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.     

La sélection des spermatozoïdes à fort grossissement permet-elle une diminution de la fréquence des aneuploïdies spermatiques?

Severe male infertility concerns two categories of men. Men with abnormal karyotype, who represent 2 to 14% of infertile men and who can produce sperm cells carrying unbalanced chromosomes related to the patients initial chromosomal reorganization inducing a variable risk of transmission of the abnormality to their conceptus. The second category is men with a normal karyotype but an increased rate of spermatic aneuploidy in a context of severe oligo- and/or asthenozoospermia and men from couples in implantation failure. ICSI is the standard Assisted Medical Reproductive technique for most of these 2 categories despite the obvious increased chromosomal risk. This raises the question of how to morphologically identify sperm cells with abnormal chromosome content during ICSI ? Unfortunately, no relationship has yet been found between sperm morphology in the ICSI sperm fraction (×200) and their chromosome content. Nevertheless, since the end of the 1990s, Bartoov’s team has developed MSOME (Motile Sperm Organelle Morphology Examination) consisting of high-power examination of sperm cells up to × 12,250. This technique was indicated for cases of repeated ICSI failures and appeared to increase pregnancy rates. But was this improvement due to better selection of the chromosomal content of sperm cells to be injected? The present study addressed this question by estimating the value of MSOME in the selection of euploid sperm cells in 2 groups of patients known to have an increased rate of sperm aneuploidy. Group 1 was composed of 2 patients with normal karyotype who presented a macrocephalic sperm syndrome with more than 99% of aneuploid sperm. Group 2 was composed of 11 patients with abnormal karyotype: 6 patients with reciprocal translocation and 5 patients with Robertsonian translocation. The purpose of this study was to compare spermatozoa aneuploidy rates in fresh semen, to those obtained after ICSI selection (×200) and MSOME selection (×6000). Three specific steps of the protocol were (1) all sperm cells selected in MSOME were “top sperm cells“ (2) fixation of selected sperm cell (average loss of 15% during FISH washes) (3) FISH results were validated by two different examiners. FISH analysis of X, Y and 18 chromosomes showed that MSOME eliminates polyploid and diploid sperm cells in patients with macrocephalic sperm syndrome, but the 6 sperm cells selected were all haploid and aneuploid. FISH analysis of X, Y and 18 chromosomes of all other patients did not show any influence of the selection method on the aneuploidy rate. For the 5 subjects with a Robertsonian translocation, the global results of FISH analysis paradoxically showed a significant decrease of the euploidy rate in MSOME selection. The global results of FISH analysis for the 6 patients with mutual reciprocal translocations, showed that the various mutual translocations were not modified between whole sperm and the 2 selection methods. On the other hand, a significant decrease of adjacent 1 and 2 segregation frequency was observed between whole sperm and MSOME selection, associated with a significant increase of 3:1 segregation frequency suggesting that the segregations which modify the structure of chromosomes, for example adjacent 1 and 2 segregations, would induce visible morphological modifications selected by MSOME. We hypothesized that the efficacy of spermatic apoptosis could be modulated by morphology but also by the chromosome contents of the sperm cell. In conclusion, MSOME does not provide any guarantee of the normal chromosome contents of the TOP selected sperm cell. However, these results obtained in a small series of patients suggest that MSOME can eliminate some chromosome abnormalities (adj1 and 2) which would alter sperm nuclear structures.     

Sur les ligands stéroïdoïdes de l'androgen binding protein. les énantiomères des Trans-trans etCis-trans perhydrohexestrols et Trans-trans perhydrodicétones

In the framework of a study on nonsteroidal androgens. the authors previously observed that in perhydrohexestrol series, the (±)-[(ci's-4 hydroxycyclohexyl)-4 (trans-4 hydroxycyclohexyl)-hexane] or cis-trans perhydrohexestrol and especially the (±)-(3,4 bis (rans-4-oxocyclohexyl)-hexane) or trans-trans perhyd-rodiketone, there is no affinity for AR (androgen receptor of the prostate) but they bind with high and specific affinity to the testosterone binding site on ABP (epididymal androgen-binding protein). In this work, we describe the preparation, the stereochemistry and biological activities of trans-trans and cis-trans pcrhydrohexestrols and of trans-trans perhydrodiketone as well as their corresponding enantiomers. Biologically the tests AR and ABP arc negative for the trans-trans perhydrohexestrol and of trans-trans perhydrodiketone and for its enantiomers. However for the enantiomers of cis-trans perhydrohexestrol and of trans-trans perhydrodiketone. the affinity of the (+) enantiomer for ABP is superior to that of the racemic and that of the (−) enantiomer. whereas the affinity for AR are zero.Chemically, the stereochemistry of the three racemics has been established especially by X-ray crystallographic analysis or by ¹H n.m.r. The n.m.r. spectra have been analyzed in terras of chemical shifts and coupling constants.     

