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Biomedical subjects

M Norman

Publications and source records attributed to M Norman.

At least 73 records · Page 4Linked to original sources
PubMed1994

Crystals of HIV-1 reverse transcriptase diffracting to 2.2 A resolution.

Reverse transcriptase (RT) from the human immunodeficiency virus type 1 has been crystallized in four closely related forms, the best of which diffract X-rays to 2.2 A resolution. The RT was crystallized as a complex with a non-nucleoside inhibitor, either nevirapine or a nevirapine analogue. Crystals grew from 6% PEG 3400 buffered at pH 5. These were of space group P2(1)2(1)2(1) with unit cell parameters a = 147 A, b = 112 A, c = 79 A (form A), with one RT heterodimer in the asymmetric unit. Changes in unit cell parameters and degree of crystalline order were observed on soaking pregrown crystals in various solutions, giving three further sets of unit cells. These were a = 143 A, b = 112, A, c = 79 A (form B), a = 141 A, b = 111 A, c = 73 A (form C), a = 143 A, b = 117 A, c = 66.5 A (form D). The last two forms diffract X-rays to 2.2 A resolution. Structure determinations of these latter crystal forms of RT should give a detailed atomic model for this therapeutically important drug target.

Crystallography, X-Ray↗
PubMed1994

Fine structure mapping of the human X-linked hypophosphatemic rickets gene locus.

X-linked hypophosphatemic rickets (HYP) is an X-linked dominant disorder characterized by decreased renal tubular phosphate reabsorption and consequent hypophosphatemia. Renal cross-transplantation studies in Hyp mice indicate that the disorder is secondary to the elaboration of an as yet unidentified humoral factor. A full understanding of the pathophysiology of the disease and the nature of this factor will be facilitated by identification of the HYP gene. Efforts to isolate the HYP gene have been deterred by limited precision in the map of the Xp22.1 region and the consequent distance between DXS365 and DXS274, the previously discovered flanking markers for the HYP gene. To map the HYP region precisely, HYP family resources from two groups of investigators were combined, and several newly available microsatellite repeat probes were tested for linkage to HYP. Our data indicate that DXS365, DXS3424, DXS443, DXS1052, DXS274, and DXS1683 are tightly linked to the HYP gene and suggest a locus order of: Xtel-DXS315-(GLR/DXS43)-DXS257-(DXS443+ ++-DXS3424)-DXS365-HYP-DXS1683-DXS1052-DXS 274-(DXS41/DXS92)-DXS451-Xcen. The HYP gene is located in the 350- to 650-kilobase region between DXS365 and DXS1683. These results will provide a basis for the isolation of candidate genes from the region.

Base Sequence↗
PubMed1994

The effect of pre-fitting counselling on the outcome of hearing aid fittings.

The effect of pre-fitting counselling on the outcome of fittings of NHS behind-the-ear hearing aids to adult first-time users was investigated. Questionnaires and diaries were sent both before and after fitting to 48 subjects who were given pre-fitting counselling and 47 control subjects, all of whom were fitted with standard NHS hearing aids in Southampton or Bath between September 1989 and July 1991. The test and control groups had similar distributions of age, sex and hearing loss. Analysis of the data showed that the counselling had no significant effect on levels of satisfaction, aid usage or benefit; these outcome measures also showed no significant correlation with any of the personal characteristics or attitude factors which were studied.

Aged↗
PubMed1994

Day of injury CT scan as an index to pre-injury brain morphology.

This study compared the ventricle-to-brain ratio (VBR) of the day-of-injury (DOI) computerized tomogram (CT) in traumatic brain-injured (TBI) patients with post-injury (2 months or greater) magnetic resonance (MR) VBRs in the same patients and in medical control subjects. The DOI VBR did not differ significantly from the medical controls, but both (DOI and medical control VBR) differed significantly from post-injury VBR. Additionally, a case study is presented wherein MR imaging studies were obtained prior to TBI so that a direct comparison of pre-injury to DOI to post-injury changes could be made. In this case the pre-injury and DOI VBRs were within approximately 9% of each other. In contrast, the post-injury VBR demonstrated over a 100% increase in comparison to either the pre-injury or DOI scan. This case and another case are illustrated using three-dimensional image analysis to represent ventricular change over time. These cases, along with the similarity of the DOI VBR with the medical controls, suggests that the DOI VBR can be utilized as an estimate or index of pre-injury ventricle/brain morphology. This will permit the use of DOI CT data for within-subject designs in TBI research that examines the course of degenerative changes over time.

