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Biomedical subjects

L Lapointe

Publications and source records attributed to L Lapointe.

At least 19 recordsLinked to original sources

Evaluation of VITEK 2 for analysis of Enterobacteriaceae using the Advanced Expert System (AES) versus interpretive susceptibility guidelines used at Dynacare Kasper Medical Laboratories, Edmonton, Alberta.

OBJECTIVE: To evaluate the applicability and adaptability of the bioMérieux VITEK 2 Advanced Expert System (AES) to the customized interpretive susceptibility guidelines used at Dynacare Kasper Medical Laboratories (DKML). METHODS: Three hundred isolates of Enterobacteriaceae (not more than 30% Escherichia coli) were tested on the VITEK 2 system and the API 20E for identification. Susceptibility testing for these isolates was performed on the VITEK 2 system and the Pasco broth microdilution panels. Minimal inhibitory concentrations (MICs) and interpreted results according to the AES and DKML antimicrobial susceptibility guidelines were compared. RESULTS: Of 300 isolates tested for susceptibility, 13 did not give AES interpretations. Of the remaining 287 isolates, interpretations between AES and DKML guidelines were compared for 10 antibiotics. The overall correlation between the AES and DKML interpretations was 96.2%. CONCLUSION: This study demonstrated the benefits and limitations of the bioMérieux AES. Automated knowledge-based systems provide a useful laboratory tool for the interpretation of susceptibility results.

Alberta↗

Antibody response to an autogenous vaccine and serologic profile for Streptococcus suis capsular type 1/2.

An autogenous vaccine was developed, using sonicated bacteria, with a strain of Streptococcus suis capsular type 1/2. The objectives of this study were to evaluate the antibody response following vaccination and to assess the changes in antibody levels in pigs from a herd showing clinical signs of S. suis capsular type 1/2 infection in 6- to 8-week-old pigs. An enzyme-linked immunosorbent assay using the vaccine antigen was standardized. Results from a preliminary study involving 2 control and 4 vaccinated 4-week-old pigs indicated that all vaccinated pigs produced antibodies against 2 proteins of 34 and 43 kDa, respectively, and, in 3 out of 4 vaccinated pigs, against the 117-kDa muramidase-released protein. For the serologic profile, groups of 30 pigs from the infected herd were blood sampled at 2, 4, 6, 8, and 10 weeks of age. The lowest antibody level was observed between weeks 6 and 8, presumably corresponding to a decrease in maternal immunity. A marked increase was seen at 10 weeks of age, shortly after the onset of clinical signs in the herd. For the vaccination field trial, newly weaned, one-week-old piglets were divided into 2 groups of 200 piglets each (control and vaccinated); blood samples were collected from 36 piglets in each group at 2-week intervals for 12 weeks. A significant increase (P < 0.05) in antibody response was observed 4 weeks following vaccination and the level of antibodies stayed high until the end of the experiment. In the control group, the increase was only observed at 13 weeks of age, probably in response to a natural infection. The response to the vaccine varied considerably among pigs and was attributed, in part, to the levels of maternal antibodies at the time of vaccination. No outbreak of S. suis was observed in the control or vaccinated groups, so the protection conferred by the vaccine could not be evaluated.

Animals↗

Diethylstilbestrol regulates trophoblast stem cell differentiation as a ligand of orphan nuclear receptor ERR beta.

The orphan nuclear receptor ERR beta is expressed in undifferentiated trophoblast stem cell lines and extraembryonic ectoderm, and genetic ablation of ERR beta results in abnormal trophoblast proliferation and precocious differentiation toward the giant cell lineage. Here, we show that the synthetic estrogen diethylstilbestrol (DES) promotes coactivator release from ERR beta and inhibits its transcriptional activity. Strikingly, treatment of trophoblast stem cells with DES led to their differentiation toward the polyploid giant cell lineage. In addition, DES-treated pregnant mice exhibited abnormal early placenta development associated with an overabundance of trophoblast giant cells and an absence of diploid trophoblast. These results define a novel pathway for DES action and provide evidence for steroidlike control of trophoblast development.

Animals↗

Spatiotemporal dynamics of Floerkea proserpinacoides (Limnanthaceae), an annual plant of the deciduous forest of eastern North America.

