Validity and reliability of a portable handheld dynamometer compared to a fixed isokinetic dynamometer to assess forearm torque strength.

To evaluate the effect of treatment on forearm rotation, torque muscle strength can be assessed using an isokinetic device (IKD) or a wrist dynamometer (WD). The aims of this study were 1) to determine concurrent validity and intra- and inter-rater reliability using the WD, and to examine correlations between WD and IKD in different positions; and 2) subsequently, to establish the intermethod reproducibility between WD as a handheld (HHD) or fixed device. We conducted a cross-sectional study in which torque strength was measured in healthy participants by two observers using an IKD and a WD. Study endpoints were concurrent validity (Pearson's r), intra- and inter-rater reliability, intermethod reproducibility (intraclass correlation coefficient: ICC) and measurement error (limits of agreement: LoA). Concurrent validity ranged, in the 2 studies assessing it, from r 0.37 to 0.52 for pronation and from r 0.50 to 0.82 for supination, with wide 95% confidence intervals. ICC for intra-rater reliability for pronation ranged from 0.85 to 0.91 and for supination from 0.91 to 0.95. ICC for inter-rater reliability for pronation ranged from 0.84 to 0.96 and for supination from 0.92 to 0.96. Despite the excellent intra- and inter-rater reliability and intermethod reproducibility for the WD-HHD and fixed WD, validity was low when compared to IKD and wide LoA indicated a high measurement error of approximately 20%. These results suggest that the WD cannot replace the IKD isometric mode for pronation and supination. LEVEL OF EVIDENCE: 2.

Compressive strength of two newly developed glass-ionomer materials for use with the Atraumatic Restorative Treatment (ART) approach in class II cavities.

OBJECTIVES: The null-hypotheses tested were that no difference in compressive strength of ART class II cavities exists between those restored with (1) glass-carbomer and a commonly used glass-ionomer; (2) KMEM and the commonly used glass-ionomer and; (3) glass-carbomer and KMEM. METHODS: 100 molar teeth, stratified by size, were randomly allocated to the four test groups. Large ART class II cavities were drilled and restored with Clearfil photoposterior (negative control), Fuji IX (positive control), Glass-carbomer and Ketac Molar Easymix (KMEM) (experimental groups). Half of the samples in each test group were 5000 times thermocycled between 5 degrees C and 55 degrees C, with a 30s dwell time in each bath and a transfer time of 10s. The restorations were statically tested at the marginal ridge until failure, using a rounded rectangular testing rod at crosshead speed of 1.0mm/min. ANOVA and Student's t-test were applied to test for differences between the dependent variable (compressive strength at the final breaking point) and the independent variables (thermocycling and restorative material). RESULTS: Restorations of Clearfil photoposterior had a statistically significant higher mean compressive strength value at final breaking point than those of the three glass-ionomers tested (p=0.0001). No thermocycling effect was observed (p=0.19). ANOVA between the three glass-ionomer materials and mean compressive strength at final breaking point showed no statistically significant difference (p=0.09). SIGNIFICANCE: Class II ART cavities restored with the newly launched Glass-carbomer and Ketac Molar Easymix were not significantly more fracture resistant than comparable restorations using the conventional glass-ionomer Fuji IX.

Plasmodium vivax CS peptides display conformational preferences for folded forms in solution.

Spectroscopic techniques have been used to study the conformations of several synthetic peptides with sequences corresponding to the repeat regions of the circumsporozoite proteins of Plasmodium vivax, variants vk-210 and vk-247. As has previously been shown for P. falciparum, turn-like folded conformations are observed, in rapid dynamic equilibrium with extended-chain forms. These results are consistent with the known similarity of the structural, biosynthetic and immunological properties of the circumsporozoite proteins of different plasmodial species. Additionally, the observation of folded conformers provides a rationale for the effectiveness of these peptides as immunogens and potential vaccines.

Potential bias in NMR relaxation data introduced by peak intensity analysis and curve fitting methods.

