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Moisture content, thickness, and lesions of sole horn associated with thin soles in dairy cattle.

Thin soles and resulting lameness are often associated with increased culling rates in large dairy operations. The moisture content of the sole horn may cause the horn to be softer, thus predisposing it to an increased rate of wear. For that reason, the first objective of this study was to determine the moisture content of the sole horn of thin-soled cows. The second objective was to document physical changes of the claw associated with thin soles. Twenty-six adult Holstein cows showing signs of thin soles, and 16 normal cows were used in the study. Selection criteria for thin-soled cows included a dorsal wall length of less than 7.5 cm and the following clinical signs: painful gait, arched back, and specific leg lameness. Sixteen normal adult cows with dorsal wall length in excess of 7.5 cm were randomly selected. For thin-soled cows, the moisture contents of sole horn were 37.1 +/- 0.7% for front claws and 40.5 +/- 0.7% for rear claws. These values were significantly different from each other. Moisture contents of sole horn from normal cows were 31.08 +/- 0.93% for front claws and 33.1 +/- 0.9% for rear claws, which were significantly different from each other and different from thin-soled cows. Rear claws had higher moisture contents than front claws. Mean sole horn thickness was 4.23 +/- 0.18 mm for the rear lateral claws and 5.15 +/- 0.18 mm for the rear medial claws. The average dorsal wall lengths were 66.8 +/- 3.94 mm for rear lateral claws and 69.1 +/- 7.25 mm for rear medial claws. Thirty percent of rear feet with thin soles had pathological claw horn lesions including sole/white line separation (72%) and sole ulcers (28%). Of the affected claws, 13% had more than one lesion. Seventy-seven percent of claw lesions occurred in the lateral claw.

Animals↗

Approach to the diagnosis of thin basement membrane nephropathy in females with the use of antibodies to type IV collagen.

CONTEXT: Thin basement membrane nephropathy is recognized by a diffusely thin glomerular basement membrane (GBM) ultrastructurally. In contrast to Alport syndrome (AS), there is no GBM thickening, lamellation, or granular inclusions. Morphologically, there is overlap between thin basement membrane nephropathy and AS in female patients in whom there might be only thin GBM and no pathognomonic findings of AS. OBJECTIVE: To determine if the use of antibodies to collagen IV is helpful in making the distinction between thin basement membrane nephropathy and AS in female patients with primarily thin GBMs. DESIGN: We examined renal biopsies from 9 adult female patients with thin GBMs for the presence of alpha1, alpha3, alpha4, and alpha5 chains of type IV collagen by immunofluorescence. RESULTS: In 2 patients with segmental GBM staining, no suggestion for AS was found on physical examination or in their family history. In the remaining 7 patients with normal GBM staining, 4 had family members with end-stage renal disease of unknown etiology, raising the suspicion of X-linked or autosomal-recessive AS. Three patients were presumed to have thin basement membrane nephropathy. CONCLUSION: Segmental GBM staining for alpha3, alpha4, and alpha5 chains of type IV collagen raises the suspicion of AS in the presence of adequate controls and other supporting evidence. Normal GBM staining for alpha3, alpha4, and alpha5 chains of type IV collagen, however, does not exclude AS.

Adult↗

Effectiveness of thin-layer preparations vs. conventional Pap smears in a blinded, split-sample study. Extended cytologic evaluation.

OBJECTIVE: To further evaluate the effectiveness of the AutoCyte PREP thin-layer slide preparation (TriPath Imaging, Inc., Burlington, North Carolina) as compared to conventional Pap smears. STUDY DESIGN: A split-sample, blinded evaluation of matched thin-layer preparations and conventional smears from 2,438 patients was performed. This material was enriched by including 260 cases of high grade squamous intraepithelial lesions (HSILs) and cancer cases from an earlier study. Many of these cases were difficult to diagnose, containing very few abnormal cells on one or both matching slides. The preparations were evaluated multiple times by both thin-layer-inexperienced and -experienced cytology professionals to better compare performance related to preparation quality alone. RESULTS: The initial evaluations of the slides by personnel with only brief training in thin-layer interpretation demonstrated equivalent performance for the two preparations. The reevaluation study by cytology professionals with several months of thin-layer experience demonstrated a statistically significant improvement in detection of both LSIL and HSIL lesions using AutoCyte PREP slides. There was also a statistically significant improvement in the number of satisfactory samples using the AutoCyte PREP method. CONCLUSION: The study demonstrated that the AutoCyte PREP thin-layer slide preparation is at least equivalent to conventional Pap smears in the detection of LSIL and HSIL, even when evaluated by cytology professionals who have been newly trained in the thin-layer method and that, with increased experience, the thin-layer AutoCyte PREP slide preparation method showed a statistically significant improvement in disease detection.

