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Given the worldwide burden of neglected tropical diseases, there is ongoing need to develop novel anthelmintic agents to strengthen the pipeline of drugs to combat these burdensome infections. Many diseases caused by parasitic flatworms are treated using the anthelmintic drug praziquantel (PZQ), employed for decades as the key clinical agent to treat schistosomiasis. PZQ activates a flatworm transient receptor potential (TRP) channel within the melastatin family (TRPMPZQ) to mediate sustained Ca2+ influx and worm paralysis. As a druggable target present in many parasitic flatworms, TRPMPZQ is a promising target for a target-based screening campaign with the goal of discovering novel regulators of this channel complex. Here, we have optimized methods to miniaturize a Ca2+-based reporter assay for Schistosoma mansoni TRPMPZQ (Sm.TRPMPZQ) activity enabling a high throughput screening (HTS) approach. This methodology will enable further HTS efforts against Sm.TRPMPZQ as well as other flatworm ion channels. A pilot screen of ~16,000 compounds yielded a novel activator of Sm.TRPMPZQ, and numerous potential blockers. The new activator of Sm.TRPMPZQ represented a distinct chemotype to PZQ, but is a known chemical entity previously identified by phenotypic screening. The fact that a compound prioritized from a phenotypic screening campaign is revealed to act, like PZQ, as an Sm.TRPMPZQ agonist underscores the validity of TRPMPZQ as a druggable target for antischistosomal ligands., Author summary The drug praziquantel is used to treat diseases caused by parasitic flatworms. Praziquantel is an old drug, and there is a need to identify novel treatments that retain desirable features and improve weaknesses in the mode of PZQ action. One way to do this is to identify new drugs that exploit vulnerabilities in the same drug target but work in slightly differently ways. Here, we have optimized high throughput screening methods to pharmacologically profile a parasitic flatworm ion channel targeted by PZQ. We have identified several new chemical structures that interact with this channel complex. These ligands provide new opportunity for developing tools to manipulate flatworm biology and potentially new trajectories for anthelmintic drug development.

Curcumin, a compound found in Indian yellow curry, is known to possess various biological activities, including anti-oxidant, anti-inflammatory, and anti-cancer activities. Cur2004-8 is a synthetic curcumin derivative having symmetrical bis-alkynyl pyridines that shows a strong anti-angiogenic activity. In the present study, we examined the effect of dietary supplementation with Cur2004-8 on response to environmental stresses and aging using Caenorhabditis elegans as a model system. Dietary intervention with Cur2004-8 significantly increased resistance of C. elegans to oxidative stress. Its anti-oxidative-stress effect was greater than curcumin. However, response of C. elegans to heat stress or ultraviolet irradiation was not significantly affected by Cur2004-8. Next, we examined the effect of Cur2004-8 on aging. Cur2004-8 significantly extended both mean and maximum lifespan, accompanying a shift in time-course distribution of progeny production. Age-related decline in motility was also delayed by supplementation with Cur2004-8. In addition, Cur2004-8 prevented amyloid-beta-induced toxicity in Alzheimer's disease model animals which required a forkhead box (FOXO) transcription factor DAF-16. Dietary supplementation with Cur2004-8 also reversed the increase of mortality observed in worms treated with high-glucose-diet. These results suggest that Cur2004-8 has higher anti-oxidant and anti-aging activities than curcumin. It can be used for the development of novel anti-aging product.

Phosphatidylcholine is one of the major phospholipids comprising cellular membrane and is known to have several health-promoting activities, including the improvement of brain function and liver repair. In this paper, we examine thein vivoeffect of dietary supplementation with phosphatidylcholine on the response to environmental stressors and aging inC. elegans. Treatment with phosphatidylcholine significantly increased the survival of worms under oxidative stress conditions. However, there was no significant difference in response to stresses caused by heat shock or ultraviolet irradiation. Oxidative stress is believed to be one of the major causal factors of aging. Then, we examined the effect of phosphatidylcholine on lifespan and age-related physiological changes. Phosphatidylcholine showed a lifespan-extending effect and a reduction in fertility, possibly as a tradeoff for long lifespan. Age-related decline of motility was also significantly delayed by supplementation with phosphatidylcholine. Interestingly, the expressions of well-known longevity-assuring genes,hsp-16.2andsod-3, were significantly upregulated by dietary intervention with phosphatidylcholine. DAF-16, a transcription factor modulating stress response genes, was accumulated in the nucleus by phosphatidylcholine treatment. Increase of the ROS level with phosphatidylcholine suggests that the antioxidant and lifespan-extending effects are due to the hormetic effect of phosphatidylcholine. Phosphatidylcholine also showed a protective effect against amyloid beta-induced toxicity in Alzheimer’s disease model animals. Experiments with long-lived mutants revealed that the lifespan-extending effect of phosphatidylcholine specifically overlapped with that of reduced insulin/IGF-1-like signaling and required DAF-16. These findings showed the antioxidant and antiaging activities of phosphatidylcholine for the first timein vivo. Further studies focusing on the identification of underlying cellular mechanisms involved in the antiaging effect will increase the possibility of using phosphatidylcholine for the development of antiaging therapeutics.

Praziquantel (PZQ) is an essential medicine for treating parasitic flatworm infections such as schistosomiasis, which afflicts over 250 million people. However, PZQ is not universally effective, lacking activity against the liver fluke Fasciola. The reason for this insensitivity is unclear, as the mechanism of PZQ action is unknown. Here, we show PZQ activates a transient receptor potential melastatin ion channel (TRPMPZQ) in schistosomes by engaging a hydrophobic ligand binding pocket within the voltage-sensor like domain to cause Ca2+ entry and worm paralysis. PZQ activates TRPMPZQ homologues in other PZQ-sensitive flukes, but not Fasciola. However, a single amino acid change in the Fasciola TRPMPZQ binding pocket, to mimic schistosome TRPMPZQ, confers PZQ sensitivity. After decades of clinical use, the basis of PZQ action at a druggable TRP channel is resolved.

Cestodes are platyhelminth parasites with a wide range of hosts that cause neglected diseases. Neurotransmitter signaling is of critical importance for these parasites which lack circulatory, respiratory and digestive systems. For example, serotonin (5-HT) and serotonergic G-protein coupled receptors (5-HT GPCRs) play major roles in cestode motility, development and reproduction. In previous work, we deorphanized a group of 5-HT7 type GPCRs from cestodes. However, little is known about another type of 5-HT GPCR, the 5-HT1 clade, which has been studied in several invertebrate phyla but not in platyhelminthes. Three putative 5-HT GPCRs from Echinococcus canadensis, Mesocestoides vogae (syn. M. corti) and Hymenolepis microstoma were cloned, sequenced and bioinformatically analyzed. Evidence grouped these new sequences within the 5-HT1 clade of GPCRs but differences in highly conserved GPCR motifs were observed. Transcriptomic analysis, heterologous expression and immunolocalization studies were performed to characterize the E. canadensis receptor, called Eca-5-HT1a. Functional heterologous expression studies showed that Eca-5-HT1a is highly specific for serotonin. 5-Methoxytryptamine and α-methylserotonin, both known 5-HT GPCR agonists, give stimulatory responses whereas methysergide, a known 5-HT GPCR ligand, give an antagonist response in Eca-5-HT1a. Mutants obtained by the substitution of key predicted residues resulted in severe impairment of receptor activity, confirming that indeed, these residues have important roles in receptor function. Immunolocalization studies on the protoscolex stage from E. canadensis, showed that Eca-5-HT1a is localized in branched fibers which correspond to the nervous system of the parasite. The patterns of immunoreactive fibers for Eca-5-HT1a and for serotonin were intimately intertwined but not identical, suggesting that they are two separate groups of fibers. These data provide the first functional, pharmacological and localization report of a serotonergic receptor that putatively belongs to the 5-HT1 type of GPCRs in cestodes. The serotonergic GPCR characterized here may represent a new target for antiparasitic intervention.

