KIRREL is differentially expressed in adipose tissue from 'fertil+' and 'fertil-' cows: in vitro role in ovary?

We have previously shown that dairy cows carrying the 'fertil-' haplotype for one quantitative trait locus affecting female fertility located on the bovine chromosome three (QTL-F-Fert-BTA3) have a significantly lower conception rate and body weight after calving than cows carrying the 'fertil+' haplotype. Here, we compared by Tiling Array the expression of genes included in the QTL-F-Fert-BTA3 in 'fertil+' and 'fertil-' adipose tissue one week after calving when plasma non-esterified fatty acid concentrations were greater in 'fertil-' animals. We observed that thirty-one genes were overexpressed whereas twelve were under-expressed in 'fertil+' as compared to 'fertil-' cows (P < 0.05). By quantitative PCR and immunoblot we confirmed that adipose tissue KIRREL mRNA and protein were significantly greater expressed in 'fertil+' than in 'fertil-'. KIRREL mRNA is abundant in bovine kidney, adipose tissue, pituitary, and ovary and detectable in hypothalamus and mammary gland. Its expression (mRNA and protein) is greater in kidney of 'fertil+' than 'fertil-' cows (P < 0.05). KIRREL (mRNA and protein) is also present in the different ovarian cells with a greater expression in granulosa cells of 'fertil+' than 'fertil-' cows. In cultured granulosa cells, recombinant KIRREL halved steroid secretion in basal state (P < 0.05). It also decreased cell proliferation (P < 0.05) and in vitro oocyte maturation (P < 0.05). These results were associated to a rapid increase in MAPK1/3 and MAPK14 phosphorylation in granulosa cells and to a decrease in MAPK1/3 phosphorylation in oocyte. Thus, KIRREL could be a potential metabolic messenger linking body composition and fertility.

Lipid-based nutrient supplements do not affect efavirenz but lower plasma nevirapine concentrations in Ethiopian adult HIV patients.

OBJECTIVES: Lipid-based nutrient supplements (LNSs) are increasingly used in HIV programmes in resource-limited settings. However, the possible effects of LNSs on the plasma concentrations of antiretroviral drugs have not been assessed. Here, we aimed to assess the effects of LNSs on plasma efavirenz and nevirapine trough concentrations in Ethiopian adult HIV-infected patients. METHODS: The effects of LNSs were studied in adults initiating antiretroviral therapy (ART) in a randomized trial. Patients with body mass index (BMI) > 17 kg/m(2) (n = 282) received daily supplementation of an LNS containing whey (LNS/w), an LNS containing soy (LNS/s) or no LNS. Trough plasma concentrations of efavirenz and nevirapine were measured at 1 and 2 months. Genotyping for 516 G>T and 983 T>C polymorphisms of the cytochrome P450 (CYP) 2B6 locus was performed. Multilevel linear mixed-effects models were used to assess the associations between LNS and plasma efavirenz and nevirapine concentrations. RESULTS: In patients with BMI > 17 kg/m(2), nevirapine concentrations were lower in the LNS/w and LNS/s groups by a median of -2.3 µg/mL [interquartile range (IQR) -3.9; -0.9 µg/mL; P = 0.002] and -2.1 µg/mL (IQR -3.9; -0.9 µg/mL; P = 0.01), respectively, compared with the group not receiving supplements. There were no differences between groups with respect to efavirenz plasma concentrations. The CYP2B6 516 G>T polymorphism was associated with a 5 µg/mL higher plasma efavirenz concentration compared with the wild type (P < 0.0001), while it was not associated with plasma nevirapine concentrations. CONCLUSIONS: Intake of an LNS was associated with lower plasma nevirapine trough concentrations, indicating possible drug-LNS interactions. The clinical relevance of such reductions in nevirapine exposure is not clear. Plasma efavirenz concentration was not affected by the LNS.

Pharmacokinetics of ofloxacin and levofloxacin for prevention and treatment of multidrug-resistant tuberculosis in children.

