The disability associated with hand osteoarthritis is substantial in a cohort of post-menopausal women: the QUALYOR study.

OBJECTIVES: Our primary aims were to assess current prevalence of HOA and the disability associated with this condition, in the group usually most affected, i.e., women older than 55. METHODS: We performed hand radiographs, clinical examination, grip strength measurement, AUSCAN and COCHIN questionnaires in a cohort of postmenopausal women aged at least 55. Radiographic hand OA (RHOA) was defined as at least 2 affected joints among 30, grading 2 or more using the Kellgren Lawrence score but without any HOA symptom. Symptomatic HOA (OA ACR) was defined according to ACR criteria for hand OA. Moderate to severe symptomatic HOA was defined as having OA ACR and AUSCAN total score of >43/100. RESULTS: We enrolled 1,189 participants. The mean age was 71.7 years. Inter-reader reliability of radiographs reading was good (ICC = 0.86) and intra-reader reliability was excellent (ICC = 0.97). Among the 1,189 women, 333 (28.0%) had RHOA, 482 (40.5%) patients fulfilled the ACR criteria for symptomatic HOA and 82 of these (17% of OA ACR population) had moderate to severe symptomatic HOA. The prevalence of symptomatic erosive osteoarthritis was 11.8%. Mean AUSCAN and Cochin scores were higher and grip strength lower in patients with symptomatic HOA compared to patient without HOA. Differences were more noticeable in patients with moderate to severe HOA. CONCLUSIONS: We have assessed disability associated with HOA in greater detail than previously and found that a third of postmenopausal women had RHOA, two fifths had symptomatic HOA and one sixth of symptomatic patients had moderate to severe HOA related disability and a tenth had symptomatic erosive osteoarthritis, representing a substantial burden of disease in our population-based cohort.

Long-term effects in progeny of paternal environment and of gamete/embryo cryopreservation.

In addition to gross malformations, many problems relating to the formation of gametes and embryos can generate, within a continuum of abnormalities, a number of problems that are less evident. On the basis of genetic and/or biochemical or cytological changes, these effects generally appear long after birth as functional difficulties that range from growth changes and altered endocrine functions and cancer to very late behavioural disorders. Such problems may have effects on males and females before conception, on the embryo during gestation, and may also impact on the success of assisted reproduction techniques. For this reason, we have examined the experimental and clinical data that indicate the long-term consequences, for progeny, of iatrogenic and toxic environmental factors on the male reproductive system, and in particular the effect that one specific condition-cryopreservation-may have on gametes and the conceptus. We then focus on the interpretation given to these data which, in general, emphasize the need not only for further experiments to help understand the mechanism of anomalies and increase the level of vigilance in humans, but also to extend follow-up investigations in children.

Post-pubescent to mature fathers: increase in progeny quality?

In the mouse, offspring born from mature fathers exhibit better behavioural performances (for spontaneous activity in both sex and learning capacity in males) than those born from post-pubescent fathers. These behavioural variations are not accompanied by other obvious modifications. They could correspond to what has been observed in man relating to the results in a psychometric test undergone by male progeny born from very young to mature fathers. These data and those which show a decrease in mental performance in rats and human males born from mature to senescent fathers suggest the participation of some genetic factors in these phenomena.

Behavioral teratogenesis: an extension to the teratogenesis of functions.

Major malformations correspond to pathology during the first 2 months of gestation. Thereafter, histological and biochemical abnormalities can result from different negative maternal incidents and, without obvious malformations, change the phenotype of the conceptus. These abnormalities lead to essentially functional disorders often compatible with life and to more or less serious handicaps. All systems, when they arrive at such a stage of development, can be theoretically involved. The central nervous system (CNS) offers a very clear example of such weaknesses because its maturation is particularly long and susceptible to changes even after birth. The steps of this maturation overlap and form a continuum reflected in the pathology. Mental retardation occurring in children born of mothers who were subjected to atomic radiation, or ingested methylmercury from industrial waste, or who suffered the effects of lead, alcohol or tobacco constitutes a clear clinical example. Animal experiments confirm these data, adding pathogenic explanations. These experiments also explore the possible consequences that some medical techniques such as modern reproductive technology, for example embro freezing, can have on the development of the conceptus. Toxic substances or drugs can also be responsible for such abnormalities through a genetic attack on spermatogenesis. Behavioral teratogenesis also opens a larger perspective related to the optimal quality of the conceptus and the determining factors, from stress or dietary factors during apparently normal pregnancy to paternal age at the moment of conception. Finally, given that other systems than the CNS can be involved in histological or biochemical abnormalities, such as the reproductive system, we must ask what other types of functional pathology can be induced by interventions on gametes, the embryo and the fetus. Thus, behavioral teratogenesis leads to the teratogenesis of functions.

