Biochemical and molecular medicine最新文献

{"title":"Expression of PAX3 in Ewing's Sarcoma Family of Tumors","authors":"Theodor W. Schulte ,&nbsp;Jeffrey A. Toretsky ,&nbsp;Elisabeth Ress ,&nbsp;Lee Helman ,&nbsp;Leonard M. Neckers","doi":"10.1006/bmme.1997.2567","DOIUrl":"10.1006/bmme.1997.2567","url":null,"abstract":"<div><p>The Ewing's sarcoma family of tumors (ESFT) is the second most common pediatric malignancy originating in the bone and is characterized by the t(11; 22) translocation. PAX3, a member of the paired box family of genes, is expressed during embryonal development of neural crest cells and is involved in the t(2; 13) translocation found in alveolar rhabdomyosarcoma. Since ESFTs are believed to be derived from neural crest tissue, we screened a series of Ewing's sarcoma and peripheral neuroectodermal tumor cell lines and tumor specimens for expression of PAX3. We found expression of PAX3 in most, but not all, of the specimens analyzed, including cell lines and patient material.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Pages 121-126"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2567","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20116391","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}

{"title":"Oxidation of Bilirubin by Brain Mitochondrial Membranes—Dependence on Cell Type and Postnatal Age","authors":"Thor Willy Ruud Hansen ,&nbsp;Jeffrey W. Allen","doi":"10.1006/bmme.1996.2565","DOIUrl":"10.1006/bmme.1996.2565","url":null,"abstract":"<div><p>Bilirubin is oxidized by brain mitochondrial membranes at a rate which may contribute significantly to clearance of bilirubin from the brain. Neurons appear to be more sensitive to bilirubin toxicity than glial cells. Clinical experience has suggested that sensitivity to bilirubin neurotoxicity may be greater in the neonate than later in life. We hypothesized that the ability to oxidize bilirubin would be lower in mitochondrial membranes from a pure neuronal compared to a mixed glial/neuronal source, and lower in immature than more mature brains. Mitochondria of synaptosomal and nonsynaptosomal origin were obtained from young rat brains by differential centrifugation in sucrose gradients. Synaptosomes were lysed by hypoosmotic treatment, and 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 higher in nonsynaptic than in synaptic mitochondrial membranes [99.1 ± 42.3 (mean ± SD) vs 69.9 ± 30.9 pmol/min/mg protein,<em>t</em>= 4.835,<em>P</em>= 0.0003]. “Crude” mitochondrial membranes were obtained by differential centrifugation in sucrose from the forebrains of rats of 7, 14, and 21 days postnatal age as well as adults, and from rabbits of 1 and 7 days postnatal age as well as adults. In both species the rates of bilirubin oxidation increased significantly with postnatal age (rats:<em>F</em>= 55.3,<em>P</em>&lt; 0.0001; rabbits:<em>F</em>= 101,<em>P</em>&lt; 0.0001). Mitochondrial membranes from a pure neuronal source oxidize bilirubin at a significantly lower rate than membranes from a mixed glial/neuronal source. This suggests that neurons may be less able to detoxify bilirubin locally and thus might contribute to the apparent higher sensitivity to bilirubin toxicity in these cells vs glia. Similarly, the lower bilirubin-oxidizing ability of mitochondrial membranes from immature brains seems compatible with the clinical impression of increased vulnerability to bilirubin neurotoxicity in the newborn.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Pages 155-160"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1996.2565","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20116292","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}

