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Biomedical subjects

G Link

Publications and source records attributed to G Link.

At least 109 records · Page 6Linked to original sources

[Morphologic comparison of stroma rich and stroma poor tumors in the agar colony-forming test].

In the study reported here the morphologic relationship between the original tumor and a colony cultured in vitro was investigated in stroma-rich and stroma-poor tumors. The colonies, cultured in a double-layer agar system were marked with graphite and bedded in paraffin after centrifuging. Subsequent preparation of the microtome sections was in accordance with customary histologic methods. Taking an ovarian carcinoma as an example, far-reaching agreement between the original histologic findings and the section through the colony in the light-microscopic image was demonstrated. In contrast, the tumor cell colonies in mammary carcinomas, which are typically representative of stroma-rich tumors, generally showed less similarity to the histologic structure of the mother tumor, since the formation of connective tissue cannot be induced due to the lack of mesenchymal elements in the agar culture. However, under comparable in vivo and in vitro conditions, e.g., tumor manifestation in performed cavities, there was, also with mammary carcinomas, a similar morphologic analogy between the original tumor and the clone, as in cell-rich neoplasms. The possibility of identifying malignancy directly in colonies and knowledge of their morphologic relationship to the primary tumor increase the relevance of chemosensitivity testing in colonization tests.

Breast↗

Characterization of transcriptionally active DNA-protein complexes from chloroplasts and etioplasts of mustard (Sinapis alba L.).

DNA-protein complexes that are capable of RNA synthesis in vitro (transcriptionally active chromosomes) were isolated from both chloroplasts and etioplasts of mustard (Sinapis alba L.) seedlings. Analyses of the polypeptide pattern of these complexes indicate that they comprise a specific subset of plastid proteins, distinct from the overall pattern of either the soluble or membrane-bound plastic proteins. DNA-protein complexes from the two plastid types have polypeptides in common. However, at least several polypeptides are highly enriched in either the chloroplast or the etioplast DNA-protein complex. The EcoRI restriction endonuclease fragments of the DNA associated with the complexes from either plastid type are the same. They are identical with the fragments obtained from highly purified chloroplast DNA. The transcriptional activity of the chloroplast complex is more than ten times higher than the activity of the etioplast complex. However, the complexes from either plastid type are capable of transcribing DNA regions containing genes for both the plastid rRNAs and for plastid proteins. In vitro transcripts were found to hybridize not only to DNA regions for mature in vivo RNA but also to adjacent regions, indicating synthesis of precursor RNA sequences by the transcriptionally active chromosomes.

Chemical Phenomena↗

Combined use of zinc protoporphyrin (ZPP), mean corpuscular volume and haemoglobin measurements for classifying microcytic RBC disorders in children and young adults.

The diagnostic potential of the combined use of zinc-protoporphyrin (ZPP), mean corpuscular volume (MCV) and haemoglobin measurements for discriminating between iron deficiency anaemia, beta-thalassaemia minor and lead poisoning has been studied. Lead poisoning could be identified by ZPP greater than 50 micrograms/dl in the presence of normal MCV or ZPP greater than 150 micrograms/dl in the presence of microcytosis (MCV less than 80 fl) with a sensitivity of 97% and specificity 94%. Beta-thalassaemia minor was identified by the coexistence of microcytosis and ZPP less than 50 micrograms/dl with a sensitivity of 91% and specificity 79%. Iron deficiency anaemia defined by the combination of microcytosis and ZPP ranging from 50 to 150 micrograms/dl was identified with a sensitivity of 95%, but the specificity was only 51%, with many of the patients overlapping with thalassaemia minor. This problem did not exist in iron-deficiency anaemia with haemoglobin less than 10 g/dl as at that range no patients with uncomplicated thalassaemia minor have been encountered. A great advantage of the combined use of ZPP, MCV and haemoglobin for the initial screening of microcytic anaemia is its ease of performance and low cost. However, this information should only be regarded as presumptive evidence of disease, requiring subsequent confirmation by appropriate direct measurements such as transferrin saturation, serum ferritin, haemoglobin electrophoresis, or blood lead determinations.

Adolescent↗

Heart cells in culture: a model of myocardial iron overload and chelation.