We present an evaluation of the accuracy and precision of relaxation rates calculated using a variety of methods, applied to data sets obtained for several very different protein systems. We show that common methods of data evaluation, such as the determination of peak heights and peak volumes, may be subject to bias, giving incorrect values for quantities such as R1 and R2. For example, one common method of peak-height determination, using a search routine to obtain the peak-height maximum in successive spectra, may be a source of significant systematic error in the relaxation rate. The alternative use of peak volumes or of a fixed coordinate position for the peak height in successive spectra gives more accurate results, particularly in cases where the signal/noise is low, but these methods have inherent problems of their own. For example, volumes are difficult to quantitate for overlapped peaks. We show that with any method of sampling the peak intensity, the choice of a 2- or 3-parameter equation to fit the exponential relaxation decay curves can dramatically affect both the accuracy and precision of the calculated relaxation rates. In general, a 2-parameter fit of relaxation decay curves is preferable. However, for very low intensity peaks a 3 parameter fit may be more appropriate.

Sequence-dependent correction of random coil NMR chemical shifts.

Random coil chemical shifts are commonly used to detect secondary structure elements in proteins in chemical shift index calculations. While this technique is very reliable for folded proteins, application to unfolded proteins reveals significant deviations from measured random coil shifts for certain nuclei. While some of these deviations can be ascribed to residual structure in the unfolded protein, others are clearly caused by local sequence effects. In particular, the amide nitrogen, amide proton, and carbonyl carbon chemical shifts are highly sensitive to the local amino acid sequence. We present a detailed, quantitative analysis of the effect of the 20 naturally occurring amino acids on the random coil shifts of (15)N(H), (1)H(N), and (13)CO resonances of neighboring residues, utilizing complete resonance assignments for a set of five-residue peptides Ac-G-G-X-G-G-NH(2). The work includes a validation of the concepts used to derive sequence-dependent correction factors for random coil chemical shifts, and a comprehensive tabulation of sequence-dependent correction factors that can be applied for amino acids up to two residues from a given position. This new set of correction factors will have important applications to folded proteins as well as to short, unstructured peptides and unfolded proteins.

Local structural plasticity of the prion protein. Analysis of NMR relaxation dynamics.

A template-assisted conformational change of the cellular prion protein (PrP(C)) from a predominantly helical structure to an amyloid-type structure with a higher proportion of beta-sheet is thought to be the causative factor in prion diseases. Since flexibility of the polypeptide is likely to contribute to the ability of PrP(C) to undergo the conformational change that leads to the infective state, we have undertaken a comprehensive examination of the dynamics of two recombinant Syrian hamster PrP fragments, PrP(29-231) and PrP(90-231), using (15)N NMR relaxation measurements. The molecular motions of these PrP fragments have been studied in solution using (15)N longitudinal (T(1)) and transverse relaxation (T(2)) measurements as well as [(1)H]-(15)N nuclear Overhauser effects (NOE). These data have been analyzed using both reduced spectral density mapping and the Lipari-Szabo model free formalism. The relaxation properties of the common regions of PrP(29-231) and PrP(90-231) are very similar; both have a relatively inflexible globular domain (residues 128-227) with a highly flexible and largely unstructured N-terminal domain. Residues 29-89 of PrP(29-231), which include the copper-binding octarepeat sequences, are also highly flexible. Analysis of the spectral densities at each residue indicates that even within the structured core of PrP(C), a markedly diverse range of motions is observed, consistent with the inherent plasticity of the protein. The central portions of helices B and C form a relatively rigid core, which is stabilized by the presence of an interhelix disulfide bond. Of the remainder of the globular domain, the parts that are not in direct contact with the rigid region, including helix A, are more flexible. Most significantly, slow conformational fluctuations on a millisecond to microsecond time scale are observed for the small beta-sheet. These results are consistent with the hypothesis that the infectious, scrapie form of the protein PrP(Sc) could contain a helical core consisting of helices B and C, similar in structure to the cellular form PrP(C). Our results indicate that residues 90-140, which are required for prion infectivity, are relatively flexible in PrP(C), consistent with a lowered thermodynamic barrier to a template-assisted conformational change to the infectious beta-sheet-rich scrapie isoform.

