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Biomedical subjects

A Joly

Publications and source records attributed to A Joly.

At least 19 recordsLinked to original sources

Assessing the within-herd prevalence of cows antibody-positive to bovine viral diarrhoea virus with a blocking ELISA on bulk tank milk.

Milk samples from 135 herds in Brittany were tested by a blocking ELISA for antibodies to bovine viral diarrhoea virus (BVDV) and used to assess the relationship between the bulk milk result and the within-herd prevalence of antibody-positive lactating cows. This relationship was first quantified by using a general linear model and controlling for the number of cows contributing milk to the bulk tank, for the percentage of primiparous cows in the herds and for the number of milkings contributing to the bulk tank. Receiver operating characteristic (ROC) analysis was then used to define classes of percentage inhibition in the bulk milk associated with minimum intraclass and maximum between-class variances of the within-herd prevalence. Only the percentage inhibition of bulk milk had a significant positive effect on within-herd prevalence (R2 = 0.85). The ROC analysis provided three classes of bulk milk results corresponding to different expected levels of within-herd prevalence. Herds with bulk milk percentage inhibitions of 0 to 35 per cent, 35 to 60 per cent and 60 to 100 per cent had mean (sd) observed prevalences of 4.8 (5.7) per cent, 21.6 (14.6) per cent and 66.0 (29.3) per cent, respectively. Herds with a bulk milk inhibition of 0 to 35 per cent were expected to be BVDV-free. A herd with two consecutive bulk milk results four months apart of 60 per cent or more was likely to have a very high prevalence (median of 93 per cent) and could be suspected of harbouring an active infection.

Animals↗

Evaluation of a blocking ELISA for the detection of bovine viral diarrhoea virus (BVDV) antibodies in serum and milk.

The performance characteristics of a blocking ELISA test applied to serum and individual milk for the detection of antibodies to bovine viral diarrhoea virus (BVDV) were assessed using 1189 matched milk/serum samples collected from cows of 42 dairy herds located in Brittany (west of France). This test was based on a monoclonal antibody directed against non-structural protein NS2-3 of pestiviruses. All tests were performed blind. For each type of sample, negative/positive cut-off values were determined using receiver operating characteristic (ROC) analysis. Sensitivity and specificity were estimated using the virus neutralisation test as a reference. For sera, the ROC analysis provided a negative/positive inhibition percentage cut-off value of 50% giving a sensitivity and a specificity of 96.9 and 97.8%. For individual milk samples, the cut-off was fixed at 30%, leading to a sensitivity and a specificity of 96.9 and 97.3%. Using this test, a good overall agreement was found between results obtained on matched milk/serum samples (Kappa value=0.95). The present results indicate that this blocking ELISA test is reliable enough for use in a mass screening and control scheme on BVDV.

Animals↗

Informative value of an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of bovine viral diarrhoea virus (BVDV) antibodies in milk.

Bulk and individual milk samples from 117 herds located in Brittany (west France) were used to assess: (i) the performance characteristics of an indirect enzyme-linked immunosorbent assay (ELISA) applied to individual milk for the detection of antibodies to bovine viral diarrhoea virus (BVDV); and (ii) the relationship between the bulk milk result obtained from this test and the within-herd prevalence of antibody-positive lactating cows. This ELISA test was based on a monoclonal antibody directed against non-structural protein NS2-3 of pestiviruses. At the individual level, based on 1113 matched milk/serum samples, the sensitivity and specificity of this test applied to milk, compared with the virus neutralization test on serum, were 95.0 and 97.7%, respectively. At the herd level, the relationship between the optical density percentage (OD%) of bulk milk and the within-herd prevalence of antibody-positive lactating cows was assessed using the receiver operating characteristics (ROC) analysis. Classes of OD% of bulk milk were determined so that they were associated with minimum intraclass and maximum between-class variances of within-herd prevalence of antibody-positive cows. The ROC analysis resulted in two classes of bulk milk results corresponding to different expected levels of within-herd prevalence. Herds with an OD% of bulk milk < 75% and > or = 75% had a mean observed prevalence of antibody-positive cows of 8.9 and 60.6%, respectively. Herds with a bulk milk result < 75% were expected to be BVDV free, whereas large variations in prevalence of antibody-positive cows existed in the herds with OD% > or = 75%. The test described in this study is suitable to identify herds likely to have a low prevalence of BVDV antibody-positive cows.

Animals↗

Pharmacokinetics and analgesic effect of ropivacaine following ilioinguinal/iliohypogastric nerve block in children.

