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1.
以水泡性口炎病毒(VSV)为指示病毒,考察了低pH孵放不同时间对低温乙醇法生产的静注丙球(IVIG)中VSV的灭活效果,并对不同厂家及不同批号IVIG中病毒灭活情况进行了比较。结果表明,液体IVIG在PH4.1±0.3,20-25℃,孵放21天可灭活VSV达6Logs以上(即低于实验检测限),但不同厂家及不同批号的IVIG在病毒灭活的发生上有所不同 相似文献
2.
在人纤维蛋白原制备工艺中增加S/D处理灭活病毒步骤,TNBP和Tween80终浓度分别为0.3%和1%,在25℃处理6小时能有效灭活指示病毒VSV(〉3.75Log)、Sindbis(〉4.46Log)、HIV(〉3.67Log),盲传三代未检出病毒 相似文献
3.
将含有鸡传染性支气管炎病毒 S1 基因c D N A 的重组转移质粒p S X I V V I+ X3 S1 . Holte 和p S X I V V I+ X3/4 S1 . Holte 分别与粉纹夜蛾核型多角体病毒 Tn N P V S V I- G D N A( O C C- ,gal+ ) 共转染草地夜蛾( Sf9) 细胞,经空斑纯化得到重组病毒 Tn N P V( X3) S1 . Holte O C C+ 和 Tn N P V( X3/4) S1 . Holte O C C+ 。将重组毒株分别感染 Tn5 B1 细胞,并进行 S D S P A G E 与 Westernblot 检测。结果表明, Tn N P V( X3/4) S1 . Holte O C C+ 在感染的细胞中高效表达了 S1 蛋白, S D S P A G E 凝胶薄层色谱分析结果显示,感染病毒后72 h S1 蛋白的表达量占细胞内总蛋白量的35 .8 % ,而 Tn N P V( X3) S1 . Holte O C C+ 感染的细胞内检测不出 S1 蛋白。经分析认为这一差异主要来自 S1 基因翻译起始位点及其附近的周围环境。 相似文献
4.
静脉注射免疫球蛋白制备中的病毒灭活 总被引:1,自引:0,他引:1
在静脉注射免疫球蛋白(IVIG)的制备中,采用有机溶剂结合表面活性剂(S/D)处理或60℃10小时液态加热(巴氏灭活法)的方法对组分Ⅱ(IgG)进行病毒灭活处理,灭活效果用指示病毒进行了评价,结果表明:S/D法可有效灭活脂包膜病毒VSV和Sindbis,巴氏灭活法则对上述病毒以及Vaccinia和Echo病毒均有较好的灭活作用。经病毒灭活处理的免疫球蛋白,在理化及生物学特性上基本未受到不利影响,用两种方法分别处理组分Ⅱ后制备的IVIG,其主要特性指标均符合该制品的有关质量规定。 相似文献
5.
麻蝇幼虫肠道蛋白酶BGP的分离纯化及性质 总被引:1,自引:0,他引:1
棕尾别麻蝇幼虫肠液经SDS-PAGE后,X光片显影,呈现两条蛋白酶活性带.IEF后,两条蛋白酶活性带的等电点分别为pH7.7和6.8.麻蝇幼虫肠液经55%~75%硫酸铵沉淀,以及连续两次制备等电聚焦,分离纯化出等电点约为pH7.7,分子量约为35kD的蛋白酶BGP.该酶能分解酪蛋白和类胰蛋白酶专一底物Bz-Phe-Val-ArgNA,不能分解弹性蛋白酶专一底物elastin-CongoRed和类胰凝乳蛋白酶专一底物Suc(Ala)2Pro-PheNA.SBBI,Leupeptin和PMSF能强烈抑制其活性.专一底物和抑制剂的结果表明,BGP是一种类胰蛋白酶.其最适反应温度为50℃,最适作用pH为8.5.不耐高温,50℃保温30min活性急剧下降.Hg2+,Zn2+和Cu2+能抑制酶活性.Ca2+,Mg2+对酶无激活作用,EDTA无抑制作用. 相似文献
6.
