Effect of acylation on the interaction of the N-Terminal segment of pulmonary surfactant protein SP-C with phospholipid membranes.

SP-C, the smallest pulmonary surfactant protein, is required for the formation and stability of surface-active films at the air-liquid interface in the lung. The protein consists of a hydrophobic transmembrane alpha-helix and a cationic N-terminal segment containing palmitoylated cysteines. Recent evidence suggests that the N-terminal segment is of critical importance for SP-C function. In the present work, the role of palmitoylation in modulating the lipid-protein interactions of the N-terminal segment of SP-C has been studied by analyzing the effect of palmitoylated and non-palmitoylated synthetic peptides designed to mimic the N-terminal segment on the dynamic properties of phospholipid bilayers, recorded by spin-label electron spin resonance (ESR) spectroscopy. Both palmitoylated and non-palmitoylated peptides decrease the mobility of phosphatidylcholine (5-PCSL) and phosphatidylglycerol (5-PGSL) spin probes in dipalmitoylphosphatidylcholine (DPPC) or dipalmitoylphosphatidylglycerol (DPPG) bilayers. In zwitterionic DPPC membranes, both peptides have a greater effect at temperatures below than above the main gel-to-liquid-crystalline phase transition, the palmitoylated peptide inducing greater immobilisation of the lipid than does the non-palmitoylated form. In anionic DPPG membranes, both palmitoylated and non-palmitoylated peptides have similar immobilizing effects, probably dominated by electrostatic interactions. Both palmitoylated and non-palmitoylated peptides have effects comparable to whole native SP-C, as regards improving the gel phase solubility of phospholipid spin probes and increasing the polarity of the bilayer surface monitored by pK shifts of fatty acid spin probes. This indicates that a significant part of the perturbing properties of SP-C in phospholipid bilayers is mediated by interactions of the N-terminal segment. The effect of SP-C N-terminal peptides on the chain flexibility gradient of DPPC and DPPG bilayers is consistent with the existence of a peptide-promoted interdigitated phase at temperatures below the main gel-to-liquid-crystalline phase transition. The palmitoylated peptide, but not the non-palmitoylated version, is able to stably segregate interdigitated and non-interdigitated populations of phospholipids in DPPC bilayers. This feature suggests that the palmitoylated N-terminal segment stabilizes ordered domains such as those containing interdigitated lipids. We propose that palmitoylation may be important to promote and facilitate association of SP-C and SP-C-containing membranes with ordered lipid structures such as those potentially existing in highly compressed states of the interfacial surfactant film.

Theoretical model to determine the effects of geometrical factors on the resorption of calcium phosphate bone substitutes.

A theoretical approach was used to determine the effect of geometrical factors on the resorption rate of calcium phosphate bone substitutes that are either dense, microporous, and/or contain spherical macropores. Two cases were considered: (a) macroporous blocks that can be invaded by resorbing cells either directly because the structure is fully open-porous, or indirectly after some resorption of the macropores walls and/or interconnections. (b) Microporous or dense blocks/granules that cannot be invaded by resorbing cells, i.e. can only be resorbed from the outside to the inside, layer by layer. The theoretical approach was based on five assumptions: (i) the pores are spherical; (ii) the pores are ordered according to a face-centered cubic packing; (iii) the resorption is surface-controlled; (iv) the resorption is only possible if the surface can be accessed by blood vessels of 50 microm in diameter; and (v) the resorption time of a given amount of calcium phosphate is proportional to the net amount of material. Based on these assumptions, the calculations showed that the resorption time of a macroporous block could be minimized at a specific pore radius. This pore radius depended (i) on the size of the bone substitute and (ii) on the interpore distance. Typical radii were in the range of 100-400 microm. These values are similar to the numerous pore size optima mentioned in the scientific literature. For microporous or dense blocks/granules, the model suggested that a relatively small radius should be preferred. Such a radius leads to an optimum combination of a high surface area favorizing resorption and the presence of large intergranular gaps favorizing blood vessel ingrowth. In that case, the optimum of granule radius is around 100-200 microm. Finally, a very good agreement was found between the predictions of the model and experimental data, i.e. the model explained in all but two cases the results with an accuracy superior to 80%. In conclusion, the model appears to be a useful tool to better understand in vivo results, and possibly better define the geometry and distribution of the pores as well as the size of a bone substitute.