BACKGROUND: The aim was to investigate the efficacy, tolerance and pharmacokinetics of ropivavcaine when administered for ilioinguinal/iliohypogastric block in children. METHODS: We examined the pharmacokinetics and analgesic efficacy after ilioinguinal/iliohypogastric nerve block with 3 mg.kg-1 ropivacaine 5 mg.ml-1 in 22 children, aged 1-12 years, who were scheduled for inguinal surgery. Sixteen of 22 patients had a postoperative pain score < 4 (Objective Pain Scale). Nine children were given supplementary analgesics during the first six postoperative hours. RESULTS: The peak plasma concentration of total ropivacaine was 1.50 +/- 0.93 mg.l-1 (mean +/- SD) (range 0.64-4.77 mg.l-1) 15-64 min after the injection. The peak plasma concentration of free ropivacaine was 0.05 +/- 0.03 mg.l-1 (0.02-0.14 mg.l-1), which is well below the threshold for toxicity in adults. The terminal half-life was 2.0 +/- 0.7 h. No safety concerns or symptoms suggestive of systemic toxicity were observed. CONCLUSION: A dose of 3 mg.kg-1 of ropivacaine given as a single ilioinguinal/iliohypogastric nerve block in 1-12-year-old children provides satisfactory postoperative pain relief, and is well tolerated.

Amides↗

Comparison of recovery after intermediate duration of anaesthesia with sevoflurane and isoflurane.

BACKGROUND: The purpose of this study was to compare recovery from anaesthesia after sevoflurane and isoflurane were administered to children for more than 90 min. METHODS: After parental informed consent and ethical committee approval, children aged between 2 months and 6 years, ASA I or II, were randomly allocated to sevoflurane (n=20) or isoflurane (n=20) groups. Halogenated agents were discontinued following skin closure and patients were ventilated mechanically with 100% oxygen until minimum alveolar concentration (MAC) values awake were obtained (endtidal concentrations 0.6 MAC for sevoflurane and 0.4 MAC for isoflurane). Effective perioperative analgesia was provided by a caudal block. RESULTS: The mean (+/- SD) duration of anaesthesia was 132 +/- 38 min and 139 +/- 49 min for sevoflurane and isoflurane, respectively. Early recovery occurred sooner in the isoflurane group (time to extubation was 16 +/- 7 min and 11 +/- 5 min, P<0.01; Aldrete's score at 0 min was 5.5 +/- 1.5 and 7.4 +/- 1.8, P<0.001, respectively). But the time to be fit for discharge from recovery room was similar at 136 +/- 18 min and 140 +/- 20 min, respectively. CONCLUSIONS: After intermediate duration of anaesthesia administered to children for up to 90 min, isoflurane and sevoflurane allow recovery after approximatively the same lapse of time.

Anesthesia Recovery Period↗

Vascular endothelial growth factor (VEGF121) protects rats from renal infarction in thrombotic microangiopathy.

BACKGROUND: Renal thrombotic microangiopathy, typified by the hemolytic uremic syndrome, is associated with endothelial cell injury in which the presence of cortical necrosis, extensive glomerular involvement, and arterial occlusive lesions correlates with a poor clinical outcome. We hypothesized that the endothelial survival factor vascular endothelial growth factor (VEGF) may provide protection. METHOD: Severe, necrotizing, thrombotic microangiopathy was induced in rats by the renal artery perfusion of antiglomerular endothelial antibody, followed by the administration of VEGF or vehicle, and renal injury was evaluated. RESULTS: Control rats developed severe glomerular and tubulointerstitial injury with extensive renal necrosis. The administration of VEGF significantly reduced the necrosis, preserved the glomerular endothelium and arterioles, and reduced the number of apoptotic cells in glomeruli (at 4 hours) and in the tubulointerstitium (at 4 days). The prosurvival effect of VEGF for endothelium may relate in part to the ability of VEGF to protect endothelial cells from factor-induced apoptosis, as demonstrated for tumor necrosis factor-alpha (TNF-alpha), which was shown to be up-regulated through the course of this model of renal microangiopathy. Endothelial nitric oxide synthase expression was preserved in VEGF-treated rats compared with its marked decrease in the surviving glomeruli and interstitium of the antibody-treated rats that did not receive VEGF. CONCLUSIONS: VEGF protects against renal necrosis in this model of thrombotic microangiopathy. This protection may be mediated by maintaining endothelial nitric oxide production and/or preventing endothelial cell death.

