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P Bailly

Publications and source records attributed to P Bailly.

At least 37 records · Page 2Linked to original sources

Time-course expression of polypeptides carrying blood group antigens during human erythroid differentiation.

The time course expression of blood group antigens was examined by flow cytometry using a two-phase liquid culture system that supports the proliferation and maturation of human erythroid progenitors from adult peripheral blood. The progression towards erythroid differentiation was followed by the expression changes of the transferrin receptor (CD71++) and glycophorin A (GPA+). Four main categories of blood group markers were identified: (i) those characterized by an early expression like ABO (A), Kell (K:2) and Rh50 which were detected in the Epo-independent phase 1, (ii) those including GPC (Gerbich, Ge antigens) and Fy6 which were expressed in the late phase 1, (iii) GPA (MN antigens), Wrb (Band 3/GPA interaction), Rh(D, Cc/Ee) and LW which appeared during the Epo-dependent phase 2 and (iv) those like Jk3 and Lub which were expressed in late phase 2. Regarding blood group molecules exhibiting adhesive properties (LW/ICAM-4, Oka and Lu) the most significant event was a sharp decrease of Oka (neurothelin) expression with the concomitant loss of ICAMs expression during the later stage of differentiation. These studies suggest that Oka, ICAMs and LW might contribute to the adhesive interactions involved in the formation of erythroblastic islands and attachment to stroma cells and the extracellular matrix. We also noted an asynchronous expression of the proteins that compose the core of the Rh complex, since Rh50 glycoprotein was expressed earlier than Rh(D, CE) proteins.

Antigens, CD↗

Extracellular K+ dependence of inward rectification kinetics in human left ventricular cardiomyocytes.

BACKGROUND: In human ventricular cells, the inwardly rectifying K+ current (IK1) is very similar to that of other mammalian species, but detailed knowledge about the K+-dependent distribution of open and blocked states during rectification and about the K+-dependent modulation of inactivation on hyperpolarization is currently lacking. METHODS AND RESULTS: We used the whole-cell patch-clamp technique to record IK1 in myocytes isolated from subendocardial layers of left ventricular septum from patients with nonfailing hearts with aortic stenosis and cardiac hypertrophy who were undergoing open-heart surgery. Outward currents were very small at voltages positive to the reversal potential but increased at high external [K+]. Chord conductance measurements and kinetic analyses allowed us to estimate the proportion of channels in the open state and of those showing either slow unblock or instantaneous unblock (the so-called slow or instantaneous "activation") on hyperpolarization: the distribution in the individual states was dependent on external [K+]. The proportion of channels unblocking slowly was greater than that of channels unblocking instantaneously on hyperpolarization from the plateau voltage range. Hence, because of the previously reported link between the presence of highly protonated blocking molecules and slow unblock kinetics, it is suggested that high cellular concentrations of spermine may account for the low outward current density recorded in these cells. The current decrease observed on extended hyperpolarization was significantly relieved by an increase in external [K+]. CONCLUSIONS: The pattern of IK1 current alterations observed in the present model of human ventricular hypertrophy might favor enhanced excitability and underlie ventricular arrhythmias, possibly via increased intracellular polyamine levels.

Action Potentials↗

Characterization of a UDP-Gal:Galbeta1-3GalNAc alpha1, 4-galactosyltransferase activity in a Mamestra brassicae cell line.

The binding of Bandeiraea simplicifolia lectin-I isolectin B4 on the endogenous glycoproteins of different insect cell lines led us to characterize for the first time a UDP-Gal:Galbeta1-3GalNAc alpha1, 4-galactosyltransferase in a Mamestra brassicae cell line (Mb). The study of the acceptor specificity indicated that the Mb alpha-galactosyltransferase prefers Galbeta1-3-R as acceptor, and among such glycans, the relative substrate activity Vmax/Km was equal to 20 microliters.mg-1.h-1 for Galbetal-3GlcNAcbeta1-O-octyl and to 330 microliters.mg-1.h-1 for Galbeta1-3GalNAcalpha-1-O-benzyl, showing clearly that Galbeta1-3GalNAc disaccharide was the more suitable acceptor substrate for Mb alpha-galactosyltransferase activity. Nuclear magnetic resonance and mass spectrometry data allowed us to establish that the Mb alpha-galactosyltransferase synthesizes one unique product, Galalpha1-4Galbeta1-3GalNAcalpha1-O-benzyl. The Galbeta1-3GalNAc disaccharide is usually present on O-glycosylation sites of numerous asialoglycoproteins and at the nonreducing end of some glycolipids. We observed that Mb alpha1,4-galactosyltransferase catalyzed the transfer of galactose onto both natural acceptors. Finally, we demonstrated that the trisaccharide Galalpha1-4Galbeta1-3GalNAcalpha1-O-benzyl was able to inhibit anti-PK monoclonal antibody-mediated hemagglutination of human blood group PK1 and PK2 erythrocytes.