The probability of objects fabricated by three-dimensional (3D) printing exhibiting local defects is higher than that detected in products of conventional casting-based manufacturing. Multistep layer-by-layer procedures in additive manufacturing are the main reason. Light intensity and/or penetration depth, inhomogeneity of components, and variations in nozzle temperature are factors that create local defects. Defect regions are sources of toxic component release, but methods to identify them in printed materials have not been reported. Existing assays for evaluating material toxicity are based on extraction, and these toxicological assays use living creatures to passively detect harmful agents in extracted solutions. Thus, the development of an active system for identifying sites of toxicity sources is a critical and urgent issue in 3D printing technologies. Herein, we introduce an animal model system, C. elegans, for toxicity evaluation. C. elegans crawls toward safe regions but avoids toxically dangerous areas. The ‘sensing’ and ‘locomotion’ abilities of C. elegans are unparalleled among existing underwater and animal models, providing immediate indications to help find toxicity source sites.

The effects of antioxidants on lifespan have been widely studied. Our previous study showed supplementation with N-acetyl-l-cysteine (NAC) extends the lifespan of Caenorhabditis elegans. Here we aimed to determine the lifespan-extending mechanism involved with NAC and the effect of NAC on Alzheimer’s disease (AD). NAC further increased the lifespan of age-1 and clk-1 mutants, which have increased lifespan owing to reduced insulin/IGF-1-like signaling and mitochondrial function, respectively. There was no additional lifespan extension in eat-2 background, a genetic model of dietary restriction (DR), by NAC. Gene knockdown experiments revealed that the effect of NAC is not dependent on SKN-1, a protein-sensing DR status, whereas DAF-16, a transcription factor regulating stress-responsive genes, is required for lifespan extension by NAC. NAC delayed paralysis caused by amyloid beta. Our results show that NAC mimics the effect of DR on lifespan, possibly through the induction of DAF-16 nuclear localization and may retard the incidence of AD.

Triptolide is a major active compound found in Tripterygium wilfordii., also known as Thunder God Vine. Triptolide has been shown to have anti-inflammatory and anticancer activities. In this study, we examined the effect of dietary supplementation with triptolide on response to environmental stressors and lifespan in vivo using Caenorhabditis elegans as a model system. Treatment with 50 mg/L of triptolide in the growth media increased resistance to oxidative stress and reduced the generation of intracellular reactive oxygen species. We also observed a lifespan-extending activity for triptolide. Both mean and maximum lifespans were significantly increased by supplementation with triptolide. Response to other environmental stressors was modulated by triptolide. The survival after heat shock or UV irradiation was markedly increased in worms treated with triptolide. Unlike many lifespan-extending genetic or nutritional interventions, the longevity phenotype conferred by triptolide did not have the trade-off of a reduction in fertility or a delay in the gravid period. The expressions of hsp-16.2 and sod-3, known to positively correlate with a long lifespan, were significantly upregulated by supplementation with triptolide. These findings suggest that triptolide can exhibit antistress and lifespan-extending effects in vivo, possibly through its antioxidant activity and support the free radical theory of aging, which emphasizes the causative role of oxidative stress in aging.

Dietary restriction has been shown to have a lifespan-extending effect in various model organisms, from yeast to mice. The nematode C. elegans is widely used as a model organism for researches on dietary restriction due to various dietary restriction methods developed in C. elegans. Previous study showed that coelomocyte-specific cup-4 and cup-5 genes are necessary for dietary-restriction-induced longevity in C. elegans. We examined the effect of coelomocyte knockout on lifespan extension by different methods of dietary restriction. The extension of both mean and maximum lifespan by bacterial dilution was completely abolished by genetic ablation of coelomocytes. Knockout of coelomocytes also blocked the lifespan extension conferred by food deprivation or removal of peptone in culture media. These findings indicate that coelomocytes are required for lifespan extension by different methods of dietary restriction, and suggest that the activation of coelomocytes may be a common underlying mechanism involved in dietary-restriction-induced longevity in C. elegans.

The insulin/insulin-like growth factor (IGF)-1 signaling (IIS) pathway is a conserved lifespan-modulating genetic pathway. Many genes involved in lifespan extension associated with decreased signaling of the IIS pathway have been identified. In the present study, we found a novel gene required for the effect of the IIS pathway on the stress response and aging in C. elegans . Receptor mediated endocytosis (RME)-1 is expressed ubiquitously and known to be involved in cellular endocytic transport. Knockdown of rme-1 abolished the lifespan-extending effect caused by decreased IIS. In addition, resistance to oxidative stress, heat shock and ultraviolet irradiation were significantly decreased when the expression of rme-1 was blocked. The delayed age-related decline in motility observed in age-1 mutants with defects in the IIS pathway was also modulated by RME-1. The expression of sod-3 , which is positively correlated with the remaining lifespan of an individual, was decreased by rme-1 knockdown. Our study demonstrates that RME-1 is required for the anti-aging effect associated with decreased IIS. We suggest that endocytic transport could be one underlying mechanisms for longevity via the IIS pathway. https://doi.org/10.2298/ABS160510115K Received: May 10, 2016; Revised: June 10, 2016; Accepted: June 10, 2016; Published online: October 31, 2016 How to cite: Kim CK, Park SK. RME-1 is required for lifespan extension and increased resistance to stresses associated with decreased insulin/IGF-1-like signaling in Caenorhabditis elegans. Arch Biol Sci. 2017;69(3):417-25.

Oral propranolol has become first-line treatment for infantile hemangiomas (IHs). This study focused on identifying cytokines related to the biology of IH and early regression indicators of IH after propranolol treatment.For inclusion, the patients had to be aged less than 1 year and have an IH with a largest diameter ≥2 cm. Patients were scheduled to receive 1 year of propranolol treatment. Serum cytokines involved in angiogenesis, vasculogenesis, and/or chronic inflammation were analyzed at 0, 1, and/or 12 months after treatment using Multiplex Luminex assays.Among the 49 evaluable patients, 33 completed the 1-year treatment: 16 showed excellent response and 12 had good response to propranolol. Significant decreases in serum MMP-2, bFGF, VEGF-α, and MCP-1 levels were observed after 1 year of treatment compared to pretreatment values. The maximal diameters of the lesions significantly correlated with pretreatment serum VEGF-α, bFGF, and MMP-9. Patients with higher bFGF and VEGF levels showed better response to propranolol at 1 year.MMP-2, VEGF-α, bFGF, and MCP-1 may involve in the biology of IH and their downregulation may be associated with involution processes of IH. Pretreatment bFGF and VEGF could be novel biomarkers for predicting response to propranolol.We found that decreases in the concentrations of MMP-2, bFGF, VEGF, and MCP-1 were associated with regression of the hemangioma, which indicates that one of the mechanisms of propranolol in the treatment of proliferative hemangiomas may involve downregulation of those cytokines. Patients with higher bFGF and VEGF levels showed better response to propranolol at 1 year. Importantly, serum bFGF higher than 37.07 pg/mL may predict an excellent response to propranolol. Therefore, along with the patient's age and the size and visual characteristics of the lesion, bFGF levels could help determine the viability of propranolol use in the treatment of IHs. Our study represented extensive serum profiling in IH, reporting the indicators and molecules clearly related to IH regression with propranolol treatment. The authors believe that monitoring serum cytokines, including MMP-2, bFGF, VEGF, and MCP-1, in IH patients could be important, in addition to clinical follow-up, for determining when to start and end propranolol treatment.