Random coil chemical shifts in acidic 8 M urea: implementation of random coil shift data in NMRView.

Studies of proteins unfolded in acid or chemical denaturant can help in unraveling events during the earliest phases of protein folding. In order for meaningful comparisons to be made of residual structure in unfolded states, it is necessary to use random coil chemical shifts that are valid for the experimental system under study. We present a set of random coil chemical shifts obtained for model peptides under experimental conditions used in studies of denatured proteins. This new set, together with previously published data sets, has been incorporated into a software interface for NMRView, allowing selection of the random coil data set that fits the experimental conditions best.

Treatment of patients with long-standing unexplained subfertility with in vitro fertilization.

OBJECTIVE: To evaluate whether IVF is an effective treatment for long-standing unexplained subfertility. DESIGN: Retrospective cohort study. SETTING: Tertiary care infertility center in a university hospital. PATIENT(S): Two hundred two couples with unexplained subfertility of 2 years' duration or more who attended the center for their first IVF attempt. INTERVENTION(S): Couples were placed on a waiting list for IVF. They received no treatment until IVF was started. MAIN OUTCOME MEASURE(S): Pregnancy rate (PR) while on the waiting list and PR after IVF treatment. RESULT(S): Complete data sets were available for 131 couples. Seventeen of 131 women became pregnant while waiting for IVF treatment (PR 0.9% per exposure cycle), whereas 45 of 119 receiving IVF treatment became pregnant (PR 17% per IVF attempt). CONCLUSION(S): IVF treatment has substantial added value over waiting and is an efficient treatment for long-standing unexplained subfertility.

Mandibular movements in response to electrical stimulation of superficial and deep parts of the human masseter muscle at different jaw positions.

Anatomical and electromyographical evidence suggests a compartmentalized function of the human jaw-closing muscles during both static and dynamic motor tasks. However, the voluntary nature of these tasks hampers unequivocal interpretation of this evidence, because it is impossible to activate voluntarily a single part of a muscle exclusively. Activation of discrete, localized regions can be accomplished with electrical stimulation. A previous study confirmed a functional subdivision of the temporalis muscle into at least three parts. Here, differences in the direction of the lower incisal-point (IP) movement in response to electrical stimulation of four different parts of the masseter muscle were examined in five healthy men. The deep masseter muscle and the anterior, middle, and posterior parts of the superficial masseter muscle were stimulated with monopolar wire electrodes in four different jaw positions (resting position; 50% maximum mouth opening; and 10-mm right and left lateral excursions, both with respect to resting position). Electrode-insertion depth was measured from magnetic resonance images. Movement responses to stimulation were recorded with the OKAS-3D jaw-movement analysis system. The variation in the direction of the IP movement in response to stimulation of parts of the masseter was partly explained by the effects of stimulus location and jaw position. The response to stimulation of the deep masseter was mainly laterovertically directed, whereas the response to stimulation of each of the superficial parts had a mainly anterovertical direction, the responses being most pronounced with the mandible in its resting position. These results provide further evidence for a functional subdivision of the masseter into a superficial part and a deep part, but not for a further subdivision of the superficial part into an anterior, middle, and posterior part.

Randomised controlled trial of magnetic-resonance pelvimetry in breech presentation at term.

