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101.
J. J. O'Leary G. Browne R. J. Landers M. Crowley I. Bailey Healy J. T. Street A. M. Pollock J. Murphy M. I. Johnson F. A. Lewis et al. 《The Histochemical journal》1994,26(4):337-346
Summary Conventional solution-phase polymerase chain reaction (PCR) and in situ PCR/PCR in situ hybridization are powerful tools for retrospective analysis of fixed paraffin wax-embedded material. Amplification failure using these techniques is now encountered in some centres using archival fixed tissues. Such ailures may not only be due to absent target DNA sequences in the tissues, but may be a direct effect of the type of fixative, fixation time and/or fixation temperature used. The type of nucleic acid extraction procedure applied will also influence amplification results. This is particularly true with in situ PCR/PCR in situ hybridization.To examine these effects in solution-phase PCR, -globin gene was amplified in 100 mg pieces of tonsillar tissue fixed in Formal saline, 10% formalin, neutral buffered formaldehyde, Carnoy's, Bouin's, buffered formaldehyde sublimate, Zenker's, Helly's and glutaraldehyde at 0 to 4°C, room temperature and 37°C fixation temperatures and for fixation periods of 6, 24, 48 and 72 hours and 1 week. DNA extraction procedures used were simple boiling and 5 days' proteinase K digestion at 37°C. Amplified product was visible primarily yet variably from tissue fixed in neutral buffered formaldehyde and Carnoy's, whereas fixation in mercuric chloride-based fixatives produced consistently negative results. Room temperature and 37°C fixation temperature appeared most conducive to yielding amplifiable DNA template. Fixation times of 24 and 48 hours in neutral buffered formaldehyde and Carnoy's again favoured amplification.Fixed SiHa cells (containing 1–2 copies of HPV 16) were examined using PCR in situ hybridization for the amplification of HPV 16. Discrete and diffuse amplification signals were obtained. Neutral buffered formaldehyde fixation for 12–24 hours yielded amplifiable material suitable for use with PCR in situ hybridization. Overall amplification success within cellular preparations was 40%, with non-specific background staining also seen. Possible technical problems encountered with PCR in situ hybridization are discussed. 相似文献
102.
Pradip Bhatnagar William Huffman Andrew King Louis Pelus William Potts et al. 《Journal of Ornithology》1994,135(1):61-71
Research Notes on Avian Biology 1994: Selected Contributions from the 21st International Ornithological CongressMorphology and Physiology: Endocrinology
Subject: Endocrinology and chronobiology 相似文献103.
104.
Incomplete protection, but suppression of virus burden, elicited by subunit simian immunodeficiency virus vaccines. 总被引:12,自引:10,他引:2
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Z R Israel P F Edmonson D H Maul S P O'Neil S P Mossman C Thiriart L Fabry O Van Opstal C Bruck F Bex et al. 《Journal of virology》1994,68(3):1843-1853
We compared the efficacy of immunization with either simian immunodeficiency virus (SIV) Env glycoprotein (Env), Env plus Gag proteins (Gag-Env), or whole inactivated virus (WIV), with or without recombinant live vaccinia vector (VV) priming, in protecting 23 rhesus macaques (six vaccine and two control groups) from challenge with SIVmac251 clone BK28. Vaccination elicited high titers of syncytium-inhibiting and anti-Env (gp120/gp160) antibodies in all vaccinated macaques and anti-Gag (p27) antibodies in groups immunized with WIV or Gag-Env. Only WIV-immunized macaques developed anticell (HuT78) antibodies. After homologous low-dose intravenous virus challenge, we used frequency of virus isolation, provirus burden, and change in antibody titers to define four levels of resistance to SIV infection as follows. (i) No infection ("sterilizing" immunity) was induced only in WIV-immunized animals. (ii) Abortive infection (strong immunity) was defined when virus or provirus were detected early in the postchallenge period but not thereafter and no evidence of virus or provirus was detected in terminal tissues. This response was observed in two animals (one VV-Env and one Gag-Env). (iii) Suppression of infection (incomplete or partial immunity) described a gradient of virus suppression manifested by termination of viremia, declining postchallenge antibody titers, and low levels (composite mean = 9.1 copies per 10(6) cells) of provirus detectable in peripheral blood mononuclear cells or lymphoid tissues at termination (40 weeks postchallenge). This response occurred in the majority (8 of 12) of subunit-vaccinated animals. (iv) Active infection (no immunity) was characterized by persistent virus isolation from blood mononuclear cells, increasing viral antibody titers postchallenge, and high levels (composite mean = 198 copies per 10(6) cells) of provirus in terminal tissues and blood. Active infection developed in all controls and two of three VV-Gag-Env-immunized animals. The results of this study restate the protective effect of inactivated whole virus vaccines produced in heterologous cells but more importantly demonstrate that a gradient of suppression of challenge virus growth, reflecting partial resistance to SIV infection, is induced by subunit vaccination. The latter finding may be pertinent to studies with human immunodeficiency virus vaccines, in which it is plausible that vaccination may elicit significant suppression of virus infection and pathogenicity rather than sterilizing immunity. 相似文献
105.
106.
Probing protein-cofactor interactions in the terminal oxidases by second derivative spectroscopy: study of bacterial enzymes with cofactor substitutions and heme A model compounds.
