Search

Complications after hematopoietic stem cell transplantation (HSCT) especially graft versus host disease (GVHD) are serious events and the keys to success of HSCT. Pathological diagnosis of complications is critical but sometimes not definite in GVHD diagnosis. In this review, we focus on the role of macrophages in HSCT complications. In the early period after HSCT, residual recipient macrophages have important roles for engraftment and rejection. Hemophagocytosis is the main feature of activated recipient macrophages. Skin lesions after HSCT are usually skin GVHD but mimicking histology of interface dermatitis is hardly differentiated. Macrophages and lymphocytes of these lesions were studied and increased numbers of macrophages were significantly associated with overall survival of HSCT. It could be suggested macrophages were good predictive markers for skin GVHD‐like lesions. Intestinal type transplantation‐associated microangiopathy (i‐TAM) is an independent pathogenesis from GVHD with serious prognosis. Activated macrophages in these lesions are important and key to the pathogenesis of i‐TAM. These activated macrophages are predominantly residual recipient tissue‐resident macrophages. Cryptogenic organizing pneumonitis is a lung complication around day 100 after HSCT caused by activated alveolar macrophages. They are donor‐derived tissue‐resident macrophages. Residual recipient and donor‐derived macrophages work in different ways in HSCT complications. [ABSTRACT FROM AUTHOR]

Aims: Refractory cytopenia of childhood (RCC) is subdivided into myelodysplastic syndrome with multilineage dysplasia (MDS‐MLD) and RCC without (w/o) multilineage dysplasia (RCC without MLD). Although RCC is a histomorphological distinct entity, the bone marrow (BM) histology of RCC is not yet characterised in relation to multilineage dysplasia. We investigated the BM histological features of RCC to clarify the characteristics of BM histology of MDS‐MLD in childhood compared to RCC without MLD. Methods and results: The BM histology and cytology in 60 RCC patients from the nationwide registry of Japanese Childhood AA‐MDS Study Group were reviewed retrospectively. Although a thorough genetic assessment, including GATA2 and/or SAMD9, was not performed, inherited BM failure disorders were excluded by a cytogenetic test, a chromosome fragility test and a telomere length measurement along with careful clinical assessments. Among the 60 patients, 20 (33%) of MDS‐MLD and 40 (67%) of RCC w/o MLD were classified according to their BM cytology. We then investigated the BM histological features and compared them between the two groups. The BM cellularity, distribution pattern of haematopoiesis, frequency of left‐shifted granulopoiesis, numbers of micromegakaryocytes and p53 immunostaining‐positive cells were significantly different between the groups. The BM histology of MDS‐MLD in childhood showed higher cellularity, the more common occurrence of diffuse distribution pattern, more frequently left‐shifted granulopoiesis and more micromegakaryocytes and p53 immunostaining‐positive cells than RCC without MLD. Conclusions: Our results showed that MDS‐MLD in childhood had a characteristic BM histology compared to RCC without MLD. The clinical relevance of MDS‐MLD in childhood needs to be evaluated. [ABSTRACT FROM AUTHOR]

We review new trends of agricultural applications of atmospheric‐pressure plasmas for decontaminating agricultural products and enhancing seed germination and growth of plants and beneficial microorganisms. We classify the decontamination techniques into three kinds of treatments, which are gas‐phase, in‐liquid, and plasma‐activated liquid treatments, and introduce recent studies mainly after 2010, discussing the inactivation mechanism of microorganisms in each treatment. Likewise, we categorize the enhancement techniques into the same three kinds of treatments, and introduce the growth enhancements in detail, discussing the mechanisms and future prospects. [ABSTRACT FROM AUTHOR]

Inhalation of molecular hydrogen (H2) gas ameliorates oxidative stress-induced acute injuries in the brain. Consumption of water nearly saturated with H2 also prevents chronic neurodegenerative diseases including Parkinson’s disease in animal and clinical studies. However, the molecular mechanisms underlying the remarkable effect of a small amount of H2 remain unclear. Here, we investigated the effect of H2 on mitochondria in cultured human neuroblastoma SH-SY5Y cells. H2 increased the mitochondrial membrane potential and the cellular ATP level, which were accompanied by a decrease in the reduced glutathione level and an increase in the superoxide level. Pretreatment with H2 suppressed H2O2-induced cell death, whereas post-treatment did not. Increases in the expression of anti-oxidative enzymes underlying the Nrf2 pathway in H2-treated cells indicated that mild stress caused by H2 induced increased resistance to exacerbated oxidative stress. We propose that H2 functions both as a radical scavenger and a mitohormetic effector against oxidative stress in cells. [ABSTRACT FROM AUTHOR]

TP53 mutation is associated with various hematological malignancies and immunohistochemistry of p53 has been used as a simple method to establish the presence of a TP53 mutation. Since the significance of p53 expression is controversial in myelodysplastic syndrome (MDS) patients treated with azacitidine (Aza), we analyzed the prevalence of p53 expression as a prognostic factor in 60 MDS patients treated with Aza. To assess p53 expression, immunohistochemical analyses of bone marrow clot sections were performed. Overall survival (OS) was significantly lower in p53-positive patients compared with p53-negetive patients (59% vs. 85% at 12 months; P = 0.006). Multivariate analysis demonstrated that p53-positive was a significant prognostic factor for OS along with poor cytogenetics. Here, we propose a new prognostic index to make short-term prognoses of MDS patients in the era of Aza treatment; high: p53-positive and poor cytogenetics, low: p53-negative and absence of poor cytogenetics, and intermediate: the others. OS was significantly different among the three groups according to this index (Low 92%, Intermediate 65% and High 27% at 12 months; P < 0.0001). In conclusion, p53 expression was a significant prognostic factor in MDS patients treated with Aza. In combination with cytogenetic abnormalities, it is possible to make short-term prognoses. [ABSTRACT FROM AUTHOR]

Pyruvate therapy is a promising approach for the treatment of mitochondrial diseases. To identify novel biomarkers for diagnosis and to evaluate therapeutic efficacy, we performed microarray analysis of 2SD cybrid cells harboring a MELAS-causing mutation and control cells treated with either lactate or pyruvate. We found that expression and secretion of growth differentiation factor 15 (GDF15) were increased in 2SD cells treated with lactate and that serum GDF15 levels were significantly higher in patients with mitochondrial diseases than in those with other diseases, suggesting that GDF15 could be a useful marker for diagnosis and evaluating the therapeutic efficacy of pyruvate. [ABSTRACT FROM AUTHOR]

Low-temperature plasma (LTP) science and technology are rapidly establishing their place in agriculture, with already existing or envisaged future commercial applications in (i) sustainable fertilizer production, (ii) food safety "from farm to fork", and (iii) in reducing pollution and pathogens. On-demand local fertilizer production or pathogen inactivation at the farm by LTP technology will reduce the need for complex logistics. This current S.I. consists of one review article and 14 original research papers, focusing on state-of-the-art LTP applications in agricultural and food-related fields, ranging from the basics of activating air and water by electrical discharges and the treatment and decontamination of seeds and plants, to LTP use as a food processing technology. [Extracted from the article]

