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Biomedical subjects

A Becker

Publications and source records attributed to A Becker.

At least 325 records · Page 18Linked to original sources
PubMed1995

In situ mitral valve stabilization with glutaraldehyde.

Mitral valve repair in the young rheumatic patient carries a high reoperation rate due to progression of the disease. In an attempt to halt or at least slow down this process, the possibility of fixing in situ the valve tissues with glutaraldehyde was explored. Six weanling sheep underwent tanning of their anterior mitral leaflet for two minutes with 0.5% buffered glutaraldehyde. The non-treated posterior mitral leaflet served as control. The animals were sacrificed at varying intervals between 2.5 and 6 months. At sacrifice, Doppler echocardiography and hemodynamic studies were done. The leaflets were subjected to histopathologic examination and calcium and glutaraldehyde contents were estimated. Glutaraldehyde treatment of the anterior leaflet caused thickening of the cusp and chordae associated with partial devitalization of its core tissue, partial loss of endothelium and intense fibrocellular reaction with abundant elastic fibers without altering its functional integrity. It did not induce calcification. There were no detectable levels of glutaraldehyde at explantation. The posterior mitral leaflets were normal. Although the absence of calcification and partial viability of the tissue are encouraging, it does not necessarily follow that this treatment would arrest progression of the underlying disease. This process may have clinical application in the future, but it is not yet recommended.

Animals↗
PubMed1995

Major cat allergen (Fel d I) levels in the homes of patients with asthma and their relationship to sensitization to cat dander.

BACKGROUND: Exposure and sensitization to the major cat allergen, Felis domesticus allergen I (Fel d I), significant causes of allergic respiratory disease. Many patients who are allergic to cats, however, do not own a cat and there is not an obvious source of allergen in their home environment. OBJECTIVE: We investigated the levels of Fel d I in dust from homes of 120 subjects with asthma in two climatologically diverse Canadian cities (Vancouver and Winnipeg). Fel d I levels were related to exposure to cats as well as to skin reactivity to cat dander. METHODS: Dust samples from bedroom floors and mattresses were collected in four different seasons and Fel d I content was determined by 2-site monoclonal antibody based-ELISA. RESULTS: Although only 18 patients (15%) were cat owners, detectable levels of Fel d I were found on at least one occasion in all homes. The geometric mean concentration of Fel d I on floors was 1.15 micrograms/g of dust (range 0.07 to 26.3 micrograms/g) and on mattresses 0.89 micrograms/g (range 0.01 to 17.4 micrograms/g). Seasonal variation of Fel d I levels was only observed in Winnipeg, where Fel d I concentrations were highest in the winter and spring compared with either summer (P < .05) or autumn (P < .005). The highest Fel d I levels were found in homes with a cat (P < .05), however, rather high levels were also found in homes of patients who did not have a cat but visited others with cats. Cat dander was the most frequent sensitizer (60%) in these patients but no correlation was found between the size of the wheal induced by cat dander extract and Fel d I levels in dust samples. CONCLUSION: Cat allergen was universally found in homes of asthmatic patients and this may explain the high frequency of cat sensitization among patients with asthma in these two cities. A seasonal variation in cat allergen was observed in Winnipeg with no variation in Vancouver.

Adolescent↗
PubMed1994

Chromosome end formation in phage lambda, catalyzed by terminase, is controlled by two DNA elements of cos, cosN and R3, and by ATP.

The terminase enzyme of phage lambda is a site-specific endonuclease that nicks DNA concatemers to regenerate the 12 nucleotide cohesive ends of the mature chromosome. The enzyme's DNA target, cos, consists of a nicking domain, cosN, and a binding domain, cosB. cosB, situated to the right of cosN, comprises three 16 bp repeat sequences, R1, R2 and R3. A similar sequence, R4, is present to the left of cosN. It is shown here that terminase has an intrinsic specificity for cosN which is independent of the R sites. The interaction with cosN is mediated by binding to target sites that include 12 bp on the 5', and 2-7 bp on the 3' side of the nick. Of the four R sites, only R3 is required for the proper formation of ends. When R3 is present, an ATP-charged terminase system correctly catalyzes the production of staggered nicks in cosN, at sites N1 and N2 on the bottom and top strands, respectively. When ATP is omitted, the bottom strand is nicked incorrectly, at the site Nx, 8 bp to the left of N1. If R3 is removed or disabled by a point mutation, nicking in cosN becomes dependent upon ATP but, even in the presence of ATP, bottom strand nicking is divided between sites N1, the correct site, and Nx, the incorrect one. Thus, R3 is an important regulatory element and must reside in cis in respect to cosN. Furthermore, cosN substrates bearing point mutations at N1 and N2 are nicked at sites Nx and Ny, 8 bp to the left of N1 and N2, respectively. When R3 is present and ATP is added, nicking is redirected to the N1 and N2 positions despite the mutations present. Thus, terminase binding to R3, on one side of cosN, regulates the rotationally symmetric nicking reactions on the bottom and top strands within cosN.

