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Biomedical subjects

M Richard

Publications and source records attributed to M Richard.

At least 73 records · Page 4Linked to original sources

Allele frequency data for VNTR locus D17S79: identification of an internal HaeIII polymorphism in the black population.

D17S79 is one of several highly polymorphic VNTR loci commonly used to generate DNA profiles for forensic identity testing and parentage determination. In this study, we used Southern hybridization analysis of HaeIII-digested genomic DNA to compile D17S79 allele frequency databases for the Caucasian, Southeast Asian, East Indian, and Black populations of Southern Ontario. In our Black population sample, low-molecular-weight D17S79 alleles are relatively common and approximately 7% of the individuals have three-band hybridization patterns due to a HaeIII restriction site polymorphism within a common D17S79 allele. The internal HaeIII site generates an allele consisting of two HaeIII fragments of approximately 980 and 420 bp. The estimated frequency of this "split-allele" in our Black population is 0.035. Since this allele was not observed in the other population groups, the internal HaeIII polymorphism probably originated within the Black population. Although the existence of low-molecular-weight alleles and the internal HaeIII polymorphism could affect the interpretation of some VNTR patterns, they are of little or no forensic consequence to the frequencies assigned to VNTR profiles.

Alleles↗

Chloroplastic genomes of Ginkgo biloba and Chlamydomonas moewusii contain a chlB gene encoding one subunit of a light-independent protochlorophyllide reductase.

We have cloned and sequenced a Chlamydomonas moewusii chloroplastic DNA fragment that includes a 563 amino-acid open reading frame (ORF563, chlB) presenting 89% amino-acid homology with ORF513 from Marchantia polymorpha. It is also homologous to ORF510 from Pinus thunbergii but includes two insertions absent in both M. polymorphia and P. thunbergii. The derived polypeptide is 54% similar to the product of bchB from Rhodobacter capsulatus, identified as one subunit of a light-independent NADH-protochlorophyllide reductase. We also isolated and sequenced an homologous chloroplastic gene from the gymnosperm Ginkgo biloba. Northern hybridizations performed on RNA isolated from synchronized Chlamydomonas eugametos cells showed higher expression between the tenth hour of light and the eighth hour of darkness, peaking during the first 2 h of darkness.

Amino Acid Sequence↗

Integration of a vector containing a repetitive LINE-1 element in the human genome.

Mammalian cells contain numerous nonallelic repeated sequences, such as multicopy genes, gene families, and repeated elements. One common feature of nonallelic repeated sequences is that they are homeologous (not perfectly identical). Our laboratory has been studying recombination between homeologous sequences by using LINE-1 (L1) elements as substrates. We showed previously that an exogenous L1 element could readily acquire endogenous L1 sequences by nonreciprocal homologous recombination. In the study presented here, we have investigated the propensity of exogenous L1 elements to be involved in a reciprocal process, namely, crossing-overs. This would result in the integration of the exogenous L1 element into an endogenous L1 element. Of over 400 distinct integration events analyzed, only 2% involved homologous recombination between exogenous and endogenous L1 elements. These homologous recombination events were imprecise, with the integrated vector being flanked by one homologous and one illegitimate junction. This type of structure is not consistent with classical crossing-overs that would result in two homologous junctions but rather is consistent with one-sided homologous recombination followed by illegitimate integration. Contrary to what has been found for reciprocal homologous integration, the degree of homology between the exogenous and endogenous L1 elements did not seem to play an important role in the choice of recombination partners. These results suggest that although exogenous and endogenous L1 elements are capable of homologous recombination, this seldom leads to crossing-overs. This observation could have implications for the stability of mammalian genomes.

Cell Line↗

[Spas in Gallo-roman Aquitaine].

We have various witnesses about spas during the gallo-roman period; it is in Luchon, Dax, Bagnères-de-Bigorre and Amélie les Bains, that we have the most important number of documents, coins, various chinas, and votive stones. It is interesting to note that after two thousand years the therapeutic qualities of mineral hot waters springs are still recognized; and in addition to the four mentioned spas, we can add: Barbotan, Prechacq, Saint-Christau, Capvern, Encausse, Salies-du-Salat, Ax-les-Thermes, Aulus, Alet, Rennes-les-Bains. It is interesting also to note that goddesses exhibited near the springs, were later replaced by Saints, which were the only explanation about the activity of the springs during several centuries.

Balneology↗

[Serum phospholipase A2 activity in osteoarthritis].

