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Biomedical subjects

A M Rodriguez

Publications and source records attributed to A M Rodriguez.

At least 37 records · Page 2Linked to original sources
PubMed1997

Interferon-gamma rescues HLA class Ia cell surface expression in term villous trophoblast cells by inducing synthesis of TAP proteins.

Human placental trophoblast cells that constitute the materno-fetal interface during pregnancy escape maternal alloimmune attack. The different trophoblast cell subpopulations have developed efficient regulatory mechanisms to prevent expression of beta2-microglobulin-associated HLA class Ia molecules at their cell surface. We previously reported the presence of HLA class Ia messages in villous cytotrophoblast cells and in the syncytiotrophoblast differentiated in vitro purified from term placenta. In this study, we found that these transcripts are translated in heavy chain proteins that are endoglycosidase H sensitive and thus retained in the endoplasmic reticulum or cis-Golgi. Moreover, these class Ia heavy chains can be co-immunoprecipitated with the chaperone protein calnexin resident in the endoplasmic reticulum. When these trophoblast cells are treated with interferon (IFN)-gamma, part of the class Ia heavy chains become endoglycosidase H resistant, demonstrating that they have left the endoplasmic reticulum. Furthermore, after such a treatment, these heavy chains are detectable at the cell surface of these trophoblast cells, as assessed by two-color flow cytometry analysis and immunoprecipitation of cell surface biotinylated proteins using the W6/32 anti-HLA class I monoclonal antibody (mAb). IFN-gamma treatment induces a significant enhancement of the transcription of transporters associated with antigen processing (TAP1 and TAP2) rather than an increase of HLA class I or beta2-microglobulin messages. Finally, we demonstrate that an anti-TAP1 mAb co-immunoprecipitates TAP1 proteins and HLA class Ia heavy chains in these IFN-gamma-treated trophoblast cells. Thus, the constitutive absence of HLA class Ia cell surface expression in term villous cytotrophoblast and syncytiotrophoblast is likely to be due to a lack of transporter proteins that participate in the proper assembly of these molecules in the endoplasmic reticulum. Such a defect can be modified upon IFN-gamma treatment.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1997

Mandibular widening by intraoral distraction osteogenesis.

Transverse mandibular deficiency with crowding of the mandibular anterior teeth is frequently present in patients with Class I and II malocclusions. The hallmarks of treatment by compensating orthodontics, functional appliances or orthopaedic devices are instability, compromised periodontium and compromised facial aesthetics. A new surgical technique has been developed to widen the mandible. The method is based upon gradual osteodistraction following vertical interdental symphyseal osteotomy. Ten patients with transverse mandibular deficiency and significant dental crowding were treated by symphyseal distraction and subsequent non-extraction decompensating orthodontic treatment. Either an intraoral tooth-borne Hyrax appliance or a new custom-made bone-borne osteodistractor was used to gradually widen the mandible. The surgical procedures were accomplished under local anaesthesia and intravenous sedation in an ambulatory surgical setting using an individualized distraction protocol. The appliances were activated 7 days after symphyseal osteotomies, once each day at a rate of 1 mm per day and stabilized for 30-40 days after distraction. After the segments were distracted, non-extraction orthodontic alignment of the mandibular anterior teeth was accomplished. The symphyseal distraction gaps were bridged by new bony regenerate. Distraction osteogenesis provided an efficient surgical alternative to orthognathic surgery for widening the mandible and treatment of transverse mandibular deficiency without extraction of teeth.

Adolescent↗
PubMed1996

A gene similar to PKD1 maps to chromosome 4q22: a candidate gene for PKD2.

We report a partial cDNA sequence that encodes a protein, dubbed "polycystwin," with 21% identify and 46% similarity to amino acids 3688-4109 of the carboxyl terminus of polycystin, the gene product of the autosomal dominant polycystic kidney disease locus located on chromosome 16 at band p13 (PKD1). Northern analysis demonstrates that the R48321 gene is expressed in all tissues examined, including both adult and fetal kidneys. Finally, in situ hybridization studies localize this novel gene to 4q22, where PKD2, the second most common locus for ADPKD, is known to map. Therefore, R48321 is an excellent candidate gene for PKD2.

