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Biomedical subjects

A Mack

Publications and source records attributed to A Mack.

At least 37 records · Page 2Linked to original sources

Membrane properties of giant neurons in the caudal pontine reticular formation in vitro.

The intrinsic membrane properties of giant neurons of the caudal pontine reticular nucleus (PnC) were evaluated in a rat brain slice preparation. These neurons show low RP levels, long membrane time constants, low membrane resistance, tonic AP firing, and a lack of rectification behavior. The results are compared to those measured in adjacent areas of the reticular formation and discussed for their impact on triggering and modulating the acoustic startle response.

Animals↗

The Arabidopsis transposable element Tag1 is widely distributed among Arabidopsis ecotypes.

Tag1 is an autonomous transposable element (3.3 kb in length) first identified as an insertion in the CHL1 (NRT1) gene of Arabidopsis thaliana. Tag1 has been found in the Landsberg erecta ecotype of A. thaliana but not in Columbia or WS. In this paper, 41 additional ecotypes were examined for the presence of Tag1. Using an internal Tag1 fragment as probe, we found that DNA form 19 of the 41 ecotypes strongly hybridized to Tag1. Almost all of the Tag1-containing ecotypes had only one or two copies of Tag1 per haploid genome, as determined by Southern blot analysis. The only exception, Bf-1 from Bretagny-sur-Orge, France, had four copies. Two ecotypes, Di-G and S96, gave identical Southern blot patterns to that of Landsberg erecta and were subsequently shown to contain Tag1 at the same two positions found in Landsberg erecta (loci designated as Tag1-2 and Tag1-3). Two other ecotypes, Ag-0 and Lo-1, had a Tag1 element located at Tag1-2 but not at Tag1-3. The distance between these two loci was determined to be 0.37 cM. Analysis of DNA from two related species, A. griffithiana and A. pumila, showed that both species contain sequences that hybridize to Tag1 and that could be amplified with an oligonucleotide specific to the terminal inverted repeats of Tag1. These results show that Tag1 and related elements are present, and may be useful for insertional mutagenesis, in many A. thaliana ecotypes and several Arabidopsis species.

Arabidopsis↗

Butyrylcholinesterase antisense transfection increases apoptosis in differentiating retinal reaggregates of the chick embryo.

To investigate the roles of the enzymes butyryl- and acetylcholinesterase (BChE and AChE) in retinal proliferation and differentiation, we use reaggregated spheres from retinal cells of the 6-day-old chick embryo, forming cellular and fibrous areas homologous to all layers of a normal retina. Recently, we could suppress BChE expression by transfecting these so-called retinospheroids during their proliferation period with a pSVK3 expression vector containing a 5' fragment of the rabbit BChE gene in antisense orientation. Along with morphological changes, proliferation was significantly decreased. Here, we have studied the effect of antisense BChE suppression during the differentiation period of retinospheroids. As BChE is suppressed, the differentiation of AChE-positive cells is increased, whereas the immunoreactivities for red and green cone-specific opsins are strongly reduced. Concomitantly, the rate of apoptosis as determined by propidium iodide uptake, by increased CPP 32-like caspase expression, and by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling and DNA fragmentation assays is roughly doubled, predominantly at the expense of degenerating photoreceptor precursors. This is further strong evidence that the proliferation marker BChE regulates an intricate balance between cell proliferation, cell differentiation, and programmed cell death in this in vitro retinal system.

Acetylcholinesterase↗

Modification of glutamine synthetase expression by mammalian Müller (glial) cells in retinal organ cultures.

One of the key enzymes in glial-neuronal transmitter recycling is glutamine synthetase (GS). In the retina, GS is exclusively expressed by glial (Müller) cells where it serves to convert neuron-released active transmitter substances (glutamate and GABA) into glutamine. Experiments on avian retinae have shown that GS expression is developmentally regulated by glucocorticoid hormones and, to a lesser extent, by a non-hormonal control mechanism(s). Much less is known about GS regulation in mammalian retinae, although either increases or decreases of GS immunoreactivity have been observed in Müller cells in different forms of retinal pathologies. We studied GS expression in postnatal rabbit retinae both in vivo and explanted as wholemounts in vitro, using immunocytochemistry and Western immunoblotting. GS expression was detectable in vivo from the fourth postnatal day, and increased rapidly within the first weeks of life. Levels were lower in vitro than in vivo by an order of magnitude, and could be significantly stimulated (> 60-110%) in vitro by application of hydrocortisone, conditioned medium from cultured retinal pigment epithelium and glutamate or ammonia, but not GABA. It is concluded that GS expression in mammalian Müller cells is dependent on systemic control by glucocorticoid hormones, as observed in birds, but environmental (activity-dependent) factors may play a more important role in mammals.