Limited data on fluoroquinolone pharmacokinetics and cardiac effects in children exist. Among 22 children receiving drug-resistant tuberculosis prophylaxis or treatment, serum concentrations following oral doses of levofloxacin (15 mg/kg of body weight) and ofloxacin (20 mg/kg) were lower than those expected from existing pediatric data, possibly due to differences in the formulations (crushed tablets). Drug exposures were lower than those in adults following standard doses and below the proposed pharmacodynamic targets, likely due to more rapid elimination in children. No QT prolongation was observed.

[Role of the fatty acids in ovarian functions: involvement of peroxisome proliferator-activated receptors (PPAR) and adipokines]. / Rôle des acides gras sur les fonctions ovariennes : implications des Peroxisome Proliferator-Activated Receptors (PPAR) et des adipocytokines.

The impact of nutrition and energy reserves on the reproductive functions is known for a very long time. However, the metabolic factors involved in the interactions between nutrition and reproduction are still poorly understood. These factors may be hormones or nutrients (glucose, protein and fatty acids). However, it remains to determine whether these factors act directly or indirectly on the reproductive tissues. In this issue, we briefly summarize the impact of fatty acids on the development of ovarian follicles, oocyte and embryo. We then discuss the current hypotheses about the mechanisms of action of these fatty acids on the ovarian functions. We describe more particularly the role of some receptors of fatty acids, Peroxisome Proliferator-Activated Receptors (PPAR) and Liver X Receptors (LXR) and two adipokines, leptin and adiponectin on ovarian cells.

Ligand binding to heparan sulfate proteoglycans induces their aggregation and distribution along actin cytoskeleton.

Cell surface heparan sulfate proteoglycans (HSPGs) participate in molecular events that regulate cell adhesion, migration, and proliferation. The present study demonstrates that soluble heparin-binding proteins or cross-linking antibodies induce the aggregation of cell surface HSPGs and their distribution along underlying actin filaments. Immunofluorescence and confocal microscopy and immunogold and electron microscopy indicate that, in the absence of ligands, HSPGs are irregularly distributed on the fibroblast cell surface, without any apparent codistribution with the actin cytoskeleton. In the presence of ligand (lipoprotein lipase) or antibodies against heparan sulfate, HSPGs aggregate and colocalize with the actin cytoskeleton. Triton X-100 extraction and immunoelectron microscopy have demonstrated that in this condition HSPGs were clustered and associated with the actin filaments. Crosslinking experiments that use biotinylated lipoprotein lipase have revealed three major proteoglycans as binding sites at the fibroblast cell surface. These cross-linked proteoglycans appeared in the Triton X-100 insoluble fraction. Platinum/carbon replicas of the fibroblast surface incubated either with lipoprotein lipase or antiheparan sulfate showed large aggregates of HSPGs regularly distributed along cytoplasmic fibers. Quantification of the spacing between HSPGs by confocal microscopy confirmed that the nonrandom distribution of HSPG aggregates along the actin cytoskeleton was induced by ligand binding. When cells were incubated either with lipoprotein lipase or antibodies against heparan sulfate, the distance between immunofluorescence spots was uniform. In contrast, the spacing between HSPGs on fixed cells not incubated with ligand was more variable. This highly organized spatial relationship between actin and proteoglycans suggests that cortical actin filaments could organize the molecular machinery involved in signal transduction and molecular movements on the cell surface that are triggered by heparin-binding proteins.

Therapeutic drug monitoring of amlodipine and the Z-FHL/HHL ratio: Adherence tools in patients referred for apparent treatment-resistant hypertension.

BACKGROUND: Non-adherence to antihypertensives is a cause of 'pseudo-treatment-resistant' hypertension. OBJECTIVE: To determine whether monitoring plasma amlodipine concentrations and inhibition of angiotensin-converting enzyme (ACE) can be adjunct adherence tools. METHODS: Patients with hypertension who were prescribed enalapril and amlodipine were enrolled. Blood pressures (BPs) were monitored and an adherence questionnaire was completed. Steady-state amlodipine was assayed using liquid chromatography-mass spectrometry and degree of ACE inhibition using the Z-FHL/HHL (z-phenylalanine-histidine-leucine/hippuryl-histidine-leucine) ratio. RESULTS: One hundred patients (mean (standard deviation) age 50.5 (12) years, 46% male) were enrolled. Based on plasma assays, 26/97 patients (26.8%) were unsuppressed by enalapril and 20/100 (20%) were sub-therapeutic for amlodipine. There were significant BP differences based on plasma levels of the medication: 21/20 mmHg lower in the group with suppressed ACE and 26/20 mmHg in the group with steady-state amlodipine concentrations. CONCLUSIONS: Monitoring antihypertensive adherence by assaying plasma medication concentrations is a feasible option for evaluating true v. pseudo-resistant hypertension.