Animals↗

The effect of preoperative epidural morphine on postoperative analgesia in children.

IMPLICATIONS: We examined the effects of preoperative epidural morphine associated with general anesthesia on postoperative morphine requirements. Twenty-one children older than 6 yr scheduled for major surgery were randomly assigned to two groups, a control group and an epidural group that received a single epidural morphine injection.

Abdomen↗

Axillary block in children: single or multiple injection?

The goal of this double-blind prospective study was to compare the effect of a single injection versus multiple fractionated doses on the onset time and quality of motor and sensory block, obtained in 70 children anaesthetized with axillary block alone. The brachial plexus was identified with a peripheral nerve stimulator, and blocked with 0.5 ml.kg-1 of 1.5% lignocaine with adrenaline. In Group S (single injection), the total volume was injected after location of one nerve. In Group M (multiple fractionated doses), two nerves were located, including necessarily one nerve implicated in the surgical territory. Motor and sensory blocks were assessed according to Lanz's scale before surgery by a blinded observer. A block was considered complete if there was no feeling in at least three nerve territories at 30 min. No difference was found between groups for motor and sensory block quality. However the onset time of the block was faster after multiple fractionated doses (Group M, 25+/-7 min vs Group S, 29+/-4 min) and was faster in younger children (5-9 years: M=23+/-7 min vs S=28+/-5 min, 10-15 years: no difference). There was a significant difference in the quality of the sensory blockade of the musculocutaneous nerve: 18 versus 8 complete blocks, 10 versus 14 incomplete blocks, respectively for Group M versus Group S. No adverse effect was observed and analgesia was prolonged for more than 4 h. We can conclude that, unlike adults, fractionated doses in chilren bring no benefit to the quality of sensory and motor block. Selective block of the musculocutaneous nerve is recommended when a surgical procedure takes place in this territory.

Anesthetics, Local↗

Neutral lipid from proteinuric rat urine is a novel inhibitor of the red blood cell calcium pump.

Proteinuria may be associated with hypertension and progression of renal insufficiency, which in turn may accompany abnormalities in cell calcium homeostasis. Therefore, urine from rats made proteinuric by puromycin aminoglycoside administration was analyzed, in a search for factors affecting cellular calcium transport. Proteinuric urine was fractionated by thin-layer chromatography and HPLC, and the effects of the fractions on the plasma membrane calcium pump in human red blood cells were assessed. Proteinuric urine contained a powerful specific inhibitor of the calcium pump that had little or no effect on the Na+/K+- or Mg2+-ATPases. The inhibitor was characterized as a neutral lipid, migrating as a single band, that inhibited 45Ca2+ efflux. To confirm the presence of an inhibitor in other proteinuric states, the urine from two patients with proteinuria was examined and subjected to chromatography as in the rat studies. These thin-layer chromatographic fractions contained a very strong inhibitor of the red blood cell calcium pump, suggesting that this substance may have relevance for the pathogenesis of proteinuric renal disease in human patients. Rat proximal tubule cells in tissue culture, when challenged with lipid-replete albumin, secreted an inhibitor of the calcium pump that migrated in the same chromatographic band as the urine factor. Therefore, the processing of fatty acids borne by albumin into endocytosing proximal tubular epithelium results in the synthesis and release of a previously unknown lipid modulator of the calcium pump, an effect that may predispose kidney tissue toward elevations in cytosolic calcium levels in target cells.

Adenosine Triphosphatases↗

A new structural class of proteasome inhibitors that prevent NF-kappa B activation.

The multicatalytic proteinase or proteasome is a highly conserved cellular structure that is responsible for the ATP-dependent proteolysis of many proteins involved in important regulatory cellular processes. We have identified a novel class of inhibitors of the chymotrypsin-like proteolytic activity of the 20S proteasome that exhibit IC50 values ranging from 0.1 to 0.5 microgram/mL (0.1 to 1 microM). In cell proliferation assays, these compounds inhibit growth with an IC50 ranging from 5 to 10 micrograms/mL (10-20 microM). A representative member of this class of inhibitors was tested in other biological assays. CVT-634 (5-methoxy-1-indanone-3-acetyl-leu-D-leu-1-indanylamide) prevented lipopolysaccharide (LPS), tumor necrosis factor (TNF)-, and phorbol ester-induced activation of nuclear factor kappa B (NF-kappa B) in vitro by preventing signal-induced degradation of I kappa B-alpha. In these studies, the I kappa B-alpha that accumulated was hyperphosphorylated, indicating that CVT-634 did not inhibit I kappa B-alpha kinase, the enzyme responsible for signal-induced phosphorylation of I kappa B-alpha. In vivo studies indicated that CVT-634 prevented LPS-induced TNF synthesis in a murine macrophage cell line. In addition, in mice pretreated with CVT-634 at 25 and 50 mg/kg and subsequently treated with LPS, serum TNF levels were significantly lower (225 +/- 59 and 83 +/- 41 pg/mL, respectively) than in those mice that were treated only with LPS (865 +/- 282 pg/mL). These studies suggest that specific inhibition of the chymotrypsin-like activity of the proteasome is sufficient to prevent signal-induced NF-kappa B activation and that the proteasome is a novel target for the identification of agents that may be useful in the treatment of diseases whose etiology is dependent upon the activation of NF-kappa B.