Infections caused by parasitic flatworms impose a considerable worldwide health burden. One of the most impactful is schistosomiasis, a disease caused by parasitic blood flukes. Treatment of schistosomiasis has relied on a single drug – praziquantel (PZQ) – for decades. The utility of PZQ as an essential medication is, however, intertwined with a stark gap in our knowledge as to how this drug works. No flatworm target has been identified that readily explains how PZQ paralyzes and damages schistosomes. Recently, a schistosome ion channel was discovered that is activated by PZQ and displays characteristics which mirror key features of PZQ action on schistosomes. Here, the journey to discovery of this target, properties of this ion channel, and remaining questions are reviewed.

The anthelmintic drug praziquantel (PZQ) is used to treat schistosomiasis, a neglected tropical disease that affects over 200 million people worldwide. PZQ causes Ca2+ influx and spastic paralysis of adult worms and rapid vacuolization of the worm surface. However, the mechanism of action of PZQ remains unknown even after 40 years of clinical use. Here, we demonstrate that PZQ activates a schistosome transient receptor potential (TRP) channel, christened Sm.TRPMPZQ, present in parasitic schistosomes and other PZQ-sensitive parasites. Several properties of Sm.TRPMPZQ were consistent with known effects of PZQ on schistosomes, including (i) nanomolar sensitivity to PZQ; (ii) stereoselectivity toward (R)-PZQ; (iii) mediation of sustained Ca2+ signals in response to PZQ; and (iv) a pharmacological profile that mirrors the well-known effects of PZQ on muscle contraction and tegumental disruption. We anticipate that these findings will spur development of novel therapeutic interventions to manage schistosome infections and broader interest in PZQ, which is finally unmasked as a potent flatworm TRP channel activator.

Background Current diagnostic tests for hereditary spherocytosis (HS) focus on the detection of hemolysis or indirectly assessing defects of membrane protein, whereas direct methods to detect protein defects are complicated and difficult to implement. In the present study, we investigated the patterns of genetic variation associated with HS among patients clinically diagnosed with HS. Methods Multi-gene targeted sequencing of 43 genes (17 RBC membrane protein-encoding genes, 20 RBC enzyme-encoding genes, and six additional genes for the differential diagnosis) was performed using the Illumina HiSeq platform. Results Among 59 patients with HS, 50 (84.7%) had one or more significant variants in a RBC membrane protein-encoding genes. A total of 54 significant variants including 46 novel mutations were detected in six RBC membrane protein-encoding genes, with the highest number of variants found in SPTB (n = 28), and followed by ANK1 (n = 19), SLC4A1 (n = 3), SPTA1 (n = 2), EPB41 (n = 1), and EPB42 (n = 1). Concurrent mutations of genes encoding RBC enzymes (ALDOB, GAPDH, and GSR) were detected in three patients. UGT1A1 mutations were present in 24 patients (40.7%). Positive rate of osmotic fragility test was 86.8% among patients harboring HS-related gene mutations. Conclusions This constitutes the first large-scaled genetic study of Korean patients with HS. We demonstrated that multi-gene target sequencing is sensitive and feasible that can be used as a powerful tool for diagnosing HS. Considering the discrepancies of clinical and molecular diagnoses of HS, our findings suggest that molecular genetic analysis is required for accurate diagnosis of HS. Electronic supplementary material The online version of this article (10.1186/s13023-019-1070-0) contains supplementary material, which is available to authorized users.

The anthelmintic drug praziquantel (PZQ) is used to treat schistosomiasis, a neglected tropical disease that affects over 200 million people. The target of PZQ in the blood fluke responsible for this disease is unknown. Here, we demonstrate that PZQ activates a transient receptor potential (TRP) channel found in parasitic schistosomes and other PZQ-sensitive parasites.

It is suggested that oxidative stress induced by cellular reactive oxygen species is one of the major causal factors of aging. The effect of dietary supplementation of anti-oxidants on response to environmental stressors and lifespan has been studied in various model organisms. In the present study, we examine the effect of Artemisia annua extract on resistance to oxidative, heat, and ultraviolet stresses in the nematode Caenorhabditis elegans. Artemisia annua significantly increases survival under oxidative and heat stresses, however has no effects in response to ultraviolet stress. Then, we measured the in vivo changes in expression of stress-responsive genes by Artemisia annua using green fluorescence protein. The expression of hsp-16.2, known to be involved in response to heat stress, is significantly increased by Artemisia annua supplementation. An anti-oxidant gene, sod-3, was also up-regulated by Artemisia annua. However, both mean and maximum lifespan of Caenorhabditis elegans was not altered by dietary supplementation of Artemisia annua. These findings indicate that Artemisia annua confers health-promoting effects through increasing the resistance to environmental stressors and has no effect on lifespan in C. elegans. Our study suggests that Artemisia annua can be used for the development of novel natural therapeutics for diseases caused by environmental stressors.

Alzheimer’s disease is an age-related neurodegenerative disease. Extracellular aggregation of amyloid beta is a well-known hallmark of Alzheimer’s disease and believed to be critical for the development of Alzheimer’s disease. Lots of efforts have been put on finding an intervention alleviating amyloid beta toxicity. Electrolyzed-reduced water is generated near cathode by electrolysis of water and shown to have strong anti-oxidant and lifespan-extending activities in C. elegans. Here, we examined the effect of electrolyzed-reduced water on amyloid beta toxicity and investigated underlying cellular mechanism involved. Worms grown in media prepared with electrolyzedreduced water showed delayed amyloid beta-induced paralysis and age-related decline in motility. DAF-16, a FOXO transcription factor modulating insulin/IGF-1-like signal, is required for the protection against amyloid beta toxicity. These findings indicate that electrolyzed-reduced water can prevent amyloid beta toxicity via DAF-16 and suggest that electrolyzedreduced water could be utilized for the development of therapeutic intervention to Alzheimer’s disease.

