Detection of Southern Rice Black-Streaked Dwarf Virus Using Western Blotting With P6

The southern rice black-streaked dwarf virus (SRBSDV) is a severe threat to the yield and quality of rice products worldwide. Traditional detection methods for diagnosing SRBSDV infection show several false positives and thus provide inaccurate findings. However, Western blotting (WB) can precisely solve this problem. In this study, P6—a viral RNA-silencing suppressor—was expressed and purified in vitro. Two polyclonal P6 antibodies were obtained and quantified by enzyme-linked immunosorbent assay and WB. Subsequently, WB was performed using the P6 antibodies to identify SRBSDV antigens derived from the suspected rice samples collected from nine districts in Guizhou, China. The assay results showed that Libo, Pingtang, Huishui, Dushan, and Anshun districts had experienced an SRBSDV outbreak. The virus content in the sampled rice tissues was quantified by WB. Our results revealed that SRBSDV mainly accumulated in rice stems rather than rice leaves. Thus, the findings of our study show that the SRBSDV P6 antibody can be used in WB for detecting and monitoring SRBSDV infection in infected rice plants.

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A Novel Mouse Monoclonal Antibody C42 against C-Terminal Peptide of Alpha-1-Antitrypsin

International Journal of Molecular Sciences ◽

10.3390/ijms22042141 ◽

2021 ◽

Vol 22 (4) ◽

pp. 2141

Author(s):

Srinu Tumpara ◽

Elena Korenbaum ◽

Mark Kühnel ◽

Danny Jonigk ◽

Beata Olejnicka ◽

...

Keyword(s):

Monoclonal Antibody ◽

Flow Cytometry ◽

Western Blotting ◽

Complex Mixture ◽

Immunoglobulin M ◽

Confocal Laser ◽

Enzyme Linked Immunosorbent Assay ◽

Dot Blot

The C-terminal-fragments of alpha1-antitrypsin (AAT) have been identified and their diverse biological roles have been reported in vitro and in vivo. These findings prompted us to develop a monoclonal antibody that specifically recognizes C-36 peptide (corresponding to residues 359–394) resulting from the protease-associated cleavage of AAT. The C-36-targeting mouse monoclonal Immunoglobulin M (IgM) antibody (containing κ light chains, clone C42) was generated and enzyme-linked immunosorbent assay (ELISA)-tested by Davids Biotechnologie GmbH, Germany. Here, we addressed the effectiveness of the novel C42 antibody in different immunoassay formats, such as dot- and Western blotting, confocal laser microscopy, and flow cytometry. According to the dot-blot results, our novel C42 antibody detects the C-36 peptide at a range of 0.1–0.05 µg and shows no cross-reactivity with native, polymerized, or oxidized forms of full-length AAT, the AAT-elastase complex mixture, as well as with shorter C-terminal fragments of AAT. However, the C42 antibody does not detect denatured peptide in SDS-PAGE/Western blotting assays. On the other hand, our C42 antibody, unconjugated as well as conjugated to DyLight488 fluorophore, when applied for immunofluorescence microscopy and flow cytometry assays, specifically detected the C-36 peptide in human blood cells. Altogether, we demonstrate that our novel C42 antibody successfully recognizes the C-36 peptide of AAT in a number of immunoassays and has potential to become an important tool in AAT-related studies.

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Frecuencias de corte y niveles de fertilización nitrogenada en rendimiento y calidad del forrraje de morera (Morus sp.), en Cuyuta, Guatemala

Agronomía Mesoamericana ◽

10.15517/am.v3i0.25204 ◽

2016 ◽

Vol 3 ◽

pp. 48

Author(s):

Carlos Rodríguez ◽

Juan A. Quiñones ◽

Rodrigo Arias

Keyword(s):

Dry Matter ◽

Ground Level ◽

Dry Matter Digestibility ◽

Yield And Quality ◽

Whole Plant ◽

Dual Purpose Cattle ◽

Whole Plants ◽

Cutting Height

The trial was conducted at the Centro de Producción Agrícola of ICfA in Cuyuta, Escuintla-Guatemala in order to generate information on the yield and quality of edible mulberry (Morus sp.) roughage treatments consisted of three harvest frequencies (6; 9 and 12 weeks and three fertilization levels of nitrogen (0.40 and 80 kg/ha). A complete randomized block experimental design, with a factorial arrangement (3x3) was used. The cutting height was 0.3 m above the ground level with two sequencies: from August 2nd to September 13 th and from September 13th to December 6th, 1990. In both cases, the 12 week frequency cutting and 80 kg of N were superior to the others (P<=0.01), yielding 6.87 and 6.15 t/ha of dry matter respectively. The yields at 9 weeks were statistically higher than those at 6 weeks. The highest protein percentage of the whole plant, leaves and stalks were produced at 6 weeks. The dry matter digestibility values in vitro showed little variability among treatments, with averages for whole plants, leaves and stalks of 65, 91 and 41 %, respectively. The preceeding data suggests that the mulberry has an excellent potential as a balanced supplement (protein and energy) for dual purpose cattle on the Southern coast of Guatemala.

