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mgr-mir-9 implicates Meloidogyne graminicola infection in rice by targeting the effector MgPDI

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Journal of Integrative Agriculture 2023年第5期22卷 1445-1454页

作者： TIAN Zhong-ling ZHOU Jia-yan ZHENG Jing-wu HAN Shao-jie Key Laboratory of Pollution Exposure and Health Intervention of Zhejiang Province Interdisciplinary Research Academy(IRA)Zhejiang Shuren UniversityHangzhou 310015P.R.China Laboratory of Plant Nematology Institute of BiotechnologyCollege of Agriculture&BiotechnologyZhejiang UniversityHangzhou 310058P.R.China Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insect Pests Zhejiang UniversityHangzhou 310058P.R.China

MicroRNAs (miRNAs),a class of small non-coding RNAs,are crucial endogenous gene regulators in a range of animals,including plant-parasitic *** graminicola is an obligate sedentary endoparasite of rice and causes significant yield losses.A number of studies focused on the roles of *** effectors during the parasitic process;however,how nematode miRNAs regulate its effectors needs *** this research,we analyzed a cluster of *** miRNAs obtained at the second-stage juveniles (J2s) stage that are closely linked to the regulation of *** *** are 49 767 105 total clean reads obtained from three libraries.A total of 233 known miRNAs and 21 novel miRNAs were *** the known miRNAs,mgr-lin-4,mgr-mir-1,mgr-mir-100,mgrmir-86,mgr-mir-279,mgr-mir-87,mgr-mir-71,mgr-mir-9,mgr-mir-50,mgr-mir-72,and mgr-mir-34 are the most abundant11 miRNAs ***,the expression levels of selected miRNAs were validated by real-time quantitative *** hypothesized that these miRNAs might regulate the expression of secreted effectors during the J2s stage to facilitate its *** with this,we found that mgr-mir-9 targets MgPDI,an important *** effector *** addition to that,J2s treated with mgr-mir-9 mimics showed down-regulation of MgPDI expression and reduced reproductive ability,alluding mgr-mir-9 is involved in nematode *** results provide novel insight into the regulatory functions of *** miRNAs during the infection and identify miRNAs and their effector targets as potential key management targets to limit parasite survival during the early stages of infection.

关键词： microRNA function Meloidogyne graminicola deep sequencing Dual-Luciferase Reporter assay System proteindisulfideisomerase

Hepatitis C virus antigens enzyme immunoassay for one-step diagnosis of hepatitis C virus coinfection in human immunodeficiency virus infected individuals

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World Journal of Hepatology 2019年第5期11卷 442-449页

作者： Ke-Qin Hu Wei Cui Susan D Rouster Kenneth E Sherman Division of GI/Hepatology University of CaliforniaIrvineSchool of MedicineOrangeCA 92868United States Division of Digestive Diseases University of Cincinnati College of MedicineCincinnatiOH 45267United States

BACKGROUND Current diagnosis of hepatitis C virus(HCV)infection requires two sequential steps:testing for anti-HCV followed by HCV RNA PCR to confirm *** have developed a highly sensitive and specific HCV-antigens enzyme immunoassay(HCV-Ags EIA)for one-step diagnosis of viremic HCV *** To assess the clinical application of the HCV-Ags EIA in one-step diagnosis of viremic HCV infection in human immunodeficiency virus(HIV)-coinfected *** The study blindly tested HCV-Ags EIA for its performance in one-step diagnosing viremic HCV infection in 147 sera:10 without HCV or HIV infection;54 with viremic HCV monoinfection;38 with viremic HCV/HIV coinfection;and 45 with viremic HCV and non-viremic HIV *** Upon decoding,it was 100%accordance of HCV-Ags EIA to HCV infection status by HCV RNA PCR *** five sera with HCV infection,HCV RNA was as low as 50-59 IU/mL,and four out of five tested positive for HCV-Ags ***,it was also 100%accordance of HCV-Ags EIA to HCV infection status by HCV RNA PCR in 83 sera with HCV and HIV coinfection,regardless if HIV infection was active or *** The modified HCV-Ags EIA has a lower detection limit equivalent to serum HCV RNA levels of approximately 100 IU/*** is highly sensitive and specific in the setting of HIV coinfection,regardless of HIV infection status and CD4 *** data support the clinical application of the HCV-Ags EIA in one-step diagnosis of HCV infection in HIV-infected individuals.