Amino Acid Sequence↗
PubMed1996

Topological studies of the membrane component of the OleC ABC transporter involved in oleandomycin resistance in Streptomyces antibioticus.

The OleC ABC transporter of Streptomyces antibioticus is constituted by an ATP-binding protein (OleC) and a hydrophobic protein (OleC5). Here we present experimental evidence demonstrating that the OleC5 protein is an integral membrane protein and we propose a topological model for its integration into the membrane. This model is based on the generation of hybrid proteins between different regions of OleC5 and a Escherichia coli beta-lactamase (BlaM) and the determination of the minimal inhibitory concentrations to ampicillin in these constructions. Fusions were generated both by cloning specific fragments of oleC5 and by creating ExoIII nested deletions of the gene. In the topological model proposed there will be six alpha-helix transmembrane regions, two cytoplasmic and four periplasmic loops and a hydrophobic linker domain.

ATP-Binding Cassette Transporters↗
PubMed1996

Effectiveness of a whole cell pertussis vaccine in child-care centers and schools.

BACKGROUND: Pertussis has substantially increased in Quebec, Canada, since 1990. We estimated pertussis vaccine effectiveness and vaccine coverage in child-care centers and elementary schools. METHODS: Two retrospective cohort studies were simultaneously conducted. One included 4482 children attending 88 public child-care centers and the other included 3429 pupils in 14 elementary schools. Cough and pertussis symptoms were assessed through a questionnaire and medical records; immunization status was ascertained by examination of written records. RESULTS: In child-care centers 95% of children had received at least three vaccine doses at the beginning of the follow-up; in schools more than 98% of pupils had received at least 4 doses. With > or = 4 doses of vaccine and a standard case definition used for surveillance (cough > or = 2 weeks, > or = 1 pertussis symptom and no other apparent cause for cough), vaccine effectiveness was estimated at 61% (95% confidence interval, 44 to 72%) in child-care centers and at 60% (95% confidence interval, 10 to 82%) in schools. With the same number of doses but a case definition requiring a cough > or = 5 weeks, vaccine effectiveness increased to 71% (95% confidence interval, 49 to 83) in child-care centers and to 86% (95% confidence interval, 66 to 94%) in schools. CONCLUSIONS: The increase in pertussis in Quebec is not caused by a low vaccine coverage. A low vaccine effectiveness may contribute to the resurgence of pertussis in the past decade.

Canada↗
PubMed1996

Placental expression of HLA class I genes.

This article presents an overview of the more recent data dealing with the constitutive, transcriptional, and translational expression of classical class Ia and nonclassical HLA-E and -G class Ib products in the different trophoblast cell subpopulations that constitute the maternofetal interface during human pregnancy. Of particular interest is the expression of alternatively spliced HLA-G transcriptional isoforms that may be translated in membrane-bound or soluble protein products. Molecular regulatory mechanisms that may control the differential expression of class Ia and class Ib molecules, according to the cell types, state of differentiation, and stages of gestation are also examined. They may operate at the levels of transcription, translation and/or transport of proteins to the cell surface. Functional significance of the absence of detectable cell surface expression of class Ia molecules in all trophoblast cell subpopulations, and of the presence of membrane-bound HLA-G products in extravillous cytotrophoblast cells is finally questioned.

Female↗
PubMed1996

Somatostatin antisense oligodeoxynucleotide-mediated stimulation of lymphocyte proliferation in culture.

