Biomedical subjects

N A Taylor

Publications and source records attributed to N A Taylor.

At least 37 records · Page 2Linked to original sources

PubMed1996

An evaluation of the role of skin temperature during heat adaptation.

This project sought to evaluate the importance of skin temperature during heat acclimation, using an isothermal-strain model. Two groups of seven matched males, participated (1 h per day, 10 days) in one of two conditions: (i) temperate physical training (TEMP: 22.4 +/- 0.7 degrees C, relative humidity (r.h.) 41.0 +/- 0.9%); or (ii) combined physical training and heat acclimation (HEAT: 38.2 +/- 0.7 degrees C, r.h. 39.7 +/- 1.3%). Isothermal strain was induced in both groups by rapidly elevating rectal temperature by 1 degree C (cycling), then holding it constant by manipulating external work. Subjects completed two three-phase heat stress tests (39.8 +/- 0.1 degrees C, r.h. 38.6 +/- 1.2), consisting of 20 min rest, then 20 min cycling at each of 30% and 45% of peak power, before and after each regimen. While there was a difference of 4.2 degrees C in mean skin temperature between treatments, both regimens elicited a similar peripheral sudomotor increase, indicating a core temperature dependent adaptation. However, based on significant pre- vs. post-acclimation decreases in average auditory canal temperature (0.4 +/- 0.1 degree C), average forehead skin blood flow (26%), average perceived exertion (11%), and a 5% increase in average forehead sweat rate (0.1 +/- 0.04 mg cm-2 min-1), the HEAT regimen elicited a more complete acclimation. While elevation in core temperature is critical to acclimation, it also appears necessary to expose subjects to an external thermal stress. This observation has not been previously demonstrated under conditions of isothermal strain, and verifies the importance of skin temperature elevation in the acclimation process.

Acclimatization↗

PubMed1996

Whole-body hyperhydration in endurance-trained males determined using radionuclide dilution.

Despite evidence of hypervolemia following endurance training, there is little information regarding corresponding extravascular fluid volumes. Quantification of such volumes relies upon radionuclide dilution methods, previously hampered by the loss of plasma albumin. It was our purpose to measure human body-fluid distribution in eight endurance-trained males, using a simultaneous radionuclide dilution technique, incorporating radioiodinated serum fibronogen (RISF). Fluid distribution was measured on three occasions, using 2 microCi of RISF, 8 microCi of 51 Cr-labeled erythrocytes, and 20 microCi of Na82Br and 450 microCi of 3H2O; to measure PV, erythrocyte (RCV), extracellular (ECFV), and total-body water (TBW) volumes, respectively. Respective volume means, standard deviations, and coefficients of variation were: 46.6 (+/- 4.9; 8.44%), 33.3 (+/- 2.9; 3.89%), 258.1 (+/- 12.1; 4.93%), and 654.2 (+/- 13.4; 3.24%) ml.kg-1. The incorporation of RISF provided a reliable modification to previous methods, and revealed a body-fluid expansion in endurance-trained males. It was concluded that such subjects were hyperhydrated, possessing proportionately expanded fluid volumes throughout both intravascular and extravascular spaces. This was attributed to training history and accompanying reductions in adiposity.

Body Fluids↗

PubMed1995

Differences in pH optima and calcium requirements for maturation of the prohormone convertases PC2 and PC3 indicates different intracellular locations for these events.