关键词： Hepatitis C virus Hepatitis C virus antigens Hepatitis C virus core antigen Hepatitis C virus diagnostic test Diagnostic assay Enzyme immunoassay

Cytotoxicity evaluation and hepatoprotective potential of bioassay guided fractions from Feronia limmonia Linn leaf

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Asian Pacific Journal of Tropical Biomedicine 2011年第6期1卷 443-447页

作者： Mahendra Jain Rakhee Kapadia Ravirajsinh N Jadeja Menaka C Thounaojam Ranjitsinh V Devkar SH Mishra Herbal Drug Technology Laboratory Pharmacy DepartmentFaculty of Technology and EngineeringThe M.S.University of Baroda Novel Drug Delivery System Laboratory Pharmacy DepartmentFaculty of Technology and EngineeringThe M.S.University of Baroda Division of Phytothrapeutics and Metabolic Endocrinology Department of ZoologyThe M.S.University of Baroda

Objective:To evaluate the cytotoxicity and hepatoprotective potentials of extracts,fractions or isolated compound from the leaves of Feronia limonia(***).Methods:Qualitative phytochemical analysis of extracts,fractions or compound was performed by means of thin layer chromatography and spectroscopic ***%purity of compound was measured by analytical ***,fractions or compound have been individually evaluated for their cytotoxicity effects(10,20,100,250,500,750 and 1 000 μg/mL).Based on the inhibitory concentration(IC_(50)) obtained from the cell viability assay,graded concentrations of extracts,fractions or isolated compound were assessed(10,20,50,100,200 μg/mL) for its hepatoprotective potential against CCl_4-induced hepatotoxicity by monitoring activity levels of serum glutamatic pyruvatic transaminase(SGPT) and serum glutamic oxaloacetic transaminase(SGOT).Results:Results indicated that the methanol extract of *** was non-toxic and hepatoprotective in nature as compared with the petroleum ether *** acetone fraction of methanolic extract also showed similar properties but the subsequent two fractions were ***,the pure compound isolated from the penultimate fraction of methanolic extract was non-toxic and hepatoprotective in *** investigations(SCOT,SCPT) further corroborated these cytological ***:It can be concluded from this study that *** methanol extract,some fractions and pure isolated compound herein exhibit hepatoprotective ***,cytotoxicity recorded in the penultimate fraction and investigation of structural details of pure compound warrants further study.

关键词： Feronia limonia leaf HepG2 Cytotoxicity Hepatoprotective Phytochemical analysis TLC HPLC Hepatoprotective activity SGPT SGOT Isolated compound Spectroscopic assay

Spectrofluorometric assays of Human Collagenase Activity Using Native Collagen and Acetyl-Peptide Substrates

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Advances in Enzyme Research 2015年第1期3卷 19-29页

作者： Valon Ejupi Shpend Dragusha Tsutomu Kabashima Qinchang Zhu Ahmed F. M. El-Mahdy Sheng Yin Takayuki Shibata Masaaki Kai Faculty of Pharmaceutical Sciences Graduate School of Biomedical Sciences Nagasaki University Nagasaki Japan

A selective, sensitive, and convenient assay for human collagenase is required because of its implication in diseases such as rheumatoid arthritis, osteoarthritis, and tumors. Here, a novel assay for human collagenase activity is described in which enzymatic degradation of native collagen or acetyl peptide is determined by using a fluorogenic reaction for oligopeptides. The oligopeptides are quantified spectrofluorometrically with either 3,4-dihydroxyphenylacetic acid or 1,2-dihydroxybenzen reaction in the presence of sodium periodate and sodium borate (pH 7 - 8). These reactions can selectively convert N-terminal Gly-containing oligopeptides and N-terminal Ile-containing oligopeptides to fluorescence (FL) compounds, respectively, but not proteins, acetyl peptides or amino acids. Under optimized conditions using 1.65 μM collagen IV or 1.5 mM Ac-GPQGI- AGQ as substrates, this assay exhibits a proportional relationship between FL intensities and human collagenase-3 (MMP-13) concentrations. It can assay endogenous collagenase activities in several biological samples, such as cultured human cells and cheek tissue.

