Methodology for the evaluation of drugs in pregnant women

Although drugs are prescribed during pregnancy with some reluctance, they fulfill a real need in some circumstances. Adequate drug evaluation is thus essential, either based on efficacy and safety or mainly safety, using available data from non-pregnant women. Evaluation methodology is not fundamentally different during pregnancy. Recommendations for drug development are formulated on the basis of the most common situations as well as specific suggestions, thus raising the awareness of the different partners participating in healthcare (institutions, the pharmaceutical industry and prescribers). In particular, regulatory and economic incentives superimposed upon those recommendations adopted in Europe and the US for orphan diseases should be put into place to assist in the evaluation of drugs used in obstetrics. Medical needs in obstetrics should be better identified, and labelling of drugs for use during pregnancy should be better directed towards prescribers; a national registry of pregnancies should be established in France.     

Detecting life-threatening lactic acidosis related to nucleoside-analog treatment of human immunodeficiency virus-infected patients,and treatment with L-carnitine

OBJECTIVE: Our first objective was to determine a blood lactate threshold predictive of survival in human immunodeficiency virus patients experiencing lactic acidosis related to nucleoside analogs, and second, to test l-carnitine for the treatment of patients exceeding that threshold. DESIGN: a) Retrospective study using data from personal and published observations to determine the lactate threshold between survivors and nonsurvivors in human immunodeficiency virus patients being treated with nucleoside analogs. b) Prospective multicenter open trial to test l-carnitine treatment of human immunodeficiency virus patients receiving nucleoside analogs. SETTING: Medical intensive care units of four teaching hospitals and one general hospital. PATIENTS: Retrospective analysis of data from 39 human immunodeficiency virus patients (five personal cases and 34 patients from the literature) receiving nucleoside-analog treatment from which lactate values were available. An additional six patients with high lactate values were included as a pilot study testing the use of l-carnitine therapy. MEASUREMENTS AND MAIN RESULTS: An initial lactate level of 9 mmol/L, which gave good positive and negative predictive values, was determined as a threshold between survivors and nonsurvivors for the patients receiving nucleoside-analog treatment. Six patients with initial lactate levels >10 mmol/L were prospectively treated with l-carnitine; three survived beyond the end of the study. CONCLUSIONS: The blood lactate levels in human immunodeficiency virus patients receiving nucleoside-analog therapy can predict mortality in these patients. The preliminary data from this pilot study suggest that l-carnitine may be helpful for patients who have nucleoside-analog-related lactic acidosis with blood lactate levels >10 mmol/L. Further studies will be necessary to affirm the therapeutic efficacy of l-carnitine in this setting.     

Apoptosis and proliferation kinetics of T cells in patients having experienced antiretroviral treatment interruptions

Multiple failures of antiretroviral treatments, as a result of multidrug-resistant virus, have led to a proposal for structured therapeutic interruptions (STI). However, a significant decrease in CD4+ T cells may occur. The aim of our study was to determine the kinetics of T cell subpopulation changes, T cell apoptosis and peripheral blood mononuclear cell proliferation after STI. The impact of resistance mutation disappearance on T cell apoptosis was also studied. Ten patients were enrolled prospectively, and blood sampling was performed at weeks 0, 2, 4, 6, 8 and 12. The mean increase in viral load was 1.3 log(10) copies/ml, ranging from 0.1 to 3.2. CD4+ T cell count decreased to a mean of 80 cells/mm(3) from baseline to week 12. In the same period, CD8+ T cells decreased to a mean of 139 cells/mm(3). A significant increase in both T cell apoptosis and proliferation of mononuclear cells was observed. However, proliferation was an early and brief event. The increase in CD4+ T cell apoptosis was obvious in patients exhibiting complete reversion of resistance mutations to antiviral drugs. Our results suggest that during STI, apoptosis is an overwhelming phenomenon compared with proliferation, and may explain the limited immunological impact of this therapeutic option.     

Serotonin and experimental pain in healthy young volunteers

OBJECTIVE: The role of serotonin in the modulation of nociceptive input has been widely studied, and a link between serum serotonin (S-5HT) and pain thresholds elicited in patients with chronic painful pathologies has been shown. In the light of contradictory concepts on pain message modulation by S-5HT, this study tries to define whether S-5HT displays a nociceptive or antinociceptive role in experimental pain evaluation in healthy volunteers. DESIGN: In 20 healthy young volunteers, 10 men and 10 women (21 +/- 2 years old), blood serotonin measurements were made, pressure pain thresholds was determined, and statistical analysis was performed. RESULTS: This study showed a significant negative correlation of total blood serotonin with experimental pain detection threshold (P < 0.05, r = 0.444), but not with pain tolerance, while sex-related correlations of serotonin and thresholds were not significant. Lower serotonin concentration (P = 0.02), higher pain threshold (P < 0.01), and higher pain tolerance (P = 0.02) in men than in women were observed. CONCLUSION: Low pain detection thresholds may be explained by a peripheral nociceptive effect of serotonin. Pain tolerance does not, however, encompass a similar pattern of serotoninergic involvement in pain control and may include other components that remain to be elucidated. These results call for further studies on a larger population.     

