Interaction of actin and ADP with the head domain of smooth muscle myosin: implications for strain-dependent ADP release in smooth muscle

Transient kinetic methods were used to study interactions between actin, MgADP, and smooth muscle (chicken gizzard) myosin subfragment 1 (smS1). The equilibrium dissociation constant (Kd) of actin for smS1 was 3.5 nM, tighter than that of skeletal S1 (skS1). Actin binding to smS1 was weakened 5-fold by saturation with ADP compared to 30-60-fold for skS1. The Kd of ADP for smS1 was increased from 1.2 to 5 microM by actin, whereas for skS1 values increased from 2 to 100 microM. Thus, coupling between ADP and actin binding is weaker for smS1. Previous studies show that release of ADP from actin.smS1.ADP produces a tilt of the regulatory domain [Whittaker, M., Wilson-Kubalek, E. M., Smith, J. E., Faust, L., Milligan, R. A., and Sweeney, H. L. (1995) Nature 378, 748-751]. This result was confirmed by independent structural methods; tilting was absent for skS1, and the Kd for ADP was in agreement with the values measured here [Gollub, J., Cremo, C. R., and Cooke, R. (1996) Nat. Struct. Biol. 3, 796-802; Poole, K. I. V., Lorenz, M., Ellison, P., Evans, G., Rosenbaum, G., Boesecke, P., Holmes, K. C., and Cremo, C. R. (1997) J. Muscle Res. Cell Motility 18, 264]. We discuss tilting upon ADP release with respect to our measurements, previous measurements with skS1, and nucleotide concentrations in smooth muscle. We propose that these data suggest a strain-dependent ADP release mechanism that may be accentuated in smooth muscles.     

Genetic definition of a protein-splicing domain: functional mini-inteins support structure predictions and a model for intein evolution

Inteins are protein-splicing elements, most of which contain conserved sequence blocks that define a family of homing endonucleases. Like group I introns that encode such endonucleases, inteins are mobile genetic elements. Recent crystallography and computer modeling studies suggest that inteins consist of two structural domains that correspond to the endonuclease and the protein-splicing elements. To determine whether the bipartite structure of inteins is mirrored by the functional independence of the protein-splicing domain, the entire endonuclease component was deleted from the Mycobacterium tuberculosis recA intein. Guided by computer modeling studies, and taking advantage of genetic systems designed to monitor intein function, the 440-aa Mtu recA intein was reduced to a functional mini-intein of 137 aa. The accuracy of splicing of several mini-inteins was verified. This work not only substantiates structure predictions for intein function but also supports the hypothesis that, like group I introns, mobile inteins arose by an endonuclease gene invading a sequence encoding a small, functional splicing element.     

A mathematical model for the dynamic behavior of melts subjected to electromagnetic forces: Part I. model development and comparison of predictions with published experimental results

A mathematical model was developed to predict electromagnetically driven flow and (particularly) free surface behavior in melts subject to electromagnetic forces. Such melts appear in electromagnetic casters, induction furnaces, and other metal processing units. The calculations started with Maxwell’s equations and Ohm’s law, which were solved by a novel “modified hybrid technique.” The instantaneous continuity and Navier-Stokes equations (rather than their time-averaged versions) were then solved with electromagrretic forces as input. The calculations allowed for the dynamic behavior of the free surface of the melt, and electromagnetic fields were recomputed as the free surface changed. In this first part of a two-part article, the model predictions are compared with the experimental measurements of induced current, magnetic field, melt velocity, and free surface deformation reported by others.     

A mathematical model for the dynamic behavior of melts subjected to electromagnetic forces: Part II. Measurement of surface waves and comparison with predictions of the mathematical model

The waves on the surface of a cylindrical mercury pool subjected to a 3 kHz electromagnetic field have been observed. Oscillations were found to correspond to the known eigenfrequencies of a cylindrical liquid pool with a free surface. Which of the eigenfrequencies was dominant depended on the field strength and the relative position of the pool in the surrounding inductor. The amplitude of the oscillations was found to increase with increasing inductor current. The measurements showed fair to good agreement with the predictions of the mathematical model described in Part I. A tentative mechanism for the excitation of the eigenfrequencies by the electromagnetic field is advanced and implications for electromagnetic casting are discussed.     