Serum phospholipase A2 activity in 67 osteoarthritis patients and 17 controls was determined using a radiolabeled specific substrate. Serum phospholipase A2 activity was significantly higher in osteoarthritis patients (115 +/- 73.6 dpm/h/ml) than in controls (45 +/- 25 dpm/h/ml) (p = 0.002). In 41 osteoarthritis patients, serum phospholipase A2 activity was unrelated to age, time since onset of osteoarthritis symptoms, duration of morning stiffness, Lequesne's index, roentgenographic stage of osteoarthritis, number of joints with osteoarthritis, erythrocyte sedimentation rate, or serum C-reactive protein levels. In 12 osteoarthritis patients who were evaluated twice at a mean interval of 46 days, changes in serum phospholipase A2 activity were unrelated to changes in Lequesne's index. Blind evaluation of long-term joint space loss was performed in 14 patients; serum phospholipase A2 activity increased only in those patients with progressive joint space loss, but the difference was not statistically significant as compared with the controls. These data suggest that serum phospholipase A2 activity is not useful in practice as a marker for osteoarthritis.

Adult↗

Targeting of histone tails by poly(ADP-ribose).

After Zn2+ finger-mediated binding to a DNA break, poly(ADP-ribose) polymerase becomes automodified with long polymers of ADP-ribose. These nucleic acid-like polymers may facilitate DNA repair by noncovalently interacting with neighboring proteins. Using a novel screening technique, we have identified histones as the predominant poly(ADP-ribose)-binding species in human keratinocytes, rat hepatocytes, frog eggs, and yeast. Polymer binding is confined specifically to the histone domains responsible for DNA condensation, i.e. histone tails. Our results indicate that polymers of ADP-ribose are targeted to sites of DNA strand breaks by poly(ADP-ribose) polymerase and subsequently function to alter chromatin conformation through noncovalent interactions with histone tails.

Animals↗

Shedding of GD2 ganglioside in patients with retinoblastoma.

Retinoblastoma is a rare tumor of the young child with an intraocular localization that leads to certain problems of diagnosis. With the aim of defining a biochemical marker--which is still lacking for this disease--the gangliosides of a pool of fresh retinoblastoma tumors were analyzed. The ganglioside pattern was shown to have GM3, GM2, GM1, GD3, GD2, GD1b and GT1b as the major components. The occurrence of a high concentration of GD2 in the tumors led us to investigate the possibility of changes in the level of GD2 in the sera of retinoblastoma patients, using quantitative immunostaining with GD2-specific mouse monoclonal antibodies (MAbs). In 9 out of 10 tumor-bearing patients, the serum level of GD2 ganglioside was significantly higher than the average value found in normal individuals. A 2-year follow-up of patients showed that successful treatment resulted in a rapid decrease in the serum level of GD2 down to the normal range, from which a subsequent elevation was seen only in relapsing patients. Although the clinical study needs further development, the results obtained to date suggest that GD2 is shed in the serum of tumor-bearing patients and that the level of GD2 could be a potential serum marker of human retinoblastoma.

Child, Preschool↗

The effects of high molecular weight- and low molecular weight-heparins on superoxide ion production and degranulation by human polymorphonuclear leukocytes.

High molecular weight (HMW) and low molecular weight (LMW) heparins affected superoxide ion production and degranulation by polymorphonuclear leukocytes (PMNL) isolated from either chronic hemodialyzed patients or healthy controls. Low concentrations in HMW-heparin, below 1.76 aXa IU/ml for patients and 1.34 aXa IU/ml for controls, increased O2- production started by phorbol myristate acetate. High concentrations above these values decreased it. Increasing LMW-heparin concentrations constantly decreased O2- production using the same stimulus. Myeloperoxidase (MPO) released by PMNL was found to be significantly HMW- and LMW-heparins dose-dependent. The addition of calcium chloride significantly increased MPO release. Lactoferrin release was not dose-dependent of HMW- or LMW-heparins. However, an increase of the percentage of positive responses for lactoferrin release was observed in the simultaneous presence of HMW-heparin and CaCl2 compared to HMW-heparin alone. Lysozyme release was also not dose-dependent of HMW- or LMW-heparins. An increase of the percentage of positive responses for lysozyme release was observed in the presence of CaCl2 alone compared to HMW-heparin.

Adult↗

Characterization of gangliosides from normal and osteoarthritic human articular cartilage.

