Protective Effect of Liuwei Dihuangwan on Mitochondrial Damage in AD Model of Caenorhabditis Elegans / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of the extract of Liuwei Dihuangwan （LW） on mitochondrial damage in the Alzheimer's disease （AD） model of Caenorhabditis elegans （C. elegans）. MethodC. elegans transfected with human β-amyloid protein （Aβ） 1-42 gene was used as an AD model. The rats were divided into blank group， model group， metformin group （50 mmol·L-1）， and low， medium， and high dose （1.04， 2.08， 4.16 g·kg-1） LW groups. Behavioral methods were used to observe the sensitivity of 5-hydroxytryptamine （5-HT） in nematodes. Western blot was used to detect the expression of Aβ in nematodes. Total ATP content in nematodes was detected by the adenine nucleoside triphosphate （ATP） kit， and mitochondrial membrane potential was detected by the JC-1 method. In addition， the mRNA expression of Aβ expression gene （Amy-1）， superoxide dismutase-1 （SOD-1）， mitochondrial transcription factor A homologous gene-5 （HMG-5）， mitochondrial power-associated protein 1 （DRP1）， and mitochondrial mitoprotein 1 （FIS1） was detected by real-time fluorescence quantitative polymerase chain reaction （RT-PCR）. ResultThe extract of LW could reduce the hypersensitivity of the AD model of nematodes to exogenous 5-HT （P<0.05） and delay the AD-like pathological characteristics of hypersensitivity to exogenous 5-HT caused by toxicity from overexpression of Aβ in neurons of the AD model of nematodes. Compared with the blank group， in the model group， the mRNA expression of Aβ protein and Amy-1 increased （P<0.01）， and the mRNA expression of SOD-1 and HMG-5 decreased （P<0.01）. The mRNA expression of DRP1 and FIS1 increased （P<0.01）， and the level of mitochondrial membrane potential decreased （P<0.05）. The content of ATP decreased （P<0.01）. Compared with the model group， in the positive medicine group and medium and high dose LW groups， the mRNA expression of Aβ protein and Amy-1 decreased （P<0.05，P<0.01）， and the mRNA expression of SOD-1 and HMG-5 increased （P<0.01）. The mRNA expression of DRP1 decreased （P<0.05，P<0.01）， and that of FIS1 decreased （P<0.01）. The level of mitochondrial membrane potential increased （P<0.01）， and the content of ATP increased （P<0.05，P<0.01）. ConclusionThe extract of LW may enhance the antioxidant ability of mitochondria， protect mitochondrial DNA， reduce the fragmentation of mitochondrial division， repair the damaged mitochondria， adjust the mitochondrial membrane potential， restore the level of neuronal ATP， and reduce the neuronal damage caused by Aβ deposition.

Ezetimibe increases resistance to oxidative stress and extends lifespan mimicking dietary restriction in Caenorhabditis elegans

Ezetimibe is an approved drug for lowering plasma LDL (low-density lipoprotein) level via inhibition of cholesterol absorption. Derivatives of ezetimibe reduce inflammatory response and oxidative stress. In the present study, we investigated the effect of dietary supplementation with ezetimibe in response to environmental stressors and found that ezetimibe increases resistance to oxidative stress and ultraviolet irradiation. Ezetimibe also significantly extended lifespan accompanying reduced fertility, which is a common trade-off for longevity in C. elegans. Cellular level of reactive oxygen species was increased and the expression of stress-responsive genes, hsp-16.2 and sod-3, was induced by dietary supplementation with ezetimibe, suggesting a hormetic effect on oxidative stress response and lifespan. Ezetimibe also significantly prevented amyloid beta-induced toxicity and completely reversed increased mortality by high-glucose diet. Nuclear localization of DAF-16 required for the prevention of amyloid beta-induced toxicity was enhanced by ezetimibe supplementation. Lifespan assay using known long-lived mutants, age-1, clk-1, and eat-2, revealed that lifespan extension by ezetimibe specifically overlapped with that of eat-2 mutants, which are genetic models of dietary restriction. Effect of ezetimibe on lifespan of worms fed with diluted bacteria suggested that ezetimibe mimics the effect of dietary restriction on lifespan. These findings suggest that ezetimibe exhibits anti-oxidative and anti-aging effects through hormesis and works as a dietary-restriction mimetic on lifespan extension.