Development of an animal model for neuroleptic malignant syndrome: heat-exposed rabbits with haloperidol and atropine administration exhibit increased muscle activity, hyperthermia, and high serum creatine phosphokinase level

The neuroleptic malignant syndrome (NMS) is a life-threatening complication of neuroleptic treatment. To elucidate the pathogenesis of NMS, an animal model has been developed. Experimental rabbits treated with haloperidol (1 mg/kg) by intramuscular injection, were studied for the diagnostic symptoms of increased muscle rigidity, elevated body temperature, and high serum creatine phosphokinase (CPK) level. Administration of haloperiodol (1 mg/kg) and atropine (0.4 mg/kg), and exposure to high ambient temperature (35 degrees C) induced a significant increase in electromyographic activity with muscle rigidity similar to that observed in patients with NMS. Such rabbits also showed elevated body temperature and serum CPK value. In addition to the similarity of the signs and symptoms, all parameters measured (muscle rigidity, body temperature, and serum CPK level) were normalized by dantrolene treatment. The effectiveness of dantrolene in the experimental animal partially confirms the validity of this animal model for NMS. This experimental animal model for NMS may be useful to elucidate the pathogenesis of NMS.     

Utilization of manpower: Development and evaluation of an assessment-classification model for matching individuals with jobs.

Describes a model for the assessment of individual characteristics, the identification of psychological requirements of jobs, and the classification of applicants to job opportunities. The validity of the model was investigated with 1,934 undergraduates. Subgroups, formed on the basis of previous behavioral data collected during the freshman year, differed with respect to criteria (e.g., arts-sciences vs applied studies and grade point average) measurements taken 4 yrs later. More importantly, the subgroups differed with respect to the curricular paths walked during college. Results indicate that it is possible to differentiate people in meaningful ways, to identify "job families," and to match people with jobs. (34 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The Royal College of Surgeons rat: an animal model for inherited retinal degeneration with a still unknown genetic defect

The Royal College of Surgeons (RCS) rat is the first known animal with inherited retinal degeneration. Despite the fact that the genetic defect is not known, the RCS rat is widely used for research in hereditary retinal dystrophies. This review tries to summarize observations which have been made in the RCS rat and to make an attempt to formulate candidate genes which may the cause for the retinal degeneration in this rat strain. The genetic defect in RCS rats causes the inability of the retinal pigment epithelium (RPE) to phagocytose shed photoreceptor outer segments. In normal rats or humans, this circadian process is regulated by both the cyclic adenosine monophosphate (cAMP) and the calcium/ inositol phosphate systems. The calcium/inositol phosphate system seems to be linked to the phagocytosis receptors which recognize photoreceptor outer membranes to initialize phagocytosis. The cAMP system appeared as modulator of the regulation of phagocytosis. An increase in the intracellular cAMP concentration is an 'off' signal for phagocytosis. In RPE cells from RCS rats many observations have been made which indicate a changed second messenger metabolism concerning both the cAMP and the calcium/inositol phosphate systems. The genetic defect seems to concern a protein which is involved in the initialization of a second messenger pathway. We conclude that the genes coding for the phagocytosis receptor or for proteins which are linked to receptors (for example G proteins) are good candidates for defective genes in RCS rats.     

