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At least 19 recordsLinked to original sources
PubMed2002

Ca2+- and S1-induced conformational changes of reconstituted skeletal muscle thin filaments observed by fluorescence energy transfer spectroscopy: structural evidence for three States of thin filament.

Rabbit skeletal muscle alpha-tropomyosin (Tm) and the deletion mutant (D234Tm) in which internal actin-binding pseudo-repeats 2, 3, and 4 are missing [Landis et al. (1997) J. Biol. Chem. 272, 14051-14056] were used to investigate the interaction between actin and tropomyosin or actin and troponin (Tn) by means of fluorescence resonance energy transfer (FRET). FRET between Cys-190 of D234Tm and Gln-41 or Cys-374 of actin did not cause any significant Ca2+-induced movement of D234Tm, as reported previously for native Tm [Miki et al. (1998) J. Biochem. 123, 1104-1111]. FRET did not show any significant S1-induced movement of Tm and D234Tm on thin filaments either. The distances between Cys-133 of TnI, and Gln-41 and Cys-374 of actin on thin filaments reconstituted with D234Tm (mutant thin filaments) were almost the same as those on thin filaments with native Tm (wild-type thin filaments) in the absence of Ca2+. Upon binding of Ca2+ to TnC, these distances on mutant thin filaments increased by approximately 10 A in the same way as on wild-type thin filaments, which corresponds to a Ca2+-induced conformational change of thin filaments [Miki et al. (1998) J. Biochem. 123, 324-331]. The rigor binding of myosin subfragment 1 (S1) further increased these distances by approximately 7 A on both wild-type and mutant thin filaments when the thin filaments were fully decorated with S1. This indicates that a further conformational change on thin filaments was induced by S1 rigor-binding (S1-induced or open state). Plots of the extent of S1-induced conformational change vs. molar ratio of S1 to actin showed that the curve for wild-type thin filaments is hyperbolic, whereas that for mutant thin filaments is sigmoidal. This suggests that the transition to the S1-induced state on mutant thin filaments is depressed with a low population of rigor S1. In the absence of Ca2+, the distance also increased on both wild-type and mutant thin filaments close to the level in the presence of Ca2+ as the molar ratio of S1 to actin increased up to 1. The curves are sigmoidal for both wild-type and mutant thin filaments. The addition of ATP completely reversed the changes in FRET induced by rigor S1 binding. For mutant thin filaments, the transition from the closed state to the open state in the presence of ATP is strongly depressed, which results in the inhibition of acto-myosin ATPase even in the presence of Ca2+. The present FRET measurements provide structural evidence for three states of thin filaments (relaxed, Ca2+-induced or closed, and S1-induced or open states) for the regulation mechanism of skeletal muscle contraction.

Actin Cytoskeleton↗
PubMed2000

Pre-operative endometrial thinning agents before hysteroscopic surgery for heavy menstrual bleeding.

