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Hypercapnia occurs in ventilated infants even if tidal volume (VT) and minute ventilation (VE) are maintained. We hypothesised that increased physiological dead space (Vd,phys) caused decreased minute alveolar ventilation (VA; alveolar ventilation (VA) × respiratory rate) in well-ventilated infants with hypercapnia. We investigated the relationship between dead space and partial pressure of carbon dioxide (PaCO2) and assessed VA. Intubated infants (n = 33; mean birth weight, 2257 ± 641 g; mean gestational age, 35.0 ± 3.3 weeks) were enrolled. We performed volumetric capnography (Vcap), and calculated Vd,phys and VA when arterial blood sampling was necessary. PaCO2 was positively correlated with alveolar dead space (Vd,alv) (r = 0.54, p < 0.001) and Vd,phys (r = 0.48, p < 0.001), but not Fowler dead space (r = 0.14, p = 0.12). Normocapnia (82 measurements; 35 mmHg ≤ PaCO2 < 45 mmHg) and hypercapnia groups (57 measurements; 45 mmHg ≤ PaCO2) were classified. The hypercapnia group had higher Vd,phys (median 0.57 (IQR, 0.44-0.67)) than the normocapnia group (median Vd,phys/VT = 0.46 (IQR, 0.37-0.58)], with no difference in VT. The hypercapnia group had lower VA (123 (IQR, 87-166) ml/kg/min) than the normocapnia group (151 (IQR, 115-180) ml/kg/min), with no difference in VE.Conclusion: Reduction of VA in well-ventilated neonates induces hypercapnia, caused by an increase in Vd,phys. What is Known: • Volumetric capnography based on ventilator graphics and capnograms is a useful tool in determining physiological dead space of ventilated infants and investigating the cause of hypercapnia. What is New: • This study adds evidence that reduction in minute alveolar ventilation causes hypercapnia in ventilated neonates.

Background: We conducted active surveillance to elucidate the distribution of Streptococcus pneumoniae serotypes causing invasive pneumococcal disease (IPD) and clarified the genetic relatedness among the isolates in Kobe City, Japan. Methods: Forty-five IPD-causing S. pneumoniae strains were analyzed from March 2016 to May 2018 through active surveillance in Kobe City, Hyogo, Japan. Serotypes were determined by multiplex serotyping PCR and the Quellung reaction with pneumococcal antisera. Fourteen Sp12F strains were subjected to whole-genome sequencing (WGS). Results: Among 45 isolates, the most frequent serotypes were 12F (n = 14, 31%), 24F (n = 5, 11%), and 10A (n = 4, 9%). Multilocus sequence typing (MLST) analysis of 14 isolates of Sp12F divided them into ST4846 (n = 4) and ST6495 (n = 10). WGS showed clonality of the 10 isolates of ST6495, with only 13 single nucleotide polymorphisms in the genomes. Meanwhile, ST4846 strains in Kobe differed from only the outbreak strains of Sp12F ST4846 in Tsuruoka, Japan, reported on 2018. Conclusions: Serotype monitoring showed Sp12F to be the predominant serotype in Kobe, and WGS revealed the clonal spread of Sp12F ST6495 in this city. Thus, the spread of Sp12F could become a serious public health problem in Japan, warranting thorough monitoring in future. Keywords: Streptococcus pneumoniae, Serotype, Sp12F, Whole-genome sequencing, Single nucleotide polymorphism