PC2 and PC3, which is also known as PC1, are subtilisin-like proteases that are involved in the intracellular processing of prohormones and proneuropeptides. Both enzymes are synthesized as propolypeptides that undergo proteolytic maturation within the secretory pathway. An in vitro translation/translocation system from Xenopus egg extracts was used to investigate mechanisms in the maturation of pro-PC3 and pro-PC2. Pro-PC3 underwent rapid (t1/2 < 10 min) processing of the 88-kDa propolypeptide at the sequence RSKR83 to generate the 80-kDa active form of the enzyme. This processing was blocked when the active site aspartate was changed to asparagine, suggesting that an autocatalytic mechanism was involved. In this system, processing of pro-PC3 was optimal between pH 7.0 and 8.0 and was not dependent on additional calcium. These results are consistent with pro-PC3 maturation occurring at an early stage in the secretory pathway, possibly within the endoplasmic reticulum, where the pH would be close to neutral and the calcium concentration less than that observed in later compartments. Processing of pro-PC2 in the Xenopus egg extract was much slower than that of pro-PC3 (t1/2 = 8 h). It exhibited a pH optimum of 5.5-6.0 and was dependent on calcium (K0.5 = 2-4 mM). The enzymatic properties of pro-PC2 processing were similar to that of the mature enzyme. Further studies using mutant pro-PC2 constructs suggested that cleavage of pro-PC2 was catalyzed by the mature 68-kDa PC2 molecule. The results were consistent with pro-PC2 maturation occurring within a late compartment of the secretory pathway that contains a high calcium concentration and low pH.

Amino Acid Sequence↗

PubMed1995

The topography of eccrine sweating in humans during exercise.

The purpose of this study was to investigate the distribution of steady-state sweating rates (msw), during stressful exercise and heat exposures. Six men completed 42-min trials: 2-min rest and 40-min cycling at 40% peak power in 36.6 degrees C (relative humidity 46.0%). The msw was monitored using ventilated capsules at the forehead, and at three additional sites. Repeat trials allowed monitoring from eleven skin surfaces. Auditory canal temperature (Tac) and 11 skin temperatures were measured. After normalising msw to the forehead response within subjects, differences in Tac and onset time thresholds, and transient and steady-state msw were examined. The pooled, lower torso msw onset [mean 45.5 (SEM 42.0) s] preceded that of the head [mean 126.5 (SEM 34.8) s, P < 0.05], but was not significantly different from the legs [mean 66.6 (SEM 25.7) s], upper torso [mean 80.2 (SEM 36.8) s] or arms [mean 108.6 (SEM 31.2) s]. Transient msw did not differ among regions (P = 0.16). Mean, steady-state forehead msw [3.20 (SEM 0.51) mg.cm-2.min-1] was not significantly greater than the scapula, forearm, hand, stomach and lower back msw (in descending order), but was greater than the chest [1.6 (SEM 0.2)], upperarm [1.6 (SEM 0.2)], calf [1.5 (SEM 0.3)] and thigh msw [1.0 (SEM 0.2), P < 0.05 for all comparisons]. The results did not support the caudal-to-rostral sweat onset evident during supine, resting heat stress. Equivalent Tac sweat thresholds existed between sites, while steady-state state msw topography varied among subjects and was not dominated by central regions.

Adult↗

PubMed1995

Thermal tolerance following artificially induced polycythaemia.

Polycythaemia has been shown to improve physical performance, possibly due to increased arterial oxygen transport. Enhanced thermoregulatory function may also accompany this manipulation, since a greater proportion of the cardiac output becomes available for heat dissipation. We further examined this possibility in five trained men, who participated in three-phase heat stress trials (20 min rest, 20 min cycling at 30% peak power (Wpeak) and 20 min at 45% Wpeak at 38.3 (SEM 0.7) degrees C [relative humidity 41.4 (SEM 2.9)%]. Trials were performed during normocythaemia (control) and polycythaemia, obtained by reinfusion of autologous red blood cells and resulting in significant elevation of arterial oxygen transport. During the polycythaemic trials, the subjects demonstrated diminished thermal strain, as evidenced by a significant reduction in cardiac frequency (fc: 12 beats.min-1 lower throughout the test; P < 0.05), and reduced auditory canal temperatures (Tac) during the latter 20-min phase (P < 0.05). Forearm sweat onset was more rapid (363.0 compared to 1083.0 s; P < 0.05), and forearm sweat rate (msw) sensitivity was elevated from 1.80 to 2.91.mg.cm-2.min-1.degrees C-1 (P < 0.05). Forehead msw was depressed during the final 20 min, while forearm msw was greater during all test phases, averaging 0.94 and 1.20 mg.cm-2.min-1, respectively, over the 60 min. Skin blood flows for the upper back, upper arm and forearm were reduced (P < 0.05). Polycythaemia enhanced thermoregulation, through an elevation in forearm sweat sensitivity and msw, but not via increased cutaneous blood flow. These modifications occurred simultaneously with decreases in fc and Tac, resulting in greater thermal tolerance.

