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Biomedical subjects

D C Mitchell

Publications and source records attributed to D C Mitchell.

At least 19 recordsLinked to original sources
PubMed1996

Primary alcohols modulate the activation of the G protein-coupled receptor rhodopsin by a lipid-mediated mechanism.

The visual pigment rhodopsin is a prototypical member of the G protein-coupled receptor superfamily. In this study, we have investigated the effect of a series of n-alcohols on the formation of metarhodopsin II (MII), the photoactivated conformation of rhodopsin, which binds and activates transducin. When rhodopsin was photolyzed in the presence of several n-alcohols, increased MII formation was observed in the order ethanol > butanol > hexanol, whereas longer chain n-alcohols inhibited MII formation with decanol > octanol. The magnitude of the stimulatory effects was greater in a more highly unsaturated phospholipid. Alcohols, which enhanced MII formation also increased phospholipid acyl chain packing free volume, while those that decreased this bilayer property inhibited MII formation. An apparent discontinuity in the effect of these alcohols results when their potency is calculated in terms of the total aqueous alcohol concentration. In sharp contrast, a continuous variation in their behavior is observed, when their potency is calculated in terms of the amount of alcohol partitioned in the membrane. Our findings strongly support a lipid-mediated mechanism of action for alcohols on rhodopsin and, by analogy, for other G protein-coupled receptors.

Alcohols↗
PubMed1996

A role for phospholipid polyunsaturation in modulating membrane protein function.

Visual transduction is one of the best characterized G protein--coupled signalling systems. In addition, about 50% of the disk membrane phospholipid acyl chains are 22:6n-3, making this system ideal for determining the role of polyunsaturation in modulating membrane-signalling systems. The extent of formation of metarhodopsin II (MII), the G protein--activating photointermediate of rhodopsin, was studied in phospholipid vesicles composed of a variety of phosphatidylcholines, differing in their acyl chain composition at the sn-2 position. The amount of MII formed increased progressively with the level of acyl chain unsaturation at the sn-2 position. The effect of added cholesterol was to reduce the amount of MII formed. The acyl chain packing free volume of the rhodopsin containing lipid vesicles was characterized by a fractional volume parameter fv derived from measurements of the time-resolved fluorescence anisotropy decay of the hydrophobic membrane probe 1,6-diphenyl-1,3,5-hexatriene. The relationship among sn-2 acyl chain unsaturation, cholesterol content, and MII formation is explained on the basis of variation in fv with bilayer lipid composition and a novel model for the packing of phospholipids containing polyenoic acyl chains, such as 22:6n-3.

Fats, Unsaturated↗
PubMed1996

Amphotericin B responsive Scedosporium apiospermum infection in a patient with acute myeloid leukaemia.

A 71 year old man with newly diagnosed acute myelomonocytic leukaemia developed a soft tissue infection of his foot during his first course of chemotherapy. Scedosporium apiospermum was isolated from the lesion, which resolved rapidly on treatment with intravenous amphotericin B despite being resistant in vitro to this agent. This observation suggests that sensitivity testing of S apiospermum should be interpreted with caution and that clinical response may be a better indicator of outcome.

Aged↗
PubMed1996

The dietary intake of children with IDDM.

OBJECTIVE: To assess the dietary intake of children with IDDM and to determine whether the intake meets the current nutritional recommendations for children with IDDM. RESEARCH DESIGN AND METHODS: A total of 66 children with IDDM who were < 10 years of age were recruited from two suburban Pennsylvania hospitals. To collect dietary intake data, subjects were asked, via telephone interview, to complete three random-day 24-h dietary recalls. Data were analyzed for the content of nutrients and other food components by a computerized database program. Intakes were expressed as a 3-day average intake for each subject. RESULTS: Overall mean intake of protein and cholesterol approximated the current recommendations. The mean intake of saturated fat exceeded recommendations, while fiber intake was less than the recommended level. Many of the children consumed levels of saturated fat well above recommendations. Energy, vitamin, and mineral intakes were adequate for the overall sample. However, from 10 to 40% of the sample had an inadequate intake of vitamin D, vitamin E, and zinc. The percentage of those with inadequate intakes of these nutrients decreased with age. CONCLUSIONS: These data suggest that, on average, among this sample of children with IDDM aged < 10 years, adherence to the current nutritional recommendations for children with IDDM was adequate, but some individual children had intakes that were not consistent with the recommendations for optimal management of IDDM.

Child↗
PubMed1994

Effect of ethanol on metarhodopsin II formation is potentiated by phospholipid polyunsaturation.

