Distribution et activités des alcaloïdes défensifs des Coccinellidae

From a survey of 30 species and varieties of ladybugs the presence of alkaloids appears to be correlated with the existence of aposematic colour and not with being carnivorous or phytophagous. The alkaloids described until now all belong to the Coccinellini and are closely related, but other types of bases have been detected in some genera. The observed distributions are in agreement with the modern taxonomy of the family.Ladybug alkaloids constitute an effective defence against ants, Myrmica rubra, and quails, Coturnix coturnix, but all the beetles containing alkaloids do not possess the same degree of protection. Individual quail react differently towards moderately protected species.The bioassay used for the first isolation of coccinelin is described. The repulsive activities of aqueous solutions of coccinellin and convergin towards ants have been compared.     

Spermatozoïdes,environnement et prévention

Over the last fivty years, the risk factors for infertility have increased substantially, particularly those due to the environment. Spermatogenesis and spermatozoa can be affected by physical (ionizing radiation microwaves, heat, cryopreservation) or chemical agents (antimitotics drugs, antibiotics, tranquillizers, insecticides, pesticides, industrial solvants, some heavy metals, alcohol, cannabis etc.). Some natural factors, as stress or paternal age (ageing or very youthful age relatively to about thirty) also seems to affect spermatogenesis and, particularly, the age can be joined with the previous ones. On the whole, these factors are able to decrease the male fertility through some changes about the concentration, the motility or the morphology of spermatozoa and so it is possible to describe populations subject to the risk. Moreover, these spermatogenetic changes can lead abnormalities in progeny. For instance, some antimitotic drugs as cyclophosphamide, when administrated to the male rat, lead malformations or functional anomalies as behavioral troubles. The industrial solvents lead a decrease of the birth weight and the cannabis leads an increase of the ante-or post-natal death. Moreover, the change of the paternal spermatogenesis caused by cannabis can be found again in the male progeny. The problem is similar with the lead, the benzodiazepines and the alcohol. Concerning the physical factors, some authors have shown that the children born from radiation exposed fathers presented an increase of the probability of leukemia. In animal, the postimplantation loss is increased when the father is irradiated or subjected to heat before mating. Finally, the paternal ageing is responsible for new dominant autosomic mutations. Moreover, in animal and man, paternal ageing and, in man, very youthful age, also seems responsible for a gradual lowering in the level of progency cerebral functions. On the whole, these data should lead to an preventive attitude which would be more effective before about thirty years of age than after this period.     

Données récentes sur la maturation des spermatozoïdes dans l'épididyme humain

Au cours de leur transit épididymaire, les spermatozo?des rencontrent un environnement varié, au regard des protéines avec lesquelles ils rentrent en contract. Dans la partie proximale de l'épididyme, ils sont soumis à l'action d'enzymes, et exposés à des protéines susceptibles de modifier les membranes. Dans la partie moyenne prédomine une autre catégorie de protéines et d'enzymes; celles associées au transport de stérols pourraient modifier les membranes spermatiques afin de permettre l'ancrage de protéines de liaison à la zone pellucide: P 34H et CD52. Dans la partie distale les spermatozo?des rencontrent des activités croissantes d'enzymes lytiques, des protéines impliquées à la fois dans la liaison à la zone pellucidé et la fusion ovocytaire, l'antigène de maturation CD 52, une activité anti-microbienne, et enfin des agents décapacitants qui facilitent leur survie avant l'éjaculation. L'adhérence des protéines aux différents domaines membranaires (comme la région antérieure ou le segment équatorial de l'acrosome) peut dépendre de la nature de la protéine, de la composition lipidique de la région membranaire concernée, et de l'environnement ionique dans la lumière épididymaire. La localisation des protéines sur le spermatozo?de, qu'il soit à acrosome intact (membrane acrosomique) ou acrosome-réagi (segment équatorial) pourra dicter leur r?le, concernant par exemple la fixation à la zona (P 34H), ou la liaison à l'ovocyte (gp 20). Les protéines comme les membranes peuvent être modifiées durant le transit épididymaire par des enzymes qui peuvent ajouter ou retirer des sucres et des peptides de la surface spermatique.     

