Empreinte parentale et Aide Médicale à la Procréation (AMP)

Medical intervention in procreation is not recent, as the first artificial insemination (AI) was performed more than two centuries ago. However, the interference in the reproductive process with Al is limited. The first major change concerned the possibility of fertilizing oocytesin vitro (IVF) and culture of preimplantation embryos before their transfer to the uterus. In the early nineties, it was shown that direct injection of a single spermatozoon, even abnormal or immature, into an oocyte could result in a viable embryo and child. These techniques expanded very rapidly and 45,000 IVFs, with ICSI in 50% of cases, were performed in France in 2001 (FIVNAT). Although a high incidence of major defects has not been reported, the health status of children born by these techniques is a growing concern. Congenital malformations [Hansenet al., 2002], chromosomal abnormalities [Van Steirteghemet al., 2002], neurological disorders [Stromberget al., 2002] and low birth weight [Schieveet al., 2002] have been observed and discussed, but none of them seems to be statistically much more frequent after assisted reproductive technology (ART). It is important to determine the mechanism of these defects in order to prevent them. These risks may be related to the parents’ health status and to their infertility, but they could also be linked to the techniques used for procreation. Recently, several human and animal studies have suggested an increased risk of imprinting disorders in ART offspring [Debaunet al., 2003; Gicquelet al., 2003; Maheret al., 2003; Hallidayet al., 2004]. Several elements can be considered to be responsible for these defects and each step of reproductive technology could be concerned and must be studied. Priority should be given to confirm the incidence of rare genomic imprinting diseases, such as Beckwith Wiedemann Syndrome and Angelman Syndrome after ART. Should systematic analysis of the methylation status of several imprinted genes therefore be performed to evaluate the respective influence of the use of immature gametes, ovarian stimulation and embryo culture involved in IVF/ICSI? It would also be important to evaluate other epigenetic modifications to determine the role of epigenetic deregulations that could be related to ART.     

Short note Successive approximation in pattern analysis

The analysis of vegetation pattern in semi-arid eucalypt woodland by Harrington et al, (1981), and its recent critique by Dale (1982), place insufficient emphasis on the important role of successive approximation in pattern analysis. A single stage analysis can reveal first order patterns effectively, but is generally ineffective and inefficient in revealing vegetation patterns of higher order, no matter how sophisticated the numerical techniques employed. For a given total analytical effort, a more effective and efficient strategy to reveal such higher order patterns will generally be to partition the overall analytical effort into successive stages of sampling and interpretation, one stage for each order of pattern, with the sampling strategy at each stage based on the patterns determined during previous stages.     

Reproductive biology of the Cape honeybee: a critique of Beekman et al

Laying workers of the Cape honeybee parthenogenetically produce female offspring, whereas queens typically produce males. Beekman et al. confirm this observation, which has repeatedly been reported over the last 100 years including the notion that natural selection should favor asexual reproduction in Apis mellifera capensis. They attempt to support their arguments with an exceptionally surprising finding that A. m. capensis queens can parthenogenetically produce diploid homozygous queen offspring (homozygous diploid individuals develop into diploid males in the honeybee). Beekman et al. suggest that these homozygous queens are not viable because they did not find any homozygous individuals beyond the third larval instar. Even if this were true, such a lethal trait should be quickly eliminated by natural selection. The identification of sex (both with molecular and morphological markers) is possible but notoriously difficult in honeybees at the early larval stages. Ploidy is however a reliable indicator, and we therefore suggest that these "homozygous" larvae found in queen cells are actually drones reared from unfertilized eggs, a phenomenon well known by honeybee queen breeders.     