Adult↗

PubMed1995

Preferred room temperature of young vs aged males: the influence of thermal sensation, thermal comfort, and affect.

BACKGROUND: While research has demonstrated that aged persons may show impaired thermoregulatory control, we do not know whether dysthermia in the aged results from altered behavioral or autonomic responses. Consequently, we investigated age-related differences concerning the ability to regulate room temperature. METHODS: Two groups of matched healthy males (22.9 yr and 66.9 yr) were subjects of this study. After equilibration at 24 degrees C (rh 50%), chamber temperature controllers were set into cooling mode. Using a dual position switch, subjects adjusted this temperature when air temperature moved outside their preferred range. Switch operation resulted in maximal cooling or heating, without a steady state. Subjective ratings of thermal sensation, discomfort, and affect were provided at each activation. RESULTS: Both groups controlled temperature equivalently: 24.9 degrees C (+/- 1.3, young) and 24.5 degrees C (+/- 1.5, elderly; p > .05). At cold-induced change points, the skin temperatures of the calf, thigh, chest, and hand were significantly lower in the elderly subjects. During the heat-induced changes, chest, hand, upper arm, and mean skin temperatures were also lower (p < .05). At cold-induced change points, the elderly group felt colder (p < .05), were less uncomfortable (p < .05), and felt better than the young subjects (p < .05). During heat-induced changes, thermal sensation was equivalent, the elderly were more comfortable (p < .05), and felt better (p < .05). CONCLUSIONS: Assuming thermal discomforture drives behavior, it is possible that elderly people may require a more intense thermal stimulus to elicit the appropriate behavioral responses in the home. It is also possible that such stimuli will result in a greater heat flow, elevating the risk of dysthermia in the aged.

Adult↗

PubMed1995

Redistribution of body fluids during postural manipulations.

Inter-compartmental body-fluid distribution is contingent upon posture, exercise state and environmental temperature. This investigation aimed at quantifying the distribution of intra- and extravascular fluid volumes during postural manipulations. Fluid shifts were measured in eight males utilizing a simultaneous, radionuclide dilution technique, in which radioiodinated serum fibrinogen, radiochromated erythrocytes, radiobromine and tritiated water were used to measure plasma, red cell, extracellular and total body water volumes. Subjects were exposed to three postural changes [seated (control), supine and standing] for 30 min at an air temperature of 22.0 degrees C, with each posture separated by 30 min seated rest. Total body water content remained stable throughout postural changes (P = 0.842). Relative to seated volumes, BV increased by 89 mL when supine, and decreased by 406 mL while standing (P = 0.003), with such shifts being primarily a result of plasma movement (P = 0.011). Red cell volume changes were not significant. Vascular fluid lost during standing was filtered into the interstitial compartment (P = 0.014), with the extracellular and intracellular volumes remaining unaffected. (P = 0.271 and P = 0.800, respectively). These observations confirmed the influence of posture on inter-compartmental body-fluid distribution. The intravascular fluid loss when standing was caused by the filtration of plasma into the interstitium, while, during supine rest, intravascular volume increased, reflecting fluid flux from the interstitium to the circulation.

Adult↗

PubMed1995

Physiological assessment of the RNZAF constant wear immersion suit: laboratory and field trials.