A study of performance on tests from the CANTAB battery sensitive to frontal lobe dysfunction in a large sample of normal volunteers: implications for theories of executive functioning and cognitive aging. Cambridge Neuropsychological Test Automated Battery

Several tests from the CANTAB neuropsychological test battery previously shown to be sensitive to frontal lobe dysfunction were administered to a large group of normal volunteers (N = 341) ranging in age from 21 to 79 years. The main tests included a computerized form of the Tower of London test of planning, a self-ordered spatial working memory task, and a test of attentional set formation and shifting. A computerized form of the Corsi spatial span task was also given. Age-related graded declines in performance were seen, sometimes in a discontinuous manner, especially for the attentional set shifting task (at the extradimensional shift stage). Patterns of deficits reminiscent of frontal lobe or basal ganglia damage were observed in the oldest age group (74-79). However, overall the data were only partially consistent with the hypothesis that frontal lobe functions are the most sensitive to effects of aging. Factor analyses showed that performance in the executive tests was not simply related to a measure of fluid intelligence, and their performance had a factor loading structure distinct from that for the CANTAB tests of visual memory and learning previously administered to the same sample. Finally, only limited support was found for the hypothesis that cognitive aging depends on slowed information processing.     

Clinical trials among worker populations: a model for an anamnestic survey of upper limb pathologies and its practical application methods

Following a brief review of the principal clinical characteristics of musculoskeletal disorders of the upper limbs, the authors propose a protocol for a structured anamnestic examination featuring a series of set questions. The anamnestic model is based on a detailed listing of the symptoms to be analysed, which are divided into four categories: pain, paraesthesia, symptoms attributable to hyposthenia, and neurovegetative disorders. Regarding pain and paraesthesia, the authors list the localization, pattern of onset, duration and number of episodes, irradiation and treatment. The patients can thus be classified as anamnestic cases based on the following criteria: presence of pain or paraesthesia during the last 12 months, with episodes lasting for at least 1 week or occurring at least once a month, with no previous acute trauma. For hyposthenia, the authors report on the conditions under which the disorder may develop. The neurovegetative disorders considered are modifications in colour of the fingers and reaction to exposure to low temperatures. The structure of the proposed anamnestic chart permits all findings to be easily encoded for subsequent storage in a dedicated database. The appendix contains an annotated facsimile of the anamnestic chart.     

Emotions, coping and the need for support in families of children with cancer: a model for psychosocial care

Cloning and characterization of a Choristoneura fumiferana ultraspiracle (Cfusp) cDNA are described. First, a PCR fragment and then a cDNA clone (4.4 kb) were isolated from spruce budworm cDNA libraries. Comparison of the deduced amino acid sequence of this cDNA with the sequences in Genbank showed that this sequence had high homology with the ultraspiracle cDNAs cloned from Drosophila melanogaster (Dmusp), Bombyx mori (Bmusp), Manduca sexta (Msusp), and Aedes aegypti (Aausp). The Cfusp cDNA contained all the regions that are typical for a steroid/thyroid hormone receptor superfamily member. The DNA binding domain or C region was the most conserved sequence among all the usps. The A/B, D, and E regions also showed high amino acid identity with the amino acid sequences of Dmusp, Msusp, Bmusp, and Aausp. The Cfusp 4.5-kb mRNA was present in the embryos, in all larval stages, and in the pupae. The Cfusp mRNA levels in the midgut increased during the sixth-instar larval development and reached peak levels during the ecdysteroid raises for the pupal molt. However, Cfusp mRNA levels remained unchanged in the midgut of fifth-instar larvae, and in the epidermis and fat body of sixth-instar larvae indicating both a tissue- and stage-specific regulation of Cfusp mRNA expression.     

Young inflorescence-bearing shoots with roots of Tradescantia clone BNL 4430 cultivated in nutrient solution circulating systems: an alternative to potted plants and cuttings for mutagenicity tests

The use of young inflorescence-bearing shoots with roots of Tradescantia clone BNL 4430 cultivated in a nutrient solution circulating (NSC) growth chamber was tested and developed as an alternative method for using Tradescantia plants in mutagenicity testings. The NSC growth chamber was designed for our requirements, based on trial cultivations of the shoots with roots in its smaller-sized prototype. The nutrient solution used was a 1/2500 Hyponex solution. The characteristics of this clone, i.e., many new shoots constantly emerging from the basal nodes one after another and its short height favorable for early flowering, made it possible to prepare many young inflorescence-bearing shoots with roots at one time. A simplified NSC cultivation system could also be developed at a lower cost, and by using it together with the NSC growth chamber, recycling of untreated materials was established for supplying steadily enough amounts of young inflorescence-bearing shoots with roots for mutagenicity testings. Compared with traditional methods of using potted plants or cuttings, the new method exhibited more stable flower production, better stamen-hair growth and a significantly lower spontaneous (background) mutation frequency, and could produce more inflorescences per space. The use of such young inflorescence-bearing shoots with roots was therefore judged to be satisfactory to serve as a new mutagenicity test system alternating with potted plants and cuttings.     

