Drosophila morphogenesis: orchestrating cell rearrangements.

Changes in shape of individual cells need to be coordinated to generate the movements of cell groups and sheets that are so important in morphogenesis. Recent results have shown that, during Drosophila gastrulation, multiple signalling pathways act to orchestrate the complex cell rearrangements.

The neural basis of ego- and allocentric reference frames in spatial navigation: evidence from spatio-temporal coupled current density reconstruction.

Different strategies in spatial navigation during passages through computer-simulated tunnels were investigated by means of EEG source reconstruction. The tunnels consisted of straight and curved segments and provided only visual flow, but no landmark, information. At the end of each tunnel passage, subjects had to indicate their end position relative to the starting point of the tunnel. Even though the visual information was identical for all subjects, two different strategy groups were identified: one group using an egocentric and the other group an allocentric reference frame. The current density reconstruction revealed the use of one or the other reference frame to be associated with distinct cortical activation patterns during critical stages of the task. For both strategy groups, an occipito-temporal network was dominantly active during the initial, straight tunnel segment. With turns in the tunnel, however, the activation patterns started to diverge, reflecting translational and/or rotational changes in the underlying coordinate systems. Computation of an egocentric reference frame was associated with prevailing activity within a posterior parietal-premotor network, with additional activity in frontal areas. In contrast, computation of an allocentric reference frame was associated with dominant activity within an occipito-temporal network, confirming right-temporal structures to play a crucial role for an allocentric representation of space.

Pressure variation of the lateral diffusion in lipid bilayer membranes.

A pressure-induced decrease of the lateral diffusion in pure and cholesterol containing phosphatidylcholine bilayer membranes has been determined by the excimer formation technique using pyrene as probe molecule. The experimental results at pressures up to 150 bars are described satisfactorily by the free volume theory of a molecular transport in liquids. A pressure increase of extrapolated 575 bars decreases the lateral diffusion of lipids by a factor of two in pure dipalmitoylphosphatidylcholine membranes. Higher pressures are necessary to induce the same effect in cholesterol containing membranes. This result is interpreted by the condensing effect of cholesterol in fluid bilayer membranes.

Lateral diffusion of small solutes and partition of amphipaths in defect structures of lipid bilayers.

The excimer formation technique has been applied to investigate the mechanism of the lateral diffusion in the crystalline P beta' phase of dipalmitoylphosphatidylcholine vesicles. This became possible at low pyrene concentrations. From the shape of phase-transition curves, from the effect of pressure up to 150 bar and from the perturbation induced by cholesterol, we conclude that the free volume model could also be applied to the P beta' phase. However, the diffusion is thought to occur in defect structures that are considered to form fluid pathways between domains of crystalline lipid. Partition coefficients of amphipaths provide a basis for testing for the role of defects. The amphipath chlorpromazine partitions into fluid membranes with a partition coefficient, kp, of 3200, into the crystalline phase with kp = 200 but into the P beta' phase with a value of kp = 800. This again gives rise to the assumption that the P beta' phase contains fluid domains that behave like the fluid L alpha phase and make up about 10-20% of the total amount of lipid in the bilayer. Cholesterol is known to interfere especially with defect structures in the P beta' phase. It fills up the gaps, and therefore reduces the partition coefficient to almost zero in the P beta' phase.

Exclusion of an extracolonic disease modifier locus on chromosome 1p33-36 in a large Swiss familial adenomatous polyposis kindred.

Familial adenomatous polyposis (FAP), an autosomal dominantly inherited colorectal cancer predisposition syndrome, displays considerable inter- and intrafamilial phenotypic heterogeneity, which represents a major problem in genetic counselling of APC mutation carriers. The Min mouse model indicated a putative disease modifier locus on chromosome 4, which is syntenic to human chromosome 1p35-36. This finding was subsequently supported by parametric and nonparametric linkage analyses in FAP families, however, without identifying functional variants in candidate genes. Recently, germline mutations in the base-excision repair gene MYH (1p33-34) have been described in patients with multiple adenomas, pointing to a possible role as disease modifier in FAP. Here, we present critical reassessment of one of the largest FAP kindreds published, which was previously used in linkage mapping of 1p35-36. In this family, all affected members harbour the same APC germline mutation (5945delA), but display marked phenotypic variability, in particular regarding the occurrence of extracolonic disease that segregates in several branches of the family tree. Using updated clinical information, additional mutation carriers and polymorphic markers, fine mapping of the critical region as well as mutation analysis of the MYH gene were performed. These investigations allowed us to significantly exclude (i) the 1p33-36 region as a modifier locus and (ii) MYH as a modifier gene for extracolonic disease in this FAP kindred. Our results do not eliminate 1p33-36 from suspicion in other families, but clearly indicate that in our family linkage analysis of further putative candidate regions is necessary to identify a disease modifier locus in FAP.