Response to Midgley: the costs of reproduction cannot differ between the sexes

Many studies have compared the reproductive cost and vegetative growth at a particular time point. In our review (Liu et al. 2021b), we summarized those results but did not compare absolute reproductive costs between the sexes (Hultine et al. 2016; Juvany and Munné-Bosch 2016). Moreover, we did not propose that the observed vegetative and environmental differences between the sexes were the only reasons for differences in sexual functioning, especially in the spreading and receiving of pollen (Midgley 2022). Yet, we need further evidence to support the argument. Previous studies have shown that differences in primary and physiological traits between the sexes strongly depend on the plant species and their environmental conditions, and that they may arise from a number of reasons, such as differences in trait optima of each sex along a series of resource gradients, sexual selection and sex-specific responses to sexual selection (Barrett and Josh 2013; Geber et al. 1999; Juvany and Munné-Bosch 2016; Kohorn et al. 1994; Rabska et al. 2021; Retuerto et al. 2018; Scopece et al. 2021; Wang et al. 2021). In the comment, Midgley (2022) stated that our general argument is that ‘the net reproductive costs are higher for females because they not only flower but must also produce fruits/cones/seeds (Figure 3). Midgley (2022) suggests (Figure 2) that females can ameliorate their higher costs of reproduction by maximizing resource acquisition and resource gain’. However, in our review, we summarized the general opinion and pointed out that this pattern was not universal (see more detail in Liu et al. 2021b). In consistent with previous reviews, our review argues that there is no widespread rule in sex-related differences in the cost of reproduction despite the general opinion that females have higher reproductive costs than males (Darwin 1877; Liu et al. 2021b; Lloyd and Webb 1977). We summarized possible factors causing biased sex ratios in plants, rather than only underpinning the higher net reproductive costs in females than in males (Liu et al. 2021b). Similarly, we proposed possible mechanisms causing sexual differences in responses to biotic stress, rather than underpinning the higher net reproductive costs in females than in males (Liu et al. 2021b), which is also adapted from Núñez-Farfán and Valverde (2020).     

A Program for Monitoring Biological Diversity in the Amazon: An Alternative Perspective to Threat-based Monitoring

Ferraz et al . (2008) indicated the need for planning in biological monitoring in the Amazon, and reviewed what they considered recent progress. The problems and solutions they discuss are well known, and it is unlikely that many biologists would disagree with most of them. However, the authors do not indicate that most of these issues are already being addressed in practice in the Amazon, especially by the Programa de Pesquisas em Biodiversidade (PPBio) of the Brazilian Ministry of Science and Technology. The only major issue about which we do not agree with Ferraz et al . (2008) relates to the design of monitoring programs. The authors recommended tailoring experimental designs to specific threats, which will result in a multitude of idiosyncratic designs and incompatible data sets. We suggest that generic broad-scale designs will allow us to answer more questions with greater confidence, because the investment in each location provides information about regional and global questions. Although specific designs may be more effective in some cases, there are insufficient resources available to install different research infrastructure for every possible threat.     

Retention of cryptic genes in microbial populations

Cryptic genes are silenced genes that can still be reactivated by mutation. Since they can make no positive contribution to the fitness of their carriers, it is not clear why many cryptic genes in microbial populations have not degenerated into useless DNA sequences. Hall et al. (1983) have suggested that cryptic genes have persisted because of occasional strong environmental selection for reactivated genes. The present mathematical study supports their suggestion. It shows that a cryptic gene can be retained without having any selective advantage over a useless DNA sequence, if selection for the reactivated gene occasionally occurs for a substantially long time.      

Current relaxation of selection on the human genome: Tolerance of deleterious mutations on olfactory receptors

Knowledge and understanding about the selective pressures that have shaped present human genetic diversity have dramatically increased in the last few years in parallel with the availability of large genomic datasets. The release of large datasets composed of millions of SNPs across hundreds of genomes by HAPMAP, the Human Genome Diversity Panel, and other projects has led to considerable effort to detect selection signals across the nuclear genome (Coop et al., 2009, Lopez Herraez et al., 2009, Sabeti et al., 2006, Sabeti et al., 2007, Voight et al., 2006). Most of the research has focused on positive selection forces although other selective forces, such as negative selection, may have played a substantive role on the shape of our genome. Here we studied the selective strengths acting presently on the genome by making computational predictions of the pathogenicity of nonsynonymous protein mutations and interpreting the distribution of scores in terms of selection. We could show that the genetic diversity for all the major pathways is still constrained by negative selection in all 11 human populations studied. In a single exception, we observed a relaxation of negative selection acting on olfactory receptors. Since a decreased number of functioning olfactory receptors in human compared with other primates had already been shown, this suggests that the role of olfactory receptors for survival and reproductive success has decreased during human evolution. By showing that negative selection is still relaxed, the present results imply that no plateau of minimal function has yet been reached in modern humans and therefore that olfactory capability might still be decreasing. This is a first clue to present human evolution.     

