Avoidance, biomass and survival response of soil dwelling (endogeic) earthworms to OECD artificial soil: potential implications for earthworm ecotoxicology.

Soil dwelling earthworms are now adopted more widely in ecotoxicology, so it is vital to establish if standardised test parameters remain applicable. The main aim of this study was to determine the influence of OECD artificial soil on selected soil-dwelling, endogeic earthworm species. In an initial experiment, biomass change in mature Allolobophora chlorotica was recorded in Standard OECD Artificial Soil (AS) and also in Kettering Loam (KL). In a second experiment, avoidance behaviour was recorded in a linear gradient with varying proportions of AS and KL (100% AS, 75% AS + 25% KL, 50% KS + 50% KL, 25% AS + 75% KL, 100% KL) with either A. chlorotica or Octolasion cyaneum. Results showed a significant decrease in A. chlorotica biomass in AS relative to KL, and in the linear gradient, both earthworm species preferentially occupied sections containing higher proportions of KL over AS. Soil texture and specifically % composition and particle size of sand are proposed as key factors that influenced observed results. This research suggests that more suitable substrates are required for ecotoxicology tests with soil dwelling earthworms.

Effects of silver nanoparticles on survival, biomass change and avoidance behaviour of the endogeic earthworm Allolobophora chlorotica.

Increasing commercial application of silver nanoparticles (Ag NP) and subsequent presence in wastewater and sewage sludge has raised concerns regarding their effects in the aquatic and terrestrial environment. Several studies have employed standardised acute and chronic earthworm-based tests to establish the toxicological effects of Ag NP within soil. These studies have relied heavily on the use of epigiec earthworm species which may have limited ecological relevance in mineral soil. This study assessed the influence of Ag NP (uncoated 80nm powder) and AgNO3 on survival, change in biomass and avoidance behaviour in a soil dwelling (endogiec) species, Allolobophora chlorotica. Earthworms were exposed for 14 days to soils spiked with Ag NP or AgNO3 at 0, 12.5, 25, 50 and 100mgkg-1 either separately for survival and biomass measurement, or combined within a linear gradient to assess avoidance. Avoidance behaviour was shown to provide the most sensitive endpoint with an observable effect at an Ag NP/AgNO3 concentration of 12.5mgkg-1 compared with 50mgkg-1 for biomass change and 100mgkg-1 for survival. Greater mortality was observed in AgNO3 (66.7%) compared with Ag NP-spiked soils (12.5%) at 100mgkg-1, attributed to increased presence of silver ions. Although comparison of results with studies employing Eisenia fetida and Eisenia andrei suggest that the A. chlorotica response to Ag NP is more sensitive, further research employing both epigeic and endogeic earthworms under similar experimental conditions is required to confirm this observation.

Role of physical activity and sport in oncology: scientific commission of the National Federation Sport and Cancer CAMI.

This overview reports published data about the interaction between physical activity and sport during and after cancer on one hand and improvement in psychological parameters, survival and biological mechanisms underlying this effect on the other hand. Practising physical activity and sport during cancer modifies parameters assessing fatigue and quality of life and reduces symptoms of depression. An association also exists between the practise of physical activity and sport and overall and cancer-specific survivals, especially after breast cancer, colon cancer and prostate cancer. These benefits seem to be mediated by a modification of circulating levels of estrogens, insulin, IGF-1 and by a decrease in insulin-resistance, by alterations in the secretion of adipokines, and by a reduction in chronic inflammation through decreased levels of cytokines. There exist some obstacles to the practise of physical activity. These obstacles are mainly related to a fear of pain induced by physical activity and to overweight. These programmes of physical activity and sport cannot be offered to all patients since there are several contra-indications, with some being present since the initial visit and others appearing during cancer management either due to disease progression or related to iatrogenic effects. Whereas benefits from physical activity and sport among cancer patients seem obvious, there are still several pending clinical and biological issues.

Dietary enrichment with alpha-linolenic acid during pregnancy attenuates insulin resistance in adult offspring in mice.

OBJECTIVE: Our objective was to test the contribution of dietary enrichment in essential or saturated fatty acids, in normocaloric diets, on the lipid accumulation and insulin resistance in the adult offspring in a C57Bl6/J mice model. METHODS: Pregnant mothers were fed normocaloric diets containing 6% fat enriched in essential fatty acids (EFA): alpha-linolenic (ALA-18:3, n-3), linoleic (LA-18:2, n-6), or saturated fatty acids (SFA). After a washing-out period with regular diet, the offspring received a high-fat diet before euthanization. RESULTS: Adult mice fed maternal ALA showed lower body weight gain and lower liver fat accumulation, lower HOMA index and lower stearoyl-CoA desaturase (SCD1) activity than those fed maternal SFA. CONCLUSION: The results observed using this novel model suggest that ALA in maternal diet may have the potential to inhibit insulin resistance in adult offspring.