Aim Intentional or unintentional acute drug poisoning occurs even in elderly people, but little is known about the factors influencing the intention to poisoning. A retrospective study was undertaken to describe the characteristics of acute drug poisoning in elderly people according to whether the poisoning was intentional or unintentional and the responsible agents. Methods The study was carried out in a single tertiary hospital in Japan. A total of 145 patients aged ≥65 years who were transferred by an ambulance service and were diagnosed with acute drug poisoning were included. Medical records were used to collect information on the intention behind poisoning and the responsible agents. Patients were divided into two groups according to whether they experienced intentional or unintentional poisoning and were further classified according to the responsible agent. Multivariable logistic regression models were used to estimate the association between hospitalization for acute drug poisoning and the use of benzodiazepine receptor agonists (BzRAs). Results Poisoning was unintentional in 102 (70.3%) patients and intentional in 43 (29.7%) patients. In total, 65 (44.8%) patients required hospitalization. Among patients in the unintentional poisoning group, those using non-BzRAs were more likely to be hospitalized than those using BzRAs (odds ratio, 6.64; 95% confidence interval, 2.56-17.22). The length of hospital stay was significantly longer in the unintentional poisoning group than in the intentional poisoning group (13.9 vs. 6.2 days; P = 0.013). Conclusions The proportion of unintentional poisoning in the elderly is high, and particularly with respect to poisoning with non-BzRAs, the hospitalization rates are high.

The risk of serious complication gradually increases as diabetes mellitus (DM) progresses. Thus, strategies for the prevention and delay of symptom progression are urgently needed. Previously, we synthesized zinc (Zn) complexes estimated to have a high bioavailability and evaluated their insulin-like anti-DM effects. However, in vivo studies of the effects of Zn compounds on the insulin signaling pathway and the molecular mechanisms underlying the anti-diabetic activities of Zn complexes were unresolved. In this study, we evaluated the effect of bis(maltolato)zinc(II) complex [Zn(mal)2] on male ICR mice (6-week-old) that received intraperitoneal (i.p.) injection of [Zn(mal)2]. The liver, skeletal muscle, and adipose tissues were collected from mice under anesthesia with isoflurane 40 or 90 min after i.p. injection. The [Zn(mal)2]-treatment did not affect Akt phosphorylation in the liver or skeletal muscle. In contrast, in adipose tissues, [Zn(mal)2]-treatment showed increased Akt phosphorylation at 40 min and 90 min after injection (p

Sunitinib has been shown to offer clinical benefits during the treatment of advanced renal cell carcinoma. However, molecular targeting drugs are expensive and can have a significant impact on medical expenses. The purpose of this study was to assess the cost-effectiveness of sunitinib as a first-line therapy compared with interferon-alpha (IFN-α) in metastatic renal cell carcinoma patients. A Markov model was used to show the clinical courses of patients with metastatic renal cell carcinoma who received sunitinib or IFN-α. The transition probabilities and utilities employed in this Markov model were derived from two sources. This study focused on the perspective of public healthcare payer, as only direct medical costs were estimated from the treatment schedule for metastatic renal cell cancer. In the cost-effectiveness analysis, outcomes were valued in terms of life years (LYs) and quality-adjusted life years (QALYs). We calculated the incremental cost-effectiveness ratio (ICER) during the cost-effectiveness analysis. The results were tested using Monte Carlo simulations. Sunitinib and IFN-α treatment resulted in LYs of 2.40 years and 2.03 years, QALYs of 1.58 and 1.25, and expected costs of 13,572,629 yen and 6,083,002 yen, respectively. As a result, the ICER associated with replacing IFN-α with sunitinib was 22,695,839 yen/QALYs. Our results suggest that compared with IFN-α, sunitinib prolongs LYs and QALYs, but the increases in quality achieved by sunitinib are more expensive than those produced by IFN-α.