Animals↗

Characterization of the gene encoding the human Kidd blood group/urea transporter protein. Evidence for splice site mutations in Jknull individuals.

The Kidd (JK) blood group is carried by an integral membrane glycoprotein which transports urea through the red cell membrane and is also present on endothelial cells of the vasa recta in the kidney. The exon-intron structure of the human blood group Kidd/urea transporter gene has been determined. It is organized into 11 exons distributed over 30 kilobase pairs. The mature protein is encoded by exons 4-11. The transcription initiation site was identified by 5'-rapid amplification of cDNA ends-polymerase chain reaction at 335 base pairs upstream of the translation start point located in exon 4. The 5'-flanking region, from nucleotide -837 to -336, contains TATA and inverted CAAT boxes as well as GATA-1/SP1 erythroid-specific cis-acting regulatory elements. Analysis of the 3'-untranslated region reveals that the two equally abundant erythroid transcripts of 4.4 and 2.0 kilobase pairs arise from usage of different alternative polyadenylation signals. No obvious abnormality of the Kidd/urea transporter gene, including the 5'- and 3'-untranslated regions, has been detected by Southern blot analysis of the blood of two unrelated Jknull individuals (B.S. and L.P.), which lacks all Jk antigens and Jk proteins on red cells, but was genotyped as homozygous for a "silent" Jkb allele. Further analysis indicated that different splice site mutations occurred in each variant. The first mutation affected the invariant G residue of the 3'-acceptor splice site of intron 5 (variant B.S.), while the second mutation affected the invariant G residue of the 5'-donor splice site of intron 7 (variant L.P.). These mutations caused the skipping of exon 6 and 7, respectively, as seen by sequence analysis of the Jk transcripts present in reticulocytes. Expression studies in Xenopus oocytes demonstrated that the truncated proteins encoded by the spliced transcripts did not mediate a facilitated urea transport compared with the wild type Kidd/urea transporter protein and were not expressed on the oocyte's plasma membrane. These findings provide a rational explanation for the lack of Kidd/urea transporter protein and defect in urea transport of Jknull cells.

Base Sequence↗

Flexible posterior mitral annuloplasty: five-year clinical and Doppler echocardiographic results.

BACKGROUND: Stabilization of the posterior annulus seems to be a critical factor to achieve a stable mitral valve repair. To assess the benefit of softer mural annuloplasty, we analyzed results obtained with the flexible linear reducer. METHODS: From 1985 to 1993, 120 patients, with pure mitral regurgitation, mainly degenerative, had a mitral reconstruction. Mean age was 64+/-11 years and 74% of the patients were in New York Heart Association functional class III or IV. RESULTS: Hospital mortality was 3.3%. Mean follow-up was 56+/-24 months. There were 23 late deaths; 10 valve-related including 7 sudden deaths. Two patients (1.7%) required a reoperation. Doppler echocardiographic studies revealed excellent valve function; 5-year freedom from significant regurgitation was 85.8%+/-5.4%. Mean mitral valve area was 2.76+/-0.77 cm2. Although 105 patients were in class I or II, 23 patients were not functionally improved. Previous myocardial infarction and shorter deceleration time of early filling were risk factors for worsening functional disability. CONCLUSIONS: This support provides stable repair with excellent clinical and echographic results. Previous myocardial infarction and noncompliant left ventricle negatively influence outcome.

