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Biomedical subjects

A Becker

Publications and source records attributed to A Becker.

At least 415 records · Page 23Linked to original sources
PubMed1988

Bacteriophage lambda DNA packaging. The product of the FI gene promotes the incorporation of the prohead to the DNA-terminase complex.

Lambda DNA packaging in vitro can be examined in stages. In a first step, lambda DNA interacts with terminase to form a DNA-enzyme complex, called complex I. Upon addition of proheads, in a second step, a ternary complex, complex II, containing DNA, terminase and the prohead is formed. Finally, upon addition of the rest of the morphogenetic components, complete phages are assembled. We have investigated the effect of the FI gene product (gpFI) in these reactions and found that a stimulation in phage yield is observed when gpFI is included early in the reaction, at the time when DNA, terminase and proheads interact to form complex II. Measurements of complex II formation revealed that gpFI stimulated the rate of formation of this intermediate. gpFI was further shown to stimulate the addition of proheads to preformed complexes I to give complex II, but the protein did not stimulate complex I formation.

Bacteriophage lambda↗
PubMed1988

Prediction of an ATP reactive center in the small subunit, gpNu1, of the phage lambda terminase enzyme.

The small subunit of the bacteriophage lambda terminase enzyme, the product of the phage's Nu1 gene, is shown to contain amino acid segments homologous to those present in a large number of ATPases. In keeping with these predictions, the purified protein has been found to hydrolyze ATP with a relatively low turnover number. Terminase holoenzyme is a known ATPase, and the biochemical significance of an ATP-interactive center situated in the gpNu1 subunit is discussed.

Adenosine Triphosphatases↗
PubMed1988

Optimized oligonucleotide probes for DNA fingerprinting.

The three different simple repetitive oligonucleotide probes (CT)8, (CAC)5 and (TCC)5 were hybridized to a panel of human DNAs which had been digested with the restriction endonucleases Alu I, Hinf I and Mbo I. The resulting DNA fingerprints were analyzed and different parameters calculated, such as the maximal mean allele frequency and the average number of polymorphic bands per individual. The highest number of bands was obtained after hybridization of Hinf I digested DNA with (CAC)5. The probability of finding the same band pattern as in individual A in individual B is 2 x 10(-8). The DNAs of monozygous twins show indistinguishable banding patterns and the bands are inherited according to the Mendelian laws. Thus this procedure reveals informative fingerprints that can be used for individual identification, e.g. in paternity testing and in forensic applications. In most of these experiments 32P-labelled probes were employed, yet the biotinylated oligonucleotide (GACA)4 produced results which were equivalent to those obtained by hybridization with the 32P-labelled probe (GACA)4.

Blotting, Southern↗
PubMed1988

Late stages in bacteriophage lambda head morphogenesis: in vitro studies on the action of the bacteriophage lambda D-gene and W-gene products.

The in vitro maturation of bacteriophage lambda can be divided into discrete steps. Concatemers of lambda DNA bind terminase to form complex I. This DNA-terminase complex then binds a prohead to form a ternary complex (II). Complex II in turn can be converted to infectious phage by the addition of extracts containing the products of the phage genes D, W, FII, as well as phage tails. By using in vitro complementation assays gpD and gpW have been partially purified and their interactions with complex II studied. gpD can bind to complex II in vitro to form a new complex (III) which can be isolated by sedimentation on neutral sucrose gradients. This complex requires only the addition of gpW, gpFII, and phage tails to form mature phage particles. The sedimentation of complex III is virtually identical to that of complex II; however, the resistance of the former to inactivation by DNase is higher, likely due to the partial packaging of the DNA. In similar experiments it was shown that gpW cannot bind to complex II but can effectively interact with complex III. This latter reaction converts complex III to a DNase-resistant form which sediments in a manner identical to that of full phage heads (complex IV). After isolation of the complex IV only gpFII and tails are required for mature phage formation in vitro. gpW is a heat-stable protein of molecular weight approximately 10,000.

Bacteriophage lambda↗
PubMed1988

Formulation of vaccine adjuvant muramyldipeptides (MDP). 1. Characterization of amorphous and crystalline forms of a muramyldipeptide analogue.

A relatively nonhygroscopic crystalline form of the glycopeptide, N-acetylmuramyl-L-alpha-aminobutyryl-D-isoglutamine (I), containing approximately one molecule of water was prepared from amorphous material. The crystalline material, consisting of a mixture of the alpha and beta anomers, exhibited better physical and chemical stability than the lyophilized amorphous material. The alpha/beta-anomer ratios of I in both the crystalline and the amorphous state were approximately equal but different from that in solution.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1988

Infrequent genomic rearrangement and normal expression of the putative RB1 gene in retinoblastoma tumors.

