Far-infrared emission by resonant-polaron effects in narrow-band-gap Hg1-xCdxTe.
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Journal Article↗
Biomedical subjects
F Fuchs
Publications and source records attributed to F Fuchs.
Heterogeneity of capsid proteins of echovirus type 25 wild-type strain and prototype strain, studied by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting.
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting were used to compare the capsid proteins of 19 antigenic variants of echovirus type 25 wild-type strains isolated in France between 1976 and 1987 with those of the prototype JV-4 reference strain isolated in 1957. Immunoblots were developed by using polyclonal sera from rabbits and mice immunized with the reference strain. Immunoblotting patterns revealed reactivity only against viral protein VP1 for sera from both animals. Comparative immunoblotting patterns showed differences in the electrophoretic mobilities of viral protein VP1, especially for the Montpellier 76.1262 wild-type strain. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of [35S]methioinine-labeled viral polypeptides revealed that the two variant strains, Montpellier 76.1262 and Thionville 86.222, exhibited significant and reproducible shifts in the relative mobilities of VP1 and VP3 and, to a lesser extent, in those of VP0 and VP2. The relative mobility of VP4 seemed very similar for the JV-4 reference strain and the two variants. Interestingly, the structural differences in VP1 and VP3 of Montpellier 76.1262 were not correlated with the pattern of neutralization by monoclonal antibodies, unlike in our previous study, in which this strain differed from the prototype strain in only two epitopes. We concluded that, in addition to the heterogeneity of their biological and antigenic properties that we observed previously, echovirus type 25 wild-type strains may exhibit differences in their structural proteins.
Animals↗
Force, length, and Ca(2+)-troponin C affinity in skeletal muscle.
On the basis of isotopic methods it has been found that force generation promotes increased Ca2+ binding to troponin C in cardiac muscle [P. Hofmann and F. Fuchs. Am. J. Physiol. 253 (Cell Physiol. 22): C541-C546, 1987] but not in skeletal muscle (J. Muscle Res. Cell Motil. 6: 477, 1985). However, studies with skinned rabbit psoas muscle fibers containing substituted fluorescent troponin C analogues indicate that force-generating cross bridges do promote increased Ca2+ binding in skeletal muscle (K. Güth and J. D. Potter. J. Biol. Chem. 262: 13627-13635, 1987). We have reexamined this question using a modified contraction-relaxation protocol in which Ca2+ binding to detergent-treated rabbit psoas fibers was measured either during steady-state force development or after relaxation was induced by one of two myosin ATPase inhibitors, vanadate or 2,3-butanedione monoxime. A standard double-isotope technique was used to measure Ca2+ binding. Another set of experiments was done in which force was reduced by releasing muscle fibers from sarcomere lengths of 2.4-2.6 microns to 1.5-1.7 microns, and bound Ca2+ was determined either before or after the release. No statistically significant effect of force generation or sarcomere length on Ca(2+)-troponin C affinity was observed. Thus the discrepancy remains between results obtained with isotopic and fluorescence methods. It is possible that in skinned fibers emission from fluorescence probes is more closely related to protein-protein interactions than to the amount of Ca2+ bound to troponin C.
Animals↗
Impact on routine diagnosis of echovirus infections of intratypic differentiation and antigenic variation in echovirus type 25 studied by using monoclonal antibodies.
We studied the biological and antigenic properties of wild strains of echovirus type 25 isolated in France between 1982 and 1987 and compared them with the JV-4 prototype strains isolated in 1957. The wild strains differed from the prototype strain in their cellular tropism. The prototype strain grew readily in five cell lines (MRC5, MA 104, Vero, BGM, and HT 29-18), while for wild strains MRC5 and HT 29-18 cells were the most sensitive and supported growth to high titres (between 4.5 and 7.4 50% tissue culture infective doses per 0.05 ml). Plaques produced by wild strains were larger (6.05 +/- 0.94 mm in diameter [mean +/- standard deviation]) than those of the prototype strain (2.3 +/- 0.97 mm in diameter) and heterogeneous, even after cloning by three terminal dilution passages, which suggested heterogeneous virus populations. Virus neutralization with polyclonal monovalent sera showed that wild strains were significantly less neutralized by two reference immune sera than the prototype strain was. Monoclonal antibodies were raised against the echovirus type 25 JV-4 prototype strain. Nine clones with neutralizing activity were identified. Heterologous neutralizations of 14 clinical isolates revealed highly conserved, moderately conserved, and poorly conserved epitopes. The natural isolates differed from the prototype strain in two to four epitopes and can be classified into four different groups. We concluded that echovirus type 25, like coxsackie- and polioviruses, consists of heterogeneous viral populations with respect to biological and antigenic properties. In term of viral diagnosis, it may become increasingly difficult to identify recently isolated strains because of their antigenic variation.