Anti-aging and mechanism of 2-O-cinnamic acid magnolol / 药学学报

Magnolol, a hydroquinone containing an allyl side chain, is one of the major active components of magnolia for antioxidation and anti-aging. To enhance the anti-aging activity and improve the intramolecular hydrogen bonding of magnolol, magnolol was reacted with cinnamic acid to obtain 2-O-cinnamic acid magnolol by esterification. The anti-aging activity of magnolol 2-O-cinnamate was investigated based on Caenorhabditis elegans model. The results showed that 2-O-cinnamic acid magnolol can reduce lipofuscin accumulation in the nematode body, and the effect is better than that of magnolol. 2-O-Cinnamic acid magnolol can extend nematode lifespan, reduce ROS levels in nematodes during normal aging and oxidative stress and improve nematode stress resistance under heat stress and oxidative stress. 2-O-Cinnamic acid magnolol could induce DAF-16 translocation from the cytoplasm to the nucleus and upregulate the expression of the sod-3 gene encoding superoxide dismutase in the nematode TJ356 expressing DAF-16 fused with GFP. 2-O-Cinnamic acid magnolol did not improve the survival rate of hsp-16.2 gene deficient nematodes under oxidative stress, indicating that 2-O-cinnamic acid magnolol improves stress resistance of nematodes under oxidative stress may be associated with sod-3 and hsp-16.2. Moreover, 2-O-cinnamic acid magnolol did not extend the lifespan of daf-16 and age-1 mutants, indicating that age-1 and daf-16 are required for 2-O-cinnamic acid magnolol to delay aging. It showed that magnolol 2-O-cinnamic acid has the potential to improve antioxidant capacity and delay aging, and the mechanism may be related to the insulin/insulin-like growth factor signaling pathway.

Caenorhabditis elegans deep lipidome profiling by using integrative mass spectrometry acquisitions reveals significantly altered lipid networks / 药物分析学报

Lipidomics coverage improvement is essential for functional lipid and pathway construction.A powerful approach to discovering organism lipidome is to combine various data acquisitions,such as full scan mass spectrometry(full MS),data-dependent acquisition(DDA),and data-independent acquisition(DIA).Caenorhabditis elegans(C.elegans)is a useful model for discovering toxic-induced metabolism,high-throughput drug screening,and a variety of human disease pathways.To determine the lipidome of C.elegans and investigate lipid disruption from the molecular level to the system biology level,we used integrative data acquisition.The methyl-tert-butyl ether method was used to extract L4 stage C.elegans after exposure to triclosan(TCS),perfluorooctanoic acid,and nanopolystyrene(nPS).Full MS,DDA,and DIA integrations were performed to comprehensively profile the C.elegans lipidome by Q-Exactive Plus MS.All annotated lipids were then analyzed using lipid ontology and pathway analysis.We annotated up to 940 lipids from 20 lipid classes involved in various functions and pathways.The biological in-vestigations revealed that when C.elegans were exposed to nPS,lipid droplets were disrupted,whereas plasma membrane-functionalized lipids were likely to be changed in the TCS treatment group.The nPS treatment caused a significant disruption in lipid storage.Triacylglycerol,glycerophospholipid,and ether class lipids were those primarily hindered by toxicants.Finally,toxicant exposure frequently involved numerous lipid-related pathways,including the phosphoinositide 3-kinase/protein kinase B pathway.In conclusion,an integrative data acquisition strategy was used to characterize the C elegans lipidome,providing valuable biological insights into hypothesis generation and validation.

Reproductive toxicity and associated mechanism of tricresyl phosphate on Caenorhabditis elegans / 环境与职业医学

