The uterine immune profile: A method for individualizing the management of women who have failed to implant an embryo after IVF/ICSI.

A unique endometrial immune reaction should occur to promote the human embryo implantation. We postulated that an immune disequilibrium may impact the initial dialogue between the mother and her embryo. In 2012, we set a method of uterine immune profiling for patients with unexplained repeated implantation failures (RIF). The method documents the local Th-1/ Th-2 equilibrium and the recruitment and state of maturation/activation of uNK cells. In function of the disequilibrium observed, personalization of assisted reproductive treatments was suggested. As the concept of personalization in function of the uterine immune profile had never been proposed, a large cohort study and a controlled cohort study were first conducted in RIF patients. 80 % of the RIF patients showed a local disequilibrium if compared to fertile controls. The local disequilibrium was identified in 3 categories: over-immune activation in 45 %, low- local immune activation in 25 % and mixed profile in 10 %. Personalization of treatments in function of the immune profile allowed to restore a live birth rate by 40 % at the following embryo transfer. RIF patients with endometriosis show some particularities regarding their immune profiles. We also suggested that immunotherapy (corticoids, intralipids) may have targeted indications based on a better understanding of the immune type of disequilibrium documented. Personalization of treatments for RIF patients seems to be essential to promote the subsequent live birth rate. The endometrial immune profiling is an innovative method aiming to detect a local immune disequilibrium and, if present, to test preventively its correction under treatment.

Prediction of the fertility of stallion frozen-thawed semen using a combination of computer-assisted motility analysis, microscopical observation and flow cytometry.

Spermatozoa from some stallions do not maintain an acceptable fertility after freezing and thawing. The selection of frozen ejaculates that would be suitable for insemination is mainly based on post-thaw motility, but the prediction of fertility remains limited. A recent study in our laboratory has enabled the determination of a new protocol for the evaluation of fresh stallion semen, combining microscopical observation, computer-assisted motility analysis and flow cytometry, and providing a high level of fertility prediction. The purpose of the present experiment was to perform similar investigations on frozen semen. A panel of tests evaluating a large number of compartments or functions of the spermatozoa was applied to a population of 42 stallions, 33 of which showing widely differing fertilities (17-67% pregnancy rate per cycle [PRC]). Variability was evaluated by calculating the coefficient of variation (CV=SD/mean) and the intra-class correlation or "repeatability" for each variable. For paired variables, mean within-stallion CV% was significantly lower than between-stallion CV%, which was significantly lower than total CV%. Within-ejaculate repeatability, determined by analysing 6 straws for each of 10 ejaculates, ranged from 0.60 to 0.97. Within-stallion repeatability, determined by analysing at least 5 ejaculates for each of 38 stallions, ranged from 0.12 to 0.95. Principal component regression using a combination of 25 variables, including motility, morphology, viability, oxidation level, acrosome integrity, DNA integrity and hypoosmotic resistance, accounted for 94.5% of the variability regarding fertility, and was used to calculate a prediction of the PRC with a mean standard deviation of 2.2. The difference between the observed PRC and the calculated value ranged from -3.4 to 4.2. The 90% confidence interval (90CI) for the prediction of the PRC for the stallions of unknown fertility ranged from 8 to 30 (mean = 17). The best-fit model using only motility variables, evaluated after 10 min at 36 °C and 2 h at 36 °C or room temperature, accounted for only 74.2% of the variability. The difference between the observed PRC and the calculated value ranged from -7.2 to 14. The 90CI for the prediction of the PRC for the stallions of unknown fertility ranged from 23 to 48 (mean = 33). In conclusion, this study demonstrated that an appropriate combination of computer-assisted motility analysis, microscopical observation and flow cytometry can provide a higher prediction of fertility than motility analysis alone.

Development of a new fertility prediction model for stallion semen, including flow cytometry.