Selenocysteine, a sulfur‑containing amino acid, can modulate cellular oxidative stress defense systems by incorporating into anti‑oxidant enzymes such as glutathione peroxidase and thioredoxin reductase. Selenocysteine can also prevent cancer, neurodegenerative diseases and cardiovascular diseases. A recent study revealed that dietary supplementation with selenocysteine can increase the resistance of Caenorhabditis elegans to environmental stressors and its lifespan. The objective of the present study was to identify the underlying mechanism involved in the lifespan‑extending effect of selenocysteine and the effect of selenocysteine on age‑associated pathophysiological changes. Lifespan assays with known long‑lived mutants of age‑1 (the ortholog of the phosphoinositide 3-kinase), clk‑1 (the ortholog of demethoxyubiquinone hydroxylase) and eat‑2 (a ligand-gated ion channel subunit) revealed that the effect of selenocysteine on lifespan specifically overlapped with that of the eat‑2 mutation, a genetic model of dietary restriction (DR). Selenocysteine mimicked the effect of DR on the bacterial dilution method. It required SKN-1 (the ortholog of mammalian nuclear factor-erythroid-related factor) for lifespan extension. In addition, selenocysteine significantly delayed the paralysis induced by human amyloid‑β gene, positively correlated with the incidence of Alzheimer's disease. The effect of selenocysteine on amyloid‑β‑induced toxicity was dependent on the nuclear localization of DAF‑16. Reduced survival caused by high‑glucose‑diet was recovered by selenocysteine. Selenocysteine also reduced the cellular level of reactive oxygen species known to be increased by high‑glucose‑diet. The results of the present study suggested that selenocysteine can mimic the effect of DR on lifespan and age‑associated pathophysiological alterations, providing scientific evidence for the development of DR mimetics using selenocysteine.

Recently, there have been reports that chronic insomnia acts as an insult in the brain, causing memory loss through the production of ROS, inflammation, and, Alzheimer's disease if persistent. Insomnia remains the leading cause of sleep disturbance and as such has serious implications for public health. Patients with Alzheimer's disease are also known to suffer from severe sleep disturbance. Meanwhile, vitexin is a key ingredient in Passiflora incarnata L (passion flower, PF) extract, which is known to help with sleep. This medicinal plant has been used as a folk remedy for sedation, anxiety and sleep since centuries ago, but the standardization work has not been done and the extent of the effect has not been clearly demonstrated. For this reason, we tried to test the possibility that repeated administration of PF could improve the memory by promoting hippocampal neurogenesis at the DBA/2 mice known have inherited sleep disorders, as well as preventive effects of Alzheimer's disease. Here, we found that vitexin, which is the main bioactive component of ethanol extracts from leaves and fruits (ratio; 8:2) of PF, confirmed the improvement of neurogenesis (DCX) of DBA/2 mice repeated PF oral administration by immunohistochemistry (IHC) and western blot analysis. PF-treated group showed increased the neurotrophic factor (BDNF) in the hippocampus compared with that of vehicle-treated group, but the inflammation markers Iba-1 (microglial marker) and COX-2 were inconsistent between the groups. However, we found COX-2 signal is essential for hippocampal neurogenesis according to the additional IHC experiments using COX-2 inhibitor and pIkappaB have shown. In addition, although prescription sleeping pills have been reported to show significant changes in appetite and metabolic rate from time to time, no changes in the feeding behavior, body weight, metabolic rate and body composition of the animals were observed by administration of PF. Interestingly, we found that short-term oral administration of PF displayed improved memory according to the water maze test. Quantitative analysis of Tau protein, which is a marker of Alzheimer's disease, was performed in the SD rats and DBA/2 mice by repeated PF oral administration and pTau/Tau values were significantly decreased in PF-treated group than vehicle-treated group. In conclusion, our results suggest that PF lead high hippocampal neurogenesis in the animals even in inherited sleep-disturbed animals. The increased hippocampal neurogenesis functionally enhanced memory and learning functions by repeated PF oral administration. These results identify PF as a potential therapy for enhancing memory functions and prevention of Alzheimer's disease through actions on the hippocampus.

The vine stem of Spatholobus suberectus Dunn (SS) is used as a traditional herbal medicine in China. Chinese herbal medicines are well known as natural bioactive compounds that can be used as new medicines, and their antioxidant and anticancer effects have also been reported. This study aimed to examine the anticancer effect of a high-pressure hot-water SS extract on rat C6 glioma cells. The SS extract effectively suppressed the viability and proliferation of C6 glioma cells through an antioxidant effect. Reactive oxygen species (ROS) levels in cancer cells are higher than that in normal cells. If the ROS level falls below that required for the growth of cancer cells, their rapid proliferation and growth can be suppressed. We also measured the induction of mitochondrial membrane depolarization and cell cycle arrest effect caused by the SS extract in C6 glioma cells through a FACS analysis. In addition, we observed an increase in STAT3, p53, E2F1, and p21 mRNA expression and a decrease in Bcl-2 mRNA expression by quantitative PCR. An increase in p21 protein expression of over 83% was observed through western blot analysis. All these data support the fact that the high-pressure hot-water SS extract has the potential to be used for glioma treatment.

OBJECTIVE: This study was performed to determine the effect of N-acetyl-L-cysteine, a modified sulfur-containing amino acid that acts as a strong cellular antioxidant, on the response to environmental stressors and on aging in C. elegans. METHOD: The survival of worms under oxidative stress conditions induced by paraquat was evaluated with and without in vivo N-acetyl-L-cysteine treatment. The effect of N-acetyl-L-cysteine on the response to other environmental stressors, including heat stress and ultraviolet irradiation (UV), was also monitored. To investigate the effect on aging, we examined changes in lifespan, fertility, and expression of age-related biomarkers in C. elegans after N-acetyl-L-cysteine treatment. RESULTS: Dietary N-acetyl-L-cysteine supplementation significantly increased resistance to oxidative stress, heat stress, and UV irradiation in C. elegans. In addition, N-acetyl-L-cysteine supplementation significantly extended both the mean and maximum lifespan of C. elegans. The mean lifespan was extended by up to 30.5% with 5 mM N-acetyl-L-cysteine treatment, and the maximum lifespan was increased by 8 days. N-acetyl-L-cysteine supplementation also increased the total number of progeny produced and extended the gravid period of C. elegans. The green fluorescent protein reporter assay revealed that expression of the stress-responsive genes, sod-3 and hsp-16.2, increased significantly following N-acetyl-L-cysteine treatment. CONCLUSION: N-acetyl-L-cysteine supplementation confers a longevity phenotype in C. elegans, possibly through increased resistance to environmental stressors.

Coelomocytes are specialized cells that continually and nonspecifically scavenge fluid from the body cavity through endocytosis in Caenorhabditis elegans. Our previous study revealed that coelmocytes were specifically required for dietary-restriction-induced longevity in C. elegans. In the present study, we examined the effect of coelomocyte ablation on the response to environmental stressors and reproduction in C. elegans. Coelomocytes were ablated using diphtheria toxin specifically expressed in coelomocytes. After exposing worms to 20 J/㎠/min of ultraviolet irradiation in vivo, the survival of the worms was monitored daily. To examine their response to heat stress, their survival after 10 h of 35° C heat shock was measured. Oxidative stress was induced using paraquat, and the susceptibility to oxidative stress was compared between wild-type control and coelomocyte-ablated worms. The total number of progeny produced was counted, and the time-course distribution of the progeny was determined. The worms with ablated coelomocytes showed reduced resistance to ultraviolet irradiation, but the ablation of coelomocytes had no effect on their response to heat or oxidative stress. The number of progeny produced during the gravid period was significantly decreased in the coelomocyte-ablated worms. These findings suggest that coelomocytes specifically modulate the response to ultraviolet irradiation and are required for normal reproduction in C. elegans. The findings could contribute to understanding of the mechanisms underlying dietary-restriction-induced longevity.