从假菠萝麻叶汁中用乙醇分部沉淀法得到一种蛋白酶,对酪蛋白有强烈的水解活性,经Sephadex G-100柱层析和SP-Sephadex C-50离子交换柱层析等步骤纯化,可得到六角形结晶。结晶酶液经PAGE圆盘电泳,每条胶柱上只显示一条蛋白带,其活性在pH4。5-10、55℃范围内较稳定,最适pH8.5最适温度50℃,Km(酪蛋白)值为0.185%(W/V)。用SDS-PAGE和Sephadex 相似文献
7.
Ca^2+敏感性微电极在心肌胞浆Ca^2+活度检测中的应用 总被引:1,自引:0,他引:1
改进的中性载体Ca2+敏感性微电极(Ca-ISE)在尖端直径为0.4—0.8μm时仍具有良好特性,可用于心肌胞浆Ca2+活度(αCai)的检测。测得豚鼠右心乳头肌、狗心室肌和浦肯野纤维静息时分别为0.19±0.01(n=22),0.20±0.02(n=11)和0.46±0.07(n=13)μmol·L-1。毒毛旋花苷G3μmol·L-1使静态及动态心肌分别增高0.18±0.02和6.69±2.09μmol·L-1(n=22),并出现触发活动(TA)。100μmol·L-1蝙蝠葛碱可抑制毒毛旋花苷G引起的增高,同时使其TA消失。表明Ca-ISE技术可用于心肌组织TA与的同步检测。 相似文献
8.
系统感染TMV (tobacco m osaic virus)的番茄(Lycopersicon esculentum Mill.)叶胞外蛋白提取液经冰冻干燥浓缩、- 20℃丙酮沉淀、CM-Sephadex C-25离子交换层析、DEAE-Sephadex A-25离子交换层析和Sephadex G-75凝胶层析纯化,获得PAGE均一的β-1,3-葡聚糖酶.SDS-PAGE证明,它包含分子量为36 kD 和27 kD的两个同工酶.以昆布多糖为底物,酶的最适pH 在4.8—5.2之间,在pH 4—8稳定;酶的最适温度在30—40℃之间,在40℃保温1h 后酶活性不变;Km 值为9.2 m g/m L.在系统感染TMV 的番茄叶胞外蛋白提取液中,有分子量为22 kD、27 kD和36 kD的3个β-1,3-葡聚糖酶同工酶 相似文献
9.
10.
通过单向水平变性聚丙烯酰胺凝胶等电聚焦(IEF)、十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS—PAGE)和双向电泳(IEF×SDS—PAGE)分析了红苋R104种子谷蛋白的亚基组成,亚基分子量和等电点分布。谷蛋白的双向电泳图谱可分辨出100多个亚基成分.其主要亚基为:54kD(pI7.15);33kD(pI5.82);31kD(pI6.92;pI6.70;pI6.65);22kD(pI8.34);20kD(pI6.92;pI656);18kD(pI6.92;pI735;pI7.72;pI8.05)。另外,还对IEF方法进行了讨论。 相似文献
11.
《Comparative biochemistry and physiology. A, Comparative physiology》1979,62(1):219-226
- 1. The two hemoglobins, Hb I and II, of the obligate air-breathing catfish,Hoplosternum littorale have been isolated.
- 2. The unfractionated stripped hemoglobin has a high oxygen affinity, a normal alkaline Bohr effect, and a Root effect.
- 3. Both the Bohr and Root effects are enhanced by 1 mM ATP.
- 4. Stripped Hb I has a relatively high oxygen affinity, a reversed Bohr effect between pH 6.0 and 8.0 (Δlog P502DpH> 0), but no Root effect. Addition of 1 mM ATP to Hb I causes a marked reduction in the oxygen affinity, a change to a normal alkaline Bohr effect (Δlog P50ΔpH< 0), but no Root effect.
- 5. Stripped Hb II has a lower oxygen affinity at low pH and a higher oxygen affinity at high pH than does Hb I. Hb II shows a large alkaline Bohr effect which is only slightly increased by 1 mM ATP and a Root effect at low pH which is enhanced by 1 mM ATP.