Inhalation of hydrogen (H2) gas is therapeutically effective for cerebrovascular diseases, neurodegenerative disorders, and neonatal brain disorders including pathologies induced by anesthetic gases. To understand the mechanisms underlying the protective effects of H2 on the brain, we investigated the molecular signals affected by H2 in sevoflurane‐induced neuronal cell death. We confirmed that neural progenitor cells are susceptible to sevoflurane and undergo apoptosis in the retrosplenial cortex of neonatal mice. Co‐administration of 1–8% H2 gas for 3 h to sevoflurane‐exposed pups suppressed elevated caspase‐3‐mediated apoptotic cell death and concomitantly decreased c‐Jun phosphorylation and activation of the c‐Jun pathway, all of which are induced by oxidative stress. Anesthesia‐induced increases in lipid peroxidation and oxidative DNA damage were alleviated by H2 inhalation. Phosphoproteome analysis revealed enriched clusters of differentially phosphorylated proteins in the sevoflurane‐exposed neonatal brain that included proteins involved in neuronal development and synaptic signaling. H2 inhalation modified cellular transport pathways that depend on hyperphosphorylated proteins including microtubule‐associated protein family. These modifications may be involved in the protective mechanisms of H2 against sevoflurane‐induced neuronal cell death. [ABSTRACT FROM AUTHOR]

Introduction: Frailty is a crucial health issue among older adults. Growth differentiation factor 15 (GDF15) is associated with inflammation, oxidative stress, insulin resistance, and mitochondrial dysfunction, which are possible pathogeneses of frailty. However, few longitudinal studies have investigated the association between GDF15 and the incidence of frailty. Therefore, we investigated whether high serum GDF15 levels are associated with the incidence of frailty. Methods: A total of 175 older adults (mean age: 77 ± 6 years; 63% women) with cardiometabolic diseases and no frailty out of the two criteria at baseline participated. Individuals with severe renal impairment or severe cognitive impairment were excluded. Serum GDF15 levels were measured at baseline. Patients were asked to assess frailty status at baseline and annually during follow-up using the modified version of the Cardiovascular Health Study (mCHS) and the Kihon Checklist (KCL). We examined the association between GDF15 tertiles and each frailty measure during follow-up (median 38–39 months). In the multivariate Cox regression analysis, with the GDF15 tertile groups as the explanatory variables, hazard ratios (HRs) and 95% confidence intervals (CIs) for incident frailty were calculated after adjusting for covariates and using the lowest tertile group as the reference. Results: During the follow-up period, 25.6% and 34.0% of patients developed frailty, as defined by the mCHS and KCL, respectively. The highest GDF15 tertile group had a significantly higher incidence of mCHS- or KCL-defined frailty than the lowest GDF15 tertile group. Multivariate Cox regression analysis revealed that the adjusted HRs for incident mCHS- and KCL-defined frailty in the highest GDF15 tertile group were 3.9 (95% CI: 1.3–12.0) and 2.7 (95% CI: 1.1–6.9), respectively. Conclusion: High serum GDF15 levels predicted the incidence of frailty among older adults with cardiometabolic diseases and could be an effective marker of the risk for frailty in interventions aimed at preventing frailty, such as exercise and nutrition. [ABSTRACT FROM AUTHOR]

Mitochondria are multifunctional organelles that regulate diverse cellular processes. Mitochondrial stress, including stress generated by electron transport chain defects and impaired mitochondrial proteostasis, is intimately involved in various diseases and pathological conditions. Sepsis is a life-threatening condition that occurs when an imbalanced host response to infection leads to organ dysfunction. Metabolic disturbances and impaired immune responses are implicated in the pathogenesis and development of sepsis. Given that mitochondria play central roles in cellular metabolism, mitochondrial stress is predicted to be involved in the pathological mechanism of sepsis. Under mitochondrial stress, cells activate stress response systems to maintain homeostasis. This mitochondrial stress response transcriptionally activates genes involved in cell survival and death. Mitochondrial stress also induces the release of distinctive secretory proteins from cells. Recently, we showed that growth differentiation factor 15 (GDF15) is a major secretory protein induced by mitochondrial dysfunction. In this article, we provide a brief overview of mitochondrial stress response and GDF15, and discuss the potential role of GDF15 in the pathophysiology of sepsis. • Mitochondrial stress response is partly mediated by the ISR pathway. • GDF15 is induced by mitochondrial dysfunction through the ISR pathway. • Septic patients and animals present mitochondrial dysfunction in multiple organs. • Circulating GDF15 level is elevated in patients and animals with sepsis. • GDF15 plays an important role in the pathophysiology of sepsis. [ABSTRACT FROM AUTHOR]

Micronodular thymoma with lymphoid stroma (MNT) is a rare thymic epithelial neoplasm subtype characterized by a micronodular tumor cell growth pattern and abundant lymphoid stroma. Micronodular thymic carcinoma with lymphoid stroma (MNCA) is considered as a malignant counterpart of MNT and exhibits a growth pattern similar to that of MNT but has histologic features reminiscent of thymic squamous cell carcinoma, such as cytologic atypia and CD5 and CD117 immunoexpression. Although both MNT and MNCA are characterized by abundant lymphoid stroma, it remains unknown whether there are differences in infiltrating lymphocytes between MNT and MNCA. We analyzed the immune microenvironment profile in eight MNT and three MNCA cases. The cell density of CD8‐positive T cells was significantly higher in MNT than in MNCA, whereas that of FOXP3‐positive T cells was significantly higher in MNCA than in MNT. There was no significant difference in the cell density of programmed death protein 1‐positive T cells and programmed death ligand 1 expression between the MNT and MNCA cases. Our findings indicated that the immune microenvironment of MNCA differed from that of MNT and, compared with the T‐cell profile of MNT, that of MNCA was more suppressive to patients′ antitumor immune response. [ABSTRACT FROM AUTHOR]

This Special Issue highlights the topics from the PLASMA Conference 2017, which was held in Himeji, Japan, on November 20–24, 2017. This conference was jointly organized by the Japan Society of Applied Physics, The Japan Society of Plasma and Nuclear Fusion Research, The Physical Society of Japan, and the Japan Society for the Promotion of Science 153 Committee (Plasma Science for Materials). [ABSTRACT FROM AUTHOR]

Molecular hydrogen (H2) is an antioxidant and anti-inflammatory agent; however, the molecular mechanisms underlying its biological effects are largely unknown. Similar to other gaseous molecules such as inhalation anesthetics, H2 is more soluble in lipids than in water. A recent study demonstrated that H2 reduces radical polymerization-induced cellular damage by suppressing fatty acid peroxidation and membrane permeability. Thus, we sought to examine the effects of short exposure to H2 on lipid composition and associated physiological changes in SH-SY5Y neuroblastoma cells. We analyzed cells by liquid chromatography-high-resolution mass spectrometry to define changes in lipid components. Lipid class analysis of cells exposed to H2 for 1 hour revealed transient increases in glycerophospholipids including phosphatidylethanolamine, phosphatidylinositol, and cardiolipin. Metabolomic analysis also showed that H2 exposure for 1 hour transiently suppressed overall energy metabolism accompanied by a decrease in glutathione. We further observed alterations to endosomal morphology by staining with specific antibodies. Endosomal transport of cholera toxin B to recycling endosomes localized around the Golgi body was delayed in H2-exposed cells. We speculate that H2-induced modification of lipid composition depresses energy production and endosomal transport concomitant with enhancement of oxidative stress, which transiently stimulates stress response pathways to protect cells. [ABSTRACT FROM AUTHOR]

