Biochemical and molecular medicine最新文献_第2页

Genomic DNA from Mice: A Comparison of Recovery Methods and Tissue Sources 小鼠基因组DNA:恢复方法和组织来源的比较

Biochemical and molecular medicine Pub Date : 1997-12-01 DOI: 10.1006/bmme.1997.2637

John R. Hofstetter , Aiwu Zhang , Aimee R. Mayeda , Tim Guscar , John I. Nurnberger Jr. , Debomoy K. Lahiri

{"title":"Genomic DNA from Mice: A Comparison of Recovery Methods and Tissue Sources","authors":"John R. Hofstetter , Aiwu Zhang , Aimee R. Mayeda , Tim Guscar , John I. Nurnberger Jr. , Debomoy K. Lahiri","doi":"10.1006/bmme.1997.2637","DOIUrl":"10.1006/bmme.1997.2637","url":null,"abstract":"<div><p>Our aim is to identify an extraction method and the source of mouse tissue(s) that could allow a high-resolution genomic scan from a living mouse. We compared and optimized two methods for yield, purity of DNA, and their use in the polymerase chain reaction (PCR) of DNA extracted from different mouse tissues. In addition to whole blood, tissue samples from the brain, liver, testis, and tail were included in this study. The Rapid Method (RM) is preferable for the whole blood samples and testis and brain tissue samples because it is quicker, less toxic, and more cost-effective than the proteinase K method (PM). For liver the PM produced higher yields of DNA with less degradation than the RM. For tail tip, the PM produced a higher yield of DNA, but the RM resulted in a higher yield of PCR product. From a living mouse, a tail snip generated a sufficient amount of DNA for several hundred PCRs but not a complete genomic scan. We suggest that the RM can be used to extract genomic DNA for a complete genomic scan which requires either testicular tissues or repeated blood samples from the suborbital sinus over several months without sacrificing the animal.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"62 2","pages":"Pages 197-202"},"PeriodicalIF":0.0,"publicationDate":"1997-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2637","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20371189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 55

Tetrahydrobiopterin in the Treatment of Infantile Hypertrophic Pyloric Stenosis 四氢生物蝶呤治疗婴儿肥厚性幽门狭窄

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2628

Christian P. Braegger , Marcus Schwöbel , Jakob von Känel , Ernst R. Werner , Beat Thöny , Nenad Blau

引用次数: 6

Therapeutic Potential and Mechanism of Action of Oligonucleotides and Ribozymes 寡核苷酸和核酶的治疗潜力及其作用机制

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2631

Yan Lavrovsky, Shuo Chen, Arun K. Roy

引用次数: 23

Cholesteryl Ester Storage Disease: Relationship between Molecular Defects andin SituActivity of Lysosomal Acid Lipase 胆固醇酯贮藏病:溶酶体酸性脂肪酶分子缺陷与原位活性的关系

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2626

Isabelle Redonnet-Vernhet, Martine Chatelut, Jean-Pierre Basile, Robert Salvayre, Thierry Levade

{"title":"Cholesteryl Ester Storage Disease: Relationship between Molecular Defects andin SituActivity of Lysosomal Acid Lipase","authors":"Isabelle Redonnet-Vernhet, Martine Chatelut, Jean-Pierre Basile, Robert Salvayre, Thierry Levade","doi":"10.1006/bmme.1997.2626","DOIUrl":"10.1006/bmme.1997.2626","url":null,"abstract":"<div><p>The molecular defects in the<em>LIPA</em>gene encoding the lysosomal acid lipase (LAL) were investigated in two unrelated patients affected with cholesteryl ester storage disease (CESD), an autosomal recessive disorder associated with LAL-deficient activity. In cell lysates from both patients there was a severely reduced LAL activity. In a female patient, nucleotide sequencing of amplified LAL genomic DNA or reverse-transcribed mRNA demonstrated that she was a compound heterozygote for two previously reported mutations, a G → A transition at position −1 of the exon 8 splice donor site, resulting in skipping of the complete exon 8, and a C<sup>923</sup>→ T substitution leading to the replacement of His<sup>274</sup>to Tyr. The second, male CESD patient was heterozygous for the splice junction mutation and a yet undescribed C → T substitution at position 233, which introduces a premature in-frame termination codon. The functional consequences of these genetic alterations were evaluated for the first time by studying the catabolic turnover of radiolabeled cholesteryl oleate in intact cells. A lower<em>in situ</em>residual LAL activity was found in cells carrying the stop codon mutation than in cells having the His<sup>274</sup>→ Tyr substitution. Since the severely reduced LAL activity was seen in cells from an adult patient with a mild CESD, we conclude that there is no simple direct correlation between the LAL molecular lesions and the biochemical and clinical phenotypes.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"62 1","pages":"Pages 42-49"},"PeriodicalIF":0.0,"publicationDate":"1997-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2626","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20298361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 25