The role of phospholipids in modulating the effect of ethanol on membrane receptor activation was investigated by studying the extent of metarhodopsin II (MII) formation in vesicles formed POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and PDPC (1-palmitoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine) and in native rod outer segment disk membranes as a function of ethanol concentration. The equilibrium concentration of MII, the G protein-activating form of photoactivated rhodopsin, was found to increase as a function of ethanol concentration in all three bilayers. Phospholipid composition had a marked effect on ethanol potency, with the presence of polyunsaturated phospholipid acyl chains increasing ethanol potency by 40%. The effects of ethanol on lipid acyl chain packing in POPC and PDPC were investigated using frequency domain anisotropy decay measurements of the fluorescent membrane probe 1,6-diphenyl-1,3,5-hexatriene. Enhanced formation of MII due to the presence of ethanol was correlated with the effects of ethanol on acyl chain packing properties. These findings support a phospholipid-mediated mechanism for the action of ethanol in modulating integral membrane receptor conformation.

Cell Membrane↗
PubMed1994

Myelomeningocele at the sacral level. Long-term outcomes in adults.

We reviewed the long-term outcome of thirty-six patients who had a myelomeningocele at the sacral level and whose average age was twenty-nine years (range, nineteen to fifty-one years). The patients were followed at our institution for an average of ten years (range, one to thirty-three years); however, the medical records from birth on were available for all of the patients. Instead of the expected outcome that function had been maintained in this group of patients, we found a decline in the ability to walk of eleven of the thirty-five patients who had been community ambulators initially. At the time of the most recent follow-up examination, five had become household ambulators, two were non-functional ambulators, and four were non-ambulators. The one patient who initially had been a household ambulator was a non-ambulator at the time of the most recent follow-up examination. A decrease in plantar flexion was found in fourteen patients and a decrease in plantar sensation, in fifteen. Breakdown of the skin and soft-tissue infections on the plantar surface of the metatarsal heads and of the heel were seen in twenty-seven and twenty-three patients, respectively, and were related to the absence of plantar sensation. Fifteen patients had osteomyelitis involving the lower extremity. Eleven patients had had a total of fourteen amputations: five involved one toe or more, four involved one ray or more, two were Syme amputations, and three were below-the-knee amputations. By the most recent follow-up examination, thirty-three patients had had a total of 371 orthopaedic procedures. The procedures included tendinous procedures; osteotomies; soft-tissue releases, transfers, and débridements; amputations; and arthrodeses of the lower extremities or spine.

Adult↗
PubMed1993

Low-dose aspirin fails to inhibit increased platelet reactivity in patients with peripheral vascular disease.

A study was performed to evaluate the effect of low-dose aspirin (75 mg/day) on platelet reactivity in patients with peripheral vascular disease. Platelet function was measured in 31 patients with advanced peripheral vascular disease requiring surgery. Shear-induced haemostasis and collagen-induced thrombus formation were used as indicators of platelet reactivity. They were measured in non-anticoagulated fresh whole blood samples ex vivo using a haemostatometer. Results were compared with those from 30 age- and sex-matched controls. Shear-induced haemostasis and collagen-induced thrombus formation were also measured in 18 of the 31 patients before the start of aspirin administration (before operation) and 1 week after surgery. Patients with advanced peripheral vascular disease were found to have increased platelet reactivity with a mean(s.e.m.) shear-induced haemostasis index of 1977(180) mmHg s compared with a control value of 3161(234) mmHg s (P < 0.001) and a mean(s.e.m.) collagen-induced thrombus formation index of 3980(460) mmHg s compared with a control value of 5350(420) mmHg s (P = 0.02). Perioperative low-dose aspirin failed to inhibit platelet function in patients with peripheral vascular disease.

Aged↗
PubMed1993

Beta carotene uptake into atherosclerotic plaque: enhanced staining and preferential ablation with the pulsed dye laser.

The yellow color of atherosclerotic plaque is due to the presence of carotenoids, which absorb light between 430-530 nm and account for the preferential ablation of plaque by the pulsed dye laser operating at 480 nm. This study was designed to examine tissue uptake of beta-carotene and the effect of uptake on arterial plaque ablation. Forty-two atherosclerotic NZW rabbits were given intravenous beta-carotene at a dose of 40 mg/kg, twice weekly and killed between 1 hour and 28 days after the initial injection. beta-carotene was not detected in control specimens but was significantly greater in plaque than in normal wall at all time points following beta-carotene injection (P < 0.04 Mann Whitney U test). The ablation threshold was significantly lower in beta-carotene treated plaque than in untreated plaque or normal arterial wall (P < 0.01, Fisher's exact test). In this model beta-carotene is preferentially taken up into arterial plaque, resulting in increased absorption of laser radiation at 480 nm and enhanced tissue ablation.