Cryopreserved immature mouse oocytes: a chromosomal and spindle study.

PURPOSE: Cryopreservation of human oocytes might provide an alternative approach to freezing supernumerary embryos obtained during IVF. This process, performed on immature denuded prophase 1 mouse oocytes, was investigated. METHODS: We first investigated the capacity of frozen, immature, murine oocytes to continue in vitro maturation after thawing. We then evaluated the risk to offspring from chromosomal damage by cytogenetical and cytological (spindle) analysis. Finally, we attempted to determine the reasons for and the stage of maturation failure. RESULTS: A total of 700 immature oocytes was frozen, 629 (90%) were recovered intact after thawing, and 53% extruded the first polar body, versus 74% for the control group. Freezing was not accompanied by an increase in aneuploidy in maturing oocytes (18 and 15% for thawed and control oocytes, respectively). Consequently, the first meiotic division occurred normally, without an increase in nondisjunction. Spindle analysis demonstrated only a few abnormalities (15 and 2% for thawed and control oocytes, respectively) incompatible with further development. Oocytes arrested during in vitro maturation were mainly at the metaphase I stage (64 and 76% for thawed and control oocytes, respectively). Whereas 17% of thawed oocytes were blocked before the formation of the first meiotic spindle, this never occurred in the control group. CONCLUSIONS: Immature murine oocytes can withstand cryo-preservation, which is encouraging for future human application of this technique.

[Capabilities and limitations of research]. / Possibilités et limites de la recherche.

Assisted human reproduction (AHR) currently solves numerous difficulties caused by sterility in couples. However, it poses some problems whose solutions involve three main directions in research. Fundamental research explores spontaneously occuring phenomena. It shows that over and above common general points of sexual reproduction, mechanisms can differ from one to another. Technical research perfects artificial empirical methods which lead to fertilization, implantation and pregnancy. It shows that success varies from one species to another. Concerning some technologies such as intracytoplasmic sperm injection, successful results began with humans where they seem to be easier to obtain. Research of risks cheks, in animals, the absence of adverse effects of the above techniques for conceptus. It has confirmed the harmfulness of some environmental factors for gametes and zygotes. Moreover, it shows that these negative effects vary between species and within them for strains and sex. For instance, the cryopreservation of mouse embryos leads to changes in the offspring concerning morphophysiological and behavioral features, some of them appearing in elderly subjects. Indeed, these changes vary as a function of strain and sex. These results as a whole show that experimentation on animals can indicate research areas and also give rise to the need for caution. However, if we want to act on human reproduction, they also show, because of the variations in reactivity from one species to another, that the animal model is insufficient and that research in man is essential. From this point of view, a long term follow up of AHR children seems necessary, as well as the need for reflexion concerning the possibility and conditions of research on human embryos.

Chromosome 18 analysis by fluorescence in situ hybridization (FISH) in human blastomeres of abnormal embryos after in vitro fertilization (IVF) attempt.

