In vitro antimalarial activity of vegetal extracts used in West African traditional medicine.

Among strategies for the development of new antimalarials, a study of plants traditionally used in Africa against malaria has been pursued. Extracts obtained from the plants Azadirachta indica, Cinnamonum camphora, Lippia multiflora, Vernonia colorata, Guiera senegalensis, Combretum micranthum, and Ximenia americana, commonly used in Cote d'Ivoire by native healers for the treatment of malaria, were tested on two strains of Plasmodium falciparum: FcB1-Colombia (chloroquine-resistant) and F32-Tanzania (chloroquine-sensitive). Extracts were obtained after infusion and decoction, both techniques being used by most native healers. The antimalarial activities of the extracts were tested first by parasite 3H-hypoxanthine incorporation and second by visual evaluation of the activities of plant extracts on thin blood smears, which also permitted the determination of parasitic stages and parasite alteration. Among the seven plants tested, some had an apparent inhibitory effect on P. falciparum growth in vitro, while other seemed to be less efficient.

Ganciclovir resistance mutations in UL97 and UL54 genes of Human cytomegalovirus isolates resistant to ganciclovir.

Human cytomegalovirus (HCMV) resistance to ganciclovir results from mutations in viral phosphotransferase (UL97) and/or DNA polymerase (UL54) genes. The HCMV isolates from the blood of immunocompromised patients with persisting presence of the pp65 antigen in the blood in spite of ganciclovir therapy were tested for ganciclovir susceptibility by an immediate-early antigen plaque reduction assay, and the UL54 and UL97 genes were sequenced. Nine isolates from eight patients (six patients with acquired immune deficiency syndrome (AIDS), one liver transplant recipient and one renal transplant recipient) showed phenotypic resistance to ganciclovir. All these ganciclovir-resistant HCMV isolates harbored one or more of the following UL97 mutations: M460V, A594V, A594T, L595S, C603W, and M615V. Two isolates harbored the P522S mutation in the UL54 gene. The M615V mutation in the UL97 gene has not been reported earlier and its role in ganciclovir resistance remains to be elucidated. In ganciclovir-resistant HCMV isolates the UL54 gene was less frequently mutated than the UL97 gene. The P522S mutation was relatively frequent in UL54-mutated HCMV isolates.

[Various protocols for determining estrogens by the enzymatic method using estradiol dehydrogenase. Respective procedures and advantages]. / Les divers protocoles de dosage des estrogènes par la méthode enzymatique à l'estradiol déshydrogénase. Mode opératoire et avantages réciproques.

Up to now, more than 40.000 determinations of urinary estrogens (E1 + E2) have been carried out in routine clinical analysis by the enzymatic method using estradiol dehydrogenase. This method makes use of the transhydrogenating activity of the placental enzyme: this enzyme transfers hydrogen from NADP to NAD with recycling of the specific substrate (E1 + E2). For several years the necessary reagents have been commercially available in the form of a kit. Nonetheless, various improvements have been made to the measurement of reduced NAD, which accumulates in the reaction medium and is directly proportional to the concentration of the two estrogens. Three protocols are available at present: Spectrophotometric measurement at 340 nm (initial technique); Colorimetric measurement at 492 nm. The pink colour measured arises from the reduction of a tetrazolium salt (INT) by reduced NAD in a coupled system using diaphorase; Measurement by bioluminescence of the light energy liberated on the reduction of flavin derivatives by NADH. The reaction is mediated by various enzymes isolated from marine bacteria (FMN oxidoreductase and luciferase) in the presence of an aliphatic aldehyde (decanal). The procedure for each of these protocols is described as well as the means for controlling the linearity of the reaction. The choice of protocol is determined by the biological fluid available, the speed of response desired and the cost of the analysis.

[Antiphospholipid antibodies in African women presenting obstetrical complications]. / Anticorps antiphospholipides en gynécologie-obstétrique à Abidjan: intérêt diagnostique et pronostique.

The presence of antiphospholipid antibodies (APL) has been investigated in 68 African women presenting obstetrical complications versus a control group of 22 pregnant healthy women. Patients inclusion criterias were recurrent foetal loss, pre eclampsia, retroplacental haematoma and chronic foetal suffering. Eight of patients (11.8%) had a positive APL diagnosis, versus 0% in controls, using the combination of diluted thromboplastin test and Staclot LA These results argue for the systematic APL screening in African women with obstetrical complications, and could further improve the management of patients at risk.

Lipid, apolipoprotein AI and B levels in Ivorian patients with sickle cell anaemia.

Plasma apolipoprotein AI, apolipoprotein B, cholesterol and triglycerides in 114 Ivorian patients with homozygous sickle cell anaemia (HbSS) were compared to 40, age and sex matched controls with normal haemoglobin (HbAA). Except for plasma triglycerides, lipid and apolipoprotein levels were found to be lower in patients with sickle cell anaemia as compared to normal control subjects. The differences between the two groups were found to be statistically significant (p < 0.05 Student's t test). In addition, a highly significant decrease of plasma apolipoprotein AI (p < 0.01) was observed during painful crisis as compared to the steady state (0.78 g.L-1 versus 1.01 g.L-1), whereas triglyceride levels were found to be higher (1.02 g.L-1 versus 0.78 g.L-1). Although the metabolism of lipoproteins is being altered, the results of this cross-sectional study do not suggest that patients with sickle cell anaemia are at higher risk of developing coronary artery disease.