Enhanced Resistance of Adriamycin-Treated MCR-5 Lung Fibroblasts by Increased Intracellular Glutathione Peroxidase and Extracellular Antioxidants 通过增加细胞内谷胱甘肽过氧化物酶和细胞外抗氧化剂增强阿霉素处理的MCR-5肺成纤维细胞的耐药性

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2612

Angelo Vanella , Agata Campisi, Claudia di Giacomo, Valeria Sorrenti, Giuseppe Vanella, Rosaria Acquaviva

引用次数: 10

Sulfation of Chondroitin/Dermatan Sulfate by Cystic Fibrosis Pancreatic Duct Cells Is Not Different from Control Cells 囊性纤维化胰腺管细胞对软骨素/硫酸皮肤素的磺化作用与对照细胞没有区别

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2625

Warren G. Hill , Gregory S. Harper , Tina Rozaklis , John J. Hopwood

{"title":"Sulfation of Chondroitin/Dermatan Sulfate by Cystic Fibrosis Pancreatic Duct Cells Is Not Different from Control Cells","authors":"Warren G. Hill , Gregory S. Harper , Tina Rozaklis , John J. Hopwood","doi":"10.1006/bmme.1997.2625","DOIUrl":"10.1006/bmme.1997.2625","url":null,"abstract":"<div><p>Cystic fibrosis is associated with mutations of the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-regulated plasma membrane chloride channel. Cystic fibrosis patients have been reported to possess elevated sulfation of glycoconjugates, which may contribute to the pathogenesis of the disease. Sulfation of glycosaminoglycans by a cystic fibrosis pancreatic adenocarcinoma cell line homozygous for ΔF<sub>508</sub>(CFPAC-1), a control pancreatic cell line (PANC-1), two CFPAC-1 cell lines transfected with the gene for CFTR (PLJ-CFTR-4.7, TR20), and a mock-transfected CFPAC-1 control (PLJ-6) was investigated. Cells were radiolabeled with [<sup>35</sup>S]sulfate and [<sup>3</sup>H]glucosamine, and glycosaminoglycans secreted into the medium after 24 and 72 h were isolated. Chondroitinase ABC digestion of chondroitin/dermatan sulfate allowed the recovery of disaccharides which were analyzed for their degree of sulfation by strong anion-exchange HPLC. No differences in the extent of sulfation by any of the cell lines were noted. However, glycoaminoglycans synthesized by cystic fibrosis cells consistently exhibited twofold higher [<sup>35</sup>S]-sulfate:[<sup>3</sup>H]glucosamine ratios than the controls. We conclude that CFTR plays no role in the sulfation of chondroitin/dermatan sulfate by pancreatic cells and that isotope incorporation ratios alone are insufficient evidence of changes in sulfation levels.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"62 1","pages":"Pages 85-94"},"PeriodicalIF":0.0,"publicationDate":"1997-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2625","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20300188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

A Common 844INS68 Insertion Variant in the Cystathionine β-Synthase Gene 半胱硫氨酸β-合成酶基因中常见的844INS68插入变异

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2623

Leo A.J. Kluijtmans , Godfried H.J. Boers , Frans J.M. Trijbels , Henriëtte M.A. van Lith-Zanders , Lambert P.W.J. van den Heuvel , Henk J. Blom

引用次数: 70

The Pathogenetic Role of Heme in Pregnancy-Induced Hypertension-like Disease in Ewes 血红素在母羊妊娠性高血压样疾病中的致病作用