Study of a chitin-based gel as injectable material in periodontal surgery.

The optimal conditions of injection of a chitin gel for applications in periodontal surgery have been studied as a function of various parameters. They correspond to a time of 2 min 15s (+/- 15"). They are achieved for acetylation parameters corresponding to: a molar ratio acetic anhydride/glucosamine residue, R = 1.5, a temperature of the master solution of 12 degrees C, a mixture of hydroalcoholic solution/acetylating reactive stirred for 45s at room temperature and a chitin concentration of 3.6%. This concentration allows us to limit the syneresis. to improve the mechanical properties of the gel and to obtain a viscosity suitable for the injection. Doping of the gel by means of chitosan powder insoluble under these conditions allows us to consider an improvement of the biological activity of the gel.     

Expression analysis of recombinant lysyl oxidase (LOX) in myofibroblastlike cells

Lysyl oxidase (LOX), originally known as the enzyme required for initiation of covalent cross-linking in collagens and elastin, is now known to be a member of a family of genetically related proteins. LOX, or a related protein, has also been localized intracellularly, both in association with the cytoskeleton and in the cell nucleus. To determine the structural requirements for secretion, maturation, and nuclear location of LOX in a cellular context, we have devised an homologous cell model for expression of the recombinant protein. Murine recombinant LOX was expressed in 3T6-5 myofibroblast-like cells as a 51-kD precursor, which was observed in the cytoplasm but not in the nucleus. To investigate whether potential alternative translation initiation sites were involved in specifying a nuclear form of LOX, constructs mutated or deleted for ATG(+1) were used, but alternative initiation at CTG(-315) or ATG(+418) did not lead to the expression of intranuclear forms. Residues 23 to 157 of the proregion were essential for export of the precursor, while mutation of the putative site for maturation by procollagen C-proteinase abolished processing to the mature form of the enzyme. Cross-linking of collagen, as measured by pyridinoline analysis, increased twofold with the recombinant cells, compared to non-transfected controls. This shows the specific contribution of LOX, as opposed to other genetic forms of the enzyme, to cross-linking in a cellular context.     

Model-based interpretation of cardiac beats by evolutionary algorithms: signal and model interaction

This paper presents a new approach for cardiac beat interpretation, based on a direct integration between a model and observed ECG signals. Physiological knowledge is represented by means of a semi-quantitative model of the cardiac electrical activity. The interpretation of cardiac beats is formalized as an optimization problem, by minimizing an error function defined between the model's output and the observations. Evolutionary algorithms (EAs) are used as the search technique in order to obtain the set of model parameters reproducing at best the observed phenomena. Examples of model adaptation to three different kinds of cardiac beats are presented. Preliminary results show the potentiality of this approach to reproduce and explain complex pathological disorders and to better localize their origin.     

Initiation of the breakage-fusion-bridge mechanism through common fragile site activation in human breast cancer cells: the model of PIP gene duplication from a break at FRA7I

Gene amplification plays a critical role in tumor progression. Hence, understanding the factors triggering this process in human cancers is an important concern. Unfortunately, the structures formed at early stages are usually unavailable for study, hampering the identification of the initiating events in tumors. Here, we show that the region containing the PIP gene, which is overexpressed in 80% of primary and metastatic breast cancers, is duplicated in the breast carcinoma cell line T47D. The two copies are organized as a large palindrome, lying 'in loco' on one chromosome 7. Such features constitute the landmark of the breakage-fusion-bridge (BFB) cycle mechanism. In hamster cells selected in vitro to resist cytotoxic drugs, common fragile site (CFS) activation has been shown to trigger this mechanism. Here, we characterize FRA7I at the molecular level and demonstrate that it lies 2 Mb telomeric to the PIP gene and sets the distal end of the repeated sequence. Moreover, our results suggest that the BFB process was frozen within the first cycle by healing of the broken chromosome. T47D cells thus offer a unique opportunity to observe the earliest products of the BFB cycle mechanism. Our findings constitute the first evidence that this amplification mechanism can be initiated in vivo by fragile site activation.