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Biomedical subjects

A Harper

Publications and source records attributed to A Harper.

At least 37 records · Page 2Linked to original sources
PubMed2000

Epigenotype-phenotype correlations in Beckwith-Wiedemann syndrome.

Beckwith-Wiedemann syndrome (BWS) is a model imprinting disorder resulting from mutations or epigenetic events involving imprinted genes at chromosome 11p15.5. Thus, germline mutations in CDKN1C, uniparental disomy (UPD), and loss of imprinting of IGF2 and other imprinted genes have been implicated. Many familial BWS cases have germline CDKN1C mutations. However, most BWS cases are sporadic and UPD or putative imprinting errors predominate in this group. We have identified previously a subgroup of sporadic cases with loss of imprinting (LOI) of IGF2 and epigenetic silencing of H19 proposed to be caused by a defect in a distal 11p15.5 imprinting control element (designated BWSIC1). However, many sporadic BWS patients show biallelic IGF2 expression in the presence of normal H19 methylation and expression patterns. This and other evidence suggested the existence of a further imprinting control element (BWSIC2) at 11p15. 5. Recently, we showed that a subgroup of BWS patients have loss of methylation (LOM) at a differentially methylated region (KvDMR1) within the KCNQ1 gene centromeric to the IGF2 and H19 genes. We have now analysed a large series of sporadic cases to define the frequency and phenotypic correlates of epigenetic abnormalities in BWS. LOM at KvDMR1 was detected by Southern analysis or a novel PCR based method in 35 of 69 (51%) sporadic BWS without UPD. LOM at KvDMR1 was often, but not invariably associated with LOI of IGF2. KvDMR1 LOM was not detected in BWS patients with putative BWSIC1 defects and cases with KvDMR1 LOM (that is, putative BWSIC2 defects) invariably had a normal H19 methylation pattern. The incidence of exomphalos in putative BWSIC2 defect patients was not significantly different from that in patients with germline CDKN1C mutations (20/29 and 13/15 respectively), but was significantly greater than that in patients with putative BWSIC1 defects (0/5, p=0.007) and UPD (0/22, p<0.0001). These findings are consistent with the hypothesis that LOM of KvDMR1 (BWSIC2 defect) results in epigenetic silencing of CDKN1C and variable LOI of IGF2. BWS patients with embryonal tumours have UPD or a BWSIC1 defect but not LOM of KvDMR1. This study has further shown how (1) variations in phenotypic expression of BWS may be linked to specific molecular subgroups and (2) molecular analysis of BWS can provide insights into mechanisms of imprinting regulation.

Beckwith-Wiedemann Syndrome↗
PubMed1999

The World Health Organization WHOQOL-100: tests of the universality of Quality of Life in 15 different cultural groups worldwide.

The World Health Organization Quality of Life assessment (WHOQOL-100) was developed simultaneously across 15 international field centers and includes 24 facets relating to quality of life, which are grouped into 4 larger domains: physical, psychological, social relationships, and environment. It also includes 1 facet examining overall quality of life and general health perceptions. This article examines the extent to which the WHOQOL-100 assesses quality of life perceptions in different cultures and whether it is structurally comparable in these cultures. Regression analysis showed all 4 domains to be important in assessing quality of life in each of the 15 centers. Structural equation modeling suggested further support for the proposal that there are universal facets and domains that are cross-culturally important in determining quality of life and suggested that the ordering of facets within domains is comparable across cultures.

Adult↗
PubMed1999

Antioxidant effects of isoflavonoids and lignans, and protection against DNA oxidation.

The antioxidant capability of a series of isoflavonoid and lignan compounds in both cellular and cell-free systems has been investigated, and related to structure. Nordihydroguaiaretic acid exhibited a potent antioxidant activity in both HepG2 and MDA-MB-468 cells (IC50 5.3 and 1.1 microM respectively), as determined by inhibition of 2',7'-dichlorofluorescin oxidation via t-BOOH, although no inhibition was observed with other compounds tested in this system. All compounds inhibited the formation of 8-oxodeoxyguanosine in DNA exposed to hydroxyl radicals via gamma irradiation or the Fenton reaction. Whilst almost complete inhibition of gamma irradiation-induced damage was achieved (IC50 ranged from 0.2 to 0.8 microM), inhibition was less pronounced with the Fenton system. The ability of all compounds to interact with DNA (as well as with reactive oxygen and iron) was also demonstrated by scanning UV spectroscopy, suggesting that the compounds may inhibit DNA oxidation at least in part by binding to DNA. Hydroxyl radical-scavenging, iron-chelating and DNA-binding activity of these compounds supports their proposed role as natural cancer-protective agents.

Antioxidants↗
PubMed1998

Structure of d(TGCGCA)2 and a comparison to other DNA hexamers.