Adult↗

Cultured neonatal rat cardiomyocytes and 99mTc-sestamibi to assess the direct effects of cardioplegia.

The efficacy of cardioplegia in neonatal myocardial protection is still a matter of debate. 99mTc-sestamibi cellular accumulation reflects sarcolemmal and mitochondrial electrical gradients. It was used to monitor the direct effects of two cardioplegic solutions, modified St Thomas' Hospital and Bretschneider, on normoxic and metabolically-inhibited cultured cells. Cellular accumulation of 99mTc-sestamibi, expressed by the ratio between intra and extra cellular concentrations, was assessed in three different sets of neonatal rat cardiomyocytes. Cells were either treated with different concentrations of modified St Thomas' solution (50, 75, 100%), they were treated or recovering from a treatment with modified St Thomas and Bretschneider solutions at 50% concentrations, or were recovering from treatment with modified St Thomas' and Bretschneider solution at 50% concentrations mixed with metabolic inhibitors. Cardioplegia depressed the tracer accumulation in a concentration-dependent manner. This effect was independent of the type of cardioplegia (120-min uptake, as a percentage of control values, modified St Thomas' 68+/-12 and Bretschneider 59+/-7) and was rapidly reversible. Cardioplegia was unable to prevent the depression of tracer accumulation induced by metabolic inhibitors and even induced a deleterious effect (120-min uptake, as a percentage of control values, metabolic inhibitors 69+/-12, metabolic inhibitors + modified St Thomas 38+/-14, metabolic inhibitors + Bretschneider 43+/-6) during recovery after 30 min of metabolic inhibition. It was concluded that cardioplegia has an apparent detrimental effect on neonatal cardiomyocytes accumulation of 99mTc-sestamibi during recovery from an ischaemic-like insult.

Animals↗

Insights into the structure and function of membrane polypeptides carrying blood group antigens.

In recent years, advances in biochemistry and molecular genetics have contributed to establishing the structure of the genes and proteins from most of the 23 blood group systems presently known. Current investigations are focusing on genetic polymorphism analysis, tissue-specific expression, biological properties and structure-function relationships. On the basis of this information, the blood group antigens were tentatively classified into five functional categories: (i) transporters and channels, (ii) receptors for exogenous ligands, viruses, bacteria and parasites, (iii) adhesion molecules, (iv) enzymes and, (v) structural proteins. This review will focus on selected blood groups systems (RH, JK, FY, LU, LW, KEL and XK) which are representative of these classes of molecules, in order to illustrate how these studies may bring new information on common and variant phenotypes and for understanding both the mechanisms of tissue specific expression and the potential function of these antigens, particularly those expressed in nonerythroid lineage.

Anemia, Hemolytic, Congenital↗

Risks and routes for ventilator-associated pneumonia with Pseudomonas aeruginosa.

In a prospective study, we screened specimens from 190 mechanically ventilated patients hospitalized in a surgical intensive care unit, and from the environment to assess risks and routes of colonization/infection. Specimens from various sites were collected on admission and once a week throughout each patient's stay. All P. aeruginosa isolates were typed by determination of DNA patterns. Data were collected from patients to identify risk factors. In vitro production of exoenzymes of different strains were compared. Forty-four patients were colonized with P. aeruginosa on the bronchopulmonary tract and 13 suffered from pneumonia. The 7-d and 14-d Kaplan-Meier rates of colonization were 2.21 and 7.03%. Twenty-one patterns of bronchopulmonary tract isolates were isolated from single patients and 10 from several patients. The lower airway was often the first site of colonization. The contribution of environment to patient colonization appeared to be small. The length of hospitalization, the previous use of third-generation cephalosporins less effective against P. aeruginosa, and chronic obstructive pulmonary disease were the most significant predictors of colonization/infection. The in vitro exoprotein production was not correlated with the presence of pneumonia. Our study may be useful in identifying which patients in the mechanically ventilated population are at greater risk of P. aeruginosa pneumonia.

Bacterial Proteins↗

Characterization of a macaque anti-Rh17-like monoclonal antibody.