Adolescent↗
PubMed1993

Flanking markers define the X-linked hypophosphatemic rickets gene locus.

X-linked hypophosphatemic rickets (HYP) is an X-linked dominant disorder characterized by decreased renal tubular phosphate reabsorption and consequent hypophosphatemia. The defect in tubular phosphate reabsorption is probably secondary to an unidentified humoral factor. Identification of the humoral factor and a full understanding of the pathophysiology of the disease await the identification of the HYP gene. Previously we demonstrated that DXS257 and DXS41 are flanking markers for the HYP gene. Two markers, DXS365 and DXS274, are tightly linked to the HYP gene, but investigators have been unable to determine whether they are centromeric or telomeric to the disease gene. Since tightly linked flanking markers are necessary prerequisites to obtain the gene by positional cloning techniques, we sought to determine the relative positions of these markers to the HYP gene by expanding our data base for linkage studies. We also investigated a new polymorphic probe for linkage to HYP to construct a more detailed genetic map around the HYP locus. Our data indicate that the markers DXS365, DXS274, and DXS92 are tightly linked to the HYP locus and suggest a locus order of Xtel-(DXS444/DXS315)-DXS43-(DXS257/DXS3 65)-HYP-(DXS274/DXS41/DXS92)-DXS-451- DXS319-Xeen. These results will facilitate attempts further to localize and clone the HYP gene.

Chromosome Mapping↗
PubMed1993

Skin microcirculation in neonatal polycythaemia and effects of haemodilution. Interaction between haematocrit, vasomotor activity and perfusion.

The cutaneous microcirculation was investigated before and 2-4 h after haemodilution in 13 newborn infants with polycythaemia. Skin microvascular perfusion was related to the haematocrit and to the presence of rhythmical changes in blood flow, reflecting vasomotor activity. The microcirculation was studied with a laser Doppler fluxmeter in a superficial microvascular bed represented by the dorsal hand skin. In five subjects, it was possible to combine laser Doppler flux measurements with microscopic quantitations of blood cell velocity in single, nailfold capillaries. In neonates less than 12 h postnatal age (n = 6), microvascular perfusion was only one-third that of the equally polycythaemic, but older infants (n = 7, p < 0.01). The higher perfusion in the older neonates with polycythaemia was associated with rhythmical variations in blood flow (3-5 cycles/min). There was no difference in skin temperature, blood pressure or heart rate between the two age groups. After haemodilution, the younger infants had developed rhythmical blood flow variations with the same characteristics as in the older group, in which the flow pattern was unchanged. In association, the laser Doppler flux had increased 304% in the younger and 73% in the older group (median values, p < 0.05 versus pretreatment values). The post-treatment change in laser Doppler flux corresponded to an increase in nailfold capillary blood cell velocity from 0.08 (0.02-0.23) mm/s prior to haemodilution to 0.21 (0.07-0.32) mm/s after treatment (n = 5, p < 0.05). Skin microcirculatory effects of neonatal polycythaemia and haemodilution vary in relation to vasomotor activity.

Aging↗
PubMed1993

Frequency analysis of the contralateral suppression of evoked otoacoustic emissions by narrow-band noise.

The influence of noise bandwidth on the contralateral masking of click-evoked otoacoustic emissions was investigated by recording emissions from 20 normal subjects with contralateral narrow-band noise, with wide-band noise and without contralateral noise for comparison. Narrow-band noise of five different centre frequencies and three different bandwidths per frequency was used at intensities of 40 and 60 dB SL. Analysis of the emissions showed that the 40-dB SL contralateral noise did not produce a significant amount of suppression. The suppression produced by 60-dB SL noise was spread throughout the frequency range of the emissions, with limited frequency specificity. There was some evidence that the amount of suppression increases with the bandwidth of the noise, particularly for noises centred on 1 and 2 kHz; the wide-band noise produced much greater suppression than any of the narrow bands. There was a large amount of inter-subject variation, which could not be explained by differences in the absolute level of the emissions. One subject with a total unilateral hearing loss in the contralateral ear was also tested, and gave no significant suppression.

Acoustic Stimulation↗
PubMed1993

Prostaglandin E2-induced ripening of the human cervix involves changes in proteoglycan metabolism.