We have previously shown that somatostatin (SRIF) is synthesized in B and T lymphocytes of rat spleen and thymus and released into the medium of cultured lymphocytes. To determine the role of SRIF in the control of lymphocytes proliferation, the expression of SRIF in normal lymphocytes was inhibited using a 3'-terminal phosphorothioate-modified antisense oligonucleotide complementary to a sequence that includes the translation start site of the rat SRIF messenger RNA. Spleens were obtained from adult male rats, and their lymphocytes were cultured for 24 or 72 h to measure SRIF content and cell proliferation, respectively. For the proliferation studies, [3H]thymidine was incorporated during the final 18 h. The lymphocytes were incubated with 15-30 micrograms/ml SRIF antisense and control antisense. SRIF antisense (25 micrograms/ml) increased lymphocyte proliferation 15-fold (P < or = 0.001), reaching a plateau (25- to 30-fold increase) between 25-30 micrograms/ml SRIF antisense. SRIF was extracted from lymphocytes and measured by RIA. Levels of SRIF content were almost undetectable with 10 micrograms/ml antisense and were significantly lower (P < or = 0.01) with 25 micrograms/ml antisense. When RC 160 (10(-5) M), a SRIF agonist analog, was used in the incubation, the stimulation of cell proliferation exerted by the SRIF antisense was completely abolished. Control antisense had no effect on proliferation or SRIF content. These findings indicate that 1) lymphocytes in culture are able to incorporate SRIF antisense; and 2) SRIF antisense inhibits the expression of lymphocytic SRIF, which leads to lymphocyte proliferation. In conclusion, cell proliferation is dramatically increased by eliminating the expression of SRIF from the lymphocytes, which indicates that in vitro SRIF is acting in a paracrine and/or autocrine fashion to inhibit lymphocyte proliferation.

Animals↗
PubMed1995

Detection of membrane-bound HLA-G translated products with a specific monoclonal antibody.

A monomorphic anti-HLA-G monoclonal antibody (mAb) was obtained by immunization of HLA-B27/human beta 2-microglobulin double-transgenic mice with transfected murine L cells expressing both HLA-G and human beta 2-microglobulin. This mAb, designated BFL.1, specifically recognizes, by flow cytometry analysis, the immunizing HLA-G-expressing cells, whereas it does not bind to parental untransfected or to HLA-B7- and HLA-A3-transfected L cells, suggesting that it distinguishes between classical HLA-A and -B and nonclassical HLA-G class I molecules. This was further assessed by the absence of BFL.1 reactivity with a number of human cell lines known to express classical HLA class I proteins. In addition, we showed that the BFL.1 mAb also labels HLA-G-naturally-expressing JEG-3 and HLA-G-transfected JAR human choriocarcinoma cell lines as well as a subpopulation of first-trimester placental cytotrophoblast cells. Further biochemical studies were performed by immunoprecipitation of biotinylated membrane lysates: BFL.1, like the monomorphic W6/32 mAb, immunoprecipitated a 39-kDa protein in HLA-G-expressing cell lines, a size corresponding to the predicted full-length HLA-G1 isoform. However, in contrast to W6/32, which immunoprecipitates both classical and nonclassical HLA class I heavy chains, BFL.1 mAb does not recognize the class Ia products. Such a mAb should be a useful tool for analysis of HLA-G protein expression in various normal and pathological human tissues and for determination of the function(s) of translated HLA-G products.

Animals↗
PubMed1995

Methylation status and transcriptional expression of the MHC class I loci in human trophoblast cells from term placenta.

Of the various molecular regulatory mechanisms that may be used by human trophoblast cells to down-regulate expression of HLA class I genes, we chose to investigate the methylation of DNA, generally associated with inhibition of transcription. We analyzed the methylation status of different HLA class I loci in villous and extravillous cytotrophoblast cells and in vitro-differentiated syncytiotrophoblast, purified from human term placenta, as well as in the human trophoblast-derived JAR and JEG-3 cell lines. We then compared methylation status and transcriptional activity. An inverse relationship was established between JAR and JEG-3: HLA-A, -B, and -G are methylated and repressed in JAR, whereas in JEG-3, HLA-A is methylated and repressed but HLA-B and -G are partially methylated and transcribed. HLA-E is unmethylated and transcribed in both cell lines. Apart from HLA-E, which is always unmethylated and transcribed, no such relationship exists for the other class I loci in trophoblast cells. Whereas nonclassical HLA-G and classical HLA-A and -B class I genes are undermethylated in both cytotrophoblast and syncytiotrophoblast, they are clearly transcribed in the former but minimally transcribed in the latter subpopulation. Thus, the down-regulation of class I gene expression in the in vitro-differentiated syncytiotrophoblast is unlikely to be caused by DNA methylation. Furthermore, there is no detectable expression of any class I molecule at the cell surface of either trophoblast cell subpopulation, suggesting a negative control on translation and/or on the secretory pathway to the plasma membrane.