Female↗

Papanicolaou and thin-layer cervical cytology with colposcopic biopsy control. A comparison.

OBJECTIVE: To compare the accuracy of conventional Papanicolaou and fluid-based, thin-layer cervical cytology. STUDY DESIGN: Cervical cytology was performed in duplicate on women who presented for cervical screening. Papanicolaou and thin-layer (ThinPrep, Cytyc Corp., Boxborough, Massachusetts) cytologic samples were collected simultaneously using a split-sample method. Cytologic slides were read and reported independently. Clinical follow-up was based on the most abnormal result. Colposcopy was performed as clinically indicated, and biopsy results were compared with cytologic diagnoses. RESULTS: Three thousand samples were compared. Papanicolaou and thin-layer results were significantly different (P = .0001), with identical diagnoses in 1,844 (61%) of patients. Eighty thin-layer (2.7%) and 177 Papanicolaou (5.9%) samples were read as limited or unsatisfactory (P < .0001). The rates of atypical squamous cells of undetermined significance (ASCUS) were not statistically different (P = .06). Thin-layer cytology was read more often as low grade squamous intraepithelial lesion (P = .001) or high grade squamous intraepithelial lesion (P = .006). Colposcopy with biopsy was performed on 291 patients. With ASCUS considered an abnormal result, thin-layer cytology was more sensitive (91% vs. 85%) but had lower positive predictive value (69% vs. 74%) than Papanicolaou cytology for the presence of cervical neoplasia. CONCLUSION: Papanicolaou and thin-layer cervical cytology yielded significantly different information. Thin-layer cytology yielded significantly fewer unsatisfactory results and was more sensitive for identifying cervical intraepithelial neoplasia.

Adult↗

Coronal multiplanar reconstruction view from whole lung thin-section CT by multidetector-row CT: determination of malignant or benign lesions and differential diagnosis in 68 cases of solitary pulmonary nodule.

PURPOSE: The purpose of this study was to evaluate the usefulness of the coronal multiplanar reconstruction (MPR) view in comparison with transverse helical thin-section CT for both the determination of malignant or benign lesions and the differential diagnosis of solitary pulmonary nodules. MATERIALS AND METHODS: Sixty-eight cases of pathologically proved solitary pulmonary nodule less than 3 cm in diameter were enrolled in this study. For the routine study, transverse helical thin-section CT (1.25 mm collimation, FOV 20 cm) covering the areas with solitary pulmonary nodules as well as whole lung helical thin-section CT (2.5 mm collimation, 1.25 mm reconstruction interval, FOV 34.5 cm, pitch 6:1, high-spatial frequency algorithm) were scanned with a multidetector-row CT (MDCT) scanner. From the whole lung thin-section CT data, coronal MPR views (2.5 mm slice thickness) were reconstructed on a workstation. ROC analysis was used for an observer performance study, in which three observers indicated their confidence level for the determination of malignant or benign lesion for the nodules by means of transverse thin-section CT and coronal MPR. In addition, the observers recorded appropriate disease entities as the final diagnosis of each case. Accuracies of the final diagnosis based on the two sets of images were compared with McNemer' s test. RESULTS: In terms of the determination of malignant or benign lesion, there was no significant difference between the two sets of images (coronal MPR and transverse thin-section CT; mean Az=0.853 and 0.854, respectively). In addition, accuracy of the final diagnosis based on coronal MPR views (74%) was almost equal to that based on transverse thin-section CT (71%) (p=0.3). CONCLUSIONS: The diagnostic efficacy of the coronal MPR view is comparable to that of transverse thin-section CT for the evaluation of solitary pulmonary nodules.

Adult↗

Drive for thinness in black and white preadolescent girls.