OBJECTIVE: The free radical theory of aging suggests that cellular oxidative damage caused by free radicals is a leading cause of aging. In the present study, we examined the effects of a well-known anti-oxidant amino acid derivative, selenocysteine, in response to environmental stress and aging using Caenorhabditis elegans as a model system. METHOD: The response to oxidative stress induced by H2O2 or ultraviolet irradiation was compared between the untreated control and selenocysteine-treated groups. The effect of selenocysteine on lifespan and fertility was then determined. To examine the effect of selenocysteine on muscle aging, we monitored the change in motility with aging in both the untreated control and selenocysteine-treated groups. RESULTS: Dietary supplementation with selenocysteine significantly increased resistance to oxidative stress. Survival after ultraviolet irradiation was also increased by supplementation with selenocysteine. Treatment with selenocysteine confers a longevity phenotype without an accompanying reduction in fertility, which is frequently observed in lifespan-extending interventions as a trade-off in C. elegans. In addition, the age-related decline in motility was significantly delayed by supplementation of selenocysteine. CONCLUSION: These findings suggest that dietary supplementation of selenocysteine can modulate response to stressors and lead to lifespan extension, thus supporting the free radical theory of aging.

Previous studies show that nutritional interventions with anti-oxidants have various health-promoting effects in several model organisms. Here, we examine the effects of S-allyl cysteine on resistance to environmental stressors and age-related physiological changes using C. elegans as a model system. S-allyl cysteine is a modified amino acid found in aged garlic extracts and known to have strong antioxidant activity. The survival of worms under oxidative-stress conditions significantly increased with supplementation of S-allyl cysteine. In addition, pre-treatment of S-allyl cysteine significantly increased resistance to both heat stress and ultraviolet irradiation. However, lifespan was not affected by S-allyl cysteine treatment. We also examined the effect of S-allyl cysteine on motility of C. elegans and found that S-allyl cysteine can retard the age-related decline of muscle tissue locomotive activity. S-allyl cysteine also significantly suppressed amyloid β-induced paralysis in Alzheimer's disease model animals. Taken together, our study indicates that dietary supplementation of S-allyl cysteine can improve health span and suggests that S-allyl cysteine can be used to develop novel health-promoting pharmaceuticals. Estudos anteriores mostram que intervenções nutricionais com antioxidantes têm vários efeitos promotores da saúde em vários organismos-modelo. Aqui, examinamos os efeitos da S-alil cisteína sobre a resistência a estressores ambientais e alterações fisiológicas relacionadas com a idade usando C. elegans como um sistema modelo. Salil cisteína é um aminoácido modificado encontrado em extratos de alho envelhecido e conhecido por ter forte atividade antioxidante. A sobrevivência de vermes sob condições de estresse oxidativo aumentou significativamente com a suplementação de S-alil cisteína. Além disso, o pré-tratamento com S-alil cisteína aumentou significativamente a resistência tanto ao estresse térmico como à irradiação ultravioleta. No entanto, o tempo de vida não foi afetado pelo tratamento com S-alil cisteína. Nós também examinamos o efeito da S-alil cisteína na motilidade de C. elegans e descobrimos que a S-alil cisteína pode retardar o declínio relacionado à idade da atividade locomotora do tecido muscular. A S-alil cisteína também suprimiu significativamente a paralisia induzida por amilóide β em animais-modelo da doença de Alzheimer. Tomados em conjunto, o nosso estudo indica que a suplementação dietética de S-alil cisteína pode melhorar a duração da saúde e sugere que S-alil cisteína pode ser usada para desenvolver novos produtos farmacêuticos de promoção da saúde.

Acanthopanax sessiliflorus, a plant mainly found in Korea, Japan, and China, has been used as a traditional medicine due to the bioactivities of some plant compounds. The antioxidant activity of A. sessiliflorus root in vivo and the anti-aging effect using Caenorhabditis elegans were studied. Pre-treatment with an A. sessiliflorus root extract significantly (p

A lab-scale sequential sludge digestion process which consists of a mesophilic anaerobic digester (MAD) and a thermophilic aerobic digester (TAD) was developed. Thermal, thermal–alkaline and long-term alkaline pre-treatments were applied to the feed sludge to examine their effects on sludge removal and methane production. Especially after thermal–alkaline pre-treatment, high COD removal was maintained; methane production rate was also drastically increased by improving the hydrolysis step of sludge degradation. Polymerase chain reaction–denaturing gel gradient electrophoresis indicated that bacterial communities were represented by three phyla (Firmicutes, Proteobacteria, Actinobacteria) and that Clostridium straminisolvens was the major bacterial species in MAD. Quantitative real-time PCR results indicated that Methanosaeta concilli was the major archaeal species in MAD, and that Ureibacillus sp. was the most abundant bacterial species in TAD.

Metagenome analysis was used to monitor changes in microbial population during the industrial-scale batch fermentation period (0, 15 days and 2 years). Genomic DNA was extracted from Bachu-Kimchi samples and was sequenced using GS Junior Titanium system, which yielded a total 6886, 6271, and 6621 reads from 0, 15 days and 2 years samples, respectively. Phylogenetic analysis based on 16S rRNA sequences showed clearly that microbial population was changed depending on the fermentation process (initial, rancid, and over-ripening stage). Wessella sp. and Leuconostoc sp. became the predominant group in microbial community at the optimum-rancid stage (15 days), but as the fermentation progressed more, the abundance of Lactobacillus sp. and Bacillus sp. increased (2 years). Moreover, the specific kimchi microbes in the industrial-scale fermentation process were isolated: Leuconostoc mesenteroides, Lactobacillus sakei, Lactobacillus plantarum, and Weissella koreensis.

To determine whether blood lead concentration is elevated in iron-deficient infants, blood lead and serum ferritin concentrations, serum iron/transferring iron-binding capacity (Fe/TIBC) and complete blood counts were measured in 30 iron deficient and 35 control infants, aged 6-24 months. All 30 iron-deficient infants received iron supplementation (ferric hydroxide-polymaltose complex, 6mg/kg Fe(3+)/day) for 1-6 months. Blood lead concentrations were measured in 18 of the iron deficient infants after their ferritin levels returned to the normal range. The geometric mean blood lead concentration was higher in iron deficient than in control infants (1.846 vs. 1.416μg/dL). After iron therapy, the blood lead levels of iron-deficient infants decreased significantly compared with pre-treatment levels (1.785 vs. 2.386μg/dL), and the hemoglobin and ferritin concentrations increased significantly. These findings indicate that iron deficiency increases blood lead concentrations in infants with very low blood lead concentrations.