Technical innovations in medullary reaming: reamer design and intramedullary pressure increase.

BACKGROUND: Reaming the medullary cavity leads to an increase in intramedullary pressure, which can cause local necrosis and lung malfunction after intramedullary nailing. This investigation concentrates on the effect of reamer design parameters on intramedullary pressure increase. METHODS: Pressure measurements were obtained for three newly designed solid reamers and one hollow reamer. The AO/ASIF reamer was used for reference values. All reamers were connected with small flexible shafts. The pressures were measured in Plexiglas tubes filled with a mixture of Vaseline and paraffin oil with flow properties at 20 degrees C equivalent to those of bovine medullary fat at 36 degrees C. The reaming assemblies were inserted into the tubes using a materials testing machine at a constant speed. In addition, pressure measurements were made using five pairs of human femora. RESULTS: The highest pressures occurred with the AO/ASIF reamer head (258+/-29 mm Hg, p+/-0.05). By creating a conical form and enlarging the flutes, pressures were reduced by up to 37% (164+/-13 mm Hg, p < or = 0.05) compared with the AO/ASIF reamer, depending on the depth of the flutes. With a newly designed hollow reamer, pressure was reduced by 58% (108+/-19 mm Hg, p < or = 0.05) compared with the AO/ASIF reamer. CONCLUSION: The results show that optimizing the design of the reamer head leads to a significant reduction in pressure increase. These results should be taken into consideration when designing new reaming systems in an attempt to minimize the complication rate for intramedullary nailing.

Value of 31P NMR spectroscopy in predicting the response of a xenografted human hypopharynx carcinoma to irradiation.

PURPOSE: An early indicator of tumor sensitivity to irradiation could provide useful information on the effectiveness of therapy and may facilitate more individual designs of treatment protocols. The aim of the present study was to evaluate the potential of in vivo 31P nuclear magnetic resonance spectroscopy in predicting the response of a xenografted human hypopharynx carcinoma to radiotherapy. METHODS: The tumor had been serially heterotransplanted to athymic mice. 31P NMR spectra were collected before and at four intervals (24, 48, 72, and 120 h) after irradiation with 15 Gy or 30 Gy. Alterations of phosphorus metabolism were compared with the growth delays, the histological appearance, and the mitotic activity of the treated tumors. RESULTS: Radiation with 30 Gy induced increases of the phosphodiester level (P < 0.001) as well as of the tumor pH (P < 0.05) and decreases of the phosphomonoester level (P < 0.001) within 48 h. The changes clearly preceded measurable tumor responses and were accompanied by severe histological destruction and marked depression of mitotic indices. However, none of these spectral alterations was significantly correlated with individual delays of tumor growth. The only parameters allowing a prediction of radiation-induced tumor responses were the pre-treatment levels of phosphomonoesters and -diesters. The 31P NMR spectroscopic changes observed after therapy with 15 Gy were either unsystematic or insignificant. CONCLUSIONS: Pretreatment levels of tumor phospholipids were indicative of radiosensitivity in the xenografted human hypopharynx carcinoma investigated here. However, since phosphorus metabolism varies considerably among different tumor lines, it seems unlikely that there exists a uniform 31P NMR spectroscopic parameter predicting tumor response to radiation therapy.

[Biomechanics of interlocked nailing in humeral shaft fractures. Comparison of 2 nail systems and the effect of interfragmentary compression with the unreamed humeral nail]. / Biomechanik der Verriegelungsmarknagelung bei Oberarmschaftfrakturen. Vergleichsuntersuchungen zweier Marknagelsysteme und des Effekts der interfragmentären Kompression beim unaufgebohrten Humerusnagel.