Cyclic Vomiting Syndrome is characterized by altered functional brain connectivity of the insular cortex: A cross-comparison with migraine and healthy adults.

Cyclic Vomiting Syndrome (CVS) has been linked to episodic migraine, yet little is known about the precise brain-based mechanisms underpinning CVS, and whether these associated conditions share similar pathophysiology. We investigated the functional integrity of salience (SLN) and sensorimotor (SMN) intrinsic connectivity networks in CVS, migraine and healthy controls using brain functional Magnetic Resonance Imaging. CVS, relative to both migraine and controls, showed increased SLN connectivity to middle/posterior insula, a key brain region for nausea and viscerosensory processing. In contrast, this same region showed diminished SMN connectivity in both CVS and migraine. These results highlight both unique and potentially shared pathophysiology between these conditions, and suggest a potential target for therapeutics in future studies.

Alpha v integrin antagonists induce the disassembly of focal contacts in melanoma cells.

In recent years, several antagonists of alpha(v)beta3 have been used to develop therapeutic approaches to the treatment of melanoma neoplasia. We studied the effects of anti-alpha(v)-integrin-blocking antibodies on attached M21 melanoma cells, the cellular distribution of alpha(v)-integrin and the molecular organization of focal structures. Anti-alpha(v)-integrin-blocking antibodies 17E6 and LM609, and an anti-alpha(v)beta3-integrin antagonist peptide cRGD 85189 induced detachment of M21 melanoma cells cultured for 24 hours on various substrates. cRGD was the most effective antagonist, reducing the number of adherent cells by 80%, while 17E6 reduced adhesion by only 30%. Light- and electron microscopy revealed attached cells with a flat shape and well-formed actin cytoskeleton. After treatment, cells became rounded and detached from the culture dish. alpha(v)-Integrins and focal-contact proteins were observed at adhesion sites in focal structures by immunocytochemistry. After treatment, however, cell rounding was accompanied by disorganization of the actin filaments and redistribution of alpha(v)-integrins and most of the focal proteins studied, except vinculin and tensin. Our results indicate that treatment of M21 melanoma cells with a(v)-integrin antagonists disrupts the actin cytoskeleton, redistributes a(v)-integrin and induces molecular disassembly of focal contacts.

Intracellular distribution of hepatic glucokinase and glucokinase regulatory protein during the fasted to refed transition in rats.

We have studied the intracellular distribution in vivo of glucokinase (GK) and glucokinase regulatory protein (GKRP) in livers of fasted and refed rats, using specific antibodies against both proteins and laser confocal fluorescence microscopy. GK was found predominantly in the nucleus of hepatocytes from starved rats. GK was translocated to the cytoplasm in livers of 1- and 2-h refed animals, but returned to the nucleus after 4 h. GKRP concentrated in the hepatocyte nuclei and its distribution did not change upon refeeding. These results show that, in physiological conditions, GKRP is present predominantly in the nuclei of hepatocytes and that the translocation of hepatic GK from and to the nucleus is operative in vivo.

Comparison of risk of orthostatic hypotension in elderly depressed hypertensive women treated with nortriptyline and thiazides versus elderly depressed normotensive women treated with nortriptyline.

This is a nonblind, case-controlled study comparing the risk of orthostatic hypotension (OH) in 2 groups of elderly depressed women: 22 normotensive and 21 hypertensive patients receiving thiazides. Blood pressure measurements and tilt-table tests produced similar results: increased drop in systolic blood pressure (SBP) after standing (p <0.001), with no significant differences between the groups (p = 0.523). There were no changes on diastolic blood pressure (DBP) after standing, or in SBP or DBP at rest. Dizziness was reported by 23 subjects (53.5%) before treatment, and by 16 subjects (38.1%) at week 8. Complaints of dizziness were not associated with OH (Kappa = 0.07).

