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Nemaline myopathy (NM), the most common of the congenital myopathies, is caused by various genetic mutations. In this study, we attempted to identify the causative mutations of NM and to reveal any specific genotype-phenotype relationship in Korean patients with this disease. We investigated the clinical features and genotypes in 15 pathologically diagnosed NM patients, using whole exome sequencing (WES) combined with targeted sequencing and array-based comparative genomic hybridization. This strategy revealed pathogenic causative mutations in seven patients (46.7%), among whom mutations in the nebulin gene (NEB) were the most frequent (5 patients, 33.3%). Copy number variation (CNV) abnormality in NEB was not observed in any of our patients. In those with NEB-associated NM, the clinical spectrum was highly variable regardless of the mutation type. However, the majority of patients showing anterior lower leg weakness were associated with mutations located between NEB exons 166 and 177. We concluded that the combination of WES and targeted Sanger sequencing is an effective strategy for analyzing genotypes in patients with NM, and that CNV in NEB may not be a frequent cause of this disease among Koreans.

Episodic ataxia (EA) is a rare neurological condition characterized by recurrent spells of truncal ataxia and incoordination. Five genes (KCNA1, CACNA1A, CACNB4, SLC1A3, and UBR4) have been linked to EA. Despite extensive efforts to genetically diagnose EA, many patients remain still undiagnosed. Whole-exome sequencing was carried out in 39 Korean patients with EA to identify pathogenic mutations of the five known EA genes. We also evaluated 40 candidate genes that cause EA as a secondary phenotype or cerebellar ataxia. Eighteen patients (46%) revealed genetic information useful for establishing a molecular diagnosis of EA. In 11 patients, 16 pathogenic mutations were detected in three EA genes. These included nine mutations in CACNA1A, three in SLC1A3, and four in UBR4. Three patients had mutations in two genes, either CACNA1A and SLC1A3 or CACNA1A and UBR4, suggesting that SLC1A3 and UBR4 may act as genetic modifiers with synergic effects on the abnormal presynaptic activity caused by CACNA1A mutations. In seven patients with negative results for screening of EA genes, potential pathogenic mutations were identified in the candidate genes ATP1A2, SCN1A, TTBK2, TGM6, FGF14, and KCND3. This study demonstrates the genetic heterogeneity of Korean EA, and indicates that whole-exome sequencing may be useful for molecular genetic diagnosis of EA.

The development of ion-assisted aerosol lithography (IAAL) has enabled fabrication of complex three-dimensional nanoparticle (NP) structures on conducting substrates. In this work, the applicability of the IAAL technique was investigated on non-conducting substrates. The NP structure growth process on a non-conducting substrate was found to self-terminate and the structures subsequently repel incoming charged NPs and scatter them away. Electric field calculations supported the experimental findings and confirmed that the electric field distortions owing to charge build-up within the structures prevented additional NP deposition thereon. To regulate the charge build-up without compromising the number of NPs available for assembly, a corona discharger and an ion trap were implemented. By varying the number of ions available in the assembly process, an optimum level of ion injection was found that allowed for a prolonged (>120 min) assembly of NP structures on non-conducting substrates without the unwanted scattering of NPs.

We performed targeted population screening of late onset Pompe disease (LOPD) in unspecified myopathy patients, because early diagnosis is difficult due to its heterogeneous clinical features. We prospectively enrolled 90 unrelated myopathic patients who had one or more signs out of five LOPD-like clinical findings (proximal weakness, axial weakness, lingual weakness, respiratory difficulty, idiopathic hyperCKemia). Acid alpha glucosidase activity was evaluated with dried blood spot and mixed leukocyte simultaneously. For a final diagnosis of LOPD, 16 patients with decreased enzyme activity were genotyped by GAA molecular analysis. We found two patients with LOPD (2.2%), and the remaining 14 patients had at least one G576S or E689K mutation, known as the pseudodeficiency allele. Acid alpha glucosidase activity of LOPD patients was significantly lower than that of patients with at least one pseudodeficiency allele (p = 0.017). This study is the first LOPD screening study for targeted Korean population, and more generally, an Asian population. Our findings suggest that for diagnosis of LOPD in Asian population, modified cutoff value of acid alpha glucosidase activity with dry blood spot considering that of patients having heterozygote pathogenic variants or pseudodeficiency alleles may reduce time and cost requirements and increase the comfort of patients.