This study examined racial differences in drive for thinness, a motivational variable implicated in the etiology of eating disorders. Subjects included 613 black and white preadolescent girls from one of three National Heart, Lung, and Blood Institute (NHLBI) Growth and Health Study centers. Instruments included the Drive for Thinness Scale, a Criticism about Weight scale, the Self-Perception Profile for Children, a Sexual Maturation index, and 3-day food diaries. Black girls reported significantly greater drive for thinness than white girls. Drive for thinness was significantly associated with adiposity in both groups; additional predictors included criticism about weight for black girls and dissatisfaction with physical appearance for white girls. Correlations between drive for thinness and nutrient intakes were not significant. The finding of a greater drive for thinness among young black girls is provocative, given the higher prevalence of obesity and the lower prevalence of anorexia nervosa among black women. Longitudinal follow-up will examine the significance of drive for thinness in the development of weight and eating disorders in this cohort.

Black or African American↗

The influence of puberty and ethnicity on awareness and internalization of the thin ideal.

OBJECTIVE: This study examined the effects of puberty and ethnicity on awareness and internalization of the thin ideal. METHOD: Fourth and fifth-grade girls (N = 67) and their parents (N = 67) completed questionnaires assessing demographic variables, eating pathology, pubertal development, and awareness and internalization of the thin ideal. RESULTS: Girls in pre/early puberty did not differ significantly from girls in mid/late puberty on awareness of the thin ideal; however, more developed girls had significantly greater internalization of the thin ideal. Caucasian and non-Caucasian girls also did not differ on awareness of the thin ideal, but non-Caucasian girls reported greater internalization. No significant interactions between race and puberty existed on the dependent variables. DISCUSSION: Factors specifically related to puberty seem to be key to girls' development of disordered eating attitudes. The effect of puberty on increased internalization of the thin ideal cannot be explained by increased age or awareness of the thin ideal.

Adolescent↗

Effects of short-term overfeeding on hunger, satiety, and energy intake in thin and reduced-obese individuals.

BACKGROUND: Certain individuals appear to be resistant to weight gain in an obesigenic environment, yet the mechanisms for this adaptation are unclear. These individuals may sense positive energy balance more appropriately than those individuals prone to weight gain. RESEARCH METHODS AND PROCEDURES: Thirteen thin (7 women, 6 men) and 9 reduced-obese (5 women, 4 men) individuals were studied. Measures of intake were obtained using visual analog scales before and after each meal during eucaloric feeding and during three days of 50% overfeeding. Ad libitum energy intake was measured post-overfeeding. RESULTS: Overfeeding resulted in a significant reduction in pre-meal hunger in the thin (68+/-6 to 41+/-6 mm, p<0.0001) compared to the reduced-obese individuals (63+/-7 to 65+/-7 mm, p=0.67). There was a significantly greater increase (p=0.0016) in post-meal satiety scores during overfeeding in the thin (65+/-4 to 88+/-4 mm, p<0.0001) compared to the reduced-obese individuals (72+/-5 to 80+/-5 mm, p=0.04). Thin women had a 30% reduction in daily energy intake (1831+/-47 to 1273+/-79 kcal/day, p=0.005) after overfeeding while no difference was seen in the thin men or reduced-obese group. DISCUSSION: Thin individuals appear to sense positive energy balance more appropriately with changes in measures of intake than reduced-obese individuals. These findings are especially true in thin women.

Adaptation, Physiological↗

Body mass index in 7-9-y-old French children: frequency of obesity, overweight and thinness.

OBJECTIVE: To assess the frequency of different grades of nutritional status (obesity, overweight and thinness) in French children aged 7-9 y using four current definitions based on body mass index (BMI). METHODS: Data were collected in 2000 in a randomly selected sample of French children following the protocol recommended by the European Childhood Obesity Group (ECOG). After computing the BMI (weight/height squared), four references were used to define grades of nutritional status: (1) the French references to define thinness and overweight (3rd and 97th percentiles respectively); (2) the Must et al references to define thinness, overweight and obesity (5th, 85th and 95th percentiles respectively); (3) the International Obesity Task Force cut-offs to define overweight and obesity; and (4) the Center for Disease Control 2000 references to define thinness, overweight and obesity (5th, 85th and 95th percentiles respectively). RESULTS: Age and gender standardized frequencies were estimated in 1582 children. According to the French, Must et al, IOTF and CDC references, overweight (including obesity) affected 16.3, 23.9, 18.1 and 20.6% of children, respectively; obesity affected 9.3, 3.8 and 6.4% of children according to the Must, IOTF and CDC references, respectively. Thinness was present in 3.9, 6.0 and 6.0% of children according to the French, Must and CDC references. Whatever the definition, little difference was observed between sexes. Through age classes, as a rule, overweight and obesity tended to decrease while thinness tended to increase. CONCLUSION: The present study revealed an increasing prevalence of overweight in comparison with previous French data and a trend for increasing prevalence of thinness. The IOTF-based prevalence of overweight (including obesity) in 2000 in France was similar with the prevalence recorded in the late 1980s in the USA and the prevalence of obesity in 2000 in France was similar to the prevalence of obesity in the late 1970s in the USA. Data in France are comparable to those reported in other studies conducted in Western Europe. This study provides baseline information for analysis of time trends and for geographical comparisons.