Laboratory and field immersion trials were undertaken to determine the thermal protection afforded by a constant wear immersion suit (CWIS) in operation with the Royal New Zealand Air Force (RNZAF). Six males wore each of two ensembles during head-out laboratory immersions in 5.0 +/- 0.1 degree C (mean +/- SD) water for a maximum of 3 h. Ensembles 1 and 2 consisted of the CWIS in addition to minimal and maximal likely undergarment insulations, respectively. Open sea field trials (water temperature = 13.8 +/- 0.7 degree C; Beaufort wind state = 0-4; Sea state = 0-2) were conducted for a maximum of 2 h, with subjects wearing ensemble two and remaining strike aircrew apparel (ensemble three). Analysis of rectal temperature (Tre) changes permitted calculation of time to hypothermia (t35) and the survival estimate of 34 degrees C (t34). Mean (+/- SEM) t35 was 78 +/- 11 (n = 6), 187 +/- 20 (n = 5, p < 0.05) and 98 +/- 5 min (n = 3) for ensembles one, two and three, respectively. Mean t34 was 96 +/- 15, 259 +/- 31 (p < 0.05), and 119 +/- 5 min, respectively. Immersed insulations of ensembles one and two were 0.035 +/- 0.002 and 0.150 +/- 0.015 degree C.m2.W-1, respectively. Thus, the difference between minimal and maximal operational insulation was a 4.3-fold increase in insulation, which facilitated a 2.7-fold increase in mean t34. The thermal protection afforded by the CWIS during field trials was not sufficient to ensure survival for the 12-h expected rescue time.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1995

Forehead skin temperature and thermal sensation during exercise in cool and thermoneutral environments.

BACKGROUND: The influence of forehead and mean skin temperature on thermal sensation during exercise in hot, cool, and thermoneutral environments is unclear. HYPOTHESIS: We hypothesized that forehead temperature, in contrast to other skin sites, would contribute significantly to thermal sensation during exercise in hot, cool, and thermoneutral environments. METHOD: Volunteer males (N = 14) performed 30 min of constant load exercise on a cycle ergometer during which thermal sensation, skin and rectal temperatures, and heart rate were collected. Each subject participated in a control (24 degrees C), hot (40 degrees C), and cool (8 degrees C) condition. RESULTS: Significantly higher mean skin temperatures occurred during exercise in the hot condition (M = 37.22 +/- 0.20 degrees C) compared to exercise in the neutral (M = 33.34 +/- 0.51 degrees C) and cool conditions (M = 27.92 +/- 0.22 degrees C). Forehead skin temperature in the neutral and cool conditions was significantly greater than forearm, hand, thigh, and calf skin temperatures (p < 0.05). In the hot condition, forehead temperature was significantly greater than back, chest, upper arm, and hand skin temperatures (p < 0.05). In contrast to the neutral and hot conditions, forehead skin temperature was significantly associated with thermal sensation during exercise in the cool condition (r consistently > 0.67). CONCLUSIONS: We conclude that forehead skin temperature, in contrast to other skin sites, contributed significantly to thermal sensation during exercise in the cool.

Adult↗

PubMed1994

Calcium- and pH-dependent aggregation and membrane association of the precursor of the prohormone convertase PC2.

PC2 is a subtilisin-like protease which is thought to be involved in the processing of prohormones and proneuropeptides in neuroendocrine cells. The mature 68-kDa enzyme is generated by intracellular proteolytic processing of a 75-kDa pro-PC2 polypeptide. Neuroendocrine cells contain at least two secretory pathways: the regulated pathway whereby secreted products are concentrated, stored in granules, and released in response to external stimulation of the cell, and the constitutive pathway, whereby secretory and plasma membrane proteins are continuously transported to the cell surface without prior concentration or storage. An important step in the sorting of proteins into these pathways is thought to involve the aggregation of proteins destined for storage granules. To define the mechanisms in the intracellular sorting of PC2 to secretory vesicles, the present study was undertaken to investigate the aggregation and membrane association properties of precursor and mature forms of PC2. Using material expressed in microinjected Xenopus oocytes, it was demonstrated that the 75-kDa pro-PC2 polypeptide undergoes calcium- and acid pH-dependent aggregation. Calcium exhibited an effect on the aggregation of pro-PC2 at pH 7.0 and 6.5, whereas below 6.5 aggregation was independent of calcium. Association of pro-PC2 with membranes was observed at pH 5.5, but not at pH 7.0, 6.5, nor 6.0. The mature 68-kDa PC2 polypeptide remained soluble under conditions that caused aggregation and membrane association of the 75-kDa propolypeptide. Deletion of COOH-terminal sequences had no effect on the association of pro-PC2 with membranes, whereas a peptide corresponding to amino acids 57-85 of the propeptide was able to partially compete the membrane associated pro-PC2 away from the membranes.