Aim Fetal acetylcholine receptor inactivation syndrome (FARIS) occurs in offspring of mothers affected by myasthenia gravis (MG), from in-utero exposure to acetylcholine receptor (AChR)-antibodies targeting the fetal AChR γ-subunit. FARIS causes damage to the fetal neuromuscular junction which is crucial in muscle development, causing a persistent myopathy. FARIS may initially be mistaken for Transient Neonatal Myasthenia Gravis (TNMG), congenital neuromuscular disorders and one of the many causes of neonatal hypotonia. This study aimed to determine the clinical spectrum of FARIS and assess oral salbutamol as a novel pharmacological therapy. Methods Detailed review of antenatal and postnatal clinical features in novel FARIS cases seen in international neuromuscular centres. Antibody data analysis was performed at the Oxford neuroimmunology research laboratory. Oral salbutamol was trialled in five cases based on previously reported benefit in one of our patients. Results We identified 12 novel FARIS cases. At delivery resuscitation was required in all and intubation in nine, all had severe generalised hypotonia. Two infants with arthrogryphosis-multiplex-congenita phenotype died in the neonatal period. Among survivors, there was requirement for mechanical ventilation (n=9), NIPPV (n=2), oxygen (n=1), and supplemental NG/PEG feeding (n=12). The presence of severe generalised hypotonia with dysmorphic features prompted investigations for other neuromuscular, genetic and metabolic disorders which were negative. Common features included facial weakness (n=12) and limb contractures (n=9). Newly described disease features: diaphragmatic paresis (n=5), hearing impairment (n=3), CNS involvement (n=3), pyloric stenosis (n=2), extra-ocular eye restriction (n=2), non-progressive scoliosis (n=2), and jaw opening contracture(n=1). Motor development of patients improved with time. Respiratory complications (tracheostomy;n=2), feeding difficulties (PEG;n=2), facial weakness and speech impairment (from velopharyngeal incompetence) persisted in most. TNMG treatments(immunotherapy/pyridostigmine) were little or no benefit. Novel use of oral salbutamol improved fatiguability, ptosis, oromotor dysfunction, muscle tone, articulation and voice volume in all patients. In 8/12 pregnancies maternal myasthenia gravis hadn’t been established antenatally, and many mothers were pauci/asymptomatic. All had AChR-antibodies targeting the fetal γ-subunit confirming diagnosis. Where subsequent pregnancies were treated aggressively(immunotherapy), infants had improved outcomes. Conclusions This report demonstrates and expands the phenotypic spectrum of FARIS, and emphasises oral salbutamol therapy as a potentially beneficial treatment. FARIS should be considered (mothers or infants tested for fetal specific AChR-Abs) in infants presenting with neonatal hypotonia, myopathic features and/or a suggestive antenatal history, even in the absence of a maternal MG diagnosis. Aggressive treatment with immunotherapy in pregnancy may improve outcomes.

The results of clinical trials using mesenchymal stem cells (MSCs) are controversial due to the heterogeneity of human MSCs and differences in culture conditions. In this regard, it is important to identify gene expression patterns according to culture conditions, and to determine how the cells are expanded and when they should be clinically used. In the current study, stemness gene expression was investigated in adipose tissue-derived MSCs (AT-MSCs) harvested following culture at different densities. AT-MSCs were plated at a density of 200 or 5,000 cells/cm2. After 7 days of culture, stemness gene expression was examined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis. The proliferation rate of AT-MSCs harvested at a low density (~50% confluent) was higher than that of AT-MSCs harvested at a high density (~90% confluent). Although there were differences in the expression levels of stemness gene, such as octamer-binding transcription factor 4, nanog homeobox (Nanog), SRY-box 2, Kruppel like factor 4, v-myc avian myelocytomatosis viral oncogene homolog (c-Myc), and lin-28 homolog A, in the AT-MSCs obtained from different donors, RT-qPCR analysis demonstrated differential gene expression patterns according to the cell culture density. Expression levels of stemness genes, particularly Nanog and c-Myc, were upregulated in AT-MSCs harvested at a low density (~50% confluent) in comparison to AT-MSCs from the same donor harvested at a high density (~90% confluent). These results imply that culture conditions, such as the cell density at harvesting, modulate the stemness gene expression and proliferation of MSCs.