The X-ray crystal structure of d(TGCGCA)2 has been determined at 120 K to a resolution of 1.3 A. Hexamer duplexes, in the Z-DNA conformation, pack in an arrangement similar to the 'pure spermine form' [Egli et al. (1991). Biochemistry, 30, 11388-11402] but with significantly different cell dimensions. The phosphate backbone exists in two equally populated discrete conformations at one nucleotide step, around phosphate 11. The structure contains two ordered cobalt hexammine molecules which have roles in stabilization of both the Z-DNA conformation of the duplex and in crystal packing. A comparison of d(TGCGCA)2 with other Z-DNA hexamer structures available in the Nucleic Acid Database illustrates the elusive nature of crystal packing. A review of the interactions with the metal cations Na+, Mg2+ and Co3+ reveals a relatively small proportion of phosphate binding and that close contacts between metal ions are common. A prediction of the water structure is compared with the observed pattern in the reported structure.

Binding Sites↗
PubMed1997

Evaluation of beta-globin gene therapy constructs in single copy transgenic mice.

Effective gene therapy constructs based on retrovirus or adeno-associated virus vectors will require regulatory elements that direct expression of genes transduced at single copy. Most beta-globin constructs designed for therapy of beta-thalassemias are regulated by the 5'HS2 component of the locus control region (LCR). Here we show that a human beta-globin gene flanked by two small 5'HS2 core elements or flanked by a 5'HS3 (footprints 1-3) core and a 5'HS2 core are not reproducibly expressed in single copy transgenic mice. In addition, low copy transgene concatamers that contain only dimer 5'HS2 cores fail to express, whereas those that contain monomer 5'HS2 cores express at 14% per copy. These data suggest that spacing between HS cores is crucial for LCR activity. We therefore constructed a novel 3.0 kb LCR cassette in which the 5'HS2, 5'HS3 and 5'HS4 cores are each separated by approximately 700 bp. When linked to the 815 bp beta-globin promoter this LCR directs 45% levels of expression from four independent single copy transgenes. However, the 3.0 kb LCR linked to the 265 bp promoter expresses variable levels, averaging 18%, from three single copy transgenes. Our findings suggest that sequences in the distal promoter play a role in single copy transgene activation and that larger LCR and promoter elements are most suitable for gene therapy applications.

Animals↗
PubMed1997

Ictal cerebral perfusion related to EEG in drug resistant focal epilepsy of childhood.

OBJECTIVES: To evaluate the EEG changes during seizures in children with drug resistant focal epilepsy who demonstrate hypoperfusion at the "seizure focus" interictally, but no perfusion change during the seizure. METHODS: Ictal EEG findings of six children with focal epilepsy who demonstrated hypoperfusion on rCBF SPECT after an interictal injection of (99)Tc(m) HMPAO concordant with the seizure focus, but who did not demonstrate rCBF change after an ictal injection (group 1) were reviewed. These were contrasted with the EEG data of six children matched as closely as possible for age, type of epilepsy, and pathology who did show hyperperfusion at the seizure focus on ictal scan when compared with the interictal study (group 2). RESULTS: The children in group 1 showed slowing of the EEG at the time of the (99)Tc(m) HMPAO injection relative to that seen at the onset of the seizure. Those in group 2 showed rhythmic activity, or sharp waves, or both on EEG at the time of injection. This last change was also seen consistently when the EEG data of a further 13 children who also showed ictal hyperperfusion at the seizure focus were reviewed. CONCLUSION: Ictal rCBF does not invariably increase at the seizure focus in patients with drug resistant focal epilepsy.

Adolescent↗
PubMed1997

The effect of transmission bandwidth on diagnostic accuracy in remote fetal ultrasound scanning.

Six subspecialists with considerable experience in fetal ultrasound viewed a selection of pre-recorded ultrasound scans. Scans from 18 patients recorded on VHS video-tape were supplied from five centres in the UK and Ireland, each made on a high-resolution ultrasound machine by an experienced sonologist at a referral centre. Each observer viewed the scans on a large display monitor in an individual viewing booth. The scans were viewed in random order, at randomly selected bandwidths. Observers, who were blinded to both recording and bandwidth, assessed the technical quality on a five-point Likert scale. They also recorded their diagnosis. The six observers each carried out 32 viewing sessions, which gave a total of 192 viewings. There was no significant difference in the perceived technical quality of the scans between the two bandwidths used (P = 0.09). Of the 84 recordings transmitted at 1920 kbit/s, 71 (85%) were diagnosed correctly or 'half correctly' and 13 (15%) were misdiagnosed. Of the 95 recordings transmitted at 384 kbit/s, 66 (69%) were diagnosed correctly or 'half correctly' and 29 (31%) were misdiagnosed. This difference was significant (P = 0.03). The results indicate that although there were no perceived differences in technical quality between recordings transmitted at 384 or 1920 kbit/s, diagnostic accuracy was marginally worse at the lower bandwidth. This suggests that the higher bandwidth conveys more detail and information to the observer, which in turn enables more accurate diagnosis. However, further work is required before a definitive choice can be made about the optimum transmission bandwidth for remote fetal ultrasound studies.