Base Sequence↗
PubMed1995

A cytochrome P450-like gene possibly involved in oleandomycin biosynthesis by Streptomyces antibioticus.

A cosmid clone from an oleandomycin producer, Streptomyces antibioticus, contains a large open reading frame encoding a type I polyketide synthase subunit and an oleandomycin resistance gene (oleB). Sequencing of a 1.4-kb DNA fragment adjacent to oleB revealed the existence of an open reading frame (oleP) encoding a protein similar to several cytochrome P450 monooxygenases from different sources, including the products of the eryF and eryK genes from Saccharopolyspora erythraea that participate in erythromycin biosynthesis. The oleP gene was expressed in Escherichia coli as a fusion protein to a maltose-binding protein. Using polyclonal antibodies against this fusion protein it was observed that the synthesis of the cytochrome P450 was in parallel to that of oleandomycin. The cytochrome P450 encoded by the oleP gene could be responsible for the epoxidation of carbon 8 of the oleandomycin lactone ring.

Amino Acid Sequence↗
PubMed1994

Adhesive properties of choriocarcinoma cells toward lymphocytes activated or not by interleukin-2.

Choriocarcinoma cells (CC) in vitro are resistant to NK lysis but sensitive to lysis by blood or decidual effectors activated by interleukin-2 (IL-2). Because lytic activity requires a step of adhesion, the adhesive properties of the choriocarcinoma cells BeWo, JEG-3, and JAR were examined functionally toward peripheral blood lymphocytes. The adhesion of lymphocytes to choriocarcinoma cells was very low and did not increase after stimulating lymphocytes with IL-2. As demonstrated by cytofluorimetry analysis, choriocarcinoma cells and cytotrophoblast cells prepared from term placenta expressed intercellular adhesion molecule-1 (ICAM-1), whereas only CC expressed CD56. Tumor necrosis factor-alpha or interferon-gamma increased the expression of ICAM-1 on choriocarcinoma cells without modifying the adhesion of lymphocytes to choriocarcinoma cells. These results suggest that resistance of choriocarcinoma cells to lysis by cytotoxic effectors could partially be attributed to the low level of lymphocyte adhesion to these cells.

Cell Adhesion↗
PubMed1994

Cantilever and implant biomechanics: a review of the literature. Part 1.

In Part 1 of this two-part literature review, the biomechanics of cantilever fixed partial dentures are reviewed. Theoretical constructs of implant biomechanics with special emphasis on implant-supported cantilevers are also discussed. Finally, an overview of the literature regarding occlusal forces generated by patients with implant-supported prostheses is presented.

Biomechanical Phenomena↗
PubMed1994

Cantilever and implant biomechanics: a review of the literature, Part 2.

In Part 2 of this literature review, a summary of the literature regarding the determination of acceptable cantilever lengths for fixed implant prostheses is presented. Studies examining the possible effects of biomechanical stress on both the implant prosthesis and the supporting bone are also discussed.

Dental Implants↗
PubMed1994

Structure-activity relationship in the cytoprotective effect of helenalin and related compounds.

Our previous reports indicate that the cytoprotective effect of helenalin and several sesquiterpene lactones is mediated through a Michael reaction between the sulfhydryl containing peptides of the gastric mucosa and Michael acceptors present in the sesquiterpene lactone molecules. In the present work the different alternative active sites in molecules containing up to three places which could act as viable Michael acceptors are evaluated. To do this, an extensive conformational and electronic study of helenalin and its derivatives was carried out. The experimental and theoretical results obtained show the gamma-lactone and cyclopentenone groups to be the biologically active places of the molecules under study, whereas tiglic and angelic substituents in these compounds show different behavior as Michael acceptors.