Because environmental filters are temporally and spatially heterogeneous, there often is a lack of significant relationship between the spatial patterns of successive life stages in plant populations. In this study, we determined the spatiotemporal relationships between different life stages in two populations of an annual plant of the deciduous forests of eastern North America, Floerkea proserpinacoides. Demographic surveys were done over a 4-yr period, and experiments were performed in the field and under controlled conditions to test for the effects of various environmental factors on population dynamics. There was a general lack of relationship between the spatial patterns of seed bank and seedling density, and a lack of similarity between their spatial correlograms. This was related mostly to the effects of spatially variable environmental filters operating on germination and emergence. However, environmental filters acting on plant survival were stable through time and contributed to stabilize the density and spatial patterns of the populations. Despite density-dependent presenescence mortality, spatial patterns of seedlings and mature individuals were similar and their correlograms were alike, suggesting that mortality did not fully compensate for density. Estimated fecundity was negatively correlated with population density over the study period. Although flower production started only 2-3 wk after emergence, seed maturation mostly occurred at the end of the life cycle, just before the onset of plant senescence. Yet, individual fecundity was low for an annual plant, i.e., 3.0 ± 0.5 mature seeds/plant (mean ± 1 SE). Seed predation by vertebrates was not significant. Low soil moisture had little effect on the total number of seeds germinating, although it slowed down the germination process. In quadrats where leaf litter was experimentally doubled, seedling emergence was lower than in control quadrats; in quadrats where leaf litter was completely removed, emergence did not differ from that in control quadrats. Susceptibility to drought stress was higher for seedlings than for mature plants. Although the species does not maintain a long-term persistent soil seed bank, other factors, such as density-dependent fecundity and autogamy, may temper population fluctuations through time and reduce the probability of local extinction.

Journal Article↗

Fruit development in trillium. Dependence On stem carbohydrate reserves

Leaves are the main source of carbon for fruit maturation in most species. However, in plants seeing contrasting light conditions such as some spring plants, carbon fixed during the spring could be used to support fruit development in the summer, when photosynthetic rates are low. We monitored carbohydrate content in the rhizome (a perennating organ) and the aboveground stem of trillium (Trillium erectum) over the entire growing season (May-November). At the beginning of the fruiting stage, stems carrying a developing fruit were harvested, their leaves were removed, and the leafless stems were maintained in aqueous solution under controlled conditions up to full fruit maturation. These experiments showed that stem carbohydrate content was sufficient to support fruit development in the absence of leaves and rhizome. This is the first reported case, to our knowledge, of complete fruit development sustained only by a temporary carbohydrate reservoir. This carbohydrate accumulation in the stem during the spring enables the plant to make better use of the high irradiances occurring at that time. Many other species might establish short-term carbohydrate reservoirs in response to seasonal changes in growing conditions.

Journal Article↗

Cellular uptake and trafficking of a prototypical magnetic iron oxide label in vitro.

RATIONALE AND OBJECTIVES: Target-specific magnetic resonance (MR) contrast agents are being developed to improve the accuracy of MR imaging. The purpose of this study was to determine the mechanism of cell uptake, and modes of intracellular trafficking of a prototypical iron oxide label (RMA) used in the synthesis of some target-specific MR contrast agents. METHODS: The prototypical agent (RMA) consisted of a dextran-coated monocrystalline iron oxide that was modified with rhodamine (fluorescent label) and opsonized with albumin. Fluorescence microscopy was performed in a phagocytic C6 cell line and in murine bone marrow macrophages. Immunohistochemistry against lysosomal markers was used to confirm the intracellular location of the label. RESULTS: RMA was identified inside cells after incubation at concentrations as low as 4.0 x 10(-10) M Fe, typically observed with receptor mediated endocytosis and several orders of magnitude lower than that expected with fluid phase pinocytosis. Cell uptake could be inhibited by excess protein but not by dextran. RMA localized initially to tubular and to round intracellular organelles and co-localized with an antibody against a murine lysosomal glycoprotein antibodies (LGP-A) in macrophages. Three days after incubation, RMA was concentrated in perinuclear vesicles, which most likely represent terminal lysosomes where final breakdown appears to occur. CONCLUSIONS: The mechanism of cellular uptake of a prototypical opsonized iron oxide label is consistent with receptor-mediated endocytosis. Immediately after cell contact, RMA localizes to the lysosomal compartment and at long time points reside in vesicles that by morphology and distribution appear to be terminal lysosomes. Iron oxides therefore demonstrate metabolism via the lysosomal pathway.