OBJECTIVE: Glycosphingolipids (GSLs) are biologically active molecules in the physiology and pathology of cells. Since changes in GSLs might be associated with the impaired metabolism of articular cartilage in osteoarthritis (OA), we investigated gangliosides from normal and OA human cartilage. METHODS: OA and control cartilage was obtained from patients with hip OA and femoral neck fracture, respectively. Gangliosides were extracted and quantified by determining their lipid-bound sialic acid concentration. Major gangliosides were identified by immuno-detection on thin-layer plates, purified by high performance liquid chromatography, and analyzed for their carbohydrate, fatty acid, and long-chain base composition. RESULTS: The total ganglioside content of OA cartilage was decreased by 40% (per mg of DNA). Major gangliosides, GM3 and GD3, separated into 3 on thin-layer chromatography bands. All were decreased except for the lowest migrating band of GM3, which was increased 5-fold. This ganglioside had the same carbohydrate moiety and fatty acids as the other two, but differed by a long-chain base composed mainly of C20-sphingosine. CONCLUSION: OA cartilage is characterized by a decrease in all gangliosides except GM3, which demonstrates a large increase in the lowest band. These results indicate that there are changes in the biochemical composition of chondrocyte membranes in OA. The causes and roles of these changes remain to be determined.

Aged↗

Metalloprotease activity, phospholipase A2 activity and cytokine concentration in osteoarthritis synovial fluids.

Collagenase, stromelysin and phospholipase A2 (PLA2) activity as well as interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF alpha) and interleukin 6 (IL-6) concentration were determined in the knee joint synovial fluid (SF) of 26 patients with osteoarthritis (OA) and with rheumatoid arthritis (RA). Collagenase and stromelysin were detected in 80.7 and 69.2% of OA SF, respectively. When present, the mean activity of both enzymes was approximately two times lower in OA than in RA SF. PLA2 activity was present in all SF with no significant difference between OA and RA SF. IL-1 beta, TNF alpha and IL-6 were found in 0, 96.1 and 84.6% of OA SF, respectively. Mean TNF alpha and IL-6 concentration was also lower in OA than in RA SF. Metalloprotease activity correlated weakly with IL-6 level and enzymatic activities were unrelated with TNF alpha in OA SF.

Adult↗

[Serum hyaluronic acid in osteoarthritis].

In this prospective study, serum hyaluronate (SH) was assayed using a radiometric method (Pharmacia) in 73 osteoarthritis patients and 39 controls. All assays were performed between 8 h 00 and 9 h 00 a.m. because SH levels exhibit circadian variations. SH levels were significantly higher in patients with osteoarthritis than in controls (92 +/- 66 micrograms/l and 39 +/- 21 micrograms/l, respectively, p = 0.0001). Among 50 patients with osteoarthritis, including 29 with knee involvement and 21 with hip involvement, SH levels were not correlated with morning stiffness, duration of symptoms, Lequesne's algofunctional index, erythrocyte sedimentation rate, C-reactive protein, severity of roentgenographic changes in the affected knee or hip, disease extension, or severity. The lack of any relationship between changes in SH levels and Lequesne's is index values in 25 patients or between SH levels and joint space narrowing evaluated retrospectively in 16 patients, as well as the prompt return to high SH levels after arthroplasty and synovectomy in 14 patients with hip joint osteoarthritis, suggest that this potential marker is not useful for monitoring osteoarthritis in a single joint.

Adult↗

Inhibition of metastatic behavior of murine osteosarcoma by hypophysectomy.

BACKGROUND: We recently reported that human osteogenic sarcoma cells are mitogenically responsive in tissue culture to insulin-like growth factor I (IGF-I), a mitogen important in the regulation of cellular proliferation of many tissues, including bone. PURPOSE: The present study was designed to determine whether these in vitro observations could be extended to an in vivo experimental system and whether reduction of IGF-I levels by hypophysectomy could inhibit the aggressive metastatic behavior of osteosarcoma. METHODS: We used standard competitive binding and affinity-labeling techniques to characterize the IGF-I-binding sites of MGH-OGS, a model of human osteosarcoma. Radioimmunoassay of serum, preprocessed to remove IGF-binding proteins, was used to quantitate IGF-I levels. In vitro proliferative response of MGH-OGS cells to IGF-I and other pituitary-dependent factors was determined by thymidine-incorporation experiments. In vivo growth of the neoplasm in 12 hypophysectomized C3H mice and in 14 control C3H mice was determined by serial measurements of implanted tumors and by gross and microscopic examination of the lungs for metastases. RESULTS: MGH-OGS exhibited specific binding sites for 1.39 pmol IGF-I per milligram MGH-OGS cellular membrane protein, a concentration similar to that which we previously reported for human osteosarcoma. In tissue culture, MGH-OGS exhibited mitogenic response to IGF-I (P less than .01) but not to other pituitary-dependent factors. Hypophysectomy reduced levels of circulating IGF-I to 15% of control, significantly inhibited local growth of MGH-OGS tumors (increased time for growth to 1 cm3 from 49 to 84 days, P less than .001), and profoundly inhibited metastatic behavior (decrease in mean number of metastases per host from 16 to less than one; P less than .001). CONCLUSIONS: This study is the first to document the profound inhibitory effect of hypophysectomy on the metastatic behavior of an experimental sarcoma. We conclude that the metastatic behavior exhibited by MGH-OGS osteosarcoma is dependent on pituitary factors, and we suggest that the inhibitory effects of hypophysectomy are related, at least in part, to the reduction of IGF-I levels.