BACKGROUND: Pelvimetry is widely used in women with breech presentation at term to select those for whom planned vaginal delivery is appropriate. However, its clinical value has never been established. We evaluated pelvimetry in a randomised controlled trial. The main outcome measures were the elective and emergency caesarean-section rates and the early condition of the neonate. METHODS: Magnetic-resonance (MR) pelvimetry was done on 235 women. The women were then randomly assigned to two groups--for the study group (n = 118), the pelvimetry results were reported to the responsible obstetricians, who used them as the basis for decisions on whether to schedule elective caesarean or trial of labour; for the control group (n = 117), the pelvimetry results were not disclosed until 8 weeks post partum, and decisions about obstetric management were made on the basis of clinical factors only. FINDINGS: 35 women (15 [13%] study group, 20 [17%] control group) had abnormalities on pelvimetry. The overall caesarean-section rates did not differ significantly between the study and control groups (50 [42%] vs 59 [50%], p = 0.24) but the emergency caesarean-section rate was significantly lower in the study group than in the control group (22 [19%] vs 41 [35%], p = 0.0052). The mean 1 min Apgar scores in the study and control groups were 8.1 and 8.0 (p = 0.93) and the mean 3 min scores 9.5 and 9.4, respectively (p = 0.28). There were no significant differences in the early neonatal outcome for infants born vaginally, by emergency caesarean section, or by elective caesarean section in the two groups, except for a significantly lower Apgar score in the six infants born vaginally to control-group women who had pelvic abnormalities. INTERPRETATION: The use of MR pelvimetry in breech presentation at term did not significantly reduce the overall caesarean-section rate. However, it allowed better selection of the delivery route, with a significantly lower emergency caesarean-section rate. Neonatal outcome was not compromised by use of the pelvimetry data.

Jaw movement responses to electrical stimulation of different parts of the human temporalis muscle.

Previous EMG studies have provided indications for the differential activation of the human temporalis muscle. However, in these studies the contribution of different parts of the temporalis muscle could not be separated from the contributions of other muscles, since contraction was performed by voluntary activation. Therefore, the aim of this study was to examine functional differences among various parts of the human temporalis muscle by means of recording the incisal point movement response to electrical stimulation of parts of the muscle. Five healthy male subjects participated in this study. Three locations (anterior, middle, and posterior temporalis muscle) were stimulated, by means of monopolar wire electrodes and rectangular pulses. The insertion depths of the stimulation electrodes were determined by means of magnetic resonance images. Stimulation was performed in four jaw positions (resting position, 50% maximum mouth opening, 1 cm to the left, and 1 cm to the right). Movement responses to stimulation of the different muscle parts were recorded with the OKAS-3D jaw movement analysis system. The movement responses were expressed in polar coordinates. The variation in the direction of the jaw movement response was partly explained by the factors 'stimulation location' and 'jaw position' (ANOVA, p < 0.001). When the stimulation location shifted in an antero-posterior direction, the response changed from a vertical-lateral incisal point movement to a lateral-posterior movement with a smaller vertical component. The jaw position during stimulation also influenced the movement response. A functional subdivision of the temporalis muscle into at least three parts is favored.

EMG differences between weak and strong myogenous CMD patients and healthy controls.

Electromyographic and clinical characteristics of 42 myogenous craniomandibular disorder (CMD) patients were related to 40 healthy control subjects. Surface EMG recordings were obtained from the masseter and the temporal muscles. The clinical examination included active maximum mouth opening, endfeel distance, active laterotrusion and lateral endfeel distance, dental abrasion, the presence of a lateral slide (RCP-ICP) and the amount of static pain. Orthopantomographic X-rays were available from 32 CMD patients for measuring the condylar and ramus asymmetry. Compared to the control group lower masseter and temporal EMG amplitudes were found for myogenous CMD patients (P < 0.001). When the activity of the temporal muscle was compared with the activity of the masseter muscle, the CMD patients showed proportionally higher temporal muscle activities than the controls (P < 0.05, 50% clenching level). CMD patients also showed smaller mandibular excursions, larger endfeel distances and more dental abrasion than controls. The temporal muscle asymmetries showed significant negative correlations with the ramus asymmetries. The lower and upper quartile of the distribution of the mean masseter EMG amplitudes were used to distinguish weak and strong muscles in patients and controls. Weak patients showed proportionally high temporal muscle activities, larger masseter and temporal muscle asymmetries, and larger endfeel distances compared to strong CMD patients. These differences were not found between weak and strong controls. In conclusion, it can be said that the electromyographic and clinical findings of the myogenous CMD patients suggest a functional difference between weak and strong patients and indicate the need for more individually designed treatment modalities for functional muscle and joint CMD problems.