Antibodies, Monoclonal↗
Specific detection of enteroviruses in clinical samples by molecular hybridization using poliovirus subgenomic riboprobes.
Enteroviruses were specifically detected in crude clinical specimens or in cell cultures in which the viruses were amplified by dot hybridization by using poliovirus type 1-derived, subgenomic radiolabeled cRNA probes (riboprobes). The sensitivity of this test varied from 2.5 to 33%, when clinical specimens without cell culture were examined, and was about 85% in cell culture lysates. The specificity of the test was 90 to 100%. The riboprobe corresponding to the 5'-noncoding sequence specifically detected the majority of enteroviruses (56 of 57 tested); the riboprobe derived from the VPI capsid region hybridized with the three poliovirus serotypes and with some coxsackieviruses type A and with echovirus type 7. Echovirus 22 did not hybridize with any riboprobe. In stool specimens, nasal aspirates, and cerebrospinal fluids from patients with meningitis, only one type of virus was identified in different clinical samples from the same patient by the seroneutralization test. Hybridization allowed the detection of enteroviral RNAs easily in stool specimens and nasal aspirates but with a low efficiency in cerebrospinal fluids without amplification of the viruses in cell cultures.
Capsid↗
The reactivity of sulfhydryl groups of bovine cardiac troponin C.
Bovine cardiac troponin C (cTnC) contains 2 cysteine residues, Cys-35 located in the nonfunctional Ca2+-binding loop I and Cys-84 in the N-terminal segment of the central helix. We have studied the reactivity of Cys residues in cTnC with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM). The latter compound fluoresces only when reacted with the protein. The reaction with DTNB followed second order kinetics with respect to DTNB, the rate constants being 3.37 s-1 M-1 and 1.82 s-1 M-1 in the presence and absence of Ca2+, respectively. These rates are much slower than the rate of reaction with Cys-98 of skeletal TnC (sTnC) or with the urea-denatured cTnC, indicating that both Cys residues are partly buried within the structure of the protein. The increase in reactivity was induced by binding of Ca2+ to the single low affinity Ca2+ binding site (site II). The fluorescence increase upon reaction of cTnC with CPM in the absence of Ca2+ could be fitted with a single exponential equation indicating that both cysteine residues are equally available to the reagent. The reaction in the presence of Ca2+ was biphasic. Analysis of CNBr fragments of cTnC labeled with CPM under various conditions indicated that in the presence of Ca2+ the reactivity of Cys-84 is increased while that of Cys-35 is slightly decreased. This finding is consistent with the model of Herzberg et al. (Herzberg, O., Moult, J., and James, M. N. G. (1986) J. Biol. Chem. 261, 2638-2644) and the data of Ingraham and Hodges (Ingraham, R. H., and Hodges, R. S. (1988) Biochemistry 27, 5891-5898), suggesting that the Ca2+-induced conformational change in the N-terminal half of TnC involves separation of the helix C from the central helix, thereby increasing the accessibility of Cys-84. The slow overall kinetics, however, indicates that the structure in the vicinity of Cys residues is relatively compact regardless of Ca2+. We interpret the increase in reactivity towards CPM as consistent with a Ca2+-induced exposure of a hydrophobic pocket in the vicinity of Cys-84.
Amino Acids↗
Optical properties of the SbGa heteroantisite defect in GaAs:Sb.
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Journal Article↗
Nutrient intake and vitamin status of healthy French vegetarians and nonvegetarians.
The status of thiamin, riboflavin, folate, and vitamins B-6, B-12, C, A, D, and E was investigated in 37 middle-aged and healthy French vegetarians by means of a dietary survey and biochemical studies. Values were compared with those of a group of nonvegetarians. Unsatisfactory intakes of vitamin B-6 were observed: vitamin B-6 intake as a percentage of the French Recommended Dietary Allowances was approximately 66% for vegetarians and approximately 58% for nonvegetarians. Vegetarians had a higher mean intake of thiamin, riboflavin, and vitamins C, A, D, and E than did nonvegetarians. Vegetarians did not have a higher risk rate for a biochemical vitamin deficiency of thiamin, riboflavin, folates, and vitamins B-6, C, A, and E than the nonvegetarians. The percentage of subjects assessed as abnormal by blood vitamin concentrations was higher in vegetarians for vitamin B-12 (serum vitamin B-12) and vitamin D, which indicated a higher risk for a deficiency of vitamins B-12 and D in this group.