Aerosol particles generated by spark discharge methods generally have a Gaussian charge distribution centered at zero, and these charges come from the plasma between the electrodes as well as the ions generated during the process. However, some particle manipulation applications demand control in charges of the particles, for instance, in electrostatic deposition of metal nanoparticles through ion assisted aerosol lithography (IAAL). Neutralizing and re-charging the particles to a particular charge leads to low yield, especially at particle sizes below 10 nm. Therefore, we have devised a new strategy to increase the yield of positively charged sub-10 nm particles, by continuously providing additional positive ions to charge the generated particles in situ, using the electrodes as the ion source. To this end, we have designed a new spark control circuit that rapidly restores corona discharges right after a spark discharge event. In the new circuit, an additional voltage source connected in series with the anode restores its electric potential to the corona discharge voltage. When used with pin-to-plate electrode configuration, the amount of positively charged particles increased by up to 1.8-fold while negatively charged particles were reduced by half, at an optimum voltage set point of 3400 V. Rod-to-plate and wire-in-hole configurations were also tested with the new circuit to investigate the effect of electrode geometry. Lastly, the effect of carrier gas flow rate was studied to confirm that the particles were indeed charged by positive ions from corona discharges.

Scaling up the production capacity of spark discharge generated nanoparticles is crucial for their industrial application. Among various options, high-frequency spark discharge generation is a promising route as the mass production rate of nanoparticles should simply scale linearly with the spark frequency. However, reports of spark discharge generators operating above 1 kHz are scarce in the literature, as spark discharges at higher frequencies have been observed to be irregular, leading to premature spark discharges that result in a significantly lower mass production rate. In this study, we present a wire-to-plate electrode configuration that suppresses premature spark discharges during high-frequency operations above 1 kHz, and investigate the factors that contribute to the occurrence of premature spark discharges by comparing the performance of the wire-to-plate electrode configuration to those of rod-to-rod and wire-to-rod configurations. We identified that spark duration should be minimized...

This study aimed to develop a new set of screening criteria that is easily applicable and highly sensitive for the detection of patients at high risk of Fabry disease (FD) among hypertrophic cardiomyopathy (HCM) patients. We prospectively studied 273 consecutive unrelated patients who were referred to HCM clinic for unknown left ventricular hypertrophy. Among the 273 patients, we selected 65 high-risk patients who fulfilled at least one of our newly proposed screening criteria. All 273 patients were assayed for plasma α-galactosidase A (α-GAL A) activity. The new screening criteria were: (1) atypical HCM, (2) history or presence of documented arrhythmia, (3) short PR interval defined as

Cr–Ni–N coatings, the Ni content of which was altered from 0 to 6·3 at-%, were deposited by a hybrid coating system consisting of high power impulse magnetron sputtering and radio frequency magnetron sputtering. The effects of Ni addition to Cr-N coatings on the microstructure and mechanical properties of the coatings were investigated in this study. The instrumental analysis revealed that the Ni element was incorporated into Cr-N crystals as solid solutions, while excess Ni was precipitated as nanocrystalline phases at the Cr-N grain boundaries. The toughness of the Cr–Ni–N coatings was significantly improved with Ni content increased. In addition, the hardness of the coatings slightly increased from 30·4 to 32·6 GPa due to the grain refinement effect and solid solution hardening. From the tribological tests, the highest wear resistance was obtained from the sample having a Ni content of 2·7 at-%.

FHL1-related myopathies are clinically heterogeneous, involving skeletal and cardiac muscles. Overlapping clinical features include joint contractures, rigid spine, scapuloperoneal weakness and cardiac diseases. Histopathologically, reducing bodies are the most characteristic finding, but not present in all FHL1-related cases. Non-specific dystrophic pathology without reducing body is usual in the forms of X-linked myopathy with postural muscle atrophy, Emery-Dreifuss muscular dystrophy and isolated hypertrophic cardiomyopathy. Here, we describe a patient with mild weakness with ankle contracture. We finally concluded he has a FHL1-related myopathy at an extreme end of phenotypic spectrum of FHL1 myopathy, which one might miss to recognize as a form of myopathy. The genetic variant was detected by whole exome sequencing, and its pathogenicity was clearly confirmed with pathological and biochemical studies. This is the first FHL1 case with a mildest phenotype backed by biochemical/genetic evidence. This report will help clinicians hesitating to further evaluate mild cases to better correlate the genotype to the phenotype.