- 6. The observed rates of O2 dissociation and of CO combination with Hbs I and II show differences which parallel those observed in the oxygen equilibrium measurements.
12.
Sensitization of Listeria monocytogenes to Low pH, Organic Acids, and Osmotic Stress by Ethanol 总被引:1,自引:0,他引:1
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The killing of Listeria monocytogenes following exposure to low pH, organic acids, and osmotic stress was enhanced by the addition of 5% (vol/vol) ethanol. At pH 3, for example, the presence of this agent stimulated killing by more than 3 log units in 40 min of exposure. The rate of cell death at pH 3.0 was dependent on the concentration of ethanol. Thus, while the presence 10% (vol/vol) ethanol at pH 3.0 stimulated killing by more than 3 log units in just 5 min, addition of 1.25% (vol/vol) ethanol resulted in less than 1 log unit of killing in 10 min. The ability of 5% (vol/vol) ethanol to stimulate killing at low pH and at elevated osmolarity was also dependent on the amplitude of the imposed stress, and an increase in the pH from 3.0 to 4.0 or a decrease in the sodium chloride concentration from 25 to 2.5% led to a marked reduction in the effectiveness of 5% (vol/vol) ethanol as an augmentative agent. Combinations of organic acids, low pH, and ethanol proved to be particularly effective bactericidal treatments; the most potent combination was pH 3.0, 50 mM formate, and 5 % (vol/vol) ethanol, which resulted in 5 log units of killing in just 4 min. Ethanol-enhanced killing correlated with damage to the bacterial cytoplasmic membrane. 相似文献
13.
Antivenoms are manufactured by the fractionation of animal plasma which may possibly be contaminated by infectious agents pathogenic to humans. This study was carried out to determine whether pre-existing antivenom production steps, as carried out by EgyVac in Egypt, may reduce viral risks. Two typical manufacturing steps were studied by performing down-scaled viral inactivation experiments: (a) a pH 3.3 pepsin digestion of diluted plasma at 30 degrees C for 1h, and (b) a caprylic acid treatment of a purified F(ab')2 fragment fraction at 18 degrees C for 1h. Three lipid-enveloped (LE) viruses [bovine viral diarrhoea virus (BVDV), pseudorabies virus (PRV), and vesicular stomatitis virus (VSV)] and one non-lipid-enveloped (NLE) virus [encephalomyocarditis virus (EMC)] were used as models. Kinetics of inactivation was determined by taking samples at 3 time-points during the treatments. The pH 3.3 pepsin digestion resulted in complete clearance of PRV (>7.0 log(10)) and in almost complete reduction of VSV (>4.5 but < or =6.4 log(10)), and in a limited inactivation of BVDV (1.7 log(10)). EMC inactivation was > or =2.5 but < or =5.7 log(10). The caprylic acid treatment resulted in complete inactivation of the 3 LE viruses tested: BVDV (>6.6 log(10)), PRV (>6.6 log(10)), and VSV (>7.0 log(10)). For EMC no significant reduction was obtained (0.7 log(10)). Cumulative reduction was >13.6, >11.5, >8.3 and > or =2.5 for PRV, VSV, BVDV and EMC, respectively. Therefore the current manufacturing processes of at least some animal antisera already include production steps that can ensure robust viral inactivation of LE viruses and moderate inactivation of a NLE virus. 相似文献
14.
The phytoplankton productivity of an acidic lake 总被引:1,自引:1,他引:0
The phytoplankton productivity over two years in a lake heavily loaded by acid mine drainage was very low. Algal assays indicated that below pH 5.5 the water, if buffered and fertilized with phosphorus, resulted in log growth. Above pH 5.5 algal log growth could be induced with phosphorus addition only. However, an in situ bag experiment was done in the lake and immediate bloom conditions of the indigenous algae resulted from phosphorus addition only, despite pH values below 5. 相似文献
15.