{"title":"Insulin Resistance and the Transcription of the Glucose-6-Phosphatase Gene in Newborn Dogs","authors":"Bing-cheng Feng,&nbsp;Jixuan Li,&nbsp;Robert M. Kliegman","doi":"10.1006/bmme.1996.2563","DOIUrl":"10.1006/bmme.1996.2563","url":null,"abstract":"<div><p>In the present report changes in the mRNA level of glucose-6-phosphatase (G6Pase; EC 3.1.39) in newborn and adult dogs<em>in vivo</em>were studied to further test the hypotheses that neonatal hyperglycemia may be due to unsuppressed gluconeogenesis by insulin and that the antidiabetic role of insulin-like growth factor-1 (IGF-1) may be intact in newborn dogs who have consistently demonstrated insulin resistance. Our results were the following: (i) Both renal and hepatic G6Pase mRNA were expressed at birth and increased with time during a 24-h period of fasting after birth. (ii) The renal G6Pase mRNA levels in newborn dogs did not respond to either insulin or epinephrine. (iii) Hyperinsulinemia lowered the liver G6Pase mRNA by only 16.3% in newborn dogs, but reduced the liver G6Pase mRNA to an undetectable level in adult dogs. (iv) Hyperglycemia decreased the hepatic G6Pase mRNA by 14.3% in newborn dogs under hyperinsulinemia. (v) Infused epinephrine did not elevate the hepatic G6Pase mRNA level in newborn dogs in the presence of hyperglycemia and hyperinsulinemia. (vi) In newborn dogs, hyper-IGF-1 rapidly reduced the hepatic G6Pase mRNA level by 50%, and hypoglycemia was unable to elevate the hepatic G6Pase mRNA level under the hyper-IGF-1. We concluded that the reduced rate of suppression of transcription of the liver G6Pase gene by insulin in newborn dogs may reflect the unsuppressed neonatal hepatic gluconeogenesis due to insulin resistance and that the physiological roles of IGF-1 seemed to be intact in newborn dogs and may be not responsible for neonatal hyperglycemia.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Pages 134-141"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1996.2563","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20116393","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}

{"title":"Author Index for Volume 60","authors":"","doi":"10.1006/bmme.1997.2587","DOIUrl":"https://doi.org/10.1006/bmme.1997.2587","url":null,"abstract":"","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Page 194"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2587","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136895285","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}

{"title":"Identification of a Ferritin Light Chain Pseudogene Near the Glycerol Kinase Locus in Xp21 by cDNA Amplification for Identification of Genomic Expressed Sequences","authors":"Weiwen Guo ,&nbsp;Volker Adams ,&nbsp;Jestina Mason ,&nbsp;Edward R.B. McCabe","doi":"10.1006/bmme.1996.2566","DOIUrl":"10.1006/bmme.1996.2566","url":null,"abstract":"<div><p>We used cDNA amplification for identification of genomic expressed sequences (CAIGES) to identify genes in the glycerol kinase region of the human X chromosome. During these investigations we identified the sequence for a ferritin light chain (FTL) pseudogene in this portion of Xp21. A human liver cDNA library was amplified by vector primers, labeled, and hybridized to Southern blots of<em>Eco</em>RIdigested human genomic DNA from cosmids isolated from yeast artificial chromosomes in the glycerol kinase region of Xp21. A 3.1-kb restriction fragment hybridized with the cDNA library, was subcloned and sequenced, and a 440-bp intronless sequence was found with strong similarity to the FTL coding sequence. Therefore, the FTL pseudogene that had been mapped previously to Xp22.3–21.2 was localized specifically to the glycerol kinase region. The CAIGES method permits rapid screening of genomic material and will identify genomic sequences with similarities to genes expressed in the cDNA library used to probe the cloned genomic DNA, including pseudogenes.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Pages 169-173"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1996.2566","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20116294","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}

{"title":"Quantitative–Competitive Polymerase Chain Reaction for Rapid Susceptibility Testing ofMycobacterium tuberculosisto Isoniazid","authors":"Benoosh Afghani,&nbsp;Harris R. Stutman","doi":"10.1006/bmme.1997.2573","DOIUrl":"10.1006/bmme.1997.2573","url":null,"abstract":"<div><p>The objective of this study was to determine whether quantitative–competitive polymerase chain reaction (QC-PCR) can be used for rapid susceptibility testing of<em>Mycobacterium tuberculosis</em>(MTB). QC-PCR was used to determine relative amounts of mycobacterial DNA inoculated at different isoniazid (INH) concentrations. A total of six different INH-sensitive (INH-S) and five INH-resistant (INH-R) strains were inoculated in the presence of 0.0, 0.2, 1.0, and 10.0 μg/ml of INH. DNA was quantified using QC-PCR at Week 0 and weekly thereafter for 3 weeks. For the QC-PCR, 10-fold dilutions of control (240 bp) DNA having the same primer set as the target DNA (123 bp) were used. The amount of target DNA was estimated by using known amounts of the internal standard. For INH-S isolates there was ≥1 log difference in DNA concentration in the presence of each INH concentration compared to that of the control within 1 to 3 weeks. In contrast, for INH-R isolates there were no apparent differences in DNA concentration between the control suspensions and those containing 0.2 and 1.0 μg/ml INH during the 3-week incubation period. The highest INH concentration (10 μg/ml), however, did abolish the DNA increase seen in the other MTB suspensions. This preliminary study suggests that by using concentrations of 0.2 or 1.0 μg/ml of INH, QC-PCR may differentiate INH-R and INH-S MTB isolates within 1 week. This method may be of particular value when applied directly to clinical specimens with varying numbers of bacilli.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Pages 182-186"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2573","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20116296","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}

