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Biomedical subjects

P Berg

Publications and source records attributed to P Berg.

At least 199 records · Page 11Linked to original sources
PubMed1983

Simian virus 40 early- and late-region promoter functions are enhanced by the 72-base-pair repeat inserted at distant locations and inverted orientations.

Tandemly repeated 72-base-pair (bp) segments located between nucleotides 107 and 250 of the simian virus 40 genome are essential for early region transcription. The functional requirement for the 72-bp repeat was supplied even when that segment was translocated to several locations distant from, and in different orientation, relative to, the promoter. Regardless of the position of the 72-bp enhancer segment, transcription was initiated at the same locations as with the normal promoter. Translocation of the 72-bp repeat segment to other sites in the genome resulted in the appearance of DNase I hypersensitivity at that site in the intranuclear viral minichromosomes. One of the translocations which did not produce enhancement of early- and late-region expression also failed to create a DNase I-hypersensitive site at the translocated 72-bp segment.

Base Sequence↗
PubMed1983

The mitochondrial adenine nucleotide translocator is an antigen in primary biliary cirrhosis.

Circulating antibodies reacting specifically with the adenine nucleotide translocator from liver mitochondria were detected in sera from 12 patients with proven primary biliary cirrhosis (PBC) by a solid phase double antibody immunoradiometric assay (IRMA). Furthermore these antibodies were absorbed with the isolated adenine nucleotide translocator from liver mitochondria. None of the sera from 20 normal individuals, four patients with anti-mitochondrial positive pseudolupus syndrome (PLE) sera (M-3) and three patients with syphilis (anti-M-l) had antibodies directed against this protein from inner mitochondrial membrane. The adenine nucleotide translocator as antigen in PBC could clearly be distinguished from the ATPase associated PBC specific M-2 antigen. With the present study, for the first time, a well characterized protein from inner mitochondrial membrane has been clearly defined as an autoantigen in primary biliary cirrhosis.

Adenosine Triphosphatases↗
PubMed1983

Transcription in vivo from SV40 early promoter deletion mutants without repression by large T antigen.

Transfection of monkey cells with recombinant plasmids containing a beta-globin coding region fused to wild-type or deleted simian virus 40 (SV40) early region promoters has allowed an analysis of the transcriptional activity of these promoters in the absence of repression by large T antigen. We find that deletion of the TATA sequence does not reduce the transcriptional effectiveness of the promoter; however, the 5' ends of the transcripts are heterogeneous rather than being restricted to the usual sites. The short GC-rich repeat sequences between nucleotides 37 and 107 and the tandemly repeated 72-bp segment between nucleotides 107 and 250 are each essential for promoter function. The GC-rich repeats are functionally redundant and probably interchangeable, since several subsets of two or three of the GC-rich segments are sufficient. One copy of the 72-bp sequence is sufficient to permit transcription. Moreover, the 72-bp repeat sequences function normally even if they are inverted relative to their normal orientation.

Antigens, Viral↗
PubMed1982

[Comparative treatment of gastroduodenal haemorrhage with secretin and cimetidine].

In a prospective multicenter study 71 patients with acute, non-arterial gastroduodenal haemorrhage from ulcers or stress lesions were treated alternately with synthetic secretin (Hoe 0690 (n = 35) or with cimetidine (n = 36). Both medications were given by infusion over 48 hours. Prior to commencement of treatment the haemorrhage was verified gastroscopically. During infusion of secretin cessation of hemorrhage was achieved without recurrence in 30 out of the 35 patients within the 48-hour infusion time and in 3 patients with recurrence within the same period. Two patients failed to obtain discontinuation of haemorrhage within the infusion period. Cimetidine led to cessation of haemorrhage without recurrence within 48 hours in 20 out of 3 patients, 3 patients had recurrences. Haemorrhages could not be arrested in 13 patients within the two-day treatment period. Differences between success of treatment among the two groups are significant (P less than 0.01) favouring secretin.

Adult↗
PubMed1982

[Consequences of vitreous loss during cataract extraction ].

An analysis of 1,959 cataract operations with intraoperative loss of vitreous body in 114 cases (5.8%) is described. The most common late complications (21% of all cases) were retinal detachments (7.9%) and secondary glaucomas (3.5%). As a consequence the authors call for regular follow-up examinations by an ophthalmologist.

Adolescent↗
PubMed1982

Regulated expression of human interferon beta 1 gene after transduction into cultured mouse and rabbit cells.

The human interferon beta 1 gene has been inserted into simian virus 40 hybrid plasmid vectors carrying the bacterial phosphotransferase gene (neo), and introduced into cultured mammalian cells by DNA transfection. A majority of the transformants resistant to the antibiotic G418 were capable of synthesizing and secreting biologically active human interferon. The neo/interferon transformants contain several copies of the transfecting DNA integrated into cellular DNA sequences. In most transformants the production of human interferon and its mRNA is induced by the addition of poly(rI) X poly(rC); by contrast, the level of neo mRNA is not increased under the same conditions. The 5' end of the human interferon mRNA produced after induction was indistinguishable from the interferon mRNA induced in human fibroblasts. This indicates that information enabling human beta 1 interferon gene to be induced by poly(rI) X poly(rC) is localized to sequences within, or 5'-proximal to, the coding sequence.

