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Biomedical subjects

D A Thompson

Publications and source records attributed to D A Thompson.

At least 91 records · Page 5Linked to original sources
PubMed1997

Identification of a gene (GPR30) with homology to the G-protein-coupled receptor superfamily associated with estrogen receptor expression in breast cancer.

Using the technique of differential cDNA library screening, a cDNA clone was isolated from an estrogen receptor (ER)-positive breast carcinoma cell line (MCF7) cDNA library based upon the overexpression of this gene compared to an ER-negative cell line (MDA-MB-231). Sequence analysis of this clone determined that it shared significant homology to G-protein-coupled receptors. This receptor, GPCR-Br, was abundantly expressed in the ER-positive breast carcinoma cell lines MCF7, T-47D, and MDA-MB-361. Expression was absent or minimal in the ER-negative breast carcinoma cell lines BT-20, MDA-MB-231, and HBL-100. GPCR-Br was ubiquitously expressed in human tissues examined but was most abundant in placenta. GPCR-Br expression was examined in 11 primary breast carcinomas. GPCR-Br was detected in all 4 ER-positive tumors and only 1 of 7 ER-negative tumors. Based upon PCR analysis in hybrid cell lines, the gene for GPCR-Br (HGMW-approved symbol GPR30) was mapped to chromosome 7p22. The pattern of expression of GPCR-Br indicates that this receptor may be involved in physiologic responses specific to hormonally responsive tissues.

Amino Acid Sequence↗
PubMed1997

Chemically synthesized SDF-1alpha analogue, N33A, is a potent chemotactic agent for CXCR4/Fusin/LESTR-expressing human leukocytes.

Stromal cell-derived factor (SDF) 1 is a potent chemoattractant for leukocytes through activation of the receptor CXCR4/Fusin/LESTR, which is a fusion co-factor for the entry of T lymphocytotropic human immunodeficiency virus type 1 (HIV-1). This CXCR4-mediated HIV-1 fusion can be inhibited by SDF-1. Because of its importance in the study of immunity and AIDS, large scale production of SDF-1 is desirable. In addition to recombinant technology, chemical synthesis provides means by which biologically active proteins can be produced not only in large quantity but also with a variety of designed modifications. In this study, we investigated the binding and function of an SDF-1alpha analogue, N33A, synthesized by a newly developed native chemical ligation approach. Radioiodinated N33A showed high affinity binding to human monocytes, T lymphocytes, as well as neutrophils, and competed equally well with native recombinant SDF-1alpha for binding sites on leukocytes. N33A also showed equally potent chemoattractant activity as native recombinant SDF-1alpha for human leukocytes. Further study with CXCR4/Fusin/LESTR transfected HEK 293 cells showed that N33A binds and induces directional migration of these cells in vitro. These results demonstrate that the chemically synthesized SDF-1alpha analogue, N33A, which can be produced rapidly in large quantity, possesses the same capacity as native SDF-1alpha to activate CXCR4-expressing cells and will provide a valuable agent for research on the host immune response and AIDS.

Binding, Competitive↗
PubMed1997

Disregulation of mitotic checkpoints and regulatory proteins following acute expression of SV40 large T antigen in diploid human cells.

SV40 large T antigen (T) inactivates the tumor suppressor proteins p53 and pRb, and can induce cells to enter DNA replication at inappropriate times. We show here that T also compromises three cell cycle checkpoints that regulate the entry into and exit from mitosis. Human diploid fibroblasts infected with a retrovirus expressing T displayed an attenuated radiation-induced mitotic delay, were more susceptible to chemical-induced uncoupling of mitosis from the completion of DNA replication, and were more likely to exit mitosis and rereplicate their DNA when mitotic spindle assembly was inhibited. Consistent with altered mitotic checkpoint control, cells expressing T displayed elevated protein levels and/or associated activities of the mitotic regulatory proteins cyclin A, cyclin B, Cdc25C and p34(cdc2). These changes in mitotic control were evident within 5-10 population doublings after retroviral infection, indicating a direct effect of T expression. Cells acutely infected with the T-expressing retrovirus suffered numerical and structural chromosome aberrations, including increases in aneuploidy, dicentric chromosomes, chromatid exchanges and chromosome breaks and gaps. These findings indicate that T rapidly disrupts mitotic checkpoints that help maintain genomic stability, and suggest mechanisms by which T induces chromosome aberrations and promotes the immortalization and neoplastic transformation of human cells.

Antigens, Viral, Tumor↗
PubMed1997

Neuropeptide Y receptor genes on human chromosome 4q31-q32 map to conserved linkage groups on mouse chromosomes 3 and 8.