BACKGROUND: Menorrhagia is one of the most common reasons for pre-menopausal women to be referred to a gynaecologist. Although medical therapy is generally the first approach, many will eventually require or request a hysterectomy. Hysterectomy is associated with a significant in-patient hospital stay and a period of convalescence that makes it an unattractive and unnecessarily invasive option for many women. Hysteroscopic endometrial ablation or resection offers a day-case surgical alternative to hysterectomy for these women. It is also a cheaper procedure than hysterectomy. Complete endometrial removal or destruction is one of the most important determinants of treatment success. Therefore surgery will be most effective if undertaken when endometrial thickness is less than 4mm, in the immediate post-menstrual phase, however there are often difficulties in reliably arranging surgery for this time. The other option is the use of hormonal agents which induce endometrial thinning or atrophy prior to surgery. The most commonly evaluated agents have been goserelin (a GnRH analogue) and danazol. Progestogens and other GnRH analogues have also been studied although less data are available. It has been suggested that the use of these agents, particularly GnRH analogues, will reduce operating time, improve the intra-uterine operating environment, and reduce distension medium absorption (this is the fluid used to distend the uterine cavity during surgery). OBJECTIVES: To investigate the effectiveness of gonadotrophin-releasing hormone (GnRH) analogues, danazol, and progestogens, when used for endometrial thinning prior to hysteroscopic surgery for menorrhagia, in improving the intra-uterine operating environment and treatment outcome after surgery. SEARCH STRATEGY: The Menstrual Disorders and Subfertility Group search strategy (see Review Group details) was used to identify randomised trials that had compared the use of these drugs with either each other, or placebo, or no pre-operative treatment. SELECTION CRITERIA: Trials were included if they compared the effects of these agents with each other, or with placebo or no treatment on relevant intra-operative and post-operative treatment outcomes. Only randomised studies were included in this review. DATA COLLECTION AND ANALYSIS: Eight studies met the inclusion criteria for this review. Four studies compared goserelin (a GnRH analogue) with no treatment or placebo. Three studies compared goserelin with danazol. One study compared progestogens, danazol and triptorelin (a GnRH analogue) with no treatment. Data was extracted independently by two reviewers. A third reviewer checked data extraction for accuracy and wrote to authors where relevant data was missing or unclear. Intra-operative parameters included endometrial thickness, duration of surgery, ease of surgery, distension medium absorption and complication rate. Post-operative outcomes compared were the proportion of women with amenorrhoea, post-operative menstrual loss and dysmenorrhoea, and the need for further surgery. Data on side-effects were also recorded. MAIN RESULTS: When compared with no treatment GnRH analogues are associated with a shorter duration of surgery, greater ease of surgery and a higher rate of post-operative amenorrhoea. Post-operative dysmenorrhoea also appears to be reduced. The use of GnRH analogues has no effect on intra-operative complication rates and patient satisfaction with this surgery is high irrespective of the use of any pre-operative endometrial thinning agent. GnRH analogues produce more consistent endometrial atrophy than danazol. For other intra-operative and post-operative outcomes any differences are minimal. Both GnRH analogues and danazol produce side-effects in a significant proportion of women, though few studies have reported these in detail. Little randomised data is available to assess the effectiveness of progestogens as endometrial thinning agents and the effect of any thinning agent

Danazol↗
PubMed2003

Videoradiographic analysis of how carbonated thin liquids and thickened liquids affect the physiology of swallowing in subjects with aspiration on thin liquids.

PURPOSE: To analyze how carbonated thin liquids affected the physiology of swallowing in dysphagic patients. MATERIAL AND METHODS: 40 patients were analyzed; 36 were neurologically impaired. During a therapeutic videoradiographic swallowing examination the patients had to swallow liquids with the following consistencies three times: thin, thickened and carbonated. The liquids were given in doses of 3 x 5 ml. The swallows were analyzed regarding penetration/aspiration, pharyngeal transit time and pharyngeal retention. RESULTS: Significant difference was found regarding penetration/aspiration when comparisons were made between thin liquid and carbonated thin liquid (p<0.0001). Carbonated liquid reduced the penetration to the airways. The comparison between thin liquid and thickened liquid (p<0.0001) showed significant less penetration with thickened liquids. Pharyngeal transit time was reduced both when comparing thin liquid with thin carbonated liquid (p<0.0001) and thickened liquid (p<0.0001). Pharyngeal retention was significantly reduced (p<0.0001) with carbonated thin liquid compared to thickened liquid. The comparison of thin liquids and carbonated thin liquids showed p=0.0013, thin and thickened liquids p=0.0097. CONCLUSIONS: Carbonated liquids reduced penetration/aspiration into the airways, reduced pharyngeal retention and pharyngeal transit time became shorter. Therefore, carbonated liquids are a valuable treatment option for patients with penetration/aspiration. Thickened liquids may still be an option for patients who cannot tolerate carbonated liquids and liquids with this consistency are safer than thin liquids.

Aged↗
PubMed1995

Mode of caldesmon binding to smooth muscle thin filament: possible projection of the amino-terminal of caldesmon from native thin filament.

The structure of smooth muscle thin filament was examined by various electron microscopy techniques, with special attention to the mode of caldesmon binding. Chemical cross-linking was positively used to avoid the dissociation of accessory proteins upon dilution. Caldesmon in reconstituted thin filament was observed as fine filamentous projections from thin filament. Native thin filament isolated from smooth muscle showed similarly numerous fine whisker-like projections by all the techniques employed here. Antibody against the amino-terminus of caldesmon labeled the end of such projections indicating the possibility that the amino-terminal myosin binding moiety might stick out from the shaft of the thin filament. Such whiskers are often projected out as a cluster to the same side of native thin filament. Further, we could visualize the assembly of dephosphorylated heavy meromyosin (HMM) with native or reconstituted thin filament forming "nonproductive" complex in the presence of ATP. The association of HMM to the shaft of thin filament was through subfragment-2 moiety, in accordance with biochemical studies. Some HMM particles bound closer to the thin filament shaft, possibly suggesting the presence of the second myosin-binding site on caldesmon. Occasionally two kinds of HMM association as such coexisted at a single site on this filament in tandem. Thus, we constructed a structural model of thin filament. The proposed molecular arrangement is not only compatible with all the biochemical results but also provides additional support for our recent findings (E. Katayoma, G. C. Scott-Woo, and M. Ikebe (1995) J. Biol. Chem. 270, 3919-3925) regarding the capability of caldesmon to induce dephosphorylated myosin filament, which explains the existence of thick filaments in relaxed smooth muscle cells.