No phospholipid monolayer-sugar interactions

Studies by a number of workers using the Langmuir film balance have shown that when carbohydrates, such as sucrose or glycerol, are dissolved in a subphase on which a phospholipid is spread, film expansion occurs (Cadenhead & Demchak, 1969; Cadenhead & Bean, 1972; Maggio et al., 1976; Maggio & Lucy, 1978). Recently such effects have been observed again, particularly with the carbohydrates galactose and trehalose (Johnston et al., 1984). The origin of these film expansions was uncertain, and various suggestions have been made to explain them. One idea was that they might be due to interactions which these carbohydrates have with the water molecules close to the polar head groups of the lipids. Recent studies in our two laboratories, described here, show that the magnitude of the expansion effects is variable and that in general they arise from surfactant impurities in the sugars. These impurities are observed in carbohydrates which are reputedly of high grade; the amount of impurity present can vary from batch to batch, and sometimes they can be difficult to remove. Film balance techniques or subphase preparation can mask the detection of minor impurities. The presence of surfactant impurities in reputedly pure carbohydrates needs to be considered in other biochemical and biophysical studies of lipids and cell membranes.     

Hybridization and introgression between "full-fledged species". The case of Parnassius apollo and P. phoebus

Two butterfly species living in the Alps, Parnassius apollo and P. phoebus, frequently hybridize in certain localities of this region. The features of this phenomenon have been previously studied by biometry and starch gel electrophoresis, but some points remained obscure. We present them in a study combining results from cellulose acetate electrophoresis and wing pattern biometry with a determination of the mitochondrial haplotype by a PCR-RFLP analysis in a sample of butterflies from the southern French Alps. It was already known that the male hybrids are fecund and thus that interspecific gene exchange could take place via backcrosses with the parent species. In the present case, combining the identification of mtDNA with the analysis of nuclear genotypes allows us to demonstrate that hybridization can involve both sexes of both species. Moreover, it suggests that at least some female hybrids are not sterile. The impact of Haldane's rule is therefore not very strong in the present case. However, although the prerequisites for introgression between the concerned species are fulfilled, at the level of both nuclear and mitochondrial genomes, no indication of such a phenomenon could be gathered in the studied sample.     

Estimating trees from filtered data: Identifiability of models for morphological phylogenetics

As an alternative to parsimony analyses, stochastic models have been proposed ( [Lewis, 2001] and [Nylander et al., 2004]) for morphological characters, so that maximum likelihood or Bayesian analyses may be used for phylogenetic inference. A key feature of these models is that they account for ascertainment bias, in that only varying, or parsimony-informative characters are observed. However, statistical consistency of such model-based inference requires that the model parameters be identifiable from the joint distribution they entail, and this issue has not been addressed.Here we prove that parameters for several such models, with finite state spaces of arbitrary size, are identifiable, provided the tree has at least eight leaves. If the tree topology is already known, then seven leaves suffice for identifiability of the numerical parameters. The method of proof involves first inferring a full distribution of both parsimony-informative and non-informative pattern joint probabilities from the parsimony-informative ones, using phylogenetic invariants. The failure of identifiability of the tree parameter for four-taxon trees is also investigated.     