Several laboratories routinely use flow cytometry to evaluate stallion semen quality. However, objective and practical tools for the on-field interpretation of data concerning fertilizing potential are scarce. A panel of nine tests, evaluating a large number of compartments or functions of the spermatozoa: motility, morphology, viability, mitochondrial activity, oxidation level, acrosome integrity, DNA integrity, "organization" of the plasma membrane, and hypoosmotic resistance, was applied to a population of 43 stallions, 33 of which showing widely differing fertilities (19%-84% pregnancy rate per cycle [PRC]). Analyses were performed either within 2 hours after semen collection or after 24-hour storage at 4 °C in INRA96 extender, on three to six ejaculates for each stallion. The aim was to provide data on the distribution of values among said population, showing within-stallion and between-stallion variability, and to determine whether appropriate combinations of tests could evaluate the fertilizing potential of each stallion. Within-stallion repeatability, defined as intrastallion correlation (r = between-stallion variance/total variance) ranged between 0.29 and 0.84 for "conventional" variables (viability, morphology, and motility), and between 0.15 and 0.81 for "cytometric" variables. Those data suggested that analyzing six ejaculates would be adequate to characterize a stallion. For most variables, except those related to DNA integrity and some motility variables, results differed significantly between immediately performed analyses and analyses performed after 24 hours at 4 °C. Two "best-fit" combinations of variables were determined. Factorial discriminant analysis using a first combination of seven variables, including the polarization of mitochondria, acrosome integrity, DNA integrity, and hypoosmotic resistance, permitted exact determination of the fertility group for each stallion: fertile, that is, PRC higher than 55%; intermediate, that is, 45% < PRC less than 55%; or subfertile, that is, PRC less than 45%. Linear regression using another combination of 20 variables, including motility, viability, oxidation level, acrosome integrity, DNA integrity, and hypoosmotic resistance, accounted for 94.2% of the variability regarding fertility and was used to calculate a prediction of the PRC with a mean standard deviation of 3.1. The difference between the observed fertility and the calculated value ranged from -4.2 to 5.0. In conclusion, this study enabled to determine a new protocol for the evaluation of stallion semen, combining microscopical observation, computer-assisted motility analysis and flow cytometry, and providing a high level of fertility prediction.

Use of combinations of in vitro quality assessments to predict fertility of bovine semen.

Predicting in vivo fertility of bull ejaculates using in vitro-assessed semen quality criteria remains challenging for the breeding industry. New technologies such as computer-assisted semen analysis (CASA) and flow cytometry may provide accurate and objective methods to improve semen quality control. The aim of this study was to evaluate the relationship between semen quality parameters and field fertility of bull ejaculates. A total of 153 ejaculates from 19 Holstein bulls have been analyzed using CASA (postthawing semen motility and morphology) and several flow cytometric tests, including sperm DNA integrity, viability (estimated by membrane integrity), acrosomal integrity, mitochondria aerobic functionality and oxidation. Samples were analyzed both immediately after thawing and after 4 hours at 37 °C. A fertility value (FV), based on nonreturn rate at 56 days after insemination and adjusted for environment factors, was calculated for each ejaculate. Simple and multiple regressions have been used to correlate FV with CASA and flow cytometric parameters. Significant simple correlations have been observed between some parameters and FV (e.g., straight line velocity [µm/s], r(2) = -0.12; polarized mitochondria sperm (%), r(2) = 0.07), but the relation between simple parameter and FV was too week to predict the fertility. Partial least square procedure identified several mathematical models combining flow cytometer and CASA variables and had better correlations with FV (adjusted r(2) ranging between 0.24 and 0.40 [P < 0.0001], depending on the number of included variables). In conclusion, this study suggests that quality assessment of thawed bull sperm using CASA and flow cytometry may provide a reasonable prediction of bovine semen fertility. Additional work will be required to increase the prediction reliability and promote this technology in routine artificial insemination laboratory practice.

Extreme phenotypic variability of thyroid dysgenesis in six new cases of congenital hypothyroidism due to PAX8 gene loss-of-function mutations.

CONTEXT: Within the last two decades, heterozygous loss-of-function PAX8 mutations have been reported in patients with a wide degree of thyroid gland dysfunction and growth despite the presence of identical mutations. OBJECTIVES: To search for PAX8 mutations in a cohort of patients with congenital hypothyroidism (CH) and various types of thyroid gland defects. DESIGN: A cross-sectional study was conducted in a cohort of patients. SETTING: The French neonatal screening program was used for recruiting patients. PATIENTS: A total of 118 patients with CH, including 45 with familial and 73 with sporadic diseases, were included in this study. The thyroid gland was normal in 23 patients had hypoplasia, 25 had hemithyroid agenesis, 21 had athyreosis, and 21 had ectopy. RESULTS: We found four different PAX8 mutations (p.R31C, p.R31H, p.R108X, and p.I47T) in ten patients (six patients with CH and four family members), two with sporadic and eight with familial diseases. Imaging studies performed in the index cases showed ectopic thyroid gland (n=2), hypoplasia (n=2), eutopic lobar asymmetry (n=1), and eutopic gland compatible with dyshormonogenesis (n=1). The previously reported p.R31C and the novel p.I47T PAX8 mutations are devoid of activity. CONCLUSION: Four different PAX8 mutations were detected in six index patients with CH (ten total subjects). The p.R31C, p.R31H, and p.R108X mutations have been reported. The novel p.I47T PAX8 mutation presented loss of function leading to CH. Thyroid ectopy was observed in two cases of PAX8 (p.R31H) mutation, a finding that has not been reported previously. We observed a high inter-individual and intra-familial variability of the phenotype in PAX8 mutations, underlining that population genetic studies for CH should include patients with various clinical presentations.

