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Biomedical subjects

S A Ross

Publications and source records attributed to S A Ross.

At least 55 records · Page 3Linked to original sources
PubMed1999

Cloning and preliminary characterization of a 121 kDa protein with multiple predicted C2 domains.

In the course of characterizing proteins present in a preparation of vesicles from rat adipocytes containing glucose transporters, we examined a protein that migrated at 115 kDa upon SDS gel electrophoresis (designated vp115). Sequences of tryptic peptides were obtained, and from this information the cDNA for rat vp115 was cloned. The cDNA encodes an open reading frame for a protein of 121 kDa. Computer-aided sequence analysis predicted that vp115 has a potential membrane-inserted or membrane-spanning domain toward its amino terminus, followed by five C2 domains. Immunoblotting revealed that vp115 was not actually a component of the glucose transporter-containing vesicles, was most abundant in the plasma membranes and high density microsome fractions of rat adipocytes, and was expressed in all the major rat tissues.

Adipocytes↗
PubMed1999

Cloning and characterization of a 22 kDa protein from rat adipocytes: a new member of the reticulon family.

In the course of our examination of proteins associated with the GLUT4-containing vesicles of rat adipocytes we have identified a new 22 kDa member of the family of endoplasmic reticulum (ER) proteins known as reticulons. The protein, which we refer to as vp20, was purified from a preparation of GLUT4-containing vesicles of rat adipocytes, and tryptic peptides were micro-sequenced. From this information a cDNA encoding a single open reading frame for a protein of 22 kDa was cloned. This protein is homologous to known members of the reticulon protein family. vp20 has two hydrophobic stretches of about 35 amino acids that could be membrane spanning domains and an ER retention motif at its carboxy-terminus. vp20 was most abundant in the high density microsome fraction of adipocytes, which is the fraction most enriched in ER. Only a small fraction of vp20 was present in the GLUT4 vesicle population, and that fraction appears to be due to ER vesicles that were non-specifically bound to the adsorbent. Analysis of tissue distribution of vp20 in rats revealed that it is concentrated in muscle, fat and the brain.

Adipocytes↗
PubMed1999

Flavonoids from Maclura tinctoria.

Seven flavonoids, including two new natural products, were isolated from an ethanol extract of the bark of Maclura tinctoria (L.) Gaud. The new compounds are steppogenin 4'-O-beta-D-glucoside and orobol 5,3'-di-O-methyl-8-C-glucoside. Orobol, steppogenin, aromadendrin, dihydromorin and orobol 7-O-beta-D-glucoside were also isolated.

Flavonoids↗
PubMed1999

Day-to-day consistency in amount and source of carbohydrate intake associated with improved blood glucose control in type 1 diabetes.

OBJECTIVE: To determine if a relationship exists between blood glucose control and variability in nutrient intake from day-to-day in subjects with type 1 diabetes. METHODS: Two three-day diet records and one measurement of glycated hemoglobin (HbA1c) were obtained from 272 subjects with type 1 diabetes treated with a mixture of regular and NPH insulins before breakfast and supper and using a standardized algorithm to adjust insulin dose according to the results of self-monitoring of blood glucose two to four times daily. Day-to-day variation in nutrient intake was expressed as the coefficient of variation (CV = SDx100/mean). RESULTS: Nutrient intakes in the study population (mean +/- SD) were energy 8.35+/-2.43 MJ, fat 81+/-30 g, protein 94+/-28 g, carbohydrate 227+/-68 g, starch 126+/-38 g and dietary fiber 20+/-6 g with diet glycemic index being 84.2+/-7.4. Neither energy, nutrient intakes nor insulin dose was significantly related to HbA1c. Day-to-day variation of carbohydrate (p = 0.0097) and starch (p = 0.0016) intakes and diet glycemic index (p = 0.033) was positively related to HbA1c, and the associations remained significant when adjusted for age, sex, duration of diabetes and BMI. Day-to-day variation in energy, protein or fat intakes was not related to HbA1c. CONCLUSIONS: Consistency in the amount and source of carbohydrate intake from day-to-day is associated with improved blood glucose control in people with type 1 diabetes, a result which supports continued educational efforts to achieve adherence to a diabetes diet plan. This conclusion may not apply to people on intensified insulin therapy who adjust their insulin dose based on their actual carbohydrate intake at each meal.