关键词： Spectrofluorometric assay Human Collagenase Metalloproteinase Native Collagen Acetyl Peptide

Feasibility study on replacing chemical uranium assays with uranium equivalent grades at Husab Uranium Project,Namibia

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Global Geology 2018年第4期21卷 237-244页

作者： CAO Yun LUO Yi LI Zheng Sichuan Institute of Nuclear Geology Chengdu 610052China

Formerly,uranium exploration at the Husab Uranium Project was based on sample chemical uranium assays with little uranium equivalent data( e U) obtained from γ probing. The high cost of performing chemical assays has a significant impact on the overall cost of uranium exploration. Between 2015 and 2016,the Resource Upgrade Project Zone 2 was undertaken at the Husab Project,which identified the reliability and feasibility of γ probing through comparison of chemical uranium assays with uranium equivalent grades. This project analyzed the sources of errors and proposed a method to correct them. Uranium equivalent data from gamma probing can be used to replace the chemical assays to reduce the cost and time,therefore increasing the efficiency for uranium exploration at the Husab Uranium Project.

关键词： comparison chemical uranium assay uranium equivalent grade error replace

Comparative Analysis of Pathogenicity assays and PCR for <i>Listeria monocytogenes</i>Isolated from Animal, Raw Food and Environmental Sources in Korea

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Advances in Microbiology 2013年第2期3卷 191-199页

作者： Sanghun Park Jihun Jung Younghee Oh Jibho Lee Seunghee Ryu Hyowon Jung Sunhee Park Miok Song Gunyong Park Sungmin Choi Sangmi Lee Junghun Kim YoungZoo Chae Byungyeol Jung Myunghun Lee Hyunsoo Kim Seoul Metropolitan Government Research Institute of Public Health and Environment Yangjae-Dong Seocho-Gu Seoul Korea

Results of PCR with oligonucleotide primers were designed from the assembled panel of four potential virulence genes (two of internalin gene and two of transcriptional regulator gene). Most of the isolates including reference strains were reactive by PCR, whereas the other strains (No.80, 81, and 83) isolated from pork, were non-reactive by PCR. In particular, all pork isolates were PCR-negative for two primers (lmo2672 and 2821) sets tested. However, No.82 was positive for lmo1134 primer, and No.84 was positive for lmo2470 of pork isolates. It was observed that all Listeria monocytogenes (L. monocytogenes) penetrate Vero cells, although the invasion efficiency of each strain varied (between 0.5 and 18.9%). When compared in cell assay with PFGE, the results were shown that the mean invasion efficiency for lineage II isolate (2.6%) was significantly lower (ANOVA-test,

关键词： PFGE Vero Cell Listeria monocytogenes in Vitro assay

Cell Survival assays for Individualised Chemotherapy in Primary Glioma Cultures—Colourmetric or Luminescent?

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Neuroscience & Medicine 2011年第4期2卷 376-383页

作者： Timothy P. Dawson Katherine Ashton Aprajay Golash Charles H. G. Davis R. Venkat Iyer Phil Roberts Fred Harris Robert W. Lea 不详

In vitro chemosensitivity testing of short term primary glioma cultures derived from brain biopsies is still in the research phase and has not yet found a place in clinical use. The main reasons for this slow progression are the small amounts of tissue available and the lack of a suitably sensitive assay capable of use in the clinical setting. This study examines whether the MTS and ATP cell survival assays, which determine cytotoxicity via colorimetric and luminescence analysis respectively, could potentially fulfill this role. Primary glioma cultures were tested for chemosensitivity using the MTS and ATP assays and were found to be generally sensitive to cisplatin and paclitaxel but relatively resistant to carmustine and etoposide. For both assays, LD50 values lay in the range 2 - 130 μg/ml but in the vast majority of cases, those obtained by the ATP assay were markedly lower those obtained by the MTS assay. Moreover, at cell numbers less than 2000 in the cases of paclitaxel and carmustine and less than 4500 in the case of cisplatin, these drugs were generally indicated as ineffective against the glioma cultures tested by the MTS assay but effective against these cultures by the ATP assay. These data clearly demonstrate that the ATP assay is more sensitive when estimating small cell numbers generated by primary glioma cultures from brain biopsies and more reliably detects higher kill rates by anticancer drugs. This study also supports the feasibility of using the ATP assay for chemosensitivity testing in a clinical setting.