In order to produce macaque monoclonal antibodies (mAbs) against human red blood cell (RBC) antigens, macaques were immunized with human and gorilla RBCs and their spleen lymphocytes were fused with man-mouse heteromyeloma cells. One macaque-mouse heterohybridoma produced a macaque IgGx (Cvn2-4D5) which agglutinated all human RBCs but not rare human variants Dc-,D-, and Rhnull. Thus, Cyn2-4D5 exhibited RH17-like reactivity. The specificity of Cyn2-4D5 for RHCE-encoded polypeptides was confirmed by specific immunoprecipitation of RhcE and RhCe polypeptides from human RBCs and the absence of immunoprecipitation of the RhD polypeptides extracted from D-RBCs. This study demonstrates that it is possible to produce macaque mAbs against human RBC blood group antigens.

Animals↗

Regional alteration of the transient outward current in human left ventricular septum during compensated hypertrophy.

BACKGROUND: A large calcium-insensitive transient outward current (I(to)) has been recorded in atria, left ventricular (LV) free wall, and right ventricular septal subendocardium of the human heart. Recent studies suggested a major contribution of this current to the electrical heterogeneity of the heart. However, no data have been reported on the distribution of I(to) density within the LV septal wall from compensated human LV hypertrophy. METHODS AND RESULTS: Microelectrode and patch-clamp techniques were used to record action potentials and I(to) in myocytes isolated from superficial (<3 mm deep) and deep (3 to 6 mm deep) layers of LV septum from patients with aortic stenosis and compensated LV hypertrophy. Subendocardial specimens were also obtained from undiseased donor hearts. In none of the superficial subendocardial cells from diseased hearts was a macroscopic I(to) recorded (n=42), whereas in cells from the same location from donor hearts, a typical I(to) was clearly present, with a peak density of 5.88+/-0.78 pA/pF at +60 mV (n=4). However, in deep layers from patients with compensated LV hypertrophy, macroscopic I(to) was present, with a peak density of 10.50+/-2.58 pA/pF at +60 mV (n=4). The absence of I(to) in superficial septal cells from hypertrophied hearts was not due to a divalent cation-related shift of the current kinetics. Instead, extracellular Ca2+ removal induced an I(to)-like current, possibly carried by K+ ions, with a peak density of 30.7+/-2.6 pA/pF at +60 mV (n=29). However, its magnitude, kinetics, and pharmacological characteristics did not allow identification of this current as the usual I(to). CONCLUSIONS: Both topography and pathology can be major modulating factors of the regional distribution of I(to) density in human LV septum. Therefore, they may play a prominent role in determining electrical gradients within this region from which the early depolarization vectors start and the left-to-right activation sequence of the interventricular septum proceeds.

Action Potentials↗

Steady-state captopril renography: continuous monitoring of the captopril-induced increase in 99mTc-MAG3 mean parenchymal transit time in renovascular hypertension.

Steady-state captopril renography (SSCR) is an original technique for assessing the captopril-induced increase in technetium-99m mercaptoacetyltriglycine (99mTc-MAG3) mean parenchymal transit time (MPTT) in kidneys affected with functional renal artery stenosis (RAS). The steady-state parenchymal activity achieved by constant infusion of 99mTc-MAG3 is directly linked to the MPTT of the radiopharmaceutical. This steady-state parenchymal activity was continuously monitored from 15 min before to 60 min after a single dose of captopril in order to detect possible disruption of the steady state. SSCR was performed in 11 hypertensive patients with unilateral RAS and in two with RAS of a solitary kidney before renal revascularization (RR). In four of these patients, an additional SSCR was performed after RR. Of the ten patients whose hypertension was cured or improved by RR, one presented an uninterpretable SSCR and six presented a positive SSCR on the affected side. Control SSCR performed in four of these six patients was bilaterally negative. SSCR was also bilaterally negative in the three patients who showed no blood pressure response to RR. These preliminary results tend to indicate that, in spite of the stability of pre- and post-captopril hydration and data acquisition conditions allowed by this steady-state technique, the sensitivity is lower than expected. However, the reason for the false-negative results does not seem to be inherent to SSCR.

Adult↗

Validity of a model of cultured myocardial cells for assessment of cardioplegia.