Adult↗
Vitamin status of young athletes including the effects of supplementation.
Vitamin B1, B2, B6, C, A, and E status was assessed by combining determination of dietary intake with biochemical indices in 55 young male athletes. When compared with 20 matched sedentary controls, a similar number of individuals with biochemical deficiency was found in both groups for vitamins B1, B6, and E despite a higher mean dietary intake of these three vitamins in young athletes. Markedly reduced levels of pyridoxal 5'-phosphate, the biologically active form of the vitamin B6 compounds, were found in both populations. Supplementation administered for one month improved the vitamin status of the control subjects but not entirely those of the young athletes.
Adult↗
Bound calcium and force development in skinned cardiac muscle bundles: effect of sarcomere length.
There is evidence that the steep ascending limb of the force-length curve in cardiac muscle (Frank-Starling relation) is based on a length-dependence of myofilament Ca2+ sensitivity. Previous work from this laboratory has indicated that in the sarcomere length range corresponding to the ascending limb of the cardiac force length curve (1.7 to 2.3 microns) the Ca2+-troponin C affinity is length-dependent. In this study Ca2+ binding to chemically skinned bovine cardiac muscle bundles was measured during ATP-induced force generation with fiber bundles having sarcomere lengths of 2.2 to 2.4 microns and 1.6 to 1.8 microns. A double isotope technique was used to make concurrent determinations of the force-pCa and bound Ca2+-pCa relationships. At the longer sarcomere lengths the fibers bound, at saturation, an amount of Ca2+ equivalent to approximately 3 mol Ca2+/mol troponin C. Force development appeared to be coupled to titration of the single, low-affinity Ca2+-specific site. In the pCa range 7.0 to 6.0 sarcomere length had no effect on Ca2+ binding. In the pCa range 6.0 to 5.0, in which force increased steeply, there was, in addition to a decreased relative force, a significant reduction in bound Ca2+ at the shorter sarcomere length. Thus sarcomere length appears to influence the Ca2+ binding properties of the regulatory site on troponin C. These data provide direct evidence that length-dependent modulation of Ca2+-troponin C affinity may make a major contribution to the force-length relationship in cardiac muscle.
Animals↗
Fatty acid composition of serum lipids and its relation to diet in an elderly institutionalized population.
This study was conducted to determine the nutritional status with respect to fatty acids of 53 institutionalized elderly subjects (group A). Seven-day food records and biochemical determinations were used. The same protocol was applied to 25 healthy young adults (group B). The 1981 French Recommended Dietary Allowances were used to assess adequacy of intake. Total fat intake was found to be too high in both groups. Higher intakes of animal fats and saturated fatty acids and lower intakes of polyunsaturated fatty acids and linoleic acid were documented in group A than in group B. Elderly subjects exhibited decreased 18:2 omega 6 and 20:4 omega 6 in serum phospholipids and triglycerides. Other indicators of essential fatty acid status in group A differed from group B in the direction of deficiency (double-bond index [DBI], 0.93 +/- 0.01 vs 1.13 +/- 0.04, p less than 0.001; total omega 6, 25.07 +/- 0.46% vs 32.49 +/- 0.89%, p less than 0.001). These findings indicate that institutionalized elderly people are deficient in circulating essential fatty acids, which appears to be caused by both diet and metabolism.
Adult↗
Effect of length and cross-bridge attachment on Ca2+ binding to cardiac troponin C.
