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Biomedical subjects

M Rougier

Publications and source records attributed to M Rougier.

At least 19 recordsLinked to original sources

Ultrasound-guided coxofemoral arthrocentesis in horses.

REASON FOR PERFORMING STUDY: Coxofemoral joint pain is probably underestimated due to difficulties in identifying hip pain. The deep location of the joint and proximity of the sciatic nerve make arthrocentesis based on external landmarks a difficult and potentially risky procedure in mature horses. OBJECTIVES: To describe an ultrasound-guided injection technique of the coxofemoral joint in standing horses and to evaluate its accuracy and potential difficulties/complications. METHODS: Nine mature horses had both pelvic areas prepared for sterile ultrasound examination (3.5 MHz curvilinear probe). Coxofemoral joints were located and penetrated at their craniodorsolateral aspect under ultrasonographic guidance and injected with sterile contrast medium. A standing ventrodorsal radiographic view of each hemipelvis centred on the hip was obtained for each horse to assess the injection site. Horses were evaluated for 10 days following injection for possible complications. RESULTS: Intra-articular injection was successful in all 18 joints. The procedure was well tolerated by horses under minimal restraint. Mean +/- s.d. needle repositionings required before accurate placement was 1.5 +/- 1.3 per joint. Once the needle was in the joint, synovial fluid was obtained in 7/18 joints. Minimal periarticular contrast medium was detected in 2/18 joints. Mean +/- s.d. ultrasonographic examination time required for coxofemoral localisation, accurate needle positioning and injection was 4.3 +/- 2.1 min. No complications were observed in the 10 days following injection. CONCLUSION: The ultrasound-guided coxofemoral arthrocentesis is an accurate, reliable and safe technique that offers a real time evaluation of needle introduction into the deep and narrow coxofemoral joint space. POTENTIAL RELEVANCE: Although this technique remains to be tested on clinical cases, it is a promising tool to facilitate diagnosis of coxofemoral pain, septic arthritis or administration of intra-articular medication.

Animals↗

Clinical isolate of vancomycin-heterointermediate Staphylococcus aureus susceptible to methicillin and in vitro selection of a vancomycin-resistant derivative.

A Staphylococcus aureus strain with low-level heteroresistance to vancomycin (designated MER) but susceptible to methicillin was isolated from an outpatient with conjunctivitis who did not receive any glycopeptide antibiotics. Incubation of the parent strain, MER, with increasing concentrations of vancomycin led to rapid selection of a stable progeny homogeneously resistant to vancomycin. Electron micrographs of strain MER showed enhanced cell wall thickness and abnormal septations typically seen with methicillin-resistant S. aureus having intermediate susceptibility to vancomycin.

Adult↗

A calcium influx is triggered and propagates in the zygote as a wavefront during in vitro fertilization of flowering plants.

In this paper, we report direct measurement of an influx of extracellular Ca(2+) induced by gamete fusion in flowering plants. This result was obtained during maize in vitro fertilization with the use of an extracellular Ca(2+)-selective vibrating probe. Ca(2+) influx recorded at the surface of isolated egg cells, with or without adhesion of a male sperm cell, was close to zero and stable over time. Gamete fusion, however, triggered a Ca(2+) influx in the vicinity of the sperm entry site with a delay of 1.8 +/- 0.6 sec. The Ca(2+) influx spread subsequently through the whole egg cell plasma membrane as a wavefront, progressing at an estimated rate of 1.13 micrometer.(-1). Once established, Ca(2+) influx intensities were sustained, monotonic and homogeneous over the whole egg cell, with an average peak influx of 14.92 pmol .cm(-2).(-1) and an average duration of 24.4 min. The wavefront spread of channel activation correlates well with the cytological modifications induced by fertilization, such as egg cell contraction, and with the cytosolic Ca(2+) ((c)[Ca(2+)]) elevation previously reported. Calcium influx was inhibited effectively by gadolinium, possibly implicating mechanosensitive channels. Furthermore, artificial influxes created by incubation with Ca(2+) ionophores mimicked some aspects of egg activation. Taken together, these results suggest that, during fertilization in higher plants, gamete membrane fusion starts the first embryonic events by channel opening and Ca(2+) influx. In turn, (c)[Ca(2+)] may work as a trigger and possibly a space and time coordinator of many aspects of egg activation.