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YIELD AND QUALITY OF SOME KALE-CORN COMBINATIONS

Canadian Journal of Plant Science ◽

10.4141/cjps80-119 ◽

1980 ◽

Vol 60 (3) ◽

pp. 807-811 ◽

Cited By ~ 2

Author(s):

R. S. FULKERSON

Keyword(s):

Protein Content ◽

Corn Stover ◽

Crude Protein ◽

Dry Matter ◽

In Vitro Digestibility ◽

Crude Protein Content ◽

Yield And Quality ◽

Row Width

Midas marrowstem kale (Brassica oleracea L.) was grown in different row width associations with United 106 corn (Zea maize L.) in two studies and ensiled in different moisture blends with corn stover in another. Highest dry matter yields were obtained where a single row of kale was grown at 30 cm to the side of a corn row. This combination also provided the lowest moisture content feed and the highest in vitro digestibility and crude protein content. Changing the corn row width had no significant effect upon yield, plant height, in vitro digestibility, kale leaf or corn ear content. Blending kale with corn stover to provide a silage of about 70% moisture increased the digestibility and protein content of the feed and provided a silage that kept well in storage.

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Yield and quality of forage from intercrops of barley and annual ryegrass

Canadian Journal of Plant Science ◽

10.4141/cjps92-016 ◽

1992 ◽

Vol 72 (1) ◽

pp. 163-172 ◽

Cited By ~ 5

Author(s):

D. J. Thompson ◽

D. G. Stout ◽

Z. Mir ◽

T. Moore

Keyword(s):

Spring Barley ◽

Forage Yield ◽

Annual Ryegrass ◽

Hordeum Vulgare L ◽

Potential Yield ◽

Barley Cultivars ◽

Yield And Quality ◽

South Central

Three spring barley (Hordeum vulgare L.) and four annual ryegrass (Lolium multiflorum Lam.) types were intercropped to evaluate the potential yield and quality of forage which can be produced under irrigation in southern interior B.C. All treatments were intercrops; when barley cultivar effects are described they are averaged over the ryegrasses and vice versa. Barley cultivars differed in grain maturity. Ryegrass cultivars included diploid and tertraploid Italian and Westerwolds types. Intercrops including late grain maturing barley cultivars (Samson and Virden) increased the yield of the first silage cut (both by 25% over 2 yr) compared to Diamond, a medium-maturing cultivar adapted to the area. Intercrops containing the semi-dwarf barley, Samson, produced more digestible forage including higher in vitro digestible dry matter (IVDDM) and lower ADF and lignin. Annual ryegrass yield in the first cut intercropped with Samson was almost twice that with either Diamond or Virden, showing that Samson barley is less competitive. Second-cut yield (ryegrass regrowth) was greater for tetraploid than diploid annual ryegrasses. Yields of fall pasture (cuts 3 and 4) were similar among ryegrass cultivars. Cuts 2 and 3 (ryegrass only) of Italian ryegrasses had superior quality (higher IVDDM and protein; and lower ADF, lignin, and NDF) to Westerwolds ryegrassses, but all cultivars had similar quality in the late fall (Cut 4). It was concluded that a range of barley and annual ryegrass cultivars can be successfully intercropped to produce forage in south central B.C.Key words: Intercropping, barley annual ryegrass, forage yield and quality

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Mechanisms Underlying H2O2-Evoked Carbonyl Modification of Cytoskeletal Protein and Axon Injury in PC-12 Cells

Cellular Physiology and Biochemistry ◽

10.1159/000491975 ◽

2018 ◽

Vol 48 (3) ◽

pp. 1088-1098 ◽

Cited By ~ 2

Author(s):

Xiejun Zhang ◽

Zongyang Li ◽

Qiusheng Zhang ◽

Lei Chen ◽

Xianjian Huang ◽

...