Heparin-induced thrombocytopenia and thrombosis following open heart surgery.

We recently observed five cases of early thrombus formation in patients undergoing anticoagulation with subcutaneous heparin following open heart surgery. The reasons prompting surgery were as follows: one mitral valve replacement, one double valve replacement, one mitral valve reconstruction, one aortic valve replacement associated with coronary bypass. In all cases, intravenous heparin was begun on the day of surgery and replaced by subcutaneous (SC) heparin on postoperative day 1. Acute thrombocytopenia was observed between the 6th and 11th postoperative day. This was interpreted as denoting an idiosyncratic reaction to heparin which was replaced by low molecular weight heparin (LMWH) in two cases and by acenocoumarol in the other cases. Massive thrombosis of the aortic valve resulted in the death of one patient. Thrombosis of the left atrium occurred in three patients (two of whom had a transient ischemic attack (TIA)). One patient had aorto-iliac thrombosis. Successful reoperation was carried out in four of the five patients. Although heparin-induced thrombocytopenia and thrombosis [HITT] is a rare complication of heparin therapy, serial platelet count monitoring and in vitro platelet aggregation tests are mandatory in the diagnosis of this syndrome. Discontinuation of heparin is indicated as soon as the syndrome is recognized and the institution of aspirin is recommended if the thromboembolic complication requires reoperation and reexposure to heparin.

Internal features of graphite in cast irons. Confocal microscopy: useful tool for graphite growth imaging.

Spherulitic crystallisation is a mode of growth of crystals from the melt. Considerable attention has been given to spheroidal graphite formation, providing detailed information about the internal microstructure of the spherulites in spheroidal (SG irons) and compacted graphite irons (CG irons) (Stefanescu, D., 1990. Cast Irons. ASM Handbook, 10th ed., vol. 1). Nevertheless, the mechanisms responsible for this mode of crystallisation are not fully understood. This study deals with the inoculation mechanisms, with particular emphasis on the study of the inclusions for the heterogeneous nucleation of graphite. It is shown that the graphite nuclei are sulfide products of the nodularizing treatment. It has been observed that when rare-earth treatment is applied, the central nucleus consists of a core and an envelope from which the graphite grows. Confocal Scanning Laser Microscopy (CSLM), in reflection mode, was used to study the internal features of the spheroidal graphite growth. Confocal reflection imaging, which has a capacity for optical sectioning of the sample, provides high-resolution images of surface and subsurface regions of interest contained within a semi-transparent sample. Furthermore, three-dimensional reconstruction of these optical sections can provide insight into the mechanism of graphite growth mechanism interpretation. With CSLM the radial growth of graphite was seen. Other techniques, such as TEM, SEM-EDS, WDS, AES and SAM were also used to corroborate the results.

Cardioprotective effect of trimetazidine during coronary artery graft surgery.

Reperfusion injury remains the most uncontrolled phenomenon during cardiac surgery. Potential myocardial protection by trimetazidine was tested in a double blind placebo controlled study on 19 patients undergoing aorto-coronary bypass surgery. The trimetazidine group was composed of 10 patients and the placebo group of 9 patients. Pretreatment was started three weeks before surgery with 1 tablet (trimetazidine 20 mg) t.i.d. and the same drug was added to the cardioplegic solutions (trimetazidine: 10(-6) M). The cross clamping time was 41.1 +/- 3.8 minutes in the trimetazidine group and 39.8 +/- 2.3 minutes in the placebo group. Metabolic measurements showed that the increase of malondialdehyde measured in the coronary sinus 20 minutes after reperfusion was significantly (p = 0.014) less in the trimetazidine group (from 1.60 +/- 0.11 to 1.79 +/- 0.2 mumol/L-1) than in the placebo group (from 1.17 +/- 0.11 to 2.84 +/- 0.58 mumol/L-1). Myosin was present 4 hours after surgery in all patients in the placebo group and in 5 of the 10 of the trimetazidine group (p = 0.036). Haemodynamic measurements showed that patients pretreated with trimetazidine had a better ventricular function, as assessed by the stroke work index (SWI) significantly (p = 0.01) higher in the trimetazidine group (0.0391 +/- 0.0029 g/min/m2/beta) than in the placebo group (0.0282 +/- 0.0026 g/min/m2/beat), the evolution of SWI during surgery was not significantly different between the two groups. Thus trimetazidine seems to reduce ischaemia-reperfusion damage during cardiac surgery; moreover pretreatment with trimetazidine allows the patient to face the operation with better ventricular function.