In this biomechanical study the implanted Unreamed Humeral Nail (UHN) has been tested concerning bending and torsional stiffnesses. In literature other intramedullary implants have been criticized for insufficient rotatory stability especially in transverse and short oblique fractures of the humeral shaft. This study examined, whether the implanted UHN, as well as the UHN implanted with interfragmentary compression through a specific compression device, is able to augment torsional stiffness significantly. To evaluate bending and torsional stiffnesses, the UHN has been compared biomechanically to the Russell-Taylor humeral nail (RT) in paired mid-shaft osteotomized cadaveric humeri. Identic paired comparison has been performed with the UHN without and UHN with interfragmentary compression. In anterior-posterior, as well as medio-lateral direction stiffness under four-point-bending is significantly higher in stabilizing with the RT. Under torsional loading with moments of 4 Nm, 6 Nm and 8 Nm the UHN reached more than the double torsional stiffness. The RT, which is only dynamically interlocked, owns a high initial "play" between bolts and nail itself. Through additional interfragmentary compression stiffness of the UHN under four-point-bending in anterior-posterior, as well as medio-lateral direction augments significantly. Also under torsional loading with moments of 4 Nm, 6 Nm und 8 Nm torsional stiffness increases with interfragmentary compression significantly. In comparison to other biomechanical studies of different authorship it is clear, that this statically interlocked intramedullary nailing of the humeral shaft is superior to non-statically interlocked types of nailing concerning their stabilizing potency in torsion and serves especially for fracture types, which are critically under rotation, as transverse or short oblique humeral shaft fractures.

Functional dissociation between medial and lateral prefrontal cortical spatiotemporal activation in negative and positive emotions: a combined fMRI/MEG study.

The orbitofrontal cortex has been cytoarchitectonically and connectionally subdivided into a medial and a lateral part which are assumed to subserve distinct functions in emotional processing. However the exact spatiotemporal mechanisms of negative and positive emotional processing in medial and lateral orbitofrontal cortex remain unclear. We therefore investigated spatiotemporal orbitofrontal and prefrontal cortical activation patterns during emotional stimulation in a combined fMRI/MEG study. We investigated 10 healthy subjects, 5 women and 5 men. Positive and negative pictures from the International Affective Picture system (IAPS) were used for emotional stimulation, whereas neutral and gray pictures were taken as control conditions. fMRI/MEG measurements covered the whole frontal lobe and a time window between -2000 and +200 ms around motor responses (right index finger extension) associated with each picture. Positively and negatively correlated activities were determined in various prefrontal/frontal cortical regions in fMRI. Isocontour maps and single dipoles in MEG were analyzed in 50 ms time windows ranging from -2000 to +200 ms. Dipoles and fMR images were mapped on three-dimensional anatomical MRI so that anatomical localization of single dipoles and regional fMRI activity could be compared. Both negative and positive emotional conditions differed from non-emotional control conditions by strong orbitofrontal and lateral prefrontal activation as well as by the presence of early magnetic fields (-1700 to +1100 ms). Negative emotional processing was characterized by strong medial orbitofrontal activation and earlier (-1700 ms), stronger and more medially oriented orbitofrontal dipoles. In contrast positive emotional processing showed a rather strong activation in lateral prefrontal cortex with later (-1500 ms), weaker and more laterally oriented orbito and prefrontal dipoles. Negative emotional processing can be characterized by strong and early medial orbitofrontal cortical activation, whereas positive emotional processing showed rather later and weaker activation in lateral orbitofrontal/prefrontal cortex. Such a functional dissociation between medial and lateral orbito-frontal/prefrontal cortex during negative and positive emotional processing lends further support to the assumption of a functional subdivision in the orbitofrontal cortex.

Biomechanical comparison of bending and torsional properties in retrograde intramedullary nailing of humeral shaft fractures.

OBJECTIVE: To establish whether the bending and torsional stiffness of an implanted nail are influenced by nail design and nail-bolt interface, this study compared two implanted retrograde nail systems: the AO/ASIF unreamed humeral nail (UHN) and the Russell-Taylor (RT) nail. DESIGN: Pair randomization. SETTING: Mechanical laboratory testing. SPECIMENS: Twelve pairs of freshly harvested cadaveric humeri. METHODS: Transverse fractures were simulated with a standardized midshaft osteotomy and a three-millimeter gap. Both nails were proximally and distally interlocked. The RT nail has a single interlock at its base and tip. The UHN has double interlocking both proximally and distally. The screw hole design of the RT nail features slots, whereas the UHN has round screw holes. MAIN OUTCOME MEASURES: Anteroposterior and mediolateral bending stiffness and torsional stiffness. RESULTS: The RT nail showed higher bending stiffness in anteroposterior and mediolateral bending. Large differences were seen in the torsional characteristics: for the first 30 degrees, the RT nail showed a much lower resistance against torsion than the UHN. Analysis of variance of stiffness at four, six, and eight newton-meters showed statistical significance (p < 0.0001). Torsional stiffness, defined as the slope of a straight line approximated to between 75 and 100 percent of the maximum torque, was very similar in both nails. CONCLUSION: The torsional differences between the two nail systems are attributable to the nail-bolt interface of the RT nail. This dynamic system allows a clinically relevant degree of movement. The greater resistance to rotatory forces of the UHN is explained by the fact that the interlocking at its tip and base creates a static rather than a dynamic system.

