Isolation of intact nuclei from Euglena Grancilis.

A method for the isolation of nuclei from the lower eukaryote, Euglena gracilis, utilizing MES buffer at pH 5.5 and sodium meta bisulfite is presented. The method takes advantage of the finding that sodium meta bisulfite reversibly alters the density of the nuclei allowing them to be separated from cytoplasmic fragments and unbroken whole cells. Nuclei are obtained in a yield of 25–37 %, appear intact ultrastructurally, and contain acid-soluble proteins in an amount relatively the same as found in higher cell nuclei.     

Interaction between 2,4-Dichlorophenoxyacetic Acid and Vitamin K in Datura stramonium

Plants of the cross Datura stramonium L. var. stramonium×Datura stramonium L. var. inermis (Juss. ex Jacq.) Timm. were sprayed either with a water solution of the sodium salt of 2,4-dichlorophenoxyacetic acid (2,4-D) or with a combination of 2,4-D (sodium salt) and the vitamin K compound menadione sodium bisulfite in water solution. The plants were found to differ in one of their responses to the treatments, namely the development of necrotic spots on the leaf blades. Many necroses appeared on the plants treated with 2,4-D plus menadione sodium bisulfite while only a few necroses or none at all developed on the plants treated with 2,4-D alone. Furthermore, 2,4-D in combination with menadione sodium bisulfite induced much larger necrotic spots than did 2,4-D by itself. Spraying with menadione sodium bisulfite in water solution did not produce any observable effects upon Datura plants. The great increase in necrotic lesion formation was probably due to an interaction between 2,4-D and menadione sodium bisulfite (or a metabolite of this compound) within leaf cells.     

Effects of phosphatase inhibitors on nuclease activity

Nucleases such as DNase I, which selectively digest chromatin, are inhibited by several commonly used phosphatase inhibitors including sodium bisulfite. Two inhibitors, sodium arsenate and fructose-1,6-diphosphate, did not significantly inhibit nuclease action. Two other effective phosphatase inhibitors, p-chloromercuriphenyl sulfonate and 5,5'-dithiobis(2-nitrobenzoate), can be used during nuclei isolation and then washed out of nuclei before nuclease digestion. Using this procedure, 1mM p-chloromercuriphenyl sulfonate is as effective as 50mM bisulfite in retaining the phosphatase-sensitive mitotic phosphorylations of histones H1 and H3.     

Determination of a Non-methylated Deoxycytidine Residue in the Recognition Site of DNA-methyltransferases

A method for determination of a non-methylated deoxycytidine (dC) residue in the recognition site of 5-cytosine DNA-methyltransferases is suggested. The method is based on treatment of methylated DNA by sodium bisulfite and successive reaction of the thus modified DNA with a repair enzyme, uracil-DNA glycosylase. This method was successfully applied to identify NlaX methyltransferase specificity.     

Synthesis of <Emphasis Type="Italic">gem</Emphasis>-disubstituted taurines by the regioselective ring-opening of 2,2-disubstituted aziridines with sodium bisulfite and sulfite

The regioselectivity of the ring-opening reactions of 2,2-disubstituted aziridines with sodium bisulfite and sulfite showed significant divergence depending on the nucleophiles and the structure of the substituents of the aziridines. 2,2-Dialkylaziridines were specifically attacked on their less substituted carbon atoms with sodium bisulfite, affording 2,2-substituted taurines, while 2,2-disubstituted aziridines with either one or two aryl substituent(s) were attacked specifically on their more substituted carbon atoms with the same nucleophile, giving rise to aromatic 1,1-substituted taurines. Sodium sulfite attacked the less substituted carbon atoms of all aziridines specifically to afford 2,2-disubstituted taurines. The regioselectivity is governed by the nucleophile and the balance between the steric hindrance and the electronic effect. The current method provides an alternative route to the synthesis of 2,2-dialkyltaurines and aromatic gem-disubstituted taurines.     

二氧化硫衍生物对玉米种子萌发、幼苗生长和细胞分裂的影响

研究3种二氧化硫衍生物硫酸钠、亚硫酸钠和亚硫酸氢钠对玉米种子萌发、幼苗生长和细胞分裂的影响。结果表明,一定浓度(0.1 mmol·L^-1)的硫酸盐和亚硫酸盐处理液能促进玉米种子萌发和幼苗生长,但浓度超过2.5 mmol·L^-1时能抑制种子萌发和幼苗生长。较高浓度的3种盐溶液对根尖分生区细胞的分裂具有抑制作用,并导致其中的部分细胞异常,出现细胞核固缩,甚至产生质壁分离。3种二氧化硫衍生物的毒性排序为：亚硫酸氢钠＞亚硫酸钠＞硫酸钠。     

The effects of sodium bisulfite in extraction of PHB by hypochlorite

Summary Molecular weight of PHB becomes low when using a hypochlorite extraction method for PHB separation. This disadvantage was overcome by adding sodium bisulfite which is an anti-oxidant. The molecular weight drop was decreased from 30 % to 14 % by adding sodium bisulfite. It was also found that raising the pH to 12 and the addition of a surfactant resulted in the improvement of the PHB purity.     