Retinoblastoma (RB) tumors develop when both alleles of a gene (RB1) are mutated and unable to function normally. Recently, Friend et al. [S. H. Friend, R. Bernards, S. Rogelj, R. A. Weinberg, J. M. Rapaport, D. M. Albert, and T. P. Dryja, Nature (London) 32:643-646, 1986] reported the cloning of a gene, 4.7R, with some properties expected for the RB1 gene, namely, a high frequency (30%) of genomic rearrangements in tumors and absence of message in all RB tumors examined. To extend the characterization of this gene, we used 4.7R probes to search for genomic rearrangements of DNA and to study the expression of the 4.7R gene in RB tumors, osteosarcoma (OS) tumors arising in RB patients, and other normal and malignant tissues. In 34 previously unreported RB and OS tumors arising in RB patients, we observed only four (12%) with genomic abnormalities. Transcripts of 4.7R were present in 12 of 17 RB tumors, 2 of 2 OS tumors, and all non-RB tumors and normal tissues tested. We were unable to confirm the high frequency of truncated messages of 4.7R in RB tumors reported by Lee et al. (W. H. Lee, R. Bookstein, F. Hong, L. J. Young, J. Y. Shaw, and E. Y. Lee, Science 235:1394-1399, 1987) and Fung et al. (Y. K. Fung, A. L. Murphree, A. Tang, J. Qian, S. H. Hinrichs, and W. F. Benedict, Science 236:1657-1661, 1987) but did confirm the presence of a truncated transcript in the RB cell line Y79. Of the RB and RB-related OS tumors which appeared normal on Southern blots, 2 of 26 or 12% had abnormal transcripts, giving a combined frequency of 22% abnormalities in the 4.7R gene detectable by Southern and Northern (RNA) blot analyses.

Gene Expression Regulation↗
PubMed1988

[Etomidate versus etomidate and hydrocortisone for anesthesia induction in abdominal surgical interventions].

The imidazole derivative etomidate has been shown to block (reversibly) adrenocortical steroid synthesis. Long-term sedation with etomidate has been associated with adrenocortical insufficiency and increased mortality in severely ill patients. The significance of adrenocortical blockade after a single induction dose of etomidate remains a matter of debate. This study was designed to analyze the role of glucocorticoid deficiency after a single induction dose of etomidate for major surgery. In a prospective controlled double-blind study, 20 consecutive patients scheduled for colorectal surgery were randomly allocated to either hydrocortisone substitution or placebo (glucose 5%). Hydrocortisone was given as a continuous infusion in a dose of 100 mg dissolved in 5% glucose over 10 h. We combined general anesthesia and epidural anesthesia (L3-4) using bupivacaine (0.5%). Induction of anesthesia: etomidate (0.2-0.3 mg/kg), fentanyl (0.1-0.2 mg), pancuronium (2 mg), and succinyl-choline (1.0-1.5 mg/kg), with endotracheal intubation and mechanical ventilation. Anesthesia was maintained at N2O/O2 2:1 and fluothane (0.4-0.6 vol%). At the end of surgery patients were extubated after oxygenation. In all patients blood pressure, heart rate, central venous pressure, and ECG were monitored continuously, both intra- and postoperatively. During induction, patients received 1,000 ml 0.9% NaCl, followed by continuous administration of 0.9% NaCl, 6 ml/kg per hour intraoperatively and 40 ml/kg per 24 hours post-operatively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Insufficiency↗
PubMed1988

Total insulin levels in type 1 diabetic patients with insulin antibodies and their effect on insulin requirement and metabolic control.

In 158 type 1 diabetic patients the relations between insulin antibodies and total insulin and between total insulin and insulin requirement, mean blood glucose, post-prandial change in blood glucose, mean deviation factor of blood glucose, M-value and frequency of hypoglycemias were investigated. Forty-eight patients were antibody negative (antibody level less than or equal to 0.4%) and 110 patients had various levels of insulin antibodies and elevated levels of total insulin, but not of free insulin. After having made all necessary efforts to stabilize metabolism, parameters of metabolic control were determined. No effect of the elevated total insulin levels in patients with insulin antibodies could be demonstrated on insulin requirement, mean blood glucose, postprandial rise in blood glucose, mean deviation factor of blood glucose and M-value. The efforts needed to reach acceptable blood glucose levels were not greater in patients with elevated total insulin. The frequency of hypoglycemias during the night was increased in patients with total insulin levels higher than 2 mmol/l which, although not statistically significant, may be of clinical relevance.

Adolescent↗
PubMed1987

Follistatin specifically inhibits pituitary follicle stimulating hormone release in vitro.