Animals↗
PubMed1982

Simian virus 40-rabbit beta-globin recombinants lacking late mRNA splice sites express cytoplasmic RNAs with altered structures.

Deletions were introduced at exon-intron boundaries in the late region of a simian virus 40-beta-globin cDNA recombinant to study the role of splicing in the formation of simian virus 40 late cytoplasmic RNAs. The recombinant was used as a wild type because it allowed characterization of mutant RNAs expressed from defective genomes in the presence of comparable RNAs contributed by the coinfecting helper virus. Removal of a 17-base pair segment at map position 0.76, which included a portion of the leader sequence implicated in the splicing of the major 16S mRNA, prevented expression of 16S-type mRNA. The same mutant accumulated cytoplasmic 19S-type mRNA, but the assortment of the 5' ends of these mRNAs differed from the assortment of the wild-type counterparts. Another mutant that lacks nucleotide sequences implicated in the splicing of the major 16S mRNA and one of the principal 19S-type RNAs accumulated a 16S-type mRNA with a previously undetected leader splice, and assortment of 19S mRNAs with new or normally underrepresented splices, and even a species of unspliced cytoplasmic 19S mRNA.

Animals↗
PubMed1982

High-efficiency cloning of full-length cDNA.

A widely recognized difficulty of presently used methods for cDNA cloning is obtaining cDNA segments that contain the entire nucleotide sequence of the corresponding mRNA. The cloning procedure described here mitigates this shortcoming. Of the 10(5) plasmid-cDNA recombinants obtained per microgram of rabbit reticulocyte mRNA, about 10% contained a complete alpha- of beta-globin mRNA sequence, and at least 30 to 50%, but very likely more, contained the entire globin coding regions. We attribute the high efficiency of cloning full- or nearly full-length cDNA to (i) the fact that the plasmid DNA vector itself serves as the primer for first- and second-strand cDNA synthesis, (ii) the lack of any nuclease treatment of the products, and (iii) the fact that one of the steps in the procedure results in preferential cloning of recombinants with full-length cDNA's over those with truncated cDNA's.

Animals↗
PubMed1982

Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.

A bacterial gene (neo) conferring resistance to neomycin-kanamycin antibiotics has been inserted into SV40 hybrid plasmid vectors and introduced into cultured mammalian cells by DNA transfusion. Whereas normal cells are killed by the antibiotic G418, those that acquire and express neo continue to grow in the presence of G418. In the course of the selection, neo DNA becomes associated with high molecular weight cellular DNA and is retained even when cells are grown in the absence of G418 for extended periods. Since neo provides a marker for dominant selections, cell transformation to G418 resistance is an efficient means for cotransformation of nonselected genes.

Animals↗
PubMed1982

Deletion mapping of DNA regions required for SV40 early region promoter function in vivo.

The SV40 early region promoter, previously localized to the DNA segment bounded by the HpaII and HindIII restriction sites (nucleotides 346 and 5171), was further defined by construction of an extensive set of deletions within this region and measurement of their effects on (a) viral DNA replication, (b) virus multiplication and the ability to complement early and late mutations, (c) transformation of rat cells, (d) large T antigen formation, and (e) the location of the 5' ends of early mRNAs. One set of mutations is represented by deletions that begin at the HpaII site and extend unidirectionally for varying lengths toward the BglI site at ori. A second set of mutants contains deletions that start at ori and extend unidirectionally for varying lengths towards the HpaII site. A third set of mutants, with deletions or duplications of various lengths and boundaries, lie between the HpaII and BglI sites. Our studies indicate the following. (a) Ori, the sequence needed for initiating SV40 DNA replication, extends from the sequences needed for initiating SV40 DNA replication, extends from the sequences needed to bind T antigen to the palindrome in site II to nucleotide 34, the late region edge of the AT block. Flanking sequences adjacent to the AT block facilitate DNA replication. (b) The SV40 early region promoter comprises two functionally distinct nucleotide sequence elements. One is flanked by nucleotides 5231 and 107, and contains the RNA initiation sites at nucleotides 5231-5237, a positioning element resembling the TATAAATA consensus sequence about 20-25 nucleotides upstream, and an RNA polymerase II recognition sequence contributed by short GC-rich sequences clustered between nucleotides 35 and 107; we refer to this as the RNA polymerase II interaction site. The second distinct sequence element is contained within each of two 72-bp segments located between nucleotides 107 and 250; the behavior of this element suggests that it may influence the accessibility of RNA polymerase II for the interaction site or the efficiency of RNA chain initiation. Large T antigen binding sites I, II, and III overlap with the putative RNA polymerase II interaction site; since large T antigen does not prevent elongation of RNA transcripts initiated upstream, T antigen probably represses early region expression by preventing RNA polymerase II binding to the promoter.