Adenosine Triphosphate↗
PubMed1994

The lambda terminase enzyme measures the point of its endonucleolytic attack 47 +/- 2 bp away from its site of specific DNA binding, the R site.

lambda terminase is an ATP-interactive, site-specific endonuclease comprising the products of lambda genes Nu1 and A. Terminase binds to cos, at the junction of two chromosomes in a concatemer, catalyzes cos cleavage and initiates the packaging of lambda DNA into proheads. cos consists of a nicking domain, cosN, where terminase cleaves to regenerate the 12 nucleotide cohesive ends of mature lambda chromosomes and a binding domain, cosB, where terminase binds to 16 bp repeat sequences called R3, R2 and R1. Evidence is presented that terminase is a single-strand endonuclease that can nick DNA by one of two mechanisms, both of which require ATP. (i) When bound to any R site, terminase nicks the strand which, within that R site, is purine-rich; the position of this nick is 47 +/- 2 nucleotides away from the mid-point of that R site, measured in the 3' direction; (ii) enzymes that are not bound to R sites nick DNA within certain specific sequences that resemble cosN half sites. These two modes of action are nicely combined for the R3-bound protomer that nicks the bottom strand at position N1 in cosN since the interval between N1 and the R3 midpoint is 47 nucleotides. Within cosN, the bottom and top strand nicks are generated by a rigid protein couple with a 2-fold rotational symmetry. The location of both of these nicks, however, is gauged asymmetrically from R3, 47 nucleotides away. Again, R1 and R2 are separated by 47 bp and orient bound protomers towards each other but, unless the DNA between these R sites is lengthened, the enzymes do not nick, indicating an inhibitory gpA-gpNu1 apposition.

Bacteriophage lambda↗
PubMed1994

Naloxone ameliorates the learning deficit induced by pentylenetetrazol kindling in rats.

Endogenous opioid peptides modulate and regulate processes of central excitability. Furthermore, opioids are thought to interfere with processes of learning and memory storage. In order to study the effects of endogenous opioids on both processes we injected in the course of development of pentylenetetrazol kindling the opiate receptor antagonist naloxone, and tested the animals afterwards in a shuttle-box task. It was found that naloxone pretreatment had dissociative effects. There was no effect on seizure outcome, whereas the learning deficit was ameliorated in the kindled group. The data suggest that endogenous opioid peptides contribute to the learning deficit found in pentylenetetrazol-kindled rats.

Animals↗
PubMed1994

Identification of a putative membrane-interacting domain of CTP:phosphocholine cytidylyltransferase from rat liver.

A putative membrane-interacting domain of CTP:phosphocholine cytidylyltransferase (CT) was identified using two peptide-specific antibodies. One antibody (SA2) was raised against the N-terminus of CT (amino acid residues 1-17) and the other antibody (SA209) against an alpha-helical domain of the enzyme (amino acid residues 247-257). Both antibodies quantitatively immunoprecipitated CT from rat liver cytosol and showed specificity towards CT when octylglucoside extracts of rat liver cytosol were assessed by Western blot analysis. However, further experiments revealed that the antibodies had different characteristics. Whereas the antibody directed against the N-terminus of CT (SA2) did not influence CT/membrane interaction, the new antibody (SA209) against the alpha-helical domain of the enzyme interfered with this interaction. Our results provide experimental evidence that the alpha-helical domain (amino acid residues 228-287) of CT may serve as a membrane-interacting domain.

Animals↗
PubMed1994

Mutational analysis of human endothelin receptors ETA and ETB identification of regions involved in the selectivity for endothelin 3 or cyclo-(D-Trp-D-Asp-Pro-D-Val-Leu).