Anticorps anti-spermatozoïdes: techniques de dépistage et interprétation des résultats

Since the first publication on the detection of sperm-agglutinating antibodies in infertile men, multiple assays have been described. The most useful tests are able to detect antibodies bound to the sperm membrane of motile spermatozoa. The immunobeads test (IBT) is considered to be the most advantageous in terms of its sensitivity, the low incidence of false-positive results, and its ability to localize antibodies of different immunoglobulin classes on the sperm surface. The IBT assay can be used in parallel with the mixed antiglobulin reaction (MAR), to detect sperm-associated antibodies in the ejaculates of infertile men, the most rational way to test for antisperm antibodies (ASA) in males. In view of the high level of agreement between the two assays, MAR, the easier of the two, may be used as a first step in the detection of these antibodies. A positive MAR must be confirmed by IBT, as this assay is more specific for the detection of IgA antibodies. The clinical significance of sperm-associated antibodies is usually established according to the proportion of motile spermatozoa coated with immunobeads, its class and its localization on the sperm surface. However, binding of immunobeads does not provide any information about the antigens against which the antibodies are directed. As the functional effects of sperm-associated antibodies may vary as a function of their antigenic specificities, other assays, using purified fertilization related antigens, are necessary to establish, for each individual, the specific impact of the antibodies on the fertilization process. The indirect IBT assay has recently become the most widely used test to detect the various classes of ASA in serum and cervical mucus of infertile women and in the serum and seminal fluid of infertile men, in combination with the direct assay described above. However, in most laboratories, it is performed with only one dilution of the biological fluid tested, usually a low dilution, so that antibody levels of no significance for fertility could be detected. This may explain a recente debate (Human Reprod, 1999) on the significance of ASA as a cause of infertility. At present, and in the absence of standardized assays able to identify the antigens involved in each individual immune reaction, antibody assays, as detected by the IBT assay, in the serum and/or genital secretions of infertile subjects might provide useful clinical guidance.     

Morphologie des spermatozoïdes

Spermatozoa morphology is one of the qualitative characteristics of spermatogenesis. However, because of both the variations in the definition of normal morphology and the existence of different kinds of sperm abnormalities as well as the use of various techniques of morphology assessment, such a parameter is poorly used in usual laboratory work. Morphological sperm anomalies can be from testicular or post-testicular origines, while the latter is still unproved. The causes of such anomalies are either from genetic origines, but in these cases any spermatozoa demonstrate this anomaly, or due to an endogenous factor with varicocele the most usually quoted but unproved pathology, But exogenous factors, either chemical such as drugs and pesticides or physical such as heat, are also responsible for morphological sperm anomalies. Analysis of sperm morphology is indicative of both the testicular health status (in cases of occupational exposure to chemical or physical toxics) and the fertility potential since morphology is correlated to sperm motility and involved in fertilization through the acrosome reaction.     

Principes de préparation et de congélation des spermatozoïdes testiculaires humains

The advantages and feasibility of human testicular spermatozoa cryoconservation for intracytoplasmic sperm injection (ICSI) have now been clearly demonstrated. However, the freezing protocol is based on empirical knowledge obtained from freezing of ejaculated spermatozoa. Testicular spermatozoa may not be fully mature gametes and may also be retrieved in only limited quantities. Little research has been conducted to determine whether they have the same cryobiological requirements as ejaculated spermatozoa. A better understanding of their cryobiological features and assessment of possible subcellular changes after thawing would help to optimize testicular preparations for cryopreservation (whole biopsies, seminiferous tubules, shredded suspension, single spermatozoa, etc.), freezing-thawing procedure, freezing media, and storage. Finally, there is a growing need for welldefined criteria (nuclear quality, etc.) to evaluate the tolerance of testicular spermatozoa to freezing-thawing procedure for ICSI     

Tests de Selection des Spermatozoïdes avant Assistance Médicale à la Procréation

There are many procedures to select spermatozoa but those procedures are often specific of each andrology centre. This review that is not an exhaustive review of the numerous methods published is rather devoided to focus on the main techniques, the sperm washing, the filtration methods («SpermPrep»), the swim-up, and the discontinuous density gradients with colloïdal solutions («Percoll», PureSperm” and «Isolate») including iodinated organic molecule as ‘OptiPrep». The first part presents a synthesis of literature in order to bring out standardized procedures, and a summary of the advantages and disadvantages of each step of these procedures. Some specific anomalies of sperm need particular treatments. There is a consensus on the validated techniques to prepare semen samples in the presence of antisperm antibodies, leukocytes, bacterial contaminants, and also semen samples exhibiting hyperviscosity or in case of retrograde ejaculation. Cryopreserved sperm preparation is also described with references to experimental results in freezing-thawing human spermatozoa. The second part proposes guidelines for the therapeutic choice of the appropriate assisted reproductive technologies (i.e.: intra uterine artificial insemination, in vitro fertilization or intracytoplasmic sperm injection) according to semen parameters, and to the outcome of sperm selection and survival.     