OBJECTIVE: To elucidate how prostaglandin E2 (PGE2) induces cervical ripening. METHODS: Cervical biopsies were obtained immediately postpartum from women successfully treated with PGE2 gel intracervically. Six specimens were incubated with [35S]sulfate and five were used to characterize the nonlabeled proteoglycan composition. In separate experiments, biopsy specimens from three term pregnant women with unripe cervices were incubated with PGE2 in organ cultures. Proteoglycans were isolated and characterized using ion-exchange and gel chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. RESULTS: During PGE2-induced cervical ripening, the synthesis of proteoglycans, especially a large chondroitin/dermatan sulfate proteoglycan and biglycan, increased three- to sixfold. This resulted in a net increase in the large proteoglycan in the PGE2-treated cervices. In organ culture, on the contrary, incubation with PGE2 decreased the proteoglycan synthesis. CONCLUSION: Prostaglandin E2-induced cervical ripening is accomplished by increased remodeling of the cervical connective tissue, involving changed proteoglycan metabolism and composition.

Adult↗
PubMed1993

Changed proteoglycan metabolism in human cervix immediately after spontaneous vaginal delivery.

OBJECTIVE: To investigate whether the final cervical ripening just before or during labor corresponds to changes in cervical connective tissue metabolism and composition. METHODS: Cervical biopsies taken transvaginally immediately after normal vaginal delivery and at hysterectomy were incubated with [35S] sulfate. In other biopsy specimens, the nonlabeled proteoglycan pattern was studied. Collagen concentration and organization were monitored to estimate the degree of cervical ripeness. Labeled and nonlabeled proteoglycans were isolated, characterized, and quantitated. Collagen concentration and collagen, extractable and not extractable by pepsin digestion, were quantitated as hydroxyproline. RESULTS: A marked increase in the accumulation rate of labeled proteoglycans was observed in cervical biopsies obtained immediately after vaginal delivery. This increase was markedly larger than that previously noted in term pregnant women with unfavorable cervices. In particular, the accumulation of one labeled large chondroitin sulfate proteoglycan and one small dermatan sulfate proteoglycan, biglycan, increased significantly compared with the unfavorable cervices. In contrast, the small dermatan sulfate proteoglycan, decorin, showed a relative decrease. The changed synthesis of proteoglycans resulted in a two- to threefold increase in chemical amounts of the large proteoglycan, while the small dermatan sulfate proteoglycans decreased to approximately half of that in the nonpregnant cervix. CONCLUSION: The changed cervical proteoglycan metabolism results in a changed proteoglycan composition of the tissue. This alteration probably influences collagen organization, contributing to the cervical softening and dilatation essential for an uncomplicated vaginal delivery.

Adult↗
PubMed1992

An evaluation of skin capillary blood flow determinations in neonates using a computerized videophotometric method.

Dynamic capillaroscopy has been shown to give valid measurements of skin capillary blood flow in adults. The aim of this investigation is to evaluate the accuracy of the method when applied to studies in newborn infants. A computerized videophotometric technique was used to measure the skin capillary blood cell velocity (CBV) and the capillary erythrocyte column diameter at 64 standardized locations in 16 nailfold capillaries in five subjects. To estimate the method's reliability, we calculated the coefficient of variation (CV = 1 SD/mean x 100%) for repeated measurements at each location. The CV for CBV determinations was found to be 3.4% (0.9-13%) and the corresponding value for capillary diameter measurements was 6.7% (0.5-12%) (median and range). Although the CBV and diameter values were found to vary along the course of the capillary, the volume flow of blood cells was assumed to be the same in simultaneously studied but different sections of the capillary. The paired CBV and diameter observations were therefore used to calculate blood cell flow values at an average of four different intracapillary locations. To estimate the method's validity, the CV for these repeated flow calculations was determined and found to be 7.5% (3.2-26%). We conclude that the age-specific characteristics in neonates, mainly a high hematocrit and short, irregular skin capillary loops, do not limit the feasibility and accuracy of videophotometric microscopy for dynamic microcirculatory studies in neonates.

Capillaries↗
PubMed1992

Effects of neonatal polycythemia and hemodilution on capillary perfusion.

The effects of neonatal polycythemia on nutritive capillary perfusion were investigated by a television microscopy technique. The capillary blood flow velocity in skin was measured in 12 neonates with polycythemia before and after treatment with hemodilution, and in 13 healthy control infants. The capillary blood flow velocity in the patients was 0.11 (0.02 to 0.34) mm/sec and in the healthy control infants 0.30 (0.17 to 0.44) mm/sec (p less than 0.01, median and range values). In relation to the absolute hematocrit change after treatment range, -20% to 0%), the capillary blood flow velocity increased nonlinearly (range, +733% to -14%; r = -0.98; p less than 0.001). The postnatal age was found to contribute significantly to the variation in results--the neonates with polycythemia studied during the first day of life had a very slow skin capillary circulation and responded to treatment with a more pronounced increase in capillary blood flow velocity than did the older patients. This in vivo model for capillary perfusion indicates that an insufficient microcirculation may be involved in the pathophysiology responsible for the morbidity associated with neonatal polycythemia.

