• The Plant Pathology Journal
• /
• v.20 no.2
• /
• pp.142-146
• /
• 2004

Woody plant tissues contain great amounts of phenolic compounds and polysaccharides. These substances inhibit the activation of reverse transcriptase and/or Taq polymerase in RT-PCR. The commonly used multiple-step protocols using several additives to diminish polyphenolic compounds during nucleic acid extraction are time consuming and laborious. In this study, sodium sulfite was evaluated as an additive for nucleic acid extraction from woody plants and the efficiency of RT-PCR assay of commercial nucleic acid extraction kits and small-scale dsRNA extraction was compared. Sodium sulfite was used as an inhibitor against polyphenolic oxidases and its effects were compared in RNA extraction by commercial extraction kit and small-scale double-stranded RNA (dsRNA) extraction method for RT-PCR. During nucleic acid extraction, addition of 0.5%-1.5%(w/v) of sodium sulfite to lysis buffer or STE buffer resulted in lighter browning by oxidation than extracts without sodium sulfite and improved the RT-PCR detection. When commercial RNA extraction kit was used, optimal concentrations of sodium sulfite were variable according to the tested plant. However, with dsRNA as RT-PCR template, sodium sulfite 1.5% in STE buffer improved the detection efficiency of Apple chlorotic leaf spot virus (ACLSV) and Apple stem grooving virus (ASGV) in fruit trees, and reduced the unspecific amplifications signi-ficantly. Furthermore, when viruses existed at low titers in host plant, small-scale dsRNA extractions were very reliable.

• Tuberculosis and Respiratory Diseases
• /
• v.39 no.2
• /
• pp.159-166
• /
• 1992

Background:Sulfiting agents are widely used as preservatives and antioxidants in foods, beverages and drugs including bronchodilators. There have been reports of sulfite-related reactions such as anaphylaxis, urticaria, angioedema, abdominal discomfortness as well as bronchospasm. Several investigators reported that sulfite-sensitive asthmatic patients comprised from 3.9% to 8.2% of all asthmatic patients and its prevalence was higher in steroid-dependent group than in steroid-independent group. Subjects and Method:We performed oral provocation test with sodium bisulfite and aspirin in 17 asthmatic patients who have experienced aggravation of their symptoms after administration of drugs or foods. All of them were steroid dependent asthmatics. We observed clinical symptoms and steroid requirements from 1 to 18 months. Result:Ten of them showed severe bronchoconstriction after the ingestion of sodium bisulfite (50 to 200 mg) within 30 minutes. Concurrent aspirin intolerance was noted by oral provocation test in four cases (40%). Three of them showed positive responses on skin prick test with sulfite (10 or 100 mg/ml). Mean total eosinophil counts was $844/mm^3$ at asthmatic attack. And there was no significant responses on skin prick test and IgE-RAST to common inhalant allergens. After complete avoidance from sulfite containing foods and drugs as well as antiasthmatic medication for 1 to 18 months, nine of them (90%) could stop or reduce the steroid requirements. ConcIusion:It was suggested that severe steroid dependent and intrinsic type of asthmatic patients should be evaluated for sulfite-sensitivity.

• Journal of the Korean Graphic Arts Communication Society
• /
• v.14 no.2
• /
• pp.37-49
• /
• 1996

The practical lith developer is a hydroquinone solution of moderate alkalinity and low free-sulfite ion concentration with potassium bromide as the restraining agent. In the study on composition of ilth developer to promote high sensitivity, high contrast and long term preservation or use, a new composition of lith developer, namely PK lith developer, was developed and of was found that PK lith developer was superior to the lith developer which was commercially available in terms of contrast, speed and preservation. This paper also describes studies on the addition of hdydrazine to PK lith developer, which have led to further improvements in the system.

• Research in Plant Disease
• /
• v.12 no.2
• /
• pp.139-143
• /
• 2006

Virion captured reverse transcription polymerase chain reaction (VC/RT-PCR) could detect plant virus quickly and accurately. In the VC/RT-PCR, no antibody is needed unlike immuno-captured RT-PCR (IC/RT-PCR) which had been improved method of RT-PCR for plant viruses, and virus nucleic acids can be obtained easily within 30minutes by property of polypropylene PCR tube which is hold and immobilized viral particles on its surface. For the virion capture of Tomato spotted wilt virus (TSWV), the extraction buffer was tested. The optimum macerating buffer for TSWV was 0.01M potassium phosphate buffer, pH 7.0, containing 0.5% sodium sulfite. The viral crude sap was incubated for 30 min at $4^{\circ}C$. The virions in the PCR tubes were washed two times with 0.01M PBS containing 0.05% Tween-20. The washed virions were treated at $95^{\circ}C$ immediately for 1 min containing RNase free water and chilled quickly in the ice. Disclosed virions' RNAs by heat treatment were used for RT-PCR. Dilution end point of $10^{-5}$ from plant's crude sap infected with TSWV showed relatively higher detection sensitivity for VC/RT-PCR. During multiple detection using two or more primers, interference was arisen by interactions between primer-primer and plant species. The result of multiplex RT-PCR was influenced by combinations of primers and the kind of plant, and the optimum extraction buffer for the multiplex detection by VC/RT-PCR should be developed.