Slow cellular dynamics in MDCK and R5 cells monitored by time-lapse atomic force microscopy

We have examined dynamic events that occur on a time scale of minutes in an epithelial monolayer of Madine-Darby Canine Kidney (MDCK) cells and in ras-transformed MDCK cells by atomic force microscopy (AFM). Cells were imaged under physiological conditions, and time-lapse movies representing approximately 60 s real time per frame were assembled. In normal MDCK cells, two types of protrusions in the apical plasma membrane exhibit dynamic behavior. First, smooth bulges formed transiently over the time scale of minutes to tens of minutes. Second, spike-like protrusions appear initially as bulges, extend well above the apical surface and, finally, seem to detach. R5, an oncogenic transformant derived from MDCK cells, grows very flat on glass. During AFM imaging, these cells sometimes round up and detach from the substrate. In light microscopic observations of parallel preparations, cells rarely detach, suggesting that this is an active response of these cells to irritation by the AFM tip. R5 cells often extend processes that are supported by actin stress fibers. During imaging with the AFM, these processes withdraw at a rate of 1-5 microns/min, similar to that observed by light microscopy. During the withdrawal, movement of the stress fibers can be clearly seen. In the flat periphery of these cells, the transport of intracellular particles along cytoskeletal elements was seen. In addition, we have observed two types of wave-like movements through the cell, which appear to be an organized rearrangement of cytoplasm. One type of wave moves radially out from center of the cell while the other moves circularly along the cell periphery.     

Marfan syndrome: diagnosis of cardiovascular manifestations]

BACKGROUND: Despite the availability of several different markers for Epstein--Barr virus (EBV) serology, the EBV status of some patients cannot be resolved from a single serum sample with routine testing. To avoid the requirement of follow-up samples, supplementary tests have to be used in these cases. OBJECTIVE: To evaluate the usefulness of avidity and immunoblot assays as supplementary tests for the diagnosis of acute EBV infections. STUDY DESIGN: Three groups of samples for which a definite diagnosis on the EBV status could not be obtained with the routine serological tests were further examined by an EBV IgG avidity assay, by an immunoblot based on a lysate of EBV infected cells, and by a second immunoblot based on recombinant EBV antigens. The three groups consisted of 38 samples with negative/borderline EB nuclear antigen 1 (EBNA-1) antibodies, negative/borderline EBV IgM and positive EBV IgG; 10 samples with indeterminate EBNA-1 and/or EBV IgM assays because of control antigen reactions; and 4 samples with positive EBV IgM results that were not plausible. RESULTS: The avidity assay differentiated between acute and past infections for all samples. In contrast, some cases remained unresolved with both the recombinant and the lysate immunoblot. Two samples were incorrectly classified with the lysate immunoblot. Interpretation of the lysate immunoblot banding patterns was complicated when anticellular antibodies were present. CONCLUSION: Avidity testing appears to be the confirmatory method of choice to differentiate between acute and past EBV infections.     

Hemispheric contributions to the integration of visual and auditory information in speech perception

Differential hemispheric contributions to the perceptual phenomenon known as the McGurk effect were examined in normal subjects, 1 callosotomy patient, and 4 patients with intractable epilepsy. Twenty-five right-handed subjects were more likely to demonstrate an influence of a mouthed word on identification of a dubbed acoustic word when the speaker's face was lateralized to the LVF as compared with the RVF. In contrast, display of printed response alternatives in the RVF elicited a greater percentage of McGurk responses than display in the LVF. Visual field differences were absent in a group of 15 left-handed subjects. These results suggest that in right-handers, the two hemispheres may make distinct contributions to the McGurk effect. The callosotomy patient demonstrated reliable McGurk effects, but at a lower rate than the normal subjects and the epileptic control subjects. These data support the view that both the right and left hemisphere can make significant contributions to the McGurk effect.     

Mechanical properties of moist agglomerates in relation to granulation mechanisms part I. Deformability of moist,densified agglomerates

The mechanical properties of moist samples of fine, polydisperse powders are investigated by measuring the tensile strength by a diametrical compression test and the stress—strain relationship of samples exposed to uniaxial compression. The strengths determined by the two methods correlate well. The strength of moist samples compressed to porosities comparable to the level of intragranular porosities achieved by granulation in high-speed mixers is shown to be influenced significantly by particle interactions in addition to mobile liquid bondings. For a particular material the strength is controlled mainly by porosity and liquid saturation. It is shown that the effect of a growing liquid saturation is to reduce particle interactions and thus to facilitate densification during granulation. The deformability of moist samples, which is dependent on strength and deformation behaviour, is assessed for lactose and dicalcium phosphate.     