Age Factors↗
PubMed1992

Alanine-scanning mutagenesis of human prolactin: importance of the 58-74 region for bioactivity.

We have generated 10 alanine mutants of human PRL (hPRL), a member of the PRL/GH family, to investigate the involvement of the highly conserved 58-74 region in the biological behavior of the protein. When treated with polyclonal anti-hPRL antibodies, all mutants were immunologically indistinguishable from the unmodified hPRL, and circular dichroism analyses indicated that their alpha-helix content was similar to that of the unmodified hormone. Mutations C58A, K69A, and, to a lesser extent, P66A affected drastically the ability of hPRL first to bind to the lactogenic receptor and second to stimulate the proliferation of Nb2 lymphoma cells, proving the importance of the 58-74 peptide segment for hPRL bioactivity. Binding affinities of these mutants to the Nb2 lactogenic receptor have been compared to lactogenic binding data previously obtained for several mutants of hGH. The comparison reveals that the residues involved in the biological properties of the two proteins are not at topologically equivalent positions. Hence, we suggest that the binding of these hormones to the lactogenic receptors occurs through a different molecular mechanism having distinct requirements at the residue level.

Alanine↗
PubMed1992

Growth hormone binding protein in patients with renal failure.

To investigate changes in the growth hormone binding protein (GH-BP) in renal disease, gel chromatography was used to separate free and bound hormone after incubation with 125I-GH, the results being expressed as a percentage of radioactive GH eluting in a high molecular weight (70-80 kD) peak. In 26 normal individuals, binding was 39.3 +/- 8.0%, while in 11 patients with renal disease who were off dialysis binding was reduced to 16.8 +/- 5.6%. Similarly, in 9 patients undergoing hemodialysis binding was reduced to 24.6 +/- 6.8%, in 8 patients undergoing chronic ambulatory peritoneal dialysis binding was reduced to 25.7 +/- 7.6%, and in 9 patients within three months of a renal transplant binding was reduced to 25.1 +/- 8.6%. Scatchard analysis showed that these changes were not a result of decreased affinity of GH-BP for GH, and receptor binding studies showed that uremic serum was not inhibiting binding. The decreased concentration of GH-BP may indicate decreased expression of the GH receptor in target tissues, and hence diminished responsiveness to GH in renal failure.

Adult↗
PubMed1992

Preinductive cervical ripening with prostaglandin E2 in women with one previous cesarean section.

Thirty term pregnant women with one previous cesarean section and with unripe cervices were given 0.5 mg prostaglandin E2 in gel strictly intracervically for cervical ripening and labor induction. Fifteen out of these 30 women (50%) were vaginally delivered within 24 hours. The cervical ripening/labor induction was considered a failure in two women. In one woman, an episode of hypercontractility was registered. After tocolytic therapy the uterine activity was normalised and the woman had a normal vaginal delivery. The frequency of cesarean sections was 8/30 (27%). At the operations no insufficiencies in the uterine scars were noted. Conclusively, strict intracervical application of 0.5 mg prostaglandin E2 in gel can be used for cervical ripening and labor induction also in women with one previous cesarean section.

Administration, Intravaginal↗
PubMed1991

Proteoglycan metabolism in the connective tissue of pregnant and non-pregnant human cervix. An in vitro study.