Zinc (Zn) is a trace element with anti-diabetes mellitus (anti-DM) effects. Zn complexes exhibit stronger insulin-like activity than Zn ions. Bis(hinokitiolato)zinc complex ([Zn(hkt)2]) was recently reported to be a potent anti-DM candidate. We examined the effects of [Zn(hkt)2] on insulin resistance and pancreatic islet cells through in vivo long-term ingestion studies. In an in vivo study, we performed 4-month long-term [Zn(hkt)2] administration experiments in KK-Ay mice as a type 2 DM animal model. Ingestion of [Zn(hkt)2] resulted in lower blood glucose levels compared with the non-treated KK-Ay mice (control group). Additionally, [Zn(hkt)2] treatment decreased plasma insulin concentration compared with that of the non-treated KK-Ay group. [Zn(hkt)2] treatment resulted in a significant suppression of islet cell enlargement and a significantly decreased number of insulin-positive cells compared with the non-treated KK-Ay control group. The [Zn(hkt)2] treatment group showed the increasing tendency in the amount of Zn levels in peripheral organs; liver, muscle, adipose, and pancreas, compared with the non-treated KK-Ay control group. However, the Zn level in the pancreas of the [Zn(hkt)2] treatment group did not show the significant increase compared with the non-treated KK-Ay control group. This accumulation of Zn in pancreas suggested that [Zn(hkt)2] mainly effects on the peripheral tissue, and [Zn(hkt)2] has the less effect on the pancreas directly. Thus, we concluded that [Zn(hkt)2] exerted the main effect on peripheral organs by ameliorating insulin resistance.

In the present study, we investigated the effects of exercise intended to prevent or treat lifestyle-related diseases on the glucose tolerance, insulin level, lactic acid utilization, muscle glycogen synthesis, hepatic and renal oxidative stress, hepatic selenoprotein P and biological trace element levels in organs of obese, glucose-intolerant rats. We fed normal, healthy rats a 20% casein diet while the glucose-intolerant, obese rats received a high-fructose diet. They were forced to run for one hour per day, six days per week, for ten weeks. Exercise reduced visceral fat and ameliorated glucose tolerance in the high-fructose group, lowered blood lactic acid levels, improved lactic acid usage efficiency, and increased oxidative stress and hepatic levels of Mn, Fe, Cu, and Zn in the normal and high-fructose groups. Additionally, exercise significantly upregulated hepatic selenoprotein P expression in both groups, however, its effect was remarkable in healthy group. On the other hand, muscle glycogen synthesis was not markedly enhanced in high-fructose-diet rats but in normal-diet rats in response to exercise. It is concluded that exercise conditions rather than exercise load must be customized and optimized for each health and disease states in advance before starting exercise training intended to prevent or treat lifestyle-related diseases.

A 14-year-old girl was referred to our department because of headache and visual impairment following the resection of recurrent cardiac myxoma. Head magnetic resonance imaging (MRI) scan detected an intra- and supra-sellar tumor. Moreover, the patient showed the presence of spotty skin pigmentations on her cheeks and lower lip. Blood examination revealed hypothyrotropinemia, and ultrasonography results revealed multiple thyroid nodules. She was diagnosed with Carney complex (CNC). Her pituitary tumor was suspected as growth hormone (GH)-secreting adenoma, because overgrowth was observed in the patient. However, biochemical examinations, including oral glucose tolerance test, failed to show the characteristic findings of GH-secreting adenoma. In contrast, insulin tolerance test showed GH deficiency. Her visual impairment improved without performing decompression surgery, and the tumor size decreased, as per the MRI findings. Based on clinical course, the patient was diagnosed with pituitary apoplexy in pituitary adenoma, following which she was discharged. At 3 months after discharge, thyrotropin-releasing hormone loading test performed revealed low thyrotropin-stimulating hormone and thyroid hormone levels, and the patient was in a depressed mood. Therefore, l-T4 replacement was initiated, following which her GH secretory capacity gradually improved. Here, we report, to the best of our knowledge, the first case of a patient with pituitary apoplexy in CNC. Such condition must be identified in young patients with recurrent cardiac myxoma, and examinations, such as head MRI, must be performed.