Purpose: Less-invasive early diagnosis of lung cancer is essential for improving patient survival rates. The purpose of this study is to demonstrate that serum comprehensive miRNA profile is high sensitive biomarker to early-stage lung cancer in direct comparison to the conventional blood biomarker using next-generation sequencing (NGS) technology combined with automated machine learning (AutoML). Methods: We first evaluated the reproducibility of our measurement system using Pearson's correlation coefficients between samples derived from a single pooled RNA sample. To generate comprehensive miRNA profile, we performed NGS analysis of miRNAs in 262 serum samples. Among the discovery set (57 patients with lung cancer and 57 healthy controls), 1123 miRNA-based diagnostic models for lung cancer detection were constructed and screened using AutoML technology. The diagnostic faculty of the best performance model was evaluated by inspecting the validation samples (74 patients with lung cancer and 74 healthy controls). Results: The Pearson's correlation coefficients between samples derived from the pooled RNA sample ≥ 0.98. In the validation analysis, the best model showed a high AUC score (0.98) and a high sensitivity for early stage lung cancer (85.7%, n = 28). Furthermore, in comparison to carcinoembryonic antigen (CEA), a conventional blood biomarker for adenocarcinoma, the miRNA-based model showed higher sensitivity for early-stage lung adenocarcinoma (CEA, 27.8%, n = 18; miRNA-based model, 77.8%, n = 18). Conclusion: The miRNA-based diagnostic model showed a high sensitivity for lung cancer, including early-stage disease. Our study provides the experimental evidence that serum comprehensive miRNA profile can be a highly sensitive blood biomarker for early-stage lung cancer. [ABSTRACT FROM AUTHOR]

Purpose: Intermittent claudication (IC) refers to leg pain that is induced by walking and relieved by rest. Neurogenic IC is usually associated with lumbar canal stenosis (LCS). We present rare findings from an autopsied patient who had neurogenic IC caused by vasculitis in the cauda equina. Methods: We performed antemortem neurological and electrophysiological assessments, sural nerve biopsy, and post-mortem examination of the spinal cord and brain. Results: A 61-year-old man noted sudden-onset leg pain that was not associated with any traumatic trigger. His leg pain consistently appeared when the patient walked and quickly faded on stopping. Spine surgery and cardiovascular departments both made a diagnosis of IC. However, magnetic resonance imaging (MRI) did not show LCS, and all ankle-brachial pressure indices were normal. He subsequently developed diffuse muscle weakness of the legs a month after disease onset. Myeloperoxidase antineutrophil cytoplasmic autoantibody was seropositive (140 IU/mL), and a sural nerve biopsy revealed axonal injury and angiitis. MRI showed multiple cerebral infarctions. He was diagnosed with microscopic polyangiitis (MPA) and underwent corticosteroid therapy. He died from complications two months after the onset. A post-mortem study revealed vasculitis in the subarachnoid space of the cauda equina, spinal cord, and brain parenchyma. The cauda equina showed a combined loss of small and large axonal fibres. The lumbar cord displayed central chromatolysis of the lower motor neurons. Conclusion: MPA is a rare cause of neurogenic IC when the symptom is acute and multimodal. Small-vessel vasculitis affecting the cauda equina may underlie MPA-associated IC. [ABSTRACT FROM AUTHOR]

Background: Acquired erythrocytosis can be classified into polycythemia vera (PV) and non‐neoplastic erythrocytosis (NNE). The vast majority of PV patients harbor JAK2 mutations, but differentiating JAK2 mutation‐negative PV from NNE is challenging due to a lack of definitive molecular markers. Methods: We studied the clinical features of 121 patients with erythrocytosis of which 47 (38.8%) were JAK2 mutation‐positive and also fulfilled the diagnostic criteria for PV, and 67 (55.4%) JAK2 mutation‐negative erythrocytosis patients who were diagnosed as NNE. Diagnosis was strictly based on driver mutation analysis and central pathology review. Results: No JAK2 mutation‐negative PV patients were found in our cohort. The NNE group showed significantly younger (p < 0.01) age with higher frequency of smoking (p < 0.001), alcohol consumption (p < 0.001), and diabetes mellitus (p < 0.05), whereas the PV group (n = 47) showed significantly higher white blood cell count, platelet count, and lactate dehydrogenase (p < 0.001). Although serum erythropoietin (EPO) levels were significantly higher in NNE compared to PV (p < 0.001), approximately 40% of the NNE patients had EPO levels below the lower range of normal, fulfilling a minor diagnostic criterion of PV and raising the possibility of PV misdiagnosis. Conclusion: Low EPO levels in JAK2 mutation‐negative erythrocytosis may not be a reliable diagnostic criterion for distinguishing PV from NNE. [ABSTRACT FROM AUTHOR]

Mitochondria are dysfunctional in post-senescent cells. Therefore, age-dependent impairment of mitochondrial energy production accompanied by excessive mitochondrial reactive oxygen species (ROS) is proposed to be a key driver of cellular senescence, which is a state of irreversible cell cycle arrest. However, it remains to be clarified whether mitochondrial dysfunction initiates or accelerates replicative senescence. In this study, we observed no increase in mitochondrial ROS or decrease in mitochondrial respiratory function in human TIG-1 fibroblasts in the transition phase, during which the population doubling rate gradually decreases due to the development of replicative senescence. The integrated stress response and expression of growth differentiation factor 15, which are triggered by respiratory chain deficiency, were also not induced in the transition phase. Mitochondria were elongated without aberrant cristae structures in the transition phase. Mitophagy-related protein levels started to decrease in the transition phase, but autophagic flux slightly increased during replicative senescence. These results suggest that mitochondrial dysfunction and excessive mitochondrial ROS generation do not occur predominately in the transition phase and may not play a role in the development of replicative senescence in normal diploid TIG-1 fibroblasts. • Mitochondrial dysfunction does not occur in the transition phase before senescence. • Excess ROS generation does not occur in the transition phase before senescence. • GDF15 expression through ISR is not induced during replicative senescence. • Mitochondria are elongated without aberrant cristae structures in senescent cells. • Mitophagy-related proteins decline during replicative senescence. [ABSTRACT FROM AUTHOR]

Neurolymphomatosis is a neurological manifestation of lymphoma that involves the cranial or spinal peripheral nerves, nerve roots, and plexus with direct invasion of neoplastic cells. Neurolymphomatosis is rare among patients with low‐grade lymphoma. We report an autopsied case of neurolymphomatosis that arose from follicular lymphoma. A 49‐year‐old woman who presented with pain of her neck and shoulder and numbness of her chin. Computed tomography revealed enlarged lymph nodes in her whole body, and biopsy from the axillary lymph node revealed grade 2 follicular lymphoma. Although the patient underwent chemotherapy, she gradually developed muscle weakness in the upper limbs and sensory disturbances of the trunk and limbs. 18F‐fluorodeoxyglucose positron emission tomography (FDG‐PET) revealed increased tracer uptake of the cervical nerve roots. Repeated FDG‐PET after additional therapy revealed progression of disease within the nerve roots and brachial plexus, whereas gadolinium‐contrast magnetic resonance imaging (MRI) showed weak enhancement of the cervical nerve roots without formation of mass lesions. She died after a total disease duration of 12 months. Postmortem observations revealed invasion of lymphoma cells into the cervical nerve roots, dorsal root ganglia, and subarachnoid spaces of the spinal cord. Neurolymphomatosis was prominent at the segments of C6–Th2. Combined loss of axons and myelin sheaths was observed in the cervical nerve roots and posterior columns. Lymphoma cells also invaded the cranial nerves. The subarachnoid and perivascular spaces of the brain demonstrated focal invasion of the lymphoma. Mass lesions were not observed in the central nervous system. The lymphoma cells did not show histological transformation to higher grades, and the density of the centroblasts remained at grade 2. Our report clarifies that low‐grade follicular lymphoma can manifest as neurolymphomatosis and central nervous system invasion in the absence of transformation toward higher histological grades. FDG‐PET may be more sensitive to non‐mass‐forming lesions, including neurolymphomatosis, than gadolinium‐contrast MRI. [ABSTRACT FROM AUTHOR]