Animals↗

Porcine lingual taste buds: a quantitative study.

BACKGROUND: Taste buds on the pig tongue have been neither studied in detail nor quantified. We have counted the numbers of fungiform and vallate papillae, estimated their taste bud densities, and computed numbers of taste buds per volume of papilla on porcine tongue. Pig was chosen for this work because it is an omnivorous animal, and is one of the better models for study of nutrition-related problems in omnivorous human beings. METHODS: Tongues from three adult pigs were obtained from the local abattoir, fixed in 10% neutral buffered formalin, and mapped for the fungiform and vallate papillae. Serial 15 microns sections were prepared from blocks containing the papillae, and were stained by H & E. Density and volume of the taste buds were estimated using a Bioquant system. RESULTS: Eight hundred fifty and 826 fungiform papillae were observed on tongue I and II, respectively; numerically vallate papillae were 1, 2, and 2 for tongue I, II and III, respectively. Number of taste buds per fungiform papilla was considerably less on the dorsal surface of the tongue (3.88/papilla) than on the lateral side (22.11/papilla). Vallate papillae contained significantly more taste buds, a mean of 732.67 per papilla (t = 14.27; df = 6; P = < 0.05), than in the fungiform papillae. Density of the buds per volume of fungiform papilla was notably less on the dorsum (13.69) than on the lateral border (30.08). The density of vallate papillae taste buds per volume of the papilla was significantly greater than the lateral and dorsal papillae taste bud density (t = 4.28; df = 6; P < or = 0.05). CONCLUSIONS: The present results are relevant to studies on variations of taste sensitivity among regions of the tongue.

Animals↗

Transfection of reaggregating embryonic chicken retinal cells with an antisense 5'-DNA butyrylcholinesterase expression vector inhibits proliferation and alters morphogenesis.

The function of the enzyme butyrylcholinesterase (BChE) in the developing and mature brain is still unclear. We have inserted 577 bp of the 5' upstream region plus 106 bp of the exon 1 of the rabbit BChE gene in reverse orientation under control of an SV40 early promoter derivative in an expression vector. This vector was introduced by calcium phosphate-mediated transfection into embryonic chicken retina cells during the first days of reaggregation culture. Depending on the retinal origin, the transfected cell population forms histotypic retina-like spheres, so-called rosetted or stratified retinospheroids. We show that antisense 5'-BChE gene expression decreased the steady-state mRNA level of BChE and the translation of the BChE protein, inhibited proliferation, and accelerated histogenesis in both cellular systems. The pronounced effects of antisense 5'-BChE transfection of spheroids document a key role of BChE during the early reaggregation process of retinal cells, most likely by regulating their growth and differentiation.

Animals↗

Admission examination factors predicting cognitive improvement during acute brain injury rehabilitation.

In this study 46 brain-injured patients admitted to a brain injury inpatient rehabilitation programme after acute care were evaluated at the bedside with a comprehensive mental status examination and physical evaluation. Using multiple regression analysis, elements of the cognitive and physical examinations were studied to determine if any of these items, individually or collectively, had predictive value for discharge functional status. A principal-components analysis identified meaningful clusters of items. Factor I, Simple Cognitive Operations, consisted of word repetition, naming and comprehension of simple commands. Factor IV, Higher Cognitive Operations, consisted of interpretation of similarities and reverse digit span. Other items, including elements of the physical examination such as eye movements, hearing and ambulatory function, were also clustered into factors. Stepwise multiple regression analysis led to a conclusion that only factor IV was a significant predictor of Discharge Rancho Level, with the predictive power of factor I and Admission Rancho Level subsumed by factor IV. This model, using a few items from a bedside mental status examination, accounted for 27% of the variance in Discharge Rancho Level in patients admitted to a brain injury rehabilitation unit after injury. We conclude that a brief mental status examination in a rehabilitation inpatient setting can be useful in predicting outcome in patients with brain injuries. More research is needed to identify additional test items that will add prognostic power to the initial evaluation of patients with brain injuries admitted to a rehabilitation unit.