Albumins↗

Arrangement of a cluster of three mouse type I keratin genes expressed sequentially during esophageal-type epithelial cell differentiation.

Keratins are intermediate filament proteins expressed in epithelial cells. They are divided into two groups, type I and type II, that must associate to form filaments. The genes encoding these proteins are clustered in two type-specific loci. In stratified epithelia, differentiation of the basal cells is accompanied by a switch in the expression of keratin genes. However, how this switch is controlled is not yet understood. We report here the cloning and mapping of a 55-kb region surrounding the keratin 19 (K19) gene in the mouse genome. This gene encodes a type I subunit expressed in simple and complex epithelia, notably in nonkeratinizing stratified epithelia of internal organs. In these tissues, it is expressed in basal cells and not in suprabasal cells, where the main type I subunit is keratin 13. Using probes corresponding to highly conserved sequences in intermediate filament proteins, we mapped two other genes downstream from the K19 gene. Restriction mapping and sequencing data indicate that they encode the mouse K15 and K13. The three genes are separated by about 5-6 kb, and they are in the same transcriptional orientation. Because the three genes are expressed together in stratified epithelia and because their order of expression during differentiation is the same as their order on the chromosome, we suggest that there is a relationship between their genomic organization and the control of their expression.

Amino Acid Sequence↗

Cell-specific transcription of the peripherin gene in neuronal cell lines involves a cis-acting element surrounding the TATA box.

Peripherin is a neurone-specific intermediate filament protein expressed mostly in the peripheral nervous system. To localize sequences that are important for the regulation of peripherin gene transcription, we have functionally dissected its promoter. Transfection into different cell lines and deletion mapping of peripherin-lacZ hybrid constructs indicated that the first 98 bp preceding the transcription start site of the gene were sufficient to confer cell-type specific expression. DNase I footprinting experiments revealed three protected sequences in this region, that were named PER1, PER2 and PER3. The PER2 and PER3 elements, localized between -98 to -46, interact with proteins that seem widely distributed. Deletion of these elements severely decreased the level of reporter gene activity. The PER1 element, which overlaps the TATA box, interacts with a DNA-binding protein prevailing in peripherin expressing cell lines. However, the core promoter, which contains the PER1 element, was inefficient in driving gene expression. Experiments designed to test the contribution of each element showed that PER2 and PER3 were important in determining the level of expression, while PER1 was important for cell-type specificity. In fact the polyoma virus enhancer linked to the peripherin gene core promoter was found to limit reporter gene activity to peripherin expressing cell lines. Together, these experiments indicate that co-operative interactions between different regions of the promoter are necessary for efficient and cell-type specific transcription of the peripherin gene in a subset of neuronal cells.

Animals↗

Resistance to low temperature photoinhibition is not associated with isolated thylakoid membranes of winter rye.

In vivo measurements of chlorophyll a fluorescence indicate that cold-hardened winter rye (Secale cereale L. cv Musketeer) develops a resistance to low temperature-induced photoinhibition compared with nonhardened rye. After 7.2 hours at 5 degrees C and 1550 micromoles per square meter per second, the ratio of variable fluorescence/maximum fluorescence was depressed by only 23% in cold-hardened rye compared with 46% in nonhardened rye. We have tested the hypothesis that the principal site of this resistance to photoinhibition resides at the level of rye thylakoid membranes. Thylakoids were isolated from cold-hardened and nonhardened rye and exposed to high irradiance (1000-2600 micromoles per square meter per second) at either 5 or 20 degrees C. The photoinhibitory response measured by room temperature fluorescence induction, photosystem II electron transport, photoacoustic spectroscopy, or [(14)C]atrazine binding indicates that the differential resistance to low temperature-induced photoinhibition in vivo is not observed in isolated thylakoids. Similar results were obtained whether isolated rye thylakoids were photoinhibited or thylakoids were isolated from rye leaves preexposed to a photoinhibitory treatment. Thus, we conclude that increased resistance to low temperature-induced photoinhibition is not a property of thylakoid membranes but is associated with a higher level of cellular organization.

Journal Article↗

The mouse keratin 19-encoding gene: sequence, structure and chromosomal assignment.