Evolutionarily stable strategies and short-term selection in Mendelian populations re-visited

This note concerns a one locus, two allele, random mating diploid population, subject to frequency-dependent viability selection. It is already known that in such a population, any evolutionarily stable strategies (ESS), if only accessible by the genotype-to-phenotype mapping, is the phenotypic image of a stable genetic equilibrium (Eshel, I. 1982. Evolutionarily stable strategies and viability selection in Mendelian populations. Theor. Popul. Biol. 22(2), 204-217; Cressman et al. 1996. Evolutionary stability in strategic models of single-locus frequency-dependent viability selection. J. Math. Biol. 34, 707-733). The opposite is not true. We find necessary and sufficient parametric conditions for global convergence to the ESS, but we also demonstrate conditions under which, although a unique, genetically accessible ESS exists, there is another, "non-phenotypic" genetically stable equilibrium.     

Mice lacking a CDK inhibitor, p57Kip2, exhibit skeletal abnormalities and growth retardation

p57Kip2, one of the cyclin-dependent kinase (CDK) inhibitors, has been suggested to be a tumor suppressor candidate. To elucidate its biological roles in mouse development and tumorigenesis, we created p57Kip2-deficient mice. The p57Kip2-deficient mice exhibited a cleft palate and defective bone formation resulting in severe dyspnea. Most of the p57Kip2-deficient mice died within 24 h after birth, while about 10% of them survived beyond the weaning period. All of the surviving mice showed severe growth retardation. The males showed immaturity of the testes, prostate and seminal vesicles, and the females showed vaginal atresia, immaturity of the uterus, and an increased number of atretic follicles. Although Yan et al. and Zhang et al. have already reported p57Kip2-deficient mice, they could not investigate the phenotypes of the surviving p57Kip2-deficient mice. Also, most of the symptoms of Beckwith-Wiedemann syndrome could not be reproduced in the mutant mice. Embryonic fibroblasts prepared from p57Kip2-deficient mice showed no differences in the proliferation rate and saturation density, suggesting that G1 arrest is largely independent of p57Kip2 function. Our results suggest that p57Kip2 plays a critical role in development, but do not support the hypothesis that the p57Kip2 gene is a tumor-suppressor gene or is responsible for Beckwith-Wiedemann syndrome.     

The Effects of Lethal Selection on the EST-6 to PGM Region of Chromosome 3 in DROSOPHILA MELANOGASTER

A powerful means of studying the effects of selection on chromosome segments in Drosophila melanogaster has been described by Clegg et. al. (1976, 1978). This method utilizes a recessive lethal, dominant visible allele whose selection dynamics can be accurately modelled to predict the fates of nonlethal alleles at linked loci. Results of these experiments indicate that strong epistatic interactions among loci occur that effect fitnesses associated with gametic types in the basal region of chromosome 3. We have used similar methods in studying a different segment of chromosome 3, that spanned by Est-6 (3-36.8) and Pgm (3-43.4), with the aim of determining whether the results of Clegg and his colleagues could be reproduced when a different region was studied. Our experiment showed that selection did operate on a region of the chromosome marked by Pgm, but that no evidence of selection at loci marked by Est-6 was apparent. Weak evidence for epistatic interactions among loci within the marked region was also found. Three possible explanations for the discrepancies between our experiments and those of Clegg et al. are suggested. First, the geographically homogeneous origin of our populations may preclude selectively significant changes as a result of recombination. Second, the results seen by Clegg et al. may have been unique to the regions they studied, which included the basal heterochromatin of the chromosomes. Finally, the three loci employed may not adequately mark the unit of selection, so that actual departures from predictions of selective neutrality may not have been apparent.     

p24 proteins, intracellular trafficking, and behavior: Drosophila melanogaster provides insights and opportunities

The p24 transmembrane proteins, also known as EMP24/GP25 (endomembrane protein precursor of 24kD (Schimmoller et al., 1995)) proteins, are components of coat protein (COP)-coated vesicles and are present in species as diverse as fungi, plants, flies, worms, and mammals, indicating that they have important conserved functions. Genetic, molecular, and biochemical characterization of these proteins and the loci that encode them has provided insights into their potential cellular roles, including postulated functions in vesicle cargo protein selection and sorting, COPI and COPII vesicle formation and budding, and quality control of proteins that mature through the secretory pathway. Recently, the first mutations in a Drosophila melanogaster p24 gene have been isolated and characterized. These alleles produce an interesting behavioral phenotype in females, affecting their ability to oviposit. This identification and mutant characterization of a p24 locus in Drosophila will pave the way for a better understanding of cell-type-specific functions and interactions among p24 proteins.     