Objective Constipation negatively affects the quality of life by causing abdominal pain and distention. It is common among women and older individuals and frequently encountered in Japan. Coffee and soda ingestion promote gastrointestinal motility; therefore, both liquids are considered effective for alleviating constipation. While invasive approaches or large-scale equipment, such as magnetic resonance imaging and radiography, are needed to confirm their effect on gastrointestinal motility, bowel sound (BS) assessment can be easily and non-invasively performed using low-cost equipment. Differently to coffee, soda contains carbon dioxide and its ingestion may therefore increase gastrointestinal gas and in turn affect the BS generation process. However, the relationship between gastrointestinal gas and acoustic BS features has not yet been investigated. This study aimed to investigate the effect of increased gastrointestinal gas resulting from soda ingestion on the acoustic BS features extracted using a newly improved BS detection technique. Methods We used BS-based acoustic features, including number of BSs detected/min, BS length, and BS power, extracted via an artificial neural network-based BS detection approach, which was developed in a previous study and here improved. Results BSs detected/min and BS length significantly increased after, compared with before, coffee and soda intake. Furthermore, we found that longer BSs (>128 ms) were more significantly associated with soda (200 mL) intake than with coffee (200 mL) intake. Conclusions BS length and BSs detected/min can sensitively evaluate gastrointestinal motility, and BS length may reflect information on gastrointestinal gas in addition to gastrointestinal motility. Recording of BS-based acoustic features is a non-invasive and convenient means to evaluate the effects of liquid intake on gastrointestinal motility in patients with functional gastrointestinal disorders.

α-Amino acids and their derivatives are important and essential resources for human life such as foods, medical drugs, cosmetics and so on. The classical Strecker α-amino acids synthesis is a longstanding method although this classical method requires highly toxic cyanide reagents as the carbon source and these regents are unfavorable from the viewpoint of green chemistry. Carbon dioxide, on the other hand, would also be an abundant and favorable carbon source. There are several examples of synthesis of α-amino acids by direct carboxylation of imines with carbon dioxide. Because of chemical stability of carbon dioxide, this direct carboxylation requires stoichiometric quantities of hard to handle reagents such as magnesium metal and organotins under severe or harsh reaction conditions. In this context, electrochemical carboxylation represents an attractive alternative to conventional chemical carboxylation methods because it is possible to readily generate highly reactive carbon nucleophiles that immediately react with carbon dioxide without handling sensitive, expensive, and toxic reagent. However, metal ions derived from sacrificial anode such as magnesium and aluminium anodes are generally required to stabilize the unstable carboxylate ions. A flow microreactor enables fast generation and consumption of the unstable species. Therefore, it is attractive reaction system for electrochemical carboxylation involving unstable carboxylate ions without sacrificial anodes. Moreover, the electrochemical synthesis with a flow microreactor offers operational advantages over batch processes. Under these backgrounds, we have demonstrated the electrochemical carboxylation of in situ generated imines in a continuous flow system. The present method enables an efficient synthesis for N-phenylphenylglycine derivatives using easily available aldehydes, amines, and CO2 without the use of toxic metal reagents and sacrificial anode. As shown in Figure 1, the imine was smoothly generated from aldehyde and amine by dehydration using MS4A desiccant in a flow operation, and then reacted with carbon dioxide in the electrochemical flow microreactor to afford N-phenylphenylglycine derivatives with high to moderate yields. Figure 1

Aims Previous studies reported the anti-diabetic effects of α-lipoic acid (αLA) isomers: racemic-αLA, R-αLA, or S-αLA. Previously, we examined the anti-diabetic effects of αLA administered as a food additive, but were unable to demonstrate the differences among different isomers. In this study, αLAs were complexed with γ-cyclodextrin (γCD) for the stability. We then investigated the anti-diabetic effects of racemic-, R-, and S-αLA/γCDs in KKA y mice. Main methods Male type 2 diabetic KKA y mice were divided into 5 groups, and fed either a high-fat-diet (HFD), HFD supplemented with γCD, or HFD supplemented with racemic-αLA/γCD, R-αLA/γCD, or S-αLA/γCD for 4 weeks. At the end of the feeding period, HbA1c and adiponectin levels were measured, PPARγ2 mRNA expression levels were assessed in adipose tissues using real-time PCR, and AMP-activated protein kinase (AMPK) phosphorylation levels were evaluated in the liver by Western blotting. Key findings The anti-diabetic effects of αLA; the isomeric compounds racemic-, R-, and S-αLA/γCD were investigated using a male type 2 diabetic KKA y mouse model. Significant differences were observed in HbA1c and plasma adiponectin levels between R-αLA/γCD-treated mice and control mice. PPARγ2 mRNA expression levels were slightly higher in racemic- and R-αLA/γCD-treated mice. Moreover, AMPK phosphorylation levels were elevated in racemic-αLA/γCD- and R-αLA/γCD-treated mice, but remained unchanged in S-αLA/γCD-treated mice. Significance These results suggested that the stereoisomerism mediates a difference in the anti-diabetic effects of racemic-, R-, and S-αLA/γCDs. Furthermore, the anti-diabetic mechanism of αLA/γCD action may be attributed to the activation of AMPK in the liver.