Fpr3 and Fpr4 of Saccharomyces cerevisiae are nuclear FK506-binding proteins each containing an extended acidic domain in addition to the conserved FK506-binding/peptidylprolyl isomerase (PPIase) domain. Previous studies have shown that the PPIase domain regulates histone H3 methylation, while the acidic domain facilitates histone deposition and may regulate rDNA silencing. To gain insight into the role of FKBPs in maintaining chromatin structure, we examined the transcriptional profiles of fpr3 (-) and fpr4 (-) cells. Our results show that both proteins modulate the expression of a large number of genes randomly distributed throughout the genome, a pattern resembling those observed with yeast cells lacking other histone chaperones such as CAF1 and Asf1. Significant overlaps are found between nuclear FKBPs-modulated and the Asf1/CAF1-modulated genes. Thus, nuclear FKBPs appear to impact chromatin structure like other histone chaperones. Our analyses show that depleting Fpr4 causes no detectable chromatin structural change at the rDNA locus nor de-repression of transcription silencing at the locus, in contrast to a previous report. Furthermore, we demonstrate PPIase domain of the proteins represses transcription when tethered to the promoter of a reporter gene, suggesting that the PPIase domains can act on non-histone chromatin components, when brought to close proximity. The results thus provide a further demonstration for the elusive role of Fpr3 and Fpr4 in histone chaperones.

An effective two-stage sewage sludge digestion process, consisting of thermophilic aerobic digestion (TAD) followed by mesophilic anaerobic digestion (MAD), was developed for efficient sludge reduction and methane production. Using TAD as a biological pretreatment, the total volatile suspended solid reduction (VSSR) and methane production rate (MPR) in the MAD reactor were significantly improved. According to denaturing gradient gel electrophoresis (DGGE) analysis, the results indicated that the dominant bacteria species such as Ureibacillus thermophiles and Bacterium thermus in TAD were major routes for enhancing soluble organic matter. TAD pretreatment using a relatively short SRT of 1 day showed highly increased soluble organic products and positively affected an increment of bacteria populations which performed interrelated microbial metabolisms with methanogenic species in the MAD; consequently, a quantitative real-time PCR indicated greatly increased Methanosarcinales (acetate-utilizing methanogens) in the MAD, resulting in enhanced methane production.

The nematode Caenorhabditis elegans continues to serve as a useful model of life extension caused by dietary restriction. Using this model, downstream effectors of dietary restriction-induced longevity have been elucidated, including neuropeptides and cell-surface receptors. Although it remains possible that different forms of dietary restriction may utilize both specific and overlapping mechanisms to promote longevity, the nematode model has revealed roles for autophagy, metabolic energy-sensing and the hypoxic response. The nematode has also been used to identify specific tissues required for life extension via DR, including coelomocytes, intestine, and neurons.

Buckwheat (Fagopyrum esculentum) has been known for having strong anti-oxidant, anti-mutagenic, and anti-carcinogenic activities. The free radical theory of aging, also known as the oxidative stress theory of aging, claims that cellular oxidative damage accumulated with time is a major causal factor of aging. In the present study, we investigated the effect of buckwheat extracts on resistance to oxidative stress and aging using Caenorhabditis elegans as a model system. Survival under an oxidative-stress condition induced by paraquat increased markedly following 500mg/L buckwheat extracts treatment, suggesting lower cellular oxidative damage by buckwheat extracts. A lifespan assay also revealed that treatment of buckwheat extracts significantly extended both the mean and maximum lifespan in C. elegans. Interestingly, this lifespan-extension by buckwheat extracts was not accompanied by reduced fertility. These findings suggest that buckwheat extracts can confer longevity phenotype to C. elegans through its strong anti-oxidant activity and support the aging theory which emphasizes a pivotal role of oxidative stress during aging.

Background/Aims: Transcriptional repression of tumor suppressor genes is determined by the quantity of promoter hypermethylation. We analyzed the methylation quantity of CDKN2B in pediatric myelodysplastic syndromes (MDS). Methods: Quantitative measurement of CDKN2B methylation was performed in 25 pediatric MDS patients and 12 controls using pyrosequencing, and the result was compared with those from 74 adult MDS cases and 31 adult controls. The association between CDKN2B methylation quantity and factors related to prognosis including bone marrow blast percentage and karyotype was analyzed. Results: Pediatric MDS patients showed a higher methylation level (MtL) of CDKN2B than pediatric controls (2.94 vs. 1.62; p = 0.031) but a lower level than adult MDS patients (8.76; p < 0.001). MtL was higher in pediatric MDS cases with >5% blasts than in pediatric controls (3.78 vs. 1.62; p = 0.052). Pediatric MDS cases with abnormal karyotype showed a higher MtL than pediatric controls (5.95 vs. 1.62; p = 0.045). Conclusions: We confirmed that methylation of CDKN2B is associated with the pathogenesis and prognosis in pediatric MDS. The difference in MtLs between pediatric and adult MDS might be related to the physiological hypermethylation of tumor suppressor genes in aging.

Electrolyzed-reduced water (ERW) scavenges reactive oxygen species and is a powerful anti-oxidant. A positive correlation between oxidative stress and aging has been proved in many model organisms. In Caenorhabditis elegans, many long-lived mutants showed reduced fertility as a trade off against longevity phenotype. We aimed to study the effect of ERW on oxidative stress, fertility and lifespan of C. elegans. We also investigated the genetic pathway involved in the effect of ERW on resistance to oxidative stress and lifespan. We compared lifespan and fertility of worms in media prepared with distilled water and ERW. ERW significantly extended lifespan and increased the number of progeny produced. Then the effect of ERW on resistance to oxidative stress and lifespan of long-lived mutants was determined. ERW increased resistance to oxidative stress and lifespan of eat-2, a genetic model of dietary restriction, but had no effect on those of age-1, which is involved in insulin/insulin-like growth factor (IGF)-1-like signal. In addition, knockdown of daf-16, the downstream mediator of insulin/IGF-1-like signal, completely prevented the effect of ERW on lifespan. These findings suggest that ERW can extend lifespan without accompanying reduced fertility and modulate resistance to oxidative stress and lifespan via insulin/IGF-1-like signal in C. elegans.

Skin-derived precursor cells (SKPs) are multipotent, sphere-forming and embryonic neural crest‐related precu- rsor cells that can be isolated from dermis. It is known that the properties of porcine SKPs can be enhanced by leuke- mia inhibitory factor (LIF) which is an essential factor for the generation of embryonic stem cells in mice. In our pre- sent study, to enhance or maintain the properties of murine SKPs, LIF was added to the culture medium. SKPs were treated with 1,000 IU LIF for 72 hours after passage 3. Quantitative real time RT‐PCR was then performed to quantify the expression of the pluripotent stem cell specific genes Oct4, Nanog, Klf4 and c‐Myc, and the neural crest specific genes Snai2 and Ngfr. The results show that the expression of Oct4 is increased in murine SKPs by LIF treatment whereas the level of Ngfr is decreased under these conditions. Interestingly, LIF treatment reduced Nanog exp- ression which is also important for cell proliferation in adult stem cells and for osteogenic induction in mesenchymal stem cells. These findings implicate LIF in the maintenance of stem- ness in SKPs through the suppression of lineage differen- tiation and in part through the control of cell proliferation.