Npy1r and Npy2r, the genes encoding mouse type 1 and type 2 neuropeptide Y receptors, have been mapped by interspecific backcross analysis. Previous studies have localized the human genes encoding these receptors to chromosome 4q31-q32. We have now assigned Npy1r and Npy2r to conserved linkage groups on mouse Chr 8 and Chr 3, respectively, which correspond to the distal region of human chromosome 4q. Using yeast artificial chromosomes, we have estimated the distance between the human genes to be approximately 6 cM. Although ancient tandem duplication events may account for some closely spaced G-protein-coupled receptor genes, the large genetic distance between the human type 1 and type 2 neuropeptide Y receptor genes raises questions about whether this mechanism accounts for their proximity.

Animals↗
PubMed1997

Identification of two estrogen receptor transcripts with novel 5' exons isolated from a MCF7 cDNA library.

Two novel transcripts of human estrogen receptor (ER) have been identified that differ in the 5' untranslated sequence. It has previously been determined that an alternate ER transcript is generated from transcription initiated upstream of the main ER cap site (P1), and utilizes a splice acceptor site at +163. Here we report the isolation of 21 ER clones from a MCF7 cDNA library. Eleven of these clones correspond to transcripts that initiate at the P1 cap site, whereas the remaining 10 clones are derived from two previously unidentified ER transcripts (designated E and H) that both utilize the +163 splice acceptor site. A panel of breast and endometrial carcinoma cell lines were screened by reverse transcriptase-polymerase chain reaction (RT-PCR) for expression of the E and H transcripts. It was found that all ER-positive cell lines expressed both of the novel transcripts. In addition, 10 primary human breast cancers were analyzed, of which six expressed the E transcript and five abundantly expressed the H transcript. These data indicate that expression of ER in human breast cancers can be dependent upon an alternate promoter at least 20 kb upstream of the primary cap site for ER.

Base Sequence↗
PubMed1997

Mutations in RPE65 cause autosomal recessive childhood-onset severe retinal dystrophy.

Autosomal recessive childhood-onset severe retinal dystrophy (arCSRD) designates a heterogeneous group of disorders affecting rod and cone photoreceptors simultaneously. The most severe cases are termed Leber congenital amaurosis (LCA), while the less aggressive forms are usually considered juvenile retinitis pigmentosa. Recently, mutations in the retinal-specific guanylate cyclase gene were found in patients with LCA. Disease genes implicated in other forms of arCSRD are expected to encode proteins present in the neuroretina or in the retinal pigment epithelium (RPE). The RPE, a monolayer of cells separating the vascular-rich choroid and the neuroretina, is in intimate contact with the outer segments of rods and cones via the microvilli surrounding the photoreceptors. The RPE expresses a tissue-specific and evolutionarily highly conserved 61 kD protein (RPE65) present at high levels in vivo. Although the function of RPE65 is not yet known, an important role in the RPE/photoreceptor vitamin-A cycle is suggested by the fact that RPE65 associates both with serum retinol-binding protein and with the RPE-specific 11-cis retinol dehydrogenase, an enzyme active in the synthesis of the visual pigment chromophore 11-cis retinal. Here we report that the analysis of RPE65 in a collection of about 100 unselected retinal-dystrophy patients of different ethnic origin revealed five that are likely to be pathogenic mutations, including a missense mutation (Pro363Thr), two point mutations affecting splicing (912 + 1G-->T and 65 + 5G-->A) and two small re-arrangements (ins144T and 831del8) on a total of nine alleles of five patients with arCSRD. In contrast to other genes whose defects have been implicated in degenerative retinopathies, RPE65 is the first disease gene in this group of inherited disorders that is expressed exclusively in the RPE, and may play a role in vitamin-A metabolism of the retina.

Age of Onset↗
PubMed1996

Characterization of the human type 2 neuropeptide Y receptor gene (NPY2R) and localization to the chromosome 4q region containing the type 1 neuropeptide Y receptor gene.