Adenosine Triphosphatases↗

[Sever acute pulmonary edema after peri-anesthetic laryngospasm in a newborn infant].

Severe acute pulmonary oedema following peranaesthetic laryngospasm in a newborn. The authors report a case of severe acute pulmonary oedema secondary to a laryngeal spasm in a 3-week-old neonate, immediately after induction of anaesthesia with halothane. After emergency tracheal intubation, the infant experienced a severe, life-threatening pulmonary oedema requiring prolonged intensive care. Such a secondary time course is unusual. Usually pulmonary oedema has a favourable outcome after oxygen administration and maintenance of positive expiration pressure, except in the neonate.

Acute Disease↗

[Postoperative pain. Particularities in the child of less than 5 years, neonatology excluded].

For many years, postoperative pain has been undertreated in children less than 5 years old in comparison to adults. The assessment of pain is indeed difficult in this range of age, and only the scales of hetero-evaluation are used. The guidelines for treatment are similar as in adults: systematic administration, balanced analgesia, evaluation of pain and potential adverse effects. Non opioid analgesics used are mainly paracetamol, niflumic acid and ibuprofen. Morphine remains the drug of choice among opioids; however the risk of respiratory depression in higher in infants less than 3 months old. Nalbuphine is also widely used in paediatrics. In addition, regional anaesthesia, either in single shot for minor surgery, or in continuous administration through epidural catheter for major surgery, has changed the management of postoperative pain in paediatrics.

Analgesia↗

[Comparative hemodynamic effects of sevoflurane and halothane at tele-expiratory concentration in intubation of infants].

OBJECTIVE: To compare the cardiovascular changes at the end-tidal concentrations of sevoflurane versus halothane required for tracheal intubation in infants (intubation MAC). STUDY DESIGN: Prospective randomized study. PATIENTS: Thirty-two infants, ASA physical status 1 or 2, scheduled for elective surgery, randomized to receive either halothane or sevoflurane for anaesthetic induction by inhalation. METHODS: Cardiovascular and echocardiographic data were recorded in both groups at baseline, and at the end-tidal concentrations needed for intubation. RESULTS: Intubation MAC was significantly less with sevoflurane than with halothane in infants. Sevoflurane did not change heart rate (HR) and cardiac index (CI) compared to values when awake. Sevoflurane significantly decreased blood pressure, systemic vascular resistance (SVR) and shortening fraction (SF). Myocardial contractility assessed by stress-velocity index (SVI) and stress-shortening index (SSI) decreased significantly at the intubation MAC, but did not fall into the abnormal range. Halothane caused a greater decrease in HR, SF, SSI, and CI than sevoflurane. CONCLUSIONS: Sevoflurane decreases cardiac output less than halothane in infants at the intubation MAC, due to a lower end-tidal concentration at intubation MAC and to less effects on haemodynamic variables.

Anesthetics, Inhalation↗

CVT-313, a specific and potent inhibitor of CDK2 that prevents neointimal proliferation.

The activity of cyclin-dependent kinase 2 (CDK2) is essential for progression of cells from G1 to the S phase of the mammalian cell cycle. CVT-313 is a potent CDK2 inhibitor, which was identified from a purine analog library with an IC50 of 0.5 microM in vitro. Inhibition was competitive with respect to ATP (Ki = 95 nM), and selective CVT-313 had no effect on other, nonrelated ATP-dependent serine/threonine kinases. When added to CDK1 or CDK4, a 8.5- and 430-fold higher concentration of CVT-313 was required for half-maximal inhibition of the enzyme activity. In cells exposed to CVT-313, hyperphosphorylation of the retinoblastoma gene product was inhibited, and progression through the cell cycle was arrested at the G1/S boundary. The growth of mouse, rat, and human cells in culture was also inhibited by CVT-313 with the IC50 for growth arrest ranging from 1.25 to 20 microM. To evaluate the effects of CVT-313 in vivo, we tested this agent in a rat carotid artery model of restenosis. A brief intraluminal exposure of CVT-313 to a denuded rat carotid artery resulted in more than 80% inhibition of neointima formation. These observations suggest that CVT-313 is a promising candidate for evaluation in other disease models related to aberrant cell proliferation.