Keyword(s):

Oxidative Stress ◽

Western Blotting ◽

Axonal Injury ◽

Cytoskeletal Proteins ◽

Enzyme Linked Immunosorbent Assay ◽

H2o2 Treatment ◽

Pc 12 Cells ◽

Axonal Dysfunction ◽

Β Tubulin

Background/Aims: To investigate the mechanism that enables oxidative stress and cytoskeleton protein carbonylation to contribute to axonal dysfunction in traumatic brain injury (TBI). Methods: We created an in vitro model of neuronal oxidative damage by exposing a neuron-like cell line (PC-12) to different concentrations (100 μM, 200 μM, and 300 μM) of H2O2 for 24 h or 48 h. Carbonyl modification of cytoskeletal proteins (β-actin and β-tubulin) and its impact on β-actin/β-tubulin filament dynamics were determined by enzyme-linked immunosorbent assay, immunostaining, and western blotting. Depolymerization of β-actin/β-tubulin filaments was evaluated using the monomer/polymer ratio of each protein via western blotting. Phosphorylation of the neurofilament heavy chain (P-NFH) was used as an axonal injury marker and detected by immunostaining. Results: Our results showed that H2O2 treatment led to increased oxidative stress in PC-12 cells, as indicated by the increased generation of malondialdehyde and 8-hydroxydeoxyguanosine and decreased intracellular glutathione levels. H2O2 treatment also increased carbonyl modification of total proteins and cytoskeleton proteins β-actin/β-tubulin, which occurred concurrently with the suppression of proteasome activity. Moreover, H2O2 treatment increased the generation of the axonal injury marker P-NFH, and depolymerization of the β-actin/β-tubulin filaments was indicated by increased monomer/polymer ratios of each protein. Lastly, overexpression of the proteasome β5 subunit in PC-12 cells significantly reduced the H2O2-induced accumulation of carbonylated β-actin/ β-tubulin, P-NFH, and β-actin/β-tubulin depolymerization. Conclusions: We concluded that carbonylation of cytoskeleton proteins could lead to depolymerization of their filaments and axonal injury, and proteasome suppression contributes to the accumulation of carbonylated proteins under oxidative conditions.

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Preparation and Property of Antibacterial Nucleus with the Combination of Nano-Silver and Antibiotics

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.905.282 ◽

2022 ◽

Vol 905 ◽

pp. 282-287

Author(s):

Jie Sun ◽

Le Xin Zhou ◽

Xiao Hong Li ◽

Ming Xia

Keyword(s):

Pathogenic Bacteria ◽

Orthogonal Experiment ◽

Survival Rates ◽

Nuclear Transplantation ◽

Infection Period ◽

Yield And Quality ◽

Inhibition Zones ◽

Prophylactic Method

To improve the yield and quality of pearls in freshwater pearl culture and the survival rates after nucleus implanting surgery, pearl farmers used artificial pearl nuclear transplantation techniques to raise pearls. To address the common problem of Staphylococcus aureus and Escherichia coli infection in oyster farming, a new prophylactic method by using compound antibiotics to prepare the medicine coated pearl nucleus was put forward based on existing research results of the nanosilver antibacterial nucleus. Single-factor experiment, multi-factor experiment, orthogonal experiment, SPSS analysis of variance was used to optimize the antibacterial formulation on the assumption that the contaminated probability of these two pathogenic bacteria was the same. The result showed that the optimal ratio of compound antibiotics was 0.0075g/ml of the flavomycin solution and 0.01g/mL of the terramycin solution; the inhibition zones diameter of both pathogenic bacteria was more than 2.6cm in vitro, which was higher than the nanosilver antibacterial nucleus of 0.9cm in vitro. Indicating that the addition of this compound antibiotic formula for the nanosilver antibacterial nucleus could reduce the usage of antibiotics under the premise of maintaining antibacterial effectiveness, and could preferably inhibit the pathogenic microorganisms in the postoperative infection period. This also indicates that compound antibiotics coated antibacterial nanosilver nucleus would be applied more widely.

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Bioefficacy of rhizospheric fungal isolates against wilt and root-rot pathogens of Pea

International Journal of Agricultural Invention ◽

10.46492/ijai/2020.5.1.20 ◽

2019 ◽

Vol 5 (01) ◽

pp. 133-137

Author(s):

Rashmi S. Nigam ◽

R. U. Khan ◽

Reshu Singh ◽

Joginder Singh

Keyword(s):

Root Rot ◽

Pythium Ultimum ◽

Culture Technique ◽

Dual Culture ◽

Yield And Quality ◽

Fungal Isolates ◽

Antagonistic Potential ◽

Production Areas

Pea (Pisum sativum) is an important leguminous crop in many countries including India. Wilt and root rot of pea is an important and widespread disease that often causes significant reduction in the yield and quality of harvested peas throughout the production areas. It is the most important and widespread disease of pea grown in relatively dry and warm area. In-vitro effectiveness of various antagonistic fungal isolates namely T. harzianum (Th1, Th2, Th3, Th4 and Th5) was evaluated against Fusarium oxysporum f. sp. pisi, Rhizoctonia solani and Pythium ultimum by dual culture technique on potato dextrose agar. According to the observation recorded after 5 days, all the rhizospheric fungal isolates evaluated for their antagonistic potential against wilt and root-rot pathogens, exhibited significant effect on radial growth inhibition of pathogens in comparison to control. Among the fungal isolates, Th3 and Th5 of T. harzianum proved to be most effective in reducing the growth of F. oxysporum f. sp. pisi, R. solani and P. ultimum. It was worthy to note that all rhizospheric fungal isolates visualized an increase in their antagonistic potential over the period of time in subsequent hours of inoculation.