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J. S. Felsch M. P. Horvath S. Gursky M. R. Hobaugh P. N. Goudreau J. A. Fee W. T. Morgan S. J. Admiraal M. Ikeda-Saito T. Fujiwara et al. 《Protein science : a publication of the Protein Society》1994,3(11):2097-2103
Second derivative absorption spectra are reported for the aa3-cytochrome c oxidase from bovine cardiac mitochondria, the aa3-600 ubiquinol oxidase from Bacillus subtilis, the ba3-cytochrome c oxidase from Thermus thermophilis, and the aco-cytochrome c oxidase from Bacillus YN-2000. Together these enzymes provide a range of cofactor combinations that allow us to unequivocally identify the origin of the 450-nm absorption band of the terminal oxidases as the 6-coordinate low-spin heme, cytochrome a. The spectrum of the aco-cytochrome c oxidase further establishes that the split Soret band of cytochrome a, with features at 443 and 450 nm, is common to all forms of the enzyme containing ferrocytochrome a and does not depend on ligand occupancy at the other heme cofactor as previously suggested. To test the universality of this Soret band splitting for 6-coordinate low-spin heme A systems, we have reconstituted purified heme A with the apo forms of the heme binding proteins, hemopexin, histidine-proline-rich glycoprotein and the H64V/V68H double mutant of human myoglobin. All 3 proteins bound the heme A as a (bis)histidine complex, as judged by optical and resonance Raman spectroscopy. In the ferroheme A forms, none of these proteins displayed evidence of Soret band splitting. Heme A-(bis)imidazole in aqueous detergent solution likewise failed to display Soret band splitting. When the cyanide-inhibited mixed-valence form of the bovine enzyme was partially denatured by chemical or thermal means, the split Soret transition of cytochrome a collapsed into a single band at 443 nm.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
107.
本研究从日本落叶松引种区林地棕壤9个主要理化性状及与之配套的17株优势木材料入手,采用多变量分析方法筛选出影响该地区日本落叶松生长的主要肥力因子为氮及磷,並据此将该林地棕壤划分为9个肥力类型。在进行聚类分析时,本研究的创新在于以任意二样本点在各主成份上的坐标值为变量,以各主成份的贡献率为权重,重新定义欧氏距离公式,对主成份坐标值聚类,使其更为合理、直观。同时采用回归分析拟合出理化性状间的多个回归方程。此研究为该林地科学施肥、速生丰产、永续利用提供了可靠的数量依据。 相似文献
108.
将54例冠心病、高血压病、糖尿病及脑梗塞病人,随机分为对照组与清栓酶组。对照组26例,常规药物治疗,清栓酶组28例,在常规药物治疗基础上加用蝮蛇清栓酶0.5u,稀释静脉点滴,一日一次,14天为一疗程。观察到蝮蛇清栓酶能明显升高超氧化物岐化酶活性(P<0.01),明显降低过氧化脂质的终末代谢产物丙二醛含量(P<0.01),说明蝮蛇清栓酶能有效减轻自由基损伤。 相似文献
109.
HBIG,无环鸟苷,干扰素联合对慢性乙型肝炎抗病毒效应观察 总被引:1,自引:0,他引:1
本文报道血清HBV复制标志阳性的慢乙肝54例,随机分为治疗组及对照组各27例进行HBIG、无环鸟苷、干扰素联合近、远期抗病毒效应观察。治疗组为无环鸟苷第一周按25~20mg/kg/d计后改17~15mg/kg/d×53天,共60天;人白细胞干扰素1×106U肌注每周3次×4周,后改1.0×106U肌注每周2次×6周,共10周;HBIG400U肌注隔日1次,共10周,对照组仅给予一般“保肝”药物。其中治疗组18例,对照组19例进行治后半年到2年追踪观察,结果近、远期HBcAg、DNAP、HBV-DNA阴转率治疗组均高于对照组,其中治疗组近、远期HBcAg,HBV-DNA阴转率均达40%以上,明显高于对照组(P<0.05~0.01),治疗组近、远期各有4例及2例HBsAg阴转,而对照组则无一例阴转,从近、远期综合抗病毒效应观察,治疗组全阴率分别为33.3%、44.4%,而对照组分别为3.79%及0%,P<0.01,治疗组无明显毒副反应。对比单用无环鸟苷,全阴率31.8%;无环鸟苷加干扰素两药联合全阴率37.5%,均有所提高,达到44.4%,值得进一步研究。 相似文献
110.
应用底物膜技术检测130例正常精液,精子顶体酶活性百分率的正常值下限为57%。459例不孕症病人精液分析,无精症25例,其余434例中75%精子顶体酶活性正常。实验表明精子密度对数值与顶体酶活性百分率之间有正相关,r=0.84(P<0.01),回归方程为顶体酶活性百分率y=48.43%+(8.9%)(log精子计数)。活动精子百分率与顶体酶活性之间有密切正相关,r=0.967,(P<0.01),顶体酶活性y=38.6%+0、36x%。前向活跃直线运动精子百分率与顶体酶活性之间也有密切相关.r=0.96,(P<0.01),顶体酶活性y=34.21%+0.61x%。 相似文献