Photodynamic ablation of a selected rat embryo: a model for the treatment of extrauterine pregnancy.

BACKGROUND: To test the feasibility of photodynamic therapy (PDT)-based ablation of rat embryos as a model for PDT of extrauterine pregnancy (EUP) in humans. METHODS: A controlled pre-clinical study. Selected rat embryos [one per litter, n = 30, embryonic day 14 (E14)] were subjected to placental injection of a Palladium-bacteriochlorophyll derivative and illuminated to achieve selective photo-ablation. Histopathology studies were performed 48 h after treatment (E16). Parturition (E21) and breeding (approximately 12 weeks) after treatment were also evaluated. RESULTS: Using direct placental injection, nearly 80% of the treated rat embryos were selectively photo-ablated, leaving the remaining litter unharmed to achieve normal parturition. Treated animals retained fertility and normally implanted in both treated and untreated uterine horns attesting to the confined toxicity inherent to this approach. CONCLUSIONS: Although requiring respective adaptation to clinical application in terms of treatment protocols and designated hardware, photodynamic interventions using novel bacteriochlorophyll-based photosensitizers may prove applicable to treatment of EUP, as well as other gynecological pathologies and malignancies in a safe, minimally invasive manner.

[What is the validity limit of patient reimbursement? Is age a criterion for refusal? No...]. / Au-delà de quelle limite votre ticket n'est plus valable? L'âge est-il un critère de refus? Non...

The late 20th century trend to delay birth of the first child until the age at which female fecundity or reproductive capacity is lower has increased the incidence of age-related infertility. In the female, the number of oocytes decreases with age until the menopause. Oocyte quality also diminishes, due in part to increased aneuploidy. A significantly lower pregnancy rate per stimulation and delivery rate per retrieval is found in women aged 40 to 45 years when compared with women aged 25 to 39 years. However, acceptable pregnancy can be achieved in woman aged 40 to 43 years (4 to 8%). Furthermore, very few women aged over 40 ask for IVF procedure.

Intracytoplasmic sperm injection in infertile patients with structural chromosome abnormalities.

In the present study we investigated the results of cytogenetic analysis in male and female patients included in an intracytoplasmic sperm injection (ICSI) programme for severe male infertility as well as in conceptuses resulting from these ICSI treatments. In the 261 couples treated, 11 male (4.2%) and three female (1.2%) abnormal karyotypes were found, all consisting of structural chromosome anomalies. Chromosomal translocation exhibited the highest frequency (eight males and two females), and there were also three cases of chromosomal inversion (two males and one female) and one male with one additional marker chromosome. There was no difference in fertilization rates among couples with abnormal (n = 14) and normal (n = 147) cytogenetic results, and the rates of clinical pregnancy per ICSI attempt were 25.0% (5/20) and 20.6% (78/ 378) respectively. In pregnancies obtained in couples with normal karyotypes, all of the 108 fetuses were free of chromosomal abnormalities. Among the eight fetuses from couples with chromosome structural anomalies, three out of five and two out of three inherited from the cytogenetic defects found in their father or mother respectively. In this series of 83ICSI pregnancies there were no chromosomal abnormalities other than those inherited from the parents. These findings suggest that normal pregnancy rates can be obtained by ICSI in cases of chromosomal translocation in couples with severe male infertility. However, until further evaluations of available data can be performed, cytogenetic analysis must be conducted prior to ICSI in men with low sperm counts, and genetic counselling must include prenatal diagnosis for all growing conceptuses.

Spermatid injection into human oocytes. II. Clinical application in the treatment of infertility due to non-obstructive azoospermia.