Calcium↗

Genes normally expressed in the endosperm are expressed at early stages of microspore embryogenesis in maize.

Reproduction in flowering plants is characterized by double fertilization and the resulting formation of both the zygotic embryo and the associated endosperm. In many species it is possible to experimentally deviate pollen development towards an embryogenic pathway. This developmental switch, referred to as microspore embryogenesis or androgenesis, leads to the formation of embryos similar to zygotic embryos. In a screen for genes specifically expressed during early androgenesis, two maize genes were isolated by mRNA differential display. Both genes represent new molecular markers expressed at a very young stage of androgenic embryogenesis. When their expression pattern was studied during normal reproductive development, both showed early endosperm-specific expression. Investigation of the cytological features of young androgenic embryos revealed that they present a partially coenocytic organization similar to that of early endosperm. These findings suggest that maize androgenesis may possibly involve both embryogenesis and the establishment of endosperm-like components.

Blotting, Southern↗

[Third degree atrio-ventricular block induced by interferon alpha. Report of a case].

Iatrogenic third degree atrioventricular block due to alpha interferon is rare. The authors present a case which occurred with low dosage, regressed when treatment was withdrawn and reappeared when treatment was reintroduced. The physiopathological mechanism of disease of the conduction pathways and its general cardiotoxicity is not yet understood. The secondary effects of this increasingly widely used anti-tumoral and anti-infectious drug should be recognised in order to prevent them. Initial cardiological investigation and follow-up are indicated to ensure this prevention.

Cardiomyopathies↗

Inactivation of conidia of Paecilomyces fumosoroseus by near-ultraviolet (UVB and UVA) and visible radiation.

The detrimental effects of solar radiation, especially the ultraviolet waveband, on quiescent conidia of Paecilomyces fumosoroseus were investigated. Conidia were irradiated by a high-intensity source, which emitted a continuous spectrum from 270 to 1100 nm and which was equipped with long-pass filters to block short wavelengths below 280, 295, 320, or 400 nm. After irradiation, conidia were tested for germinability, survival, and infectivity toward Spodoptera frugiperda larvae. It was demonstrated that the detrimental effects of light depended on irradiance in the shortest wavelengths. The UVB (280-320 and 295-320 nm) appeared to be the most detrimental part of natural radiation, although UVA (320-400 nm) was also harmful. Visible and near infrared radiations were less harmful than UV. Our results demonstrate that the irradiance of the UVB waveband should be considered as the pertinent factor for the detrimental effects of sunlight on the persistence of conidia of entomopathogenic fungi in insolated environments.

Paecilomyces↗

B chromosome behavior in maize pollen as determined by a molecular probe.

The B chromosomes of maize typically undergo nondisjunction during the second microspore division (generative cell division). When the microspore nucleus contains only one B chromosome, two kinds of sperm result, one with two B chromosomes and one with no B chromosomes. The sperm with the B chromosomes preferentially fertilizes the egg cell. Previous studies of these phenomena have been limited to genetic analysis and chromosome spreads. In this study we show that a B chromosome-specific probe can be used with fluorescence in situ hybridization (FISH) analysis to detect the presence, location, and frequency of B chromosomes in intact interphase nuclei within mature pollen of maize. Using genetic line TB-10L18, our results indicate that nondisjunction of the B centromere occurs at an average frequency of 56.6%, based on four plants and 1306 pollen grains analyzed. This is consistent with the results of genetic studies using the same B-A translocation. In addition, our results suggest that B chromosome nondisjunction can occur during the first microspore division. Spatial distribution of the B chromosome-specific probe appears to be largely confined to one tip of the sperm nucleus, and a DNA fragment found outside the pollen nuclei often hybridizes to the B chromosome-specific probe.