Myocardial protection is usually studied in vitro on perfused heart preparations, but never directly on cultured cardiomyocytes. We evaluated a model of cultured newborn rat cardiomyocytes to study both the cytotoxicity and the protective effect against chemical hypoxia of three cardioplegic solutions (St Thomas' I, Bretschneider, St Thomas' II) under normothermic (37 degrees C) and hypothermic (4 degrees C) conditions. Cytotoxicity was evaluated in 50% and 100% concentrations of the cardioplegic solutions with incubation times from 90 to 360 min. Myocardial protection was studied in 50% cardioplegic solution with metabolic inhibitors. Immediate and late viabilities, after 24 h of recovery in the medium, were evaluated by simultaneous staining with fluorescein diacetate and propidium iodide. At 37 degrees C, the 50% concentration of the three cardioplegic solutions did not modify cell viability. At 37 degrees C, with 360 min of incubation, the 100% concentration of the St Thomas' I and Bretschneider solutions diminished immediate viability (mean +/- SD; medium 87% +/- 2%; St Thomas' I 58% +/- 5%; Bretschneider 37% +/- 8%; St Thomas' II 89% +/- 3%) as well as late viability (medium 69% +/- 2%; St Thomas' I 32% +/- 3%; Bretschneider 24% +/- 7%; St Thomas' II 65% +/- 4%). At 4 degrees C, immediate and late viabilities were unaffected by cardioplegic solutions. At 37 degrees C, after 360 min incubation time, metabolic inhibitors diminished immediate viability to 29% +/- 1% and late viability to zero. None of the three cardioplegic solutions used at 50% concentration prevented this effect. At 4 degrees C, immediate viability was not significantly affected by metabolic inhibitors (73% +/- 10%), but the use of Bretschneider cardioplegic solution seemed to be detrimental (53% +/- 9%). On the other hand, recovery phase after pretreatment with metabolic inhibitors with or without cardioplegic solutions for 360 min significantly diminished late viability (medium 63% +/- 7%; metabolic inhibitors 17% +/- 8%; St Thomas' I 17% +/- 6%; Bretschneider 8% +/- 6%; St Thomas' II 15% +/- 3%) and again cardioplegia was inefficient. In conclusion, in this in vitro model for the study of cardioplegic solutions, only pure concentrations of the St Thomas' I and Bretschneider solutions under normothermic conditions were cytotoxic. The well-known protective effects of hypothermia against ischemia and reperfusion injury were both reproduced. Therefore, and even though cardioplegia failed to have any protective effect, probably owing to a severe metabolic inhibition, this model may be useful for studying myocardial protection.

Animals↗

Endothelial cells of the kidney vasa recta express the urea transporter HUT11.

The cell specific expression of the human urea transporter HUT11 in human and rat kidneys was investigated by immunochemistry and in situ hybridization. Using specific rabbit polyclonal antibodies directed against the N-terminal part and the C-terminal part of HUT11, we found that endothelial cells of medullary vasa recta (outer and inner medulla) express HUT11. In addition, an HUT11-related protein expressed by smooth muscle cells of cortical arterioles can be detected by the anti-HUT11 N-terminal peptide antibody but not the anti-HUT11 C-terminal peptide antibody. The endothelial expression of HUT11 was confirmed by double labeling with anti-CD31 and anti-von Willebrand factor antibodies. No HUT11-positive cells expressed alpha smooth muscle actin, Tamm Horsfall protein, cytokeratin 18, or CAM-L1, a marker of the principal cells of collecting ducts. Medullary vasa recta of developing and mature rat kidneys were also stained with the anti-HUT11 C-terminal peptide antibody. By in situ hybridization using a specific HUT11 35S-cDNA probe on human kidney sections, medullary vasa recta but not other renal structures were found to express HUT11 mRNA. We conclude that endothelial cells of medullary vasa recta express HUT11, a specific urea transporter, which may play a role in urea recycling within the kidney and in the mechanisms of urinary concentration and urea excretion.

Animals↗

Association of private isolation rooms with ventilator-associated Acinetobacter baumanii pneumonia in a surgical intensive-care unit.