Evaluation morphologique des spermatozoïdes

Semen analysis, as defined by World Health Organisation (WHO), is a fundamental step in the work-up of infertile couples. Spermatozoa morphology has been recognised as the best predictive factor in natural fertility, and in intrauterine insemination and classicalin vitro fertilisation. Ultrastructural spermatozoa abnormalities are the only sperm alterations likely to influence the outcome of ICSI. However, these abnormalities cannot be detected by conventional microscopy (¥100 or ¥200–400). Bartoov et al. (2002) developed a real-time spermatozoa observation system using a ¥6600 magnification called MSOME (Magnification Motile Sperm Morphology Examination). Spermatozoa abnormalities detected with this technique are vacuoles localised on spermatozoa heads with variable number, size and site (nucleus or acrosome). Spermatozoa evaluation using MSOME could be performed to predict the probability of fertilisation by these spermatozoa either spontaneously or after assisted reproductive technology.     

Liste des cyclopoïdes gnathostomes (Crustacés copépodes) des îles Britanniques et de l'Irlande

Résumé La liste de Cyclopoïdes gnathostomes donnée comprend 58 formes pour la Grande-Bretagne (Angleterre 56, Ecosse 40) et 38 pour l'Irlande.A titre de comparaison on peut mentionner qu'on en connaît actuellement 56 en France et 37 dans les Pays-Bas.Quelques remarques ont été faites sur des questions de nomenclature et concernant certaines espèces.     

Les corps myeloïdes de l'epithelium pigmentaire rétinien

Résumé Les corps myéloïdes de l'épithélium pigmentaire rétinien ont été observés après différentes techniques de fixation, déshydratation, inclusion et coloration. Après fixation classique par le glutaraldéhyde et l'osmium, l'espace intersacculaire apparaît réduit (jonctions) et une ligne dense est parfois observable dans l'espace intrasacculaire. La fixation par le permanganate met en évidence de façon permanente cette ligne dense qui correspondrait à la fusion des membranes. La coloration négative caractérise un constituant protéique entre les saccules. La déshydratation par le Durcupan hydrosoluble met en évidence un matériel lipidique intrasacculaire qui est extrait par le méthanol-chloroforme et partiellement par la déshydratation alcoolique classique.Les corps myéloïdes apparaissent plusieurs jours avant l'éclosion chez l'oiseau. Ils se forment à partir du réticulum granulaire. Ils ne peuvent être confondus avec les dictyosomes Golgiens : Ils ne contiennent pas de glycoprotéines.

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Summary Myeloid bodies of the retinal pigment epithelium of different vertebrates were investigated by various techniques of fixation, dehydration, embedding and staining. After classical fixation with buffered glutaraldehyde and osmium, the intersaccular space is narrow (junction complexes) and a dense line is sometimes visible in the middle of the intrasaccular space. With permanganate fixative this dense line is always visible; it represents probably the fusion of the unit-membranes. The negative staining demonstrates the presence of proteins between the saccules. Dehydration by water soluble Durcupan shows a lipidic component in the intrasaccular space, which is removed by methanol-chloroform and partly by the classical ethanol dehydration.The existence of myeloid bodies is demonstrated in the bird-embryo a few days before birth. They originate from granular reticulum cisternae. They are not of Golgi origin: glycoprotein cannot been demonstrated at their level.

Chargé de Recherche à l'I.N.S.E.R.M. (contrat n° 711.152). Avec la collaboration technique de L. Salin et D. Joseph.     

Anticorps anti-spermatozoïdes: Revue de la littérature

Auto immune infertility represents about 15% of all sterilities. The most commun mechanisms of autoimmunity in males are genitals infections (epidymitis) and traumatisms (vasectomies, testiculat biopsies). Mixt agglutination reaction test and immunobead test are the most commun tests used for detection of antibodies. Since 1981, researchs tried to characterize spermatozoa antigens by using polyclonal and monoclonal antibodies. More than 30 antigens had been found during this last decade. Naz and coll had identified an antigen called FA 1 antigen. FA 1 Ag is a 23 Kd glycoprotein. Anti FA1 antibodies inhibit binding and penetration of sperm to zona intact mice oocytes and also zona free hamster oocytes. They do so without causing agglutination or immobilization of sperm and by the way they cannot be detected by usual tests.     

