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Biomedical subjects

F Tremblay

Publications and source records attributed to F Tremblay.

At least 19 recordsLinked to original sources

Amino acid and insulin signaling via the mTOR/p70 S6 kinase pathway. A negative feedback mechanism leading to insulin resistance in skeletal muscle cells.

Amino acids have emerged as potent modulators of the mTOR/p70 S6 kinase pathway. The involvement of this pathway in the regulation of insulin-stimulated glucose transport was investigated in the present study. Acute exposure (1 h) to a balanced mixture of amino acids reduced insulin-stimulated glucose transport by as much as 55% in L6 muscle cells. The effect of amino acids was fully prevented by the specific mTOR inhibitor rapamycin. Time course analysis of insulin receptor substrate 1 (IRS-1)-associated phosphatidylinositol (PI) 3-kinase activity revealed that incubation with amino acids speeds up its time-dependent deactivation, leading to a dramatic suppression (-70%) of its activity after 30 min of insulin stimulation as compared with its maximal activation (5 min of stimulation). This accelerated deactivation of PI 3-kinase activity in amino acid-treated cells was associated with a concomitant and sustained increase in the phosphorylation of p70 S6 kinase. In marked contrast, inhibition of mTOR by rapamycin maintained PI 3-kinase maximally activated for up to 30 min. The marked inhibition of insulin-mediated PI 3-kinase activity by amino acids was linked to a rapamycin-sensitive increase in serine/threonine phosphorylation of IRS-1 and a decreased binding of the p85 subunit of PI 3-kinase to IRS-1. Furthermore, amino acids were required for the degradation of IRS-1 during long term insulin treatment. These results identify the mTOR/p70 S6 kinase signaling pathway as a novel modulator of insulin-stimulated glucose transport in skeletal muscle cells.

Amino Acids↗

Development and application of a biological peptide pump for the study of the in vivo actions of angiotensin peptides.

In the past few years, a great deal of interest has been focused on the possibility that angiotensin peptides could have direct effects on target tissues independent of their hemodynamic effects. In addition, there has been much speculation on the potential biological roles of angiotensin peptides other than angiotensin II. Unfortunately, a direct test of these possibilities in whole animals has been difficult due to limitations in existing biological systems. In this review, we describe the characteristics of an engineered protein capable of directing the production of a wide variety of peptides to specific tissues and cell types in transgenic animals and discuss its potential applications.

Angiotensins↗

Modulation of corticospinal excitability during imagined knee movements.

In this study, we investigated corticospinal excitability during mental simulation of a leg extension movement with the technique of transcranial magnetic stimulation. Motor evoked potentials were recorded in both knee extensors (quadriceps) and flexors (biceps femoris) in 19 trained participants (healthy volunteers). The amplitude and latency of motor evoked potentials were compared in three conditions: (1) at rest, (2) during motor imagery, and (3) at rest, immediately after motor imagery. The results showed a significant effect (p < 0.001) of conditions on motor evoked potentials amplitude in the quadriceps but not in the biceps femoris. During motor imagery, the size of motor evoked potentials in the quadriceps increased significantly (p < 0.001) compared with rest and post-imagery conditions. Changes in motor evoked potentials latency across conditions were not significant, however. These results are consistent with previous studies in the upper limb and suggest that corticospinal excitability can be enhanced during motor imagery to facilitate responses in specific lower limb muscles.

Adolescent↗

Prevention of skeletal muscle insulin resistance by dietary cod protein in high fat-fed rats.

In the present study, we tested the hypothesis that fish protein may represent a key constituent of fish with glucoregulatory activity. Three groups of rats were fed a high-fat diet in which the protein source was casein, fish (cod) protein, or soy protein; these groups were compared with a group of chow-fed controls. High-fat feeding led to severe whole body and skeletal muscle insulin resistance in casein- or soy protein-fed rats, as assessed by the euglycemic clamp technique coupled with measurements of 2-deoxy-D-[(3)H]glucose uptake rates by individual tissues. However, feeding cod protein fully prevented the development of insulin resistance in high fat-fed rats. These animals exhibited higher rates of insulin-mediated muscle glucose disposal that were comparable to those of chow-fed rats. The beneficial effects of cod protein occurred without any reductions in body weight gain, adipose tissue accretion, or expression of tumor necrosis factor-alpha in fat and muscle. Moreover, L6 myocytes exposed to cod protein-derived amino acids showed greater rates of insulin-stimulated glucose uptake compared with cells incubated with casein- or soy protein-derived amino acids. These data demonstrate that feeding cod protein prevents obesity-induced muscle insulin resistance in high fat-fed obese rats at least in part through a direct action of amino acids on insulin-stimulated glucose uptake in skeletal muscle cells.