Adipose Tissue↗
PubMed1993

The role of fatty acid saturation on plasma lipids, lipoproteins, and apolipoproteins: I. Effects of whole food diets high in cocoa butter, olive oil, soybean oil, dairy butter, and milk chocolate on the plasma lipids of young men.

The present studies were conducted to evaluate the cholesterolemic effects of whole-food diets high in stearic acid. In study no. 1, normocholesterolemic young men were fed diets high in stearic acid provided by cocoa butter (CB); oleic acid provided by olive oil (OO); linoleic acid provided by soybean oil (SO); and myristic acid (and lauric acid) provided by dairy butter (B). In study no. 2, different subjects with similar baseline characteristics were fed diets high in stearic acid provided by milk chocolate (C), CB, CB+B (4:1, MIX), and myristic (and lauric) acid provided by B. Both studies used a randomized, crossover, double-blind experimental design, and experimental subjects (n = 18 for study no. 1 and n = 15 for study no. 2) in each study consumed every diet for 26 days with a 1-month wash-out period between each experimental period. The diets provided 37% of calories from fat, of which 81% was provided by the test fat. Ten ounces (280 g) C was provided daily by the C diet. In study no. 1, the B diet was hypercholesterolemic, whereas the SO diet was hypocholesterolemic, compared with the other diets. The OO and SO diets were hypocholesterolemic compared with the CB diet. Low-density lipoprotein (LDL) cholesterol levels, in general, paralleled the changes in plasma total cholesterol levels. SO significantly decreased apolipoprotein (apo) B levels compared with the other diets. Plasma very-low density lipoprotein (VLDL) cholesterol, high-density lipoprotein (HDL) cholesterol, and apo A-I levels were unaffected by the experimental diets.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1992

Time and cost analysis of a computer-assisted telephone interview system to collect dietary recalls.

This study, conducted in 1991, examined the time requirements and costs of obtaining 24-hour dietary recalls via telephone interviews using the University of Minnesota's microcomputer Nutrient Data System to conduct the interviews and compute the nutrient composition of the diets. The subjects were 156 hypercholesterolemic children (aged 4-10 years) and 102 hypercholesterolemic adults (aged 21-65 years), who were participating in ongoing cholesterol education programs. A total of 391 recalls were completed with the children and 278 with the adults. For each completed interview, 3.5 and 2.8 attempts were required, respectively. Evenings were the most productive time for completing interviews. All tasks associated with completing the interviews (attempts to call, interviews per se, and postinterview procedures) required an average of 39.7 and 35.5 minutes per completed interview with the children and adults, respectively. About half of these total times were actually devoted to conducting the interview. The costs per completed interview were $9.22 for the children versus $6.99 for the adults. This difference reflects the greater number of attempts required to reach the children, the longer duration of their interviews, and the higher intrastate toll rates for calls to them as compared with the interstate rates for calls to the adults.

Adult↗
PubMed1992

Effect of phosphorylation on receptor conformation: the metarhodopsin I in equilibrium with metarhodopsin II equilibrium in multiply phosphorylated rhodopsin.

The superfamily of membrane-bound receptors, which function in signal transduction by activating a guanine nucleotide binding protein or G-protein in response to agonist binding, shares a number of structural and mechanistic properties. Among these similarities is downregulation of functional activity via receptor phosphorylation. In this study, the effects of intermediate levels of phosphorylation (greater than or equal to 4 added phosphates per receptor molecule) on receptor conformational equilibria are examined by comparing the photochemical properties of phosphorylated and unphosphorylated rhodopsins which were incorporated separately into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine vesicles. Postflash spectra reflecting the contributions of metarhodopsins I, II, and III (meta I, meta II, and meta III) were obtained from these samples. Deconvolution of appropriate difference spectra allowed a determination of the concentration of the photointermediates of interest. Meta II is the form of photolyzed rhodopsin which binds and activates the visual G-protein (Gt); thus, its relative abundance at equilibrium and temporal stability are important parameters in determining the efficiency of visual signal transduction. The effects of pH and temperature on the meta I in equilibrium with meta II equilibrium constant (Keq) and the rate of decay of meta II to meta III were examined for the reconstituted phosphorylated and unphosphorylated rhodopsin samples. Keq was essentially unaffected by phosphorylation when measured at pH 7.0 and 8.0 and 20 and 37 degrees C. The decay time (lifetime) of meta II----meta III had a value of approximately 4.7 min in both phosphorylated and unphosphorylated samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Role of sn-1-saturated,sn-2-polyunsaturated phospholipids in control of membrane receptor conformational equilibrium: effects of cholesterol and acyl chain unsaturation on the metarhodopsin I in equilibrium with metarhodopsin II equilibrium.