Actin Cytoskeleton↗
PubMed1996

Consideration of a thin flap as an entity and clinical applications of the thin anterolateral thigh flap.

A defatted (thinned) anterolateral thigh flap was designed to reconstruct skin defects requiring thin flap coverage. We used this flap as a free flap for five cases of skin defects, and the outcomes of the reconstructions were all successful. The vascular pedicle of this flap, the cutaneous perforator of the lateral circumflex femoral artery, is about 8 cm long and 2 mm in diameter, and it is ideal for microvascular anastomosis. Thinning is performed in about 3 to 4 mm of thickness almost uniformly except for the vascular pedicle. It was ascertained as one of the useful donor sites of the free thin flap. The virtue of the thin anterolateral thigh flap is its uniform thinness compared with other thin flaps reported previously--the thin groin flap and the thin rectus abdominis musculocutaneous flap. We considered thin flaps as an entity, and they are classified into three types.

Adult↗
PubMed1991

Nebulin as a length regulator of thin filaments of vertebrate skeletal muscles: correlation of thin filament length, nebulin size, and epitope profile.

Nebulin, a family of giant proteins with size-variants from 600 to 900 kD in various skeletal muscles, have been proposed to constitute a set of inextensible filaments anchored at the Z line (Wang, K., and J. Wright. 1988. J. Cell Biol. 107:2199-2212). This newly discovered filament of the skeletal muscle sarcomere is an attractive candidate for a length-regulating template of thin filaments. To evaluate this hypothesis, we address the question of coextensiveness of nebulin and the thin filament by searching for a correlation between the size of nebulin variants and the length distribution of the thin filaments in several skeletal muscles. A positive linear correlation indeed exists for a group of six skeletal muscles that display narrow thin filament length distributions. To examine the molecular and architectural differences of nebulin size-variants, we carried out immunoelectron microscopic studies to map out epitope profiles of nebulin variants in these muscles. For this purpose, a panel of mAbs to distinct nebulin epitopes was produced against rabbit nebulin purified by an improved protocol. Epitope profiles of nebulin variants in three skeletal muscles revealed that (a) nebulin is inextensible since nebulin epitopes maintain a fixed distance to the Z line irrespective of the degree of sarcomere stretch; (b) a single nebulin polypeptide spans a minimal distance of 0.9 microns from the Z line; (c) nebulin contains repeating epitopes that are spaced at 40 nm or its multiples; (d) nebulin repeats coincide with thin filament periodicity; (e) nebulin variants differ mainly at either or both ends; and (f) nebulin remains in the sarcomere in actin-free sarcomeres produced by gelsolin treatment. Together, these data suggest that nebulin is an inextensible full-length molecular filament that is coextensive with thin filaments in skeletal muscles. We propose that nebulin acts as a length-regulating template that determines thin filament length by matching its large number of 40-nm repeating domains with an equal number of helical repeats of the actin filaments.

Animals↗
PubMed2002

Novel method for the plan-view TEM preparation of thin samples on brittle substrates by mechanical and ion beam thinning.

A new preparation method has been developed in order to avoid the breaking of brittle samples for plan-view TEM investigation during and after mechanical and ion beam thinning. The thinning procedure is carried out on a reduced size piece of the sample (about 1.6 x 0.8 mm(2) or about 1-1.6 mm diameter) that is embedded into a 3-mm-diameter Ti disk, which fits the sample holder of the TEM. The small sample size and the supporting metal disk assure the mechanical stability and minimize the possibility of breaking during and after the preparation: The Ti disk is placed on adhesive kapton tape, a cut piece of the sample is put into the slot of the disk, pressed onto the tape and embedded with glue. The tape keeps the parts in place and in the same plane, keeps the sample surface safe from the embedding glue and can be removed easily after the glue solidifies. Subsequently, the embedded sample is thinned from the rear by well-known mechanical and ion beam techniques until electron transparency. This simple solution lowers the risk of failed sample preparation remarkably and makes it possible to reduce the thickness of the sample to about 50 microm by mechanical thinning. As a result, dimpling becomes unnecessary and low angle ion milling gives a large transparent area for TEM. Its efficiency has been proved by successful preparation of numerous thin film samples on Si, sapphire, and glass substrates. The method is compatible with the widespread cross-sectional thinning procedures, and can be easily adopted by TEM laboratories.

