Development of cost-effective and fast KIR genotyping by multiplex PCR-SSP.

Killer-cell immunoglobulin-like receptors (KIR) control natural killer (NK) cell functions by recognizing HLA molecules and modulating the activity of NK cells. The KIR gene cluster contains polymorphic and highly homologous genes. Diversity of the KIR region is achieved through differences in gene content, allelic polymorphism, and gene copy number, which result in unrelated individuals having different KIR genotypes and individualized immune responses that are relevant to multiple aspects of human health and disease. Therefore, KIR genotyping is increasingly used in epidemiological studies. Here, we developed multiplex polymerase chain reaction with sequence-specific primers (PCR-SSP) to compensate for the shortcomings of the conventional PCR-SSP method, which is most commonly used for KIR analysis. Multiplex PCR-SSP method involves six multiplex reactions that detect 16 KIR genes and distinguish variant types of some KIR genes by adding two reactions. The assay was evaluated in a blind survey using a panel of 40 reference DNA standards from the UCLA KIR Exchange Program. The results are 100% concordant with the genotype determined using Luminex-based reverse sequence-specific oligonucleotide typing systems. Additionally, we investigated the currently known 16 KIR genes and their common variants in 120 unrelated Korean individuals. The results were consistent with the KIR genotype previously reported by Hwang et al. This multiplex PCR-SSP is an efficient method for analyzing KIR genotypes in both small- and large-scale studies with minimal labor, reagents, and DNA. Furthermore, by providing a better definition of KIR polymorphisms it can contribute to developments in immunogenetics.

Assessment of a new genomic classification system in acute myeloid leukemia with a normal karyotype.

This study was performed to assess if a recently recommended genomic classification is predictive in patients with normal-karyotype (NK) acute myeloid leukemia (AML). A total of 393 patients were included. Analysis of genetic mutations was performed using targeted resequencing with an Illumina Hiseq 2000. We identified driver mutations across 40 genes, with one or more driver mutations identified in 95.7% of patients. The molecular subclassification was as follows: 34.6% patients (n = 136) with AML with the NPM1 mutation, 10.7% (n = 42) with AML with mutated chromatin or RNA-splicing genes or both, 1.5% (n = 6) with AML with TP53 mutations, 13.5% (n = 53) with AML with biallelic CEBPA mutations, 2.0% (n = 8) with AML with IDH2-R172 mutations and no other class-defining lesion, 29.5% (n = 116) with AML with driver mutations but no detected class-defining lesion, 4.3% (n = 17) with AML with no detected driver mutation, and 3.8% (n = 15) patients with AML who met the criteria for ≥2 genomic subgroups. The 5-year overall survival and relapse rate of subgroup in AML with mutated chromatin, RNA-splicing genes, or both was 11.6% (95% CI = 1.4-21.8%) and 71.4% (95% CI = 45.7-86.5%), respectively. This study suggests that the recently recommended genomic classification is an appropriate and replicable categorization system in the NK AML population. The subgroup of AML with mutated chromatin, RNA-splicing genes, or both showed extremely poor survival in NK-AML; thus, a novel approach is needed to improve their prognosis.

Enhancing the photovoltaic performance and stability of QDSSCs using surface reinforced Pt nanostructures with controllable morphology and superior electrocatalysis via cost-effective chemical bath deposition.