Background Tricresyl phosphate (TCP) is mainly used as a flame retardant. Studies have confirmed that it has cytotoxicity and neurotoxicity, but its reproductive toxicity is not clear. Objective To investigate the reproductive toxicity and potential mechanism of TCP subacute exposure on Caenorhabditis elegans. Methods Caenorhabditis elegans were exposed to solvent control and 0.1, 1, 10, 100, and 1000 μg·L−1 TCP respectively for 72 h. Brood size and number of fertilized eggs in the uterus were detected to evaluate reproductive ability. The number of total germline cells and the relative area of gonad arm were measured to evaluate the development of gonads. The body length and body width of Caenorhabditis elegans were detected to evaluate growth and development. The activities of reactive oxygen species (ROS) and superoxide dismutase (SOD) in Caenorhabditis elegans, and the mitochondrial active oxygen metabolism genes (mev-1 and gas-1) of N2 nematodes were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) to evaluate oxidative stress. WS1433 transgenic nematodes and wild-type nematodes N2 were exposed to solvent control or TCP (0.1, 1, 10, 100, and 1000 μg·L−1) respectively. DNA damage in germ cells of WS1433 transgenic nematodes was detected, the relative expressions of DNA damage-related genes (hus-1, clk-2, cep-1, and egl-1) in N2 nematodes were detected by qRT-PCR to evaluate the effect of TCP exposure on genetic damage. Results Compared with the solvent control group (217.00 ± 12.20), the brood size of N2 nematodes in the 100 μg·L−1 and 1000 μg·L−1 TCP groups decreased (170.80 ± 11.51, 169.60 ± 10.52, P < 0.05). Compared with the solvent control group (18.43 ± 1.69), the number of fertilized eggs of N2 nematodes in the 100 μg·L−1 and 1000 μg·L−1 TCP groups decreased (13.47 ± 0.81, 11.95 ± 0.90, P < 0.05). Compared with the solvent control group (312.46 ± 77.4), the number of total germline cells of N2 nematodes in the 100 μg·L−1 and 1000 μg·L−1 TCP groups decreased (281.80 ± 12.98, 273.50 ± 8.53, P < 0.05). Compared with the solvent control group, the relative area of gonads of N2 nematodes in the 100 μg·L−1 and 1000 μg·L−1 TCP groups decreased by 13.83% and 17.25% respectively (P<0.05). Compared with the solvent control group [(1058.10±80.12) μm, (78.21±14.69) μm], the body length and body width of N2 nematodes in the 100 μg·L−1 and 1000 μg·L−1 TCP groups decreased (P<0.05). Compared with the solvent control group, the relative fluorescence intensity of ROS in nematodes in the 10, 100, and 1000 μg·L−1 TCP groups increased significantly (107.60%±1.02%, 105.90%±1.40%, and 106.40%±1.85%, respectively, P<0.05), and the activities of SOD were reduced (by 20.66%, 15.88%, and 16.44%, respectively, P<0.05). Compared with the solvent control group (1.3±1.3), the number of DNA-damaged germ cells of WS1433 nematodes in the 100 and 1000 μg·L−1 TCP groups increased significantly (2.4±0.3, 2.7±0.3, P<0.05); the expressions of mev-1 and gas-1 genes in N2 nematodes in the 10, 100 and 1000 μg·L−1 TCP groups decreased significantly (P<0.05); the expressions of hus-1 in the 0.1-1000 μg·L−1 TCP groups significantly increased (P<0.05); the expressions of clk-2 and egl-1 in the 100 and 1000 μg·L−1 TCP groups increased significantly (P<0.05); the expressions of cep-1 in the 1, 10, and 100 μg·L−1 TCP groups increased significantly (P<0.05). Conclusion TCP may cause reproductive damage to nematodes through oxidative stress and germ cell DNA damage.

Improvement effects and mechanism of thalidomide on Alzheimer ’s disease model of Caenorhabditis elegans / 中国药房

OBJECTIVE To study the improvement effects and mechan ism of thalidomide on Alzheimer ’s disease （AD）model of Caenorhabditis elegans . METHODS In this study ，the BR 5270 strain of C. elegans was used as AD model and BR 5271 strain as the control. The effects of thalidomide （0.5，2.0，6.0，15.0 mg/mL）on the motility of BR 5270 strains of C. elegans were studied by the basal slowing response assay ；the effects of thalidomide （0.5，2.0，6.0，15.0 mg/mL）on the survival time of BR 5270 strain of C. elegans were studied by life assay ；the effects of thalidomide （0.5，2.0，6.0 mg/mL）on learning and memory ability of BR 5270 strain of C. elegans were studied by short-term and long-term learning and memory assay. RT-PCR technology was used to study the effects of thalidomide （0.5，2.0，6.0 mg/mL）on mRNA expression of phosphatidylinositol 3-kinase（PI3K）/protein kinase B （Akt）signal pathway related genes （Age-1，Akt-1，Gsk-3）and calpain homologous gene （Clp-1）in BR 5270 strain of C. elegans . RESULTS After the intervention of thalidomide ，oscillation times of BR 5270 strain of C. elegans increased significantly within 30 s （except for 0.5 mg/mL group ），and the 10％ of maximum life span was prolonged significantly （only 0.5 mg/mL group ）；the short-term and long-term learning indexes were improved significantly （only 6.0 mg/mL group ）；mRNA expression of Age-1 and Akt-1（except for 0.5，2.0 mg/mL groups ）were increased significantly ，mRNA expression of Gsk-3（except for 0.5 mg/mL group ） and Clp-1 were decreased （P＜0.05 or P＜0.01）. CONCLUSIONS Thalidomide can ameliorate the dyskinesia of AD model of C. elegans，prolong the lifespan of this strain ，and enhance its learning and memory ability. Its mechanism of action may be related to activation of PI 3K/Akt signaling pathway and inhibition of calpain.

Caenorhabditis elegans como modelo de infección para el estudio de antimicrobianos / Caenorhabditis elegans as an infection model for the study of antimicrobials

Resumen A pesar de algunas limitaciones éticas, los animales juegan un papel importante como anfitriones sustitutos para investigar los mecanismos fisiopatológicos de enfermedades con el fin de indagar en ellos medicamentos contra diferentes patologías. Uno de los grandes problemas en salud pública a nivel mundial en el contexto farmacológico es la producción de antibióticos y la ocurrencia de resistencia microbiana, además, cada vez resulta más complejo el uso de modelos animales por las restricciones bioéticas actuales, no obstante, es necesario usar modelos simples en los estudios preliminares que permitan evaluar las interacciones huésped-patógeno-antimicrobiano. Al validar que Caenorhabditis elegans es susceptible a varias bacterias y además tiene la capacidad de responder a estímulos ambientales con cambios observables en el comportamiento tras ser alimentado con diversas bacterias, resulta muy útil usarlo en este tipo de investigaciones ya que tiene una vida promedio corta y no cuenta con restricciones éticas para su uso. Por lo anterior, en este artículo se revisa la susceptibilidad que tiene C.elegans de infectarse con diferentes bacterias, además, ya que aún no se ha validado completamente como modelo para poner a prueba antimicrobianos se propone que este nematodo es útil como modelo In vivo para evaluar infecciones y tratamientos antibacterianos.