Adipose Tissue↗

Defective insulin-induced GLUT4 translocation in skeletal muscle of high fat-fed rats is associated with alterations in both Akt/protein kinase B and atypical protein kinase C (zeta/lambda) activities.

The cellular mechanism by which high-fat feeding induces skeletal muscle insulin resistance was investigated in the present study. Insulin-stimulated glucose transport was impaired ( approximately 40-60%) in muscles of high fat-fed rats. Muscle GLUT4 expression was significantly lower in these animals ( approximately 40%, P < 0.05) but only in type IIa-enriched muscle. Insulin stimulated the translocation of GLUT4 to both the plasma membrane and the transverse (T)-tubules in chow-fed rats. In marked contrast, GLUT4 translocation was completely abrogated in the muscle of insulin-stimulated high fat-fed rats. High-fat feeding markedly decreased insulin receptor substrate (IRS)-1-associated phosphatidylinositol (PI) 3-kinase activity but not insulin-induced tyrosine phosphorylation of the insulin receptor and IRS proteins in muscle. Impairment of PI 3-kinase function was associated with defective Akt/protein kinase B kinase activity (-40%, P < 0.01) in insulin-stimulated muscle of high fat-fed rats, despite unaltered phosphorylation (Ser473/Thr308) of the enzyme. Interestingly, basal activity of atypical protein kinase C (aPKC) was elevated in muscle of high fat-fed rats compared with chow-fed controls. Whereas insulin induced a twofold increase in aPKC kinase activity in the muscle of chow-fed rats, the hormone failed to further increase the kinase activity in high fat-fed rat muscle. In conclusion, it was found that GLUT4 translocation to both the plasma membrane and the T-tubules is impaired in the muscle of high fat-fed rats. We identified PI 3-kinase as the first step of the insulin signaling pathway to be impaired by high-fat feeding, and this was associated with alterations in both Akt and aPKC kinase activities.

Animals↗

New short peptide substrates of endothelin-converting enzyme and characterization of the enzyme.

Endothelin (ET) is a 21 amino acid peptide produced following the cleavage of its precursor, big ET, by a metalloprotease, the endothelin-converting enzyme (ECE). In the study reported here we determined the minimal peptide sequence of big ET necessary for enzyme recognition and cleavage at the P1-P1' site. Furthermore, we have explored the role of the amino acids found at the boundaries of the cleavage site. To reach these goals. we synthesized a series of fragments, all containing the P1-P1' cleavage site, Trp21-Val22. Following the incubation of peptide fragments with a partly purified bovine ECE preparation and after analyzing the cleavage pattern by high-performance liquid chromatography (HPLC), we were able to identify big ET(18-23) amide as the minimal peptide core recognized and cleaved by the enzyme. This hydrolysis was inhibited by phosphoramidon but not by thiorphan, a characteristic of the ECE metalloprotease. However, none of the shorter peptides was able to inhibit the cleavage of big ET-1 by ECE, suggesting that they are not recognized by the enzyme. Particularly, it appears that aspartic acid 18 is a key residue for the recognition phenomenon. The delineation of the minimal structure will be a useful tool to further characterize ECE.

Animals↗

The use of binocular visual acuity in the assessment of intermittent exotropia.