Modified assay of prostate-specific antigen with a detection limit < 0.01 microgram/L.

We modified the Hybritech Tandem-E prostate-specific antigen (PSA) assay by increasing the sample volume, increasing enzyme-substrate incubation time, and using diethanolamine buffer. Our modified method has a detection limit of 0.009 microgram/L (P < 0.01). The assay curve is linear from 0.01 to 1.0 micrograms/L and has an overall assay time of about 4 h. Linear plots are obtained when the 1.0 micrograms/L standard is diluted with either matrix buffer or serum from men containing PSA < 0.01 microgram/L. Recovery of PSA (0.10 microgram/L) added to serum from men averaged 94%. Interassay CVs were 13%, 7%, and 4% at PSA concentrations of 0.04, 0.07, and 0.30 micrograms/L, respectively (n = 33). This assay should be useful in the detection of early recurrence of prostate cancer after radical prostatectomy.

Backbone assignments and secondary structure of the Escherichia coli enzyme-II mannitol A domain determined by heteronuclear three-dimensional NMR spectroscopy.

This report presents the backbone assignments and the secondary structure determination of the A domain of the Escherichia coli mannitol transport protein, enzyme-IImtl. The backbone resonances were partially assigned using three-dimensional heteronuclear 1H NOE 1H-15N single-quantum coherence (15N NOESY-HSQC) spectroscopy and three-dimensional heteronuclear 1H total correlation 1H-15N single-quantum coherence (15N TOCSY-HSQC) spectroscopy on uniformly 15N enriched protein. Triple-resonance experiments on uniformly 15N/13C enriched protein were necessary to complete the backbone assignments, due to overlapping 1H and 15N frequencies. Data obtained from three-dimensional 1H-15N-13C alpha correlation experiments (HNCA and HN(CO)CA), a three-dimensional 1H-15N-13CO correlation experiment (HNCO), and a three-dimensional 1H alpha-13C alpha-13CO correlation experiment (COCAH) were combined using SNARF software, and yielded the assignments of virtually all observed backbone resonances. Determination of the secondary structure of IIAmtl is based upon NOE information from the 15N NOESY-HSQC and the 1H alpha and 13C alpha secondary chemical shifts. The resulting secondary structure is considerably different from that reported for IIAglc of E. coli and Bacillus subtilis determined by NMR and X-ray.

The NMR determination of the IIA(mtl) binding site on HPr of the Escherichia coli phosphoenol pyruvate-dependent phosphotransferase system.

The region of the surface of the histidine-containing protein (HPr) which interacts with the A domain of the mannitol-specific Enzyme II (II(Amt1)) has been mapped by titrating the A-domain into a solution of 15N-labeled HPr and monitoring the effects on the amide proton and nitrogen chemical shifts via heteronuclear single quantum correlation spectroscopy (HSQC). Fourteen of the eighty-five HPr amino acid residues show large changes in either the 15N or 1H chemical shifts or both as a result of the presence of II(Amt1) while a further seventeen residues experience lesser shifts. Most of the residues involved are surface residues accounting for approximately 25% of the surface of HPr. Phosphorylation of HPr with catalytic amounts of Enzyme I (EI), in the absence of II(Amt1) resulted in chemical shift changes in a sub-set of the above residues; these were located more in the vicinity of the active site phospho-histidine. Phosphorylation of the HPr/II(Amt1) complex resulted in a HSQC spectrum which was indistinguishable from the P-HPr spectrum in the absence of II(Amt1) indicating that, as expected, the complex P-HPr/P-II(Amt1) does not exist even at the high concentrations necessary for NMR.

Electromyographic evidence of local muscle fatigue in a subgroup of patients with myogenous craniomandibular disorders.