Chromosome Deletion↗
PubMed1982

The effects of diazoxide upon fetal and maternal hemodynamics and fetal brain function and metabolism.

The effects of maternal infusion of Diazoxide (D) were evaluated in two groups of experiments: Nine ewes and their five nonhypoxic fetuses; the remaining four fetuses were excluded because of the development of hypoxia prior to the study. D was directly administered intravenously (IV) to the six fetuses. In group 1 maternal and fetal observations were made at 0, 5, 15, 30, and 45 minutes. A 60-minute observation was also made for the maternal infusion studies. Maternal infusion of 5 mg/kg maternal weight significantly reduced maternal pH from the control value at 5 minutes. Maternal metabolic acidosis was indicated by concomitant significant reductions in base deficit (BD) and total bicarbonate (HCO3), both of which remained low for all subsequent time periods. Maternal lactic acid concentration also increased at 15 minutes, although not significantly. Maternal PO2 also fell significantly at 5 minutes, remaining significantly low for the remaining observations. A profound maternal hypotension was elicited at 5 minutes. All following observations for maternal blood pressure (MBP) were also significantly low. Fetal carotid arterial (FCA) pH declined significantly from the control period value at 15 minutes, remaining significantly low for the remaining observations. FCA O2% saturation (O2%) also witnessed a significant decline from baseline for all observations. FCA serum lactate level rose at 15 minutes; however, this was not deemed significant. No significant changes were seen in fetal brain metabolic rate of oxygen or glucose. Fetal electroencephalogram (EEG) revealed a definite hypoxic change in a majority of cases. In group 2, the effects of a direct fetal IV administration of 30 mg D (mean dose = 7.97 mg/kg fetal weight) were studied in six experiments. No significant changes in fetal acid base status or fetal blood pressure (FBP) were witnessed for the 45 minutes under scrutiny. Fetal heart rate (FHR) showed a short transient rise from baseline at 5 and 15 minutes, returning to normal thereafter. A slight significant decrease in O2% saturation and O2 content were seen only for the 5-minute observation. There were no significant changes of fetal brain function and metabolism observed following D administration to the fetus. The significant fetal hypoxia and acidosis witnessed through the maternal infusion but not in the direct fetal bolus administration were probably owing to profound maternal hypotension resulting in deterioration of uteroplacental perfusion and fetal "gas exchange."

Animals↗
PubMed1982

Effect of magnesium sulfate on maternal and fetal hemodynamics and fetal brain function and metabolism.

The hemodynamic effects, brain function, and metabolism of magnesium sulfate (MgSO4) were studied in seven fetal sheep experiments at 139 +/- 1.3 SE days of gestation. MgSO4 was intravenously infused to six gross-bred pregnant ewes at a rate of 4 gm/4 minutes as a loading dose, and 1.5 gm/60 minutes as a maintenance dose. There were no significant changes in maternal pH, PO2, O2 saturation, O2 content, HCO-3, glucose or lactate concentrations, or blood pressure at the end of the infusion. However, maternal heart rate decreased significantly from the mean baseline of 117 +/- 12.4 beats/minute at the end of the 60-minute infusion but recovered 30 minutes later. Uterine blood flow remained unchanged. Maternal Mg2+ levels rose significantly (P less than 0.005) at 60 minutes and remained elevated. No significant changes in fetal pH, PO2, PCO2, lactate, and Mg2+ concentration were noted during infusion. However, fetal heart rate increased significantly (P less than 0.025) at the end of infusion. Fetal hypotension was also noted at the end of the infusion (P less than 0.02). Cerebral blood flow did not change significantly throughout the experiment. However O2 consumption decreased temporarily at 15 and 30 minutes into infusion, which returned to baseline thereafter. Fetal glucose consumption and electroencephalogram remained unchanged throughout the experiment. Maternal infusion of MgSO4 does not exert any deleterious effects on the acid-base status of the mother or fetus. It increases the fetal heart rate and causes fetal hypotension, without any significant effects on the fetal brain function and metabolism.

Animals↗
PubMed1981

Antimitochondrial antibodies (AMA) in primary biliary cirrhosis. I. Separation of the PBC antigen activity from mitochondrial ATPase activity.

Antimitochondrial antibodies are found in a variety of autoimmune liver diseases, particularly primary biliary cirrhosis. The antigen against which these antibodies are directed is localized on the inner mitochondrial membrane. Earlier work suggested that this antigen was associated with the mitochondrial ATPase. However, we have succeeded in separating the enzyme activity from the antigenic activity using gel filtration and ion-exchange chromatography. Furthermore, the antigenic activity is not affected by modulators of ATPase enzymatic activity like aurovertin or oligomycin. The antigenic activity is, however, very susceptible to reagents which block thiol groups. The mitochondrial antigen, in contrast to the ATPase enzyme, is found in high amounts in brown fat mitochondria. Identification of this antigen may help to explain why specific antimitochondrial antibodies arise in the sera of patients with primary biliary cirrhosis.

Adenosine Triphosphatases↗