Objectives This study aims to explore the roles of N‐myc and caspase‐8 in TRAIL‐resistant IMR‐32 cells which exhibit MYCN oncogene amplification and lack caspase‐8 expression. Materials and methods We established N‐myc–downregulated IMR‐32 cells using shRNA lentiviral particles targeting N‐myc and examined the effect the N‐myc inhibition on TRAIL susceptibility in human neuroblastoma IMR‐32 cells expressing caspase‐8. Results Cisplatin treatment in IMR‐32 cells increased the expression of death receptor 5 (DR5; TRAIL‐R2), but not other receptors, via downregulation of NF‐κB activity. However, the cisplatin‐mediated increase in DR5 failed to induce cell death following TRAIL treatment. Furthermore, interferon (IFN)‐γ pretreatment increased caspase‐8 expression in IMR‐32 cells, but cisplatin failed to trigger TRAIL cytotoxicity. We downregulated N‐myc expression in IMR‐32 cells using N‐myc–targeting shRNA. These cells showed decreased growth rate and Bcl‐2 expression accompanied by a mild collapse in the mitochondrial membrane potential as compared with those treated with scrambled shRNA. TRAIL treatment in N‐myc–negative cells expressing caspase‐8 following IFN‐γ treatment significantly triggered apoptotic cell death. Concurrent treatment with cisplatin enhanced TRAIL‐mediated cytotoxicity, which was abrogated by an additional pretreatment with DR5:Fc chimera protein. Conclusions N‐myc and caspase‐8 expressions are involved in TRAIL susceptibility in IMR‐32 cells, and the combination of treatment with cisplatin and TRAIL may serve as a promising strategy for the development of therapeutics against neuroblastoma that is controlled by N‐myc and caspase‐8 expression.

In this study, we investigated the effect of cell density on the proliferation activity of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT-MSCs) over time in culture. Passage #4 (P4) and #12 (P12) AT-MSCs from two donors were plated at a density of 200 (culture condition 1, CC1) or 5000 (culture condition 2, CC2) cells cm(-2) . After 7 days of incubation, P4 and P12 AT-MSCs cultured in CC1 were thin and spindle-shaped, whereas those cultured in CC2 had extensive cell-to-cell contacts and an expanded cell volume. In addition, P4 and P12 AT-MSCs in CC1 divided more than three times, while those in CC2 divided less than once on average. Flow cytometric analysis using 5(6)-carboxyfluorescein diacetate N-succinimidyl ester dye showed that the fluorescence intensity of AT-MSCs was lower in CC1 than in CC2. Furthermore, expression of proliferation-associated genes, such as CDC45L, CDC20A and KIF20A, in P4 AT-MSCs was higher in CC1 than in CC2, and this difference was also observed in P12 AT-MSCs. These data demonstrated that cell culture density affects the proliferation activity of MSCs, suggesting that it is feasible to design a strategy to prepare suitable MSCs using specific culture conditions.

Human mesenchymal stem cell (MSC) heterogeneity and problems associated with the ex vivo expansion of MSC are linked with the failure of MSC clinical trials. In this study, we compared the effect of MSCs cultured in different oxygen concentrations on GVHD in mice to elucidate whether hypoxia improves the immunosuppressive capacity of MSCs. Hypoxia increased the proliferative activity and the expression of several stemness and chemokine genes, such as KLF4, OCT4, C-MYC, CCL2, and CXCL10. Mice that received MSCs cultured in normoxia or hypoxia showed alleviated symptoms of GVHD and increased survival times. However, there was no significant difference in survival rates between mice that received MSCs cultured in normoxia and hypoxia. These data suggest that hypoxic culture is a useful method for maintaining and obtaining MSCs used for cell therapy and that the therapeutic potential of MSCs cultured in hypoxia warrants further investigation.

We demonstrate controlled printing of charged nanoparticles using a metal coated stencil mask by applying an electric potential to engineer the electric field streamlines through the mask opening. The potential difference between the metal coated mask and the substrate generates an electrostatic lens effect which focuses the charged nanoparticles toward the center of the opening and hence reduces the size of the printed nanoparticle clusters. In contrast to previously reported ion induced focusing approach, the present method does not rely on ion accumulation, but simply requires changing the potential difference between the mask and the substrate to control the focusing of charged aerosols. By adjusting the potential difference between the mask and the substrate, electric field distortion near the mask opening can be precisely controlled. Using this approach, the printed patterns can be scaled down by up to a factor of 7.3 in each dimension from the mask opening, which enables printing of sub-micrometer sized particle clusters using a mask with micrometer scale opening sizes. Particle trajectories were calculated by solving the Langevin equation, and the resulting particle deposition profile was compared with the experimental results. Using this approach, a multi-material nanoparticle cluster array was fabricated by a sequential deposition of silver and copper nanoparticles after lateral translation of the mask, resulting in offset arrays of silver and copper nanoparticle clusters on the same substrate.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), either alone or in combination with other anti-cancer agents, is a promising new strategy for the treatment of cancer. However, aberrant PI3K/Akt/mTOR survival signaling may confer TRAIL resistance by altering the balance between pro- and anti-apoptotic proteins. In the present study, we showed that the Akt/mTOR inhibitor RAD001 (everolimus) induced cell death in a dose-dependent manner and enhanced TRAIL-induced apoptosis in human leukemic Jurkat T cells, which show PI3K/Akt/mTOR pathway activation and basal expression levels of death receptor (DR) 5 (TRAIL-R2). Investigation of the effect of RAD001 treatment on the expression of TRAIL receptors (TRAIL-Rs) in Jurkat T cells showed that RAD001 significantly upregulated DR5 by up to 51.22%, but not other TRAIL-Rs such as DR4 (TRAIL-R1), decoy receptor (DcR) 1 (TRAIL-R3), and DcR2 (TRAIL-R4). Pretreatment with DR5:Fc chimera abrogated the RAD001-induced increase of TRAIL cytotoxicity, indicating that the upregulation of DR5 by RAD001 plays a role in enhancing the susceptibility of Jurkat T cells to TRAIL. Our results indicate that combination treatment with RAD001 and TRAIL may be a novel therapeutic strategy in leukemia.