Synthetic human tRNA(UUULys3) and natural bovine tRNA(UUULys3) interact with HIV-1 reverse transcriptase and serve as specific primers for retroviral cDNA synthesis.

Full-length and 5'-truncated variants of human (h) tRNA(UUULys3) were synthesized by in vitro transcription using SP6 RNA polymerase. Bovine(b) tRNA(SUULys3) was purified from calf liver. Both full-length tRNA species were shown to be biologically active in an aminoacylation assay. Gel retardation assays revealed that both full-length tRNA species, as well as a 5'-truncated h-tRNA(UUULys3) molecule containing 24 nucleotides (nt) at the 3' end (Lys24), interact with human immunodeficiency virus (HIV)-1 reverse transcriptase (RT). Competition studies with these three tRNA species demonstrate that the 3' end of h-tRNA(UUULys3) contributes to the interaction with HIV-1 RT. Escherichia coli tRNA(UUULys) and tRNA(UUCGlu2) were also able to interact with the enzyme, whereas unrelated RNA molecules such as E. coli 5S rRNA did not bind to RT. Both b-tRNA(SUULys3) and h-tRNA(UUULys3) molecules, as well as the 5'-truncated variants, could be demonstrated to prime cDNA synthesis specifically using a HIV-1 RNA template, prepared by in vitro transcription, indicating that other viral or cellular proteins are not essential for this process. E. coli tRNA(UUULys) and tRNA(UUCGlu2), although able to interact with HIV-1 RT, failed to prime retroviral transcription. Products of cDNA synthesis were characterized by polymerase chain reaction, demonstrating that at least 18 nt at the 3' ends of h-tRNA(UUULys3) and b-tRNA(SUULys3) are still present in the cDNA product, whereas the 5' ends of both primer molecules were removed by the RNase H activity of HIV-1 RT.

Use of L-asparaginase in childhood ALL.

Owing to the high efficacy of L-asparaginase in the treatment of acute lymphatic leukaemia the enzyme was introduced into the chemotherapy schedules for remission induction of this disease shortly after results of large-scale clinical trials had become available. Since asparaginase monotherapy was associated with a high response rate but short remission duration, the enzyme is currently employed within the framework of combination chemotherapy schedules which achieve treatment response in about 90% and long-term remissions in the majority of patients. Recently initiated clinical trials have still confirmed the eminent value of asparaginase in the combination chemotherapy of acute lymphatic leukaemia and of some subtypes of non-Hodgkin lymphoma, and its important role as an essential component of multimodal treatment protocols. Despite the unique mechanism of action of this cytotoxic substance which shows relative selectivity with regard to the metabolism of malignant cells, some patients experience toxic effects during asparaginase therapy. Immunological reactions toward the foreign protein include enzyme inactivation without any clinical manifestations as well as anaphylactic shock. Severe functional disorders of organ systems result from the impaired homeostasis of the amino acids asparagine and glutamine. The changes affecting the proteins of the coagulation system have considerable clinical impact as they may induce bleeding as well as thromboembolic events and may be associated with life-threatening complications when the central nervous system is involved. Risk factors predisposing to thromboembolic complications are hereditary resistance against activated protein C and any other hereditary thrombophilia. Other organ systems potentially affected by relevant functional disorders are the central nervous system, the liver, and the pancreas, with patients who have a history of pancreatic disorders carrying an especially high risk of developing pancreatitis. Studies on the mechanisms of action and the occurrence of resistance phenomena have shown that a treatment response may only be expected if the malignant cells are unable to increase their asparagine synthetase activity to an extent providing enough asparagine to the cell; one may thus conclude that the enzyme-induced asparagine depletion of the serum constitutes the decisive cytotoxic mechanism. Independent of the asparagine depletion related cytotoxicity however, there are other mechanisms of clinical relevance like induction of apoptosis. Besides this, further influences on signal transduction cannot be excluded. Only few publications have dealt with the question of minimum trough activities to be ensured before each subsequent asparaginase dose in order to maintain uninterrupted asparagine depletion under treatment, and answers to this problem are not definitive. Clinical studies using enzymes from E. coli strains indicate that a trough activity of 100 U/l will suffice for complete asparagine depletion of the fluid body compartments with the preparations studied. These findings have been transferred to enzymes from other E. coli strains as well as those isolated from Erwinia chrysanthemi and to the PEG-conjugated E. coli asparaginases. It might be desirable to countercheck the results for confirmation or correction. The dosage and administration schedule of the various enzyme preparations required for complete asparagine depletion over a period of time have been insufficiently defined. While pharmacokinetic studies showed clinically relevant differences in biological activity and activity half-lives for enzymes from different biological sources, the findings of recently published clinical trials indicate that the therapeutic efficacy is affected when different asparaginase preparations are given by identical therapy schedules. (ABSTRACT TRUNCATED)