Microscopy, Electron↗
PubMed1989

Caldesmon and the structure of smooth muscle thin filaments: immunolocalization of caldesmon on thin filaments.

Antibodies reacting with chicken gizzard caldesmon were used to determine the distribution of caldesmon on smooth muscle thin filaments. Antibodies developed against both the intact caldesmon molecule and a 40 kilodalton proteolytic fragment cause thin filaments to aggregate laterally. Aggregates produced with the latter antibody display regular periodic labelling with a repeat of approximately 38 nm, a distribution characteristic of proteins associated with tropomyosin on thin filaments. The stoichiometry of caldesmon on thin filaments has been critically reevaluated and alternative models of caldesmon distribution on thin filaments are proposed.

Animals↗
PubMed1990

Caldesmon and the structure of smooth muscle thin filaments: electron microscopy of isolated thin filaments.

Native and synthetic vertebrate smooth muscle thin filaments have been examined by electron microscopy in order to determine the arrangement of the regulatory protein caldesmon. In synthetic filaments of actin-caldesmon, long slender molecules were sometimes seen running along the thin filament, suggesting that caldesmon can associate with actin along its length, while at other times lateral projections were observed. In native filaments, containing actin, caldesmon and tropomyosin, we found no evidence for lateral projections extending from the filaments, suggesting that caldesmon does not act as a crosslinking protein in vivo. In contrast, elongated molecules were clearly seen following the long pitch actin helices. We suggest that these may represent an association of caldesmon and tropomyosin. Antibodies developed against an N-terminal fragment of caldesmon caused thin filaments to aggregate laterally into arrays displaying approximately 35-38 nm repeats; thin filament aggregates with this periodicity were obtained previously (Lehman et al., 1989) using antibodies to the C-terminal segment of caldesmon. These results suggest that both ends of caldesmon are closely associated with the shaft of the thin filament, supporting a model in which the elongated caldesmon molecule runs along the filament, possibly interacting with tropomyosin, following the long pitch actin helices.

Animals↗
PubMed2003

To thin or not to thin: the use of the anterolateral thigh flap in the reconstruction of intraoral defects.

The anterolateral thigh (ALT) flap has achieved popularity recently for free-flap reconstruction of intraoral defects following excision of squamous cell carcinoma. We have assessed the feasibility of the ALT flap as a free flap for oral lining and the potential use of the thinned ALT flap in a one-stage reconstruction. We used the ALT flap to reconstruct the oral cavity in 18 consecutive patients between December 2000 and December 2001 following intraoral resection of squamous cell carcinoma. Twelve patients underwent reconstruction using a standard ALT flap, four patients received a thinned ALT flap in a one-stage procedure, one patient received a standard ALT flap in combination with a fibula flap and one patient received a combination of a standard ALT flap and vascularised iliac bone. There were no complications in any of the 14 cases in which a standard ALT flap was used. Two of these flaps were thinned subsequently as secondary procedures. Of the four thinned ALT flaps, one flap failed completely and two flaps experienced partial necrosis. In all but one case the donor site was closed directly with minimal donor-site morbidity. The ALT flap is a versatile flap that can be used in combination with other flaps for more complex defects with minimal donor-site morbidity and is a useful alternative in the armamentarium of the head and neck surgeon. Thinning of the flap is best performed as a secondary procedure, should it be required.

Aged↗
PubMed1986

The diamond knife "semi": a substitute for glass or conventional diamond knives in the ultramicrotomy of thin and semi-thin sections.

The diamond knife "semi" for ultramicrotomes was originally designed by its manufacturer (DIATOME S.A.), for cutting semi-thin sections from 0.2 micron to 2.0 micron. Cutting tests of Epon-embedded material (nervous system, myelin sheat) with this knife have shown that the quality of semi-thin sections is equivalent or better than that obtained with a glass knife, and much time could be saved during the microtomy of serial sections. The quality of thin sections (0.07 micron to 0.12 micron) is excellent and comparable to that obtained with a conventional diamond knife. Furthermore, when adjacent sections are cut thin and semi-thin (for immunochemistry or high voltage microscopy), both are excellent in that they are of uniform thickness. In conclusion, this tool has advantages for both light microscopy and transmission electron microscopy.