Cardiotonic Agents↗
PubMed1993

Evaluation of strain at the terminal abutment site of a fixed mandibular implant prosthesis during cantilever loading.

PURPOSE: Cantilever lengths from 10 mm to 20 mm have been empirically recommended for Brånemark fixed mandibular implant prostheses. However, functional stresses generated within the framework and at the crestal bone associated with various cantilever lengths have not been well researched. The purpose of this investigation was to evaluate the strain generated within an implant-supported prosthesis and on a simulated bone surface during functional cantilever loading. MATERIALS AND METHODS: A symmetrical mandibular fixed-implant framework supported by six Nobelpharma 7.0 x 4.0-mm abutments and 15.0 x 4.0-mm fixtures was fabricated. The fixtures were embedded in a simulated bone matrix of polymethyl methacrylate resin. Fourteen different arrangements of active supporting abutments were tested during 15 lb unilateral static cantilever loading 7 mm, 14 mm and 20 mm distal to the terminal abutments. T-rosette strain gauges were placed immediately distal to the terminal abutment site on the right side of the framework and on the corresponding simulated bone surface. RESULTS: There was no difference in framework microstrain as abutment number and arrangement were varied. Microstrain distal to the terminal abutment increased significantly with increasing cantilever length. Distal abutment microstrain increased 213% (63 mu epsilon to 197 mu epsilon) when cantilever length was increased from 7 mm to 14 mm and an additional 55% (197 mu epsilon to 306 mu epsilon) when cantilever length was increased from 14 mm to 20 mm. Overall, microstrain increased 306% when cantilever length was increased from 7 mm to 20 mm. Microstrain on the framework was always tensile (positive). Microstrain at the simulated bone reached higher maximum levels than on the framework (-588 mu epsilon versus 314 mu epsilon) and was compressive in nature (negative). In contrast to framework microstrain, microstrain at the simulated bone site varied dramatically with changes in abutment arrangement. Strains observed at the simulated bone surface increased dramatically as the distance to the adjacent active abutment increased or as the anterior-posterior span of abutments decreased. Distal abutment microstrain also increased significantly at the bone site as cantilever length increased, however, percent increases were less (7 mm to 14 mm, 55%; 14 mm to 20 mm, 30%; 7 mm to 20 mm, 101%). CONCLUSIONS: The results of this study indicate that an optimum biomechanical environment should exist when cantilever spans exceeding 7 mm are planned regardless of the number of supporting abutments. Strain transmitted to the crestal bone can be decreased by maximizing the number and anterior-posterior spread of supporting fixtures while minimizing the distance between the distal abutment and its adjacent abutment.

Alveolar Process↗
PubMed1993

Differences between human sperm and somatic cell DNA in CpG methylation within the HLA class I chromosomal region.

PROBLEM: We investigated the possible negative regulatory mechanisms that repress classical human leukocyte antigen (HLA) class I gene expression in human spermatozoa and searched for novel testis-specific coding sequences that might be present in MHC class I chromosomal region. METHOD: We performed a comparative DNA methylation analysis of this genomic region in both purified human spermatozoa and mononuclear blood cells from the same donors, using methylation-sensitive restriction enzymes followed by classical or pulsed field gel electrophoresis and hybridization with HLA class I locus-specific probes. RESULTS: Unmethylated CpG sites were detected in the 3' part of HpaII tiny fragments of the HLA-F and HLA-G genes in spermatozoal DNA. In contrast, no difference was observed in the methylation status of the HLA-B, HLA-C, and HLA-E genes between germ and somatic cells. CpG unmethylation events were also detected in several parts of this chromosomal region (outside the known loci) in spermatozoal DNA. CONCLUSIONS: These results suggest that this genomic region undergoes changes in its DNA methylation pattern during the developmental process. We hypothesize that these dynamic changes have functional importance, including a possible transcriptional activity of nonclassical class I genes and/or as yet undescribed testis-specific coding sequences.

Base Sequence↗