Oxidation of 2-mercaptoacetyl-phenylalanyl-leucine,a specific inhibitor ofPseudomonas aeruginosa elastase,in the presence of the bacteria and its prevention

The compound 2-mercaptoacetyl-l-phenylalanyl-l-leucine (HSAc-Phe-Leu) is a specific and potent inhibitor ofPseudomonas aeruginosa elastase with demonstrated potential as a drug for the treatment ofPseudomonas eye infections. The thiol moiety of this compound is lost in the presence of the bacterial at a rate greater than that seen in the absence of the organisms. Tritium-labeled inhibitor was prepared and used to examine whether this accelerated loss is due to the uptake of the inhibitor by the bacteria or due to its modification by some bacterial products. Our results excluded by possibility of uptake and indicated that the loss of the thiol resulted from its oxidation to the inactive disulfide form. The oxidation reaction is probably catalyzed by a low molecular weight extracellular bacterial product, and is effectively prevented in the presence of sodium bisulfite. It is suggested that HSAc-Phe-Leu preparations used for further investigation of the inhibitor's therapeutic potential should include antoxidants such as sodium bisulfite to provide maximal inhibitory capacity.     

Fate of aflatoxins B1 and G1 residues during biscuit processing

Effect of biscuit processing on the destruction of aflatoxins B₁ and G₁ with and/or without some commonly leavening agents used namely sodium bicarbonate, ammonium bicarbonate and sodium bisulfite and sodium chloride. It was found that mixing step reduced the concentration of aflatoxins B₁ and G₁ by 80.7% and 82.7%, while the effect of baking step being 28.9% and 21.5%. The effect of mixing was found to be more pronounced than that baking step. The highest destruction effect on aflatoxin B1 was observed by adding a mixture composed of sodium and ammonium bicarbonate and sodium bisulfite followed by sodium chloride, sodium bisulfite, ammonium bicarbonate and/or sodium bicarbonate alone, where the reduction values of toxin after mixing were 93.4,91.9,91.7, 88.8 and 86.6% respectively, while the baking effect ranged 17.2 to 34.5% in the presence of different leaving agents added. Concerning aflatoxin G1; the highest destructive effect of toxin was adsorbed by adding a mixture of sodium and ammonium bicarbonate and sodium bisulfite followed by sodium bisulfite, sodium chloride, ammonium bicarbonate and/or sodium bicarbonate alone since the destruction values of such toxin after mixing were 96.2%, 92.8%, 92.6%, 89.0% and 87.7% respectively, while the baking effect ranged 20.9 to 34.5% in all leavening agents added.     

Acidic conditions enhance bactericidal effects of sodium bisulfite on Helicobacter pylori

BACKGROUND: The Brucella broth medium, which is often used for the cultivation of microaerobic bacteria including Helicobacter pylori. It contains sodium bisulfite to decrease oxygen content in the medium. The growth of H. pylori, however, is inhibited by sodium bisulfite. In this study, the effect of sodium bisulfite was compared with several antioxidants and quantified under acidic conditions, mimicking the gastric environment. METHODS: Growth of H. pylori in the presence of several antioxidants was evaluated at OD655 nm. Effect of sodium bisulfite on H. pylori under acidic conditions was evaluated by measuring colony forming units (cfu). RESULTS: Under neutral conditions, sodium bisulfite was a more potent suppressor of H. pylori. Resveratrol, a polyphenol found in wine, exhibited the most potent inhibitory activity. To quantify the effect of sodium bisulfite on H. pylori under acidic conditions, the bacteria were grown at 37 degrees C for 30 minutes in 0.15 mol/l HCl/KCl (pH 2.0) with or without urea and sodium bisulfite. Sodium bisulfite (0.5 mmol/l) did not affect the viability at neutral pH 7.0, however, it killed H. pylori under acidic conditions, even if urea, the key substance enabling H. pylori to survive under acidic conditions, was present. The bacteria, which had been incubated under acidic conditions in the presence of urea, could survive a subsequent 30 minute-incubation at pH 2.0 without urea. Presence of sodium bisulfite, however, in the subsequent 30 minute-incubation, killed the bacteria. CONCLUSIONS: The bactericidal effect of sodium bisulfite on H. pylori was greater under acidic conditions and independent of urease activity.     