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2602

Gyula Tálosi , Ilona Németh , Erzsébet Nagy , Sándor Pintér

{"title":"The Pathogenetic Role of Heme in Pregnancy-Induced Hypertension-like Disease in Ewes","authors":"Gyula Tálosi , Ilona Németh , Erzsébet Nagy , Sándor Pintér","doi":"10.1006/bmme.1997.2602","DOIUrl":"10.1006/bmme.1997.2602","url":null,"abstract":"<div><p>Toxicosis syndrome of fasting pregnant ewes has a close similarity to human preeclampsia (hypertension, albuminuria). The common etiological factor might be oxidative hemolysis and heme-induced endothelial damage. Ewes (5 starving, 5 control) at 130–135 gestational days with a 96-h fasting period followed by refeeding were used. Blood pressure, platelet count, electrolytes, kidney and liver function parameters, as well as plasma glucose, hemoglobin/heme, free thiol groups and Trolox equivalent antioxidant capacity, and plasma iron and ferritin levels were measured. Statistical significance was assessed using Student's<em>t</em>test (<em>P</em>< 0.05). Besides hypertension and renal disturbances, hemolysis, elevated liver enzymes and low platelet count, characteristic of human HELLP syndrome, were also present. In the first 24 h of glucose deprivation there was a significant rise in both the plasma hemoglobin/heme and indirect bilirubin concentrations. The antioxidant free thiol levels decreased significantly the next day, without any change in the total antioxidant capacity of the plasma. While the loss of calcium and magnesium levels related to the similarity to preeclampsia, reduced plasma iron concentrations referred to species differences in iron homeostasis. An oxidative stress causing hemolysis in a glucose-6-phosphate dehydrogenase-deficient animal model was proven by the loss of free thiols after glucose deprivation. The activation of the oxidative stress protein heme oxygenase was a signal of endothelial cell injury, the primary cause of pregnancy-induced hypertension.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"62 1","pages":"Pages 58-64"},"PeriodicalIF":0.0,"publicationDate":"1997-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2602","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20298363","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 13

Oxidation of Bilirubin by Rat Brain Mitochondrial Membranes—Genetic Variability 大鼠脑线粒体膜氧化胆红素的遗传变异

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2618

Thor Willy Ruud Hansen , Steve Tommarello, Jeffrey W. Allen

{"title":"Oxidation of Bilirubin by Rat Brain Mitochondrial Membranes—Genetic Variability","authors":"Thor Willy Ruud Hansen , Steve Tommarello, Jeffrey W. Allen","doi":"10.1006/bmme.1997.2618","DOIUrl":"10.1006/bmme.1997.2618","url":null,"abstract":"<div><p>Bilirubin is oxidized by brain mitochondrial membranes at a rate which may contribute significantly to clearance of bilirubin from brain. Different strains of congenitally jaundiced rats (Gunn rats) vary widely as far as the mortality rate of the homozygous (jaundiced) pups. Because the ability to oxidize bilirubin in brain may protect against toxicity, we hypothesized that the ability to oxidize bilirubin would be lower in Gunn rat strains (ACI/N-j) with a high mortality rate in the homozygous pups. Mitochondria were obtained from young rat brains by differential centrifugation in sucrose gradients. The mitochondria were ruptured by sonication. The change in optical density of a bilirubin solution at 440 nm was measured over time following addition of the membrane suspension. The rate of bilirubin oxidation was significantly lower in rats of the RHA/N-j strain both at 7–8 days of age and in adults, compared to rats of the ACI/N-j and the Sprague-Dawley strains at the same age points. Differences in mortality rates between the RHA/N-j and the ACI/N-j strains of Gunn rats could not be explained on the basis of differences in the ability of brain mitochondrial membranes to oxidize bilirubin, as these activities were lower in the RHA/N-j rats, which also have lower mortality rates, but higher in the ACI/N-j rats, which have remarkably high mortality rates. This study also confirmed previous findings relative to age maturation of the enzyme activity.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"62 1","pages":"Pages 128-131"},"PeriodicalIF":0.0,"publicationDate":"1997-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2618","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20300194","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 15

The Old and the New in p53 Functional Regulation p53功能调控的新与旧

Biochemical and molecular medicine Pub Date : 1997-10-01 DOI: 10.1006/bmme.1997.2616

Lucia Magnelli, Marco Ruggiero, Vincenzo Chiarugi

引用次数: 10