Female↗
PubMed1996

The p53 response to ionising radiation in adult and developing murine tissues.

The induction of the p53 response to ionising radiation has been studied during murine development and in the adult animal. The response has been assessed by precise quantitative assay of p53 protein levels in tissues and by immunohistochemistry. Newly developed transgenic mice in which a lacZ transgene is driven by a p53 response element have also been used to directly assess the transcriptional activity of the induced protein. There is striking developmental control of the p53 response so that in early development all tissues accumulate high levels of p53 following radiation and indeed p53 is present at elevated levels in some unirradiated tissues. Later in development clear heterogeneity of the p53 response becomes apparent, both in terms of the responses of individual tissues and of cell populations, within those tissues. The study of lacZ transgene expression and the occurrence of apoptosis in different tissues that accumulate p53 protein point to a further level of control regulating the nature and degree of the downstream response to elevated levels of p53 in cells. These findings have important implications for the susceptibility of different tissue types to carcinogenic and other insults. The early expression of the p53 response is consistent with novel models of p53 function that suggest it may have evolved principally as a defense against teratogenic insult that permits plasticity of development.

Age Factors↗
PubMed1996

Exon 1 of the HD gene with an expanded CAG repeat is sufficient to cause a progressive neurological phenotype in transgenic mice.

Huntington's disease (HD) is one of an increasing number of neurodegenerative disorders caused by a CAG/polyglutamine repeat expansion. Mice have been generated that are transgenic for the 5' end of the human HD gene carrying (CAG)115-(CAG)150 repeat expansions. In three lines, the transgene is ubiquitously expressed at both mRNA and protein level. Transgenic mice exhibit a progressive neurological phenotype that exhibits many of the features of HD, including choreiform-like movements, involuntary stereotypic movements, tremor, and epileptic seizures, as well as nonmovement disorder components. This transgenic model will greatly assist in an eventual understanding of the molecular pathology of HD and may open the way to the testing of intervention strategies.

Animals↗
PubMed1996

A dominant chromatin-opening activity in 5' hypersensitive site 3 of the human beta-globin locus control region.

Single-copy human beta-globin transgenes are very susceptible to suppression by position effects of surrounding closed chromatin. However, these position effects are overcome by a 20 kbp DNA fragment containing the locus control region (LCR). Here we show that the 6.5 kbp microlocus LCR cassette reproducibly directs full expression from independent single-copy beta-globin transgenes. By testing individual DNase I-hypersensitive sites (HS) present in the microlocus cassette, we demonstrate that the 1.5 kbp 5'HS2 enhancer fragment does not direct beta-globin expression from single-copy transgenes. In contrast, the 1.9 kbp 5'HS3 fragment directs beta-globin expression in five independent single-copy transgenic mouse lines. Moreover, the 5'HS3 core element and beta-globin proximal promoter sequences are DNase I hypersensitive in fetal liver nuclei of these expressing transgenic lines. Taken together, these results demonstrate that LCR activity is the culmination of at least two separable functions including: (i) a novel activity located in 5'HS3 that dominantly opens and remodels chromatin structure; and (ii) a recessive enhancer activity residing in 5'HS2. We postulate that the different elements of the LCR form a 'holocomplex' that interacts with the individual globin genes.

Animals↗
PubMed1996

The human beta-globin locus control region confers an early embryonic erythroid-specific expression pattern to a basic promoter driving the bacterial lacZ gene.

The beta-globin locus control region (LCR) is contained on a 20 kb DNA fragment and is characterized by the presence of five DNaseI hypersensitive sites in erythroid cells, termed 5'HS1-5. A fully active 6.5 kb version of the LCR, called the muLCR, has been described. Expression of the beta-like globin genes is absolutely dependent on the presence of the LCR. The developmental expression pattern of the genes in the cluster is achieved through competition of the promoters for the activating function of the LCR. Transgenic mice experiments suggest that subtle changes in the transcription factor environment lead to the successive silencing of the embryonic epsilon-globin and fetal gamma-globin promoters, resulting in the almost exclusive transcription of the beta-globin gene in adult erythropoiesis. In this paper, we have asked the question whether the LCR and its individual hypersensitive sites 5'HS1-4 can activate a basic promoter in the absence of any other globin sequences. We have employed a minimal promoter derived from the mouse Hsp68 gene driving the bacterial beta-galactosidase (lacZ) gene. The results show that the muLCR and 5'HS3 direct erythroid-specific, embryonic expression of this construct, while 5'HS1, 5'HS2 and 5'HS4 are inactive at any stage of development. Expression of the muLCR and 5'HS3 transgenes is repressed during fetal stages of development. The transgenes are in an inactive chromatin conformation and the lacZ gene is not transcribed, as shown by in situ hybridization. These data are compatible with the hypothesis that the LCR requires the presence of an active promoter to adopt an open chromatin conformation and with models proposing progressive heterochromatization during embryogenesis. The results suggest that the presence of a beta-globin gene is required for LCR function as conditions become more stringent during development.