Electrodynamic headphones and woofers for application in magnetic resonance imaging scanners.

Electrodynamic speakers compatible with (functional) magnetic resonance imaging (MRI) are described. The speakers magnets are removed, their function is replaced by the scanner's magnetic field, resulting in an uncommon but efficient operation. The method can be used with headphones as well as woofers. Functional MRI is not associated with any known biological risks, but as a method for visualization of task-specific activation of brain regions it is undesirably noisy. Thus, it requires both noise protection and efficient sound transmission systems for delivering acoustic stimuli to subjects. Woofers could possibly be used in active noise-control systems. The speakers described in this paper can be used for either task.

Functional magnetic resonance imaging of a human auditory cortex area involved in foreground-background decomposition.

Auditory foreground-background decomposition is a pattern recognition process which combines simultaneous and sequential grouping in complex sound sequences. Using functional magnetic resonance imaging with reduced scanner noise and stimulation through a new type of earphones, we investigated the possibility that this process activates topographically distinct areas of human auditory cortex. A basic matching-to-sample task with variable tones (sequential grouping) caused significant activity in three separate landmark-related territories on the supratemporal plane. A similar task in the presence of a strongly masking acoustic background pattern to challenge simultaneous grouping led to the distinction of the subterritory in which foreground signal-related or task-related signal properties were exclusively seen. In contrast to the remainder of territories the level of activity and the periodicity of the signal time-course was resistant to the masking influence of the background. This suggests that auditory foreground-background decomposition involves a specialized non-primary auditory cortex field. Generally, the findings demonstrate functional parcellation of auditory cortex for which the evidence in humans, in contrast to other primates, is only indirect to date.

Activation of human auditory cortex in retrieval experiments: an fMRI study.

In a previous functional magnetic resonance (fMRI) study, a subdivision of the human auditory cortex into four distinct territories was achieved. One territory (T1a) exhibited functional specialization in terms of a foreground-background decomposition task involving matching-to-sample monitoring on tone sequences. The present study more specifically determined whether memory-guided analysis of tone sequences is part of the T1a specialization. During the encoding periods, an arbitrary and unfamiliar four-tone-sequence (melody) played by one instrument was presented. The melody-instrument-combination was different in each period. During subsequent retrieval periods, learned and additional combinations were presented, and the tasks were either to detect the target melodies (experiment I) or the target instruments (experiment II). T1a showed larger activation during the melody retrieval. The results generally suggest that (1) activation of T1a during retrieval is determined less by the sound material than by the executed task, and (2) more specifically, that memory-guided sequential analysis in T1a is dominant over recognition of characteristic complex sounds.

Heat-induced segmental necrosis after reaming of one humeral and two tibial fractures with a narrow medullary canal.

In three cases referred to our clinic (a simple fracture of the humeral shaft, a simple, closed fracture, and a wedge fracture of the mid-third of the tibia), bone necrosis had resulted from excessive heat produced by reaming extremely narrow medullary cavities (5-5.5 mm diameter) with the 9 mm front-cutting reamer as part of a reamed nailing procedure. In any one case, different degrees of damage can occur from the metaphysis to the diaphysis. Based on the clinical course and the histological evaluation, we postulate that heat-induced damage can be divided into four degrees of severity (0-3): Grade 0: no damage; no devascularization, no heat-induced damage. Grade 1: The heat damaged zone is cut away during subsequent reaming, the only damage is devascularization. Grade 2: The damaged zones are not eliminated by subsequent reaming. The bone is devascularized and heat damaged. Grade 3: The entire cross section of the bone including the periosteum is devitalized by exposure to excessive heat. Depending on the severity of additional damage to the soft tissues, grave consequences are to be expected and further operations are unavoidable. The effects of heat-induced damage are particularly critical in the presence of infection (cases 2 and 3). The fundamental aspects and the extent of heat necrosis will be discussed. After discussion with the AO Technical Commission on the cause of heat-induced necrosis, we would recommend the following preventive measures: 1. preoperative measurement of the smallest diameter of the medullary cavity in two planes. 2. reaming with the standard instrumentation (9 mm) only if the medullary cavity has a diameter of at least 8 mm at its narrowest point. 3. Extremely narrow cavities should first be reamed manually or an alternative to nailing should be sought. 4. It is strongly recommended that only sharp reamers be used in such cases and blunt or damaged reamers replaced.

