Successful pregnancy rates amongst patients undergoing ovarian tissue cryopreservation for non-malignant indications: A systematic review and meta-analysis.

BACKGROUND: Ovarian tissue cryopreservation (OTC) is a fertility preservation method that has been clinically applied for almost 30 years. Studies specifically evaluating patients presenting with non-malignant indications for OTC and their subsequent pregnancy rates are limited. OBJECTIVE: To summarise the evidence on the rates of successful pregnancy amongst women who have undergone OTC for non-malignant indications. METHODS: A systematic review with meta-analysis (PROSPERO registration CRD42022307925) was conducted to investigate the pregnancy outcomes of patients who have undergone ovarian tissue cryopreservation for non-malignant indications. Articles published in EMBASE and Ovid MEDLINE before October 2022 were screened for inclusion based on the following criteria: original human studies pertaining to OTC with a defined non-malignant cohort and pregnancy outcomes. The successful pregnancy rates were pooled with a random-effects model of double-arcsine transformed proportions. Sensitivity analysis involved pooling the results of studies with a low risk of bias after being assessed with NIH tools. RESULTS: The database search retrieved 3,225 results, of which 16 were included in the meta-analysis. The pooled successful pregnancy rate was 23.52 % (16 studies, 95 % CI 6.48 to 44.79 %). When subgroup analysis of study types was performed, the successful pregnancy rate was higher amongst case series (47.02 %, 9 studies, 95 % CI 6.98 to 89.00 %) than cohort studies (14.64 %, 7 studies, 95 % CI 3.59 to 29.78 %). Sensitivity analysis limited to studies at low risk of bias revealed a similar pooled successful pregnancy rate of 23.35 % (12 studies, 95 % CI 2.50 to 51.96 %). CONCLUSIONS: Approximately one quarter of women who underwent OTC for non-malignant indications had a successful pregnancy. These findings are clinically important for fertility preservation counselling by providing greater evidence for more informed care.

Embryo selection through artificial intelligence versus embryologists: a systematic review.

STUDY QUESTION: What is the present performance of artificial intelligence (AI) decision support during embryo selection compared to the standard embryo selection by embryologists? SUMMARY ANSWER: AI consistently outperformed the clinical teams in all the studies focused on embryo morphology and clinical outcome prediction during embryo selection assessment. WHAT IS KNOWN ALREADY: The ART success rate is ∼30%, with a worrying trend of increasing female age correlating with considerably worse results. As such, there have been ongoing efforts to address this low success rate through the development of new technologies. With the advent of AI, there is potential for machine learning to be applied in such a manner that areas limited by human subjectivity, such as embryo selection, can be enhanced through increased objectivity. Given the potential of AI to improve IVF success rates, it remains crucial to review the performance between AI and embryologists during embryo selection. STUDY DESIGN SIZE DURATION: The search was done across PubMed, EMBASE, Ovid Medline, and IEEE Xplore from 1 June 2005 up to and including 7 January 2022. Included articles were also restricted to those written in English. Search terms utilized across all databases for the study were: ('Artificial intelligence' OR 'Machine Learning' OR 'Deep learning' OR 'Neural network') AND ('IVF' OR 'in vitro fertili*' OR 'assisted reproductive techn*' OR 'embryo'), where the character '*' refers the search engine to include any auto completion of the search term. PARTICIPANTS/MATERIALS SETTING METHODS: A literature search was conducted for literature relating to AI applications to IVF. Primary outcomes of interest were accuracy, sensitivity, and specificity of the embryo morphology grade assessments and the likelihood of clinical outcomes, such as clinical pregnancy after IVF treatments. Risk of bias was assessed using the Modified Down and Black Checklist. MAIN RESULTS AND THE ROLE OF CHANCE: Twenty articles were included in this review. There was no specific embryo assessment day across the studies-Day 1 until Day 5/6 of embryo development was investigated. The types of input for training AI algorithms were images and time-lapse (10/20), clinical information (6/20), and both images and clinical information (4/20). Each AI model demonstrated promise when compared to an embryologist's visual assessment. On average, the models predicted the likelihood of successful clinical pregnancy with greater accuracy than clinical embryologists, signifying greater reliability when compared to human prediction. The AI models performed at a median accuracy of 75.5% (range 59-94%) on predicting embryo morphology grade. The correct prediction (Ground Truth) was defined through the use of embryo images according to post embryologists' assessment following local respective guidelines. Using blind test datasets, the embryologists' accuracy prediction was 65.4% (range 47-75%) with the same ground truth provided by the original local respective assessment. Similarly, AI models had a median accuracy of 77.8% (range 68-90%) in predicting clinical pregnancy through the use of patient clinical treatment information compared to 64% (range 58-76%) when performed by embryologists. When both images/time-lapse and clinical information inputs were combined, the median accuracy by the AI models was higher at 81.5% (range 67-98%), while clinical embryologists had a median accuracy of 51% (range 43-59%). LIMITATIONS REASONS FOR CAUTION: The findings of this review are based on studies that have not been prospectively evaluated in a clinical setting. Additionally, a fair comparison of all the studies were deemed unfeasible owing to the heterogeneity of the studies, development of the AI models, database employed and the study design and quality. WIDER IMPLICATIONS OF THE FINDINGS: AI provides considerable promise to the IVF field and embryo selection. However, there needs to be a shift in developers' perception of the clinical outcome from successful implantation towards ongoing pregnancy or live birth. Additionally, existing models focus on locally generated databases and many lack external validation. STUDY FUNDING/COMPETING INTERESTS: This study was funded by Monash Data Future Institute. All authors have no conflicts of interest to declare. REGISTRATION NUMBER: CRD42021256333.