Adolescent↗
PubMed1999

Changes in ankle spasticity and strength following selective dorsal rhizotomy and physical therapy for spastic cerebral palsy.

OBJECT: In this investigation the authors quantified changes in ankle plantarflexor spasticity and strength following selective dorsal rhizotomy (SDR) and intensive physical therapy in patients with cerebral palsy (CP). METHODS: Twenty-five patients with cerebral palsy (CP group) and 12 able-bodied volunteers (AB controls) were tested with a dynamometer. For the spasticity measure, the dynamometer was used to measure the resistive torque of the plantarflexors during passive ankle dorsiflexion at five different speeds. Data were processed to yield a single value that simultaneously encompassed the three key elements associated with spasticity: velocity, resistance, and stretch. For the strength test, the dynamometer rotated the ankle from full dorsiflexion to full plantarflexion while a maximum concentric contraction of the plantarflexors was performed. Torque angle data were processed to include the work done by the patient or volunteer on the machine. Plantarflexor spasticity values for the CP group were significantly greater than similar values for the AB control group prior to surgery but not significantly different after surgery. Plantarflexor strength values of the CP group were significantly less than those of the AB control group pre- and postsurgery. Postsurgery strength values did not change relative to presurgery values. CONCLUSIONS: The spasticity results of the present investigation agreed with those of previous studies indicating a reduction in spasticity for the CP group. The strength results did not agree with the findings of most previous related literature, which indicated that a decrease in strength should have occurred. The strength results agreed with a previous investigation in which knee flexor strength was objectively examined, indicating that strength did not decrease as a consequence of an SDR. The methods of this investigation could be used to improve SDR patient selection.

Adolescent↗
PubMed1998

Association of insulin receptor substrate 3 with SH2 domain-containing proteins in rat adipocytes.

We have recently purified and cloned a new member of the insulin receptor substrate family, designated insulin receptor substrate 3 (IRS-3), from rat adipocytes. The amino acid sequence of IRS-3 shows multiple potential sites for tyrosine phosphorylation in motifs which engage specific SH2 domain-containing proteins. In order to determine which SH2 domain proteins complex with IRS-3, we have searched for coimmunoprecipitation from lysates of untreated and insulin-stimulated adipocytes. Phosphatidylinositol 3-kinase and the tyrosine phosphatase SHP-2 complexed with the tyrosine phosphorylated form of IRS-3, whereas the phospholipase Cgamma did not, and the adaptor Grb2 did so to a much lesser extent. These findings complete the survey of SH2 domain proteins associated with each of the four known members of the IRS family and provide the framework for further analysis of the role of IRS-3 in insulin signaling.

Adaptor Proteins, Signal Transducing↗
PubMed1998

Increased intracellular sequestration of the insulin-regulated aminopeptidase upon differentiation of 3T3-L1 cells.

In fat and muscle cells, the glucose transporter GLUT4 is sequestered in an intracellular compartment under basal conditions and redistributes markedly to the plasma membrane in response to insulin. Recently, we characterized a membrane aminopeptidase, designated IRAP (insulin-regulated aminopeptidase), that colocalizes with intracellular GLUT4 and similarly redistributes markedly to the plasma membrane in response to insulin in adipocytes. In contrast to GLUT4, IRAP is also expressed in 3T3-L1 fibroblasts, and this finding provided an opportunity to compare its subcellular distribution in fibroblasts and adipocytes. The relative amount of IRAP at the cell surface was measured by a cell surface biotinylation method. The portion of total IRAP at the cell surface in unstimulated adipocytes was 30% of that in unstimulated fibroblasts. Upon insulin treatment the portion of IRAP at the cell surface was the same in fibroblasts and adipocytes, and was increased 1.8-fold in fibroblasts and 8-fold in adipocytes. A similar analysis of the distribution of the transferrin receptor (TfR), the paradigm for recycling plasma membrane receptors, revealed that the portions of the TfR at the cell surface in both the basal and insulin-treated states were almost unchanged upon differentiation, and that insulin caused an increase of about 1. 6-fold in the amount of TfR at the cell surface. These results show that enhanced intracellular sequestration of IRAP occurs during adipogenesis, and that this effect underlies the larger insulin-elicited fold increase of IRAP at the cell surface in adipocytes.