To make quantum-dot sensitized solar cells (QDSSCs) competitive, photovoltaic parameters such as the power conversion efficiency (PCE) and fill factor (FF) must become comparable to those of other emerging solar cell technologies. In the present study, a novel strategy has been successfully developed for a highly efficient surface-modified platinum (Pt) counter electrode (CE) with high catalytic activity and long-term stability in a polysulfide redox electrolyte. The reinforcement of the Pt surface was performed using a thin passivating layer of CuS, NiS, or CoS by simple chemical bath deposition techniques. This method was a more efficient method for reducing the electron recombination in QDSSCs. The optimized Pt/CuS CE shows a very low charge transfer resistance of 37.01 Ω, which is an order of magnitude lower than those of bare Pt (86.32 Ω), Pt/NiS (53.83 Ω), and Pt/CoS (73.51 Ω) CEs. Therefore, the Pt/CuS CEs show much greater catalytic activity in the polysulfide redox electrolyte than Pt, Pt/NiS and Pt/CoS CEs. As a result, under one-sun illumination (AM 1.5G, 100 mW cm(-2)), the Pt/CuS CE exhibits a PCE of 4.32%, which is higher than the values of 1.77%, 2.95%, and 3.25% obtained with bare Pt, Pt/CoS, and Pt/NiS CEs, respectively. The performance of the Pt/CuS CE was enhanced by the improved current density, Cu vacancies with increased S composition, and surface morphology, which enable rapid electron transport and lower the electron recombination rate for the polysulfide electrolyte redox couple. Electrochemical impedance spectroscopy and Tafel polarization revealed that the hybrid CEs reduce interfacial recombination and exhibit better electrochemical and photovoltaic performance compared with a bare Pt CE. The Pt/CuS CE also shows superior stability in the polysulfide electrolyte in a working state for over 10 h, resulting in a long-term electrode stability than Pt CE.

Enhanced photovoltaic performance and time varied controllable growth of a CuS nanoplatelet structured thin film and its application as an efficient counter electrode for quantum dot-sensitized solar cells via a cost-effective chemical bath deposition.

For the first time we report a simple synthetic strategy to prepare copper sulfide counter electrodes on fluorine-doped tin oxide (FTO) substrates using the inexpensive chemical bath deposition method in the presence of hydrochloric acid (HCl) at different deposition times. CuS nanoplatelet structures were uniformly grown on the FTO substrate with a good dispersion and optimized conditions. The growth process of the CuS nanoplatelets can be controlled by changing the deposition time in the presence of HCl. HCl acts as a complexing agent as well as improving S(2-) concentration against S atoms in this one-step preparation. The photovoltaic performance was significantly improved in terms of the power conversion efficiency (PCE), short-circuit density (J(sc)), open-circuit voltage (V(oc)), and the fill factor (FF). The optimized deposition time of CuS 60 min resulted in a higher PCE of 4.06%, J(sc) of 12.92 mA cm(-2), V(oc) of 0.60 V, and a FF of 0.52 compared to CuS 50 min, CuS 70 min, and a Pt CE. The superior performance of the 60 min sample is due to the greater electrocatalytic activity and low charge transfer resistance at the interface of the CE and the polysulfide electrolyte. The concentration of Cu/S also had an important role in the formation of the CuS nanoplatelet structures. The optical bandgaps for the CuS with different morphologies were measured to be in the range of 1.98-2.28 eV. This improved photovoltaic performance is mainly attributed to the greater number of active reaction sites created by the CuS layer on the FTO substrate, which results large specific surface, superior electrical conductivity, low charge transfer resistance, and faster electron transport in the presence of HCl. Cyclic voltammetry, electrochemical impedance spectroscopy and Tafel-polarization measurements were used to investigate the electrocatalytic activity of the CuS and Pt CEs. This synthetic procedure not only provides high electrocatalytic activity for QDSSCs but could also be a cost-effective way to fabricate flexible electrodes in dye-sensitized solar cells or supercapacitor applications.

Cost-benefit Analysis of Posaconazole Versus Fluconazole or Itraconazole as a Primary Antifungal Prophylaxis in High-risk Hematologic Patients: A Propensity Score-matched Analysis.