BACKGROUND: It has been suggested that a decrease in distance stereoacuity in patients with intermittent exotropia is a good indicator of diminishing control. However, there has been no adequate explanation for this reported reduction in distance stereoacuity in these patients. We postulate that the decrease in stereoacuity is related to blurred visual acuity created by an increasing demand on accommodation, which these patients use in an attempt to control the exodeviation. This can best be assessed by measuring binocular visual acuity (BVA). Analysis of BVA could provide a useful clinical tool to evaluate control measures used by patients with intermittent exotropia. METHODS: A prospective study of patients with intermittent exotropia, ranging in age from 6 to 60 years, was performed. Only those patients with the presence of either basic or divergence excess (simulated or true) type exodeviation were included in the study. The data analysis included the age of these patients, age at onset of the deviation, monocular and binocular visual acuity, oculomotor and fusional status, and near and distance stereoacuity. RESULTS: Data from 36 patients show that the measurements of BVA correlated well with a corresponding loss of distance stereoacuity but not with the size of the deviation. CONCLUSION: The decrease of stereoacuity reported in patients with exotropia can be explained by increased accommodation and decreased distance BVA. This measurement can be a simple method of quantifying the fusional control of patients with intermittent exotropia.

Accommodation, Ocular↗

Effects of PAX6 mutations on retinal function: an electroretinographic study.

PURPOSE: To investigate the retinal function in aniridic patients with documented PAX6 mutations to determine the range of electroretinogram abnormalities in aniridic patients and to relate electroretinogram findings with specific PAX6 mutations. METHODS: Eleven patients with typical aniridia and fully characterized PAX6 mutations underwent electroretinography. RESULTS: In all 11 patients, electroretinogram recordings were abnormal, ranging from mild to severe. Rod-related and cone-related activities were equally affected. The amplitude of the oscillatory potentials was the most reduced, followed by the b-wave, then to a milder degree the a-wave. Mutations affecting the paired domain of the PAX6 protein had the biggest impact on the electroretinogram amplitudes. Implicit times were increased in a subgroup with mutations affecting only the homeodomain. CONCLUSION: Patients with aniridia have varying degree of retinal dysfunction, ranging from severely abnormal to almost normal. The paired domain appears to have more impact on retinal function than other regions of the PAX6 protein. It is unclear whether mutations affecting the homeodomain lead to alteration of the photoreceptor function.

Adolescent↗

Genotype/phenotype correlations in aniridia.

PURPOSE: To detect and characterize mutations in cases of familial and sporadic aniridia in Maritime Canada, and to look for indications of genotype/phenotype correlation within the cohort. METHODS: Twelve consecutive and unrelated patients (probands) who had total or nearly complete absence of irides, and four affected relatives, were recruited from Maritime Canada. Clinical data were obtained by chart review and electroretinogram testing. Mutations in the PAX6 gene were detected by single-strand conformation polymorphism and characterized by sequence analysis. RESULTS: Eleven different PAX6 mutations, 10 of which are novel, were found. The four patients with congenital cataracts all had mutations in the C-terminal proline-serine-threonine (PST)-rich domain of the PAX6 protein. Electroretinograms of nine of 11 patients displayed depressed scotopic maximum response b-wave amplitudes. The greatest decrease in b-wave amplitudes was seen in patients in whom the paired domain was disrupted by mutation. CONCLUSION: Some aspects of the phenotype of aniridia appear to correlate with the predicted effect of point mutations on the paired and PST domains of the PAX6 protein.

Adolescent↗

Neuronal encoding of texture changes in the primary and the secondary somatosensory cortical areas of monkeys during passive texture discrimination.

Two rhesus monkeys were trained to discriminate, with the use of passive touch, a standard surface [rectangular arrays of raised dots with a spatial period (SP) of 2 mm across the rows and columns] from three modified surfaces in which the SP between rows was increased to 3, 4, or 5 mm over the second half of the surface. After the surface presentation (to digit tips 3 and 4 of one hand) the monkeys indicated the presence or absence of a change in texture by pulling or pushing a lever, respectively, with the opposite hand. Of 193 neurons recorded from primary somatosensory cortex (SI, 3 hemispheres) and 94 neurons from secondary somatosensory cortex (SII, 1 hemisphere), all contralateral to the stimulated hand, the discharge of 51 SI and 19 SII neurons was classified as texture related. Two types of texture-related responses were obtained. Graded neurons showed a linear relationship between mean discharge frequency and SP; nongraded neurons showed a significant change in discharge over the modified half of the surfaces but the discharge did not distinguish between the three modified surfaces. The distribution of these texture responses was significantly different in SI and SII: whereas most of the texture-related neurons in SI (44 of 51, 86%) were graded, the majority of those in SII (12 of 19, 63%) were nongraded. The results were interpreted as suggesting that the nongraded responses reflect feature extraction in SII, signaling the presence of a change in texture but not its magnitude, and so support the notion that texture signals are processed sequentially, first in SI and then in SII.