We performed fluorescence in situ hybridization (FISH) with a chromosome 18-specific probe on human abnormal cleaved embryos, fertilized either by two spermatozoa and exhibiting three pronuclei (3 PN) or normally fertilized and exhibiting two pronuclei (2 PN) with subsequent severe fragmentation and/or blocking. The aim of the study was to evaluate the incidence of chromosome 18 anomalies among these embryos in order to evaluate the FISH efficiency on such material and to obtain more precise and complete data than those obtained with classical cytogenetic analysis. For the 3 PN cleaved embryos, FISH confirmed the frequent regulation towards diploidy (25 per cent) and the high frequency of mosaics (53 per cent). For the 2 PN blocked or damaged embryos, FISH permitted chromosome evaluation, which was otherwise impossible with classical cytogenetic techniques: we also found a high mosaic frequency (45 per cent) with these embryos. If this frequency were the same for normally developing embryos, it would be a major obstacle to the reliability of either chromosomal or genetic preimplantation diagnosis.

Effects of cooling and equilibration in DMSO, and cryopreservation of mouse oocytes, on the rates of in vitro fertilization, development, and chromosomal abnormalities.

In a previous study, we have shown that the cryopreservation of mouse oocytes caused increases in the rates of degeneration and of digynic polyploid embryos, while the fertility of frozen-thawed oocytes was decreased. In this study, we have attempted to determine the different stages in the complete freezing-thawing process which are deleterious for the oocytes and the subsequent zygotes. IVF assays showed that DMSO decreased the fertility of oocytes, whereas cooling to 0 degrees C had no effect. DMSO, used at 0 degrees C, was less deleterious for oocytes. Thus, the prefreezing manipulations seem to be important for the quality and fertility of oocytes. However, neither DMSO nor cooling increased the incidence of chromosomal abnormalities in embryos obtained from inseminated exposed oocytes. Therefore, the increased frequency of polyploidy observed in embryos after the cryopreservation of mouse oocytes must correspond to disruption occurring during the freezing-thawing process.

Long-term effects of embryo freezing in mice.

Embryo cryopreservation does not induce clear-cut anomalies at detectable rates, but several mechanisms exist for nonlethal damage during the freeze-thaw process, and the risk of moderate or delayed consequences has not been extensively investigated. In a long-term study including senescence, we compared cryopreserved and control mice for several quantitative traits. Significant differences were seen in morphophysiological and behavioral features, some of them appearing in elderly subjects. Thus, apart from its immediate toxicity, embryo cryopreservation, without being severely detrimental, may have delayed effects. These results, consistent with other findings, question the neutrality of artificial reproductive technologies and draw attention to the preimplantation stages in developmental toxicology.

Spermatozoa as potential carriers of HIV.

In order to investigate the role of germ cells in the sexual transmission of immunodeficiency virus (HIV), spermatozoa from healthy HIV-seronegative men were incubated in vitro with HIV1. After washing, they were cocultured with peripheral blood leukocytes from seronegative blood donors. Reverse transcriptase assays and p24 antigen tests were performed in culture supernatants. Electron microscopy examination of these HIV-incubated spermatozoa was carried out, as well as the search for CD4 molecules on their surface. Although virus bound to and seemed to enter spermatozoa despite the absence of detectable CD4 epitopes on their surface, no replication of HIV was apparent. However, HIV particles on the surface of spermatozoa were capable of infecting CD4 T lymphocytes. Present results would seem to preclude artificial insemination between an HIV-seropositive man and an HIV-seronegative woman.

Cryopreservation of mouse oocytes: mutagenic effects in the embryo?

We have shown in previous studies that the complete cycle of cryopreservation and prefreezing manipulations increases the degeneration and decreases the fecundability of mouse oocytes. The present study confirms these results. Moreover, we show that the increase of polyploidy previously observed in one-cell zygotes derived from frozen-thawed oocytes persists during the early stages of embryonic development. Furthermore, embryos obtained from frozen oocytes or oocytes exposed to prefreezing manipulations show an increase in the frequency of sister chromatid exchanges. Since the estimation of sister chromatid exchange is a sensitive test of mutagenicity, this suggests that the complete cycle of cryopreservation might alter the oocyte and, more particularly, induce DNA damage.

[Age of the father and development]. / Age du pere et développement.