Amino Acid Sequence↗

PubMed1994

Autocatalytic maturation of the prohormone convertase PC2.

PC2 is a member of the eukaryotic family of subtilisin-like proteases, which is thought to participate in the processing of prohormones and proneuropeptides in neuroendocrine cells. PC2 is synthesized as a 69-kDa prepropolypeptide. The NH2-terminal signal sequence is removed during segregation within the endoplasmic reticulum, where glycosylation occurs to generate a 75-kDa propolypeptide. A combination of site-directed mutagenesis and a cell-free translation/translocation system from Xenopus eggs was used to investigate the processing of the pro-PC2 precursor. The 75-kDa polypeptide underwent slow cleavage after the sequence Arg-Lys-Lys-Arg84 to generate a 68-kDa mature enzyme. Cleavage was blocked when the tetrabasic sequence was deleted (PC2M3) or when the active site Asp142 was changed to Asn (PC2M4). This latter observation suggested that cleavage of the 75-kDa propolypeptide to the mature 68-kDa enzyme was autocatalytic. Incubation of the PC2M4 mutant with the wild type PC2 precursor resulted in cleavage of both the wild type polypeptide and the catalytically inactive PC2M4 mutant. This indicates that cleavage could occur through an intermolecular reaction. The results also demonstrate that the novel Xenopus egg extract translation/translocation system represents a powerful cell-free method for studying proteolytic processing of propolypeptides.

Amino Acid Sequence↗

PubMed1994

Attenuation of the cutaneous blood flow response during combined exercise and heat stress.

Skin blood flow (SkBF) was measured in six male subjects using laser-Doppler velocimetry, with zero-gradient auditory canal temperature (Tac) used as an index of body core temperature (Tc). Subjects performed incremental, upright cycling commencing at 40% peak power (Wpeak: 10 min), increasing every 4 min by 5% Wpeak thereafter. Trials were conducted in hot (ambient temperature (Ta) 36.7 +/- 0.2 degree C, relative humidity (rh) 46.1 +/- 3.2%; mean +/- S.D.), and neutral environments (Ta 19.6 +/- 0.3 degree C, rh 50.2 +/- 1.4%). SkBF increased with Tac in all subjects. Attenuation of SkBF occurred at the same Tac, relative SkBF and cardiac frequency (fc) between environments, but at a lower exercise intensity (40.8 +/- 0.8% versus 55.8 +/- 3.0% Wpeak) in the hot environment (p < 0.05). Data indicate that Tc thresholds for SkBF attenuation may exist. However, it is suggested that attenuation thresholds coincided with a reduced central blood volume, which may occur at a critical level of cutaneous blood pooling.

Adult↗

PubMed1994

The effects of exercise duration on dynamics of respiratory gas exchange, ventilation, and heart rate.

This study investigated the effect of exercise duration on the response dynamics of oxygen consumption (VO2), carbon dioxide output (VCO2), ventilation VE), and cardiac frequency (fc) following stepped changes in exercise intensity, by manipulating the duration of the pretransition exercise period. A group of 11 healthy men performed a stepped exercise intensity cycling protocol on three separate occasions, each consisting of a stepped increase from 55% to 65% peak oxygen consumption (VO2,peak) of 6-min duration, followed by a stepped decrease to 55% VO2,peak of 10-min duration. This stepped protocol was preceded by either 5, 15, or 60 min of cycling at 55% VO2,peak. The response times for each variable were calculated at 10% increments between the prestep baselines and poststep plateaux. Following the stepped increase, the response times for VO2 at the 50%, 60%, 70%, 80%, and 90% relative increments were significantly reduced in the 60-min condition compared to the 15-min condition (P < 0.05); however, the response times for VCO2, VE, and fc were not significantly altered across the three conditions. No significant differences were found in the response times for VO2, VCO2, VE, and fc, across the three conditions following the stepped decrease in exercise intensity. It was concluded that the faster response time of aerobic metabolism to a stepped increase in exercise intensity was mediated by increases in active muscle temperature, leading to improved oxygen utilisation.