The effect of iron loading and chelation was studied in heart cell cultures obtained from newborn rats. Radioactive iron uptake per 2 X 10(6) cells/24 hr was 3.8% for 59Fe-transferrin, 15.8% for 59Fe-ferric ammonium citrate (FeAC) at 20 micrograms Fe/ml in 20% serum, and 37.1% for 59FeAC at 20 micrograms Fe/ml in serum-free medium. About one third of the cellular radioactive iron was in ferritin and the rest in an insoluble lysosomal fraction. Iron uptake was almost completely inhibited by reducing the incubation temperature from 37 degrees C to 10 degrees C. Intracellular concentrations of malonyldialdehyde (MDA) were doubled after 15 minutes of iron loading and reached maximal concentrations at 3 hours. Conversely, iron mobilization by deferoxamine at concentrations ranging from 0.025 mmol/L to 0.3 mmol/L resulted in normalization of cellular MDA concentrations, in direct proportion to the amounts of iron removed. These findings indicate that cultured myocardial cells are able to assimilate large amounts of nontransferrin iron and that iron uptake and mobilization are associated with striking changes in lipid peroxidation as manifested by the respective increase and decrease in cellular MDA concentrations.

Animals↗

Structure of the chloroplast gene for the precursor of the Mr 32,000 photosystem II protein from mustard (Sinapis alba L.).

The nucleotide sequence of the mustard chloroplast gene for the precursor of the Mr 32,000 photosystem II protein is presented. A comparison with the corresponding genes from spinach and Nicotiana debneyi (14) reveals less than 5% nucleotide divergence in the coding region. The derived protein of mustard differs from the corresponding proteins by three amino acid positions at the C-terminus. We have defined the presumed transcription start and termination sites of the mustard gene. Upstream from the start site are sequences typical of a prokaryotic promoter and, also, a sequence that resembles the eukaryotic 'TATA' box. A search for intrastrand base pairing revealed stem-loop secondary structure at the transcription start and termination sites and in the region preceding the presumed promoter. This latter region is a 69-base-pair sequence element unique to the 5'flanking sequence of the mustard gene.

Base Sequence↗

DNA sequence requirements for the accurate transcription of a protein-coding plastid gene in a plastid in vitro system from mustard (Sinapis alba L.).

A nuclease-treated plastid extract from mustard (Sinapis alba L.) allows efficient transcription of cloned plastid DNA templates. In this in vitro system, the major runoff transcript of the truncated gene for the 32 000 mol. wt. photosystem II protein was accurately initiated from a site close to or identical with the in vivo start site. By using plasmids with deletions in the 5'-flanking region of this gene as templates, a DNA region required for efficient and selective initiation was detected 28-35 nucleotides upstream of the transcription start site. This region contains the sequence element TTGACA, which matches the consensus sequence for prokaryotic ;-35' promoter elements. In the absence of this region, a region 13-27 nucleotides upstream of the start site still enables a basic level of specific transcription. This second region contains the sequence element TATATAA, which matches the consensus sequence for the ;TATA' box of genes transcribed by RNA polymerase II (or B). The region between the ;TATA'-like element and the transcription start site is not sufficient but may be required for specific transcription of the plastid gene. This latter region contains the sequence element TATACT, which resembles the prokaryotic ;-10' (Pribnow) box. Based on the structural and transcriptional features of the 5' upstream region, a ;promoter switch' mechanism is proposed, which may account for the developmentally regulated expression of this plastid gene.

Journal Article↗

Development and evaluation of the improved iron chelating agents EHPG, HBED and their dimethyl esters.

The phenolic EDTA analogues ethylenediamine-N,N'-bis-(2- hydroxyphenylglycine ) ( EHPG ), N,N'-bis(2-hydroxybenzyl)-ethylenediamine diacetic acid ( HBED ), and their respective dimethyl esters ( dimethylEHPG and dimethylHBED ) were studied in hypertransfused rats. Radioiron bound to these compounds was cleared mainly by the liver and excreted in the bile. After a single 40 mg i.m. injection, the percentage of radioiron removed from 59Fe-ferritin-labelled hepatocytes and excreted in the bile was 4% in untreated controls, 24% for desferral , 42% for dimethylEHPG , 58% for EHPG , 63% for HBED , and 80% for dimethylHBED . DimethylHBED combines oral effectiveness with superior chelating ability, selective hepatocellular action, and low apparent toxicity. It may represent a significant advance in the development of new iron chelating drugs.

Administration, Oral↗

Iron depletion and blood lead levels in a population with endemic lead poisoning.

The effect of iron depletion on blood lead levels was studied in a group of 558 schoolchildren aged 10 to 18 residing in two West Bank Arab villages with a high prevalence of excessive lead exposure and clinical lead poisoning. Thirty percent of the subjects studied had whole-blood lead levels greater than 30 micrograms/dl, and 45% had whole-blood zinc erythrocyte protoporphyrin greater than 50 micrograms/dl. The percentage of abnormal blood lead values in subjects with normal serum ferritin and transferrin saturation (31%) was similar to that in subjects with one (28%) or two (25%) abnormal iron parameters. Likewise, there was no correlation between serum ferritin and blood lead levels in individual patients (r = 0.059). Normal distribution plots and mean levels of blood lead (27.3 micrograms/dl) were identical in the iron-deficient and iron-replete groups. These findings indicate that iron depletion does not affect blood lead levels and suggest that in man, unlike in experimental animals, iron depletion may not have a significant effect on lead absorption.

