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H Macgregor

Publications and source records attributed to H Macgregor.

15 recordsLinked to original sources
PubMed2005

Correlation between in vitro aggregation and thromboxane A2 production in fresh, liquid-preserved, and cryopreserved human platelets: effect of agonists, pH, and plasma and saline resuspension.

BACKGROUND: Some of the tests used to assess the quality of fresh and preserved platelets (PLTs) include PLT number, PLT morphology, pH of the PLT medium, PLT response to hypotonic stress, and PLT aggregation to agonists. This study was performed to assess the function of fresh and preserved PLTs by their response to aggregation and their production of thromboxane A2 after in vitro stimulation with agonists. STUDY DESIGN AND METHODS: PLTs isolated by apheresis procedures were stored at 22 degrees C for as long as 5 days and then frozen with 6 percent dimethyl sulfoxide, stored at -80 degrees C, thawed, washed, and resuspended in medium. The effects of agonists and the pH and composition of the medium on PLT aggregation and PLT production of thromboxane A2 after stimulation were measured. RESULTS: The agonists and the pH and composition of the medium affected both the aggregation response and the production of thromboxane A2 by the fresh and preserved PLTs. PLT aggregation response to arachidonic acid (AA) and adenosine diphosphate (ADP) was significantly lower in the cryopreserved PLTs than in the fresh and preserved PLTs. After stimulation with AA and ADP, the cryopreserved PLTs produced more thromboxane than did the fresh and liquid-preserved PLTs. CONCLUSIONS: The agonists and the pH and composition of the medium affected the response to aggregate and produce thromboxane in vitro in both the fresh and the liquid-preserved PLTs. PLT thromboxane A2 production may be a better in vitro test than PLT aggregation to assess PLT function in vivo.

Adenosine Diphosphate↗
PubMed2004

In vitro testing of fresh and lyophilized reconstituted human and baboon platelets.

BACKGROUND: Studies have been performed on human fresh, liquid-preserved, and cryopreserved platelets (PLTs) to assess PLT-adhesive surface receptors, PLT membrane procoagulant activity, PLT aggregation, and thromboxane production. Lyophilization has been developed as a method to preserve PLTs. This study was performed to evaluate these measurements on human and baboon fresh and lyophilized reconstituted PLTs. STUDY DESIGN AND METHODS: In both human and baboon fresh and lyophilized PLTs, aggregation response and PLT production of thromboxane A2 were measured after stimulation, and PLT surface markers P-selectin, glycoprotein (GP) Ib, GPIIb-IIIa, and factor (F) V were measured before and after stimulation. RESULTS: Fresh PLTs responded to the dual agonists arachidonic acid and adenosine diphosphate (ADP) to aggregate and produce thromboxane A2, and in both the PLT surface markers P-selectin and GPIIb-IIIa increased and GPIb decreased after stimulation. Neither human nor baboon lyophilized reconstituted PLTs aggregated to dual agonists, and neither produced thromboxane A2, increased PLT surface markers P-selectin or GPIIb-IIIa, or decreased PLT GPIb after stimulation. Nevertheless, after recalcification the lyophilized reconstituted PLTs accumulated FV to a significantly greater degree than fresh PLTs. CONCLUSIONS: Lyophilized reconstituted PLTs exhibited modification of the PLT membrane that interfered with aggregation and thromboxane production, prevented increases in PLT P-selectin and GPIIb-IIIa and decreases in GPIb after stimulation, and increased FV accumulation after recalcification. The in vitro data suggest that lyophilized PLTs may have reduced in vivo survival. In vivo studies are needed to determine the survival and function of lyophilized PLTs.

Animals↗
PubMed2002

Circulation and distribution of autotransfused fresh, liquid-preserved and cryopreserved baboon platelets.