PURPOSE: Posaconazole is effective for the prophylaxis of invasive fungal infections (IFIs) in patients with acute myeloid leukemia or myelodysplastic syndrome during remission induction chemotherapy. However, a cost-benefit analysis of posaconazole versus fluconazole or itraconazole has not been conducted in Korea. METHODS: We retrospectively reviewed data for all consecutive patients who received primary antifungal prophylaxis during remission induction chemotherapy in our acute myeloid leukemia/myelodysplastic syndrome cohort from December 2010 to November 2013. Patient characteristics and factors known as a risk of IFI were matched with propensity score analysis. We evaluated the medical cost according to the prophylactic antifungal agents (posaconazole vs fluconazole/itraconazole), the development of breakthrough IFIs, and survival status after propensity score matching in a 1:1 ratio. FINDINGS: Of the 419 baseline patients, 100 patients in each group were analyzed after matching. A significant decrease was found in the development of breakthrough proven or probable IFIs (3.0% vs 14.0%; P = 0.009) and the rate of empirical antifungal therapy (EAFT) (12.0% vs 46.0%; P < 0.001) in the posaconazole group. Total in-hospital medical costs per patient were not statistically different between posaconazole and fluconazole/itraconazole prophylaxis. However, the daily medical cost was lower for posaconazole prophylaxis, resulting in a total daily cost savings of $72 (₩79,458) per patient (P = 0.002). In the cases of breakthrough proven/probable IFIs, EAFT, and in-hospital deaths, the total medical costs per patient were significantly higher than in nonproven/probable IFIs, non-EAFT, and in-hospital survivors, as much as $7,916 (₩8,700,758), $4605 (₩5,062,529), and $11,134 (₩12,238,422), respectively. Costs for the antifungal agent used in targeted or empirical therapy were lower in the posaconazole group, resulting in a savings of $697 (₩766,347) per patient (P < 0.001). IMPLICATIONS: Posaconazole appears to be cost beneficial for primary antifungal prophylaxis in high-risk patients with hematologic malignancy, at a single center, in Korea. Cost-benefit is closely related with clinical outcomes, including breakthrough IFI development, EAFT, and survival status.

Cost-effective and morphology controllable PVP based highly efficient CuS counter electrodes for high-efficiency quantum dot-sensitized solar cells.

Currently, copper sulfide (CuS) is the most commonly used counter electrode (CE) in high-efficiency quantum dot-sensitized solar cells (QDSSCs) because of its superior electrocatalytic activity in the presence of polysulfide electrolyte. For the first time, CuS thin films were prepared by a facile chemical bath deposition method with different concentrations of polyvinylpyrrolidone (PVP) and directly used as CEs in QDSSCs without any further post treatment. The quantum dot photoanode with the optimized 0.25 mM PVP-based CuS CE exhibits higher short circuit current density (Jsc), open circuit voltage (Voc), fill factor (FF), and power conversion efficiency (PCE) of 17.57 mA cm(-2), 0.578 V, 0.514, and 5.22%, respectively, which are much higher values than those of a bare CuS CE (Jsc: 12.36 mA cm(-2); Voc: 0.591 V; FF: 0.436; PCE: 3.18%) and Pt CE (Jsc: 11.25 mA cm(-2); Voc: 0.464 V; FF: 0.296; PCE: 1.54%) under one-sun illumination (AM 1.5 G, 100 mW cm(-2)). Moreover, the 0.25 mM PVP-based CuS CE produces a charge-transfer resistance of only 4.39 Ω with the aqueous polysulfide electrolyte commonly applied in QDSSCs. This value is several orders of magnitude lower than that of a typical Pt electrode (69.75 Ω) and bare CuS electrode (9.27 Ω). This enhancement is mainly attributed to the improved morphology of the 0.25 mM CuS CE with high catalytic activity, which plays a main role in the reduction processes of the oxidized polysulfide electrolyte, as well as the increased sulfur atomic percentage with Cu vacancies. Cyclic voltammetry, electrochemical impedance spectroscopy, and Tafel polarization were performed to study the underlying reasons behind the efficient CE performance.

Bone marrow plasma cell assessment before peripheral blood stem cell mobilization in patients with multiple myeloma undergoing autologous stem cell transplantation.