In our previous studies, the recombinant type II macrophage migration inhibitory factor homologue (rAs-MIF) secreted from Anisakis simplex suppressed experimental inflammation mouse model through IL-10 production and CD4(+)CD25(+)Foxp3(+) T-cell recruitment. Also, TLR2 gene expression was significantly increased following rAs-MIF treatment. To know the relation between TLR2 and amelioration mechanisms of rAs-MIF, we induced allergic airway inflammation by ovalbumin and alum with or without rAs-MIF under TLR2 blocking systems [anti-TLR2-specific antibody (α-mTLR2 Ab) treatment and using TLR2 knockout mice]. As a result, the amelioration effects of rAs-MIF in allergic airway inflammation model (diminished inflammation and Th2 response in the lung, increased IL-10 secretion, CD4(+)CD25(+)Foxp3(+) T-cell recruitment) were diminished under two of the TLR2 blocking model. The expression of TLR2 on the surface of lung epithelial cell was significantly elevated by rAs-MIF treatment or Pam3CSK (TLR2-specific agonist) treatment, but they might have some competition effect on the elevation of TLR2 expression. In addition, the elevation of IL-10 gene expression by rAs-MIF treatment was significantly inhibited by α-mTLR2 Ab or Pam3CSK pretreatment. In conclusion, anti-inflammatory effects of the rAs-MIF on OVA-induced allergic airway inflammation might be closely related to TLR2.

The use of soft X-rays in a neutralizer represents an alternative technique that could replace conventional radioactive sources. In this study, we evaluated the charging characteristics of a soft X-ray aerosol neutralizer. In addition, the results from the evaluation of the soft X-ray charger were compared with results obtained using a neutralizer incorporating an 241Am radioactive source. The tandem differential mobility analyzer technique was used previously to determine the size-dependent positive, negative, and neutral charge fractions of a soft X-ray neutralizer. This technique was used to show that the neutral fractions obtained using the soft X-ray charger agreed well with the predictions of bipolar diffusion charging theory, and that the soft X-ray charger could be used as a neutralizer for a scanning mobility particle sizer system.

// Myoung Woo Lee 1, 2, * , Somi Ryu 3, * , Dae Seong Kim 1, 2 , Ji Won Lee 1 , Ki Woong Sung 1 , Hong Hoe Koo 1, 2, 4 and Keon Hee Yoo 1, 2, 5 1 Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea 2 Stem Cell and Regenerative Medicine Institute, Samsung Medical Center, Seoul, Korea 3 Gyeongsang National University Hospital, Gyeongsang National University School of Medicine, Jinju, Korea 4 Department of Health Sciences and Technology, SAIHST, Sungkyunkwan University, Seoul, Korea 5 Department of Medical Device Management and Research, SAIHST, Sungkyunkwan University, Seoul, Korea * These authors contributed equally to this work Correspondence to: Keon Hee Yoo, email: hema2170@skku.edu Keywords: mesenchymal stem/stromal cell; hematologic malignancy; leukemia; anti-cancer therapy; cell therapy Received: August 29, 2017 Accepted: November 16, 2017 Published: January 02, 2018 ABSTRACT Mesenchymal stem/stromal cells (MSCs) are known for being multi-potent; however, they also possess anti-cancer properties, prompting efforts to adapt MSCs for therapies. However, MSCs have also been widely implicated in pathways that, on the contrary, contribute to tumor growth. Numerous studies have been conducted to adapt MSCs for further clinical use, but the results have been inconclusive, possibly due to the heterogeneity of MSC populations. Moreover, the conflicting roles of MSCs in tumor inhibition and tumor growth impede their adaptation for anti-cancer therapies. Anti-tumorigenic and pro-tumorigenic properties of MSCs in hematologic malignancies are not as well established as they are for solid malignancies and data comparing them are still limited. In this review, cancer-related influences of MSCs on hematologic malignancies, such as leukemia and lymphoma, are discussed in addition to underlying mechanisms, as well as sources of MSCs and effects on different cancer types. This review describes how MSCs preserve both anti- and pro-tumorigenic effects, as they tend to not only inhibit tumor growth by suppressing tumor cell proliferation, but also promote tumor growth by suppressing tumor cell apoptosis. Thus, clinical studies trying to adapt MSCs for anti-cancer therapies should consider that MSCs could actually promote hematologic cancer progression. It is necessary to take extreme care in developing MSC-based cell therapies in order to boost anti-cancer properties while eliminating tumor-favoring effects. This review emphasizes that research regarding therapies with MSCs must consider the fact that they exert both anti- and pro-tumorigenic effects on hematologic malignancies.