Biotinylation↗

First evidence of a calcium transient in flowering plants at fertilization.

We report here the first evidence of a transient elevation of free cytosolic Ca2+ following fusion of sperm and egg cell in a flowering plant by the use of an in vitro fertilization system recently developed in maize. Imaging changes in cytosolic Ca2+ at fertilization was undertaken by egg cell loading with the fluorescent Ca2+ indicator dye fluo-3 under controlled physiological conditions. The gamete adhesion step did not induce any cytosolic Ca2+ variation in the egg cell, whereas the fusion step triggered a transient cytosolic Ca2+ rise in the fertilized egg cell, lasting several minutes. This rise occurred after the establishment of gamete cytoplasm continuity. Through these observations, we open the way to the identification of the early signals induced by fertilization in flowering plants that give rise to the calcium transient and to investigations of the role of Ca2+ during egg activation and early zygote development in plants, as has been reported for other better characterized animal and algae systems.

Aniline Compounds↗

Isolated maize zygotes mimic in vivo embryonic development and express microinjected genes when cultured in vitro.

We established conditions for the regeneration of natural maize zygotes isolated from pollinated plants with the goal of investigating the molecular control of early embryogenesis in higher plants. Viable zygotes were excised from embryo sacs by minimal enzymatic digestion and microdissection. Viable zygotes transferred to coculture with androgenic microspores from barley developed into embryo-like structures in 61% of the cases. No development was observed when zygotes were cultured in the presence of maize anthers undergoing androgenetic embryogenesis. Zygote-derived embryo-like structures regenerated into fertile plants through secondary embryogenesis when transferred to solid medium. The first zygotic division was asymmetrical and bipolar structures similar to pretransitional embryos observed in planta were later produced as observed using light and electron microscopy. Conditions for efficient microinjection of DNA into zygotes were established. Calcofluor and PATAG staining of zygotes showed that cell wall regeneration occurred as early as 20 min after enzymatic isolation and that after 2 hr, each zygote was bordered with cell wall material. Through quantitative microphotometry, DNA synthesis during the first cell cycle of the zygote was shown to occur between isolation and 12 hr of culture. Microinjection of two types of reporter genes (GUS gene and anthocyanin regulatory genes) demonstrates transient expression in plant zygotes. On average, 3.5% of microinjected zygotes showed transgenic expression. Reporter gene expression was observed in zygotes at different time points of their first cell cycle.

Cell Cycle↗

[Kinetics of in vitro bactericidal activity of the antiseptic biseptine combining 3 active principles].

Biseptine, is an association of chlorhexidine digluconate, benzalkonium chloride and benzylic alcohol. Little is known, in literature, on this antiseptic, used for cutaneous antisepsis. We studied killing kinetic of Biseptine, at different concentrations, on 4 AFNOR bacterial strains. Killing curves were studied at antiseptic concentration of 90 to 0.1% in 10 ml of distilled water. Bacterial counts were determined after neutralization in liquid medium. Synergy of chlorhexidine and benzalkonium chloride in Biseptine, allowed to obtain similar bactericidal activity than Hibitane champ with chlorhexidine concentrations 2 fold less. At 90, 50, 25, 10 and 5% concentrations, bactericidal activity (5 log10 reduction of the initial bacterial count) was effective in one minute. After 5 to 15 minutes, activity persisted at 1 and 0.5% concentrations. The 0.1% solution was inefficacious. This report disclosed an important security margin in antiseptic activity.

Anti-Infective Agents, Local↗

Expression level of the SLG gene is not correlated with the self-incompatibility phenotype in the class II S haplotypes of Brassica oleracea.