BACKGROUND AND OBJECTIVES: Studies were carried out in five healthy male baboons to determine the 111indium oxine (111In-oxine) survival of autologous fresh, liquid-preserved and cryopreserved platelets. Simultaneous organ-distribution studies were performed to determine the percentage uptake of platelets by the spleen and/or liver. MATERIALS AND METHODS: Each of five baboons was transfused, on three different occasions, with autologous fresh platelets stored at 22 degrees C for 18 h, liquid-preserved platelets stored at 22 degrees C for 5 days and washed previously frozen platelets, labelled with 111In-oxine. RESULTS: In vivo recovery at 2 h was 81% for the fresh platelets, 54% for the previously frozen platelets and 44% for the 5-day-old liquid-preserved platelets. The weighted mean life span was 5.4 days for fresh platelets, 4.2 days for previously frozen platelets and 2 days for liquid preserved platelets. Increased radioactivity was detected over the liver 2 h after transfusion for both the previously frozen and liquid-preserved platelets. CONCLUSIONS: Cryopreserved platelets and liquid-preserved platelets stored at 22 degrees C for 5 days had reduced survival 2 h post-transfusion and reduced life span values compared to fresh platelets. In addition, the finding of increased radioactivity over the liver in the baboons that received cryopreserved and liquid-preserved platelets suggested that the liver was the site for removal of the non-viable platelets.

Animals↗
PubMed2001

Transcripts of the MHM region on the chicken Z chromosome accumulate as non-coding RNA in the nucleus of female cells adjacent to the DMRT1 locus.

The male hypermethylated (MHM) region, located near the middle of the short arm of the Z chromosome of chickens, consists of approximately 210 tandem repeats of a BamHI 2.2-kb sequence unit. Cytosines of the CpG dinucleotides of this region are extensively methylated on the two Z chromosomes in the male but much less methylated on the single Z chromosome in the female. The state of methylation of the MHM region is established after fertilization by about the 1-day embryonic stage. The MHM region is transcribed only in the female from the particular strand into heterogeneous, high molecular-mass, non-coding RNA, which is accumulated at the site of transcription, adjacent to the DMRT1 locus, in the nucleus. The transcriptional silence of the MHM region in the male is most likely caused by the CpG methylation, since treatment of the male embryonic fibroblasts with 5-azacytidine results in hypo-methylation and active transcription of this region. In ZZW triploid chickens, MHM regions are hypomethylated and transcribed on the two Z chromosomes, whereas MHM regions are hypermethylated and transcriptionally inactive on the three Z chromosomes in ZZZ triploid chickens, suggesting a possible role of the W chromosome on the state of the MHM region.

Animals↗
PubMed1998

Specific chromomeres on the chicken W lampbrush chromosome contain specific repetitive DNA sequence families.

Chromomeres 1 and 3 of the chicken W lampbrush chromosome contain most of the EcoRI and XhoI repeat sequence families respectively. These chromomeres were stained with DAPI and their sizes relative to other W chromomeres were observed. Their relative contents of EcoRI and XhoI repeats were determined using fluorescence in situ hybridization with genomic probes for each of the two repeat families. There were two types of W chromosome in the chickens (White Leghorn and Rhode Island Red) used in this study with respect to the amount of EcoRI repeat. A high-copy-number type has about 4000 copies of the 1.2-kb repeat per genome and shows a large fluorescence signal on W chromomere 1. A low-copy-number type has about 700 copies per genome and does not have a detectable chromomere 1 on W chromosome, nor does it show FISH labelling in the region normally occupied by chromomere 1. The genome of Fayoumi chickens has about one-sixth the amount of the XhoI sequence family of White Leghorns. W lampbrush chromomere 3 is much smaller and its FISH labelling with the XhoI probe is much weaker in Fayoumis than in White Leghorns. These results demonstrate that in the chicken W chromosome, specific chomomeres are occupied by specific DNA repeat sequence families.

Animals↗
PubMed1997

Molecular characterization and cytological mapping of a non-repetitive DNA sequence region from the W chromosome of chicken and its use as a universal probe for sexing carinatae birds.