The sensitivity of skinned cardiac muscle bundles to Ca2+ is a function of sarcomere length. Ca2+ sensitivity is increased as fiber length is extended along the ascending limb of the force-length curve and it has been suggested that this phenomenon makes a major contribution to the steep force-length relationship that exists in living cardiac muscle. To gain greater insight into the mechanism behind the length dependence of Ca2+ sensitivity isotopic measurements of Ca2+ binding to detergent-extracted bovine, ventricular muscle bundles were made under conditions in which troponin C was the only major Ca2+ binding species. Experiments were designed to determine whether 1) Ca2+-troponin C affinity varies in the sarcomere length range corresponding to the ascending limb of the force-length curve, and 2) Ca2+ binding correlates with length per se or with changes in the number of length-dependent cross-bridge attachments. Measurements were made of Ca2+ binding in the rigor and relaxed states. The latter state was produced by suppressing actin-myosin interaction with the phosphate analogue, sodium vanadate. After vanadate treatment it is possible to obtain a complete Ca2+ saturation curve in the presence of physiological MgATP concentrations and at constant sarcomere length. The results show that the binding of Ca2+ to the regulatory site of cardiac troponin C is length dependent but this length dependence is actually a dependence on the number of attached cross bridges.
Adenosine Triphosphatases↗
Evidence for a force-dependent component of calcium binding to cardiac troponin C.
The duration of activation in cardiac muscle is a function of the load. On the basis of studies of Ca2+ transients in muscles subjected to quick release, it has been suggested that force or shortening-mediated changes in Ca2+-troponin C affinity may provide a mechanism for a contraction-activation feedback. This study was designed to test the hypothesis that the formation of force-generating complexes between actin and myosin enhances the affinity of cardiac troponin C for Ca2+. This was done by first establishing the normal relationship between Ca2+ binding and force development in chemically skinned bovine ventricular muscle bundles and then comparing the Ca2+-saturation curves obtained with relaxed and contracting muscle bundles. A double isotope technique was used to measure Ca2+ binding during ATP-induced force generation and during relaxation maintained by the phosphate analogue vanadate. The results showed that the generation of force was associated with an enhanced binding of Ca2+ to the Ca2+-specific regulatory site of cardiac troponin C. These data provide direct evidence that feedback between force and activation in the heart may be mediated by the Ca2+-regulatory site of troponin C.
Adenosine Triphosphate↗
[Sexual activity and pregnancy].
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Female↗
The binding of calcium to detergent-extracted rabbit psoas muscle fibres during relaxation and force generation.
Rigor complexes between actin and myosin have been shown to cause increased binding of Ca2+ to troponin C. A similar effect of force-generating crossbridges has been suggested as an explanation for the coupling between load and activation which has been observed in skeletal and cardiac muscle. The goal of this study was to test the hypothesis that Ca2+-troponin affinity during crossbridge cycling is load-dependent. Ca2+-binding to detergent-extracted rabbit psoas fibres was measured during ATP-induced force generation and in the relaxed state. To compare Ca2+ binding in the latter two states it was necessary to establish conditions in which ATP-induced force could be regulated independently of free Ca2+ concentration. Such conditions were obtained by the use of either the ATPase inhibitor sodium vanadate or the substitution of MgITP for MgATP as an energy source. This study showed that in the presence of MgATP (or MgITP) the amount of Ca2+ bound to the myofilaments at a given free Ca2+ concentration was independent of the force generated. Thus force per se is not a determinant of Ca2+-troponin affinity.
Animals↗
Oxytocin receptors in nonpregnant human uterus.
The concentration and distribution of specific [3H]oxytocin-binding sites in the nonpregnant human uterus were studied. Oxytocin was bound with an apparent Kd of about 1 nM in the crude membrane fractions of the fundus and corpus. A second class of sites with lower affinity and higher capacity for oxytocin also was found. Consistent with its high uterotonic potency in nonpregnant uteri, arginine vasopressin was as effective as oxytocin in inhibiting [3H]oxytocin binding to corpus and fundus membrane preparations. The concentrations of high affinity binding sites in fundus and corpus were similar and were significantly higher than those of membrane fractions from isthmus or Fallopian tubes. The lowest concentration of sites was found in the cervix. Endometrial membrane preparations contained oxytocin-binding sites in about the same concentration as that in the myometrium. The concentrations of oxytocin receptors in all parts of the nonpregnant uterus were somewhat higher in the luteal phase than in the follicular phase. Concentrations were lowest in postmenopausal uteri. The concentrations of oxytocin receptors in nonpregnant uteri were 50-100 times lower than those in uteri near the end of gestation. These differences correspond to the differences in sensitivity to oxytocin between luteal phase and follicular phase and between the nonpregnant and late term uterus. These findings add support to the evidence that the binding sites for oxytocin represent true oxytocin receptors.
Adult↗
The future of genetic amniocentesis.
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Amniocentesis↗