Human mesenchymal stem cells (MSCs) are known for their prostaglandin E2 (PGE2)-mediated immunosuppressive function but the precise molecular mechanisms underlying PGE2 biosynthesis during inflammation have not been completely elucidated. In this study, we have investigated the involvement of PGE2 pathway members in PGE2 production by bone marrow (BM)-MSCs in response to inflammatory stimuli, and their role in immunosuppression mediated by BM-MSCs. We found that IFN-γ and TNF-α increased cyclooxygenase (COX)-2 expression but not that of prostaglandin E synthase (PGES), or PGE2 production. On the other hand, the toll like receptor 3 (TLR3) stimulant poly(I:C) increased expression of both COX-2 and PGES, resulting in a significant increase in PGE2 levels. This effect was reversed upon COX-2 inhibition with indomethacin or PGES downregulation by siRNA. Reduced PGE2 levels decreased MSC's capacity to inhibit hPBMC proliferation. In addition, administration of MSCs with inhibited PGES expression into mice with graft-versus-host disease (GVHD) did not reduce mortality. In summary, the present study reveals that upregulation of PGES via TLR3 is critical for BM-MSCs-mediated immunosuppression by PGE2 secretion via the COX-2/PGE2 pathway. These results provide a basis for understanding the molecular mechanisms underlying the PGE2-mediated immunosuppressive properties of MSCs.

Mesenchymal stem cells (MSCs) are of particular interest for the treatment of immune-related diseases owing to their immunosuppressive properties. In this study, we aimed to identify the effect of interferon (IFN)-γ priming on immunomodulation by MSCs and elucidate the possible mechanism underlying their properties for the clinical treatment of allogeneic conflicts. Infusion of MSCs primed with IFN-γ significantly reduced the symptoms of graft-versus-host disease (GVHD) in NOD-SCID mice, thereby increasing survival rate when compared with naïve MSC-infused mice. However, infusion of IFN-γ-primed MSCs in which indoleamine 2,3-dioxygenase (IDO) was downregulated did not elicit this effect. The IDO gene was expressed in MSCs via the IFN-γ-Janus kinase (JAK)-signal transducer and activator of transcription 1 (STAT1) pathway, and the infusion of IDO-over-expressing MSCs increased survival rate in an in vivo GVHD model, similar to infusion of IFN-γ-primed MSCs. These data indicate that IFN-γ production by activated T-cells is correlated with the induction of IDO expression in MSCs via the IFN-γ-JAK-STAT1 pathway, which in turn results in the suppression of T-cell proliferation. Our findings also suggest that cell therapy based on MSCs primed with IFN-γ can be used for the clinical treatment of allogeneic conflicts, including GVHD., Highlights • IFN-γ priming enhances the immunosuppressive properties of human MSCs in in vitro and in vivo models. • IFN-γ priming induces IDO expression in MSCs via the JAK/STAT1 signaling pathway, but TLR3 activation does not. • Cell therapy using MSCs primed with IFN-γ could be highly effective in treating allogeneic diseases, including GVHD. It is necessary to improve the function of mesenchymal stem cells (MSCs) to maximize their treatment potential beyond what is currently achieved in cell therapy studies using naïve heterogeneous MSCs. The preclinical study of a candidate cell therapy based on MSCs primed with interferon-γ as reported in this study, could lay the foundation for the use of cell therapy for the treatment of graft-versus-host disease (GVHD), and is very important for the initiation of clinical trials. Our findings also suggest that cell therapy based on functionally improved MSCs could be used for the clinical treatment of allogeneic conflicts.