Negative attributional style as a moderator of the link between perfectionism and depressive symptoms: Preliminary evidence for an integrative model.

An integrative model involving perfectionism and negative attributional style as predictors of depressive symptoms was proposed and tested in 222 college students. Beyond the expected additive influences of the 2 predictors in the prediction of dysphoria, negative attributional style was also hypothesized to moderate the link between perfectionism and depressive symptoms. As theorized, results indicated that dimensions of perfectionism (P. Hewitt & G. Flett, 1991b) predicted depressive symptoms (2 months later) and that a negative attributional style added incremental validity to these predictions. Moreover, consistent with the proposed model, the Perfectionism?×?Negative Attributional Style interaction was found to further augment the prediction of depressive symptoms. These findings are taken to offer preliminary support for the proposed model. Some implications for future studies are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Behavioral effects of phenelzine in an experimental model for screening anxiolytic and anti-panic drugs: correlation with changes in monoamine-oxidase activity and monoamine levels

This study investigated the effects of acute and chronic (one daily i.p. injection for 14 days) treatments with the non-selective irreversible monoamine-oxidase (MAO) inhibitor phenelzine (10 and 30 mg/kg) on defensive behaviors of Swiss mice in the mouse defense test battery (MDTB) which has been designed for screening anxiolytic and anti-panic drugs. In the MDTB, subjects were confronted with a natural threat (a rat) and situations associated with this threat. MAO-A and MAO-B activities and levels of brain monoamines (serotonin (5-HT), dopamine (DA) and norepinephrine (NE)) and their deaminated metabolites were subsequently measured. Behavioral results showed that acute administration of phenelzine did not specifically modify defensive behaviors. By contrast, after chronic treatment, phenelzine produced a significant reduction in avoidance distance when the rat was approaching, an effect which is consistent with an anti-panic-like action. In addition, phenelzine displayed weak anxiolytic-like effects as it increased risk assessment responses when mice were constrained in one part of the apparatus facing the rat which remained at a constant distance. No other specific drug effect was observed. These behavioral changes were associated with a dramatic increase in 5-HT levels, in particular after chronic treatment, while levels of DA and NE increased only slightly. Importantly, no significant differences in DA and NE levels between acute and chronic regimens were observed. Levels of deaminated metabolites of monoamines were markedly decreased. Measurements of MAO activity revealed substantial reductions in both type A and B forms with a full inhibition of both forms being observed only after chronic treatment with phenelzine. These results suggest that the effects of phenelzine may be due mainly to its effects on the 5-HT system and presumably related to the full inhibition of MAO-A and/or MAO-B.     

Location of induction and expression of protective immunity against Fasciola hepatica at the gut level: a study using an ex vivo infection model with ligated gut segments