The protein composition of Friend cell nuclear matrix stabilized by various treatments. Different recovery of nucleolar proteins B23 and C23 and nuclear lamins

Summary— Using two-dimensional polyacrylamide gels stained with Coomassie blue we have studied the protein composition of the nuclear matrix obtained from mouse erythroleukemic nuclei kept at O°C throughout the isolation procedure to prepare the high ionic strength resistant fraction (control matrix) or stabilized in vitro or in vivo by different procedures prior to subfractionation (ie 37°C incubation of isolated nuclei; sodium tetrathionate exposure of purified nuclei; heat shock of intact cells). When the matrix obtained from 37°C incubated nuclei was compared with the control matrix, striking differences in the polypeptide pattern were seen if the protein was obtained in both cases from an equivalent number of nuclei. On the other hand, if the same amount of protein for both the samples was applied to the gels the differences were less evident. Sodium tetrathionate stabilization of isolated nuclei and heat shock of intact cells produced a matrix protein pattern that was very similar and differed from that of the in vitro heat-exposed matrix. Using specific polyclonal antisera, we demonstrate that nucleolar proteins B23/numatrin and C23/nucleolin were very abundant in the matrix obtained from chemically-treated nuclei or in vivo heat-stabilized nuclei but were recovered in very small amounts (B23) or completely absent (C23) in the matrix prepared from nuclei heated to 37°C in vitro. Differences were seen also in the recovery of nuclear lamins, and especially lamin B, that was poorly represented in the sodium tetrathionate-stabilized matrix. The results demonstrate that in mouse erythroleukemia cells the increased recovery of nuclear matrix protein that is seen after in vitro heating of isolated nuclei is predominantly due to an additional recovery of the same types of polypeptides that are detected also in the absence of such a treatment. The data also indicate that in vivo heat shock of intact cells produces a nuclear matrix protein pattern that is more similar to the pattern seen after stabilization of purified nuclei with sodium tetrathionate and differs significantly from that obtained by exposing nuclei to 37°C in vitro, unlike to that what previous reports have indicated.     

Cytogenetic damage induced in human lymphocytes by sodium bisulfite.

The frequencies of chromosomal aberrations (CA), sister-chromatid exchanges (SCE), and micronuclei (MN) in human blood lymphocytes exposed to sodium bisulfite (sulfur dioxide) at various concentrations ranging from 5 x 10(-5) M to 2 x 10(-3) M in vitro were studied. It was shown that sodium bisulfite (NaHSO3 and Na2SO3, 1:3 M/M) caused an increase in SCE and MN in human blood lymphocytes in a dose-dependent manner, and also induced mitotic delays and decreased mitotic index. For CA, our results indicated that sodium bisulfite induced an increase of chromatid-type aberrations in lymphocytes from three of four donors in a dose-dependent manner. The chemical at low concentrations induced chromatid-type aberrations, but not chromosome-type aberrations; high concentrations induced both chromatid- and chromosome-type aberrations. No cytogenetic damage in human lymphocytes was induced by sodium sulfate. The results have confirmed that sulfur dioxide is a clastogenic and genotoxic agent.     

Cleavage of the glycosidic linkage of pyrimidine ribonucleosides by the bisulfite-oxygen system

When a solution containing 2 mM uridine, 20 mM sodium bisulfite, 0.1 mM MnCl₂, and 100 mM sodium phosphate buffer of pH 7.0 was incubated aerobically at 37° or 0°, partial cleavage of the glycosidic linkage of uridine took place. About 20% of the uridine was converted to uracil by the incubation for 4 hrs. Cytosine was produced from cytidine by similar treatment with bisulfite. These reactions were caused by free radicals generated by Mn²⁺-catalyzed autoxidation of bisulfite. Glycosidic bond cleavage by the bisulfite-oxygen system was not detected for adenosine, AMP, guanosine, GMP, thymidine, TMP, deoxyuridine, dCMP, dAMP, and dGMP. When poly(U) and poly(C) were treated with 20 mM sodium bisulfite in the same manner, chain fission of the polymer occurred as judged by the elution-pattern change in gel filtration through Sephadex columns. No change in the elution pattern was observed for bisulfite-treated poly(A), poly(U)· poly(A) or tRNA.     

Procedure for increased recovery of Campylobacter jejuni from inoculated unpasteurized milk.

Different treatments were applied to Campylobacter jejuni-inoculated unpasteurized milk to identify means of enhancing the survival of the organism in refrigerated (4 degrees C) samples. The greatest survival occurred in milk supplemented with 0.01% sodium bisulfite and held under an atmosphere of 100% nitrogen (bisulfite-nitrogen), in most instances allowing isolation of C. jejuni from highly contaminated milk 15 or more days longer than from unsupplemented milk held in air (21% oxygen). Although a larger amount of Campylobacter was consistently recovered from milk treated with bisulfite-nitrogen, similar isolation rates (qualitative) resulted from milk stored in air and supplemented with 0.01% sodium bisulfite and 0.15% sodium thioglycolate when analyzed within 12 days after sampling. Milk samples to be transported and assayed at a later date would best be held refrigerated (4 degrees C) and supplemented with 0.01% sodium bisulfite and either 0.15% sodium thioglycolate or an atmosphere of 100% nitrogen.     