Body Mass Index↗

Heart rate and systolic blood pressure variability: the impact of thinness and aging in human male subjects.

BACKGROUND: Life spans are steadily increasing in developing countries where 'thinness' is widely prevalent. However, the interaction of aging and thinness has been poorly studied in terms of its physiological consequences. OBJECTIVE: To determine the impact of aging and 'thinness' (body mass index (BMI) < 18.5 kg/m2) on resting heart rate and systolic blood pressure (SBP) variability indices in the frequency domain. SUBJECTS: Ninety seven healthy male subjects were divided into two age categories; young; 18-30 yrs and old superior 60 yrs. The subjects were further divided on the basis of BMI into young, thin (n=32), young, normal BMI (BMI 18.5-25, n = 27), old, thin (n = 15) and old, normal BMI (n = 23) groups. METHODS: Cardiac autonomic nerve function was determined using heart rate variability indices in the frequency domain (low frequency, cardiac sympathetic 0.04- 0.15 Hz; high frequency, cardiac parasympathetic 0.15-0.4 Hz). Vasomotor sympathetic activity was determined from the low frequency component of SBP variability. Baroreflex sensitivity was determined from the spectral power of both RR variability and SBP variability between 0.07 to 0.14 Hz. RESULTS: Thinness was associated with a reduction in the absolute total, low and high frequency heart rate power spectrum as compared to individuals of normal BMI, but this difference was only apparent in young adults (P < 0.05) and not in older subjects. The age related decline in heart rate variability (absolute units) was apparent for subjects of both low and normal BMI (P < 0.05). There were no differences in SBP variability either with age or BMI. CONCLUSIONS: The data suggest that aging has a pronounced effect on heart rate variability, which may mask differences in heart rate variability related to thinness.

Adolescent↗

Thin foil preparation of metal particles in brittle ceramic matrices.

Thin foil preparation for metal particles in brittle ceramic matrices is particularly difficult, due to differential thinning of the component materials. This paper considers the preparation of thin foils of stony meteorites. Stony meteorites have a composite nature consisting of small metal particles (< 200 microns) embedded in an inherently brittle silicate matrix. Specimens of this type are difficult to prepare as thin foils because the area of interest is limited to specific regions within the metal particles. The problems of thin foil preparation of metal particles in the stony meteorites were overcome by developing a technique involving electrochemical polishing prior to ion beam thinning. The metal particles were first separated from the silicate matrix and then embedded in Epo-Tek H20E silver epoxy. This specific epoxy was selected for its good electron conductivity which allowed the specimen to be thinned by the electrochemical polishing technique. Specimens prepared by this technique have permitted the direct observation and characterization of the metallic phases in stony meteorites.

Ceramics↗

Distribution and orientation of rhodamine-phalloidin bound to thin filaments in skeletal and cardiac myofibrils.

Phalloidin staining of muscle does not reflect the known disposition of sarcomeric thin filaments. Quantitative image analysis and steady-state fluorescence polarization microscopy are used to measure the local intensity and orientation of tetramethyl rhodamine-labeled phalloidin (TR-phalloidin) in skinned myofibrils. TR-phalloidin staining of isolated skeletal myofibrils labeled while in rigor reveals fluorescence that is brighter at the pointed ends of the thin filaments and Z lines than it is in the middle of the filaments. In cardiac myofibrils, phalloidin staining is uniform along the lengths of the thin filaments in both relaxed and rigor myofibrils, except in 0.2-micron dark areas on either side of the Z line. Extraction of myosin or tropomyosin-troponin molecules does not change the nonuniform staining. To test whether long-term storage in glycerol changes the binding of phalloidin to thin filaments in myofibrils, minimally permeabilized (briefly skinned) myofibrils, or myofibrils stored in glycerol for at least 7 days (glycerol extraction) were compared. TR-phalloidin was well ordered throughout the sarcomere in briefly skinned skeletal and cardiac myofibrils, but TR-phalloidin bound to the Z line and pointed ends of thin filaments was randomly oriented in glycerol-extracted myofibrils, suggesting that the ends of the thin filaments become disordered after glycerol extraction. In relaxed skeletal myofibrils with sarcomere lengths greater than 3.0 microns, staining was nearly uniform all along the actin filaments. Exogeneous bare actin filaments polymerized from the Z line (Sanger et al., 1984: J. Cell Biol. 98:825-833) in and along the myofibril bind rhodamine phalloidin uniformly. Our results support the hypothesis that nebulin can block the binding of phalloidin to actin in skeletal myofibrils and nebulette can block phalloidin binding to cardiac thin filaments.