Aim: Wood creosote (WC), an over‐the‐counter (OTC) drug, has been used as an anti‐diarrheal agent for many years; however, its safety in combination with other drugs has not been adequately investigated. The current study was conducted to evaluate whether WC may cause drug interactions through the inhibition of cytochrome P450 (CYP). Methods: The inhibitory effects of WC on ethoxyresorufin O‐deethylation, paclitaxel 6α‐hydroxylation, S‐warfarin 7‐hydroxylation, S‐mephenytoin 4′‐hydroxylation, bufuralol1′‐hydroxylation, and midazolam (MDZ) 1′‐hydroxylation were investigated using human liver microsomes as the marker activities for CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4, respectively. In addition, the effects of single and three‐day multiple treatments of WC (30 mg/kg) on CYP2C‐mediated tolbutamide (TB, 20 mg/kg) and CYP3A‐mediated MDZ (15 mg/kg) metabolism were investigated in rats in vivo. Results: WC at clinical doses did not show a considerable inhibitory effect on any of the CYP activities examined in the in vitro study. CYP2C9 and CYP2C19 activities were inhibited by high concentrations of the WC (100 μg/mL); however, the IC50 values were approximately 70 times higher than the reported circulating levels of the main ingredients in WC. Furthermore, there were no marked changes in the pharmacokinetic parameters of TB and MDZ (area under the serum concentration–time curve, maximum serum concentrations, and elimination half‐life) after single or three‐day multiple treatments with WC. Conclusion: These results suggest that clinical doses of WC are unlikely to cause drug interactions mediated by the inhibition of CYPs. [ABSTRACT FROM AUTHOR]

Summary: In 2008, the World Health Organization proposed a new entity of childhood myelodysplastic syndrome (MDS), which was referred to as refractory cytopenia of childhood (RCC). However, whether this morphological classification reflects clinical outcomes remains unclear. We performed a prospective evaluation of bone marrow morphology in 252 children with acquired bone marrow failure between 2009 and 2013. Of 252 patients, 63 were diagnosed with aplastic anaemia (AA), 131 with RCC without multilineage dysplasia (RCC‐w/o‐MLD) and 58 with RCC with MLD (RCC‐MLD). One patient with AA, three with RCC‐w/o‐MLD and nine with RCC‐MLD presented with chromosomal abnormalities at diagnosis (P = 0·001). The response rates to immunosuppressive therapy (IST) at 6 months and the cumulative incidence of clonal evolution at 5 years did not significantly differ among the three groups. A multivariate analysis revealed that the morphological classification of RCC‐MLD was a significant risk factor for secondary graft failure after haematopoietic cell transplantation (HCT) (P = 0·003). In view of these findings, RCC could be divided into two categories, RCC‐w/o‐MLD and RCC‐MLD, because children with this condition exhibited a distinct morphology, frequent chromosomal abnormalities at diagnosis and a high frequency of secondary graft failure after HCT. [ABSTRACT FROM AUTHOR]

Perforin secreted from cytotoxic lymphocytes plays a critical role in cancer immunosurveillance. The aim of this study was to investigate the therapeutic potential of liposomes containing perforin expression vector driven by the promotor of prostate-specific antigen (PSA). The anti-tumor effect of perforin was analyzed using prostate cancer (PC) PC-3 cells in which perforin expression was controlled by Tet-on system (PC-3PRF cells). Liposomes encapsulating PSA promoter-driven perforin expression vector (pLipo) were constructed for its specific expression in PC. The anti-tumor effect of pLipo was evaluated in vitro using docetaxel-resistant PC 22Rv1 PC cell line, 22Rv1DR, and PC-3 cells in the presence of human peripheral blood mono nuclear cells (PBMCs) and also in vivo using male nude mice bearing 22Rv1DR cell-derived tumor xenograft. Induction of perforin significantly inhibited growth of PC-3PRF cells. Treatment with pLipo induced perforin expression in 22Rv1DR cells expressing PSA but not in PC-3 cells lacking it. Treatment with pLipo at a low concentration was prone to inhibit growth of both cell lines and significantly inhibited growth of 22Rv1DR cells when co-incubated with PBMCs. The combined use of pLipo at a high concentration with PBMCs showed nearly complete inhibition of 22Rv1DR cell growth. Intravenous administration of pLipo via tail vein increased the level of perforin in tumor and serum and significantly decreased the tumor volume. Our results suggest that liposome-mediated PC-specific expression of perforin could be a novel therapy for advanced PC. [ABSTRACT FROM AUTHOR]

The WBC count (C), Hb value (D), platelet count (E), and LDH level (F) for the patients classified based on driver mutation status are shown. Finally, in the Results, under the subheading 'Noncanonical JAK2 and MPL mutant exhibits wild-type-equivalent levels of STAT5 activation', "One such patient was found to harbor MPL S204F in the abovementioned screening of noncanonical JAK2 and MPL mutations." now reads "One such patient was found to harbor MPL S204F." NC-JAK2/MPL: noncanonical JAK2 and MPL mutation. [Extracted from the article]

Finally, in the Results, under the subheading 'Noncanonical JAK2 and MPL mutant exhibits wild-type-equivalent levels of STAT5 activation', "One such patient was found to harbor MPL S204F in the abovementioned screening of noncanonical JAK2 and MPL mutations." now reads "One such patient was found to harbor MPL S204F." Comparison of TPO (C) and IL-6 (D) concentrations in the serum among the patients with TN-ET, patients with ET harboring driver mutations, and healthy controls. Graph: Figure 1 Comparison of clinical parameters between the TN-ET patients and mutated ET patients. [Extracted from the article]

A subset of essential thrombocythemia (ET) cases are negative for disease-defining mutations on JAK2, MPL, and CALR and defined as triple negative (TN). The lack of recurrent mutations in TN-ET patients makes its pathogenesis ambiguous. Here, we screened 483 patients with suspected ET in a single institution, centrally reviewed bone marrow specimens, and identified 23 TN-ET patients. Analysis of clinical records revealed that TN-ET patients were mostly young female, without a history of thrombosis or progression to secondary myelofibrosis and leukemia. Sequencing analysis and human androgen receptor assays revealed that the majority of TN-ET patients exhibited polyclonal hematopoiesis, suggesting a possibility of reactive thrombocytosis in TN-ET. However, the serum levels of thrombopoietin (TPO) and interleukin-6 in TN-ET patients were not significantly different from those in ET patients with canonical mutations and healthy individuals. Rather, CD34-positive cells from TN-ET patients showed a capacity to form megakaryocytic colonies, even in the absence of TPO. No signs of thrombocytosis were observed before TN-ET development, denying the possibility of hereditary thrombocytosis in TN-ET. Overall, these findings indicate that TN-ET is a distinctive disease entity associated with polyclonal hematopoiesis and is paradoxically caused by hematopoietic stem cells harboring a capacity for cell-autonomous megakaryopoiesis. [ABSTRACT FROM AUTHOR]

We aimed to develop a sandwich ELISA to detect prostate-specific membrane antigen (PSMA) on small extracellular vesicles (EVs) using T-cell immunoglobulin domain and mucin domain-containing protein 4 (Tim4) as a capture molecule for EVs and to evaluate its diagnostic potential in urologic malignancies. First, we optimized the conditions for sandwich ELISA measuring the PSMA level on EVs captured from serum by Tim4 and found that the use of highly-purified EVs released from Tim4 that had captured EVs in serum reduced the background. Second, we confirmed its validity by studying mouse xenograft model for prostate cancer (PC). Lastly, we measured PSMA-EVs in serum of patients with urologic malignancies. The PSMA-EV levels were significantly higher in metastatic PC and castration-resistant PC (CRPC) patients than in therapy-naïve PC patients. In renal cell carcinoma (RCC) patients, PSMA-EVs were elevated in those with metastasis compared with those without metastasis, which may reflect the development of the neovasculature positive for PSMA in tumors. In conclusion, we developed a sandwich ELISA for detection of PSMA-EVs using highly-purified EVs isolated from serum by Tim4. Our results suggest that PSMA-EVs may be useful to diagnose and monitor not only PC but also RCC and possibly other hypervascular solid tumors. [ABSTRACT FROM AUTHOR]