Prevention of interphase death in rat thymocytes by bisulfite

The effect of sodium bisulfite, a specific inhibitor of chromatin proteolysis, on radiation damage in rat thymocytes in vitro was examined. Rat thymocytes irradiated with 1 kR X rays in vitro were incubated at 37 degrees C with 10 mM glucose for 4 to 6 hr. During that time development of interphase death as judged by erythrosin B uptake, release of low molecular weight DNA (free DNA), and reduction in cell size was measured. Sodium bisulfite added to the cells at the beginning of incubation exerted a marked preventive effect on radiation damage. The effect was enhanced with increasing concentration of bisulfite from 0.25 to 2 mM. The effect of bisulfite was reversible; i.e., removal of bisulfite from the cells resulted in the reappearance of the radiation damage.     

Procedure for increased recovery of Campylobacter jejuni from inoculated unpasteurized milk

Different treatments were applied to Campylobacter jejuni-inoculated unpasteurized milk to identify means of enhancing the survival of the organism in refrigerated (4 degrees C) samples. The greatest survival occurred in milk supplemented with 0.01% sodium bisulfite and held under an atmosphere of 100% nitrogen (bisulfite-nitrogen), in most instances allowing isolation of C. jejuni from highly contaminated milk 15 or more days longer than from unsupplemented milk held in air (21% oxygen). Although a larger amount of Campylobacter was consistently recovered from milk treated with bisulfite-nitrogen, similar isolation rates (qualitative) resulted from milk stored in air and supplemented with 0.01% sodium bisulfite and 0.15% sodium thioglycolate when analyzed within 12 days after sampling. Milk samples to be transported and assayed at a later date would best be held refrigerated (4 degrees C) and supplemented with 0.01% sodium bisulfite and either 0.15% sodium thioglycolate or an atmosphere of 100% nitrogen.     

亚硫酸氢钠测序法检测水稻FIE基因CpG岛甲基化状态

建立了适用于水稻基因组特定基因甲基化检测的亚硫酸氢钠测序法,并利用此方法对FIE2A基因CpG岛部分片段的甲基化差异进行了研究。采用CTAB法提取水稻叶片和胚乳细胞的基因组DNA,经亚硫酸氢钠化学修饰后,针对已修饰的FIE基因序列设计特异引物并结合巢式PCR扩增,TA载体克隆、测序,最后对测序结果进行分析。结果表明巢式PCR能够增加特异性产物的产生,FIE基因CpG岛在对称的CG和CNG位点甲基化水平较高,而在非对称CNN位点甲基化水平最低,此外在叶片中的平均甲基化水平较高。由此表明本研究建立的亚硫酸氢钠测序法适用于水稻基因组特定基因甲基化状态的检测。     

Inactivation of enteroviruses by ascorbic acid and sodium bisulfite.

Poliovirus type 1, coxsackievirus type A9, and echovirus type 7 were inactivated by sodium bisulfite and ascorbic acid. Inactivation rates depended upon concentration, temperature, and pH. RNA infectivity was lost during inactivation; the capsid was also altered by these inactivating agents, as determined by enzyme sensitivity assays and by tests of adsorption to cells. Structural modifications of the virus particles were not identical, suggesting that the mechanism of inactivation by ascorbic acid differs from that of sodium bisulfite.     

Inactivation of enteroviruses by ascorbic acid and sodium bisulfite.

Poliovirus type 1, coxsackievirus type A9, and echovirus type 7 were inactivated by sodium bisulfite and ascorbic acid. Inactivation rates depended upon concentration, temperature, and pH. RNA infectivity was lost during inactivation; the capsid was also altered by these inactivating agents, as determined by enzyme sensitivity assays and by tests of adsorption to cells. Structural modifications of the virus particles were not identical, suggesting that the mechanism of inactivation by ascorbic acid differs from that of sodium bisulfite.     

亚硫酸氢钠在低光强下对叶绿体循环光合磷酸化的促进作用

我们在70年代曾发现喷洒低浓度的亚硫酸氢钠能提高多种作物叶片的光合作用速率(沈允钢等1980)。近年来,谭实和沈允钢(1987)观察到喷洒低浓度的亚硫酸氢钠不但能增加作物叶片的光合作用速率,而且同时增加其呼吸作用速率,并初