Neuropeptide Y (NPY) signals through a family of G-protein-coupled receptors present in the brain and sympathetic neurons. To further our understanding of the genetic elements involved in the regulation of NPY receptor expression, we have cloned and characterized the human gene encoding the type 2 NPY receptor (Y2 receptor, HGMW-approved symbol NPY2R).2 The transcript spans 9 kb of genomic sequence and is encoded on two exons. As in the type 1 NPY receptor (Y1 receptor) gene, the 5'-untranslated region of the Y2 receptor is interrupted by an intervening sequence ( approximately 4.5 kb). However, the Y2 receptor gene does not contain an intron analogous to that present in the coding region of the Y1 receptor. The predicted transcript size ( approximately 4.5 kb) is consistent with the size observed by Northern analysis. The 381-amino-acid sequence deduced from the open reading frame is identical to that encoded by the cDNA. The Y2 receptor gene maps to human chromosome 4q31, the same region containing the Y1 receptor locus, suggesting that these subtypes may have arisen by gene duplication despite their structural differences.

Amino Acid Sequence↗
PubMed1996

How accurate are waiting time perceptions of patients in the emergency department?

STUDY OBJECTIVE: To assess the ability of patients to accurately estimate specific waiting times in the emergency department. METHODS: A questionnaire was administered by telephone to a random sample of 776 patients (or parents or responsible caretakers, if appropriate) who had been treated within the previous 2 to 4 weeks in the ED of a suburban hospital. Respondents were asked their perceptions of two particular time frames: (1) the time elapsed from triage until initial examination by the emergency physician (physician waiting time [PWT]), and (2) the time elapsed from triage until departure from the ED (total waiting time [TWT]). Corresponding actual times were extracted from a computerized database. Time frames were divided into discrete periods for comparison. The correspondence between actual and perceived times was assessed by optimal data analysis. RESULTS: Only 22.3% of the respondents accurately estimated PWT. Although this level of accuracy is statistically significant (P < .0001), it reflects only 11% of the theoretically possible improvement in accuracy beyond chance. More respondents overestimated than underestimated PWT (49.9% versus 27.8%, respectively). In contrast, TWT was accurately estimated by 36.6% of the respondents (P < .0001), reflecting 18% of the theoretically possible improvement in accuracy beyond chance. Fewer respondents overestimated than underestimated TWT (24.5% versus 38.9%, respectively). CONCLUSION: Patients are not very accurate in their estimation of actual waiting times. Although fewer than one fourth of the respondents overestimated the TWT spent in the ED, almost half the respondents overestimated the PWT.

Emergency Service, Hospital↗
PubMed1996

Effects of actual waiting time, perceived waiting time, information delivery, and expressive quality on patient satisfaction in the emergency department.

STUDY OBJECTIVE: To determine the effects of actual waiting time, perception of waiting time, information delivery, and expressive quality on patient satisfaction. METHODS: During a 12-month study period, a questionnaire was administered by telephone to a random sample of patients who had presented to a suburban community hospital emergency department during the preceding 2 to 4 weeks. Respondents were asked several questions concerning waiting times (ie, time from triage until examination by the emergency physician and time from triage until discharge from the ED), information delivery (eg, explanations of procedures and delays), expressive quality (eg, courteousness, friendliness), and overall patient satisfaction. RESULTS: There were 1,631 respondents. The perception that waiting times were less than expected was associated with a positive overall satisfaction rating for the ED encounter (P < .001). Satisfaction with information delivery and with ED staff expressive quality were also positively associated with overall satisfaction during the ED encounter (P < .001). Actual waiting times were not predictive of overall patient satisfaction (P = NS). CONCLUSION: Perceptions regarding waiting time, information delivery, and expressive quality predict overall patient satisfaction, but actual waiting times do not. Providing information, projecting expressive quality, and managing waiting time perceptions and expectations may be a more effective strategy to achieve improved patient satisfaction in the ED than decreasing actual waiting time.

Adolescent↗
PubMed1996

The full moon and ED patient volumes: unearthing a myth.

To determine if there is any effect of the full moon on emergency department (ED) patient volume, ambulance runs, admissions, or admissions to a monitored unit, a retrospective analysis of the hospital electronic records of all patients seen in an ED during a 4-year period was conducted in an ED of a suburban community hospital. A full moon occurred 49 times during the study period. There were 150,999 patient visits to the ED during the study period, of which 34,649 patients arrived by ambulance. A total of 35,087 patients was admitted to the hospital and 11,278 patients were admitted to a monitored unit. No significant differences were found in total patient visits, ambulance runs, admissions to the hospital, or admissions to a monitored unit on days of the full moon. The occurrence of a full moon has no effect on ED patient volume, ambulance runs, admissions, or admissions to a monitored unit.

Ambulances↗
PubMed1996

Visual acuity in unilateral cataract.