{"title":"Detection of Epstein–Barr Viral DNA in Serum Using Rapid-Cycle PCR","authors":"C.Lars Mouritsen ,&nbsp;Carl T. Wittwer ,&nbsp;Gudrun Reed ,&nbsp;Taiyaba M. Khan ,&nbsp;Thomas B. Martins ,&nbsp;Troy D. Jaskowski ,&nbsp;Christine M. Litwin ,&nbsp;Harry R. Hill","doi":"10.1006/bmme.1997.2571","DOIUrl":"10.1006/bmme.1997.2571","url":null,"abstract":"<div><p>Our study describes the comparison of a rapid nested PCR assay to standard serology techniques for the detection of Epstein–Barr virus (EBV) in serum. The sera of 81 patients with suspected EBV infection were analyzed; 54 were positive for one or more of the standard serology markers, i.e., IgM viral capsid antigen (VCA), IgG-VCA, Epstein–Barr nuclear antigen 1 (EBNA-1), and early antigen (EA), and 27 were negative for all serology markers. The sera from 15 normal healthy blood donors were also included. No EBV DNA was detected in any of the 15 blood donor samples or in any of the 27 samples with negative serology results. Eleven samples (20%) of the 54 with positive EBV serology results were positive for EBV DNA. Of these samples, 9 were EBV IgM-VCA positive and anti-EBNA negative, suggesting acute infection. One of the 11 samples had high titers of IgM-VCA, IgG-VCA, anti-EBNA, and anti-EA. The last of the 11 samples was from a patient with acute infectious mononucleosis without sufficient sample volume for EBV serology testing. Seventeen of the total 96 samples from the study were IgM-VCA positive and anti-EBNA negative and 9 of these 17 samples (53%) tested positive for EBV DNA. These data suggest that the detection of EBV DNA by PCR in serum may be a useful indicator of active infection rather than latent virus.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Pages 161-168"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2571","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20116293","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}

{"title":"Oral Ingestion of Mannose Elevates Blood Mannose Levels: A First Step toward a Potential Therapy for Carbohydrate-Deficient Glycoprotein Syndrome Type I","authors":"Gordon Alton ,&nbsp;Susanne Kjaergaard ,&nbsp;James R. Etchison ,&nbsp;Flemming Skovby ,&nbsp;Hudson H. Freeze","doi":"10.1006/bmme.1997.2574","DOIUrl":"10.1006/bmme.1997.2574","url":null,"abstract":"<div><p>Carbohydrate-deficient glycoprotein syndrome type I (CDGS) is an inherited metabolic disorder with multisystemic abnormalities resulting from a failure to add entire N-linked oligosaccharide chains to many glycoproteins. Fibroblasts from these patients also abnormally glycosylate proteins, but this lesion is corrected by providing 250 μm mannose to the culture medium. This correction of protein glycosylation suggests that providing dietary mannose to elevate blood mannose concentrations might also remedy some of the underglycosylation observed in these patients. We find that ingested mannose is efficiently absorbed and increases blood mannose levels in both normal subjects and CDGS patients. Blood mannose levels increased in a dose-dependent fashion with increasing oral doses of mannose (0.07–0.21 g mannose/kg body weight). Peak blood mannose concentrations occurred at 1–2 h following ingestion and the clearance half-time was approximately 4 h. Doses of 0.1 g mannose/kg body weight given at 3-h intervals maintained blood mannose concentrations at levels 3- to 5-fold higher than the basal level in both normal controls (∼55 μm) and CDGS patients. No side effects were observed for this dosage regimen. These results establish the feasibility of using mannose as a potential therapeutic dietary supplement (nutraceutical) to treat CDGS patients.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Pages 127-133"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2574","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20116392","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}