Testicular ageing affects at the same time the individual and his lineage. In the individual, vascular, endocrine, blood testis barrier and Sertoli cells changes because of age lead a decrease of spermatozoa number and an alteration in their form and motility. These changes lead a gradual decrease of fertility. In the progeny, paternal ageing is responsible for new dominant autosomic mutations which themselves cause different malformations, as achondroplasia, Apert or Recklinghausen disease, Marfan Syndrome etc. and perhaps for certain chromosome X linked recessive mutations as Duchenne myopathy or hemophily A. Moreover, in animal and man, paternal ageing seems responsible for a gradual lowering in the level of progeny cerebral functions. In man, very youthful age is also related to these effects. Thus, the curve corresponding to this phenomenon presents an inverted U-Shape, of which the top corresponds to about thirty years of paternal age. Maternal age does not appear to play a part in this event. On the whole, these results pose the problem of the optimum age for fatherhood.

[The quality of conceptus as a function of father's age]. / La qualité du conceptus en fonction de l'âge du père.

Testicular ageing affects at the same time the individual and his lineage. In the individual, vascular, endocrine, blood testis barrier and Sertoli cells changes because of age lead a decrease of spermatozoa number and an alteration in their form and motility. These changes lead a gradual decrease of fertility. In the progeny, paternal ageing is responsible for new dominant autosomic mutations which themselves cause different malformations and perhaps for certain chromosome X linked recessive mutations. Moreover, in animal and man, paternal ageing seems responsible for a gradual lowering in the level of progeny cerebral functions. In man, very youthful age was also related to these effects. Maternal age did not appear to play a part in this event. On the whole, these results pose the problem of the optimum age for fatherhood.

[Age of the father and development potential]. / Age du pere et aptitude au developpement.

PIP: Testicular aging, like ovarian aging, concerns not just the individual but also the quality of the gametes and hence of the offspring. The 1st signs of testicular aging appear early. Beginning around the age of 30, the vascularization begins to thin, with a progressive decline in the density of the capillaries. The membrane of the seminiferous tubules, an essential element of the hematotesticular barrier, begins to thicken and the number of Sertoli cells begins to decline. Endocrine effects usually appear a decade later, but individual variations are considerable. These modifications are accompanied by a slow decline in the number of sperm and alterations in their morphology and motility. Male fertility declines progressively with age. The quality of the gametes is lower among very young males and increases to a maximum at about age 30. Paternal aging may be responsible for well-defined syndromes in the offspring. Paternal aging has long been recognized as a factor in dominant autosomal mutations causing macroscopic malformations such as achondroplasia, Apert syndrome, Marfan's syndrome, fibrodysplasia ossificans progressiva, and others. The frequency of each disorder is very low, but the total number of recognized disorders of this type exceeds 1000, multiplying the risks so that the .3-.5% risk of anomalies due to paternal aging after 40 is comparable to the risk of trisomy 21 for women aged 35-40. Dominant autosomal mutations can also be responsible for less marked anomalies such as Recklinghausen's neurofibromatosis. Some authors believe that recessive mutations linked to the X chromosome causing hemophilia or Duchenne muscular dystrophy can also result from paternal aging. Some evidence suggests that for a given maternal age, paternal age results in subtle and continuous declines in cerebral functioning. A psychometric study of 1700 military recruits in Nancy, France, in 1985 who were 18 years old showed that sons of very young fathers and of older fathers did less well on the tests. The study is being repeated on 12,000 recruits in the Paris area in 1989-90 to verify the results. Efforts will be made to separate the influence of socioeconomic status and birth order on the results.^ieng

Influence of male sexual rest and oocyte aging on parthenogenesis frequency in mice: cytogenetic analysis after in vitro fertilization.

This study was conducted to evaluate the influence of male sexual rest and oocyte aging on fertilization rate and parthenogenesis frequency after in vitro fertilization of mouse oocytes. We used a comparison between cleavage rates and fertilization rates according to chromosomal analysis of oocytes to estimate the parthenogenesis frequency. Fertilization rate was not impaired by male sexual rest. Parthenogenesis frequency was increased by male sexual rest. This effect was enhanced by a concomitant moderate oocyte aging. It is concluded that cleavage rate could not be considered as a reliable test of fertilization after attempted in vitro fertilization in such conditions.