Tenebrio molitor are large insects and their larvae are consumed as food in many countries. The nutritional composition of T. molitor has been studied and contains high amounts of proteins, unsaturated fatty acids, and valuable minerals. However, the bioactivity of T. molitor has not been fully understood. We examined the effects of T. molitor extracts on resistance to oxidative stress and organism's lifespan using Caenorhabditis elegans as a model system. The response to heat shock and ultraviolet (UV) irradiation was monitored in vivo. The extracts from T. molitor showed significant effects on resistance to oxidative stress and UV irradiation and extend both mean and maximum lifespan of C. elegans. The number of progeny produced significantly increased in animals supplemented with T. molitor extracts. In addition, the expression of hsp-16.2 and sod-3 was markedly upregulated by supplementation with T. molitor extracts. These findings suggest that T. molitor extracts can increase response to stressors and extend lifespan by the induction of longevity assurance genes in C. elegans.

Electrolysis of water produces reduced water at the cathode and oxidized water at the anode. Electrolyzed-reduced water (ERW) has an extremely negative oxidation-reduction potential. ERW scavenges cellular reactive oxygen species (ROS) and suppresses single-strand breaks of plasmid DNA in bacteria. Here, we examined the effect of ERW on resistance to oxidative stress both in vitro and in vivo. Oxidative DNA damage in human lymphocytes was significantly alleviated by ERW by reducing cellular ROS levels. Caenorhabditis elegans grown in media prepared with ERW had increased resistance to oxidative stress caused by paraquat. We observed a significant effect of ERW on response to other stressors, including heat shock and UV-irradiation in C. elegans. These data indicate that the powerful anti-oxidant activity of ERW is due to its radical-scavenging activity and show, for the first time, that ERW could increase thermotolerance and resistance to UV-irradiation. These results suggest that ERW aids resistance to various environmental stresses.

Oxidative stress is generated by reactive oxygen species (ROS) including hydrogen peroxide (H(2)O(2)), hydroxyl radical (•OH ) and superoxide anion (O(2--)), which are produced as by-products of cellular metabolism. An imbalance in cellular redox status is a potent pathogenic factor that contributes to various chronic inflammatory diseases. In this study, we demonstrate that H(2)O(2 )decreases surfactant protein A, B and ABCA3 mRNA level, and increases SP-D mRNA level in human pulmonary lung epithelial cells. The decreased mRNA level of SP-A and SP-B were significant with a maximum inhibition of 79 and 87%, respectively by 150 µM H(2)O(2 )after 24 hrs of incubation. In addition, ABCA3 mRNA level was decreased with a maximum inhibition of 55% by 150 µM H(2)O(2 )after 12 hrs of incubation. In contrast, 150 µM H(2)O(2 )caused the SP-D mRNA level to increase to 200% of control after 8 hrs of incubation. The H(2)O(2)-induced gene repression or activation of SP-A, SP-B, SP-D and ABCA3 was blocked by pretreatment with the antioxidants N-acetyl-L-cysteine (NAC) and catalase. Furthermore, the inhibition of SP-A and SP-B was associated with reduced thyroid transcription factor -1 (TTF-1) DNA binding activity, and this reduced TTF-1 binding activity may be due to decreased TTF-1 protein expression level. The analyses of signal transduction pathways that may play a role in the regulation of gene expression by H(2)O(2 )using several specific inhibitors showed that U0126, an inhibitor of ERK1/2 upstream kinase MEK1/2, blocked both H(2)O(2)-induced inhibition of SP-A and SP-B gene expression, whereas SB203580, an inhibitor of p38 MAPK, partially blocked H(2)O(2)-mediated inhibition of SP-A gene expression but not SP-B expression. In contrast, AG-490, a specific inhibitor of JAK-STAT pathway, blocked H(2)O(2)-mediated inhibition of SP-B gene expression but not SP-A expression. Immunoblot analyses using specific phosphor-antibodies demonstrated that ERK1/2, p38 MAPK and STAT3 are phosphorylated by oxidative stress suggesting that H(2)O(2)-induced inhibition of SP-A and SP-B gene expression is associated with MAPK and JAKSTAT signaling pathway. These data, therefore, suggest that H(2)O(2 )affects SP-A and SP-B gene regulation by reducing TTF-1 DNA binding activity via MAPKs or STAT signaling pathways.

Aging is one of the most complicated biological processes in all species. A number of different model organisms from yeast to monkeys have been studied to understand the aging process. Until recently, many different age-related genes and age-regulating cellular pathways, such as insulin/IGF-1-like signal, mitochondrial dysfunction, Sir2 pathway, have been identified through classical genetic studies. Parallel to genetic approaches, genome-wide approaches have provided valuable insights for the understanding of molecular mechanisms occurring during aging. Gene expression profiling analysis can measure the transcriptional alteration of multiple genes in a genome simultaneously and is widely used to elucidate the mechanisms of complex biological pathways. Here, current global gene expression profiling studies on normal aging and age-related genetic/environmental interventions in widely-used model organisms are briefly reviewed.

MULTICELLULAR organisms must respond to changes in the environment by altering their pattern of allocation of resources between growth (soma) and reproduction (germ line) in order to maximize lifetime reproductive fitness. These responses are mediated by a complex set of still largely undefined interacting genetic networks that integrate energy acquisition and utilization, developmental and reproductive timing, and behavioral and sexual interactions. The tilt in the balance of these interactions toward early reproduction and the ultimate precedence of the germ line over the soma are hypothesized to be the ultimate causes of aging and senescence—bodies simply cannot be maintained indefinitely against inevitable environmental damage when that maintenance comes at the cost of a reproductive disadvantage. In its most extreme form, there will be no natural selection to maintain the soma once reproduction has ceased, unless there is some secondary contribution that the old can make to young relatives. Indeed, evolutionary theory predicts that one should see a “wave of death” late in life in most organisms (1–3). Thus, it is perhaps not surprising that mortality dramatically increases after female (hermaphrodite for Caenorhabditis elegans, which are essentially females that produce their own sperm; the hermaphrodite gonad is explained in detail in the Results) reproductive senescence in species that range from flies to humans (1,3–5). Caenorhabditis elegans has been an unexplained outlier in this trend, spending approximately 70% of its life in a reproductively senescent state—much more, for instance, than that of primates and other mammals (4). Is this simply an artifact of laboratory rearing or are there important portions of the biology of this nematode that have been overlooked in this important model of aging and senescence? Caenorhabditis elegans, which exist mostly as hermaphrodites, typically finish self-reproduction by about the fifth day of adult life (6,7). Surprisingly, these animals can live for another 2 weeks (6,8). Many mutations that extend organismal life span (Age) have been identified in the nematode. The first longevity mutant, age-1, shows as much as a 10-fold extension of somatic life (8,9); as in the wild type, all of this life extension is post-reproductive (10). In a few Age mutants, a modest extension of the period of self-fertility is seen (a relatively low number of progeny for a few days beyond wild type), but it comes at a cost of total progeny production (11–14). In addition to these results, two studies have recently demonstrated that C elegans can in fact display delayed reproductive senescence when hermaphrodite sperm is supplemented with sperm from males via mating (11,15); importantly, neither study determined the age limits of cross-fertility. It also now appears that there are multiple developmental checkpoints in which development and reproduction can be halted and then restarted in response to starvation (16–23). We would expect such factors to be important in longevity and reproduction because allocation to somatic maintenance versus reproductive output should vary in response to the availability and uncertainty of resources. Thus, there is reason to believe that standard picture of “normal” reproduction and longevity in C elegans, especially in the face of environmental mediators, may be incomplete. One gap in our understanding of reproductive life span is that no prior study has reported the functional life span of the germ line with respect to mating late in life. The latest age at which cross-fertility was examined in prior studies was Day 10 of adulthood (11). The maximum age at which hermaphrodites could become impregnated by males was not determined. Thus, the functional life span of the hermaphrodite germ line is not known when male mating is taken into account; life span (or functional life span) cannot be determined without observing function until cessation. Here, we measure germ line longevity by examining the capacity for C elegans hermaphrodites to respond to cross-fertilization after self-reproductive senescence—right up to the longevity barrier generated by somatic senescence. We find that a set of known longevity-extension mutants, representative of most classes of life-span extension, have little effect on germ line maintenance, whereas dietary input is critical. Thus, the structure of the genetic systems governing the interplay between reproduction and longevity in C elegans may be more complex than previously believed.