To identify the key neutral species from atmospheric‐pressure plasma for promoting the proliferation of fibroblast cells, an atmospheric‐pressure radical generator was employed. A cell viability assay showed that the proliferation of fibroblasts was significantly promoted through radical treatment, resulting in the highest promotion ratio of 24%. The densities of the neutral radicals supplied by the radical generator were measured via mass spectrometry at atmospheric pressure. To determine an effective factor for the promotion, the dose dependencies of these radicals on the promotion ratio were investigated. The promotion ratio exhibited a strong correlation only with the NO• dose. These results confirmed that NO• is a direct trigger of plasma‐enhanced cell proliferation. [ABSTRACT FROM AUTHOR]

The differential diagnoses included inflammatory myofibroblastic sarcoma, undifferentiated pleomorphic sarcoma, inflammatory leiomyosarcoma, Epstein-Barr virus (EBV)-positive inflammatory follicular dendritic cell sarcoma, malignant gastrointestinal stromal tumour, and sarcomatoid carcinoma. HS has a poor prognosis, with most patients dying within 2 years.1,2 Our patient received TKI therapy, and has remained healthy without HS or CML recurrence for 3 years. Histiocytic sarcoma (HS) is a rare myeloid-derived neoplasm with histiocytic differentiation and an aggressive clinical course.1 Recent studies have suggested a common clonal origin for various haematolymphoid disorders.2 However, HS coexisting with chronic myeloid leukaemia (CML) without tyrosine kinase inhibitors (TKIs) has not been reported.3 We report a case of CML, and an intestinal mass caused by HS, that showed a I BCR::ABL1 i translocation. [Extracted from the article]

To investigate the potential role of the hydroxyl radical (•OH) in cold atmospheric plasma (CAP) jet treatment, two fluorescence-based methodologies are utilised to measure DNA strand breaks. The first comprises a model system of a double-stranded DNA oligomer, where the respective strand ends are tagged with fluorophore and quencher molecules; and the second, a cell culture system reporting DNA strand breaks using the γ-H2AX assay. During the various CAP jet treatments, optical emission spectroscopy is used to detect the •OH in the gas phase and electron spin resonance is used to detect the •OH in solution. The CAP jet production of the •OH is shown to correlate to CAP jet induced DNA damage both with the DNA model and in biological cells. Results indicate that the CAP jet induces a higher degree of DNA damage when the CAP plume is in contact with the target solution. The potential of a 'plasma screen' based upon a hydrogel film, as a method to remove the DNA-damaging •OH species from reaching skin cells, is shown to significantly reduce DNA damage whilst facilitating the delivery of hydrogen peroxide. These findings could aid in the development of CAP jet-based applications where DNA damage is the objective (e.g. in cancer treatment) and others where it is to be avoided, e.g. in open-wound treatment and dermatology. [ABSTRACT FROM AUTHOR]

Background: There has been no clinically useful diagnostic or prognostic biomarker for renal cell carcinoma (RCC). Serum γ-glutamyltransferase (GGT) activity has been reported to be a prognostic marker for several types of cancer including RCC. Exosomes or small extracellular vesicles present in body fluids have potential as a biomarker. We have recently demonstrated that GGT activity on exosomes isolated from serum is useful for the differential diagnosis of prostate cancer and benign prostate hyperplasia. In this study, we aimed to examine if serum exosomal GGT activity could be a marker for RCC. Methods: We examined GGT1 expression and GGT activity in cell lysates and exosomes from culture medium of HK-2 proximal tubule epithelial and RCC cell lines. GGT activity was measured using a fluorescent probe for GGT, γ-glutamyl hydroxymethyl rhodamine green. Serum and serum exosomal GGT activities were measured in patients with RCC. GGT1 expression in RCC tissues was evaluated by immunohistochemical staining. Results: GGT1 levels in exosomes from KMRC-1, OS-RC-2 and 786-O cells were elevated compared with those from HK-2 cells. In exosomes, GGT1 expression correlated with GGT activity determined using a fluorescent probe for GGT. In RCC patients, serum exosomal GGT activity was elevated in those with advanced stages (III/IV vs. I/II, p = 0.037) and those with microvascular invasion (with vs. without, p = 0.034). Immunohistochemical analysis showed that membranous GGT1 expression was increased in RCC with microvascular invasion. Notably, preoperative serum exosomal GGT activity could predict the likelihood of having microvascular invasion diagnosed by pathological examination of surgically resected specimens. Conclusions: Our results suggest that serum exosomal GGT activity could be a clinically useful marker for advanced clinicopathological features of RCC patients, and its combined use with conventional diagnostic modalities may improve the diagnosis and treatment of patients. [ABSTRACT FROM AUTHOR]

Objectives: Acridone alkaloids from Citrus and their derivatives show various kinds of biological activity. However, the anticancer activities of dimeric acridone alkaloids with unique structures and the molecular mechanism of these effects are poorly understood. Methods: We investigated the cytotoxicity effects of dimeric acridone alkaloids isolated from Marsh grapefruit on human myeloid leukaemia HL‐60 cells. Key findings: Of the six dimeric acridone alkaloids tested, citbismine‐E, the most potent, dose‐ and time‐dependently decreased HL‐60 cell viability by inducing apoptosis. The treatment of HL‐60 cells with citbismine‐E yielded a significant increase in levels of intracellular reactive oxygen species (ROS). Citbismine‐E lowered the mitochondrial membrane potential and increased the activities of caspase‐9 and ‐3. In addition, citbismine‐E‐induced apoptosis, decrease in mitochondrial membrane potential and caspase activation were significantly alleviated by pretreatment of the cells with antioxidant N‐acetylcysteine (NAC). Citbismine‐E induced intrinsic caspase‐dependent apoptosis through ROS‐mediated c‐Jun N‐terminal kinase activation. Citbismine‐E‐induced production of oxidative stress biomarkers, malondialdehyde and 8‐hydroxy‐2′‐deoxyguanosine was also attenuated by pretreatment with NAC. Conclusions: Citbismine‐E is a powerful cytotoxic agent against HL‐60 cells that acts by inducing mitochondrial dysfunction‐mediated apoptosis through ROS‐dependent JNK activation. Citbismine‐E also induced oxidative stress damage via ROS‐mediated lipid peroxidation and DNA damage in HL‐60 cells. [ABSTRACT FROM AUTHOR]

Aims: Sarcopenia is a serious problem because of its poor prognosis. Growth differentiation factor 15 (GDF15) is associated with mitochondrial dysfunction, inflammation, insulin resistance and oxidative stress, which may play crucial roles for the development of sarcopenia. We aimed to examine whether serum GDF15 level is associated with muscle mass, strength and lower extremity function in older patients with cardiometabolic disease. Methods: Serum GDF15 levels were measured in 257 patients with cardiometabolic diseases (including 133 patients with diabetes) who had visited the frailty clinic, using a latex turbidimetric immunoassay. Appendicular skeletal muscle index, handgrip strength, timed‐up‐and‐go test and gait speed were evaluated. Power, speed, balance and total scores based on the sit‐to‐stand test were calculated to assess lower extremity function. Results: The highest tertile of serum GDF15 was independently associated with low handgrip strength, low gait speed, long timed‐up‐and‐go time and scores of lower extremity function but not an appendicular skeletal muscle index in multiple logistic regression analyses after adjustment for covariates. Patients in the highest tertile of GDF15 were at the risk of having three to nine times lower grip strength, three times lower gait speed, five to six times lower mobility and five to 11 times reduction in lower extremity function as compared with those in the lowest GDF15 tertile dependent on the models. Conclusions: Elevated serum GDF15 level was independently associated with low muscle strength and lower extremity function in older patients with cardiometabolic disease. Serum GDF15 could be one of the biomarkers for muscle weakness and low physical performance. Geriatr Gerontol Int 2020; 20: 980–987. [ABSTRACT FROM AUTHOR]