Actin Cytoskeleton↗

Early segmental thinning of the left ventricular wall following regional ischemia.

To explore possible mechanisms of left ventricular early segmental relaxation, complete occlusion of the left anterior descending coronary artery (LAD) was produced in seven open-chest dogs and partial occlusion of the LAD was produced in six open-chest anesthetized dogs. Regional wall thickness was measured both in an ischemic and a normally perfused zone using implanted ultrasonic crystals. Two to three seconds following complete LAD occlusion, thinning of the ischemic wall occurred prematurely during isovolumic relaxation. The extent of premature thinning became more prominent 5 to 10 sec following LAD occlusion. Early thinning of the ischemic wall preceded thinning of the normally perfused wall by 110 +/- 10 msec. Partial occlusion of the LAD produced a 33 +/- 6% reduction of coronary flow and a 23 +/- 4% reduction of systolic wall thickening in the ischemic region. Systolic thickening of the nonischemic wall was unchanged relative to the preocclusion period. Premature early thinning of the mildly ischemic wall preceded thinning of the normally perfused segment by 90 +/- 8 msec. The observation that ischemia can produce segmental early thinning of the ventricular wall may have implications in understanding the mechanism of the angiographic observation of the segmental early relaxation phenomena in patients with coronary artery disease.

Animals↗

Adverse effects of social pressure to be thin on young women: an experimental investigation of the effects of "fat talk".

OBJECTIVE: Experiments have found that pressure to be thin from the media promotes body dissatisfaction and negative affect, but the effects of social pressure to be thin have not been examined experimentally. Thus, this study tested whether social pressure to be thin fosters body dissatisfaction and negative affect. METHOD: Young women (N = 120) were randomly assigned to a condition wherein an ultra-thin confederate complained about how fat she felt and voiced intentions to lose weight or a control condition wherein she discussed a neutral topic. RESULTS: Exposure to social pressure to be thin resulted in increased body dissatisfaction but not negative affect. The effects were not moderated by initial thin-ideal internalization, body dissatisfaction, or social support. DISCUSSION: Results support the assertion that peer pressure to be thin promotes body dissatisfaction but suggest that this factor may not contribute to negative affect.

Adolescent↗

Preparation of (InGa)As/GaAs multilayered materials for TEM by one side non-rotation ion beam thinning.

A technique is described for the preparation of thin specimens for transmission electron microscopy (TEM) of (InGa)As/GaAs multilayered materials. In this technique, a shielding method is used for selective-area perforation by ion beam thinning. Thin cross-sectional specimen slices are mechanically pre-thinned to about 30 microns and then thinned by ion sputtering from one side of the specimen at a time without rotation of the specimen stage. No direct ion sputtering occurs at the growth surface of the specimen so that a specimen with thin areas containing the desired near-surface structures can be obtained. The recipe for this technique is given in detail. A patterning method for increasing the size of the thin area for TEM investigation is also described. It is shown that a smooth surface can be obtained by sputtering without rotating the stage if obstacles that produce redeposits onto the sputtered surface are removed.

Arsenic↗

States of thin filament regulatory proteins as revealed by combined cross-linking/X-ray diffraction techniques.