The effect of phospholipid bilayer acyl chain packing free volume on the equilibrium concentration of the form of photolyzed rhodopsin which initiates visual signal transduction, metarhodopsin II (meta II), is examined in reconstituted systems formed from the saturated phospholipid dimyristoylphosphatidylcholine (DMPC) and in the polyunsaturated phospholipid sn-1-palmitoyl-sn-2-arachidonoylphosphatidylcholine (PAPC) with and without 30 mol% cholesterol. The extent of meta II formation is determined from both flash photolysis measurements and rapidly acquired absorbance spectra. Equilibrium and dynamic properties of the lipid bilayer are characterized by the dynamic fluorescence properties of 1,6-diphenyl-1,3,5-hexatriene (DPH). DPH orientational properties are characterized by fv, a parameter which reflects the volume available for probe reorientation in the bilayer, relative to that available in an unhindered, isotropic environment [Straume, M., & Litman, B. J. (1987) Biochemistry 26, 5121-5126]. The metarhodopsin I in equilibrium with meta II equilibrium constant, Keq has a linear relationship with fv for rhodopsin in PAPC vesicles with and without cholesterol as well as for rhodopsin in DMPC vesicles, and these two correlation lines have different slopes. The correlations between Keq and fv in PAPC and DMPC systems are compared with a similar correlation in the native rod outer segment disk membrane and one reported previously in an egg phosphatidylcholine (egg PC) system [Mitchell, D. C., Straume, M., Miller, J. L., & Litman, B. J. (1990) Biochemistry 29, 9143-9149].(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Lasers in surgery.

In January 1991 the winter meeting of the Surgical Research Society was held at St. Bartholomew's Hospital and the Institute of Education, London. During the meeting a symposium was held entitled 'Shedding light on lasers'. Speakers addressed the general principles of lasers, laser-tissue interactions and the applications of lasers in gastro-enterology and in vascular disease. This was followed by an open discussion of the current indications, complications and outcome, together with future possible applications for lasers in medicine.

Arterial Occlusive Diseases↗
PubMed1992

Laser-assisted angioplasty for arterial occlusion of the lower limb: initial results and follow-up.

A flashlamp-pumped pulsed dye laser operating at either 480 or 504 nm, coupled to an integral ball-tipped optical fibre, was used to recanalize occluded lower limb arteries. All channels created by the laser were augmented with balloon dilatation. We have treated 78 limbs in 71 patients; 46 limbs (59 per cent) had rest pain and 22 (48 per cent) of these had tissue loss. The median occlusion length was 18 (range 0.5-58) cm. Technical success was achieved in 58 limbs (74 per cent) with clinical success in 46 (59 per cent). Success rates fell with increasing length of occlusion. Two patients died in the perioperative period. A subgroup of 22 patients with marked discrete arterial calcification had a lower technical success rate than the subgroup without calcification (50 per cent versus 84 per cent, P less than 0.01). Both subgroups displayed a similar pattern of reclosure during follow-up. The cumulative patency rate after technical success was 67 per cent at 6 months and 45 per cent at 12 months. Forty-six (59 per cent) limbs avoided bypass surgery or amputation. Laser-assisted angioplasty may offer an alternative to femoropopliteal bypass, although the former procedure is not as durable.

Adult↗
PubMed1991

Functional equivalence of metarhodopsin II and the Gt-activating form of photolyzed bovine rhodopsin.

Absorption of a photon by the visual pigment rhodopsin leads to the formation of an activated conformational state, denoted rho*, which is capable of activating the visual G-protein, Gt. The bleaching of rhodopsin can be resolved into a series of spectrally distinct photointermediates. Previous studies suggest that the photointermediate metarhodopsin II (meta II, lambda max of 380 nm) corresponds to the physiologically active form rho*. In the studies reported herein, spectral and enzymological data were analyzed and compared so as to evaluate the temporal correspondence between meta II and rho*. This information was obtained by direct observation of the meta II and rho* decay times in parallel experiments utilizing identical preparations of urea-stripped, bovine retinal rod outer segment disk membranes at pH 8.0, 20 degrees C. Postflash spectra were deconvolved to resolve the meta II absorbance at 380 nm, and a decay time for the loss of meta II of 8.2 min (SD = 0.5 min) was obtained from fitting these data to a single-exponential decay process. The diminishing ability of bleached rhodopsin to activate Gt was measured by monitoring the level of catalyzed exchange of Gt-bound GDP for a nonhydrolyzable GTP analogue. Analysis of the decrease in the initial velocity of nucleotide exchange, measured at various postflash incubation times, yielded a rho* decay time of 7.7 min (SD = 0.5 min) when analyzed as a single-exponential process. The similarity of these decay times provides direct evidence that meta II and rho* are present over the same time regime, and further supports the equivalence of these two forms of photoactivated rhodopsin.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