Animals↗

Evaluation of bepridil efficacy by electrophysiologic testing in patients with recurrent ventricular tachycardia: comparison of two regimens.

The purpose of the study was to evaluate this effect of different doses of intravenous and oral bepridil on the induction of ventricular tachycardia. Thirty-eight patients underwent electrophysiologic evaluation for recurrent ventricular tachycardia (VT). Sustained monomorphic VT was induced by programmed ventricular stimulation, using up to three extrastimuli in all patients. The effects of intravenous bepridil (2 mg/kg) were evaluated during the initial study. Intravenous bepridil prevented the induction of sustained VT in eight patients (21%). Electrophysiologic study was repeated after oral bepridil. In six patients the study was stopped because of adverse effects or VT recurrence. Thirty-two patients underwent repeat study 7 days later, taking oral bepridil, 500 mg/day (n = 16) or 900/day (n = 16). A dose of 500 mg/day of bepridil prevented the induction of sustained VT in only one patient. A dose of 900 mg/day of bepridil prevented the induction of sustained VT in eight patients. There were no significant clinical adverse effects, except in one patient receiving intravenous bepridil. The response to intravenous bepridil did not predict the response to oral bepridil. The response to intravenous or oral bepridil was not related to the plasma level of bepridil but was related to a higher left ventricular ejection fraction. Eight patients (21%) in whom VTs were noninducible on oral bepridil were discharged on 300 mg/day of bepridil if their initial loading dose was 500 mg/day or on 600 mg/day if their initial loading dose was 900 mg/day. They remained free of VT during a follow-up of at least 6 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

Severe hypersensitivity reaction to injectable Gallium 67 in a worker exposed to silica.

Gallium 67 scintigraphy is employed in the evaluation of workers with possible pneumoconiosis. To our knowledge, however, a severe hypersensitivity reaction following the intravenous injection of Gallium 67 has not been described. We report the case of a worker chronically exposed to silica who developed an allergic cutaneous and severe articular reaction following the injection of Gallium 67 while being investigated for possible silicosis. Hilar adenopathy was noted on the chest roentgenogram and, retrospectively, circulating immune complexes were found in the patient's serum. The presence of a positive prick skin test to benzyl alcohol suggests that this preservative caused the hypersensitivity reaction.

Adult↗

Effect of non-steroidal anti-inflammatory drugs (NSAIDS) on glycosyltransferase activity from human osteoarthritic cartilage.

The effect of non-steroidal anti-inflammatory drugs (NSAIDs) on the activity of glycosyltransferases required for the synthesis of the polysaccharide chains of proteoglycans, was studied in human osteoarthritic cartilage in vitro. Using exogenous acceptors, salicylate and indomethacin suppressed the activity of glucuronyl- and xylosyltransferases in a concentration-dependent manner, but had little effect on N-acetylgalactosaminyl- and galactosyltransferases. When used at a concentration derived from the values found in the synovial fluid, salicylate, indomethacin and chloroquine significantly suppressed the activity of glucuronyl- and xylosyltransferases, while tiaprofenic acid, paracetamol (acetaminophen), floctafenine, ketoprofen, ibuprofen and tenoxicam had no effect on the enzymes. An alteration of some glycosyltransferases could explain the reported suppressive effect of some NSAIDs on cartilage proteoglycan synthesis.

Aged↗

Return of ATP/PCr and EEG after 75 min of global brain ischemia.

Acute, progressive global brain ischemia was induced in awake or anesthetized rats for 5-75 min. Ischemia was achieved with a subclavian-carotid artery occlusion technique (SCOT). After thoracotomy, both subclavian arteries (proximal to their vertebral branches) were tied-off and carotid artery catheter-snares installed. Results show progressive morphological, physiological and neurochemical damage when CBF was reduced from preischemic levels of 115 ml to 0 blood flow. 31P magnetic resonance spectroscopy of high energy phosphate metabolites in vivo showed loss of PCr and beta-ATP signals after 6 min brain ischemia. Energy metabolite levels, EEG and CBF normalized within hours after reperfusion. Degree of neuropathologic damage to hippocampal region appeared linearly related to the ischemic duration of ischemia. Thus, acute global brain ischemia resulted in loss of high energy phosphate metabolites, EEG and neuronal integrity in the hippocampal subfields. Reperfusion following short (5 min) or long (75 min) periods of global brain ischemia induced return of 31P-spectra, EEG and CBF to normal but was unable to reverse all of the neuronal damage at the end of the 72-h observation period.

Adenosine Triphosphate↗