The current definition of complete response (CR) in multiple myeloma (MM) includes negative serum and urine immunofixation (IFE) tests and <5% bone marrow plasma cells (BMPCs). However, many studies of the prognostic impact of pretransplant response have not included BMPCs. We evaluated the prognostic impact of BMPC assessment before peripheral blood stem cell (PBSC) mobilization on subsequent transplant outcomes. BMPCs were assessed by CD138, kappa, and lambda immunostaining in 106 patients. After a median followup of 24.5 months, patients with <5% BMPCs had a significantly better progression-free survival (PFS) compared to those with ≥ 5% BMPCs (P = 0.005). Patients with <5% BMPCs + serologic CR showed superior PFS compared to those with <5% BMPCs + serologic non-CR (P = 0.050) or ≥ 5% BMPCs + serologic non-CR (P = 0.001). Interestingly, the prognostic impact of BMPCs was more apparent for patients who did not achieve a serologic CR (P = 0.042) compared to those with a serologic CR (P = 0.647). We concluded that IFE negativity and <5% BMPCs before PBSC mobilization were important factors to predict PFS in patients with MM undergoing ASCT. Particularly, a significant impact of <5% BMPCs was observed in patients who did not achieve IFE negativity.

Lymphocyte subset analysis for the assessment of treatment-related complications after autologous stem cell transplantation in multiple myeloma.

BACKGROUND AIMS: The aim of this study was to investigate the correlation between infused lymphocyte populations and lymphocyte subsets at engraftment, and the early clinical implications of lymphocyte subset recovery after autologous stem cell transplantation (ASCT) in multiple myeloma (MM). METHODS: We examined the lymphocyte populations of infused autografts and the lymphocyte subsets of peripheral blood at engraftment from 50 patients using flow cytometry. Each subset was grouped as low (below median) and high (above median) to examine the correlation with mucositis of grade 3 or more and the occurrence of infections and cytomegalovirus (CMV) reactivation. RESULTS: Using Spearman correlation coefficients, we found that cell doses of infused CD8(+) (P = 0.042) and CD19(+) cells (P = 0.044) were significantly associated with the absolute lymphocyte count (ALC) at engraftment. The dose of infused CD34(+) cells was not associated with the change of lymphocyte subsets except for an inverse correlation with CD4(+) cells (P = 0.006). After adjusting for potential variables in univariate analysis, multivariate analyzes revealed that the lower ratio of infused CD4(+) to CD8(+) cells (P = 0.030) was an independent factor for severe mucositis. Of lymphocyte subsets at engraftment, a higher frequency of CD3(+) (P = 0.024) and a lower frequency of CD56(+) (P = 0.020) were independent predictors for infections after engraftment. A higher frequency of CD8(+) cells (P = 0.041) and a lower ratio of CD4(+) to CD8(+) (P = 0.021) were independent predictors for CMV reactivation. CONCLUSIONS: Our data suggest that lymphocyte subset analysis of infused autograft and peripheral blood at engraftment may provide new predictors for early complications after ASCT in patient with MM.

Risk-adapted preemptive therapy for cytomegalovirus disease after allogeneic stem cell transplantation: a single-center experience in Korea.

Cytomegalovirus (CMV) remains a major cause of infection in recipients of hematopoietic stem cell transplants (HSCT) and results in significant mortality and morbidity. We present the results of CMV pp65 antigenemia-guided, risk-adapted preemptive therapy aimed at preventing CMV disease in allogeneic HSCT. Preemptive ganciclovir treatment was started when more than 5 CMV antigen-positive cells were detected in the low-risk group (with grade 0-I acute GVHD and matched related HSCT) and when any antigen-positive cells were seen in the high-risk group (with grade II-IV acute GVHD or matched unrelated HSCT). At least 1 episode of antigenemia was observed in 53 (59.6%) of 89 patients before day 100, and preemptive therapy was performed in 33 patients. CMV disease occurred in 6 patients (5 in the high-risk group and 1 in the low-risk group), and late CMV disease developed in 4 patients. Only 1 patient died of CMV pneumonitis before day 100. Neutropenia was observed in 51.5% of ganciclovir-treated patients, and coinfection/superinfection was observed in 42.4%. A strategy of ganciclovir treatment focusing on patients at higher risk could reduce the toxicity from the antiviral drug and be cost-effective. Extended surveillance for CMV disease using more sensitive diagnostic methods is necessary in high-risk patients.