Longitudinal in vivo magnetic resonance imaging studies in experimental allergic encephalomyelitis: effect of a neurotrophic treatment on cortical lesion development.

Proton magnetic resonance imaging enables non-invasive monitoring of lesion formation in multiple sclerosis and has an important role in assessing the potential effects of therapy. T2-weighted and short tau inversion recovery magnetic resonance imaging were used to assess the effect of a neurotrophic adrenocorticotrophic hormone analogue [H-Met(O(2))-Glu-His-Phe-D-Lys-Phe-OH] on the volume of lesions in the brains of rats suffering from chronic experimental allergic encephalomyelitis, an animal equivalent of multiple sclerosis. Lesion volume was monitored during a five-month period. Magnetic resonance imaging indicated that treatment with the adrenocorticotrophic hormone analogue significantly reduced the lesion volume by 84 and 85% 10 and 20 weeks after lesion induction, respectively. Furthermore, peptide treatment significantly reduced chronic experimental allergic encephalomyelitis-related neurological symptoms during the chronic phase of the disease (week 3 until week 20 after lesion induction). Both functional and morphological recovery were considerably advanced by peptide treatment. Twenty weeks after lesion induction rats with chronic experimental allergic encephalomyelitis were killed for histological analysis, to correlate magnetic resonance imaging findings with morphological changes. The regions of abnormally high signal intensities on T2-weighted magnetic resonance images coincided with areas of demyelination and concomitant widespread inflammatory infiltration, oedema formation and enlarged ventricles. The improved neurological status and the 84% reduction in the lesion volume in the cerebrum of rats chronic experimental allergic encephalomyelitis point to the potential value of trophic peptides in the development of strategies for limiting the damage caused by central demyelinating lesions in syndromes such as multiple sclerosis.

Regression of left ventricular hypertrophy by AT1 receptor blockade in renal transplant recipients.

AT1 receptor antagonists control blood pressure (BP) effectively and reduce left ventricular hypertrophy in patients with essential hypertension. Because left ventricular hypertrophy is very common in renal transplant recipients, we examined the cardiovascular effects and the safety profile of the AT1 receptor antagonist losartan in hypertensive renal transplant recipients. In 20 renal transplant recipients with stable renal graft function 50 mg of losartan was added to the preexisting antihypertensive treatment (no angiotensin-converting enzyme inhibitors) at least 6 months after renal transplantation. Twenty-four-hour ambulatory BP, two-dimensional-guided M-mode echocardiography, and duplex sonography, as well as renal function, red blood cell count, cyclosporine A and FK 506 levels, erythropoetin, and angiotensin II concentration were determined at baseline and after 6 months of therapy. With 24-h ambulatory BP measurement, systolic blood pressure (SBP) was reduced by 7.5 +/- 2.4 mm Hg and diastolic blood pressure (DBP) by 4.5 +/- 1.8 mm Hg (P < .01 and P < .05, respectively). Posterior, septal, and relative wall thickness decreased by 0.95 +/- 0.2 mm, 0.91 +/- 0.2 mm and 0.04 +/- 0.01 mm, respectively (all P < .001). Left ventricular mass index decreased by 18.1 +/- 4.7 g/m2 (P < .01). Ejection fraction and midwall fractional fiber shortening as systolic parameters and the relation of passive-to-active diastolic filling of the left ventricle were unaltered. Serum creatinine and cyclosporine A concentration remained stable in all patients. Hemoglobin and hematocrit decreased by 1.0 +/- 0.3 g/dL and 3.6% +/- 0.9%, respectively (P < .002 and P < .001) without a change in serum erythropoetin level. In renal transplant recipients the AT1 receptor antagonist losartan reduces left ventricular hypertrophy without altering systolic or diastolic function. It is safe with regard to renal function and immunosuppression, but slightly decreases hemoglobin level.