Adolescent↗

Phenolic ethylenediamine derivatives: a study of orally effective iron chelators.

Of 35 potential iron chelators screened for in vivo activity in rats, a group of phenolic compounds with excellent chelating properties were identified. These included N,N'-ethylene-bis(o-hydroxyphenylglycine) (EHPG), N,N'-Bis(o-hydroxybenzyl)-ethylenediamine diacetic acid (HBED), and their respective dimethyl esters (dmEHPG and dmHBED). All four phenolic compounds produced a marked increase in the fecal excretion of hepatocellular radioiron. This amounted to 42% of total body radioactivity with dmEHPG, 58% with EHPG, 63% with HBED, and 80% with dmHBED after a single injection of 40 mg/animal. At a dose of 5 mg/animal, EHPG, HBED, and dmHBED were 9, 12, and 15 times more potent, respectively, than deferoxamine. Both dimethyl esters showed significant oral activity; oral dmEHPG retained 1/3 and dmHBED retained 2/3 of the effect of the same dose given by intramuscular injection. The ester dmHBED combines oral effectiveness with superior chelating ability, selective hepatocellular action, and low apparent toxicity. It may represent a significant advance in the development of new iron chelating drugs.

Administration, Oral↗

Syntheses of iron bis(pyridoxal isonicotinoylhydrazone)s and the in vivo iron-removal properties of some pyridoxal derivatives.

Pyridoxal isonicotinoylhydrazone (PINH; 1) and its isomeric O-acetates (E and Z) were synthesized and complexed with ferrous ions to afford the hitherto unisolated chelates iron(II) bis(pyridoxal isonicotinoylhydrazone)s (11) and iron(II) bis(O-acetylpyridoxal isonicotinoylhydrazone)s (12). The analytical and spectroscopic data of the new coordination compounds are presented. In addition, a series of imino derivatives of pyridoxal of structures 2-3 and 5-10 have been prepared and tested in vivo as chelators of storage iron, and the cumulative net excretion of radioiron in urine and in feces was estimated. This study reestablishes that PINH is a potent iron chelator in vivo comparable in efficiency with parenteral desferrioxamine (DF) and indicates that it requires further attention.

Animals↗

A rapid and sensitive ELISA for serum ferritin employing a fluorogenic substrate.

A fluorescent enzyme-linked immunosorbent assay is described for the rapid measurement of serum ferritin. Increased sensitivity was achieved by using 4-methyl-umbelliferyl-beta-D-galactopyranoside as the substrate for beta-galactosidase coupled to the purified antiferritin antibody. Further enhancement of the specific antigen-antibody reaction was attained by the addition of 4% polyethylene glycol 6000 to the antiferritin-beta-galactosidase conjugate. The procedure is performed in microELISA plates. These modifications of the method permit the measurement of serum ferritin at concentrations ranging from 0.25 to 50 microgram/liter with a coefficient of variation of 8% or less. The entire procedure is performed at ambient temperature and is completed within one working day. The cost of the assay is less than 10% of the immunoradiometric assay for serum ferritin.

Animals↗

Evaluation of iron-chelating agents in an in vivo system: potential usefulness of EHPG, a powerful iron-chelating drug.

Fifteen compounds with a high affinity to ferric iron have been screened for in vivo iron-chelating efficiency in a rat model. One of the most potent of these drugs was ethylenediamine-N,N'-bis(o-hydroxyphenylglycine) (EHPG). EHPG-induced iron excretion was up to 8 times higher than iron excretion induced by identical doses of desferrioxamine (DF). Studies employing selective radio-iron probes of reticuloendothelial and parenchymal iron stores showed that although EHPG is able to interact with both storage iron compartments, its effect on parenchymal iron is much more pronounced. Unlike DF which has two alternative routes of excretion, EHPG-induced iron excretion is restricted mainly to the gut. Although EHPG seems to be superior to DF in both its chelating efficiency and preferential interaction with hepatic parenchymal iron stores, information on its in vivo toxicity is at present insufficient and it cannot yet be recommended for clinical use.

Animals↗

Cloning and mapping of the chloroplast DNA sequences for two messenger RNAs from mustard (Sinapis alba L.).