Animals↗

Cyclosporine A is an uncompetitive inhibitor of proteasome activity and prevents NF-kappaB activation.

Cyclosporine A is an immunosuppressive agent that is used clinically in the prevention of transplant rejection and development of graft-versus-host disease. Recently, cyclosporine A has been shown to possess anti-inflammatory properties and is capable of inhibiting lipopolysaccharide-induced NF-kappaB activation. Ubiquitin-mediated proteasomal proteolysis plays a critical role in signal-induced NF-kappaB activation since it regulates both IkappaB degradation and p105 processing, it is also involved in the production of peptides for the assembly of MHC class I molecules. We report here that cylcosporine A acts as an uncompetitive inhibitor of the chymotrypsin-like activity of the 20S proteasome in vitro and that it suppresses lipopolysaccharide-induced IkappaB degradation and p105 processing in vivo demonstrating that inhibition of proteasome proteolysis is the mechanism by which cyclosporine A prevents NF-kappaB activation. A structurally unrelated immunosuppressant, rapamycin, did not inhibit the 20S proteasome in vitro.

Animals↗

The involvement of an LPS inducible I kappa B kinase in endotoxin tolerance.

When human ovarian carcinoma cells are challenged with endotoxin, an I kappa B kinase is transiently induced within 3 to 5 min. This enzyme activity causes the hyperphosphorylation and subsequent degradation of I kappa B which allows NF-kappa B to translocate to the nucleus where it activates transcription. When endotoxin treated cells are rechallenged with a second dose of LPS, I kappa B kinase is not detected and I kappa B remains in the cytoplasm where it sequesters NF-kappa B. We report here the absence of endotoxin inducible I kappa B kinase activity in endotoxin tolerant cells suggesting that I kappa B kinase may play an important role in endotoxin tolerance. When cells tolerant to endotoxin are treated with TNF, I kappa B kinase activity is induced. Thus cells that are endotoxin tolerant are not cross tolerant to TNF. Dexamethasone, a known inhibitor of NF-kappa B activation does not inhibit endotoxin dependent induction of I kappa B kinase suggesting that the mechanism of action of dexamethasone is different from the tolerance mechanism reported here.

Base Sequence↗

Survival, proliferation, and functions of porcine hepatocytes encapsulated in coated alginate beads: a step toward a reliable bioartificial liver.

Orthotopic liver transplantation is the most effective treatment for fulminant hepatic failure. As an alternative treatment, an efficient extracorporeal bioartificial liver should contain a large yield of functional hepatocytes with an immunoprotective barrier, for providing temporary adequate metabolic support to allow spontaneous liver regeneration or for acting as a bridge toward transplantation. Survival, proliferation, and functions of porcine hepatocytes were evaluated in primary cultures and after embedding in alginate beads, which were subsequently coated with a membrane made by a transacylation reaction between propylene glycol alginate and human serum albumin. Disruption of total pig livers by collagenase perfusion/recirculation allowed the obtention of up to 10(11) hepatocytes with a viability greater than 95%. Hepatocytes in conventional cultures or embedded in coated alginate beads survived for about 10 days, secreted proteins, particularly albumin, and maintained several phase I and II enzymatic activities, namely ethoxyresorufin-O-deethylase, oxidation of nifedipine to pyridine, phenacetin deethylation to paracetamol, glucuroconjugation of paracetamol, and N-acetylation of procainamide. Typical features of mitosis and [3H]thymidine incorporation indicated that porcine hepatocytes proliferated in both conventional cultures and alginate beads. The efficacy of the membrane surrounding alginate beads for protecting cells from immunoglobulins was tested by embedding HLA-typed human lymphocytes, which were subsequently incubated with specific anti-HLA immunoglobulin G and complement. These data show that large yields of porcine hepatocytes that are embedded in coated alginate beads remain functional and are isolated from large molecular weight molecules, such as immunoglobulins. This system represents a promising tool for the design of an extracorporeal bioartificial liver, containing xenogeneic hepatocytes, to treat acute liver disease in humans.

Acetaminophen↗