Two forms of purified follistatin, a single-chain polypeptide of mol wt 35,000 (35 Kd) protein, and a related molecule of mol wt 32,000 (32 Kd), which differs from the 35 Kd form in glycosylation or carboxyl terminal truncation, specifically inhibit the release of immunoreactive FSH by primary cultures of rat pituitary cells. Both forms of follistatin and inhibin-A give similar dose-response curves, with identical slopes and maximal effects, suggesting that they may all act through the same mechanism on the pituitary cells. The median effective dose (ED50) of each of the follistatins is 6.2-7.3 ng/ml (1.8 x 10(-10) M), which corresponds to approximately 1/3 of the potency of inhibin. The effect of 35 Kd or 32 Kd follistatin is highly specific for suppressing the release of immunoreactive FSH since there is no demonstrable concomitant effect on the secretion of other pituitary hormones. The effect of follistatins, like that of inhibins, is different from that of the hypothalamic hypophysiotropic factors, requiring greater than or equal to 18 h of incubation in a pituitary monolayer culture system to demonstrate. Coincubation of inhibin and follistatin shows an additive effect in the suppression of FSH release. Pituitary cells exposed to follistatin have significantly less depletion of intracellular FSH (0.01) than those treated with inhibin, indicating that follistatin may act primarily on the suppression of FSH release rather than on both release and synthesis of FSH, as is the case with inhibin.

Animals↗
PubMed1987

High-performance liquid affinity chromatography of liver plasma membrane proteins.

Plasma membrane proteins from liver were analysed by concanavalin A affinity and immunoaffinity high-performance liquid chromatography. In the method, four peptide bands with apparent molecular weights of 140,000, 120,000, 80,000 and 60,000 could be isolated. In the second method, with two immobilized monoclonal antibodies, two corresponding antigens--the membrane proteins dipeptidyl-peptidase IV and GP 110--could be highly purified from plasma membrane extract with good yield in only one step.

Animals↗
PubMed1987

Vitamin K in the newborn: influence of nutritional factors on acarboxy-prothrombin detectability and factor II and VII clotting activity.

The incidence of acarboxy-prothrombin and the clotting activity of factors II and VII were evaluated on the fifth day of life in 183 healthy newborns, who had received no vitamin K prophylaxis. Acarboxy-prothrombin was detected in 93/183 newborns. All acarboxy-prothrombin-negative babies had factors II and VII clotting activities above 25% whereas a great variability was observed in acarboxy-prothrombin-positive babies: 21/93 had factor II and 14/93 had factor VII activities below 25%. Seventy-two of the acarboxy-prothrombin-positive babies had normal factor II and VII clotting times on the fifth day of life. These babies must be suspected to have had vitamin K deficiency on one of the first 4 days, as acarboxy-prothrombin has a 50% disappearance rate of 50 h. Acarboxy-prothrombin was mainly observed in breast-fed infants (84/122) and only rarely detectable in infants receiving supplementary (7/44) or exclusive formula feeding (2/17). The type of milk feeding however might be less important for the babies' vitamin K supply than the actual milk intake. All acarboxy-prothrombin-positive babies had received small amounts of milk on the first 4 days of life. In those with low factor II and VII clotting activities the milk intake was low throughout the first 4 days of life, whereas babies with acarboxy-prothrombin and and normal clotting activities had increased their milk intake to more than 100 ml on the third and fourth day of life. Recommendations for vitamin K prophylaxis in newborns should be given with regard to the feeding on the first days of life.

Biomarkers↗
PubMed1987

Secretion of follicle-stimulating hormone and production of inhibin are reciprocally related.

The production of inhibin in cultured granulosa cells from immature hypophysectomized, estrogen-treated rats and Sertoli cells from normal animals was determined by a specific radioimmunoassay using an antiserum against a synthetic replicate of [Tyr30]inhibin alpha-chain-(1-30). The amount of immunoreactive inhibin detected in the spent media of these cells is in proportion to the density of cells plated and the concentration of exogenously added follicle-stimulating hormone (FSH). In the presence of the estrogen precursor androstenedione (10(-7) M), FSH, but not luteinizing hormone, produced a dose-dependent increase in inhibin during 2-day culture of granulosa cells. In the absence of the estrogen precursor, similar but somewhat diminished inhibin production in responding to FSH was observed. Exogenously added estrogen potentiated the FSH-mediated release of inhibin in the absence of androstenedione. Neither androstenedione nor estradiol added to the cultured Sertoli cells had effect on inhibin production. A preparation of pure inhibin isolated on the basis of an in vitro bioassay and characterized chemically specifically suppressed serum FSH but not luteinizing hormone, when it was injected (24 micrograms per injection, two injections) into acutely ovariectomized rats. Thus, inhibin secreted by the granulosa and Sertoli cells specifically suppresses the secretion of pituitary FSH, and in turn FSH is primarily responsible for the inhibin production in these gonadal cells, as in a classical negative-feedback relationship.

Androstenedione↗
PubMed1987

Acarboxyprothrombin concentration [corrected] after oral prophylactic vitamin K.

The effect of prophylaxis with oral vitamin K (1 mg vitamin K1 given with the first feed) on the rate of detection of acarboxyprothrombin (PIVKA II) and factor II clotting concentration [corrected] were analysed. Introducing such prophylaxis reduced the rates of detection of PIVKA II concentration [corrected] on day 5 from 48% to zero. None of the babies given prophylaxis had factor II clotting concentration [corrected] below 40%, compared with 34 of 95 babies not given prophylaxis. This study has important implications in the prophylaxis of both classical and late onset haemorrhagic disease of the newborn.

Administration, Oral↗