关键词： ATP assay MTS assay Glioma Primary Tumour Culture In Vitro Chemosensitivity Testing

Development of time-resolved immunofluorometric assays for the detection of house dust mite-allergic IgE in human sera

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Health 2010年第11期2卷 1280-1286页

作者： Ratchanoo Phiphatchaipaisarn Jundee Rabablert Kornkarn Bramarapravati Duangthep Thongdee Nares Wongpitoon Worawan Durongpisitkul Nat Malainual 不详

Dermatophagoides farinae and D. pteronyssinus are the prevalent house dust mites (HDM) in tropical countries and are associated with allergic diseases. This investigation developed a time- resolved immunofluorometric assay (TR-IFMA) for the first time to detect specific IgE antibody in patients with skin prick test positive to HDM but no detectable IgE by other means. Levels of IgE to natural and recombinant HDM allergens were measured by TR-IFMA in 50 HDM-allergic patients and 19 healthy participants compared to sandwich enzyme-linked immunosorbent assay (ELISA). A recombinant allergen, rDerf2, showed a 14 kDa band corresponding to broad range proteins of natural ***-IFMA showed sensitivity lower than 0.35 kUA/l. TR-IFMA employing three HDM antigens showed good correlations with sandwich ELISA at R2 0.93-0.96. TR-IFMA detected HDM IgE in 62, 62, 25 percent of allergic patient serum sample compared to 28, 32, and 22 percent detected by ELISA result using three HDM allergen. TR-IFMA also detected 26.3, 31.6, and 5.3 percent positive samples from 19 healthy participants while ELISA showed 0, 5.3, and 0 percents IgE positive samples. The use of rDerf2 as an HDM allergen for the assay was verified with no statistically different from other HDM allergens. TR-IFMA showed lower detection limit than ELISA and yielded higher sensitivity for serum of people with allergic symptoms with no detectable HDM IgE. It is anticipated that TR-IFMA for HDM-specific IgE detection will play an important role in future diagnosis of HDM allergy in clinical laboratories and for different research purposes.

关键词： House Dust Mites Time-Resolved Immunofluorometric assay Allergy IgE

Immunological-based assays for specific detection of shrimp viruses

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World Journal of Virology 2014年第1期3卷 1-10页

作者： Parin Chaivisuthangkura Siwaporn Longyant Paisarn Sithigorngul Department of Biology Faculty of Science Srinakharinwirot University Bangkok 10110 Thailand Faculty of Environmental Culture and Ecotourism Srinakharinwirot University Bangkok 10110 Thailand

Among shrimp viral pathogens, white spot syndrome virus(WSSV) and yellow head virus(YHV) are the most lethal agents, causing serious problems for both the whiteleg shrimp, Penaeus(Litopenaeus) vannamei, and the black tiger shrimp, Penaeus(Penaeus) monodon. Another important virus that infects P. vannamei is infectious myonecrosis virus(IMNV), which induces the white discoloration of affected muscle. In the cases of taura syndrome virus and Penaeus stylirostris densovirus(Pst DNV; formerly known as infectious hypodermal and hematopoietic necrosis virus), their impacts were greatly diminished after the introduction of tolerant stocks of P. vannamei. Less important viruses are Penaeus monodon densovirus(Pm DNV; formerly called hepatopancreatic parvovirus), and Penaeus monodon nucleopolyhedrovirus(Pemo NPV; previously called monodon baculovirus). For freshwater prawn, Macrobrachium rosenbergii nodavirus and extra small virus are considered important viral pathogens. Monoclonal antibodies(MAbs) specific to the shrimp viruses described above have been generated and used as an alternative tool in various immunoassays such as enzyme-linked immunosorbent assay, dot blotting, Western blotting and immunohistochemistry. Some of these MAbs were further developed into immunochromatographic strip tests for the detection of WSSV, YHV, IMNV and Pemo NPV and into a dual strip test for the simultaneous detection of WSSV/YHV. The strip test has the advantages of speed, as the result can be obtained within 15 min, and simplicity, as laboratory equipment and specialized skills are not required. Therefore, strip tests can be used by shrimp farmers for the pond-side monitoring of viral infection.

关键词： Shrimp viruses Immunological assay Western blot Immunohistochemistry Immunochromatographic strip test