3T3 Cells↗
PubMed1998

Sortilin is the major 110-kDa protein in GLUT4 vesicles from adipocytes.

Vesicles containing the glucose transporter GLUT4 from rat adipocytes contain a major protein of 110 kDa. We have isolated this protein, obtained the sequences of peptides, and cloned a large portion of its cDNA. This revealed that the protein is sortilin, a novel membrane protein that was cloned in another context from a human source while this work was in progress. Subcellular fractionation of rat and 3T3-L1 adipocytes, together with GLUT4 vesicle isolation, showed that sortilin was primarily located in the low density microsomes in vesicles containing GLUT4. Insulin caused a 1.7-fold increase in the amount of sortilin at the plasma membranes of 3T3-L1 adipocytes, as assessed by cell surface biotinylation. The expression of sortilin in 3T3-L1 cells occurred only upon differentiation. Previous characterization of sortilin has led to the suggestion that it functions to sort lumenal proteins from the trans Golgi. The significance of its insulin-stimulated increase at the cell surface and of its expression upon differentiation will require definitive delineation of its function.

3T3 Cells↗
PubMed1998

Spasticity and strength changes as a function of selective dorsal rhizotomy.

This investigation quantified pre- and postsurgery (8 months) hamstring muscle spasticity and strength in children with cerebral palsy (CP) undergoing a selective dorsal rhizotomy. Nineteen children with CP (CP group) and six children with able bodies (AB group) underwent testing with a dynamometer. For the spasticity measure, the dynamometer measured the resistive torque of the hamstring muscles during passive knee extension at four different speeds. Torque angle data were processed to calculate the work done by the machine to extend the knee for each speed. Linear regression was used to calculate the slope of the line of best fit for the work velocity data. The slope simultaneously encompassed three key elements associated with spasticity (velocity, resistance, and stretch) and was considered the measure of spasticity. For the strength test, the dynamometer moved the leg from full knee extension to flexion while a maximum concentric contraction of the hamstring muscles was performed. Torque angle data were processed to calculate the work done on the machine by the child. Hamstring spasticity values for the CP group were significantly greater than similar values for AB group prior to surgery; however they were not significantly different after surgery. Hamstring strength values for the CP group remained significantly less than those for the AB group after surgery, but were significantly increased relative to their presurgery values. The results of spasticity testing in the present investigation agreed with previous studies indicating a reduction in spasticity for the CP group. The results of strength testing did not agree with those in the previous literature; a significant increase in strength was observed for the CP group.

Journal Article↗
PubMed1998

Early and late post-ischaemic recovery of contractile function is affected to different degrees by isoflurane and halothane in the anaesthetized rabbit model.

The protective efficacy of halogenated anaesthetics on myocardial injury has never been compared during early reperfusion and late reperfusion in an in vivo animal model. We compared recovery of left ventricular function under isoflurane (0.5 MAC) and halothane (0.5 MAC) anaesthesia after a brief period of regional ischaemia (15 min) in acutely instrumented rabbits. Rabbits were instrumented for the measurement of regional segment length and left ventricular pressure. Rabbits receiving isoflurane showed greater recovery of systolic shortening fraction (%SS) both during early and late reperfusion compared with halothane anaesthesia. Isoflurane protected the post-ischaemic myocardium to a greater extent than halothane anaesthesia. Early recovery of contractile function may be a predictor of contractile recovery during the later stages of reperfusion.

Anesthetics, Inhalation↗
PubMed1998

Quality of reviews in epidemiology.

OBJECTIVES: This study examined the quality of recent reviews in epidemiology. METHODS: All 1995 issues of 7 widely read epidemiology journals were searched to identify reviews. RESULTS: Twenty-nine reviews were identified. Methodology was not specified or incomplete for literature searches in 79% of reviews; the same was true for inclusion criteria in 83% and for combining studies in 62%. More than 60% of the reviews were not methodologically systematic. CONCLUSIONS: There is a need to improve the quality of review papers in epidemiology. If systematic methodology were followed more frequently, epidemiologic science and its application could be improved.