Ten female patients with myogenous craniomandibular disorders who had complete dentitions and mainly unilateral muscle pain and tenderness to palpation, and 10 healthy female controls, participated in the study. The surface electromyographic signals of masseter and anterior temporal muscles were recorded during 30-s test contractions at 50% of the maximum voluntary clenching effort. A force transducer placed between the central incisors recorded the maximum voluntary bite force. The patients exerted a lower maximum voluntary bite force than the controls (p less than 0.05). In the patients the electromyographic signals of the painful muscles were weaker than those of the painless muscles (p less than 0.01). The signals of the control subjects were stronger than those of the painless muscles of the patients (p less than 0.01). The mean power frequency of the signals, recorded at the start of the 30-s contractions, showed no differences between the painful and painless muscles and between the muscles of the patients and of the controls. The rate of mean power frequency shift in the electromyographic signal, as a response to the 30-s test contraction, was normalized with respect to the amplitude of that signal to account for its amplitude dependency. The painful muscles showed a higher normalized rate of mean power frequency shift than the painless muscles and the muscles of the control subjects (p less than 0.01). These results confirm the presence of an impaired condition in the painful muscles of this group of patients with myogenous craniomandibular disorders.

Conformation and mobility of the RNA-binding N-terminal part of the intact coat protein of cowpea chlorotic mottle virus. A two-dimensional proton nuclear magnetic resonance study.

Two-dimensional proton nuclear magnetic resonance (n.m.r.) experiments were performed on the coat protein of cowpea chlorotic mottle virus (molecular mass: 20.2 kDa) present as dimer (pH 7.5) or as capsid consisting of 180 protein monomers (pH 5.0). The spectra of both dimers and capsids showed resonances originating from the flexible N-terminal region of the protein. The complete resonance assignment of a synthetic pentacosapeptide representing this N terminus made it possible to interpret the spectra in detail. The capsid spectrum showed backbone amide proton resonances arising from the first eight residues having a flexible random coil conformation, and side-chain resonances arising from the first 25 N-terminal amino acids. The dimer spectrum showed also side-chain resonances of residues 26 to 33, which are flexible in the dimer but immobilized in the capsid. The n.m.r. experiments indicated that the conformation of the first 25 amino acids of the protein in dimers and capsids is comparable to the conformation of the synthetic peptide, which alternates among extended and helical conformations on the n.m.r. time-scale. It is suggested that the alpha-helical region, situated in the region between residues 10 and 20, binds to the RNA during assembly of the virus particle.

Recovery of the human biceps electromyogram after heavy eccentric, concentric or isometric exercise.

Five men performed submaximal isometric, concentric or eccentric contractions until exhaustion with the left arm elbow flexors at respectively 50%, 40% and 40% of the prefatigued maximal voluntary contraction force (MVC). Subsequently, and at regular intervals, the surface electromyogram (EMG) during 30-s isometric test contractions at 40% of the prefatigued MVC and the muscle performance parameters (MVC and the endurance time of an isometric endurance test at 40% prefatigued MVC) were recorded. Large differences in the surface EMG response were found after isometric or concentric exercise on the one hand and eccentric exercise on the other. Eccentric exercise evoked in two of the three EMG parameters [the EMG amplitude (root mean square) and the rate of shift of the EMG mean power frequency (MPF)] the greatest (P less than 0.001) and longest lasting (up to 7 days) response. The EMG response after isometric or concentric exercise was smaller and of shorter duration (1-2 days). The third EMG parameter, the initial MPF, had already returned to its prefatigued value at the time of the first measurement, 0.75 h after exercise. The responses of EMG amplitude and of rate of MPF shift were similar to the responses observed in the muscle performance parameters (MVC and the endurance time). Complaints of muscle soreness were most frequent and severe after the eccentric contractions. Thus, eccentric exercise evoked the greatest and longest lasting response both in the surface EMG signal and in the muscle performance parameters.