{"title":"The Relationship of Genotype to Phenotype in Phenylalanine Hydroxylase Deficiency1","authors":"Richard Koch ,&nbsp;Karol Fishler ,&nbsp;Colleen Azen ,&nbsp;Per Guldberg ,&nbsp;Flemming Güttler","doi":"10.1006/bmme.1997.2577","DOIUrl":"10.1006/bmme.1997.2577","url":null,"abstract":"<div><p>Seventy-two adults with phenylketonuria were evaluated to investigate the genotypic relationship to phenotype. Patient data were collected by chart review and medical follow-up as well as current psychological evaluation. Nineteen diagnosed neonatally had remained on a phenylalanine-restricted diet all their lives, whereas 34 who were also diagnosed on newborn screening had discontinued dietary restriction during childhood. Nineteen others who were born prior to newborn screening were diagnosed later than the newborn period on clinical grounds but have remained on dietary restriction. Comparison between intellectual ability, academic achievement, and mental illness was made with degree of diet control as defined by range of blood phenylalanine levels over time. Diet discontinuation in childhood did not significantly lower IQ per se but appeared to diminish academic achievement. The lowest IQ scores were associated with poor dietary restriction of phenylalanine in the diet during childhood. While there appears to be a strong genotypic relationship to phenotypic metabolic parameters in phenylketonuria, there does not seem to be a similar relationship to intellectual ability in adults. Mutation R408W was not strongly related to the occurrence of mental illness in this sample. We conclude that dietary restriction of phenylalanine neonatally and good control contributed to normal intellectual development. Continuation of dietary treatment into adulthood appeared to improve academic achievement in patients with severe phenylalanine hydroxylase mutations.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 2","pages":"Pages 92-101"},"PeriodicalIF":0.0,"publicationDate":"1997-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1997.2577","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20116386","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}

{"title":"Functional Defects in Lysosomal Enzymes in Autosomal Dominant Polycystic Kidney Disease (ADPKD): Abnormalities in Synthesis, Molecular Processing, Polarity, and Secretion","authors":"Patricia A. Hartz,&nbsp;Patricia D. Wilson","doi":"10.1006/bmme.1996.2542","DOIUrl":"10.1006/bmme.1996.2542","url":null,"abstract":"<div><p>The phenotype of autosomal dominant polycystic kidney disease (ADPKD) is characterized by basement membrane abnormalities, hyperproliferation, and alterations in epithelial cell polarity. Since proteinases have been implicated in matrix degradation and growth factor activation, lysosomal enzymes were compared in normal and ADPKD tissues and cell cultures. Acidic proteolytic activity (azocasein) was reduced in ADPKD, and specific enzymatic assays detected disease-dependent decreases in the specific activities of β-galactosidase, β-hexosaminidase, and cathepsins, B, L, and H. Cathepsin D-specific activities were unchanged. Lucifer yellow fluorescence in ADPKD cells was consistent with an alteration in heterogeneity of lysosomal enzyme content in ADPKD rather than a decrease in total lysosomal number. Western analysis, metabolic labeling, and immunoprecipitation analysis confirmed decreases in the expression and synthesis of the major normal molecular immunoreactive species of β-galactosidase and cathepsins B and H in ADPKD tissue and cells but no changes in cathepsin D. In addition, ADPKD-specific high-molecular-weight species of cathepsin H were seen and abnormal forms of cathepsin B and β-galactosidase were common in ADPKD, suggesting abnormal molecular processing and posttranslational modifications. In addition, immunolocalization studies showed abnormal apical plasma-membrane localization of cathepsins B and H in ADPKD cyst epithelial cells, consistent with a protein sorting defect in ADPKD. Increased extracellular secretion of lysosomal enzymes was also measured in ADPKD cultured cells and in filter-grown epithelia shown to be predominantly directed to the basal compartment. These results demonstrate that lysosomal enzyme alterations in ADPKD may play a role in aberrant processing of the basement membrane. Alterations in the polarized secretion of lysosomal enzymes by ADPKD epithelia<em>in vitro</em>were also detected. Whereas all normal epithelia cells secreted lysosomal enzymes predominantly to the apical medium compartments, basally directed secretion was increased in all ADPKD epithelia and attained an overall reversal of polarity for cathepsins B + L. It is concluded that alterations in lysosomal enzyme function in ADPKD are the result of alterations in synthesis, molecular processing, and polarized secretion of specific enzymes and may have impact on proliferative and basement membrane abnormalities in this genetic disease. These results are consistent with a fundamental defect in protein processing, sorting, and trafficking in ADPKD.</p></div>","PeriodicalId":8837,"journal":{"name":"Biochemical and molecular medicine","volume":"60 1","pages":"Pages 8-26"},"PeriodicalIF":0.0,"publicationDate":"1997-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/bmme.1996.2542","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20023601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}