Epidemiology↗
PubMed1998

No relationship between carbohydrate intake and effect of acarbose on HbA1c or gastrointestinal symptoms in type 2 diabetic subjects consuming 30-60% of energy from carbohydrate.

OBJECTIVE: To determine the relationship between carbohydrate intake and the effect of acarbose on HbA1c in subjects with type 2 diabetes treated with acarbose alone, acarbose plus sulfonylurea, acarbose plus metformin, or acarbose plus insulin. RESEARCH DESIGN AND METHODS: We conducted a double-blind randomized placebo-controlled study in which subjects with diabetes in four treatment strata (77 on diet alone, 83 treated with metformin, 103 treated with sulfonylurea, and 91 treated with insulin) were randomized to treatment with placebo or acarbose for 12 months. Before randomization, and 3, 6, 9, and 12 months after randomization, fasting blood was obtained for HbA1c, and 3-day diet records were collected. Subjects who completed at least 6 months of acarbose therapy and provided at least three 3-day diet records were included. RESULTS: In the 114 subjects included in this analysis, carbohydrate intake varied from approximately 30-60% of energy There was no significant relationship between carbohydrate intake and change in HbA1c in any of the four treatment strata (diet: n=26, r=0.35, P=0.076; metformin: n=27, r=0.26, P=0.19; sulfonylurea: n=35, r=0.24, P=0.16; insulin: n=25, r=-0.27, P=0.19). In the 80 subjects consuming <50% of energy from carbohydrate, the fall in HbA1c (7.83 +/-0.17% at baseline to 6.72+/-0.13% on acarbose, P < 0.001) was no different from that of the 34 subjects consuming >50% of energy from carbohydrate (7.55+/-0.25% at baseline to 6.66+/-0.23% on acarbose, P < 0.001). There was no difference in carbohydrate intake between those who dropped out of the study because of gastrointestinal side effects and those who did not, and there was no relationship between severity of symptoms and the composition of the diet. CONCLUSIONS: In subjects with type 2 diabetes consuming 30-60% of energy from carbohydrate, the effect of acarbose on HbA1c and gastrointestinal symptoms was not related to carbohydrate intake. Because most people consume at least 30% of energy from carbohydrate, we conclude that no special diet is needed for acarbose to be effective in improving blood glucose control in the treatment of type 2 diabetes.

Acarbose↗
PubMed1998

Variation of postprandial plasma glucose, palatability, and symptoms associated with a standardized mixed test meal versus 75 g oral glucose.

OBJECTIVE: To compare within-subject variability of plasma glucose measured 2 h after a glucose tolerance test (GTT) with that of plasma glucose measured 2 h after administration of a standardized test meal (diabetes screening product [DSP], Ceapro, Edmonton, Alberta, Canada) and to determine the relationship between the two sets of plasma glucose measurements. RESEARCH DESIGN AND METHODS: Plasma glucose and insulin responses of 36 overnight-fasted subjects (10 lean normal, 9 obese normal, 9 with impaired glucose tolerance [IGT], and 8 with mild diabetes) were studied on eight different mornings after they consumed 75 g oral glucose or 50 g carbohydrate from the DSP. Each test meal was repeated four times by each subject. Within-subject coefficients of variation (CVs) (CV = 100 x SD/mean) of plasma glucose concentrations 2 h after administration of the GTT and DSP were compared by repeated measures ANOVA and linear regression analysis. RESULTS: Mean plasma glucose 2 h after administration of the DSP (D) was linearly related to that 2 h after the GTT (G): G = 1.5 x D - 1.6 (r = 0.97, P < 0.0001). The CV of 2-h plasma glucose was significantly lower after administration of the DSP, 10.5 +/- 1.0%, than after the GTT, 12.7 +/- 1.18% (P = 0.025). The effect of test meal on CV differed in different groups of subjects (P = 0.018), with the largest difference found in IGT subjects, in whom the CV after DSP administration was 47% less than after the GTT (P = 0.0005). The DSP was significantly more palatable and produced fewer adverse symptoms than the GTT. CONCLUSIONS: Plasma glucose concentrations measured 2 h after DSP administration are closely related to those measured 2 h after the GTT but are more consistent than the 2-h post-GTT concentrations within the critical IGT range. This finding suggests that measurement of plasma glucose 2 h after administration of the DSP may allow more precise discrimination among normal glucose levels, IGT, and diabetes than measurement of plasma glucose 2 h after the GTT.