[Evaluation of atherogenic risk in homozygous sickle cell disease: study of lipid and apolipoprotein AI and B plasma levels]. / Evaluation du risque athérogène chez le drépanocytaire homozygote: étude des taux plasmatiques des lipides et apolipoprotéines AI et B.

The main clinical use of measurements of Apo Al and Apo B is to determine a patient's risk status for the development of ischemic heart disease. Apo B is generally accepted as a contributory cause of coronary artery disease, while Apo Al apparently has a protective effect. The present study reports the plasma change of Apo Al and B in sickle cell anaemic subjects. Immunochemical findings revealed that Apo Al and B levels were found to be lower in sickle cell patients as compared with normal subjects (HbAA). The atherogenicity index given by Apo B/Apo Al remained in the normal range during periods of steady state (0.64 +/- 0.28), while subjects in painful crisis had high index values (0.95 +/- 0.35). Therefore, we concluded that epidemiological studies on a large sample of patients are needed to confirm this relationship between painful crisis and risk of developing coronary artery disease.

[Enzymatic determination of urinary estrogens. A 5-year experience in a hospital milieu]. / Dosages enzymatiques des estrogènes urinaires. Une expèrience de cinq ans en milieu hospitalier.

The enzymatic method for urinary estrogens determination from Nicolas et al. has become a useful tool for the management of infertility problems. It can be used for: investigation of ovarian by establishing the urinary estrogens profile during menstrual cycle, useful to: understand anomalies of the spontaneous cycle, explain some therapeutic failures during IVF attempts or artificial inseminations (AID or AIC); prediction of failures during IVF attempts during spontaneous cycles, and monitoring ovarian response during stimulated cycles in order to determine the trigger with hCG; monitoring ovulation during induction of ovulation in anovulatory patients stimulated with various drugs (clomiphene citrate, pure FSH combination of FSH and LH, GnRH ou analogs...) under various conditions of prescription and administration (oral, IM, intermittent pulsatile administration with portable pump with or without hypophyseal down regulation). This technique allows also exploration of androgens after changing main androgens (delta 4 A, T, DHEA, DHEA S) into estrogens through the action of placental aromatase, as well as appreciation of aromatase activity of some tissues in the presence of androgenic substrates. This paper gives the conclusions after 5 years of practice with this method and summaries different works published by the biologists who developed the method and by the clinicians who used its results.

[Stimulation of ovulation by pure FSH for in vitro fertilization. Study of the route of administration]. / Stimulation de l'ovulation par la FSH pure en vue de fécondation in vitro. Etude de la voie d'administration.

Pulsatile injection of FSH to stimulate ovulation for in vitro fertilization was undertaken in order to compare the intramuscular route with the pulsatile subcutaneous route in two groups of 6 patients each selected at random. The patients were selected in such a way as to reduce as far as possible the parameters that would make it difficult to interpret the results. Both from the point of view of the numbers of patients who responded to stimulation of ovulation as well as the numbers of pregnancies that were obtained, the intramuscular route seems to be preferable.

Follicular steroids in relation to oocyte development and human ovarian stimulation protocols.

Oocytes of pre-ovulatory follicles were collected by laparoscopy for in-vitro fertilization in 118 women. Patients were treated either during the natural cycle, or after induction of ovulation with clomiphene citrate, or with human menopausal gonadotrophin (HMG), or clomiphene citrate combined with HMG. The oestrogens (oestrone + oestradiol) and total aromatizable androgens (androstenedione, testosterone and dehydroepiandrosterone and its sulphate) were assayed in follicular fluids using an enzymatic method. The levels of progesterone, 17 alpha OH-progesterone, thromboxane, and prostaglandins (PGE2 and PGF2 alpha) were determined by radioimmunoassay. The best follicular fluid indicator of oocyte fertilization in vitro was the A/E ratio, which was less than 1 when the oocyte was fertilized in vitro and led to a pregnancy. The lowest value for the A/E ratio was obtained with spontaneous ovulation protocols. Regardless of oocyte development, the progesterone level was always greater than 2000 ng/ml, and the PGE2/PGF2 alpha ratio was greater than 1 in all stimulated cycles. Our investigations show that a combination of clomiphene citrate and HMG provides the best stimulation, on the basis of follicular fluid analysis and the outcome of fertilized oocytes.

[Pregnancies by fertilization in vitro. Biochemistry of the follicular fluid]. / Grossesses par fécondation in vitro. Biochimie du liquide folliculaire.

Fifty four follicular fluids containing 41 oocytes were sampled from 6 patients in which at least one pregnancy was obtained after IVF. Their biochemical composition has been correlated with the follicle maturity and oocyte quality. Three components have been assayed: steroids, prostanoids, proteolytic enzymes. Determinations of estrogens and total aromatisable androgens were carried out by the enzymatic technic of Nicolas and al.; progesterone and 17 OH progesterone by RIA. Development of in vitro cultured and fertilized oocytes appears to be improved when progesterone and estrogens concentrations in the follicular fluid are above 2 000 and 600 ng/ml respectively, with an A/E ratio under 1. On the other hand, prostanoids (T X B2, PGE2, PGF2 alpha determined by RIA) and proteolytic enzymes : (collagenolytic and kallikrein type activities) seemed related to ovulatory activity rather than to oocyte quality.