Two endothelin(ET)-receptor subtypes have been identified in mammals. They differ in their affinity towards the ET isopeptides with ETA displaying an ET-1-selective profile and ETB a non-selective one. To identify the regions responsible for the differential selectivity, chimeric forms were engineered by sequentially exchanging extracellular regions together with their flanking transmembrane domains. Two sets of reciprocal receptor mutants were thereby generated and analysed by expression in COS-7 cells. The recombinant receptor chimeras were characterised by direct and competitive radioligand-binding analysis. COS-7 cells transfected with vectors for the mutant receptors exhibited specific saturable [3-125I]iodotyrosyl ET-1 (125I-ET-1) binding, with affinities comparable to those of the wild-type receptors (apparent Ki approximately 1-6 x 10(-9) M). An average of 10(5)-10(6) binding sites/cell was calculated for the wild-type and mutant forms. In competition experiments using 125I-ET-1 and unlabeled ET-3, an ETB-selective agonist, we detected a clear switch from an ET-1-selective profile to a non-isopeptide-selective profile in ETA chimeras where the second extracellular loop and the flanking transmembrane domains IV and V, or the third extracellular loop and the flanking transmembrane domains VI and VII, had been exchanged for the corresponding parts of ETB. The opposite effect, namely a switch from a non-isopeptide-selective to an ET-1-selective binding, was observed for the mirror ETB chimeras where the symmetrical exchange had been operated. Using 125I-ET-1 and the ETA-specific antagonist cyclo-(D-Trp-D-Asp-Pro-D-Val-Leu) (BQ123), we were able to map the main determinants responsible for this selectivity to the N-terminal moiety of this receptor. Therefore, the ability for the interaction with ET-3 or BQ123 is governed by two different regions of the ET receptors.

Amino Acid Sequence↗
PubMed1994

Purification of human big endothelin 1 derived through cleavage with collagenase and dipeptidylpeptidase IV from a fusion protein expressed in Escherichia coli.

The cDNA coding for human big endothelin 1 (bigET-1), preceded by an optimized collagenase recognition sequence and followed by a stop codon, was fused in frame to the C-terminal region of alkaline phosphatase (AP). The fusion protein (AP-bigET), expressed in Escherichia coli K12 upon the lowering of organic phosphate concentrations, consisted of alkaline phosphatase (1-447), the collagenase cleavage site (Gly-Pro-Ala)4, and glycylprolyl-bigET-1. AP-bigET accumulated intracellularly in the form of inclusion bodies that were extensively washed and finally extracted by 8 M urea to yield highly enriched AP-bigET. Upon digestion of the fusion protein with collagenase, two disulfide conformeres of glycylprolyl-bigET-1 (bigET-1A and bigET-1B) could be purified by reverse-phase FPLC. Upon treatment with dipeptidylpeptidase IV to remove the N-terminal glycylprolyl-dipeptide, the later-eluting form of bigET-1 (bigET-1B) coeluted with authentic human bigET-1 on reverse-phase HPLC. BigET-1A and bigET-1B were formed at a ratio of 1:3. After reduction and S-pyridylethylation, both conformers coeluted with authentic but reduced bigET-1. Their amino acid sequences were identical. Both forms were converted by digestion with pepsin to the respective ET-1 conformeres (ET-1A and ET-1B) that were purified. In vasoconstriction assays, ET-1B but not ET-1A, at 10(-8) M, evoked a maximal response indistinguishable from that of authentic ET-1.

Alkaline Phosphatase↗
PubMed1994

The influence of diazepam on learning processes impaired by pentylenetetrazol kindling.

Repeated administration of pentylenetetrazol (PTZ) induces kindling and impairs shuttle-box learning. The available literature suggesting a close connection between seizure frequency and mental deficits in human epileptics allows us to hypothesize that seizure inhibition prevents the progressive mental retardation associated with kindling. In order to investigate the effect of motor seizure inhibition on mental impairment we administered diazepam (DZP) doses of 0.5 and 2.5 mg/kg, respectively 60 min prior to the 10 convulsant injections. After completion of kindling the learning performance of the rats was tested in the shuttle-box. PTZ kindling resulted in diminished shuttle-box learning. In control rats treated with DZP no significant changes in their learning ability occurred. Although DZP was found to suppress kindling development effectively a worsened shuttle-box learning could be observed in all PTZ groups treated with DZP.

Animals↗
PubMed1994

Effects of the peptide BCH-325 upon the efficacy of common antiepileptic drugs.

It was shown that BCH-325 (Pro-D-Phe-Pro-Gly), a des-tyrosine derivative of beta-casomorphin, exhibits anticonvulsive effects. In the present study, we combined the peptide with clinically used antiepileptic drugs (phenytoine, carbamazepine, diazepam, phenobarbital, and dipropyl acetate) to investigate possible interactions and, moreover, to draw conclusions concerning the mode of action of BCH-325. There were no interactions with phenytoine and carbamazepine, which might be interpreted to mean that BCH-325 exerted no action on sodium channels. The efficacy of the combinations with the other drugs (diazepam, phenobarbital, dipropyl acetate) was increased. The data suggest that the anticonvulsant effect of BCH-325 might occur by interference with elements of GABA/benzodiazepinergic neurotransmission.