The peroxisome proliferator-activated receptor (PPAR) γ, a subtype of PPARs, is a member of the nuclear receptor family. PPARγ and its ligands contribute to various types of diseases including cancer. Given that currently developed therapies against leukemia are not very effective or safe, PPARγ ligands have been shown to be a new class of compounds with the potential to treat hematologic malignancies, particularly leukemia. The capability of PPARγ ligands to induce apoptosis, inhibit proliferation, and promote differentiation of leukemia cells suggests it has significant potential as a drug against leukemia. However, the specific mechanisms and molecules involved are not well-understood, although a number of PPARγ ligands with anti-leukemic effects have been identified. This may explain why PPARγ ligands have not been widely evaluated in clinical trials. To fill the gaps in the lack of understanding of specific anti-leukemic processes of PPARγ ligands and further adapt these molecules as anti-leukemic agents, this review describes previous studies of the anti-leukemic effects of PPARγ ligands.

Supplemental Digital Content is available in the text, Fabry disease is a rare X-linked lysosomal storage disorder caused by an α-galactosidase A deficiency. The progressive accumulation of globotriaosylceramide (GL-3) results in life-threatening complications, including renal, cardiac, and cerebrovascular diseases. This study investigated the phenotypic and molecular spectra of GLA mutations in Korean patients with Fabry disease using a nationwide survey. This study included 94 patients from 46 independent pedigrees: 38 adult males, 46 symptomatic females, and 10 pediatric males. Each diagnosis was based on an enzyme assay and GLA gene mutation analysis. The mean age at presentation was 24 years (range, 5–65 years); however, the diagnoses were delayed by 21 ± 19 years after the onset of symptoms. Those patients with late-onset Fabry disease were diagnosed by family screening or milder symptoms at a later age. Forty different mutations were identified: 20 missense (50%), 10 nonsense (25%), 8 frameshift (20%), and 2 splice site (5%) mutations. Five of them were novel. IVS4+919G>A (c.936+919 G>A) was not detected among the 6505 alleles via newborn screening using dried blood spots. Enzyme replacement therapy (ERT) was performed in all the males and pediatric patients, whereas 75% of the symptomatic females underwent ERT for 4.2 ± 3.6 years. This study described the demographic data, wide clinical spectrum of phenotypes, and GLA mutation spectrum of Fabry disease in Korea. Most of the patients had classical Fabry disease, with a 4 times higher incidence than that of late-onset Fabry disease, indicating an underdiagnosis of mild, late-onset Fabry disease.

Late-onset Pompe disease (LOPD) is an autosomal recessive disorder caused by deficiency of the enzyme acid glucosidase alfa (GAA). Recently, enzyme replacement therapy (ERT) using recombinant human GAA (rhGAA) became clinically available, and is expected to modify the clinical course in LOPD of various stages. In this study, we evaluated the efficacy and adverse events of ERT for 48 weeks in Korean LOPD patients. Five Korean LOPD patients were included in the study. At baseline, clinical and laboratory features, including muscular and pulmonary function, were assessed, and rhGAA was infused every 2 weeks. Then, patients were examined at every 12-week interval for evaluation of changes in motor and pulmonary function for 48 weeks along with adverse reactions to ERT. The muscular and pulmonary function of the patients varied depending on the baseline condition of the patient after 48 weeks of ERT. However, the overall function of the patients showed stabilization of the disease rather than the improvement seen in infantile-onset Pompe disease. This is the first clinical study on ERT of Korean LOPD patients. Results of our study showed stabilization of muscular and pulmonary function in LOPD patients for 48 weeks with a favorable prognosis for patients who received early diagnosis and ambulatory patients. One of our patients developed a serious anaphylactic reaction, which necessitated the cessation of further ERT. Therefore, our study shows that early diagnosis and ERT are important in preventing further deterioration of the disease.

Background Central core disease (CCD) is a congenital myopathy characterized by distinctive cores in muscle fibers. Mutations in the gene encoding ryanodine receptor 1 (RYR1) have been identified in most CCD patients. Case report Two unrelated patients presented with slowly progressive or nonprogressive proximal muscle weakness since childhood. Their family history revealed some members with the same clinical problem. Histological analysis of muscle biopsy samples revealed numerous peripheral cores in the muscle fibers. RYR1 sequence analysis disclosed a novel mutation in exon 101 (c.14590T>C) and confirmed a previously reported mutation in exon 102 (c.14678G>A). Conclusions We report herein two families with CCD in whom missense mutations at the C-terminal of RYR1 were identified. Although it has been accepted that such mutations are usually associated with a severe clinical phenotype and clearly demarcated central cores, our patients exhibited a mild clinical phenotype without facial muscle involvement and skeletal deformities, and atypical cores in their muscle biopsy specimens.