PURPOSE: We developed a semiautomatic class solution to irradiate centrally located Stage III non-small cell lung cancer (NSCLC), involving a beam intensity modulation technique and optimization using a biophysical cost function. METHODS AND MATERIALS: Treatment for 10 patients with Stage III NSCLC was planned, using a conventional three- or four-beam three-dimensional (3D) technique and two techniques involving, respectively, seven (BIM1) and five (BIM2) noncoplanar beam incidences with intensity modulation. Two planning target volumes were defined: PTV1 included macroscopic tumor volume and PTV2 included macroscopic and microscopic disease. Beams were divided into beam parts (segments) and their outlines were defined during virtual simulation. Optimization using a biophysical cost function determined beam weights, segment weights, and wedge angles. Biological end points included tumor control probability of both target volumes (TCP1 and TCP2) and normal tissue complication probability (NTCP) of heart, lung, and spinal cord. The resulting uncomplicated local control probability (UCLP) was calculated. Physical end points included dose at PTV1 expressed as a dose minimum and dose maximum. Target-dose inhomogeneity was constrained in all plans. RESULTS: Concerning tumor evaluation, TCP1 was 74% (range 54-89%) for the 3D plan, 78.0% (range 62-94%) for BIM1, and 86.0% (range 59-93%) for BIM2. TCP1*TCP2 was, respectively, 67.0% (range 39-81%), 73.0% (range 56-94%), and 81.0% (range 54-93%). Minimum doses to PTV1 were 85, 80, and 88 Gy with the three respective techniques, while dose maxima were 89, 101, and 100 Gy. NTCPs of lung were 45.0% (range 11-75%) for 3D, 19.5% (range 8-59%) for BIM1, and 24.5% (range 3-61%) for BIM2. NTCPs of heart and spinal cord were comparable for all techniques. ULCPs were 37.0% (range 9-73%), 52.5% (range 22-86%), and 60.0% (range 20-85%), respectively. Applying physical limits to ensure clinical safety, minimum doses at PTV1 were recalculated. These were 72, 71, and 74 Gy for 3D, BIM1, and BIM2, respectively. CONCLUSION: The BIM2 plan is a candidate class solution for dose escalation studies in centrally located Stage III NSCLC.     

Familial coping with cancer in childhood and adolescence. Possibilities for standardized assessment with a self-assessment method: results of an empirical pilot study

44 parents of pediatric oncology patients answered mailed questionnaires about their own coping and about their sick children's coping and bodily complaints during oncological therapy. Most of the families coped well from parents' view. The dominant coping style of the parents consisted of active, problem-focused coping strategies in combination with an optimistic basic attitude, maintainance of family cohesion and belief in god or search for a meaning of the illness. The children and adolescents used predominantly adaptive coping strategies. The parental problem-group with a predominantly maladaptive coping-style reported low family cohesion, high depression, high wishful thinking and minimization. The maladaptive children and adolescents were rated as more socially withdrawing, more irritable, less optimistic and competent as well as less compliant. Strong correlations were found between parents' and their children's coping. Problem families with a special need for psychosocial support can early be identified by the coping questionnaires used in this study. Prospective longitudinal studies with multi-methodal and multi-perspective designs should be conducted to reach a better understanding of the family coping process in case of childhood and adolescence cancer.     

Method for establishing and measuring in vivo forces in an anterior cruciate ligament composite graft: response to differing levels of load sharing in a goat model

In order to determine the appropriate load history for optimal remodeling of an anterior cruciate ligament graft, methods for establishing and measuring the graft force due to an external load could be set to a preselected value in in vivo are required. Our objectives with this study were to (a) develop a method in which the graft force due to an external load could be set to a preselected value in a living animal, (b) show that this force could be maintained after fixation, and (c) determine what happens to the forces after the animal has functioned for as long as 2 weeks postoperatively, when differing levels of load sharing between the segments had been set at surgery. The anterior cruciate ligament was reconstructed in 12 goats with use of a bone-patellar tendon-bone graft and a synthetic augmentation device. The forces in the graft segments were established, at the time of surgical fixation, with use of a force-setting technique. In five animals, the tendon segment was set to carry 90% of the total graft force; in the remaining seven animals, the augmentation segment was set to share 90% of the total graft force. Graft forces were measured, with the use of buckle transducers mounted extra-articularly over the anterior tibia, under a 67 N anterior tibial load at 60 degrees of knee flexion before and after fixation and at 2 weeks postoperatively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chrome brightens the way for sinter-hardening: Combining sintering and heat-treatment in a single, clean, process step makes sinter-hardening an attractive and cost-effective option for the production of high-performance PM parts…

The economical achievement of premium mechanical properties for PM parts has been pursued over a number of years and a plethora of materials and process studies on sinter-hardening have already been launched. So it was not surprising that the sinter-hardening behaviour of ferrous PM materials was one of the major themes at the Las Vegas conference, writes Joe Capus.