Botulinum toxin A (BTXA) can disrupt the neuromuscular and autonomic functions. We herein report a case of autonomic system dysfunction that manifested as Takotsubo-like myocardial dysfunction in a patient with botulism. Takotsubo syndrome results in acute cardiac insufficiency, another fatal complication of botulism in addition to respiratory muscle paralysis, particularly in patients with cardiovascular disease.

Background/aim Metabolic syndrome-induced lifestyle-related diseases include diabetes mellitus (DM) and hypertension, and Zn-based compounds have effects on DM. We aimed to investigate the ameliorating effects of bis(hinokitiolato)Zn, [Zn(hkt)2] on lipid metabolism in the liver and kidney, histopathologically. Materials and methods We used a high-fat diet (HFD)-fed C57BL/6J mouse model and administered a diet containing 10-20 mg Zn/kg body weight (BW) or 20 mg pioglitazone/kg BW as the positive control. After the treatments, we collected blood, liver, and kidney samples and morphologically evaluated the mouse organs for fat accumulation. Results After a 4-month HFD administration, ectopic fat deposition was detected in the liver and kidney. Furthermore, Zn accumulation in the liver and kidney increased following [Zn(hkt)2] treatment, that reduced lipid accumulations and lipid toxicity in these tissues. Conclusion The results of this study suggest that [Zn(hkt)2] could be a novel anti-dyslipidaemia compound for treating diet-induced obesity.

Radiotherapy (RT) and chemoradiotherapy (CRT) is widely accepted as the standard treatment for head and neck cancer (HNC). Oral mucositis (OM) often develops as an adverse reaction in HNC patients that receive RT or CRT involving S-1. However, little is known about the risk factors for OM in HNC patients. We retrospectively evaluated patients' pre-treatment clinical data in order to identify the risk factors for severe OM in HNC patients that are treated with RT or CRT involving S-1. We analyzed the cases of 129 patients who received RT or CRT involving S-1 for HNC. The endpoint of the survey was the occurrence of severe OM (≥grade 2). Risk factors that were significantly related to severe OM were identified using logistic regression analysis. The patients' mean age was 69.3±10.1 years, and 118 (92%) of the patients were male. The primary tumor was located in the oropharynx in 21.7% of cases. Severe OM occurred in 85.0% of cases. In the univariate analysis, the following variables were found to be associated with severe OM: age, the type of radiotherapy, disease stage, and chemotherapy. In the multivariate analysis, the location of the primary tumor and chemotherapy were identified as significant risk factors that contributed independently to the risk of severe OM (p

In recent years, the number of patients suffering from diabetes mellitus has been increasing worldwide. In particular, type 2 diabetes mellitus, a lifestyle-related disease, is recognized as a serious disease with various complications. Many types of pharmaceutics or specific health foods have been used for the management of diabetes mellitus. At the same time, the relationship between diabetes mellitus and α-lipoic acid has been recognized for many years. In this study, we found that the α-lipoic acid γ-cyclodextrin complex exhibited an HbA1c lowering effect for treating type 2 diabetes mellitus in animal models. Moreover, in this study, we investigated the activation of phosphorylation of AMP-activated protein kinase, which plays a role in cellular energy homeostasis, in the liver of KKA(y) mice by using α-lipoic acid and the α-lipoic acid γ-cyclodextrin complex. Our results show that the α-lipoic acid γ-cyclodextrin complex strongly induced the phosphorylation of AMP-activated protein kinase. Thus, we concluded that intake of the α-lipoic acid γ-cyclodextrin complex exerted an antidiabetic effect by suppressing the elevation of postprandial hyperglycemia as well as doing exercise.