Keratin 19 (K19) is synthesized mainly in embryonic and adult simple epithelia, but has also been found in stratified epithelia as well. K19 is the smallest known keratin and is remarkable in that, contrary to all other keratins, it does not have a designated partner for the formation of filaments, implying that regulation of its expression is different from other keratin-encoding genes. As a first step in elucidating the mechanisms by which the K19 gene is regulated in relatively undifferentiated embryonic and in terminally differentiated adult tissues, a series of overlapping clones containing the complete mouse K19 gene was isolated from a mouse genomic library and characterized. The nucleotide (nt) sequence extends over 5119 nt and includes six exons. A region of 303 nt upstream from the transcription start point (tsp) was also sequenced. Comparison with the human and bovine K19 genes revealed the existence of homologies in both the coding and noncoding regions. The putative promoter region of the mouse K19 gene is highly homologous to the corresponding sequences of the human and bovine K19 genes. It contains an ATA box, a CAAT box and two potential Sp1-binding sites. Significant homologies were also found between the sequences of the introns of the mouse, human and bovine genes: this was particularly evident in introns 2, 3, 4 and 5. Intron 1, which showed the greatest degree of divergence, was found to contain many repetitive elements. Finally, it is shown that the mouse K19 gene cosegregates with the type-I keratin-encoding gene locus (Krt-1) on chromosome 11.

Amino Acid Sequence↗

Differential regulation of keratin 8 and 18 messenger RNAs in differentiating F9 cells.

F9 embryonal carcinoma cells (F9EC) can be induced to differentiate in vitro into epithelial cells expressing keratin 8 (K8) and keratin 18 (K18). cDNAs corresponding to K8 and K18 mRNAs were cloned and used to study the change in the abundance of these mRNAs during differentiation of F9 cells into parietal endoderm-like cells by treatment with retinoic acid (RA) or with RA and dibutyryl cAMP (Bt2cAMP). Using an RNase protection assay, it was determined that K8 mRNA was induced slightly before K18 mRNA and that it accumulated to a greater extent than K18 mRNA. Furthermore, differentiation in presence of Bt2cAMP plus RA resulted in an earlier induction of the two mRNAs and a higher level of expression of K8 mRNA. These results indicate that K8 and K18 mRNAs are regulated differently in F9 cells.

Animals↗

Effects of lung resection on pulmonary function and exercise capacity.

The effects of lung resection on exercise capacity and perception of symptoms were studied in 47 patients aged 39-73 (mean 58.3) years. Twenty had a pneumonectomy and 27 a lobectomy, all for lung cancer. Forced expiratory volume, maximal inspiratory and expiratory pressures, and progressive maximal one minute incremental cycle ergometer exercise performance were measured before and after surgery. Breathlessness and leg discomfort were assessed with a modified Borg scale (0-10). Mean FEV1 decreased from 79% (SD 22%) to 53% (11%) of the predicted value after pneumonectomy and from 89% (22%) to 74% (18%) after lobectomy. Exercise capacity, measured as the highest work load completed, Wmax, decreased from 78% (25%) to 58% (28%) predicted in the pneumonectomy group and from 77% (21%) to 67% (20%) in the lobectomy group. There was only a weak relation between changes in FEV1 and changes in Wmax (r = 0.54, r2 = 0.30). The slope of the relation between the intensity of dyspnoea and work load or the intensity of dyspnoea and ventilation increased significantly after pneumonectomy, but not after lobectomy. Leg discomfort increased more rapidly when related to work load after both pneumonectomy and lobectomy. After resection dyspnoea was rarely the only limiting factor at maximal exercise. It is concluded that (1) change in FEV1 is a poor predictor of change in exercise capacity after lung resection; (2) pneumonectomy results in a 25% decrease in Wmax and in an appreciable increase in dyspnoea during exercise; (3) lobectomy has little or no effect on Wmax or the intensity of postoperative dyspnoea; (4) after both pneumonectomy and lobectomy leg discomfort makes an important contribution to exercise limitation.

Adult↗

Differential expression of the epidermal K1 and K10 keratin genes during mouse embryo development.

Induction of genes coding for the K1 and K10 keratins during mouse development was studied by measuring the accumulation of their respective mRNAs in day 10 to 17 embryos using an RNase protection assay. Although these two keratins are coexpressed in the suprabasal layers of the epidermis, it was found that while K1 mRNA was detectable as soon as day 10, K10 mRNA was not detectable before day 12. The expression of these genes at this stage of development was not expected since they are specifically associated with keratinization, a process that does not begin before day 17 of gestation. Histological examination of the epidermis of day 10 to 17 embryos suggests that both genes are induced in cells committed to epidermal differentiation, after stratification has started but before the onset of keratinization. It was also found that the two mRNAs increased in abundance steadily and significantly until day 16 and that, in spite of the expectation that filaments should contain equivalent amounts of each subunit, K1 mRNA remained more abundant than K10 mRNA at all times including in adult epidermis. These observations indicate that the two genes are regulated independently during development.