[Heart surgery and human immunodeficiency virus]. / Chirurgie cardiaque et virus de l'immunodéficience humaine.

The authors report their experience of cardiac surgery in 9 carriers of the human immunodeficiency virus (HIV). Eight HIV seropositive patients underwent surgery under cardiopulmonary bypass for valve repair or replacement. Eight patients were asymptomatic with respect to their viral infection: one patient had generalised lymphadenopathy. The hospital mortality was 1/9 (11.1%). There was no infectious morbidity. Five patients are alive with an average follow-up of 29.6 months and in NYHA Stages I or II. One patient deteriorated and presented symptoms of an AIDS-related complex. These results suggest that if the operative indications are justified, cardiac surgery under cardiopulmonary bypass may be performed in asymptomatic HIV seropositive patients.

[Fibrinolytic inhibitors and prevention of bleeding in cardiac valve surgery. Comparison of tranexamic acid and high dose aprotinin]. / Antifibrinolytiques et prévention du saignement en chirurgie cardiaque valvulaire. Comparaison de l'acide tranexamique à l'aprotinine à haute dose.

In order to assess the effects of tranexamic acid in comparison to the high dose regimen of aprotinin recommended by Royston and considered to be the reference in postoperative bleeding in cardiac surgery, 35 consecutive patients were randomised to two groups according to the product prescribed. The global postoperative bleeding was comparable in the two groups (p = 0.49). One surgical reoperation for haemostasis was required in the reference group. There was one case of renal failure in the same group due to cardiac failure. No thrombotic complications were observed. Platelet function, as judged by the bleeding time and platelet aggregation to ristocetin, was the same in the two groups. The D-dimers remained low in both groups, reflecting the absence of intravascular coagulation and fibrinolysis. Tranexamic acid was as effective and as safe as high dose aprotinin. These two substances, in addition to their fibrinolytic inhibitory activity, conserved platelet protection by blocking the action of plasmin. These results seem to justify the preventive use of tranexamic acid from the moment of skin incision, especially in reoperation.

[Advantages and limits of programmed autologous transfusion in cardiovascular surgery. Apropos of 524 patients]. / Avantages et limites de la transfusion autologue programmée en chirurgie cardiovasculaire. A propos de 524 patients.

Between October 1987 and July 1989, 544 patients, candidates for cardiovascular surgery, were included in a trial of programmed autologous autotransfusion. Five hundred and twenty four patients underwent one or several (maximum 4) blood donation sessions in the 3 weeks before surgery with no complications. Overall, 57% of patients benefited from homologous blood transfusion, thereby avoiding all risk of contamination. It was in the group of patients able to undergo 3 or 4 preoperative blood donations that we observed the smallest number of homologous transfusions (30%). Programmed autologous transfusion would seem to be a very useful technique for cardiac surgery, allowing a reduction in health care costs without additional patient risk. In order to improve on this method, it may be useful to associate a peroperative technique of blood recuperation in patients in whom the transfusion needs are likely to exceed the possibilities of preoperative blood donation alone.

[Pre- and intraoperative management of heparin cross hypersensitivity. Apropos of a case]. / Prise en charge pré- et peropératoire d'une allergie croisée à l'héparine. A propos d'un cas.

A patient with a Starr prosthetic heart valve for 13 years developed chronic idiopathic thrombocytopaenic purpura and a highly probable crossed allergy to heparin. As the valve needed to be replaced, cardiopulmonary bypass surgery was undertaken associating heparin with lioprost and Aprotinine. In this type of situation, aggregation of control platelets by the patient's plasma in the presence of unfractionated heparin and of low molecular weight heparin justifies the use of powerful antiplatelet agents such as lioprost which was associated with Aprotinine for its platelet protective effects. This original combination allowed successful cardiopulmonary bypass surgery under unfractionated heparin under excellent conditions with minimal blood loss. This case underlines the value of this approach for cardiopulmonary bypass surgery in patients with heparin-induced thrombocytopaenia.