Utilisation des spermatozoïdes testiculaires en ICSI. Intérêt de la culture in vitro. Revue de la littérature

The number of ICSI cycles performed with testicular spermatozoa has increased dramatically over recent years. However, one of the technical limitations of this approach concerns the extremely reduced motility of testicular spermatozoa. However, increased sperm motility was observed after incubating testicular samples for several hours. Therefore, in order to improve ICSI success rates, several authors have tested the effect of previous in vitro culture. We present a review of the literature on this subject. In vitro culture does not appear to be very useful in cases of obstructive azoospermia, as, apart from possible sperm “maturation” during this culture phase, a high proportion of motile spermatozoa is usually already observed prior to in vitro culture. The benefits of in vitro culture appear to be greater in the case of non-obstructive azoospermia, as when spermatozoa are present on the biopsy, they are usually immobile. However, discordant results have been published: after in vitro culture, spermatozoa have been reported to be either motile or mostly dead. Regardless of the type of azoospermia, the best results are obtained after 3–4 days of in vitro culture. Addition of recombinant FSH to the culture medium also appears to be effective. Cryopreservation of testicular biopsies may also be associated with in vitro culture and the in vitro culture/freezing sequence appears to give better results than the freezing/in vitro culture sequence. Very few studies have reported the results of ICSI using frozen in vitro cultured spermatozoa, as most published studies concern fresh spermatozoa, used after 1–2 days of in vitro culture with satisfactory fertilization and pregnancy rates. In vitro culture of testicular spermatozoa may therefore constitute an interesting research approach to improve the results of ICSI when the number of spermatozoa and/or motility are very low. In addition, in vitro culture of testicular spermatozoa appears to be a good tool to study the mechanisms of acquisition of motility, which are still poorly understood.     

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     

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.     

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.     

Ecophysiologie de quelques Desmidiées I. Un mode d'utilisation des nitrates

Summary The nitrate-reductase activity of some Desmids collected during the stationary phase of culture has been observed in vivo.Within the limits of pH corresponding to the natural biotops, the reduction of nitrates is not followed by any valuable utilisation of the nitrites produced; it is specifically influenced by the pH; it increases with temperature within the experimented limits (from 22° to 38°); increasing also with the concentration of the nitrate up to a saturation value which is higher than the nitrate values observed in nature; the reduction of nitrate is more active in the dark than in the light. Some characteristics of the nitrate-reductase activity seem to be related to some of the ecological aspects of Desmids.     

Une approache pratique des résistances psychologiques aux traitements par injections intracaverneuses (I.I.C.)

Intra-cavernous injection of vasoactive substances has dramatically modified the medical approach of erectile dysfunction. There are psychological and ethical difficulties to this therapeutic procedure, and it is likely that they do contribute to the high drop out rate encountered with this method. According to our clinical experience, we suggest several attitudes in order to facilitate patient's and partner's acceptance:

? There is no opposition between psychotherapy and intra-cavernous injection, they may be regarded as complementary.

? It is advisable to find means to reduce the fear induced by self injection, by reassuring the patient, and making the act easier by choosing the most simple injection material. The pharmacological action must be explained, as well as the side effects.

? Pharmacologically induced erection is often described as artificial, although intra-cavernous injection of vasoactive substances is a symptomatic treatment just as many other symptomatic treatments. It compensates a functional insufficiency, restoring a physiological state very close to normal as far as erection is concerned, but has no effect on sexual excitation, glans intumescence and the development of the ejaculatory climax reflex. The sexuel relation's erotization lays only within the couple, and remains entirely apart from any iatrogenic effect.

? Should the treatment remain secret? Inta-cavernous injection must not systematically be performed withtout the partner knowing about it, although the act should remain discrete and intimate.

One should not encounter psychic or ethical difficulties in erectile dysfunction treatment with intra-cavernous injection of vasoactive substances, provided that it has been properly prescribed, well explained, and that it is part of a psychotherapy. It is doubtless that many refusals or therapy withdrawals are caused by poor or lack of technical initiation, on one hand, and by not taking into account the psychic and ethical aspects of the method, on the other hand.     

Ultrastructure des photorécepteurs de Lineus ruber (O. F. Müller) (Hétéronémertes Lineïdae)

Summary The eyes of the Nemertean worm Lineus ruber are situated on the dorsal face of the animal in two lateral bands in front of the cerebral ganglia. There are 6 or 8 ocelli per animal enclosed in the cephalic parenchyme. They consist of a pigmented eyecup in which 50 photoreceptor cells are present. The photoreceptrice cells have:dendritic processes which are full of mitochondria, neurotubules, vacuoles and only one axial filament;a pericaryon with rough endoplasmic reticulum, a Golgi apparatus and few mitochondria.

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Résumé Les yeux de la Némerte marine Lineus ruber sont situés sur la face dorsale de l'animal, en deux rangées parallèles à l'avant des ganglions cérébroïdes. On en dénombre 6 à 8 localisés dans le parenchyme céphalique. Ils consistent en une cupule pigmentaire entourant une cinquantaine de cellules visuelles qui rejoignent les centres nerveux supérieurs. Pour les cellules photoréceptrices on distingue:un processus dendritique riche en mitochondries, neurotubules, vacuoles et possédant un axe médian;un corps cellulaire où l'on trouve du réticulum endoplasmique rugueux, un appareil de Golgi développé mais peu de mitochondries.