Acute Disease↗

Regulation of cholinesterase gene expression affects neuronal differentiation as revealed by transfection studies on reaggregating embryonic chicken retinal cells.

In the embryonic chicken neuroepithelium, butyrylcholinesterase (BChE) as a proliferation marker and then acetylcholinesterase (AChE) as a differentiation marker are expressed in a mutually exclusive manner. These and other data indicate a coregulation of cholinesterase expression, and also possible roles of cholinesterases during neurogenesis. Here, both aspects are investigated by two independent transfection protocols of dissociated retina cells of the 6-day-old chick embryo in reaggregation culture, both protocols leading to efficient overexpression of AChE protein. The effect of the overexpressed AChE protein on the re-establishment of retina-like three-dimensional networks (so-called retinospheroids) was studied. In a first approach, we transfected retinospheroids with a pSVK3 expression vector into which a cDNA construct encoding the entire rabbit AChE gene had been inserted in sense orientation. As detected at the mRNA level, rabbit AChE was heterologously overexpressed in chicken retinospheroids. Remarkably, this was accompanied by a strong increase in endogenous chicken AChE protein, while the total AChE activity was only slightly increased. This increase was due to chicken enzyme, as shown by species-specific inhibition studies using fasciculin. Clearly, total AChE activity is regulated post-translationally. As an alternative method of AChE overexpression, transfection of spheroids was performed with an antisense-5'-BChE vector, which not only resulted in the down-regulation of BChE expression, but also strongly increased chicken AChE transcripts, protein and enzyme activity. Histologically, a higher concentration of AChE protein (as a consequence of either AChE overexpression or BChE suppression) was associated with an advanced degree of tissue differentiation, as detected by immunostaining for the cytoskeletal protein vimentin.

Animals↗

Stoichiometry of the Cre recombinase bound to the lox recombining site.

The site-specific recombinase Cre from bacteriophage P1 binds and carries out recombination at a 34 bp lox site. The lox site consists of two 13 bp inverted repeats, separated by an 8 bp spacer region. Both the palindromic nature of the site and the results of footprinting and band shift experiments suggest that a minimum of two Cre molecules bind to a lox site. We report here experiments that demonstrate the absolute stoichiometry of the Cre-lox complex to be one molecule of Cre bound per inverted repeat, or two molecules per lox site.

Base Sequence↗

Perceptual organization and attention.

It is widely assumed that the grouping of the visual field first described by the Gestalt psychologists and the related phenomenon of texture segregation occur very early in the processing of visual information and involve preattentive processes. All the recent evidence supporting this assumption comes from visual search experiments in which the subject is actively looking for a target and attending to the stimulus. The question at issue is whether these kinds of patterns are perceived under conditions of inattention, i.e., when observers are not searching for them. We performed six experiments to determine whether texture segregation and grouping by similarity or proximity are perceived under conditions of inattention. On the first two trials subjects were asked to report the longer arm of a briefly presented cross which was surrounded by a pattern of ungrouped small elements. On the third trial and subsequent control trials these elements were configured into grouping patterns and subjects queried about them immediately following their line length reports. The results establish that neither texture segregation nor grouping by similarity of lightness or proximity are perceived under conditions of inattention. They support the conclusion that there is an earlier stage of processing than that referred to as preattentive.

Adult↗

Perception without attention: results of a new method.

Having found by the use of a new method for examining perception without attention that grouping and texture segregation do not seem to occur (see Mack, Tang, Tuma, Kahn, & Rock (1992) Cognitive Psychology, 24, we go on to ask what is perceived without attention using this new method. Our subjects receive only one inattention trial in a sequence of trials involving a visual distraction task. In addition to the distraction task in the inattention trial, subjects received a stimulus of which they had no prior knowledge or expectation and were questioned or tested directly afterward for their perception of that stimulus. Two subsequent trials containing test stimuli serve as within-subject controls. The results of a series of experiments indicate that the presence of one or more stimulus objects and their locations are preattentively perceived, as is their color, but shape is not. Because individual items are detected without attention, we conclude that perceptual organization is initially based on a principle in which connected regions of uniform stimulation are inferred to be discrete units (the principle of uniform connectedness). One striking, unexpected finding is that without attention many subjects have no awareness at all of the stimulus object, an effect we call inattentional blindness.