Plant transformation by microinjection techniques

Several techniques have been developed for introducing cloned genes into plant cells. Vectorless delivery systems such as PEG-mediated direct DNA uptake (e.g. Pasz-kowski et al. 1984), electroporation (e.g. Shillito et al. 1985), and fusion of protoplasts with liposomes (Deshayes et al. 1985) are routinely used in many experiments (see several chapters of this issue). A wide range of plant species, dicotyledonous as well as monocotyledonous, has been transformed by these vectorless DNA transfer systems. However, the availability of an efficient protoplast regeneration system is a prerequisite for the application of these techniques. For cells with intact cell walls and tissue explants the biological delivery system of virulent Agrobacterium species has been routinely used (for review see Fraley et al. 1986). However, the host range of Agrobacterium restricts the plant species, which can be transformed using this vector system. In addition, all these methods depend on selection systems for recovery of transformants. Therefore a selection system has to be established first for plant species to be transformed. The microinjection technique is a direct physical approach, and therefore host-range independent, for introducing substances under microscopical control into defined cells without damaging them. These two facts differentiate this technique from other physical approaches, such as biolistic transformation and macroinjection (see chapters in this issue). In these other techniques, damaging of cells and random manipulation of cells without optical control cannot be avoided so far. In recent years microinjection technology found its application in plant sciences, whereas this technique has earlier been well established for transformation of animal tissue culture cells (Capecchi 1980) and the production of transgenic animals (Brin-ster et al. 1981, Rusconi and Schaffner 1981). Furthermore, different parameters affecting the DNA transfer via microinjection, such as the nature of microinjected DNA, and cell cycle stage, etc, have been investigated extensively in animal cells (Folger et al. 1982, Wong and Capecchi 1985), while analogous experiments on plant cells are still lacking.     

THE LIFE CYCLE AND TOXICITY OF PFIESTERIA PISCICIDA REVISITED1

Despite use of excellent molecular techniques, Litaker et al. (2002) cannot provide insights about the life history of toxic Pfiesteria piscicida because they showed no data in support of having used toxic strains; rather they presented evidence that they used non‐inducible strains. Litaker et al. did not find amoeboid stages or a chrysophyte‐like cyst stage in several cultures and unequivocally concluded that the stages do not exist in all P. piscicida strains. Thus, they did not consider the tenet that absence of evidence does not constitute proof of absence. Apparent discrepancies between the research by Litaker et al. and previous research on Pfiesteria can be resolved as follows: First, Litaker et al. did not use toxic strains. We have reported findings (similar to Litaker et al.) showing few amoeboid transformations in non‐inducible strains, which manifest some but not all of the forms that have been documented in some toxic strains. We, and others, have documented active toxicity to fish, transformations to amoebae, and chrysophyte‐like cysts in some clonal toxic strains. Second, the data from several recent publications, which were available but not mentioned by Litaker et al. or by Coats (2002) in accompanying commentary, have verified P. piscicida amoebae, chrysophyte‐like cysts, and other stages in some toxic strains through a combination of approaches including PCR data from clonal cultures.     

Water-flea males from the netherworld

Simple treatments with hormones could unlock the expression of complex phenotypes not known to occur in nature. Using this method, Kim et al. recently obtained males from all-female populations of water fleas. The novel characters revealed by this work can be used in taxonomic identification and phylogenetic inference. Additionally, these "resurrected" males offer insights into the conservation of traits that are not exposed to natural selection.     