The inactivation mechanism of mould spores using plasma‐generated neutral reactive oxygen species (ROS) was investigated in this paper. Typical cell viability using a counting of colony‐forming unit assay indicated a major state of the spore cells. Both intracellular and extracellular damages have been investigated by a couple of well‐established cell visualization techniques: confocal fluorescence emission microscopy, scanning electron microscopy, and transmission electron microscopy. It is revealed in this paper that the combination of the spore cell viability with those visualization results strongly suggested an early stage of the cell inactivation when ROS can induce intracellular lipid peroxidation through both nanometer‐thick lipid cell membrane and less‐damaged several hundred nanometer‐thick cell wall. [ABSTRACT FROM AUTHOR]

Plasma cell myeloma (PCM) is usually associated with the presence of M‐protein in the serum and urine of patients, and about half of the PCM cases exhibit the IgG M‐protein and increased gamma‐globulin fraction on membrane electrophoresis. The IgG4 subclass is located in the beta‐2 fraction on membrane electrophoresis. The aim of this study was to develop a method to evaluate IgG4‐producing PCM (IgG4‐PCM) and its clinicopathological characteristics. We found three cases of IgG4‐PCM among 80 cases of IgG‐producing PCM by membrane electrophoresis, which were confirmed by IgG4 immunostaining. None of the cases had a clinical history of IgG4‐related disease, although they exhibited high levels of serum IgG4. A bone marrow aspiration specimen had an increased number of plasma cells with a relatively mature morphology. No cases exhibited lymphoplasmacytic inflammation, obliterative phlebitis or fibrosis. Immunohistochemistry revealed that tumor cells expressed CD138 and IgG4 and showed monoclonal expression of kappa. We revealed that IgG4‐PCM might not be associated with IgG4‐related disease and that the detection of M‐protein with beta‐globulin fraction by electrophoresis may be useful for screening IgG4‐PCM. [ABSTRACT FROM AUTHOR]

Background: Vanillin is the main byproduct of alkaline-pretreated lignocellulosic biomass during the process of fermentable-sugar production and a potent inhibitor of ethanol production by yeast. Yeast cells are usually exposed to vanillin during the industrial production of bioethanol from lignocellulosic biomass. Therefore, vanillin toxicity represents a major barrier to reducing the cost of bioethanol production. Results: In this study, we analysed the effects of oxygen-radical treatment on vanillin molecules. Our results showed that vanillin was converted to vanillic acid, protocatechuic aldehyde, protocatechuic acid, methoxyhydroquinone, 3,4-dihydroxy-5-methoxybenzaldehyde, trihydroxy-5-methoxybenzene, and their respective ring-cleaved products, which displayed decreased toxicity relative to vanillin and resulted in reduced vanillin-specific toxicity to yeast during ethanol fermentation. Additionally, after a 16-h incubation, the ethanol concentration in oxygen-radical-treated vanillin solution was 7.0-fold greater than that from non-treated solution, with similar results observed using alkaline-pretreated rice straw slurry with oxygen-radical treatment. Conclusions: This study analysed the effects of oxygen-radical treatment on vanillin molecules in the alkaline-pretreated rice straw slurry, thereby finding that this treatment converted vanillin to its derivatives, resulting in reduced vanillin toxicity to yeast during ethanol fermentation. These findings suggest that a combination of chemical and oxygen-radical treatment improved ethanol production using yeast cells, and that oxygen-radical treatment of plant biomass offers great promise for further improvements in bioethanol-production processes. [ABSTRACT FROM AUTHOR]

The MPN driver mutation, JAK2 exon 12, is rare in MPN and has been exclusively found in polycythaemia vera (PV) (Scott [6]), with no reports of this mutation in MDS/MPN-RS-T to date. We would like to emphasize the importance of analysing JAK2 exon 12 mutation in patients with suspected MDS/MPN-RS-T, but who are negative for JAK2 V617F, CALR exon 9 and MPL W515 mutations. In summary, we identified a patient with MDS/MPN-RS-T harboring SF3B1 and JAK2 exon 12 mutations. Bone marrow morphologies of the patient harboring JAK2 exon 12 mutation, representing features associated with MDS/MPN-RS-T.Figure S3. [Extracted from the article]

Background: Exosomes or small extracellular vesicles secreted from cells are nanovesicles with a diameter of 40–150 nm, which play a number of roles in both physiologic and pathologic processes. In Graves' disease (GD), autoantibodies bind to the thyrotropin receptor (TSHR) on the surface of thyroid follicular epithelial cells and stimulate thyroid growth and thyroid hormone synthesis and secretion via cyclic adenosine monophosphate (cAMP) production. The present study aimed to confirm the existence of TSHR in exosomes secreted from thyroid cells and to define the role of TSHR exosomes in GD. Methods: Exosomes were isolated by differential centrifugation from the culture medium of the human thyroid follicular epithelial cell line (NTHY-ori 3-1) and thyroid carcinoma cell lines (8305C, 8505C, and FTC-133). TSHR expression in cell lysates and exosomes was evaluated by Western blot analysis. In order to study the function of TSHR exosomes, human embryonic kidney (HEK) 293 cells stably expressing TSHR (HEK/TSHR) were established. Using exosomes isolated from both HEK and HEK/TSHR cells, the binding capacity of the M22 human monoclonal autoantibody to TSHR exosomes and their effect on M22-mediated stimulation of cAMP production in HEK/TSHR cells were evaluated. As a positive control for the functional assay, human recombinant TSHR chimera protein capable of binding to TSH was used. Results: TSHR was detected in exosomes from cancer cells as well as normal epithelial cells. An in vitro binding assay showed that alkaline phosphatase-labeled M22 bound to TSHR exosomes in a dose-dependent manner. M22 dose-dependently stimulated intracellular cAMP production in HEK/TSHR cells. The addition of exosomes from HEK/TSHR cells but not those from parental HEK cells significantly ameliorated cAMP production stimulated by treatment with M22 in HEK/TSHR cells. A decoy effect similar to TSHR exosomes was observed for human recombinant TSHR chimera. Conclusions: The results suggest that exosomes expressing TSHR may be secreted from normal and cancerous thyroid cells. In the thyroid gland of patients with GD, TSHR exosomes may exert a decoy effect by sequestering autoantibody, thereby ameliorating autoantibody-mediated activation of thyroid function. [ABSTRACT FROM AUTHOR]

Objective: Prefibrotic/early primary myelofibrosis (pre‐PMF) and essential thrombocythemia (ET) exhibited different features of bone marrow; however, this is not always easy to judge objectively, making pathologists' distinction often suboptimal. In the WHO 2008 criteria, pre‐PMF was not defined as a subgroup of PMF; therefore, affected patients were at a higher risk of misdiagnosis with ET. In this study, we examined the prevalence of pre‐PMF patients among those previously diagnosed with ET in Japan. Method: We reviewed bone marrow specimens and clinical and molecular parameters of patients who were previously diagnosed with ET by the WHO 2008 criteria. Results: Among 107 ET patients, 13 patients were redefined as having pre‐PMF. Pre‐PMF patients exhibited a higher frequency of MPL mutation and increased platelet counts compared to true ET patients. Molecular analysis revealed the frequencies of high‐risk molecular mutations, such as ASXL1, EZH2, and SRSF2, were significantly increased in pre‐PMF patients than those in true ET patients. Conclusion: These results demonstrated the value of reexamining clinical records for patients diagnosed with ET by the WHO 2008 criteria and emphasized that adequate examinations of patients' bone marrow are crucial for an accurate diagnosis of pre‐PMF and ET. [ABSTRACT FROM AUTHOR]