Enhanced GABA(A) inhibition enhances synchrony coding in human perception.

The benzodiazepine, lorazepam enhances the efficiency of local, inhibitory GABA(A) synapses in the cortex, which stabilize postsynaptic, excitatory activity by synchronizing their own discharges at around 40 Hz. Treatment with lorazepam has also been shown to adversely influence detection performance in perceptual tasks, suggesting a role for GABA(A)-mediated synchronization during visuo-perceptual organization. Consistent with these findings we report that reaction times to target stimuli were slower following lorazepam treatment. However, when targets followed presentation of a synchronized prime, presented within a flickering 40-Hz display matrix, the effects of priming were amplified relative to baseline and control conditions. We conclude that enhanced GABA(A)-induced inhibition enhances stimulus-evoked synchronization with differential effects upon mechanisms of perceptual segmentation and grouping.

Mechanical lesions of the fimbria fornix in rat brain studied by 1H-magnetic resonance imaging. Evidence for long-lasting dynamic alterations in the ipsilateral ventricular system.

In vivo 1H-NMR imaging was employed to study dynamic changes in the status of tissue water as a function of time after mechanical brain injury induced by partial unilateral transection of the fimbria fornix (FF) in the rat brain and was correlated with histology. Changes in the brain tissue were reproducibly found in distinct regions which were exclusively located in the lesioned hemisphere. The most pronounced changes concerned the lateral ventricle. Ventricular enlargement became evident posterior to the site of transection after a few hours and was maximal after 2-4 days. At later time points the posterior ventricular expansion was reduced. The lateral ventricle anterior to the site of transection was significantly enlarged from day 1 and continued to expand for up to 7 months. Tissue response at the site of transection, mainly involving the hippocampal formation and the thalamus, was first manifested after 24 h, while signs of progressive tissue degeneration were apparent in the long term.

Rapid immunoturbidimetric assay of albumin and immunoglobulin G in serum and cerebrospinal fluid with an automatic discrete analyser.

A method is presented to determine albumin and immunoglobulin G (IgG) in serum and cerebrospinal fluid (CSF) with an automatic discrete analyser. Concentrations of albumin and IgG in serum and CSF are compared with values obtained by radial immunodiffusion (RID). Ratios of albumin in CSF over albumin in serum and the so-called IgG index are calculated. Ratios obtained with the discrete analyser and with the RID are compared and discussed with respect to clinical diagnosis.

Luminance-increment detection: capacity-limited or not?

Three experiments investigated whether spatial cuing influences luminance-increment detection accuracy. Ss saw multiple-target displays and responded yes or no to 4 locations, including cued position. To test whether cuing effects are due to the load on visual short-term memory from the number of locations, Experiments 1 and 2 presented displays with 4 or 8 relevant locations. Experiment 1 used peripheral cues; Experiment 2 used central cues. Significant cuing effects were less marked with 4- than 8-location displays. Cuing effects were largest with multiple targets, but a small reliable effect remained even with single targets. Experiment 3 replicated the single-target effect with predominantly multiple- and single-target displays. A capacity-limited selection account is developed for these findings and their implications are discussed for separate central and peripheral cuing mechanisms and the locus of spatial cuing effects.

Perceptual integration of motion and form information: is the movement filter involved in form discrimination?

Seven experiments reinvestigated J. Driver and P. McLeod's (1992) report of a "visual search asymmetry reversal" in a task that required the integration of motion and form information. They found that, when the form discrimination was easy, search was more efficient for a moving rather than a stationary conjunction target; the reverse was true when form discrimination was difficult. J. Driver and P. McLeod proposed that 2 mechanisms are involved: a "stationary form system," which supports accurate form discrimination but is relatively insensitive to movement, and a "movement filter," which segregates the moving from the stationary items but is relatively insensitive to aspects of form. The present experiments failed to find the asymmetry reversal. The results agree with the (more parsimonious) proposal that the function of the movement filter is limited to separating moving from stationary items, whereas form discrimination is accomplished within a unitary form system.