Animals↗

Mouse keratin 19: complete amino acid sequence and gene expression during development.

The complete amino acid sequence of the mouse keratin 19 (K19) was determined from a partial sequence of cDNA isolated from a mouse (day 10.5) embryo library and an amplified genomic fragment. Analysis of the sequence reveals strong evolutionary conservation with other K19s. Examination of the expression of the gene encoding K19 (K19) during development using an RNase protection assay reveals it is expressed in extra-embryonic tissues by day 8.5 and in the embryo proper by at least day 9.5. Furthermore, the K19 gene is induced in differentiating F9 embryonal carcinoma cells. These results indicate that K19 is another keratin, in addition to the K8-K18 pair, which is synthesized early during mouse development. Finally, Southern analysis of the K19 gene reveals that it is found as a unique copy in the mouse genome, in contrast to what is found in humans, which have at least one processed pseudogene.

Amino Acid Sequence↗

Urorectodynamic evaluation of healthy volunteers.

Twelve female and 8 male healthy volunteers underwent urorectodynamic evaluation. Mean bladder capacity was 594 ml. and mean maximal vesical pressure 12.5 cm. water (H2O). Maximal bladder pressure correlated significantly with maximal mural tension (r = 0.96) but did not correlate well with bladder capacity (r = 0.20). Mean bladder volume at first desire to void was 32 per cent of mean bladder capacity. The bladder volume at the first desire did not correlate significantly with the capacity. Mean bladder pressure at 100 ml. volume was 2.9 +/- 0.4 cm. H2O (X +/- SEM). After bethanechol injection, the bladder pressure at 100 ml. volume increased by 8.8 +/- 1.2 cm. H2O (X +/- SEM) at twenty minutes. In only 1 subject did bladder pressure rise higher than 15 cm. H2O (5 per cent). Rectal pressure increased from 1.4 +/- 0.3 to 14.1 +/- 2 cm. H2O (X +/- SEM) at sixteen minutes. Electromyogram (EMG) of the external and sphincter did not follow regular pattern with vesical filling or attempt of voiding. The data obtained in these normal subjects may serve as a basis for comparison in the interpretation of data obtained in patients.

Adult↗

Evidences supporting an increased sympathetic tone and reactivity in a subgroup of patients with essential hypertension.

Several experimental evidences have shown that, under standarized conditions, circulating catecholamines (CA) or norepinephrine (NE) levels can be used as a valid index of the sympatho-adrenal activity in animal and man. This approach in the study of hypertensive patients has permitted to uncover that about 50% of patients with labile hypertension and about 30% of patients with stable hypertension had elevated CA levels at rest for 20 minutes in the supine position. The increased CA levels were mainly due to a rise in NE in stable hypertension and to a rise in epinephrine (E) in labile hypertension. On the basis of circulating CA levels, the hypertensive patients were divided into hyperadrenergic (CA levels above normal range) and normoadrenergic (CA levels within the normal range) subgroups. The hyperadrenergic labile or stable hypertensive subgroups were found to be also characterized by an enhanced CA or NE increase in response to change in position from supine to standing, by a faster heart rate and by an increased myocardial contractility, while these parameters were normal in the normoadrenergic subgroups. These findings support therefore the existence of an increased sympathetic tone and reactivity in association with hyperkinetic cardiac functions in an important population of hypertensive patients. In response to two weeks treatment with beta-blockers (either propranolol or metoprolol) hyperadrenergic stable hypertensive patients were found to be more responsive to this therapy than normoadrenergic patients although both groups had the same initial blood pressure. Moreover, this treatment lowered basal NE or CA levels and restored the enhanced CA or NE response to change in position toward normal in hyperadrenergic patients while it did not modify significantly circulating supine or standing CA and NE in normoadrenergic patients. These findings strongly support a participation of the sympathetic system in the maintenance of an elevated blood pressure in hyperadrenergic patients and raise the possibility of using a more rational approach in the therapy of hypertension.

Adult↗