OBJECTIVE: To determine the rates and routes of Acinetobacter baumanii colonization and pneumonia among ventilated patients in a surgical intensive-care unit (SICU) before and after architectural modifications. DESIGN: A nonsequential study comparing two groups of patients. All isolates from systematic and clinical samples were genotyped by pulsed-field gel electrophoresis (PFGE). Records of patients hospitalized during the first and second periods were reviewed and findings were compared. Between the two periods, the SICU was remodeled from enclosed isolation rooms and open rooms to only enclosed isolation rooms with handwashing facilities in each room. SETTING AND PATIENTS: All patients hospitalized and mechanically ventilated for more than 48 hours in the 15-bed SICU of the University Hospital of Besançon (France). RESULTS: For the first and second periods, the rates of colonization were, respectively, 28.1% and 5.0% of patients (P < 10(-7); relative risk [RR], 2.23; 95% confidence interval [CI95], 1.8-2.75) and the specific rates of bronchopulmonary (BP) colonization were, respectively, 9.1 and 0.5 per 1,000 days of mechanical ventilation (P < 10(-5). Seven major PFGE isolate types were identified, 4 of which were isolated from 44 of the 47 colonized or infected patients. Logistic regression analysis showed that colonization was not associated with patient characteristics. CONCLUSION: Conversion from open rooms to isolation rooms may help control nosocomial BP tract acquisition of A baumanii in mechanically ventilated patients hospitalized in an SICU.

Acinetobacter↗

The molecular basis of the Kidd blood group polymorphism and its lack of association with type 1 diabetes susceptibility.

The Kidd blood group locus encodes a urea transporter which is expressed on human red cells and in the kidney. This gene is located on chromosome 18q12, and evidence for linkage and association with type 1 diabetes mellitus has been reported. To investigate this further, the genetic basis for the blood group Jk(a)/Jk(b) polymorphism was first determined by sequencing reverse-transcribed reticulocyte RNAs from Jk(a+b-) and Jk(a-b+) donors. The Jk(a)/Jk(b) polymorphism was caused by a transition (G838A), resulting in a Asp280Asn amino acid substitution and an MnlI restriction fragment length polymorphism (RFLP). Using the MnlI RFLP, we found that the Jk(a)/Jk(b) polymorphism was not in linkage disequilibrium with type 1 diabetes in 228 multiplex UK and US families tested.

Adolescent↗

Successful treatment of tetralogy of Fallot with pulmonary atresia using a pulmonary homograft.

A 13-year-old boy with tetralogy of Fallot and pulmonary atresia after a Waterston and Blalock-Taussig shunt underwent successful repair using a cryopreserved pulmonary homograft as a valved extracardiac conduit. Aortic transection gave a good operative field for pulmonary angioplasty and closure of the Waterston anastomosis. The cryopreserved pulmonary homograft provided excellent handling characteristics. The postoperative course was good, and postoperative echocardiography showed good function of the valved conduit.

Adolescent↗

Doppler echocardiographic study of three different mechanical valves in the aortic position: the use of external nominal diameter indexed to body surface area.

BACKGROUND AND AIMS OF THE STUDY: Previous studies have demonstrated the benefit of indexing body surface area (BSA) with ventriculo-arterial orifice diameters (or calculated effective surface dynamics) and with nominal external diameters of valves in order to study hemodynamic profiles in vivo or determine the clinical influence of this parameter. This study analyzes the relationship between BSA and nominal external diameter of mechanical valves implanted in the aortic position. It also evaluates the potential interest of using these diameters indexed to BSA in an echo-Doppler study of valves. METHODS: During 1994, a prospective echo-Doppler study of three models of mechanical valves in the aortic position was carried out. RESULTS: The echo-Doppler parameters of the aortic St. Jude Medical, Sorin Bicarbon and Dideco Monostrut prostheses have been studied with regard to nominal external diameters and were found identical to published data. There were no significant differences in these parameters between the three valves. In 128 patients, prosthetic external diameters were evenly distributed heterogeneously with regard to BSA. Indexing of nominal external diameter with BSA appeared a more reliable reference to study different echo-Doppler parameters, minimizing the effect of cardiac output. CONCLUSION: The in vivo echo-Doppler determination at rest of valve hemodynamic profiles should be carried out with reference to the nominal external diameter indexed to the BSA.

Adult↗