洗浄・置換血小板（W/R-PC）は血小板製剤による輸血副作用を防止する目的で調製されている．我々は血小板製剤の洗浄および置換には臨床使用が認められた輸液および電解質溶液を混和させることで製造される洗浄置換液（M-sol）を用いている．M-solの製造はクリーンルームにて用時行っているが，本研究では以下の二点を中心に製造方法の改良を行った．1）除菌フィルター付バッグを使用することで，クリーンルーム外での製造を可能とした．2）調製したM-solはアルミ蒸着袋に真空保存することで，pHを一定に保つことが可能となり最短でも1年間の保存が可能であった．さらに我々はW/R-PC 1バッグ分のM-solを製造する方法（単品式）と一度に多量のM-solを製造する方法（多量式）を考案し，両法によるM-solに差がないことを確認した．真空包装したM-solは30℃で1年間保存してもpHに変化はなかった．さらに，製造直後のM-solと長期保存したM-solにより調製された置換PC（R-PC）には血小板機能に差はみられなかった．また，長距離輸送したM-solを用いて調製した洗浄PC（W-PC）についても，血小板への影響はみられなかった．改良された方法ではM-solの製造時間に制限されずにW/R-PCの調製が可能となるであろう．

血小板輸血による副作用を予防する目的で血漿を除去した洗浄·置換血小板（Washed and/or replaced platelet concentrates，W/R-PC）が調製されている．調製法としては2つの方法が主に使われているが両者を比較した報告はない．そこで，成分採血由来の血小板（Plasma-PC）を洗浄置換·保存液M-solで洗浄し，M-solに再浮遊する方法（洗浄置換法WR-method）とplasma-PC中の血漿を遠心後M-solに置換する方法（置換法R-method）について，各W/R-PCの血小板機能や輸血効果を比較検討した． 各方法で調製されたW/R-PCの血小板回収率や残存血漿蛋白，in vitroにおける性状（pH，凝集能，%HSR，P-セレクチン陽性率，血小板形態）を調べ，アナフィラキシーショックなどの重篤な副作用がみられた患者を対象に，WR-methodによるPC（WR-PC）を75バッグ（患者: 6名），R-methodによるPC（R-PC）を31バッグ（患者: 4名）に投与して，補正血小板増加数（CCI）及び副作用の予防効果を比較した． WR-PCとR-PCの血小板回収率はそれぞれ90.5±1.4%と89.5±1.8%で有意差はなく，血漿蛋白除去率（残存蛋白量）は，WR-PCでは96.9±0.7%（428±95mg）で，R-PCの95.4±0.9%（627±130mg）より有意に高かった（各n=7）．48時間の保存期間中（PC採血後5日目），in vitroの性状はpH以外の項目に差はみられなかった．Plasma-PCは保存と共にpHが低下し，24時間後には7.05±0.04であったのに対し，W/R-PCは何れも調製直後に一時的なpHの低下が認められたが，WR-PCでは7.37±0.03，R-PCは7.40±0.02に上昇した．輸血24時間後のCCI（×104/ul）は，WR-PCが1.53±0.82（n=51），R-PCが1.59±0.78（n=18）と良好であり，何れのW/R-PCについても，輸血後の副作用の発生はなく，副作用予防効果が示された．また，何れも有害事象は認められなかった． WR-PCとR-PCは，血小板機能や輸血効果について，同等である事が確認されたが，R-methodは調製工程が少なく，より簡便であるため，PCの血漿除去法として推奨される．