The killing of Listeria monocytogenes following exposure to low pH, organic acids, and osmotic stress was enhanced by the addition of 5% (vol/vol) ethanol. At pH 3, for example, the presence of this agent stimulated killing by more than 3 log units in 40 min of exposure. The rate of cell death at pH 3.0 was dependent on the concentration of ethanol. Thus, while the presence 10% (vol/vol) ethanol at pH 3.0 stimulated killing by more than 3 log units in just 5 min, addition of 1.25% (vol/vol) ethanol resulted in less than 1 log unit of killing in 10 min. The ability of 5% (vol/vol) ethanol to stimulate killing at low pH and at elevated osmolarity was also dependent on the amplitude of the imposed stress, and an increase in the pH from 3.0 to 4.0 or a decrease in the sodium chloride concentration from 25 to 2.5% led to a marked reduction in the effectiveness of 5% (vol/vol) ethanol as an augmentative agent. Combinations of organic acids, low pH, and ethanol proved to be particularly effective bactericidal treatments; the most potent combination was pH 3.0, 50 mM formate, and 5 % (vol/vol) ethanol, which resulted in 5 log units of killing in just 4 min. Ethanol-enhanced killing correlated with damage to the bacterial cytoplasmic membrane. 相似文献
16.
《Comparative biochemistry and physiology. A, Comparative physiology》1979,62(1):207-212
- 1. The hemoglobin of the Amazonian catfishPseudodoras sp. was isolated and characterized; it comprises a single component.
- 2. The hemoglobin's subunit composition is similar to that of other teleost hemoglobins. The apparent native molecular weight as determined by gel filtration is 66,000. The apparent subunit molecular weight is 14,300 by sodium dodecyl sulfate electrophoresis. The hemoglobin does not polymerize after oxidation by potassium ferricyanide.
- 3. The hemoglobin lacks a Root effect. A small Bohr effect is evident in the phosphate-free hemoglobin:Δlog p1/2/ΔpH is no more than about −0.1 to −0.2 and increases toΔlog p1/2/ΔpH = −0.4 in the presence of 1 mM ATP. The cooperativity, as determined byn of the Hill equation, is low, varying from 0.8 to 1.7 between pH 6.1 and 8.6.
- 4. Thep1/2 values of stripped hemoglobin solutions are extremely low, less than 0.5 mm Hg at all pH values examined between pH 6.1 and 9.0. The high oxygen affinity is reflected primarily in the CO combination rate which resembles that found in myoglobins and isolated subunits of human hemoglobin.
- 5. Both the CO combination rate and the O2 dissociation rate determined by stopped-flow spectrophotometry are pH and phosphate sensitive. Between pH 6.2 and 8.1 the COon rate increases about 5-fold in the phosphate-free hemoglobin. Addition of 1 mM ATP causes a depression in the rate at all pH values examined. The O2off rate decreases 7-fold going from pH 6.0 to 8.2 in stripped hemoglobin solutions. Addition of 1 mM ATP induces a 10-fold decrease over the same range. At pH values below 6.0 a depression in the O2off rate occurs in the stripped hemoglobin, indicative of an acid Bohr effect.
17.
J L Bailly M Chambon H Peigue-Lafeuille H Laveran C De Champs D Beytout 《Applied and environmental microbiology》1991,57(4):1156-1160
The activity of glutaraldehyde (GTA) at low concentrations (less than 2%) against poliovirus was assessed by a suspension procedure. The inactivation kinetics showed that concentrations of less than or equal to 0.10% were effective against purified poliovirus at pH 7.2; a 1 log10 reduction was obtained in 70 min with 0.02% GTA, and a 3 log10 reduction was obtained in 30 min with 0.10% GTA. GTA activity at low concentrations was greatly enhanced at alkaline pH, but was completely abolished at acid pH. In contrast, the inactivation assays on poliovirus RNA showed that it was highly resistant to GTA at concentrations up to 1.0% at pH 7.2. At pH 8.3 a low inactivation was noticed with 1.0% GTA. Our results are of relevance to hospital practice in digestive endoscopy investigations because there has been an increasing tendency to use low concentrations of GTA and very short contact times in disinfection procedures. 相似文献
18.