Animals↗

Cortical mechanisms underlying tactile discrimination in the monkey. I. Role of primary somatosensory cortex in passive texture discrimination.

1. The discharge patterns of 359 single neurons in the hand representation of primary somatosensory cortex (SI) of two monkeys (Macaca mulatta) were recorded during the performance of a passive texture discrimination task with the contralateral hand (104 in area 3b, 149 in area 1, and 106 in area 2). Three nyloprint surfaces were mounted on a drum that was rotated under the digit tips. One surface was entirely smooth, whereas the other two were smooth over the first half and rough over the second half (smooth/ rough) (raised dots, 1 mm high and 1 mm diam, in a rectangular array; spatial period of 3 mm across the rows and columns for most recordings; 9 mm between columns for selected recordings). The monkeys were trained to distinguish between the smooth and smooth/rough surfaces. After the surface presentation, the monkey indicated the texture of the second half of the surface by pushing or pulling, respectively, on a lever with the other arm. For most recordings an average tangential speed of 49 mm/s was tested. For selected recordings motor speed was incremented (63, 75, or 89 mm/s). 2. Two hundred eighty-three neurons had a cutaneous receptive field (RF) on the hand (96 in area 3b, 120 in area 1, and 67 in area 2). Thirty-five neurons had a deep RF (4 in area 3b, 15 in area 1, and 16 in area 2). Seven neurons had mixed cutaneous and deep RFs (4 in area 1, 3 in area 2). Thirty-four neurons had no identifiable RF (4 in area 3b, 10 in area 1, and 20 in area 2). 3. The discharge of 185 of 359 neurons was significantly modulated during the presentation of one or both surfaces compared with the discharge at rest. Cells with a cutaneous RF that included part or all of the distal phalangeal pads of the digits used in the task (usually digits III and IV) were more likely to be modulated during surface presentation (132 of 179, 74%) than those with a cutaneous RF not in contact with the surfaces (24 of 104, 23%). The remaining neurons (mixed, deep, or no RF) were also infrequently modulated (29 of 76, 38%). 4. Of the 185 modulated units, 118 cells were classified as texture related because there was a significant difference in the discharge rate evoked by the smooth/rough and smooth surfaces. Cells with a cutaneous RF that included the digital pads in contact with the surfaces were frequently texture related (100 of 132, 76%). Texture sensitivity was less frequently observed in the remaining modulated neurons (18 of 53, 34%: cutaneous RF not in contact with the surfaces, deep RF, mixed cutaneous and deep RF, no identifiable RF). 5. Texture-related neurons were found in areas 3b, 1, and 2. Two patterns of texture-related responses were observed in the 100 cutaneous units with an RF in contact with the surfaces. Thirty-one units were classified as showing a phasic response at the time the digits encountered the leading edge of the rough half of the surface. Fifty-eight cells were classified as phasic-tonic (or sometimes tonic at the slowest motor speeds) because the response lasted for the duration of the presentation of the rough portion of the surface. The remaining 11 neurons could not be readily classified into one or the other category and, indeed, generally showed clear texture-related responses only at higher motor speeds (> 49 mm/s, 9 of 11). 6. Speed sensitivity was systematically evaluated in 41 of 100 texture-related units with a cutaneous RF in contact with the surfaces. The discharge of 66% of the units (27 of 41) varied significantly with the speed of surface presentation, with discharge increasing at higher speeds. Speed sensitivity was found in all three cytoarchitectonic areas (6 of 6 cells in area 3b, 11 of 22 in area 1, and 10 of 13 in area 2). 7. Contact force was also systematically monitored in these experiments (69 of 100 texture-related cells with a cutaneous RF in contact with the surfaces). Linear regression analyses indicated than 22% (15 of 69) of the texture-related units were sensitive to contact force (13

Animals↗

Visual evoked potentials with crossed asymmetry in incomplete congenital stationary night blindness.