Abstract Despite some ethical limitations, animals play an important role as surrogate hosts in investigating the pathophysiological mechanisms of disease in order to test drugs against different pathologies. One of the great problems in public health worldwide in the pharmacological context is the production of antibiotics and the occurrence of microbial resistance, the use of animal models is becoming increasingly complex due to current bioethical restrictions, however, it is necessary to use models simple in preliminary studies that allow evaluating host-pathogen-antimicrobial interactions. Validating that Caenorhabditis elegans is susceptible to various bacteria and also has the ability to respond to environmental stimuli with observable changes in behavior after being fed with various bacteria, it is very useful to use it in this type of research since it has a short average life and does not have ethical restrictions for its use. Therefore, this article reviews the susceptibility of C. elegance to become infected with different bacteria, in addition, since it has not yet been fully validated as a model to test antimicrobials, it is proposed that this nematode is useful as an in vivo model. to evaluate infections and antibacterial treatments.

Selective activity of diselenides against Aedes aegypti (Diptera: Culicidae) larvae

Abstract INTRODUCTION: Aedes aegypti (L.) is the major vector of arboviruses that causes serious public health concerns in tropical and subtropical countries. METHODS: We examined the larvicidal activity of 1,2-diphenyldiselenide [(PhSe)2] and 1,2-bis(4-chlorophenyl) diselenide [(p-ClPhSe)2] and determine its toxicity to different non-target organisms. RESULTS: (PhSe)2 and (p-ClPhSe)2 killed Ae. aegypti L3 larvae with LC50/24h values of 65.63 µM (20.48 mg/L) and 355.19 µM (135.33 mg/L), respectively. (PhSe)2 was not toxic to the four model organisms. CONCLUSIONS: (PhSe)2 is a larvicidal compound with selective action against Ae. aegypti larvae. The mechanisms of action of (PhSe)2 under field conditions remain to be investigated.

Antioxidant and antiaging effect of traditional Thai rejuvenation medicines in Caenorhabditis elegans / 中西医结合学报

OBJECTIVE@#This study explored the rejuvenation mechanisms of Thai polyherbal medicines using different approaches, including in vitro methods, as well as a well-defined nematode model, Caenorhabditis elegans.@*METHODS@#THP-R-SR012 decoction was selected from 23 polyherbal medicines, based on metal-chelating and chain-breaking antioxidant capacities. The influences of this extract on the survival and some stress biomarkers of C. elegans under paraquat-induced oxidative stress were evaluated. Furthermore, lifespan analysis and levels of lipofuscin accumulation were examined in senescent nematodes. The phytochemical profile of THP-R-SR012 was analyzed.@*RESULTS@#Supplementation with THP-R-SR012 decoction significantly increased the mean lifespan and reduced the oxidative damage to C. elegans under oxidative stress conditions. Further, THP-R-SR012 supplementation slightly influenced the lifespan and the level of lipofuscin accumulation during adulthood. Antioxidant-related phytochemical constituents of THP-R-SR012 decoction were rutin, naringenin, 3,4-dihydroxybenzoic acid, gallic acid, glycyrrhizic acid, demethoxycurcumin and 18α-glycyrrhetinic acid.@*CONCLUSION@#The antioxidant potential of THP-R-SR012 was due to its scavenging properties, its enhancement of antioxidant-related enzyme activities, and the presence of the antioxidant-related compound. These results support the traditional use of THP-R-SR012 decoction as a tonic for nourishing and strengthening the whole body.