In Brassica, the S-locus glycoprotein (SLG) gene has been strongly implicated in the self-incompatibility reaction. Several alleles of this locus have been sequenced, and accordingly grouped as class I (corresponding to dominant S-alleles) and class II (recessive). We recently showed that a self-compatible (Sc) line of Brassica oleracea expressed a class II-like SLG (SLG-Sc) gene. Here, we report that the SLG-Sc glycoprotein is electrophoretically and immunochemically very similar to the recessive SLG-S15 glycoprotein, and is similarly expressed in stigmatic papillae. Moreover, by seed yield analysis, we observed that both alleles are associated with a self-compatibility response, in contrast with the other known recessive S haplotypes (S2 and S5). By genomic DNA blot analysis, we show the existence of molecular homologies between the Sc and S15 haplotypes, but demonstrate that they are not identical. On the other hand, we also report that the S2 haplotype expresses very low amounts of SLG glycoproteins, although it exhibits a self-incompatible phenotype. These results strongly question the precise role of the SLG gene in the molecular mechanisms that control the self-incompatibility reaction of Brassica.

Alleles↗

Expression of a self-incompatibility gene in a self-compatible line of Brassica oleracea.

In cruciferous plants, self-pollination is prevented by the action of genes situated at the self-incompatibility locus or S-locus. The self-incompatibility reaction is associated with expression of stigma glycoproteins encoded by the S-locus glycoprotein (SLG) gene. Only a few cases of self-compatible plants derived from self-incompatible lines in the crucifer Brassica have been reported. In these cases, self-compatibility was generally ascribed to the action of single genes unlinked to the S-locus. In contrast, we report here a line of Brassica oleracea var acephala with a self-compatible phenotype linked to the S-locus. By means of both biochemical and immunochemical analyses, we showed that this self-compatible (Sc) line nonetheless possesses stigmatic SLGs (SLG-Sc) that are expressed with a similar spatial and temporal pattern to that described for the SLGs of self-incompatible Brassica plants. Moreover, the SLG-Sc products segregate with the self-compatibility phenotype in F2 progeny, suggesting that changes at the S-locus may be responsible for the occurrence of the self-compatibility character. A cDNA clone encoding the SLG-Sc product was isolated, and the deduced amino acid sequence showed this glycoprotein to be highly homologous to the pollen recessive S2 allele glycoprotein. Hence, self-compatibility in this Brassica Sc line correlates with the expression of a pollen recessive-like S allele in the stigma.

Amino Acid Sequence↗

[Maxillofacial manifestations of Cowden's disease. Apropos of 2 cases].

Two new cases of Cowden's disease observed in two young sisters are reported. The diagnosis was based on the clinical observation of oral mucosa lesions (gingival hyperplasia, papillomatosis, scrotal tongue). In one case, visceral manifestations in childhood were observed. Cowden's disease is a familial affection (autosomal dominant trait) characterized by association of oral mucosa and skin lesions and of multiple hamartomas involving glandular tissues. Clinical and familial history investigations are justified by the high risk of malignant tumors (breast carcinoma).

Adolescent↗

Esophageal prothesis for neoplastic stenosis. A prognostic study of 77 cases.