GnRH receptor mutations in isolated gonadotropic deficiency.

GnRH and its receptor GnRHR are key regulators of the hypothalamo-pituitary axis. They modulate the secretion of LH and FSH gonadotropins and therefore, the development and maturation of gonads in fetal life as well as after birth. Congenital functional defect of this axis results in isolated hypogonadotropic hypogonadism (IHH). Several natural mutations causing IHH without anosmia have now been identified in GnRHR or GnRH genes. These mutations inactivate GnRHR or its ligand function and cause highly variable phenotypes, ranging from partial to complete gonadotropic deficiencies. The present review describes the published natural GnRHR mutations and tries to correlate them with the corresponding phenotypes according to the different steps of the GnRH system development.

Head, face and neck injuries in hockey: a descriptive analysis.

Patients presenting to the emergency departments in Kingston, Ontario, between 1 October 1992 and 30 April 1993 with head, face, and neck injuries from playing ice hockey, regardless of the age of the player or whether the play was recreational or league, were enrolled in this prospective descriptive case series analysis to document the type and mechanism of injury in relation to the use of protective head and neck gear. A total of 119 such injuries were seen, 84 (71%) of which were lacerations. Players aged 20-34 years were most frequently injured, most commonly through contact with sticks and pucks while wearing helmets but no face shields. Strict enforcement of the rules is required to minimize injuries. Further study is required to determine the reasons for the incomplete protection afforded by helmets and face shields noted here.

[Study and differentiation of some hydrolases active on triglycerides and esters, in anaerobic bacteria, using gas liquid chromatography (author's transl)]. / Etude et différenciation de certaines hydrolases actives sur les triglycérides et les esters, chez les bactéries anaérobies, par la méthode de chromatographie en gaz-liquide.

Anaerobic bacteria are classified using presence or absence of phospholipase and lipase, among other criteria. Techniques are described for the qualitative and quantitative detection of bacterial esterases (carboxylic ester hydrolase) and lipases (triacylglycerol acyl hydrolase) endo- or -exocellular, using gas liquid chromatographic method. Results with representatives anaerobic species are briefly presented.

[Actinomyces odontolyticus, first study in France]. / Actinomyces odontolyticus, première étude réalisée en France.

The authors report the detailed study of a strain of Actinomyces odontolyticus isolated from a pleural liquid. Gas-liquid chromatography can specify the biochemistral metabolism of the germ and set off an endocellular lipidolytic activity.

[Lipase activity of anaerobic bacteria on glycerol-tributyrate determined by gas-liquid chromatography]. / Etude des lipases actives sur le glycérol-tributyrate chez les Bactéries anaérobies par la méthode de chromatographie gaz liquide

Anaerobic bacteria are classified among other criteria by the presence or absence of phospholipase and lipase. The liquid gas chromatographic method detects with great sensibility the lipasic activity of the anaerobic bacteria. A liidolytic action has been demonstrated in Cl. perfringens.

[C-type virus particles in lymphocytes cultured from cows with persistant lymphocytosis. Preliminary results on French cattle stock]. / Présence de particules virales de type C dans des cultures de lymphocytes de bovins présentant une lymphocytose persistante, résultats préliminaires concernant le cheptel bovin francais

C type virus particles were shown in cultures of lymphocytes originating from cows with persistent lymphocytosis. These observations on French cattle are in complete accordance with data previously obtained in the USA. Viral replication is highly stimulated by the addition of phytohemagglutinin in the culture medium. Several morphological and biological properties, specific to the bovine virus differentiate this virus from C type viruses of other species.