Study on the Effects of Atractylodes macrocephala Ethanol Extract on Life Span of Caenorhabditis elegans and Its Mechanism / 中国药房

OBJECTIVE： To study the effects of Atractylodes macrocephala ethanol extract （AM） on life span of Caenorhabditis elegans（called N 2 nematode for short ），and to investigate its mechanism based on transcription factor SKN- 1/ nuclear factor E 2 related factor 2（Nrf2）. METHODS ：N2 nematode were divided into blank control group ，positive control group （100 μ mol/L curcumin，similarly hereinafter ），AM low-dose ，medium-dose and high-dose groups （100，200，300 μ g/mL， similarly hereinafter ）. The effects of AM on the life span （by average survival time ）of N 2 nematode under normal condition and oxidant stress condition （40 mmol/L H 2O2）as well as its effects on reproductive capability （by the number of filial generation ）of N2 nematode under normal condition were investigated . 700 μmol/L H2O2 was used to establish neuroblastoma cells N 2a oxidant stress model. Effects of positive control ，low-dose，medium-dose and high-dose of AM on the survival rate of model cells were detected by MTT method. After human embryonic renalepithelial cells 293T were transfected with Nrf 2-ARE plasmid ， the effects of positive control and AM on luciferase activity of Nrf2-ARE were detected by luciferase reporter gene method at low，medium and high dose for 24 h and at medium dose for 12，18 and 24 h. RT-PCR was used to detect the effects ofpositive control ，low-dose，medium-dose and high-dose of AM on the mRNA expression of downstream genes NQO- 1 and HO- 1 of Nrf 2 in N 2a cells as well as mRN A expression of en@hactcm.edu.cn downstream genes GCS- 1，GST-7，GST-10，HSP-60，HSP- 16.2 and SOD- 3 of SKN- 1 in N 2 nematode. RESULTS ：Compared with blank control group ，average survival time of N 2 nematode under normal and oxidant stress condition was significantly prolonged in positive control group and AM groups ；the number of filial generation on the first day （except for AM high-dose group ），the number of filial generation on the second day （except for AM low-dose group ） and the total number of filial generation （except for AM low-dose group ） were increased significantly（P＜0.05 or P＜0.01）. The survival rate of N 2a cells in positive control group ，AM medium-dose and high-dose groups were significantly higher than that of model group （P＜0.05 or P＜0.01）. Compared with blank control group ，Nrf2-ARE luciferase relative activity of 293T cells in positive control group and AM groups as well as Nrf 2-ARE luciferase relative activity of 293T cells in AM medium-dose group after different time of treatment were increased significantly （P＜0.01），in dose-dependent and time-dependent trend. Compared with blank control group ，mRNA relative expression of HO- 1 and NQO- 1（except for positive control group ），GCS-1（except for AM low-dose group ），GST-7（except for positive control group and AM low-dose group ）， GST-10 and HSP- 60（except for AM low-dose group ），HSP-16.2（except for positive control group and AM low-dose group ）and SOD-3 （except for positive control group and AM low-dose group ） were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS：AM can prolong the life span of N 2 nematode under normal and oxidant stress condition and improve the its reproductive capacity ，the mechanism of which may be associated with the activation of SKN- 1/Nrf2 signaling pathway.

Effects of VHL inhibitor on rotenone-induced Caenorhabditis elegans model of Parkinson′s disease / 中国药科大学学报

@#To investigate the effects of VHL (von Hippel-Lindau) inhibitor on Caenorhabditis elegans (C.elegans) model of Parkinson''s disease (PD),C.elegans were exposed to rotenone and treated with VHL inhibitor VH298.The death,dopaminergic neurodegeneration and mitochondrial unfolded protein response (mito-UPR) of transgenic strains with the markers zcIs9 and otIs181 exposed to different concentrations of rotenone were investigated. The death,dopaminergic neurodegeneration,and changes of behaviors including head thrashes,body bends and foraging behavior of C.elegans model of PD treated with different concentrations of VH298 were explored.The results showed that different concentrations of rotenone can lead to the death,dopaminergic neurodegeneration and abnormal mito-UPR of transgenic nematodes with zcIs9; otIs181,while the VHL inhibitor can decrease the death rate and alleviate dopaminergic neurodegeneration of rotenone-induced C.elegans model of PD.The VHL inhibitor can also attenuate the behavioral abnormalities of head thrashes,body bends and foraging behavior of C.elegans model.These results suggest that rotenone may cause mitochondrial damage in the transgenic nematodes with zcIs9; otIs181, and then destroy mitochondrial homeostasis,thereby resulting in dopaminergic neurodegeneration and death of the nematodes. The VHL inhibitor VH298 may promote the survival of rotenone-induced C.elegans model of PD,and alleviate dopaminergic neurodegeneration,thereby improving the behavioral abnormalities of C.elegans model of PD.

Efecto de disrupción endocrina de la genisteína sobre caenorhabditis elegans

La genisteína es una isoflavona presente en la soya, de alto consumo en la población infantil por su uso como sucedáneo de la leche materna, sin embargo, poco se conoce acerca de los efectos a nivel endocrino. En este trabajo, Caenorhabditis elegans se utilizó como modelo para evaluar el efecto de disrupción endocrina de la genisteína a través de letalidad, crecimiento, reproducción, almacenamiento de lípidos y cambios en la expresión de genes de respuesta al estrés (hsp-3, sod-4 y gpx-4). Los resultados indicaron que, aunque la genisteína no indujo letalidad, sí promovió la reproducción, el aumento de la longitud del cuerpo, el incremento en la expresión de genes relacionados con estrés celular y estrés oxidativo y la acumulación lipídica. En conclusión, la genisteína generó efectos relacionados con el efecto de disrupción endocrina en C. elegans, muy probablemente a través de mecanismos de estrés oxidativo.