Gene fusion is a very rare mechanism that produces hemoglobin variants. Less than ten types of β-like hybrid globins have been reported to date. Herein we identified the first hybrid hemoglobin between Gγ- and β-globins in a five-year-old Korean male who had thalassemia minor feature and triplication of the HBA2 gene (αα/αααα). The novel globin originated from a 27,707-base pair deletion spanning from the HBG2 to HBB gene (NG_000007.3:g.42947_70653del). Its protein sequence included 13 N-terminal amino acids from Gγ-globin, five common amino acids from Gγ- and β-globins, and 128 amino acids from β-globin (Gγ through 13; β from 19). Molecular genetic analyses characterized the hybrid DNA and RNA. Mass spectrometry and de novo protein sequencing successfully identified the fusion peptide in the hybrid hemoglobin. We named this novel hybrid Hb Gγ-β Ulsan. The novel hemoglobin constituted 37.0% of the total hemoglobin and showed reduced oxygen affinity.

Purpose: The aim of this study was to evaluate the severity of disease in children with acute bronchiolitis according to the type of infected virus. Methods: From November 2007 to May 2015, 768 patients under 2 years of age who underwent real time-polymerase chain reac tion of nasopharyngeal aspirates admitted to the Department of Pediatrics of Dongguk University Ilsan Hospital for acute bronchi olitis were enrolled. Severe bronchiolitis was defined as presence of one or more kinds among tachypnea, chest retraction, needs of O2 inhalation or ventilator care. Results: The severity of bronchiolitis was increased with shorter fever duration (P

We used DNA microarrays to identify panels of transcriptional markers of aging that are differentially expressed in young (5-month) and old (25-month) mice of multiple inbred strains (129sv, BALB/c, CBA, DBA, B6, C3H, and B6C3F1). In the heart, age-related changes of five genes were studied throughout the mouse lifespan: complement component 4, chemokine ligand 14, component of Sp100-rs, phenylalanine hydroxylase, and src family associated phosphoprotein 2. A similar analysis in the brain (cerebellum) involved complement component 1q (alpha polypeptide), complement component 4, P lysozyme structural, glial fibrillary acidic protein, and cathepsin S. Caloric restriction inhibited age-related expression of these genes in both tissues. Parametric analysis of gene set enrichment (PAGE) identified several biological processes that are induced with aging in multiple mouse strains. We also tested the ability of dietary antioxidants to oppose these transcriptional markers of aging. Lycopene, resveratrol, acetyl-L-carnitine, and Tempol were as effective as caloric restriction in the heart, and α-lipoic acid and coenzyme Q10 were as effective as caloric restriction in the cerebellum. These findings suggest that transcriptional biomarkers of aging in mice can be used to estimate the efficacy of aging interventions on a tissue-specific basis.

Oxidative stress has been hypothesized to play a role in normal aging. The SKN-1 transcription factor regulates the response to oxidative stress and also is necessary for intestinal development in Caenorhabditis elegans. Using transcriptome analysis, we found that oxidative stress induces almost a thousand genes, including the antioxidant and heat-shock responses, as well as genes responsible for xenobiotic detoxification. There were also 392 down-regulated genes including many involved in metabolic homeostasis, organismal development, and reproduction. Many of these oxidative-stress-induced transcriptional changes are dependent on SKN-1 action; the induction of the heat-shock response is not. When we used RNAi to inhibit genes, we found that most had no effect on either resistance to oxidative stress or longevity; however two SKN-1-dependent genes, nlp-7 and cup-4, that were up-regulated by oxidative stress were found to be required for resistance to oxidative stress and for normal life span. nlp-7 encodes a neuropeptide-like protein, expressed in neurons, while cup-4 encodes a coelomocyte-specific, ligand-gated ion channel. RNAi of nlp-7 or cup-4 increased sensitivity to oxidative stress and reduced lifespan. Among down-regulated genes, only inhibition of ent-1, a nucleoside transporter, led to increased resistance to oxidative stress; inhibition had no effect on lifespan. In contrast, RNAi of nhx-2, a Na+/H+ exchanger, extended lifespan significantly without affecting sensitivity to oxidative stress. These findings show that oxidative stress causes a transcriptional shift from growth and maintenance towards the activation of cellular defense mechanisms; many of these transcriptional alterations are SKN-1-dependent.

Genomic instability and alterations in gene expression are hallmarks of eukaryotic aging. The yeast histone deacetylase Sir2 silences transcription and stabilizes repetitive DNA, but during aging or in response to a DNA break, the Sir complex relocalizes to sites of genomic instability, resulting in the desilencing of genes that cause sterility, a characteristic of yeast aging. Using embryonic stem cells, we show that mammalian Sir2, SIRT1, represses repetitive DNA and a functionally diverse set of genes across the mouse genome. In response to DNA damage, SIRT1 dissociates from these loci and relocalizes to DNA breaks to promote repair, resulting in transcriptional changes that parallel those in the aging mouse brain. Increased SIRT1 expression promotes survival in a mouse model of genomic instability and suppresses age-dependent transcriptional changes. Thus, DNA damage-induced redistribution of SIRT1 and other chromatin modifying proteins may be a conserved mechanism of aging in eukaryotes.

To investigate the global effects of vitamin E supplementation on aging, we used high-density oligonucleotide arrays to measure transcriptional alterations in the heart and brain (neocortex) of 30-month-old B6C3F1 mice supplemented with α- and γ-tocopherol since middle age (15 months). Gene expression profiles were obtained from 5- and 30-month-old controls and 30-month-old mice supplemented with α-tocopherol (1g/kg), or a mixture of α- and γ-tocopherol (500mg/kg of each tocopherol). In the heart, both tocopherol supplemented diets were effective in inhibiting the expression of genes previously associated with cardiomyocyte hypertrophy and increased innate immunity, while having a moderate effect on age-related transcriptional alterations linked to the stress response and protein synthesis. In the brain, induction of genes encoding ribosomal proteins and proteins involved in ATP biosynthesis was observed with aging and was markedly prevented by the mixture of α- and γ-tocopherol supplementation, but not by α-tocopherol alone. These results demonstrate that middle age-onset dietary supplementation with α- and γ-tocopherol can partially prevent age-associated transcriptional changes and that these effects are tissue- and tocopherol-specific.