Attention↗

Reversible menopause induced by the GnRH analogue buserelin: effects on lipoprotein metabolism.

GnRH agonists can induce a reversible pharmacological menopause and can be used to treat endometriosis, a fairly common gynaecological problem which responds well to oestrogen deprivation. Premature menopause is associated with adverse lipid changes and an increased risk of coronary heart disease. Therefore we set out to investigate changes in lipoprotein metabolism in a group of premenopausal women being treated with the GnRH analogue buserelin for active endometriosis. We monitored lipoprotein levels, high density lipoprotein subfractions and apolipoproteins AI, AII and B during a 12-month course of treatment and for 6 months afterwards. Treatment caused a sustained increase in HDL3 cholesterol levels and a small increase in low density lipoprotein cholesterol, which was significant at the six-month visit only. Apolipoprotein B levels rose significantly and there were marginal increases in apolipoproteins AI and AII. All changes and trends were reversed after cessation of treatment. We conclude that the treatment did not profoundly affect lipoprotein metabolism, neither LDL nor HDL cholesterol, the established important risk markers for coronary heart disease appreciably altered.

Adult↗

DNA specificity of the Cre recombinase resides in the 25 kDa carboxyl domain of the protein.

The Cre protein of bacteriophage P1 is a 38.5 kDa site-specific recombinase that belongs to the Int family of recombination proteins. Cre acts by binding specifically to a 34 base-pair sequence, lox, where it carries out recombination. A limited chymotryptic digest of Cre resulted in two fragments of sizes 25 and 13.5 kDa, respectively. The sequence of the amino terminus of the purified 25 kDa peptide demonstrates that this peptide represents the carboxyl-terminal portion of the Cre protein. A truncated version of the cre gene was constructed which produces only the 25 kDa peptide. The 25 kDa peptide is capable of specific binding to the lox site, but binds at lower affinity than does wild-type Cre. Footprinting with Fe-EDTA indicates that the 25 kDa peptide protects the inverted repeats of the lox site but shows only partial protection of the spacer region. This is in contrast to the footprint obtained with wild-type Cre which protects the entire spacer region.

Amino Acid Sequence↗

Motion aftereffects and retinal motion.

Two experiments are described in which it was investigated whether the adaptation on which motion aftereffects (MAEs) are based is a response to retinal image motion alone or to the motion signal derived from the process which combines the image motion signal with information about eye movement (corollary discharge). In both experiments observers either fixated a stationary point or tracked a vertically moving point while a pattern (in experiment 1, a grating; in experiment 2, a random-dot pattern) drifted horizontally across the field. In the tracking condition the adapting retinal motion was oblique. In the fixation condition it was horizontal. In every case in both conditions the MAE was horizontal, in the direction opposite to that of pattern motion. These results are consistent with the hypothesis that the adaptation is a response to the motion signal derived from the comparison of eye and image motion rather than to retinal motion per se. An alternative explanation is discussed.

Adaptation, Ocular↗

Apparent motion: evidence of the influence of shape, slant, and size on the correspondence process.

Four stimulus elements configured as a notional diamond were flashed in pairs to elicit apparent motion. When the elements were identical (4 Zs), the direction of apparent motion was ambiguous. When the elements were pairs of different letters (Cs and Os, Es and Zs), letters of different sizes (Zs and zs), or oppositely oblique lines, the direction of apparent motion tended to be between identical elements. This was true, however, only for an initial, brief observation period. Subsequently, the direction of apparent motion tended to be determined by the direction of motion perceived at first, regardless of the character of the elements. This quickly established directional set (within 10 sec) largely swamped any tendency to resolve correspondence in terms of a feature of the stimulus. It appears to be based on spatial rather than retinal or egocentric coordinates.

Attention↗