Genistein is an isoflavone present in soy, which children highly consume as a substitute for breast milk; however, little is known about its effects at the endocrine level. This paper used Caenorhabditis elegans as a model to evaluate the endocrine disrupting effect of genistein through lethality, growth, reproduction, lipid storage, and changes in the expression of stress response genes (hsp-3, sod- 4, and gpx-4). The results indicated that, although genistein did not induce lethality, it did promote reproduction and increased body length, expression of genes related to cellular stress and oxidative stress, and lipid accumulation. In conclusion, genistein produced effects related to endocrine disruption on C. elegans, most likely through oxidative stress mechanisms.

Toxicity and metabolism of 3-bromopyruvate in Caenorhabditis elegans / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

In this study, we aimed to evaluate the toxic effects, changes in life span, and expression of various metabolism-related genes in Caenorhabditis elegans, using RNA interference (RNAi) and mutant strains, after 3-bromopyruvate (3-BrPA) treatment. C. elegans was treated with various concentrations of 3-BrPA on nematode growth medium (NGM) plates, and their survival was monitored every 24 h. The expression of genes related to metabolism was measured by the real-time fluorescent quantitative polymerase chain reaction (qPCR). Nematode survival in the presence of 3-BrPA was also studied after silencing three hexokinase (HK) genes. The average life span of C. elegans cultured on NGM with 3-BrPA was shortened to 5.7 d compared with 7.7 d in the control group. hxk-1, hxk-2, and hxk-3 were overexpressed after the treatment with 3-BrPA. After successfully interfering hxk-1, hxk-2, and hxk-3, the 50% lethal concentration (LC50) of all mutant nematodes decreased with 3-BrPA treatment for 24 h compared with that of the control. All the cyp35 genes tested were overexpressed, except cyp-35B3. The induction of cyp-35A1 expression was most obvious. The LC50 values of the mutant strains cyp-35A1, cyp-35A2, cyp-35A4, cyp-35B3, and cyp-35C1 were lower than that of the control. Thus, the toxicity of 3-BrPA is closely related to its effect on hexokinase metabolism in nematodes, and the cyp-35 family plays a key role in the metabolism of 3-BrPA.

Nutraceutical Potential and Antipsychotic Activity of Local Dry Fruit Waste – Juglans regia

Aim: To identify the biologically active components in shells of Juglans regia and study its nutraceutical potential and antipsychotic activity for effective waste management. Study Design: Biochemical and in vivo analyses of plant extract using established protocols. Place and Duration of Study: Sample extraction at Department of Food Science and technology, School of Biotechnology and Bioinformatics, DY Patil deemed to be University, Navi Mumbai, India; sample components identification at Sophisticated Analytical Instrument facility (SAIF), Indian Institute of Technology (IIT), Mumbai, India; and in vivo studies for antipsychotic activity using Caenorhabditis elegans at Department of Life Science and Biochemistry, St. Xavier’s College, Mumbai, India between November 2018 and May 2019. Methodology: The shells of Juglans regia were milled and the extract was prepared using Soxhlet extraction at 60oC using methanol as solvent. The GCMS analysis of the extract was carried out using a GC JEOL – The Accu TOF. Antipsychotic activity was studied using pharyngeal pumping assay in Caenorhabditis elegans. Results: GC-MS analysis of methanolic extract of shells of Juglans regia revealed the presence of Tridecanoic acid, Acetoxyacetic acid, nonyl ester, 2-hexenal, 2-ethyl, Eicosanoic acid, phenylmethyl ester, Undecane, Benzeneacetic acid decyl ester, (1-pentyl-allyoxymethoxy-methyl)-benzene), 9,12-octadecadienoic acid(Z,Z), phenylmethyl ester, Benzyl oxytridecanoic acid, 6,9,12- octadecatrienoic acid, phenylmethyl ester (Z,Z), 9- octadecanoic acid (Z), phenylmethyl ester, 9,12,15- octadecatrienoic acid, Z [(trimethyl (sil)oxy, 1 – trimethyl (sily)oxy] ethyl ester (Z,Z,Z). Furthermore, behavioural assay done using Caenorhabditis elegans as a model organism showed that the sample exerted antipsychotic activity at lowest concentration. Conclusion: The shells of Juglans regia being a natural source, can be used as an alternative to the synthetic antipsychotic drugs that have side effects. Our current work suggests that the walnut shells that end up into trash bins are an excellent source of effective natural biologically active compounds.