We have reported recently the first birth after intrauterine transfer of embryos obtained by injection of round spermatids into oocytes in cases of unexpected azoospermia. Here we provide a complete documentation of the series of 11 cases in which this novel method of infertility treatment was employed. In four of these cases, elongated spermatids were identified in the ejaculate, and it was decided to perform elongated spermatid injection (ELSI). In the other six cases, only round spermatids were present, and round spermatid injection (ROSI) was done. In one case, ROSI was given preference to ELSI because of a very poor viability status of elongated spermatids present in the ejaculate. Fertilization of at least one oocyte was achieved in 10 of the 11 treatment cycles; the fertilization rate in these 10 cycles ranged between 7 and 100% with a mean value of 45%. All of the two-pronucleated zygotes cleaved and were transferred to the patient's uterus. A singleton pregnancy was achieved in two ROSI cycles. Both pregnancies developed uneventfully and resulted in the birth of normal infants. These data show the intra-ooplasmic injection of spermatids obtained from the ejaculate may become the treatment of first choice in patients with non-obstructive azoospermia.

Immunoregulatory activity in supernatants from cultures of normal human trophoblast cells of the first trimester.

Supernatants from isolated trophoblast cell cultures (trophoblastic fluid) derived from first-trimester human placentas were assessed for immunoregulatory activity. Trophoblastic fluid at different days of culture consistently inhibited the blast transformation of allogenic lymphocytes. This suppressor effect had no apparent correlation with biosynthesis of human chorionic gonadotropin by trophoblast cells, since this hormone was secreted into the culture fluid only for the initial 3 days. However, the culture fluids of such purified trophoblast cells contained an immunosuppressive factor, pregnancy-associated plasma protein A, which was measurable throughout the culture period of 8 days. The presence of pregnancy-associated plasma protein A in significant amounts in trophoblastic fluid collected at daily intervals indicated a continuous secretion ability of pregnancy-associated plasma protein A by trophoblast cells in culture parallel to the suppressive immunoregulatory effect of the fluid. Such immunosuppressive effect was absent in the culture fluids of control BeWo malignant trophoblast cells; the BeWo cell culture fluids had markedly reduced levels of pregnancy-associated plasma protein A. The culture supernatant of normal trophoblast cells of placentas from first-trimester pregnancy activated in vitro the generation of a population of suppressor lymphocytes. This effect is generally considered responsible for immunologic tolerance. Therefore demonstration of immunosuppressive effects and the presence of relatively high levels of pregnancy-associated plasma protein A in trophoblastic fluid indicate that such proteins secreted by the trophoblast cells may be important in the local immunoregulatory processes of the fetal allograft.

Ovarian stimulation for in-vitro fertilization combining administration of gonadotrophins and blockade of the pituitary with D-Trp6-LHRH microcapsules: pilot studies with two protocols.

In women undergoing in-vitro fertilization and embryo transfer (IVF-ET), a total of 408 IVF cycles were stimulated using human menopausal gonadotrophin (HMG) or pure follicle stimulating hormone (FSH) plus HMG in combination with a single injection of D-Trp6-LHRH microcapsules in order to enhance the ovarian response to gonadotrophins and to avoid spontaneous LH surges. Sixty-seven pregnancies were achieved. Two protocols were employed. In protocol 1 ('blocking protocol', n = 268), the pituitary was first inhibited with a full dose (3.75 mg) of D-Trp6-LHRH in microcapsules and ovarian stimulation was started after the hypogonadotrophic hypogonadal state was ascertained (E2 less than 50 pg/ml). In protocol 2 ('flare-up protocol', n = 140), the treatment with D-Trp6-LHRH microcapsules (half-dose = 1.80 mg) and the ovarian stimulation with gonadotrophins were started at the same time. Higher doses of gonadotrophins were needed (39.5 +/- 11.2 ampoules FSH and/or HMG) in protocol 1, in which the pituitary was blocked prior to and during the stimulation, than in protocol 2 (20 +/- 9 ampoules) where the exogenous gonadotrophin stimulation appeared to be augmented by the initial agonistic effect of the injection of D-Trp6-LHRH microcapsules. In patients with purely tubal infertility, under 38 years old and no male factor, the results obtained with protocols 1 and 2 were similar in terms of pregnancy rate per cycle or per embryo transfer: 22.6 versus 20.5% and 28.3 versus 27.4%, respectively. However, considering the cost benefit, 'flare-up' protocols appeared to be a better choice and could be recommended.

In vitro production of pregnancy-associated plasma protein-A (PAPP-A) by trophoblastic cells.