Adult↗

PubMed1994

Skeletal muscle mechanics in osteoporotic and nonosteoporotic postmenopausal women.

The purpose of this study was to evaluate single-joint, dynamic muscle function of osteoporotic (OST) and nonosteoporotic (N-OST) women. Knee flexor and extensor function in postmenopausal women (6th decade OST, n = 15; 7th decade OST, n = 10; 6th decade N-OST, n = 6; 7th decade N-OST, n = 5) were evaluated at five angular velocities from 60 degrees.s-1 to 300 degrees.s-1. All subject groups had similar anthropometric measurements, but the 6th decade N-OST group were more physically active than the age-matched OST group. The OST and N-OST women produced peak torque at similar knee angles. The 6th decade N-OST women produced significantly greater knee extensor mean peak torque and angle specific torque, and mean work than any of the other three groups (P < 0.05). However, knee flexor function was equivalent throughout the groups for most comparisons, except those between the 6th decade N-OST and 7th decade OST. While previous research has shown an early loss of flexor muscle function in aging women, our data indicated that women with osteoporosis also experience a deterioration in quadriceps muscle function not encountered within the N-OST subjects. It is possible that such a change is precipitated by reduced physical activity, and may mirror deterioration in bone mineral content.

Aged↗

PubMed1994

Respiratory responses of elite oarsmen, former oarsmen, and highly trained non-rowers during rowing, cycling and running.

The position of the body and use of the respiratory muscles in the act of rowing may limit ventilation and thereby reduce maximal aerobic power relative to that achieved in cycling or running, in spite of the greater muscle mass involved in rowing. This hypothesis was investigated for three groups of male subjects: nine elite senior oarsmen, eight former senior oarsmen and eight highly trained athletes unskilled in rowing. The subjects performed graded exercise to maximal effort on a rowing ergometer, cycle ergometer and treadmill while respiratory minute volume (VE) and oxygen consumption (VO2) were monitored continuously. The VE at a given VO2 during intense submaximal exercise (greater than 75% of maximal VO2) was not significantly lower in rowing compared with that in cycling and treadmill running for any group, which would suggest that submaximal rowing does not restrict ventilation. At maximal effort, VO2, and VE for rowing were less than those for the other types of exercise in all the groups, although the differences were not statistically significant in the elite oarsmen. These data are consistent with a ventilatory limitation to maximal performance in rowing that may have been partly overcome by training in the elite oarsmen. Alternatively, a lower maximal VE in rowing might have been an effect rather than a cause of a lower maximal VO2, if maximal VO2 was limited by the lower rate of muscle activation in rowing.

Adult↗

PubMed1994

Epidemiology of hypothermia: fatalities and hospitalisations in New Zealand.