Differences in protein expression associated with ivermectin resistance in Caenorhabditis elegans / Diferenças na expressão de proteínas associadas à resistência à ivermectina em Caenorhabditis elegans

Abstract The indiscriminate administration of synthetic anthelmintics such as ivermectin contributes to the selection of subpopulations capable of resisting the drugs' effects. To understand the mechanisms of ivermectin resistance in Caenorhabditis elegans, this study attempted to identify molecular targets. C. elegans lineages that were sensitive and resistant to ivermectin were used. Collected nematodes were added to an extraction buffer and macerated in liquid nitrogen for protein extraction. The extracted proteins were separated according to molecular weight by SDS-PAGE to verify their integrity. Subsequently, proteins from both lineages were separated using two-dimensional electrophoresis. The gels were analyzed and the relevant spots were excised and identified by mass spectrometry (NanoESI-Q-TOF and MASCOT®) and subsequently assessed by GO enrichment and STRING® analyses. The increased expression of proteins associated with high metabolic activity, such as ATP-2 and ENOL-1, which are responsible for ATP synthesis, was observed. Furthermore, proteins with involvement in mediating muscular function (MLC-1, ACT-1, and PDI-2), signaling (FAR-1 and FAR-2), and embryo development (VHA-2) were identified. Protein interaction analysis indicated that the majority of the identified proteins in the resistant lineages participated in the same reaction triggered by ivermectin.

Resumo A administração indiscriminada de anti-helmínticos sintéticos, como a ivermectina, contribui para a seleção de subpopulações capazes de resistir ao efeito das drogas. Para entender os mecanismos de resistência à ivermectina em Caenorhabditis elegans, este estudo visou identificar alvos moleculares. Portanto, linhagens de C. elegans sensíveis e resistentes à ivermectina foram utilizadas. Os nematóides coletados foram adicionados ao tampão de extração e macerados em nitrogênio líquido para obtenção das proteínas. As proteínas extraídas foram separadas por peso molecular em SDS-PAGE para verificar sua integridade. Posteriormente, as proteínas de ambas as linhagens foram separadas por eletroforese bidimensional. Os géis foram analisados, os spots relevantes foram excisados e identificados por espectrometria de massa (NanoESI-Q-TOF e MASCOT®), em seguida, analisados ​​em seus termos de GO e STRING®. A expressão aumentada de proteínas associadas à alta atividade metabólica, como as proteínas ATP-2 e ENOL-1, responsáveis ​​pela síntese de ATP, foi observada. Além disso, foram identificadas as proteínas responsáveis ​​pelo controle da função muscular (MLC-1, ACT-1 e PDI-2), sinalização (FAR-1 e FAR-2) e desenvolvimento embrionário (VHA-2). A análise das interações proteicas indicou que a maioria das proteínas identificadas na cepa resistente participa da mesma reação desencadeada pela ivermectina.

Toxicity and oxidative damage of different silver nanoparticles to Caenorhabditis elegans / 中国药理学与毒理学杂志

Objective: To investigate the toxic effects of three types of silver nanoparticles (AgNPs): 20 and 50 nm uncoated nanosilver (AgNPs-20 and AgNPs-50) and 20 nm polyvinylpyrrolidone coated nanosilver (AgNPs-PVP-20) to Caenorhabditis elegans (C.elegans). Methods: Wild type C. elegans were exposed to 1% hydroxypropyl methylcellulose (vehicle group) and AgNPs-20, AgNPs-50, AgNPs-PVP-20 (1,10 and 100 mg·L-1) for 48 h, respectively. The survival, head thrash and body bend frequency of the nematodes were recorded. The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH) in nematodes were detected by DCFH-DA fluorescence probe, TBA method and colorimetry, respectively. Results: Compared with vehicle group, the survival rate of nematodes was decreased in each dose group of AgNPs-PVP-20,10 and 100 mg·L-1groups of AgNPs-50 and AgNPs-20 (P<0.01). The relative mean lifespan of nematodes in each dose group of AgNPs-PVP-20 was significantly decreased (P<0.01), while that of AgNPs-20, AgNPs-50 (10 and 100 mg·L-1) was decreased (P<0.05). The frequency of head thrashes in 10 and 100 mg·L-1of the three AgNPs groups was decreased (P<0.05), and the frequency of body bends in each dose group of AgNPs-PVP-20 was significantly reduced (P<0.01). The body bends in 100 mg·L-1groups of AgNPs-20 and AgNPs-50 were decreased (P<0.05). The fluorescence intensity of ROS in the three AgNPs groups was significantly higher than that of the control group (P<0.01). Compared with the control group, the content of MDA was increased and the content of GSH decreased (P<0.05) in AgNPs-PVP-20 (10 and 100 mg·L-1) groups. There was no significant difference in MDA or GSH contents between AgNPs-50 and control groups. Conclusion: The relative mean lifespan of nematodes, the head thrash frequency and body bend frequency of nematodes were decreased to different extents with the increasing dose in each of the AgNPs groups, whereas the oxidative stress level of nematodes was increased unexpectedly in all the test samples. The toxic effect of AgNPs-PVP-20 is the strongest, followed by AgNPs-20 and AgNPs-50.