A non-repetitive genomic DNA region of about 25 kb was cloned from the W chromosome of chicken using a genomic library prepared from a single W chromosome of the chicken. This region was mapped by fluorescence in situ hybridization (FISH) with mitotic and lampbrush chromosomes to a position between the major EcoRI family and the pericentromeric Xhol family on the W chromosome. A 0.6-kb EcoRI fragment (EE0.6) subcloned from this region consists of a sequence that can be obtained by the exon-trapping procedure and flanking sequences. Sequences, which are closely similar to that of EE0.6, are widely conserved on the W chromosomes of Carinatae birds, as revealed by Southern blot hybridization to HindIII-digested female and male genomic DNAs from 18 species of birds belonging to eight different taxonomic orders. The female sex of those birds can be determined by the presence of an unambiguous female-specific band. For many species of birds, the female sex can also be determined by polymerase chain reaction (PCR) using a set of primers from the flanking sequences in the chicken EE0.6.

Animals↗
PubMed1996

Characterization of DNA sequences constituting the terminal heterochromatin of the chicken Z chromosome.

Two clones, pCZTH5-8 and pCZTH12-8, were isolated from a female chicken genomic library by screening with sequences obtained from genomic libraries which had been constructed from a terminal region of a single Z chromosome of chicken utilizing laser microbeam irradiation and PCR amplification. Fluorescence in situ hybridization to the mitotic Z chromosome and the lampbrush ZW bivalent of chicken demonstrated that both the cloned sequences are located in the heterochromatic region of the Z chromosome at the end opposite to the pairing region with the W chromosome. The sequences pCZTH-8 and pCZTH12-8 are distributed widely on both the telomeric bow-like loops (TBL) and the region I (short loops region) of the Z lampbrush chromosome. These clones, pCZTH12-8 particularly notably, hybridized also to the TBLs of lampbrush bivalents 1-4 of chicken. Both sequences are transcribed in the lampbrush stage oocytes on the Z chromosome and on other macrobivalents. The subfragment of pCZTH5-8 which hybridizes to the TBLs and the insert of pCZTH12-8 contain regions that are closely similar in sequence. The pCZTH-8 sequence has no internal repeats and may be part of the 24-kb macrosatellite repeating unit that is evident after Nhel digestion of the genomic DNA. A cloned 24-kb unit, pFN-1, does not show significant DNA curvature, but cytosines of its CpG dinucleotides may be highly methylated in vivo. This contrasts with the repeat sequences of the W heterochromatin which not only have highly methylated CpG but are also strongly curved. The 24-kb unit is repeated about 830 times in the diploid genome of a female chicken, suggesting that nearly the entire terminal heterochromatin on the Z chromosome consists of this macrosatellite family. Sequences of the greater part of the pCZTH-8 are restricted to the genus Gallus but the sequence of one subregion which hybridizes to TBLs is present in the genomes of the order Galliformes.

Animals↗
PubMed1995

Single stranded nucleic acid binding structures on chicken lampbrush chromosomes.

In chicken oocytes, proteins of the K/J family or their analogs, such as are known to be involved in mRNA processing in humans, are closely associated with nascent C-rich RNA transcripts on the loops of lampbrush chromosomes. Using labelled single stranded nucleotide probes and an antibody to protein K, these C-rich transcripts have been mapped to six different pairs of lampbrush loops situated on 3 macrochromosomes, the sex bivalent (ZW) and certain microchromosomes. Each of these loop pairs has a distinctive morphology. The observations represent cytological evidence of the connection between K-proteins and C-rich RNA transcripts. Another structure, the spaghetti marker of macrochromosome II, also preferentially binds C-rich homonucleotides. This spaghetti marker has a highly distinctive fine structural organization that is quite unlike that of lampbrush loops. Its proteins are not recognised by antibodies to protein K. Homonucleotide binding loops are recommended as potentially extremely valuable as markers on physical maps of chicken chromosomes.

Animals↗
PubMed1993

Avian sex chromosomes in the lampbrush form: the ZW lampbrush bivalents from six species of bird.