BACKGROUND: Hypothermia occurs within domestic and non-residential settings. Most epidemiological data originate from the northern hemisphere, with little data being generally available concerning cases from New Zealand and Australia. AIMS: The National Health Statistics Centre (New Zealand) records hospital discharges and deaths. This study isolated hypothermia cases, to quantify its incidence and identify risk groups. METHODS: The morbidity and mortality files for the years 1979-86 (cases = 3,808,717) and 1977-86 (cases = 259,325; respectively) were searched by three investigators. RESULTS: Hypothermia hospitalisations were identified (6.9 per 100,000 per year). There were 176 deaths from hypothermia, representing 0.07% of the 259,325 deaths from all causes for the same period (0.537 per 100,000 people per year); of these fatalities, 72.2% were classified as domestic, and 27.8% as non-residential; of the domestic fatalities, 86.6% were 65 + years and 35.5% of these were male. Within the non-residential category, 75.5% were aged 13-65, of which 94.6% were male. The hospitalisation incidence was 12.7 times the fatality incidence, with the majority of hospitalisations being of domestic origin (88.4% of total), and occurring mostly within the lower and upper age extremes. Neonatal domestic hypothermia accounted for 72.6% of all domestic hospitalisations, and the elderly constituted 72.0% of the remaining cases. The proportion of New Zealand fatalities caused by hypothermia was 0.067%; lower than reported in the United Kingdom. CONCLUSIONS: The two main non-neonatal groups contributing to cases of hypothermia were males aged 13-65 years, and the elderly. In the aged, the proportion of hypothermia-related deaths was no different from that associated with other disorders, however, the case-fatality ratio was three times greater, highlighting the need for improving prevention and management strategies.

Adolescent↗

PubMed1994

Identification of a point mutation in the thyrotropin receptor of the hyt/hyt hypothyroid mouse.

The hyt/hyt hypothyroid mouse has an autosomal recessive, fetal-onset, severe hypothyroidism related to TSH hyporesponsiveness and associated with elevated TSH. Our previous work has suggested that the hypothyroidism and TSH hyporesponsiveness may result from a mutation in the hyt/hyt TSH receptor (TSHr) of the thyroid gland. Based on DNA sequencing of the entire coding region of the TSHr gene from the wild-type BALB/cBY +/+ mouse, the +/+ TSHr is 92% and 94% identical at the nucleotide and amino acid residue levels, respectively, compared to the rat TSHr gene. The coding region of the hyt/hyt TSHr, compared to that of the +/+ TSHr, has a single base change, CCG to CTG, at nucleotide position 1666, which leads to the replacement of a highly conserved proline at amino acid position 556 with a leucine in transmembrane domain IV. This mutation was introduced by site-directed mutagenesis into the wild-type human TSHr and transiently expressed in COS-7 cells. Although the size and abundance of the mutant TSHr mRNA suggested that there was no effect on the nature of the mRNA, TSH binding and the response to TSH in transfected cells were abolished. Further studies are necessary to clarify how the Pro to Leu replacement interferes with receptor expression on the cell surface or influences TSH binding. These functional consequences of the mutation appear to account for the observed TSH hyporesponsiveness and hypothyroidism in the hyt/hyt mouse.

Amino Acid Sequence↗

PubMed1994

Insulin-releasing pituitary cells as a model for somatic cell gene therapy in diabetes mellitus.

Insulin delivery by somatic cell gene therapy was evaluated using murine pituitary AtT20MtIns-1.4 cells. These cells have been stably transfected to release human insulin by the introduction of a recombinant plasmid bearing a human preproinsulin cDNA under the control of a zinc-sensitive metallothionein promoter. 6 x 10(7) AtT20MtIns-1.4 cells were implanted subcutaneously into streptozotocin-diabetic mice immunosuppressed with cyclosporin A. Release of human insulin was assessed using a specific plasma human C-peptide assay. On days 1 and 2 after implantation human C-peptide concentrations were about 0.02 pmol/ml. Consumption of zinc sulphate solution (500 mg/l) as drinking fluid for days 3-5 increased plasma human C-peptide concentrations to 0.11 +/- 0.01 pmol/ml (mean +/- S.E.M.), n = 11, P < 0.01, and concentrations declined when zinc was discontinued. The extent of hyperglycaemia was slightly lower (P < 0.05) than in a group implanted with non-transfected AtT20 cells. The study was terminated after 9 days, and tumour-like aggregations of implanted cells were identified at autopsy. These comprised a large necrotic core with insulin-containing cells at the periphery. The study provides support for the view that somatic cell gene therapy offers a potential approach to insulin delivery in diabetes mellitus.

Animals↗