The physics of heat generation during reaming of the medullary cavity.

Clinical experience of special cases of reaming of a small diameter medullary cavity has shown a remarkable temperature increase with associated thermal damage to the surrounding tissue. By applying known solutions of the heat conductivity equation to simplified models of a reamer to bone tube system, the effects of the most important parameters on heat generation and temperature distribution are explained. The results allow a quantitative assessment of reaming techniques for small cavities which gives rise to the following recommendations: reamers used consecutively should not increase more than 0.5 mm in diameter; blunt instruments must be replaced or sharpened; prepare very narrow medullary cavities with the small diameter hand reamer; if the reamer ceases to advance do not increase the axial force but withdraw it and check the reamer and the temperature. Cooling, cleaning and replacement of the reamer should be considered, if appropriate.

Statistical methods in functional magnetic resonance imaging with respect to nonstationary time-series: auditory cortex activity.

In awake animal and human auditory cortices, it is a common experience with electrophysiological and suitable imaging methods for responses to steady stimulation to be strongly state-dependent and to exhibit nonstationarities, even over short periods of observation. If such nonstationary behavior is also reflected by hemodynamic responses in the human auditory cortex, conventional methods of analysis of fMRI data, although applicable for instance to largely stationary responses in visual and other cortices, may be misleading in attempts to parcellate auditory cortex into fields and to demonstrate functional maps. Time-Windows, described in this article as a convenient tool for the detection and analysis of time-variant brain activities, solves some of these problems. Time-Windows demonstrates that activity is evoked reliably in three separate territories of human auditory cortex, parts of which may show nonstationary behavior, depending on the auditory stimuli and tasks.

The hybrid ring tubular external fixator: a biomechanical study.

OBJECTIVE: To measure and compare the mechanical properties in bending of the four-ring, and three-ring/one-tube hybrid external fixation frames. DESIGN. IN VITRO: measurements of the mechanical behaviour of ring and ring-tubular external fixation frames. In the latter, one ring of the full circular frame was replaced by one tube and Schanz screws. BACKGROUND: The mechanical properties of the classical Ilizarov four-ring external fixation frames has been compared to those of other external fixation frames by various authors. However, in clinical practice the hybrid fixation frame is being used with increasing frequency. Therefore the mechanical properties of the latter are of immediate interest and clinical value. METHODS: On explanted sheep tibiae with single and double osteotomies, frame stiffness in the four-point bending mode was measured at different K-wire tensions, comparing the values obtained from four-ring frames, to those of three-ring-tubular hybrid frames. These measurements were made under conditions of (a) bone distraction (BD), and (b) segment transport (ST), both at the initial and final stages of this procedure. RESULTS: In circular frames, frame stiffness in bending for increasing K-wire tension showed a Gaussian distribution both in distraction and post-ST with an optimum at 1000 N. In ring tubular hybrid frames, however, frame stiffness showed a more linear relationship to K-wire tension. CONCLUSIONS: In the four-ring Ilizarov external fixation frame, the exchange of one ring with one tube and one Schanz screw both reduced frame stiffness in bending and converted to linear its relationship to K-wire tension. RELEVANCE: Under clinical conditions, the use of a similar ring tubular hybrid external fixator allows the adjustment of frame stiffness in a simple and practical way. This is not the case with the original ring fixation frame.

Lateralized processing of speech prosodies in the temporal cortex: a 3-T functional magnetic resonance imaging study.