Pregnancy-associated plasma protein-A (PAPP-A) a high molecular weight glycoprotein, is found in high concentration in the maternal circulation during pregnancy. Immunohistochemically, PAPP-A can be localized in the trophoblast and in the decidua. Short term cultures of trophoblastic and decidual explants produce PAPP-A in vitro. It was thus of interest to see if long term cultures of primary monolayers of human trophoblast cells were capable of producing PAPP-A. Under our in vitro conditions, trophoblastic monolayers were producing both PAPP-A and beta hCG. During the first 3 days of culture PAPP-A levels increased in the medium whereas beta hCG levels decreased. The production of both proteins could be inhibited by cycloheximide. These results strongly suggest that the trophoblast is a source of PAPP-A in vivo.

The effects of indomethacin administration during pregnancy on women's and newborns' T-suppressor lymphocyte activity and on HLA class II expression by newborns' leukocytes.

It has been previously shown that T lymphocytes from human newborns and pregnant women exert a suppressive activity when assayed on the PWM-induced B cell maturation. The mechanisms of the suppression have remained entirely unknown. Prostaglandin E2, known to trigger T-cell mediated suppressive activity, may be involved. We took advantage of the treatment of pregnant women with indomethacin, because of premature labor or hydramnios, to investigate the role of prostaglandins in the activation of T suppressor (TS) activity. Administration of indomethacin (250 mg/day for 1-7 weeks, then 150 mg/day for 3-12 weeks) during the third trimester of pregnancy, abrogated the TS activity in the nine women and the three newborns tested. Abrogation of TS activity by indomethacin therapy led to normal PWM-induced B cell maturation in pregnant women but not in newborns. Moreover, the low expression of HLA class II antigens observed on normal newborn B lymphocytes and monocytes was corrected in newborns from indomethacin-treated mothers. Our results strongly suggest that prostaglandins may play a role in induction of TS activity observed in normal pregnant women and newborns and in the decreased expression of HLA class II antigens on newborns' leucocytes. Both phenomena could play a role in immunological interactions between mother and fetus.

[Immunoregulatory aspects of placental function]. / Aspects immuno-régulateurs de la fonction placentaire.

An immunological response from mother to foetus is known to exist during pregnancy. Multiple mechanism ensure that this response is not one of rejection but of "tolerance" of the foetus, and the placenta, as the site of maternal-foetal exchanges, plays a prominent role in this respect: (1) the placenta cell bears foetal antigens, half of which are of paternal origin; it constitutes therefore the antigenic stimulus of maternal immune response; (2) this response can be modulated quantitatively through placental hormones and glycoproteins - an action which has been demonstrated in vitro but which may well take place at utero-placental level; (3) finally, the placenta can act qualitatively on the maternal response itself by reducing cytotoxic reactions responsible for graft rejection and by stimulating immunosuppressor cell populations. The therapeutic applications of some of these properties of the placenta are being investigated in the fields of rheumatology and renal transplantation. They will be considered in obstetrics only when more precise data are available concerning the immunopathological mechanisms involved in spontaneous abortion, preeclampsia and foetal growth retardation.

HLA-DR antigen on human trophoblast.

To assess the presence of human leukocyte antigens (HLA) on first trimester human trophoblast cells, frozen sections of villous trophoblast and monolayer cultures of isolated cells from placental villi were prepared and exposed to a mouse monoclonal antibody directed against HLA-DR and then incubated with fluorescein-conjugated goat antimouse antibodies. Fluorescence microscopy demonstrated that HLA-DR antigens were present on only the small polygonal epithelioid cells of the monolayer culture. The crescentic staining pattern was consistent with widespread distribution of antigen on the cell membrane. There was no staining over giant multinucleated structures or on fibroblasts of such cell cultures. No HLA-DR was detected when this indirect immunofluorescent technique was applied to tissue sections of villous trophoblast. Existence of high concentrations of hCG in culture supernatants and coincident localization of both hCG and HLA-DR using antibodies conjugated with rhodamine or fluorescein on the polygonal epithelioid cells indicate the trophoblastic origin and expression of HLA-DR antigen under in vitro monolayer culture conditions.

[Considerations after 10 years of treating leg ulcers]. / Réflexions sur dix ans de traitement des ulcères de jambe.

There are four times as many varicose ulcers as arterial ulcers, this being basically the case of patients over 50. Venous thrombosis is responsible for about one-third of the varicose ulcers, and atherosis causes more than one-third of the arterial ulcers. The latter often involve metabolic disorders: this is not the case with varicose ulcers. When there is superinfection, pyocyanic is rampant and is followed by yellow staphylococci. Arterial ulcers are very often painful and cicatrisation is slow, often even hopeless, whereas varicose ulcers heal when the responsible varices are cured.