Animals↗
PubMed2001

Integration of microfabricated needle-type glucose sensor devices with a novel thin-film Ag/AgCl electrode and plasma-polymerized thin film: mass production techniques.

We developed an integrated array of needle-type biosensors employing a novel process of fabrication, comprising conventional semiconductor fabrication and micromachining technology. Amperometric sensing electrodes with plasma-polymerized films and a thin-film Ag/AgCl reference electrode were directly integrated on a glass substrate with thin-film process, e.g., sputtering. An enzyme was immobilized on the electrode via the plasma-polymerized film, which was deposited directly on the substrate using a dry process. The novel thin-film Ag/AgCl reference electrode showed stable potentials in concentrated chloride solutions for a long period. The plasma-polymerized film is considered to play an important role as an interfacial design between the sensing electrode and the immobilized enzyme considering that the film is extremely thin, adheres well to the substrate (electrode) and has a highly cross-linked network structure and functional groups, such as amino groups. The results showed increments of the sensor signal, probably because the plasma-polymerized film allowed a large amount of enzyme to be immobilized. The greatest advantage is that the process can permit the mass production of high-quality biosensors at a low cost.

Biosensing Techniques↗
PubMed1997

Thin-section CT obtained at 10-mm increments versus limited three-level thin-section CT for idiopathic pulmonary fibrosis: correlation with pathologic scoring.

OBJECTIVE: The purpose of our study was to determine if three-level thin-section CT depicts idiopathic pulmonary fibrosis (IPF) pathology as accurately as CT obtained at 10-mm increments throughout the entire lungs. SUBJECTS AND METHODS: Thin-section (1.0- to 1.5-mm) images at 10-mm increments were obtained and scored prospectively in 25 consecutive patients with newly diagnosed IPF who were participating in a Special Center of Research grant for interstitial lung disease. Each patient's lobe was scored by four thoracic radiologists on a scale of 0-5 for both ground-glass attenuation and fibrosis. The radiologists used three images (limited CT) and also used the entire data set (complete CT). CT scores were compared with pathology scores from 67 open and thoracoscopic biopsies. Limited and complete scores were compared with each other (Pearson correlation coefficient). Interobserver variation in the CT scoring system was assessed using kappa values. RESULTS: CT fibrosis scores strongly correlated with pathology fibrosis scores for complete (r = .53, p = .0001) and limited (r = .50, p = .0001) CT. CT ground-glass scores correlated with the histologic inflammatory scores for each lobe on complete (r = .27, p = .03) and limited (r = .26, p = .03) CT. The desquamative subcomponent of the pathology inflammatory score had the highest correlation with the CT ground-glass scores (complete: r = .29, p = .01; limited: r = .33, p = .007). Good interobserver agreement existed for both the alveolar and fibrosis components of the CT scoring system (kappa values ranging from .51 to .83) for each lobe of the lung on limited and complete CT. CONCLUSION: Limited thin-section CT reveals the pathologic changes associated with IPF as well as CT obtained at 10-mm increments. An added advantage of limited thin-section CT is that it exposes patients to less radiation.

Adult↗
PubMed1991

The expression of actin mRNA in megakaryocytes demonstrated in paraffin sections, and epoxy resin semi-thin and thin sections by in situ hybridization.

Megakaryocytes of dog bone marrow were utilized as target cells for identifying actin mRNA expressing cells on semi-thin and thin sections. After in situ hybridization with radioisotope-labeled probes was performed on paraffin sections, gelatin capsules containing freshly prepared epoxy resin were placed on the sections. The resin was solidified and detached from the slide glass, and semi-thin and thin sections were obtained. The signals showing actin mRNA expression were detected on megakaryocytes in these sections by light and electron microscopy.

Actins↗
PubMed1988

Separation of lipids by new thin-layer chromatography and overpressured thin-layer chromatography methods.

New methods were developed for the separation of major lipid classes varying in polarity from cholesterol esters to lysophosphatidylcholine. The methods were used for the analysis of extracts obtained from human sera. The lipids were separated by overpressured thin-layer chromatography, classical thin-layer chromatography, and one-dimensional thin layer chromatography, using six different solvent systems for development. These techniques are also suitable to separate unsaturated and saturated cholesterol esters according to the number of carbon atoms and double bond numbers of their constituent fatty acids.

Cholesterol Esters↗