The ZW bivalent has been identified and characterized in detail in its lampbrush form in oocytes of chicken, quail, turkey, pigeon, chaffinch and sparrow. The sex bivalent in all six species looks like a single highly asymmetrical chromosome. Most of it has the typical lampbrush organization. The terminal one-fifth is relatively thick and condensed and bears only a few pairs of lateral loops: this condensed terminal region is the W chromosome; the part with normal lampbrush morphology is the Z. The two are connected by a single near terminal chiasma. The fine scale morphology and arrangement of loops and markers on Z and W chromosomes are described for each species and lampbrush maps have been constructed. The identification of the lampbrush sex bivalent is based on the following criteria. The asymmetrical chromosome has two centromere regions. In the interstitial region of the asymmetrical chromosome where the junction between Z and W chromosomes is supposed to be, there are telomere-specific loops and telomeric DNA sequences and there is good morphological evidence for the presence of a chiasma. There are W chromosome specific DNA sequences in the region of the asymmetrical lampbrush chromosome that is thought to represent the W. Breed-specific variations in the morphology of the chicken W chromosome with respect to the sizes, numbers and arrangements of axial chromomeres and distributions of specified repeated DNA sequence families have been identified, offering one of the first examples of definitive correlation between a repeat family and a single chromomere. The lampbrush chromosomes of all the birds examined, except quail, terminate in distinctive free hanging loops. These are a novel feature in the sense that at the end of each chromatid there is a large transcription unit terminating in a cluster of telomeric DNA sequences.

Animals↗
PubMed1992

A novel structure associated with a lampbrush chromosome in the chicken, Gallus domesticus.

At a site near the end of the short arms of lampbrush bivalent 2 in the chicken (Gallus domesticus) there is always a marker structure that appears in the phase-contrast light microscope as a solid object with diffuse edges measuring about 4 microns across. When examined by transmission electron microscopy in thin section, this object appears as a loose bundle of fibres. In some preparations individual fibres appear 15-16 nm thick, smooth in outline and solid in cross-section. In other preparations they are 32-38 nm thick, rougher in outline and ring-like in cross-section. High-resolution scanning electron micrographs of the chromosome 2 marker show it to be a loose bundle of spaghetti-like fibres that is quite unlike anything previously seen on a lampbrush chromosome of any organism. As with the sectioned material, fibres in some preparations were smooth and 15-16 nm in diameter, whereas those in others were more knobbly and about 35 nm thick. The fibres appear to branch and in some cases it is clear that the daughter strands of a branch have the same dimensions as the parent strand. Free ends are rare. Total length of fibre material present at one marker locus is estimated to be between 500 and 2000 microns. Similar structures are not present on the lampbrush chromosomes of quail, wood pigeon or chaffinch. The nature of this fibrous marker, referred to in this paper as the "spaghetti marker", is discussed in relation to lampbrush chromosome function and to events that take place during the lampbrush phase of oogenesis in chicken. Evidence is discussed in relation to the possibility that the chromosome 2 marker represents a novel form of nuclear RNP or the specific association of some structural protein with one chromosome locus.

Animals↗
PubMed1979

The nucleolus organizers of Plethodon and Aneides located by in situ nucleic acid hybridization with Xenopus 3H-ribosomal RNA.

The main clusters of ribosomal genes, or nucleolus organizers, have been located by in situ nucleic acid hybridization of Xenopus laevis 3H labelled ribosomal RNA to mitotic chromosomes in squash preparations of intestinal epithelium from 7 species of Plethodon and 3 species of Aneides. The species used were chosen on account of having well known karyotypes and genome sizes. The Plethodon species covered a range of genome size of 20--69.4 pg. The locations of those nucleolus organizers that could be detected by autoradiography after in situ hybridization varied from species to species, and in Aneides there were differences between two populations of the same species. On the other hand, some distantly related species of Plethodon, with widely different genome sizes, had nucleolus organizers at corresponding positions. The results are discussed in relation to ideas on karyotype stability, homosequentiality and chromosome repatterning.

Animals↗