PURPOSE The underlying cause of myasthenia gravis (MG) is unknown, although it likely involves a genetic component. However, no common genetic variants have been unequivocally linked to autoimmune MG. We sought to identify the genetic variants associated with an increased or decreased risk of developing MG in samples from a Korean Multicenter MG Cohort. MATERIALS AND METHODS To determine new genetic targets related to autoimmune MG, a whole genome-based single nucleotide polymorphisms (SNP) analysis was conducted using an Axiom™ Genome-Wide ASI 1 Array, comprising 598375 SNPs and samples from 109 MG patients and 150 neurologically normal controls. RESULTS In total, 641 SNPs from five case-control associations showed p-values of less than 10⁻⁵. From regional analysis, we selected seven candidate genes (RYR3, CACNA1S, SLAMF1, SOX5, FHOD3, GABRB1, and SACS) for further analysis. CONCLUSION The present study suggests that a few genetic polymorphisms, such as in RYR3, CACNA1S, and SLAMF1, might be related to autoimmune MG. Our findings also encourage further studies, particularly confirmatory studies with larger samples, to validate and analyze the association between these SNPs and autoimmune MG.

Background and Objectives Transplantation of cryopreserved umbilical cord blood (UCB) can be used to treat a multitude of haematologic and immunological diseases. In this study, we examined the quality of UCB cryopreserved for 2 (group I), 4 (group II) and 6 (group III) years. Methods The following parameters and procedures were used to test individual units of cryopreserved UCB: the number of total nucleated cells (TNC), cell viability, CFU-GM assay, T-cell activation in vitro and haematopoietic stem cell engraftment in NOD/SCID mice in vivo. Results The TNC recovery rates for groups I, II and III were 106·2 ± 6·17%, 96·69 ± 6·39% and 100·38 ± 5·27%, respectively, and the mean percentages of viable cells after thawing were 86·88%, 86·38% and 87·43%. When TNC were plated at 5 × 103, the number of CFU-GM was 13·6 (group I), 13·8 (group II), 14·2 (group III) and 14·7 (fresh UCB). We confirmed that the huCD4+ and huCD8+ T cells within cryopreserved UCB are functionally responsive by assessment of activated huCD25+ cells. Moreover, the percentage of huCD45+ cells in the bone marrow was 4·32 ± 1·29% (group I), 4·48 ± 1·11% (group II), 4·40% ± 1·12% (group III) and 4·50% ± 0·66% (fresh UCB), and that in the peripheral blood was 14·69 ± 3·08% (group I), 15·24 ± 4·05% (group II), 15·74 ± 3·43% (group III) and 17·48 ± 3·74% (fresh UCB) in NOD/SCID mice infused with isolated huCD34+ cells. Conclusion These results indicated that cryopreserved UCB units efficiently retrieve in functionally competent form and are suitable for transplantation.

Pearson syndrome. Clinical spectrum of mitochondrial disorders is broad, so consensus diagnostic criteria for mitochondrial disorders have been proposed for children and adults. Sequence map of human mitochondrial genome with its normal and patho­ genic variants is publicly available. In this review, we will discuss genetic features of mitochondrial myopathies as well as their key findings. Representative syndromes will be described in detail and the less common entities will be summarized in Table 1.

Anticipation is a phenomenon characterized by decreasing age at onset and increasing severity of symptoms of a disease in successive generations within a pedigree. Anticipation mostly occurs in neurodegenerative diseases with expansion of unstable trinucleotide repeats. However, it has not been previously pointed out in episodic ataxia type 2 (EA2). Clinical and genetic analyses were performed in nine members from three consecutive generations of a Korean family with EA2. We performed a polymerase chain reaction (PCR)-based direct sequence analysis of all coding regions of CACNA1A using genomic DNA. The clinically affected family members showed recurrent vertigo, interictal nystagmus, and childhood epilepsy. There is a decrease in the age onset (possible genetic anticipation) in three succeeding generations of the family. Genetic analysis identified a splice site mutation (p.Val1465Glyfs13X) and normal trinucleotide repeats in CACNA1A in all clinically affected and one unaffected members. Recognizing anticipation would aid in genetic counseling in EA2.