Is a "color effect" demonstrated for hair analysis of carbamazepine?

Psillakis et al. have reported on the concentration of carbamazepine recovered from hair samples collected from patients receiving this anti-seizure medication under medical supervision and determined that their was a high correlation between dose and quantitation of recovered analyte. The analyte was identified by two techniques, FPIA (Abbott TDx) and HPLC and the correlation was high for both procedures. In the literature on hair analysis some have suggested that analyte concentration in hair is critically dependent on hair color. In reporting their data Psillakis et al. reported the hair color of each patient but made no attempt to analyze their results in relation to color. This article performs a secondary analysis of the Psillakis et al. data in order to determine whether there is a hair color effect discernible in the recovery of carbamazepine from hair. Analysis of this data set for both the FPIA and HPLC by one-way analysis of variance fails to identify a color effect at p = .05. Weighting the data for per-patient dosage values fails to discern a color effect. Examination of all possible two-color comparisons also fails to identify a statistically significant effect for any subset of combinations. These data suggest that carbamazepine does not exhibit a color effect when using either FPIA or HPLC assay methods.     

Biomarkers in Spinal Cord Injury: from Prognosis to Treatment

Spinal cord injury (SCI) is considered an incurable condition, having a heterogenous recovery and uncertain prognosis. Therefore, a reliable prediction of the improvement in the acute phase could benefit patients. Physicians are unanimous in insisting that at the initial damage of the spinal cord (SC), the patient should be carefully evaluated in order to help selecting an appropriate neuroprotective treatment. However, currently, neurologic impairment after SCI is measured and classified by functional examination. The identification of prognostic biomarkers of SCI would help to designate SC injured patients and correlate to diagnosis and correct treatment. Some proteins have already been identified as good potential biomarkers of central nervous system injury, both in cerebrospinal fluid (CSF) and blood serum. However, the problem for using them as biomarkers is the way they should be collected, as acquiring CSF through a lumbar puncture is significantly invasive. Remarkably, microRNAs (miRNAs) have emerged as interesting biomarker candidates because of their stability in biological fluids and their tissue specificity. Several miRNAs have been identified to have their expressions altered in SCI in many animal models, making them promising candidates as biomarkers after SCI. Moreover, there are yet no effective therapies for SCI. It is already known that altered lysophospholipids (LPs) signaling are involved in the biology of disorders, such as inflammation. Reports have demonstrated that LPs when locally distributed can regulate SCI repair and key secondary injury processes such as apoptosis and inflammation, and so could become in the future new therapeutic approaches for treating SCI.     

Study of the distribution pattern of Schistosoma haematobium egg antigens recognised by six different monoclonal antibodies in the parasite and the host

Recently a new panel of monoclonal antibodies was developed against soluble egg antigens in the hatching fluid of Schistosoma mansoni. These antibodies have been used to develop an improved ELISA for the detection of circulating soluble egg antigens in serum and urine that would have a higher sensitivity in the immunodiagnosis of S. mansoni infections. Although these antibodies showed no improvement in the immunodiagnosis of S. mansoni infections compared with egg antigen-based ELISAs already described (Nourel Din et al., 1994a), they may have a potential role in the identification of S. haematobium infections. This study has looked into the immunolocalisation of S. haematobium egg antigens in both the parasite and the host as recognised by four newly developed monoclonal antibodies (290-2D9-A, 290-2E6-A, 290-2H12-A and 290-4A8-A) and two already described antibodies (114-5B1-A and 114-4D12-A). The antibodies 114-5B1-A and 114-4D12-A appeared to have in S. haematobium eggs a similar staining pattern when compared to S. mansoni eggs. The antibodies prepared against the hatching fluid showed a characteristic signal, especially 290-2E6-A. These antibodies recognised a component originating from the lateral glands of the miracidium. In the host a similar immunohistochemical tissue localisation pattern (mainly phagocytising reticulo-endothelial cells) was seen as previously described for S. mansoni infected hamsters.