Fgf19 is known to be expressed in the developing chicken eye but its functions during retinal development have remained elusive. Since Fgf19 is expressed in the dorsal portion of the optic cup, it is intriguing to know whether FGF19 is required for expression of dorso-ventral morphogenetic genes in the eye. To clarify this, expression patterns of Tbx5 and Vax were examined in the developing eye after in ovo RNA interference targeted against Fgf19. Quantitative polymerase chain reaction (PCR) analysis showed that the short-hairpin RNAs (shRNAs) targeted against Fgf19 could reduce its expression in the eye to less than 50% of a relative amount of mRNA, compared with contralateral or untreated control eyes. However, no obvious alteration in expression domains of Tbx5 or Vax was observed. Misexpression of Tbx5 or Tbx5-RNAi did not alter the Fgf19 expression either. Furthermore, although Fgf19 is expressed in the central retina before neurogenesis occurs, β3-tubulin, a marker for early retinal differentiation was still detected in the central retina after knockdown of Fgf19. Thus, knockdown of Fgf19 supports no obvious regulations between Fgf19 and Tbx5, or exhibits no phenotypes that perturb early retinal differentiation.

siVirus (http://siVirus.RNAi.jp/) is a web-based online software system that provides efficient short interfering RNA (siRNA) design for antiviral RNA interference (RNAi). siVirus searches for functional, off-target minimized siRNAs targeting highly conserved regions of divergent viral sequences. These siRNAs are expected to resist viral mutational escape, since their highly conserved targets likely contain structurally/functionally constrained elements. siVirus will be a useful tool for designing optimal siRNAs targeting highly divergent pathogens, including human immunodeficiency virus (HIV), hepatitis C virus (HCV), influenza virus and SARS coronavirus, all of which pose enormous threats to global human health.

With melanoma, as with many other malignancies, aberrant transcriptional repression is a hallmark of refractory cancer. To restore gene expression, use of a histone deacetylase inhibitor (HDACi) is expected to be effective. Our recent DNA micro-array analysis showed that the HDACi depsipeptide (FK228) significantly enhances gp100 antigen expression. Herein, we demonstrate that depsipeptide promotes tumor-specific T-cell-mediated killing of B16/F10 murine melanoma cells. First, by a quantitative assay of caspase-3/7 activity, a sublethal dose of depsipeptide was determined (ED50: 5 nM), in which p21(Waf1/Cip1) and Fas were sufficiently evoked concomitantly with histone H3 acetylation. Second, the sublethal dose of depsipeptide treatment with either a recombinant Fas ligand or tumor-specific T cells synergistically enhanced apoptotic cell death in B16/F10 cells in vitro. Furthermore, we found that depsipeptide increased levels of perforin in T cells. Finally, in vivo metastatic growth of B16/F10 in the lung was significantly inhibited by a combination of depsipeptide treatment and immune cell adoptive transfer from immunized mice using irradiated B16 cells and gp100-specific (Pmel-1) TCR transgenic mice (P

We propose rational designing of antiviral short-interfering RNA (siRNA) targeting highly divergent HIV-1. In this study, conserved regions within HIV-1 genomes were identified through an exhaustive computational analysis, and the functionality of siRNAs targeting the highest possible conserved regions was validated. We present several promising antiviral siRNA candidates that effectively inhibited multiple subtypes of HIV-1 by targeting the best conserved regions in pandemic HIV-1 group M strains.

To avoid long-double-stranded-RNA-dependent interferon response, short-interfering RNAs (siRNAs) are widely used for RNA interference (RNAi) in mammalian cells. siRNA-based RNAi, however, may not be readily available for the large-scale gene silencing essential for systematic functional genomics, because only a limited fraction of siRNAs is capable of inducing effective mammalian RNAi. siRNAs correctly designed for the knockdown of a particular gene may also destroy the functions of unrelated genes. Here, we describe algorithms by which these serious setbacks can be eliminated in mammalian functional genomics using RNAi and a Web-based online software system for computing highly functional siRNA sequences with maximal target-specificity in mammalian RNAi.