BACKGROUND: Patching the fellow eye in infancy is a well recognised therapy to encourage visual development in the lensectomised eye in cases of unilateral congenital cataract. The possibility of iatrogenic deficits of the fellow eye was investigated by comparing the vision of these patients with untreated unilateral patients and binocularly normal controls. METHODS: Sweep visual evoked potentials (VEPs) offer a rapid and objective method for estimating grating acuity. Sweep VEPs were used to estimate acuity in 12 children aged between 4 and 16 years who had had a congenital cataract removed in the first 13 weeks of life. The acuities of aphakic and fellow phakic eye were compared with the monocular acuities of similarly aged children who have good binocular vision, and with children with severe untreated uniocular visual impairment. Recognition linear acuities were measured with a linear Bailey-Lovie logMAR chart and compared with the sweep VEP estimates. RESULTS: A significant difference was found between Bailey-Lovie acuity of the fellow eye of the patient group and the right eye of binocular controls, and the good eye of uniocular impaired patients (one way ANOVA, p < 0.01). However, this was not evident for a similar comparison with sweep VEP estimates. There was no significant difference between the right and left eye acuities in binocular controls measured by the two techniques (paired t test). CONCLUSION: A loss of recognition acuity in the fellow phakic eye of patients treated for unilateral congenital cataract has been demonstrated with a logMAR chart. This loss was not apparent in children who have severe untreated uniocular visual impairment and may therefore be an iatrogenic effect of occlusion. An acuity loss was not apparent in the patient group using the sweep VEP method. Sweep VEP techniques have a place for objectively studying acuity in infants and in those whose communication difficulties preclude other forms of behavioural test. The mean sweep VEP acuity for the control groups is 20 cpd--that is, about 6/9. When acuities higher than this are under investigation--for example, in older children, slower transient VEP recording may be more appropriate, because higher spatial frequency patterns are not as visible at higher temporal rates (for example, 8 Hz used in sweep VEP recordings).

Adolescent↗
PubMed1996

Inability to follow up ED patients by telephone: there must be 50 ways to leave your number.

OBJECTIVE: To determine the ability to complete follow-up home telephone calls to ED patients. METHODS: A retrospective review of the ability to reach 4,741 patients called during a posttreatment patient satisfaction survey of visits to a suburban community hospital ED. RESULTS: Only 54.9% of all patients called could be contacted after three or fewer phone calls. Of the 2,139 (45.1%) who could not be reached, there was no answer for 1,260 (58.9%), despite three telephone calls to a number currently in service. A significant minority (21.1%) had given nonworking numbers. Another 12.4% indicated that no one by the name of the patient lived at the number called and 2.9% of the respondents alleged that the patient was deceased. CONCLUSIONS: Almost half of the patients who present to the ED and supply home telephone numbers give telephone numbers at which they cannot be reached in follow-up. Using telephone follow-up alone to reach patients seen in the ED may be an unreliable method of communication.

Data Collection↗
PubMed1995

A novel putative neuropeptide receptor expressed in neural tissue, including sensory epithelia.

We have used a homology based approach to identify G protein-coupled receptors preferentially expressed in retinal and taste cells. Rat and bovine sequences encoding a novel G protein-coupled receptor have been isolated. Analysis indicates that while the protein sequence is most similar to the receptors for somatostatin and opiates, it is unlikely to be a subtype of these receptors. Northern and RNase protection analysis indicates that the gene is preferentially expressed in neural and sensory tissues.

Amino Acid Sequence↗
PubMed1995

The optokinetic response differences between congenital profound and nonprofound unilateral visual deprivation.

BACKGROUND: The occurrence of monocular naso-to-temporal optokinetic nystagmus (OKN) asymmetry, as a reflection of the immature oculomotor system in infants, and its persistence with early onset monocular visual deprivation, is well known. This asymmetry has been linked with poor binocular function and attributed to disruption of the development of binocular cortical projections to the pretectum. Optokinetic nystagmus symmetry in patients with congenital uniocular total pattern vision deprivation has not been fully investigated. METHODS: The authors compared the optokinetic responses in six children with "profound" uniocular visual deprivation, who were born with untreated conditions (microphthalmos and persistent hyperplastic primary vitreous), with ten aphakic children treated early for congenital unilateral cataracts (nonprofound unilateral visual deprivation). Eye movements were recorded using dc-electro-oculography, and OKN was elicited using a full-field patterned curtain. Flash and pattern visual-evoked responses were also measured in each subject. RESULTS: Latent nystagmus was present in six children in the nonprofound group, whereas none was detected in the profound group. All children in the nonprofound group showed statistically significant monocular naso-to-temporal asymmetry for either eye. Subjects in the profound group had symmetric OKN. CONCLUSIONS: The authors conclude that unequal input from the two eyes and interocular rivalry lead to OKN asymmetry. Their results suggest that if vision from one eye is so negligible that it does not compete with the neuroanatomic connections of the fellow eye, then the input from this eye remains undisturbed, and OKN remains symmetric.