PURPOSE: To investigate a proposed postretinal defect in patients with the incomplete form of congenital stationary night blindness (CSNB2) and to compare visual evoked potential (VEP) results with those found in various forms of albinism. METHODS: Visual evoked potentials were performed in 10 patients with a diagnosis of CSNB2, 10 subjects with albinism, and 17 normal subjects. Visual evoked potentials were elicited monocularly with diffuse flash stimulation. Scalp electrodes were placed over each hemisphere and referred to the forehead. Interhemispheric bipolar recordings were derived, and the correlation coefficient (CC) was calculated for various segments of the interhemispheric responses. RESULTS: A crossed visual evoked potential asymmetry pattern could be demonstrated in 9 of 10 patients with CSNB2. All subjects with albinism and none of the normal subjects showed the crossed asymmetry pattern. Statistical comparison of the CC computed for various segments of the interhemispheric response shows that the pattern of inversion in CSNB2 is more prominent in the 25 to 100 msec range (median CC, -0.37) and in the 175 to 250 msec range (CC, -0.27). In subjects with albinism, all segments show a negative CC (range, -0.46 to -0.60). In normal subjects, all segments are positively correlated (range, 0.36 to 0.66). CONCLUSIONS: Crossed visual evoked potential asymmetry was found in patients with CSNB2; therefore, excessive decussation, as demonstrated by this testing procedure, should not be considered as pathognomonic for albinism.

Adolescent↗

The electroretinographic diagnosis of the incomplete form of congenital stationary night blindness.

Fifteen patients with the incomplete form of congenital stationary night blindness (iCSNB) were reviewed to better characterize their electroretinographic (ERG) findings in view of differential diagnosis with other retinal conditions also presenting with negative bright-flash ERG responses. In all 15 patients, in dark-adapted conditions, the bright-flash ERG response had a normal a-wave followed by a subnormal b-wave. Oscillatory potentials (OPs) observed on the ascending limb of the b-wave, although delayed in implicit time, were of large amplitude. The response to a long-wavelength stimulus showed cone-related components and some well-delineated OPs. On the other hand, the photopically elicited cone responses were strongly abnormal, with a subnormal a-wave followed by a barely recordable b-wave. No OPs could be elicited under photopic conditions. The cone related components and the OP characteristics clearly distinguish iCSNB from the complete form of CSNB and other retinal conditions presenting with minimal fundus abnormalities but with negative bright-flash ERG responses, such as found in Duchenne muscular dystrophy and Aland Island eye disease. The severely abnormal post-synaptic components in the photopic recordings contrast with the well-differentiated cone activity evoked in scotopic conditions. We propose a cone system that does not respond optimally under the normal operating range (photopic) but rather under mesopic or scotopic conditions. In spite of the severe cone-ERG deficits, visual acuity was only slightly reduced. We propose that the defect, which interferes marginally with the neuronal flow of information, lies in the structures responsible for the building of the b-wave.

Adaptation, Ocular↗

Correlation between electroretinogram findings and molecular analysis in the Duchenne muscular dystrophy phenotype.

Fifteen consecutive patients with the Duchenne muscular dystrophy (DMD) phenotype were studied. Each patient was asked to undergo an ophthalmic examination, an electroretinogram (ERG), and to donate a blood sample for molecular diagnosis. All 15 patients had a normal ophthalmic examination. Electroretinography was successful in 14/15 patients. The ERG tracings were normal in seven patients, abnormal in seven, and unreliable in one. Blood for molecular analysis was obtained in 12/15 patients. In the seven patients with a normal ERG, five underwent molecular analysis, and in these five no deletion was detected in the dystrophin gene. In the seven patients with an abnormal ERG, six had molecular analysis available, and all six were found to have a deletion. These results suggest that patients with a classic DMD phenotype are genetically heterogeneous, and that this heterogeneity is reflected in the ERG.

Adolescent↗

Duchenne muscular dystrophy: negative scotopic bright-flash electroretinogram but not congenital stationary night blindness.