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Electronic cigarette liquid substances propylene glycol and vegetable glycerin induce an inflammatory response in gingival epithelial cells

Human & Experimental Toxicology ◽

10.1177/0960327120943934 ◽

2020 ◽

Vol 40 (1) ◽

pp. 25-34

Author(s):

A Beklen ◽

D Uckan

Keyword(s):

Epithelial Cells ◽

Propylene Glycol ◽

Western Blotting ◽

Electronic Cigarettes ◽

Periodontal Diseases ◽

Biological Response ◽

Electronic Cigarette ◽

Enzyme Linked Immunosorbent Assay ◽

Gingival Epithelial Cells

Information on the effects of propylene glycol (PG) and vegetable glycerin (VG) and on cytotoxicity and subsequent activation of the biological mediators is limited in periodontal diseases. This study analyzes the effect of unflavored PG/VG alone or in combination with nicotine on gingival epithelial cells. The cells were exposed to different PG/VG (± nicotine) concentrations for 24 h and cytotoxicity was evaluated by calorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromid assay. The expressions of interleukin (IL)-6, IL-8, and matrix metalloproteinases (MMPs)-9 were measured using an enzyme-linked immunosorbent assay and a western blotting. Stimulation with PG/VG mixtures reduced cell viability compared to nonexposed controls ( p < 0.05). Adding PG/VG increased the levels of IL-6, IL-8, and MMP-9, and the amount of PG had more biological impact compared to the VG amount. The nicotine augmented this effect compared to its nicotine-free counterparts. In western blotting result, MMP-9 was clearly activated in almost all samples. These findings suggest that the main constituents PG/VG are cytotoxic and able to induce biological response in gingival cells in vitro. Despite being advertised as less harmful than conventional cigarettes, electronic cigarette liquid pose certain risks on periodontal cells. Awareness about the effects of electronic cigarettes on periodontal diseases must be increased.

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Intrapopulation Antagonism Can Reduce the Growth and Aggressiveness of the Wheat Head Blight Pathogen Fusarium graminearum

Phytopathology ◽

10.1094/phyto-09-19-0341-r ◽

2020 ◽

Vol 110 (4) ◽

pp. 916-926

Author(s):

Martha M. Vaughan ◽

Todd J. Ward ◽

Susan P. McCormick ◽

Nathane Orwig ◽

William T. Hay ◽

...

Keyword(s):

Fusarium Graminearum ◽

Management Practices ◽

Toxin Production ◽

Antagonistic Effect ◽

Head Blight ◽

Yield And Quality ◽

Pathogen Populations ◽

Moderately Resistant

Fusarium graminearum is a causal agent of Fusarium head blight (FHB), a disease that reduces yield and quality of cereal crops and contaminates grain with mycotoxins that pose health risks to humans and livestock. Interpopulation antagonistic interactions between isolates that produce different trichothecene mycotoxins can reduce FHB in wheat, but it is not known if interactions between isolates with a shared population identity that produce the same trichothecenes have a similar effect. Using isolates from the predominant F. graminearum populations in North America (NA1 and NA2), we examined intrapopulation interactions by comparing growth, disease progression, and toxin production of individual isolates with multi-isolate mixes. In vitro, mycelial growth was significantly greater when most NA1 and NA2 isolates were cultured individually versus when cultured as a mixture of isolates from the same population. In susceptible wheat Norm, FHB generally progressed faster in heads inoculated with an individual isolate versus a multi-isolate mixture, but the antagonistic effect of intrapopulation interactions was more pronounced for NA1 than NA2 isolates. By contrast, in moderately resistant wheat Alsen, mixtures of isolates from either population caused obvious reductions in FHB development. Mycotoxin contamination was not consistently affected by intrapopulation interactions and varied depending on the interacting isolates from either population. Our results indicate that antagonistic intrapopulation interactions can influence FHB in controlled environmental conditions. Understanding if the regional composition of pathogen populations similarly influences FHB in the field could improve disease forecasting and management practices.

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