Administration, Oral↗
PubMed1998

Spasticity and strength changes as a function of selective dorsal rhizotomy.

OBJECT: The goal of this investigation was to quantify changes in hamstring muscle spasticity and strength in children with cerebral palsy (CP) as a function of their having undergone a selective dorsal rhizotomy. METHODS: Nineteen children with CP (CP group) and six children with able bodies (AB group) underwent testing with a dynamometer. For the spasticity measure, the dynamometer measured the resistive torque of the hamstring muscles during passive knee extension at four different speeds. Torque-angle data were processed to calculate the work done by the machine to extend the knee for each speed. Linear regression was used to calculate the slope of the line of best fit for the work-velocity data. The slope simultaneously encompassed three key elements associated with spasticity (velocity, resistance, and stretch) and was considered the measure of spasticity. For the strength test, the dynamometer moved the leg from full knee extension to flexion while a maximum concentric contraction of the hamstring muscles was performed. Torque-angle data were processed to calculate the work done on the machine by the child. Hamstring spasticity values for the CP group were significantly greater than similar values for the AB group prior to surgery; however, they were not significantly different after surgery. Hamstring strength values for the CP group remained significantly less than those for the AB group after surgery, but were significantly increased relative to their presurgery values. CONCLUSIONS: The results of spasticity testing in the present investigation agreed with those of previous studies, indicating a reduction in spasticity for the CP group. The results of strength testing did not agree with those in the previous literature; a significant increase in strength was observed for the CP group.

Adolescent↗
PubMed1997

Trafficking kinetics of the insulin-regulated membrane aminopeptidase in 3T3-L1 adipocytes.

In fat and muscle cells insulin causes the marked translocation of the glucose transporter GLUT4 from its intracellular location to the plasma membrane. We and others have discovered an insulin-regulated membrane aminopeptidase (designated IRAP) that colocalizes with intracellular GLUT4 and also translocates markedly in response to insulin. This study describes the trafficking kinetics of IRAP in 3T3-L1 adipocytes. By means of a surface biotinylation method, the half-time for the increase in IRAP at the plasma membrane in response to insulin was found to be 2 min. The increase was completely blocked by the phosphatidylinositol 3-kinase inhibitor, wortmannin. In insulin-treated cells, biotinylated IRAP, initially at the plasma membrane, equilibrated with the intracellular pool with a half-time of 2 min. Thus, IRAP continuously recycles. Finally, vesicles isolated from the intracellular membranes with antibodies against IRAP and GLUT4 showed the same protein composition. In conjunction with results in the literature, these findings indicate that IRAP and GLUT4 traffic through the same intracellular compartments.

3T3 Cells↗
PubMed1997

Membrane amine oxidase cloning and identification as a major protein in the adipocyte plasma membrane.

A 97-kDa protein present in the glucose transporter (GLUT4 isotype)-containing vesicles from rat adipocytes has been isolated, the sequences of two tryptic peptides were obtained, and on the basis of these its cDNA partially cloned. The 97-kDa protein is almost certainly identical to a major integral glycoprotein of this size in the rat adipocyte plasma membrane, since its predicted N-terminal sequence is the same as that recently determined for this glycoprotein by amino acid sequencing. Moreover, the predicted partial sequence (322 amino acids) of the 97-kDa protein is highly homologous to the corresponding region of a human placental amine oxidase, which was cloned simultaneously and proposed to be a secreted protein. The amino acid sequence of the 97-kDa rat/human amine oxidase indicates that the protein consists of a very short N-terminal cytoplasmic domain followed by a single transmembrane segment and a large extracellular domain containing the catalytic site. Thus this study establishes the 97-kDa rat/human amine oxidase as the first integral membrane amine oxidase to be cloned. The membrane amine oxidase was more abundant in the plasma membranes than the low density microsomes of the adipocyte, and in contrast to some other proteins found in GLUT4 vesicles, it did not redistribute to the plasma membrane in response to treatment of the cells with insulin.

Adipocytes↗