Evaluation of pathogenicity of Acinetobacter venetum by using Caenorhabditis elegans / 预防医学

Objective@#To evaluate the pathogenicity of Acinetobacter venetum（Av），which is expected to be used as an environmental remediation agent，using Caenorhabditis elegans（C.elegans）. @*Methods@#The C.elegans were cultured on the media loaded with E.coli OP50 and Av，respectively. The pathogenicity of Av was evaluated by observing the effects of Av on the growth，movement，digestive function，lifespan and reproduction of C.elegans，compared with that of another evaluation system according to NY 1109-2017 General Biosafety Standard for Microbial Fertilizers.@*Results@#By C. elegans system，it was found that the body length，width，head thrash frequency，body bending frequency and average lifespan [（13.5±0.4）d vs.（13.7±0.4）d] of adult nematodes in the Av group were not significantly different from those in the OP50 group（all P>0.05）；while the average time of defecation cycle in the Av group shortened，the total number of progenies in the Av group increased by 18.7%（all P<0.05）. According to NY1109-2017 General Biosafety Standard for Microbial Fertilizers，it was found that the oral LD50 values for both male and female mice were more than 10 g/kgbw，which was practically non-toxic；the pathogenicity test of acute intraperitoneal injection showed that the animals did not have signs of poisoning，deaths or any abnormalities in gross anatomy；Av had no irritation to damaged skin and eyes of rabbits；the hemolysis test was negative；Av was sensitive to seven antibiotics and was medium to one antibiotic. @*Conclusion@#Av is not pathogenic. C. elegans can be used in early screening for the pathogenicity of environmental remediation agents.

Anti-oxidative Effect of Piperine from Piper nigrum L. in Caenorhabditis elegans

Piper nigrum L. (Piperaceae), which is a well-known food seasoning, has been used as a traditional medicine for the treatment of vomiting, abdominal pain, diarrhea and anorexia in Korea, China and Japan. Methanol extract from the fruit of P. nigrum was successively partitioned as n-hexane, methylene chloride, ethyl acetate, n-butanol and H₂O soluble fractions. Among those fractions the ethyl acetate soluble fraction showed the most potent DPPH radical scavenging activity, and piperine was isolated from the ethyl acetate fraction. To know the antioxidant activity of piperine, we tested the activities of superoxide dismutase (SOD) and catalase together with oxidative stress tolerance and intracellular ROS level in Caenorhabditis elegans. To investigate whether piperine-mediated increased stress tolerance was due to regulation of stress-response gene, we quantified SOD-3 expression using transgenic strain including CF1553. Consequently, piperine enhanced SOD and catalase activities of C. elegans, and reduced intracellular ROS accumulation in a dose–dependent manner. Moreover, piperine-treated CF1553 worms exhibited significantly higher SOD-3::GFP intensity.

Leucosceptroid B from glandular trichomes of Leucosceptrum canum reduces fat accumulation in Caenorhabditis elegans through suppressing unsaturated fatty acid biosynthesis / 中国天然药物

Obesity that is highly associated with numerous metabolic diseases has become a global health issue nowdays. Plant sesterterpenoids are an important group of natural products with great potential; thus, their bioactivities deserve extensive exploration. RNA-seq analysis indicated that leucosceptroid B, a sesterterpenoid previously discovered from the glandular trichomes of Leucosceptrum canum, significantly regulated the expression of 10 genes involved in lipid metabolism in Caenorhabditis elegans. Furthermore, leucosceptroid B was found to reduce fat storage, and downregulate the expression of two stearoyl-CoA desaturase (SCD) genes fat-6 and fat-7, and a fatty acid elongase gene elo-2 in wild-type C. elegans. In addition, leucosceptroid B significantly decreased fat accumulation in both fat-6 and fat-7 mutant worms but did not affect the fat storage of fat-6; fat-7 double mutant. These findings indicated that leucosceptroid B reduced fat storage depending on the downregulated expression of fat-6, fat-7 and elo-2 and thereby inhibiting the biosynthesis of the corresponding unsaturated fatty acid. These findings provide new insights into the development and utilization of plant sesterterpenoids as potential antilipemic agents.

The review of ionizing radiation model studies using Caenorhabditis elegans / 中华放射医学与防护杂志

With the increasing prominence of radiotherapy in the treatment of tumors,how to reduce the side effects of radiotherapy has become a hot research topic gradually.Caenorhabditis elegans (C.elegans) has become a good model for studying ionizing radiation due to its unique advantages.In recent years,there's increasing study on ionizing radiation by using nematode model.In this paper,the effects of ionizing radiation on nematode and underlying mechanisms are reviewed briefly.