[Low molecular weight heparin in extracorporeal circulation. 1st clinical applications]. / Etude d'une héparine de bas poids moléculaire au cours des circulations extracorporelles. Premières applications cliniques.

The haemorrhagic complications inherent to the use of heparin during cardiac surgery led us, after a pilot experimental study, to try out a low molecular weight heparin (LMWH), PK 10169, which has weaker haemorrhagic effects in vitro. Our initial experience was confined to 23 patients with differing pathologies, undergoing cardiopulmonary bypass lasting 30 to 165 minutes. The modes of injection of YK 10169 varied according to the results, especially with respect to the limitation of peaks of anti-Xa activity; 8 patients were given one bolus intravenous injection, 9 were given a bolus injection and a continuous infusion, and 6 were only given the continuous infusion. Biological monitoring of anticoagulation was based on anti-Xa activity. Analysis of the biological results showed that the principal feature was the partial correction, and occasionally the non-correction of anti-Xa activity by protamine sulphate, with no correlation between this anti-Xa activity and postoperative bleeding. The authors report cases of severe postoperative bleeding despite the supposed theoretical and experimental weakly haemorrhagic properties of LMWH, and also discuss the inefficacy of protamine sulphate. The indications for LMWH for cardiopulmonary bypass which were retained, were the rare cases of heparin-induced thrombocytopaenia. In conclusion, it is possible to use LMWH during cardiac surgery but we do not advise using it routinely as its theoretical advantages are not confirmed in practice.

[Bacterial infections after heart surgery under extracorporeal circulation. Effects of autologous and homologous transfusions]. / Infections bactériennes après chirurgie cardiaque sous circulation extra-corporelle. Effets des transfusions autologues et homologues.

To verify that the rate of post operative infections is higher with homologous transfusion rather than without, 86 coronary artery bypass patient charts were retrospectively studied. Inclusion criteria were those of the autologous transfusion group. Four groups were defined : Gr 1 : No transfusion. Gr 2: Autotransfusion. Gr 3: Allotransfusion. Gr 4: Autotransfusion who also received homologous blood products. Although not significant, the bacterial infection rate was twofold higher in the Gr 3, than in the autologous group and was almost four times higher and significant (p < 0.01) when patients who received homologous blood products were compared to those who did not. These preliminary results lead us to enlarge our autotransfusion program and us encourage, to search for the cause of the decreased infection rate after autologous transfusion by prospective studies.

[Tolerance and efficacy of delayed autologous transfusion in cardiovascular surgery]. / Tolérance et efficacité de la transfusion autologue différée en chirurgie cardiovasculaire.

Over a period of 18 months, 313 patients (mean age 52 years) undergoing elective cardiovascular surgery were included in the autologous transfusion program involving two different Transfusion Centres. A further 10 patients were excluded because of anaemia (haemoglobin levels less than 11 g.dl-1) (n = 3), angina pectoris less than 8 days before (n = 3), patient refusal (n = 2), pneumonia (n = 1), and severe aortic insufficiency (n = 1). A maximum of 5 ml.kg-1 of blood was obtained during the 3 to 4 weeks prior to surgery, one donation being taken a week. In one Transfusion Centre, the blood was taken without tourniquet, and without any fluid replacement. Diuretics and converting enzyme inhibitors were stopped. In the opposite, in the other Centre, blood was taken using a tourniquet, and replaced by a gelatin solution (Plasmion). All the patients were given iron. The blood units were kept by the Transfusion Centres under the same conditions as homologous blood, but in a separate circuit. The 313 patients predeposited a mean of 2.71 units of blood: 4 units where obtained in 59 patients, 3 in 113, 2 in 133 and only 1 in 8. Mean haemoglobin level on starting the program was 14.49 g.dl-1. Neither homologous red cells nor plasma was administered in 176 patients (56.23%); among the 172 patients who predeposited 3 or 4 units, 123 (71.5%) were given their own blood only. Intraoperative blood salvage was used in 189 out of 313 patients (60.4%), and intraoperative haemodilution with albumin was used in 173 patients.(ABSTRACT TRUNCATED AT 250 WORDS)