Prosodic modulation of speech provides information about emotional states of speakers (affective prosodies) or serves as syntactic markers to change linguistic aspects of speech (linguistic prosodies). Previous electrophysiological investigations and studies on patients with right or left hemisphere damage showed nonuniform results with respect to lateralization of prosodic processing. In this study 20 healthy right-handed volunteers were investigated with functional magnetic resonance imaging of the acoustically responsive areas on the supratemporal plane while detecting phonemes as control targets or prosodies in strings of nonsense syllables and adjectives, the latter randomly intonated in a declarative, interrogative, commanding, happy, or sad fashion. In control task A the phoneme /a/ was detected in the syllables. In control task B the phoneme /a/ was detected in the adjectives, and in the experimental task C the sad intonations (affective) and in the experimental task D the interrogative intonations (linguistic) had to be detected in the same material. In task A intensity-weighted volumes of activated voxels were not different in the two hemispheres (laterality index 0). In task B with an irrelevant phoneme detection with respect to prosodic material, the population split into two subgroups with similar right or left hemispheric lateralization of activity leading to an absolute laterality index of 26.8 across all subjects. During detection of affective prosodies (task C), lateralization was maintained yet the absolute laterality index reduced to 14.5, while there was no lateralization during detection of linguistic prosodies. The sum of activations in the two hemispheres was the same across all tasks and subgroups, which suggests that the lateralizations occurring with presentation and detection of prosodic material depend on a redistribution of activity between hemispheres.

5-Fluorouracil metabolism and cytotoxicity after pre-treatment with methotrexate or thymidine in human hypopharynx and colon carcinoma xenografts: a 19F-nuclear magnetic resonance spectroscopy study in vivo.

The metabolism of 5-fluorouracil (5-FU) was monitored non-invasively in two xenografts, a hypopharynx carcinoma and a colon carcinoma (CSM) by 19F-magnetic resonance spectroscopy following an i.v. bolus injection of 130 mg kg-1 5-FU. Both the level of fluoronucleotides (FNuc) and the tumor growth delay were significantly higher in the CSM colon carcinoma than in the hypopharynx carcinoma (both parameters, P < 0.001). Administration of 100 mg kg-1 methotrexate (MTX) at 15 h before treatment with 5-FU caused a significantly increased conversion of 5-FU to FNuc in both tumors (P < 0.05) as compared with the application of 5-FU alone. However, only in the CSM tumor was a significantly increased growth delay (P < 0.01) observed. Pre-treatment of both xenografts with 400 mg kg-1 thymidine enhanced the conversion of 5-FU to FNuc in both tumors. In the CSM tumour this treatment modality caused a significantly (P < 0.05) higher growth delay as compared with the results obtained with 5-FU alone, whereas in the hypopharynx carcinoma the additional application of thymidine caused no significant change in tumor growth. It is known that both thymidine and MTX can reduce the DNA-directed cytotoxicity of 5-FU, whereas the RNA-directed cytotoxicity is increased. It is concluded that the DNA-mediated toxicity may be more important in the hypopharynx carcinoma than in the CSM colon carcinoma. As a consequence, pre-treatment with MTX or thymidine enhances FNuc formation, although only in the CSM carcinoma is there an increased tumor growth delay. Thus, in the hypopharynx carcinoma the measurement of FNuc did not serve as a predictor for the treatment efficacy of the combined treatment modality. Pre-treatment with MTX did not influence the catabolism of 5-FU, whereas thymidine actually prolonged the half-life of 5-FU without alpha-fluoro-beta-alanine becoming detectable.

A retrospective analysis of plate contouring in the tibia using the conventional 4.5 (narrow) dynamic compression plate.

Plate contouring is an essential part of plate osteosynthesis. Its safety and technical ease is dependent on plate factors (material and design) and the complexity of the contouring. Our purpose was to determine the range of plate contouring; estimate the strain on the plate from the radius of curvature in the area of contouring; and determine whether shaped plates had a reduction in their load carrying capacity. A retrospective analysis of 500 radiographed tibias that had undergone osteosynthesis with the conventional dynamic compression plate was conducted to evaluate the radius of curvature and degree of bend in the plates, and to determine whether plates were bent at screw holes. Contouring of the plate to fit the bone was done by the surgeon at the time of application. Postoperative radiographs were evaluated. We found that (a) the shorter the length of plate contoured, the greater the strain; (b) plates were bent at screw holes, and these tended to be the areas of highest strain; (c) the range of plate strain was 0.6-16% (average 1.6%); and (d) plates were bent from 10 to 90 degrees (average 20 degrees).