Intravascular large B‐cell lymphoma (IVLBCL) is a distinct disease, but the neoplastic PD‐L1 expression on tumor cells may vary among cases. We evaluated 10 IVLBCL autopsy cases for neoplastic PD‐L1 expression, and had positive results in two cases. In one case, neoplastic PD‐L1 expression (SP142, 28‐8, and E1J2J clones) was dependent on the organ and anatomical site (capillaries vs. vessels) of the tumor tissue. Neoplastic PD‐L1 expression was found in tumor cells located in capillaries in the central nervous system, pituitary gland, kidneys, lung, and gastrointestinal tract; sinuses/sinusoids of the spleen, liver, bone marrow, and lymph nodes; and an extravascular location. However, this expression was not detected in tumor cells located in the adrenal gland, thyroid gland, pancreas, ovaries, uterus, pleura, and small or larger‐sized vessels of the lung. The other case showed constant neoplastic PD‐L1 expression on the tumor cells, and in addition to the affected organs, capillaries, and vessels with two anti‐PD‐L1 antibodies (28‐8 and E1J2J, but not SP142). The divergence and heterogeneity of neoplastic PD‐L1 expression were clearly demonstrated in our cases. To the best of our knowledge, this is the first description of divergent neoplastic PD‐L1 expression among the affected organs and anatomical sites in IVLBCL. [ABSTRACT FROM AUTHOR]

A 64‐year‐old man was admitted with acute onset disturbed consciousness. Cerebrospinal fluid analysis revealed pleocytosis and elevated protein, with negative cultures and PCR. Serum antibodies for autoimmune encephalitis were also negative. Brain magnetic resonance imaging (MRI) was unremarkable, but whole‐body CT scan showed a tumor in the left lower lung lobe. Bronchial brush cytology demonstrated clusters of malignant cells, and 18F‐fluorodeoxyglucose positron emission tomography showed multiple lesions and increased uptake in the lung tumor. Clinically the patient had a stage IV lung carcinoma, graded as T3N3M1b (OSS). Steroid therapy had limited efficacy, but chemotherapy dramatically improved his neurological symptoms. Therefore, he was diagnosed with paraneoplastic autoimmune encephalitis based on the diagnostic criteria for paraneoplastic neurological syndromes. He died due to disease progression 14 months later. Subsequent postmortem examination revealed white ill‐defined nodules in the left lung, with similar nodules in other organs. The brain weighed 1500 g before fixation, and a nodule was observed in the right precentral gyrus. Microscopically, the lung tumor was a pleomorphic carcinoma with an adenocarcinoma component. Multiple areas of micro‐softening (≤500 μm) were identified in the cerebral cortex, gray–white matter junction and basal ganglia, and were distributed diffusely in both the limbic and non‐limbic systems. Mild lymphocytic infiltrates were observed involving few intraparenchymal vessels. Few tumor metastases were observed in the right precentral gyrus. The multiple micro‐softenings may reflect a chronic neuropathologic change of paraneoplastic autoimmune encephalitis. They were too small to be detected by brain MRI. However, these lesions may have the potential to cause the neurological symptoms in the acute phase because they were observed in many anatomical regions. We should pay attention to subtle findings such as micro‐softenings when estimating the neuropathology of autoimmune encephalitis. Further investigations are needed to understand the characteristic neuropathology of this condition. [ABSTRACT FROM AUTHOR]

Aims: Ossification is found occasionally in dedifferentiated liposarcoma (DDLPS). The aims of this study were to elucidate whether the formed bone tissue is usually produced by tumour cells or by reactive non‐neoplastic cells, and to reveal the clinicopathological characteristics of DDLPS with ossification. Methods and results: We examined 36 cases of ossified DDLPS by comparing them to 31 cases of non‐ossified DDLPS. MDM2 amplification was confirmed in osteocytes and/or osteoblastic cells in all but one ossified DDLPS cases (27 of 28) using fluorescence in‐situ hybridisation, although the morphological impression of ossification appeared to be mainly metaplastic (27 of 36) or high‐grade osteosarcoma‐like (six of 36). The bone tissue was often formed predominantly at the periphery of the DDLPS area near the well‐differentiated liposarcoma component (18 of 36), and an organised structure such as bone marrow‐like differentiation was not uncommon (12 of 36). According to a modified French Fédération Nationale des Centers de Lutte Contre le Cancer (FNCLCC) grading system, ossified DDLPS tended to be lower grade than non‐ossified DDLPS (mean grade: 1.88 and 2.15, respectively). Ossification in DDLPS was associated significantly with shorter local recurrence‐free survival by multivariate analysis (P = 0.02347), but metaplastic‐appearing ossification tended to be associated with longer overall survival (P = 0.1400). Conclusions: The bone tissue formed in DDLPS was mainly neoplastic regardless of its morphology and maturity, which highlighted the osteogenic differentiation of the tumour cells. DDLPS patients with osteogenic differentiation tended to suffer from earlier local recurrences, which did not necessarily lead to poor life outcomes. [ABSTRACT FROM AUTHOR]

We isolated a cryptic genospecies of Haemophilus influenzae referred to as ‘ Haemophilus quentini ’ in the urethra of 3 men complaining of urethritis symptoms. H. influenzae strains, which had been isolated from the urethra in 77 of 1518 men complaining of urethritis symptoms, identified by the conventional test, and stored, were re-cultured for this study. Sixty-seven strains surviving storage were screened by a PCR-based assay specific for the cryptic genital Haemophilus genospecies. Three strains (HI09003, HI11006, and HI14016) were screened by PCR and identified as ‘ H. quentini ’ by 16S rRNA sequencing. The men positive for HI09003 and HI11006 were diagnosed as having non-chlamydial non-gonococcal urethritis (NGU), and their demographic and clinical features were similar to those of NGU caused by other pathogens. The man positive for HI14016 was ultimately diagnosed as having condyloma acuminatum on the glans. The 3 strains of ‘ H. quentini ’ produced no β-lactamase and were susceptible to ampicillin and other antimicrobial agents, including cephalosporins, fluoroquinolones, tetracyclines, and macrolides, recommended for treatment for urethritis. ‘ H. quentini ’ would be an uncommon pathogen in men with urogenital infections. Based on the clinical features of the two patients with ‘ H. quentini ’-positive NGU, it would be difficult to predict the presence of ‘ H. quentini ’ in the urethra. The 3 strains of ‘ H. quentini ’ were susceptible to a variety of antimicrobial agents. Further accumulation of data regarding ‘ H. quentini ’ infections is needed to characterize the pathogenic roles of this genospecies in urogenital infections and to establish appropriate management of ‘ H. quentini ’ infections. [ABSTRACT FROM AUTHOR]

Exosomes or microvesicles that are secreted from cells are considered to play important roles in tumor microenvironment. Carbonic anhydrase 9 (CA9), which is induced by hypoxia-inducible factor 1 (HIF1) in response to hypoxia, is overexpressed in many types of cancer including renal cell carcinoma (RCC). We examined the expression level of CA9 in several RCC cell lines and found that the basal level of CA9 was much higher in OSRC-2 cells than in Caki-1, KMRC-1 and 786-O cells. Consistent with the intracellular expression levels, CA9 was abundantly detected in exosomes isolated by ultracentrifugation from OSRC-2 cells. Density gradient centrifugation of OSRC-2 and 786-O exosomes confirmed the co-presence of CA9 with exosomal markers. Upon hypoxia and treatment with CoCl 2 , a hypoxia mimic agent, the CA9 level in exosomes was increased for all cell lines. In order to examine the effects of CA9 exosomes on angiogenesis, we generated stably transfected HEK293 cells expressing CA9. Immunocytochemical staining demonstrated the uptake of CA9 exosomes by human umbilical vein endothelial cells (HUVEC). In vitro angiogenesis assays using HUVEC revealed that CA9 exosomes promoted migration and tube formation. Lastly, MMP2 expression was increased by treatment with CA9 exosomes in HUVEC. Taken together, our results suggest the possibility that CA9 exosomes released from hypoxic RCC may enhance angiogenesis in microenvironment, thereby contributing to cancer progression. [ABSTRACT FROM AUTHOR]