Activity of glutaraldehyde at low concentrations (less than 2%) against poliovirus and its relevance to gastrointestinal endoscope disinfection procedures. 总被引:5,自引:3,他引:2
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J L Bailly M Chambon H Peigue-Lafeuille H Laveran C De Champs D Beytout 《Applied microbiology》1991,57(4):1156-1160
The activity of glutaraldehyde (GTA) at low concentrations (less than 2%) against poliovirus was assessed by a suspension procedure. The inactivation kinetics showed that concentrations of less than or equal to 0.10% were effective against purified poliovirus at pH 7.2; a 1 log10 reduction was obtained in 70 min with 0.02% GTA, and a 3 log10 reduction was obtained in 30 min with 0.10% GTA. GTA activity at low concentrations was greatly enhanced at alkaline pH, but was completely abolished at acid pH. In contrast, the inactivation assays on poliovirus RNA showed that it was highly resistant to GTA at concentrations up to 1.0% at pH 7.2. At pH 8.3 a low inactivation was noticed with 1.0% GTA. Our results are of relevance to hospital practice in digestive endoscopy investigations because there has been an increasing tendency to use low concentrations of GTA and very short contact times in disinfection procedures. 相似文献
19.
Clifford L. Slayman 《The Journal of general physiology》1965,49(1):69-92
Glass micropipette electrodes have been employed to study the transsurface potential difference of Neurospora crassa. For mature hyphae grown in agar cultures, the internal potential is large and negative, often exceeding -200 mv. The potential is sensitive to the concentrations of extracellular potassium, sodium, hydrogen, and calcium ions, but does not vary in a manner which is readily explained by ionic diffusion potentials. With extracellular solutions containing only potassium chloride (or sulfate) and sucrose, the internal potential shifts toward zero (becomes less negative) at 45 mv per tenfold increase of potassium, over the range 0.1 to 10 mM. A similar result has been found with sodium, though the slope is only 33 mv/log unit. Calcium (1 mM) diminishes the influence of potassium and sodium by 60 to 70 per cent. As potassium or sodium is raised above 20 mM, the slope of the internal potential increases sharply to 85 to 90 mv/log unit, both in the presence and absence of calcium. With increasing hydrogen ion concentration, too, the internal potential shifts toward zero; in this case the slope is about 12 mv/pH unit at pH 9 and rises smoothly to 33 mv/pH unit at pH 3. All these phenomena are probably properties of the plasma membrane. The polysaccharide cell wall contains few fixed negative charges, has a low transverse resistance, and supports very little potential difference when separated from the plasma membrane. 相似文献
20.
Lactic acid bacteria (LAB) with antimicrobial activity and high exopolysaccharide (EPS) production ability isolated from sourdough were studied for their fermentation characteristics as potential new starter cultures. The values of pH, titratable acidity, and viable cell counts were 4.06+/-0.009-4.50+/- 0.015, 0.787+/-0.020%-1.172+/-0.018%, and 8.78+/-0.08-8.98+/- 0.06 log CFU/ml, respectively. In order to select probiotics with a high survival rate in the gut, isolates were tested to assess resistance against the artificial gastric acid and bile juice. Viable LAB counts were significantly (p<0.05) affected by the acidity. At pH 2.0, the total declines in the initial bacterial counts were 4.52+/-0.07 log for S. thermophilus St-Body-1, >7.98+/-0.03 log for E. flavescens DU-10, >7.95+/-0.05 log for E. faecium DU-12, and 3.15+/- 0.06 log for L. amylovorus DU-21. Among the strains, L. amylovorus DU-21 was the only strain that had bile tolerance under simulated gastrointestinal conditions. In order to improve EPS production by L. amylovorus DU- 21, the influence of carbon source was studied. When glucose was used as a carbon source, EPS production dramatically increased to 17.19+/-0.28 g/l (p<0.05). The maximum cell growth (10.012+/-0.012 log CFU/ml) and EPS production (18.71+/-0.19 g/l) were achieved when 15 g/ l of glucose was employed as the carbon source. 相似文献