Esophageal prothesis (EP) is a palliative treatment for inoperable neoplastic stenosis. Ninety-one patients were candidates for EP placement between 1978 and 1983. EP placement was successful in 77 patients. Of these: 85% had primary esophageal cancer; 15% had bronchial or mediastinal carcinoma; 83% had stenosis in the middle third. Dysphagia was the main symptom in 84%, and tracheoesophageal fistulae in 16%. The observed median survival was 3.2 months +/- 1.9 (2 standard deviations [SD]), and the survival rate at 1 year was 7%. With the single-factor analysis method, not one of 12 factors presented any significance (age, sex, general status, anterior treatment, lesion site, endoscopic feature, stenosis diameter, histologic features, metastasis, tracheal involvement, symptoms, and type of prothesis). Two factors had a P value approaching significance: there were slightly longer survival rates in patients with a stenosis diameter less than 7 mm (P less than 0.07), and with stenosis located in the lower third and cardia (P less than 0.07). By multivariate analysis (Cox model), prognostic significance was found in only one factor: the location in the lower third and cardia (P = 0.002, relative risk = 3). The quality of life after EP placement was briefly improved: 80% of 73 evaluable patients had improvement in dysphagia for a mean duration of 3.7 months +/- 2 (2 SD), especially patients with a good general status (0 and 1; P less than 0.01); and 45% of patients had improvement of their general status for a mean duration of 4.1 months +/- 2 (2 SD). Minor complications related to EP placement were observed in 40% of patients (pain, obstruction, and mobilization), and severe complications were observed in 20% (perforation, 11 cases; fistulae, 5 cases, with death in 3 cases; hemorrhages, 4 cases). In conclusion, EP is a good palliative treatment, although complications are frequent, and evaluation of patient comfort is required to compare this procedure with other available techniques.

Actuarial Analysis↗

Alterations of GABA-mediated synaptic transmission in human epilepsy.

Although animal models consistently indicate that gamma-amino-butyric acid (GABA) synaptic function (GABA levels, synthesis, uptake and/or receptors) is decreased in seizure states, there is little evidence to date in support of such a hypothesis for human epilepsy. This chapter presents the results of an in-depth study of the activity of the GABA-synthesizing enzyme L-glutamic acid decarboxylase (GAD) in brain tissue removed during neurosurgical resection for intractable epilepsy. The tissue studied is unique in that identified (by stereo EEG) foci were excised (rather than large blocks of tissue containing mixtures of foci and nonepileptic material) and compared with nonepileptic (stereo EEG and morphological definitions) tissue from the same patients. In patients in which there was no indication of a tumor, GAD activity in the foci was low in more than 50% of the patients examined. Furthermore, when the population distribution of GAD was compared in epileptic versus nonepileptic tissue fragments from all patients, the peak distribution of epileptic tissue fragments occurred at much lower GAD activities than for the nonepileptic fragments (0-20 versus 41-80 nmol CO2/mg protein X hr, respectively). A small subgroup of epileptic fragments occurred with a normal GAD distribution, indicating that the presence of an epileptic focus was not invariably associated with low GAD activity. When the low levels of GABA "A" binding sites in these epileptic tissue fragments are taken into consideration in combination with the low GAD levels, then it can be estimated that 60 to 70% of the present patient population had deficient GABAergic transmission in epileptic foci as compared to nonepileptic brain tissue from the same patients. It follows that the GABA hypothesis of human epilepsy is not an exclusive or unitary hypothesis, and some patients appear to have normally functioning GABA synapses (as assessed biochemically) in epileptogenic areas. Thus, other neurotransmitter and neurohumoral systems certainly play a role in the epileptic process.

Animals↗

Ultrastructural detection of fucosyl residues at the surface of axenically grown maize roots/sequential use of UeA lectin and fucosyl ferritin as the specific glycosylated marker.

This study reports the ultrastructural detection of fucosyl residues at the surface of axenically grown roots of Zea mays. The method used involved sequential binding of UeA lectin to the root and coupling of the bound UeA with fucosyl ferritin. Superficial dense ferritin labelling was found in the slime droplet ensheathing the root cap and in the external layer of the three-layered epidermal root surface. This pattern of binding of the fucose-specific lectin UeA suggests an overall distribution of the fucosyl residues on the root surface. Their localization within slime components of the root cap and the root epidermis is also assessed by the use of PATAg controls. Treatment of whole roots with alpha-L-fucosidase was ineffective in removing the fucosyl residues present at the root surface. The biological role of the fucosyl residues at the root-soil interface is discussed.

Cell Membrane↗