Les spermatozoïdes immobiles frais ou décongelés en fécondation assistée

Contrasting with sperm count or morphology, complete lack of mobile sperm may seriously impair ICSI fertilization and pergnancy rate. In three cases with flagellar skeleton abnormalities [dynein arm absence] only immobile sperm were found in the ejaculate. Following repeated ejaculations, higher rates of viable spermatozoa and even some motile spermatozoa could be found. Some times, in nonobstructive azoospermia, extensive sperm search didn't allow us to find but immobile sperm mostly, with very few motile sperm cells, not enough for the microinjection of all oocytes. The third group of immobile sperm is iatrogenic, following freezing and thawing surgically retrieved, testicular or epididymal spermatozoa in order to avoid repeated surgical retrieval. Following thawing, one find frequently very few motile spermatozoa that may be not enough for all retrieved oocytes and it might be necessary to inject some eggs with immobile spermatozoa. The outcome of ICSI using mobile and immobile sperm was compared in the three above mentioned groups: 1-immobile ejaculated sperm with flagellar defects, 2-immobile sperm discovered in the ejaculate after extensive sperm search and 3- immobile frozen-thawed testicular or epididymal spermatozoa. The results of ICSI in these groups show that fertilizing ability of fresh or frozenthawed immobile spermatozoa is not significantly different from ICSI with mobile sperm from the same origin. More over, in the first group with flagellar abnormalities, repeated ejaculations allowed us significantly increase sperm viability and fertilization ability. Finding only immobile fresh or frozen-thawed sperm the day of egg retrieval should not lead us to ICSI cancellation. Pregnancies may occur with such immobile sperm.     

Estimation du taux d’aneuploïdies des spermatozoïdes épididymaires prélevés en vue d’ICSI chez des hommes à caryotype normal

Over the last ten years, fluorescent in situ hybridization in decondensed sperm nuclei has been used to study the chromosomal constitution of human spermatozoa. Studies have estimated that the disomy rate per chromosomal pairs is between 0.15% and 0.3%. The aim of this study was to evaluate the aneuploidy rate of human epididymal spermatozoa extracted from five men with obstructive azoospermia undergoing IVF. Genetic studies (karyotypes, Y micodeletion syndrome and mutation of the CFTR gene) did not reveal any abnormality. Disomy frequencies were determined by X-Y-8 multicolour fluorescence in situ hybridisation on 18,013 epididymal spermatozoa and 20,000 spermatozoa from healthy donors (control group). No significant difference was found between epididymal and ejaculated samples. However, isolated non-significant differences were observed between one of the patients and the control group. In conclusion, the present findings suggests that there is no increased risk for de novo chromosomal aberrations after IVF therapy with epididymal spermatozoa of men with obstructive azoospermia.     

La vitalité des spermatozoïdes

Most of the numerous techniques used to assess sperm viability only have research applications, while only two classical tests, i.e. eosin-Y and hypo-osmotic swelling test (HOST), are currently used in routine sperm analysis to determine the percentage of viable sperm. A viability rate below 50% of living sperm defines necrozoospermia, a condition whose clinical significance is fairly difficult to assess as the mechanisms of sperm cell death are still poorly understood. However, even when a precise cause for necrozoospermia cannot be identified, abnormal viability requires further andrological investigations with particular emphasis on clinical and laboratory signs of chronic infection of the male reproductive tract. Intracytoplasmic sperm injection (ICSI) can yield very good pregnancy rates, even in couples with the most severe forms of male infertility. However, when no motile sperm are available after sperm preparation, the outcome of ICSI is seriously impaired, probably because of a high risk of injecting dead sperm. In these patients, sperm viability could therefore be assessed by the hypo-osmotic swelling test in order to select only viable sperm for ICSI. However, the long incubation time of sperm in the hypo-osmotic solution, as recommended in the classical HOST procedure, has been shown to be detrimental to the spermatozoa. A single sperm test able to assess the viability of each individual spermatozoon within microdroplets covered by mineral oil therefore seems to be preferable. This selection procedure is less suitable in the case of immotile frozen-thawed sperm, as viability does not appear to be reliably predicted by HOST in cryopreserved sperm. Examination of sperm viability now also evaluates programmed cell death or apoptosis, as apoptotic alterations can be detected in spermatozoa by several techniques. The percentage of apoptotic sperm is correlated with deficient sperm parameters and poor outcome of assisted reproductive techniques. More effective selection procedures are therefore needed in order to identify spermatozoa not only with intact membranes but also with an intact genome to be used for ICSI.     