Adolescent↗
PubMed1995

Molecular characterization of the human gene encoding an abundant 61 kDa protein specific to the retinal pigment epithelium.

The retinal pigment epithelium (RPE) of the eye expresses an abundant 61 kDa protein (RPE65), that is developmentally regulated and tissue-specific. In our efforts toward understanding the specialized functions and development of the RPE, and the origins of inherited retinal degenerations, we have characterized the human gene encoding the 61 kDa protein. This is the first structural characterization of a gene transcribed specifically in the RPE. The gene maps to human chromosome 1p31. The sequence encoding the transcript spans over 20 kb, and is interrupted by 13 introns. A putative transcription start site lies 54 bp upstream of the initiation codon. A single transcript of approximately 2.9 kb is present in human RPE, and is not detected in other tissues. The deduced 533 amino acid sequence of the human protein is 98.7% identical to the bovine, but shows no significant similarity to any other entry in the databases. Expression of the 61 kDa protein appears to depend on the presence of environmental cues, since the corresponding transcripts are rapidly lost from RPE cells established in culture. Down regulation may occur post-transcriptionally, since AU-rich elements proposed to target RNA for rapid degradation are present throughout the 3'-untranslated region. The tissue-specific expression, high abundance, evolutionary conservation, developmental regulation, and sequence of the 3'-untranslated region suggest that the 61 kDa protein is the product of a functionally important gene whose expression is tightly regulated.

Amino Acid Sequence↗
PubMed1995

Relating patient satisfaction to waiting time perceptions and expectations: the disconfirmation paradigm.

OBJECTIVE: To determine the association of patient satisfaction with waiting time perceptions and expectations. METHODS: A random sample of patients seen at one community hospital ED during a one-year period was surveyed by telephone within two to four weeks of evaluation to determine perceived satisfaction and waiting time perceptions and expectations. RESULTS: In response to 3,641 attempted phone surveys, 1,574 patients had usable interviews. Consistent with a hypothesis derived using the disconfirmation paradigm (i.e., that satisfaction is a function of the magnitude and direction of the difference between perceived service and expected service), the patients were least satisfied when waiting times were longer than expected, were relatively satisfied when waiting times were perceived as equal to expectations, and were highly satisfied when waiting times were shorter than expected (p < 0.0001). Overall, the measure of effect strength (values from 0 to 1) of perceived waiting time vs expectation on the patient satisfaction score was 0.32, indicating moderate association. CONCLUSIONS: The current study supports the validity of the disconfirmation paradigm in relating patient satisfaction to waiting time perceptions and expectations. Furthermore, it emphasizes that achieving satisfaction in a service encounter necessitates that perceived performance meet or exceed patient expectations.

Data Collection↗
PubMed1995

Modulation of amblyopia therapy following early surgery for unilateral congenital cataracts.

BACKGROUND: Stimulus deprivation amblyopia is the principal cause of visual impairment in infants with unilateral congenital cataract. Even if lensectomy is undertaken at an early age, intensive postoperative occlusion of the phakic eye is essential for the development of useful vision in the aphakic eye. Despite this, the optimum method of regulating occlusion therapy is uncertain. METHODS: Interocular acuity differences identified using clinical preferential looking techniques (Keeler cards) were used to regulate target levels of phakic eye occlusion in a prospective evaluation of 10 systemically, metabolically, and neurologically normal infants in whom dense unilateral cataract was diagnosed before 8 weeks of age, and operated upon by 10 weeks. Actual occlusion levels were recorded each day by parents in a diary. The development of preferential looking acuity in the phakic and aphakic eye were compared with prediction intervals derived from observations on 43 normal children. RESULTS: Aphakic eye preferential looking acuities were within the normal range at last review in all but one infant. Interocular acuity differences were < or = 0.5 octave in all children older than 1 year of age at last review, and > or = 1 octave in three of four children less than 1 year old at last review (Fisher exact p = 0.033). Phakic eye acuities were within the normal range in all infants at all visits. CONCLUSION: Within the first 2 years of life, normal preferential looking acuity may be achieved in both eyes of infants undergoing early surgery for unilateral congenital cataract if occlusion therapy is modulated according to interocular acuity differences quantified by clinical preferential looking techniques.

Amblyopia↗