Reflexive and voluntary orienting of visual attention: time course of activation and resistance to interruption.

To study the mechanisms underlying covert orienting of attention in visual space, subjects were given advance cues indicating the probable locations of targets that they had to discriminate and localize. Direct peripheral cues (brightening of one of four boxes in peripheral vision) and symbolic central cues (an arrow at the fixation point indicating a probable peripheral box) were compared. Peripheral and central cues are believed to activate different reflexive and voluntary modes of orienting (Jonides, 1981; Posner, 1980). Experiment 1 showed that the time courses of facilitation and inhibition from peripheral and central cues were characteristic and different. Experiment 2 showed that voluntary orienting in response to symbolic central cues is interrupted by reflexive orienting to random peripheral flashes. Experiment 3 showed that irrelevant peripheral flashes also compete with relevant peripheral cues. The amount of interference varied systematically with the interval between the onset of the relevant cue and of the distracting flash (cue-flash onset asynchrony) and with the cuing condition. Taken together, these effects support a model for spatial attention with distinct but interacting reflexive and voluntary orienting mechanisms.

Probing distractor inhibition in visual search: inhibition of return.

The role of inhibition of return (IOR) in serial visual search was reinvestigated using R. Klein's (1988) paradigm of a search task followed by a probe-detection task. Probes were presented at either the location of a potentially inhibited search distractor or an empty location. No evidence of IOR was obtained when the search objects were removed after the search-task response. But when the search objects remained on, a pattern of effects similar to Klein's results emerged. However, when just the search-critical object parts were removed or when participants received immediate error feedback to prevent rechecking of the search objects, IOR effects were observed only when probes appeared equally likely at search array and empty locations. These results support the operation of object-based IOR in serial visual search, with IOR demonstrable only when rechecking is prevented (facilitating task switching) and monitoring for probes is not biased toward search objects.

Visual search for motion-form conjunctions: is form discriminated within the motion system?

Motion-form conjunction search can be more efficient when the target is moving (a moving 45 degrees tilted line among moving vertical and stationary 45 degrees tilted lines) rather than stationary. This asymmetry may be due to aspects of form being discriminated within a motion system representing only moving items, whereas discrimination of stationary items relies on a static form system (J. Driver & P. McLeod, 1992). Alternatively, it may be due to search exploiting differential motion velocity and direction signals generated by the moving-target and distractor lines. To decide between these alternatives, 4 experiments systematically varied the motion-signal information conveyed by the moving target and distractors while keeping their form difference salient. Moving-target search was found to be facilitated only when differential motion-signal information was available. Thus, there is no need to assume that form is discriminated within the motion system.

Visual search for conjunctions of motion and form: display density and asymmetry reversal.

In visual search for motion-form conjunctions, search rates have been reported to be faster for moving than for stationary targets if the target-nontarget discrimination is easy (45 degrees target line tilt from vertical), but this asymmetry is reversed if the discrimination is difficult (9 degrees tilt) (J. Driver & P. McLeod, 1992). Driver and McLeod proposed that gross aspects of form discrimination are performed within a motion filter that represents only the moving items, whereas fine discriminations rely on a stationary form system that is poor at filtering by motion. However, H. J. Müller and J. Maxwell (1994) failed to observe the asymmetry reversal, possibly because they used lower density displays. The study reported in this article also did not yield an effect due to varying display density. This lends support to the notion of a unitary form system, with the role of the motion filter being limited to guiding the search to the moving items or, if required by the task, the stationary items.

Dimension-based visual attention modulates dual-judgment accuracy in Duncan's (1984) one- versus two-object report paradigm.

Four experiments, adapting the object-judgment paradigm developed by J. Duncan (1984), examined the relationship between object-based and domain-based mechanisms of visual attention. The experiments demonstrated a cross-domain cost, in terms of accuracy, when observers made dual color-form judgments to one or two overlapping objects presented briefly, relative to within-domain, dual-color and dual-form judgments. This domain-based selection effect was additive to an object-based effect, a cost of making dual judgments to separate objects, as reported by J. Duncan (1984). The pattern of object- and domain-based effects points to a capacity limitation in how multidimensional features are bound into a coherent object representation, consistent with the dimension-weighting account of H. J. Miller, D. Heller, and J. Ziegler (1995), which postulates that there is a limit to the total selection weight available to be allocated to an object's dimensions.