Animals↗
PubMed1995

Immunocytochemical localization of aspartate and glutamate in the peripheral vestibular system.

Controversy exists concerning the identity of the neurotransmitter in the mammalian peripheral vestibular system. Several candidates have been proposed, including the excitatory amino acids glutamate and aspartate and the inhibitory amino acid gamma-aminobutyric acid (GABA). Previous studies have demonstrated vestibuloneural electrophysiological activity associated with glutamate and aspartate. Paraffin sections of rat vestibular ganglia and end-organs were examined for the presence of glutamate-like and aspartate-like immunoreactivity. Our results demonstrate the presence of both aspartate-like and glutamate-like immunoreactivity in vestibular hair cells, peripheral vestibular nerve fibers, and vestibular ganglion cells. Minimal immunoreactivity was noted in the tissues surrounding these cells. These data add support to the hypothesis that the excitatory amino acids glutamate and aspartate are involved in vestibular neurotransmission.

Absorption↗
PubMed1995

Patterns of visual loss associated with pituitary macroadenomas.

PURPOSE: To describe the patterns of visual loss associated with pituitary macroadenomas compressing the anterior visual pathway. METHOD: A prospective survey of 29 patients with pituitary macroadenomas who presented to the neurosurgical unit at St Vincent's Hospital, Melbourne. Selected patients had histologically verified pituitary macroadenomas, and visual defects (acuity, colour and/or field loss) consistent with anterior visual pathway compression. RESULTS: All patients had visual field defects detected on perimetry, and the majority were asymmetrical. Bitemporal defects were most common but field defects ranged from monocular defects to generalised constriction. Four patients (13.8%) did not report visual symptoms, and of those who had symptoms, blurred vision was the most common complaint. Ninety-six per cent of eyes had field loss, 56% had decreased colour vision, 46% had decreased acuity, 31% had optic disc pallor, and 2% had an ophthalmoplegia. CONCLUSIONS: Patients with visual pathway compression by pituitary macroadenomas may be asymptomatic despite having field defects. Perimetry is the most sensitive method of identifying compression, followed by colour vision, visual acuity, then the presence of optic atrophy. Automated static threshold perimetry appears to show early field defects better than manual kinetic perimetry. All patients with pituitary macroadenomas should have thorough ophthalmological examinations, including perimetry to document visual deficits secondary to compression.

Adenoma↗
PubMed1995

The effect of temperature on spontaneous action potential discharge of the isolated sinus venosus from winter and summer plaice (Pleuronectes platessa)

The spontaneous cardiac pacemaker activity and conformation were recorded in vitro, using intracellular recording methods, from heart tissue of summer- and winter-caught plaice. The effects of changing temperature on the pacemaker rate, duration of action potential and diastolic depolarization were investigated by altering the temperature of the superfusing medium. The resting intrinsic rate of discharge was significantly greater in pacemaker cells from winter plaice (P=0.05), but there was no significant difference between winter and summer fish in the apparent Arrhenius activation energies for this process. However, there was a significant difference in the estimated intercept, indicating a thermal shift in the processes underlying the spontaneous pacemaker rhythm. There was no significant difference in the diastolic depolarization duration recorded from winter and summer fish over the temperature range 4&shy;22 &deg;C. The major effect of previous environmental temperature was on the duration of the action potential (P<0.02), indicating that the observed changes in pacemaker discharge rate were not influenced by the processes that determine the duration of the pacemaker diastolic depolarisation but were modulated by the channel events that give rise to the action potential.

Journal Article↗
PubMed1995

Menorrhagia management options.

A prospective study of the management of menorrhagia in new patients presenting to gynaecological outpatients was undertaken at four centres in Northern Ireland and two in Great Britain. 325 patients were enrolled, the majority of whom (87%) had severe menorrhagia. Patients in all six centres were similar in relation to age, marital status, parity, use of contraception and severity of symptoms. 62% of the patients were managed medically, improved and were discharged. The rates of surgical intervention, in particular in women aged less than 40, appeared higher in the Northern Ireland hospitals than Great Britain. There is a need to review and audit current practices in the management of menorrhagia.

Adolescent↗