Restriction fragments of chloroplast (cp)DNA from mustard leaves were cloned in E. coli using pBR 322 as the vehicle. Cloned fragments containing the structural sequences for two polypeptides of 56,000 and 35,000 daltons were selected by a mRNA hybridization-translation procedure. The cloned mustard genes are structurally related to chloroplast genes from maize. They hybridize to the maize mRNAs for the large subunit of ribulosebisphosphate carboxylase and for the 34,500 dalton precursor to a membrane protein. The coding sequence on mustard cpDNA for the 56,000 dalton polypeptide is colinear with a 1,500 base pair transcript, and the sequence for the 35,000 dalton polypeptide is colinear with a 1,220 base pair transcript.

Base Sequence↗

Differential transcription in vivo and in vitro of two adjacent maize chloroplast genes: The large subunit of ribulosebisphosphate carboxylase and the 2.2-kilobase gene.

The transcription of cloned maize plastid DNA sequences in vitro by maize plastid DNA-dependent RNA polymerase has been studied to expose the roles of the enzyme, polypeptide cofactors, and DNA sequences in the regulation of gene expression. The 4.35-kilobase pair BamHI fragment 9 carries the maize plastid gene for the large subunit of ribulosebisphosphate carboxylase and part of the gene for a 2.2-kilobase RNA. These two genes are separated by approximately 330 base pairs and are transcribed divergently. Transcripts of the gene for the large subunit of ribulosebisphosphate carboxylase are abundant in bundle sheath cells of maize leaves and we show here that transcripts of the 2.2-kilobase RNA gene are present in both mesophyll cells and the adjacent bundle sheath cells. In vitro, in the presence of the S factor, maize chloroplast DNA-dependent RNA polymerase produces a transcript of the gene for the large subunit of ribulose-bisphosphate carboxylase with a 5' terminus like that of the corresponding mRNA isolated from plastids, transcribes chloroplast DNA sequences of Bam fragment 9 in a chimeric plasmid in preference to the vehicle RSF 1030 and, in a ratio of 3:1, preferentially transcribes the gene for the large subunit of ribulosebisphosphate carboxylase over the 2.2-kilobase RNA gene from supercoiled chimeric plasmid DNA.

Journal Article↗

Mechanism of in vivo iron chelation by pyridoxal isonicotinoyl hydrazone and other imino derivatives of pyridoxal.

The source of iron chelated in vivo by the new iron-chelating agent PIH and its mechanism of excretion have been studied in normal and hypertransfused rats. PIH is able to chelate iron from both parenchymal and RE iron stores. Unlike DF, which promotes both urinary and fecal iron excretion, in this model PIH-induced iron excretion is limited almost entirely to the gut. Response to PIH is directly related to dosage, and oral doses ranging from 125 to 500 mg/kg/day are well tolerated. Six additional imino derivatives of pyridoxal have been studied, but none of these new compounds was as effective as PIH. Our study indicates that oral PIH is comparable in efficiency with parenteral DF and is of potential usefulness in the management of iron overload.

Animals↗

Sizes, locations, and directions of transcription of two genes on a cloned maize chloroplast DNA sequence.

mRNA for the large subunit (LS) of ribulose-1,5-bisphosphate carboxylase [3-phospho-D-glycerate carboxylase (dimerizing), EC 4.1.1.39] of Zea mays is complementary to an uninterrupted 1600-base-pair-long chloroplast DNA sequence that has been mapped precisely within the 4350-base-pair-long chloroplast DNA fragment Bam 9 to which it had been traced earlier [Bedbrook, J. R., Coen, D. M., Beaton, A. R., Bogorad, L. & Rich, A. (1979) J. Biol. Chem. 254, 905-910]. An additional 1400-base-pair-long uninterrupted region that is colinear with a chloroplast RNA has been detected on Bam 9. The transcript from this region is part of a 2200-nucleotide-long RNA. The remainder of the DNA sequence for the 2200-base-pair RNA maps outside Bam 9. The 1600-base-pair LS gene and the gene for the 2200-nucleotide transcript are close to one another. They are separated by an untranscribed intercistronic "gap" about 330 base pairs long. These two closely packed genes are inverted on the chromosome-i.e., their 3' termini are at opposite ends of the untranscribed gap and they map on opposite strands.

Journal Article↗

Comparison of Large Subunits of Type II DNA-dependent RNA Polymerases from Higher Plants.

Two-dimensional tryptic mapping of (125)I-labeled polypeptides has been employed to compare the large subunits of type II DNA-dependent RNA polymerases from maize, parsley (Petroselinum sativum), and wheat. Maps of the 220 kilodalton (kd) and 140 kd subunits from wheat RNA polymerase II differ from those of the corresponding subunits from parsley enzyme II. The 180 kd subunits from maize and parsley type II enzymes also yield dissimilar tryptic maps. Thus, despite similarities in molecular mass, the large subunits of wheat, parsley, and maize type II RNA polymerases are unique to each individual plant species.

Journal Article↗