Patients with Duchenne muscular dystrophy (DMD) have recently been reported to have an abnormal scotopic electroretinogram (ERG) showing weak rod-related responses along with a negative configuration of the bright-flash response, which has been described as being similar to the one in congenital stationary night blindness (CSNB). We compared qualitatively and quantitatively the ERGs of 6 subjects with DMD, 10 subjects with the complete form of CSNB (cCSNB), 13 subjects with the incomplete form of CSNB (iCSNB) and 1 subject with complex glycerol kinase deficiency (CGKD). The rod-related activity and the bright-flash responses were abnormal and similar in all four groups. The cone-related activity, however, was within normal limits only in the DMD group; the b-wave was subnormal in CGKD, truncated in cCSNB and nearly absent in iCSNB. The electrophysiologic signature in DMD clearly distinguishes the retinal function of these patients from any other retinal condition so far described.

Adolescent↗

Duchenne muscular dystrophy: negative scotopic bright-flash electroretinogram and normal dark adaptation.

Recent reports indicate that most subjects with Duchenne muscular dystrophy (DMD) have a negative scotopic bright-flash electroretinogram (ERG). We performed dark adaptometry in seven patients with the DMD phenotype. Three patients had the negative bright-flash response combined with severely reduced rod-related activity; their mean psychophysical dark-adapted threshold was -5.5 +/- 0.3 (normally -5.4 +/- 0.3) cd.m-2. Two of the three patients were able to sit through the complete test: the profile of their dark adaptation curve was normal. The four other patients had a normal scotopic ERG, and their mean psychophysical dark-adapted threshold was -5.3 +/- 0.3 cd.m-2. One of the four was able to complete the entire procedure, and a normal profile was found. In patients with DMD the dark adaptation measurements are normal despite abnormalities in rod-related ERG activity.

Adolescent↗

Longitudinal study of the early electroretinographic changes in Alström's syndrome.

We obtained serial electroretinograms in four patients aged between 6 months and 5 years with Alström's syndrome and studied the early stages of the severe retinopathy that is characteristic of that disease. The weak electroretinographic signals found at age 6 months demonstrate a severe early cone dysfunction; one year later the cone activity is undetectable. The rod component of the electroretinogram is initially normal but can rapidly deteriorate to become undetectable as early as 5 years of age. These unusual electroretinographic findings are pathognomonic of Alström's syndrome and different from other cone-rod dystrophies or other syndromes with similar phenotypes such as Bardet-Biedl, Laurence-Moon, and Cohen syndromes.

Child, Preschool↗

Negative-configuration electroretinogram in Oregon eye disease. Consistent phenotype in Xp21 deletion syndrome.

OBJECTIVE: To determine whether abnormal configurations on electroretinogram were a consistent finding in patients with Xp21 deletion and to characterize the associated ophthalmologic phenotype. DESIGN: Case series. SETTING: University hospitals and eye institutes. PATIENTS: Five patients with complex glycerol kinase deficiency (Duchenne-type or Becker's muscular dystrophy, glycerol kinase deficiency, and congenital adrenal hypoplasia) and demonstrated chromosomal deletions at Xp21. Control patients were matched by age. MAIN OUTCOME MEASURES: Clinical information was obtained from medical records. Complete ophthalmologic examinations were performed. Electroretinography was performed using a Ganzfeld technique and chloral hydrate sedation. RESULTS: We report the clinical features and abnormal configurations on electroretinograms of five patients with complex glycerol kinase deficiency, including follow-up studies on a previously described patient. The original patient had ocular hypopigmentation; four, strabismus; two, myopia; three, astigmatism; and one, symptomatic night blindness. All had negative configurations on scotopic electroretinograms showing a reduced-amplitude B wave in the dark-adapted state. CONCLUSIONS: Our original report suggested a diagnosis of Aland Island eye disease, which appears to be an incomplete form of congenital stationary night blindness. Linkage data place Aland Island eye disease and congenital stationary night blindness at Xp11, whereas our patients had deletions at Xp21. The phenotype reported here may represent the effects of a single gene defect or the compound effects of the Xp21 contiguous gene syndrome (complex glycerol kinase deficiency). The phenotype is referred to as Oregon eye disease.

Adolescent↗