Hypocellular acute myeloid leukemia ( AML) mainly occurs in elderly patients, and is extremely rare in childhood. There is still no established treatment for hypocellular AML. We report the case of an 11-year-old boy with hypocellular AML who was treated successfully with allogenic bone marrow transplantation (allo- BMT). He presented with fever, pallor and pancytopenia. Bone marrow aspiration and biopsy confirmed a diagnosis of hypocellular AML. Although low-dose cytarabine induced reduction of blasts, it did not lead to complete remission. He subsequently received myeloablative conditioning and allo- BMT. Graft-versus-host disease ( GVHD) prophylaxis included short-course methotrexate and cyclosporine. Neutrophil engraftment (>5 × 108/L) and platelet recovery (>10 × 1010/L) were achieved on days 13 and 27, respectively. He developed acute GVHD of the skin (grade 2), which responded well to treatment with prednisolone. He has remained in complete remission for 5 years since allo- BMT. We consider allo- BMT to be feasible for children with hypocellular AML. [ABSTRACT FROM AUTHOR]

We and other have recently shown that growth differentiation factor 15 ( GDF15) is a useful diagnostic marker for mitochondrial diseases, which are inherited disorders caused by mitochondrial or nuclear genomic mutations that lead to impaired energy production. As the primary cause of mitochondrial diseases is mitochondrial dysfunction, the blood level of GDF15 might reflect mitochondrial function in patients, and thus could be a marker for mitochondrial dysfunction. GDF15 has been implicated in aging and various age-related disorders, such as cardiovascular diseases and diabetes, the blood level of which is reportedly elevated in older adults as well as in patients. Although GDF15 might be induced as a result of various cellular stresses and dysfunctions, it would also be possible that the blood GDF15 level reflects at least in part mitochondrial dysfunction in aging and age-related disorders. In the present review, we summarized the current literature regarding GDF15 in aging and age-related disorders from the perspective of biomarkers, with a particular focus on mitochondrial dysfunction. Geriatr Gerontol Int 2016; 16 (Suppl. 1): 17-29. [ABSTRACT FROM AUTHOR]

A wafer temperature feedback control system during plasma processing with rapid, precise, and real-time temperature monitoring employing frequency-domain low-coherence interferometry was developed. To keep the temperature within a specific range, plasma was actively switched on and off, controlled by signals from a monitoring system. It was applied to an organic film etching process with an H2 and N2 mixture gas plasma. The organic material etching yield from atomic hydrogen has a relatively high sensitivity to temperature, and a constant temperature is required to achieve precise etching profiles. This system maintained the wafer temperature within a few degrees for H2/N2 plasma discharges. Duty ratios per discharge gradually decreased because the temperature of the chamber component parts around the wafer increased. The trench width etched in the organic film increased with increasing wafer temperature. This is because of a temperature dependence balance between the etching reaction and protection film formation on the trench sidewall. [ABSTRACT FROM AUTHOR]

Objective: The diagnosis of mitochondrial disorders (MDs) is occasionally difficult because patients often present with solitary, or a combination of, symptoms caused by each organ insufficiency, which may be the result of respiratory chain enzyme deficiency. Growth differentiation factor 15 (GDF-15) has been reported to be elevated in serum of patients with MDs. In this study, we investigated whether GDF-15 is a more useful biomarker for MDs than several conventional biomarkers.Methods: We measured the serum levels of GDF-15 and fibroblast growth factor 21 (FGF-21), as well as other biomarkers, in 48 MD patients and in 146 healthy controls in Japan. GDF-15 and FGF-21 concentrations were measured by enzyme-linked immunosorbant assay and compared with lactate, pyruvate, creatine kinase, and the lactate-to-pyruvate ratio. We calculated sensitivity and specificity and also evaluated the correlation based on two rating scales, including the Newcastle Mitochondrial Disease Rating Scale (NMDAS).Results: Mean GDF-15 concentration was 6-fold higher in MD patients compared to healthy controls (2,711 ± 2,459 pg/ml vs 462.5 ± 141.0 pg/mL; p < 0.001). Using a receiver operating characteristic curve, the area under the curve was significantly higher for GDF-15 than FGF-21 and other conventional biomarkers. Our date suggest that GDF-15 is the most useful biomarker for MDs of the biomarkers examined, and it is associated with MD severity.Interpretation: Our results suggest that measurement of GDF-15 is the most useful first-line test to indicate the patients who have the mitochondrial respiratory chain deficiency. [ABSTRACT FROM AUTHOR]

Primary immunodeficiency disease (PID) is caused by mutations of more than two hundred immunity-related genes. In addition to the heterogeneity of the diseases, the atypical presentation of each disease caused by hypomorphic mutations or somatic mosaicism makes genetic diagnosis challenging. Next-generation sequencing tests all genes simultaneously and has proven its innovative efficacy in genomics. We describe a male PID patient without any family history of immunodeficiency. This patient suffered from recurrent infections from 1 year of age. Laboratory analysis showed hypogammaglobulinemia. T, B, and NK cells were present, but the T cell proliferative response decreased. Whole-exome sequencing analysis identified an IL2RG p.P58T missense mutation. CD8 and CD56 cells showed revertant somatic mosaicism to the wild-type allele. A late-onset and atypical presentation of the X-linked severe combined immunodeficiency (X-SCID) phenotype might be associated with revertant somatic mosaicism in T and NK cells. This patient is the seventh reported case of X-SCID with revertant somatic mosaicism. His classical clinical management did not result in a molecular diagnosis because of the atypical presentation. The coverage that is provided by whole-exome sequencing of most PID genes effectively excluded differential diagnoses other than X-SCID. As next-generation sequencing becomes available in clinical practice, it will enhance our knowledge of PID and rescue currently undiagnosed patients. [ABSTRACT FROM AUTHOR]

The immunological status of patients with Epstein-Barr virus-positive diffuse large B-cell lymphoma (EBV+ DLBCL) without obvious immunodeficiency has not been elucidated. A multicenter prospective study was conducted to assess pretreatment T-cell responses to EBV, EBV-DNA load and anti-EBV antibody in these patients. The proliferative and interferon (IFN)-γ-producing capacity of T-cells in response to autologous B-lymphoblastoid cell lines was determined using carboxyfluorescein diacetate succinimidyl ester (CFSE)-based assay. Frequencies of EBV-specific CD4+T-cells in patients with EBV+DLBCL (n = 13) were significantly higher than in healthy controls (HCs) (n = 16) after both ex vivo and in vitro stimulation. Frequencies of EBV-specific CD8 +T-cells in patients with EBV+ DLBCL tended to be higher than in HCs after in vitro stimulation. Patients with EBV+ DLBCL also showed increased immunoglobulin G (IgG) responses to lytic EBV-encoded antigens. Pretreatment plasma EBV-DNA level was significantly higher in patients with EBV+ DLBCL than in patients with EBV- DLBCL or HCs. In conclusion, EBV-specific T-cells showed increased reactivity, accompanied by higher levels of plasma virus DNA in patients with EBV+ DLBCL. [ABSTRACT FROM AUTHOR]