. The partially purified preparation contained protoheme and its spectral properties are characteristic of the heme proteins of the cytochrome P-450 family. The isolated cytochrome P-450 preparation could reconstitute a CO-sensitive, NADPH-dependent 26-hydroxylation activity for 5beta-cholestane-3alpha,7alpha,-12alpha-triol when supplemented with NADPH-adrenodoxin reductase and adrenodoxin, both purified from bovine adrenocortical mitochondria. Unlike a cytochrome P-450 purified from liver microsomes of drug-untreated rats, however, the liver mitochondrial cytochrome P-450 could not catalyze NADPH-dependent benzphetamine N-demethylation in the presence of adrenodoxin reductase and adrenodoxin or function with the purified microsomal NADPH-cytochrome c reductase plus Emulgen 913 as an electron-donating system. It is concluded that the rat liver inner mitochondrial membrane houses a species of cytochrome P-450 functional in 5beta-cholestane-3alpha,7alpha,12alpha-triol 26-hydroxylation.     

Intestinal thrombotic microangiopathy after allogeneic bone marrow transplantation: a clinical imitator of acute enteric graft-versus-host disease

Thrombotic microangiopathy after bone marrow transplantation (post-BMT TMA) is a serious transplant-related complication. We identified 16 patients with TMA after allogeneic BMT who showed histopathological evidence of intestinal TMA in their gut specimens (six autopsies, 10 biopsies). In all, 14 patients had grade II-IV acute graft-versus-host disease (GVHD). The first seven patients were retrospectively diagnosed with TMA. Since six of them were diagnosed with progressive GVHD at that time because there was no awareness of the existence of intestinal TMA, they received more intensive treatment for GVHD, but all died between days +49 and +253. In contrast, the remaining nine patients were recently diagnosed with intestinal TMA on the basis of colonoscopic biopsies. For eight of these patients, the immunosuppressants were reduced, and the patients' intestinal symptoms improved gradually. Six of the nine patients were still alive 12 months after the diagnosis of TMA. Our findings suggest that the gut may be a site involved in post-BMT TMA, presenting as ischemic enterocolitis. Differentiating intestinal TMA from acute GVHD is important in patients suffering from severe and refractory diarrhea after BMT.