Chimiotaxonomie des lycopodiales: distribution des alcaloïdes

The distribution of lysine-derived alkaloids in the genus Lycopodium (s.l.) supports the separation of the following taxa, Huperzia (= Urostachys), Lycopodiella s.l. (= Lepidotis, excl. L. volubile and L. deuterodensum) from Lycopodium s. str. Within the latter, the Fastigiatum group and the Complanata section (= Diphasiastrum) can be distinguished. The results obtained are in good agreement with the classification proposed by Wilce.     

Hyperparasitisme facultatif de parasitoïdes en cours de développement par les femelles des ectoparasitoïdes Eupelmus vuilleti et Eupelmus orientalis Craw

In a community of three ectoparasitoids, Dinarmus basalis, Eupelmus vuilleti and E. orientalis, the host Callosobruchus maculatus parasitised 48 h before by D. basalis, is accepted by E. vuilleti females after they have eliminated the eggs and neonatal larvae of D. basalis. This ovicidal and larvicidal behaviour enables E. vuilleti to develop on C. maculatus instead of D. basalis. E. vuilleti females are able to parasitise the L5 larval stage and the pupa of D. basalis: their larvae therefore feed at the expense of the developing parasitoid. This trophic level is that of hyperparasitism. However, E. vuilleti females rarely practise hyperparasitism on their own L5 larvae and on those of E. orientalis. This behaviour reveals a high behavioural plasticity enabled by intra- and interspecific recognition of parasitoids used as hosts. Hyperparasitism activity in E. orientalis females is higher than that in E. vuilleti females since they hyperparasitise host parasitoids more frequently without preferential species choice. However, E. vuilleti seems to be free from competitive pressure with E. orientalis, as the former penetrates deeply into a grain store contaminated with C. maculatus in contrast to E. orientalis females, which remain on the surface from where they escaped.     

La radiation des échinodermes au Paléozoïque inférieur,l’exemple des blastozoaires

The subphylum Blastozoa is the most abundant and among the most diversified of echinoderm groups during the Early Paleozoic. Reappraisal of their highly diverse anatomies suggests that their superficially incomparable morphologies are actually relatively homogeneous among the major blastozoan clades. Their generic diversity shows two peaks (Drumian, Sandbian), linked by a single origination event during the Cambrian-Lower Ordovician interval. During the lower Middle Ordovician, blastozoans were distributed in distinct provinces, but became progressively more cosmopolitan during the Upper Ordovician. After the Late Ordovician crisis event, blastozoans were restricted to the Laurentian and the Baltic margins.     

Signification des technologies bifaciales au Paléolithique moyen des montagnes de l’Altaï

Previously the single bifacial tools found in different industrial variants of the Altai Middle Paleolithic (Sibiryachikha and Kara-bom) were not considered to be cultural markers that could be used to differentiate the technological/cultural variants, but rather a bright, but situational manifestation of the typological variability, especially in the case of the Sibiryachika assemblages. As a result of recent studies of Chagyrskaya Cave, the key-site of Sibiryachikha, it was found that all the bifaces are made using plano-convex technology. In the Chagyrskaya Cave assemblage all stages of bifacial production have been found: pre-forms, bifacial tools and tools made on the bifacial thinning flakes, accompanied by numerous bifacial thinning flakes and bifacial thinning chips. A preliminary study of the bifaces from the second Sibiryachikha site, Okladnikov Cave evidenced the use of the same plano-convex technology. On the other hand, in the Kara-bom complexes (Kara-Bom, Ust-Karakol-1, Anuy-3), all bifacial tools are made using bi-convex bifacial technology. Thus, the criteria for the technological distinction of bifacial production assume special importance as a cultural marker that allows the differentiation the Altai Middle Paleolithic technological variants. Taking into account the fact that Chagyrskaya Cave and Okladnikov Cave are associated only with Neanderthal remains, it can be assumed that bifacial plano-convexe technology in the Middle Paleolithic of Altai is linked to the appearance and existence of their population in the region.     

Alcaloïdes des feuilles de Holarrhena febrifuga

Leaves of Holarrhena febrifuga from East Africa contain 1·5% of alkaloids, mostly of the amino-3α ceto-20 pregnane-5α type. Six alkaloids have been isolated: funtumine (I), holamine (II), dimethyl- and monomethyl-funtumine (Ia) and (Ib), dimethyl- and monomethylholamine (IIa) and (IIb); these methyl derivatives have been isolated for the first time from natural source. Pregnane-3,20-dione has been isolated from the same leaves.