Mechanisms of cell damage and enzyme release

Release of intracellular enzymes to the extracellular space is a marker of cell damage in various diseases, e.g. liver, heart and muscle diseases. In the normal state the plasma membrane is impermeable to enzymes, and enzyme release, therefore, indicates a severe change of the membrane integrity. This review deals with the present knowledge about cellular changes leading to enzyme release, which may be caused either by energy depletion, e.g. in ischemia or shock, or by a direct membrane damage as caused by various toxins and inflammatory products. Inhibition of the energy metabolism results in ATP depletion leading to fluxes of Na+, K+ and Cl- down their gradients across the membrane and swelling of the cell. Subsequently Ca2+ leak into the cell activating phospholipases and the formation of eicosanoids, affecting the cytoskeleton and, perhaps, activating the formation of oxidants. The exact "point of no return" is not known but an uncontrolled Ca2+ activity in the cell probably has an important role in initiating the irreversible changes. The result of these reactions and probably other unknown reactions as well is damage to the membrane. This is evident morphologically at first by the formation of blebs that appears in the reversible phase, and later on by rupturing of the membrane, a sign of irreversible damage. A very small part of the enzyme release may occur in the reversible phase when blebs detach with resealing of the membrane, but the substantial part of enzyme release occurs as a result of irreversible cell damage when ATP has decreased to a low level and a serious disruption of the membrane integrity has taken place. All the secondary affections of the membrane during energy depletion may also occur as a primary direct membrane damage that more or less may affect the energy metabolism secondarily. The cell damage and enzyme release after some types of direct membrane damage is almost independent of the cellular energy metabolism whereas other types of direct membrane damage are counteracted by the cell by energy consuming reactions and, therefore, the final cell damage is a concerted action of the direct membrane damage and the energy depletion. This also means that a direct membrane damage may be more severe for the cell in energy depleted states than in the normal state. As in energy dependent cell damage the substantial part of enzyme release after a direct membrane damage is due to irreversible cellular changes. It appears that although the knowledge of the molecular basis of cell damage and enzyme release has grown there are still many questions to be answered about these complex processes.     

Log-linear allometry of fetal craniofacial growth in Down's syndrome

Trisomy 21 develops as a result of nondisjunction of two homologous chromosomes during either the first or second meiotic division. One of the more important consequences of these genetic alterations is the predictable, although variable disturbance in the architecture of the craniofacial region [1]. Postnatal craniofacial morphology has been extensively studied in Down's syndrome (DS). However, little information is available on human prenatal development of the head and face in such patients. The time at which changes in craniofacial phenotype first emerge in Down's syndrome fetuses and at which physical growth begins to diverge from normal is unknown. To explore these questions, we compared prenatal craniofacial growth in 50 Down's syndrome fetuses with that of 555 fetuses judged to be "typical for body weight and age" using the method of log-linear allometry [2].     

Surgical management of total anomalous pulmonary venous drainage: impact of coexisting cardiac anomalies

BACKGROUND: Recent reports have cited improving results for surgical management of isolated total anomalous pulmonary venous drainage. Complex cases (with other cardiac anomalies) are less frequently reported and are associated with higher mortality. METHODS: Retrospective review identified 170 consecutive patients treated for total anomalous pulmonary venous drainage from 1982 to 1996: 44 cases were "complex" (with significant associated cardiac lesions) and 126 cases were "simple." RESULTS: Operative mortality for simple cases decreased from 26% to 8%, and mortality for complex cases remained constant at 52%. Age, size, and the presence of atrial isomerism were univariate predictors of mortality. Multivariable analysis identified only univentricular hearts and associated cardiac lesions as predictors of operative mortality. Pulmonary artery (n = 16) and arteriopulmonary (n = 7) shunting strategies for complex cases resulted in less than 30% long-term survival. CONCLUSIONS: Despite improvement in survival for simple cases, management of total anomalous pulmonary venous drainage with single-ventricle hearts or other associated cardiac lesions remains problematic.