The regulatory protein system in the skeletal muscle thin filaments is known to exhibit three discrete states, called "off" or "blocked" (no Ca2+), "on" or "closed" (with Ca2+ alone) and "potentiated" or "open" (with strongly bound myosin head) states. Biochemical studies have shown that only weak interactions with myosin are allowed in the second state. Characterization of each state is often difficult, because the equilibria among these states are readily shifted by experimental conditions. To overcome this problem, we chemically cross-linked the skeletal muscle thin filament in the three states with the zero-length cross-linker 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), in overstretched muscle fibers. The state of the regulatory proteins was monitored by measuring the intensity of the second actin layer-line (2nd LL) reflection in X-ray diffraction patterns. Structurally, the thin filaments cross-linked in the three states exhibited three corresponding discrete levels of 2nd LL intensities, which were not Ca2+-sensitive any more. Functionally, the thin filament cross-linked in the "off-blocked" state inhibited strong interaction with myosin head (subgfragment-1 or S1). The thin filament cross-linked in the "potentiated-open" state allowed strong interaction and full ATPase activity of S1 as described previously. The thin filament cross-linked in the "on-closed" state allowed strong interactions with S1 and actin-activated ATPase without enhancing the 2nd LL to the level of "potentiated-open" state, contrary to the expectations from the biochemical studies. The results demonstrate the potential of EDC as a tool for studying the states of calcium regulation, and the apparent uncoupling between the 2nd LL intensity and the function provides a new insight into the mechanism of thin filament regulation.

Actins↗

Filamin and gelsolin influence Ca(2+)-sensitivity of smooth muscle thin filaments.

Sheep aorta thin filaments were prepared by ultracentrifugation of an ATP-containing extract in the presence of different concentrations of ethanediol. Thin filaments prepared without ethanediol contained small quantities of tropomyosin (0.027 Tm:actin) and caldesmon (0.017 CD:actin) and activated the MgATPase of skeletal myosin independently of Ca2+. Ultracentrifugation in the presence of 10-20% ethanediol resulted in preparation of thin filaments with increased content of tropomyosin (0.17 Tm:actin) and caldesmon (0.04 CD:actin). These thin filaments possessed high Ca(2+)-sensitivity in activation of skeletal muscle myosin ATPase. Besides actin, tropomyosin and caldesmon, thin filaments contained gelsolin and filamin. Gelsolin content (0.007 gelsolin:actin) was independent of the presence of ethanediol. The filamin content decreased from 0.015 to 0.007 mol:mol actin when the ethanediol concentration was increased from 0 to 20%, and was negatively correlated with the Ca2+ sensitivity of thin filaments. In a reconstituted system, pure filamin or gelsolin affected caldesmon regulation of actomyosin ATPase. Gelsolin (0.01:actin) reduced the inhibition of actomyosin ATPase caused by caldesmon and increased the potency of Ca(2+)-calmodulin in reversing this inhibition. Filamin (0.007:actin) also decreased the inhibitory action of caldesmon on actin-activated myosin ATPase and also potentiated the reversal of this inhibition by calmodulin. We conclude that minor components of smooth muscle thin filaments (gelsolin and filamin) significantly modify caldesmon mediated regulation of actomyosin ATPase. We suggest a tropomyosin-mediated mechanism by which filamin or gelsolin could exert similar effects.

Actin Cytoskeleton↗

Structural and functional reconstitution of thin filaments in skeletal muscle.

Thin filaments were reconstituted by incorporating exogenous actin, tropomyosin and troponin into glycerinated skeletal muscle fibres or myofibrils. Firstly, thin filaments except short fragments at the Z line were selectively removed by treatment with plasma gelsolin, an actin severing protein. As a result, the fibres (or fibrils) lost the ability to generate active tension. Next, actin filaments were reconstituted by adding purified G-actin which polymerizes onto the actin fragments which remained at the Z line. Rhodamine phalloidin staining of myofibrils showed that exogenous actin was incorporated into the position where the intrinsic thin filaments located. Thin section electron micrographs of fibres showed that reconstituted actin filaments ran from the Z line to the inside of the A band, with some reaching the H zone. The number density of reconstituted actin filaments in the A band was about 20% of that found in intact fibres. The actin filament-reconstituted fibres (or fibrils) generated active tension in a Ca(2+)-insensitive manner and the tension was reversibly suppressed by 2,3-butanedione 2-monoxime. The recovered active tension was about 20% of tension developed by intact fibres. These results indicate that reconstituted actin filaments bear active tension similar to that borne by intact thin filaments. Thin filament-reconstituted fibres, which were prepared by adding purified tropomyosin-troponin complexes into actin filament-reconstituted fibres, showed Ca(2+)-sensitive tension generation. The maximum tension generated was not affected by the presence of tropomyosin and troponin. SDS-PAGE analysis showed that more than 25% of actin and 20% of tropomyosin and troponin was incorporated into the reconstituted fibres. These results indicate that the structure and function of thin filaments are substantially reconstituted by self-assembly of actin, tropomyosin and troponin. The reconstituted fibres and fibrils will be useful for studying the molecular mechanism of muscle contraction and its regulation.

Actin Cytoskeleton↗