Profound changes occur in the cervix during pregnancy. In particular, the connective tissue is remodelled. To elucidate the mechanisms behind this process, the metabolism of cervical connective tissue was studied using tissue cultures. Cervical biopsies from non-pregnant and pregnant women were incubated with [35S]sulphate. The proteoglycans of the tissue specimens were purified by ion-exchange and gel chromatography and characterized by SDS/PAGE and by enzymic degradation. In the non-pregnant cervix, the incorporation of [35S]sulphate into the proteoglycans was linear for 48 h. During the first 6 h of incubation the accumulation of chiefly one small labelled proteoglycan (apparent Mr 110,000) substituted with dermatan sulphate was recorded. This is in accordance with the known proteoglycan composition of non-pregnant cervical tissue. In addition, small amounts of two larger radioactive dermatan/chondroitin sulphate proteoglycans (apparent Mr values 220,000 and greater than 500,000) were recorded. After longer periods of incubation the proportion of heparan sulphate proteoglycans increased considerably. The pregnant tissue showed a clearly different composition of labelled proteoglycans. An increased accumulation of the two larger dermatan/chondroitin sulphate proteoglycans was seen in addition to the dominant small dermatan sulphate proteoglycan of the non-pregnant cervix. The rate of accumulation of these two proteoglycans was about 3 times higher in the pregnant tissue, whereas that of the small dermatan sulphate proteoglycan was only increased 2-fold. The fact that the concentration of proteoglycans in the pregnant cervix is approximately one-half of that in the non-pregnant cervix indicates that the turnover of proteoglycans in pregnant cervical tissue is significantly increased. The major effect of this profound change of metabolism was a 50% decrease in proteoglycan content and a 2-fold increased proportion of a dermatan sulphate proteoglycan with an apparent Mr of 220,000.

Adult↗
PubMed1991

Reactive hyperemia in term neonates and adults--a laser Doppler fluxmetry study of skin microcirculation.

The reactivities of neonatal and adult microcirculation have been studied and compared. The cutaneous reactive hyperemia after 1 and 4 min of arterial occlusion (AO) was measured with a laser-Doppler fluxmeter in 21 healthy neonates and 10 adults. Local skin temperature, mean arterial blood pressure (MAP), and skin prick hematocrit were also determined at the same time. The magnitude of neonatal reactive hyperemia was approximately one-third that of the adult response regardless of the duration of AO. In both groups, with age-specific regressions, the hyperemic blood flow response after 4 min of AO developed more slowly in subjects with low MAP and was of low magnitude in subjects with high hematocrit values. In response to a prolongation of AO, from 1 to 4 min, the magnitude and duration of hyperemia increased significantly and similarly in both neonates and adults. We conclude that compared to adults neonates have a less pronounced ability to increase skin microcirculation in response to local ischemia. The normally low blood pressure and high hematocrit in newborn infants contribute further to this conclusion.

Adult↗
PubMed1991

Levels of GH binding activity, IGFBP-1, insulin, blood glucose and cortisol in intensive care patients.

OBJECTIVE: To investigate levels of serum GH binding activity, insulin-like growth factor binding protein-1 (IGFBP-1), blood glucose, serum insulin, and cortisol in patients on the Intensive Therapy Unit. DESIGN: Case-control study of severely ill patients admitted to the Intensive Therapy Unit. PATIENTS: Six critically ill patients (51-78 years) who required ventilatory and nutritional support and six healthy age, sex, height and weight matched controls. MEASUREMENTS: Patients and controls were studied for two 24-hour periods; the patients before and after commencing parenteral nutrition, the controls whilst fasted and on a second occasion when fed a diet equal in protein and calories to that of the patients' parenteral nutrition. Samples were taken hourly for measurement of IGFBP-1, blood glucose, serum insulin and cortisol. Growth hormone binding activity was measured at 0 hours. RESULTS: Blood glucose levels were higher in the patients than controls in both the fasted (mean +/- SEM 5.1 +/- 0.5 vs 3.8 +/- 0.2 mmol/l, P = 0.04) and fed states (10.1 +/- 1.6 vs 5.0 +/- 0.1 mmol/l, P = 0.02) and patients' insulin levels were also higher when fed (81.5 +/- 31.6 vs 24.2 +/- 4.8 mU/l, P = 0.046) although there were no significant differences between patients and controls when fasted. IGFBP-1 levels were inversely related to insulin levels in both the patients and controls; mean IGFBP-1 concentrations were higher in fasted patients than in controls (123 +/- 38 vs 52 +/- 9, P = 0.046) but when fed, both groups had similar mean levels. Serum GH binding activity was low in the patients and did not change with feeding. Mean 24-hour cortisol levels were higher in the patients than in controls, whether fasted or fed, and showed no nyctohemeral rhythm. CONCLUSIONS: We have previously reported that critically ill patients have low levels of IGF-I with augmented basal levels of GH. The present results demonstrate that these changes in the GH-IGF-I axis are associated with insulin resistance with respect to blood glucose and high levels of IGFBP-1 when patients are fasted. However, when fed, the inverse relationship of IGFBP-1 to insulin is preserved. Patients have low levels of GH binding activity and increased mean cortisol levels. Interventional studies in this patient group with GH and IGF-I must take account of these changes in binding protein and cortisol levels.

Abdomen↗