Amino Acid Sequence↗
PubMed1994

Purification, cloning, and expression of a human enzyme with acyl coenzyme A: cholesterol acyltransferase activity, which is identical to liver carboxylesterase.

An enzyme with acyl coenzyme A:cholesterol acyltransferase (ACAT) activity was isolated from porcine liver, and sequences derived from trypsinized peptides indicated homology to liver carboxylesterase. By use of degenerate primers, human cDNA clones were identified, which were identical to human liver carboxylesterase. Expression of the full-length cDNA in Chinese hamster ovary (CHO) cells led to an approximately threefold increase in cellular ACAT activity. This was accompanied by an approximately 20-fold increase of cellular cholesteryl ester content. By light and electron microscopy, recombinant CHO cells contained numerous lipid droplets that were not present in control CHO cells. Expression of an antisense cDNA in HepG2 cells reduced cellular ACAT activity by 35% compared with control. To further investigate the role of the enzyme in cellular cholesterol homeostasis, regulation of the mRNA was investigated in 7-day cultured human mononuclear phagocytes (MNPs). When these cells were incubated in lipoprotein-deficient serum for 18 hours, the mRNA for ACAT/carboxylesterase was almost not detectable on Northern blots, whereas after incubation with acetylated low-density lipoproteins, a strong hybridization signal was obtained. This is evidence that the mRNA of ACAT/carboxylesterase is induced by cholesterol loading. It is concluded from the data presented that ACAT/carboxylesterase is relevant for cellular cholesterol esterification in vivo. The regulation in MNPs indicates that the enzyme is also involved in foam cell formation during early atherogenesis.

Amino Acid Sequence↗
PubMed1994

[Cytogenetic studies of Chilean children with acute lymphoblastic leukemia].

Acute lymphoblastic leukemia (ALL) is the most frequent childhood cancer. The leukemic cells of ALL patients show several well defined numeric and structural chromosomal abnormalities which are universally known for its prognostic implications. We studied a group of 44 children with ALL, to investigate the incidence of chromosome aberrations in ALL, its lymphocyte lineage and some clinical feature associations, and the finding of non previously described aberrations. A high proportion of patients (79.5%) showed chromosomal abnormalities. Most of them had a pseudodiploid karyotype (46 chromosomes), characterized mainly by a translocation. In relation to chromosome number, 27% of them were hyperdiploid with more than 50; 9% hyperdiploid between 47-50 and 7% hypodiploid (less than 46). Among structural aberrations found, were the following recurrent translocations: t (1; 19), t (4; 11), t (9; 22) in 6.8%, 9.1% and 2.3% of cases respectively, all related to an early B immunophenotype. Other translocations found, compromised regions 7q22, 9p21 - 24. Two new translocations in ALL were found: t (1; 5)(q23; q33), apparently balanced, and t (13; 21)(q14; q22), unbalanced. Other recurrent structural changes found were: deletion (6q), (7q), (9p), (11q), (12p), inversion (3q), isochromosome (7q), maker chromosomes and double minutes. The distribution of chromosome abnormalities in this group of patients was in agreement with previous reports from other investigators.

Adolescent↗
PubMed1994

[Anemia associated with acute post streptococcal glomerulonephritis].

Thirty-three children with post-streptococcal acute glomerulonephritis, age mean: 8.3 years (range: 6 - 12) were studied prospectively. Mean initial hematocrit (Hct) was 31.6% with 90% showing Hct under the normal lower limit for this age group. Reticulocyte index (RI) was < 0.5 in half of the cases. Serum iron concentration, total iron binding capacity (TIBC) and percentage of transferrin saturation were normal for this age group although 75% of the children had increased serum ferritin levels. At the time of discharge, Hct increased to 35.1% but 44% still had anemia. Hct increased spontaneously for 105 days stabilizing at 38%. Based on Hct changes, 3 groups were defined: Group I (3 individuals): normal upon discharge; Group II (19): partial recovery at discharge, slow recovery stabilizing after 105 days; Group III (11): lower Hct, slower recovery but with RI significantly higher than group II (0.96 vs 0.45 p < 0.01). Our data suggest that although hemodilution is present in all, it may be considered the solely factor only in 3 cases (Group I). In group II, evidence of bone marrow depression was indicated by the low RI. On the other hand, the intense anemia that could not be justified only by hemodilution and marrow depression in group III, suggests other pathogenic factors.

Acute Disease↗