DNA repair and response to sperm DNA damage in oocytes and embryos, and the potential consequences in ART: a systematic review.

Sperm DNA damage is considered a predictive factor for the clinical outcomes of patients undergoing ART. Laboratory evidence suggests that zygotes and developing embryos have adopted specific response and repair mechanisms to repair DNA damage of paternal origin. We have conducted a systematic review in accordance with guidelines from Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) to identify and review the maternal mechanisms used to respond and repair sperm DNA damage during early embryonic development, how these mechanisms operate and their potential clinical implications. The literature search was conducted in Ovid MEDLINE and Embase databases until May 2021. Out of 6297 articles initially identified, 36 studies were found to be relevant through cross referencing and were fully extracted. The collective evidence in human and animal models indicate that the early embryo has the capacity to repair DNA damage within sperm by activating maternally driven mechanisms throughout embryonic development. However, this capacity is limited and likely declines with age. The link between age and decreased DNA repair capacity could explain decreased oocyte quality in older women, poor reproductive outcomes in idiopathic cases and patients who present high sperm DNA damage. Ultimately, further understanding mechanisms underlying the maternal repair of sperm DNA damage could lead to the development of targeted therapies to decrease sperm DNA damage, improved oocyte quality to combat incoming DNA insults or lead to development of methodologies to identify individual spermatozoa without DNA damage.

Female ageing affects the DNA repair capacity of oocytes in IVF using a controlled model of sperm DNA damage in mice.

STUDY QUESTION: Does female ageing have a negative effect on the DNA repair capacity of oocytes fertilised by spermatozoa with controlled levels of DNA damage? SUMMARY ANSWER: Compared to oocytes from younger females, oocytes from older females have a reduced capacity to repair damaged DNA introduced by spermatozoa. WHAT IS KNOWN ALREADY: The reproductive lifespan in women declines with age predominantly due to poor oocyte quality. This leads to decreased reproductive outcomes for older women undergoing assisted reproductive technology (ART) treatments, compared to young women. Ageing and oocyte quality have been clearly associated with aneuploidy, but the range of factors that influence this change in oocyte quality with age remains unclear. The DNA repair activity prior to embryonic genomic activation is considered to be of maternal origin, with maternal transcripts and proteins controlling DNA integrity. With increasing maternal age, the number of mRNAs stored in oocytes decreases. This could result in diminished efficiency of DNA repair and/or negative effects on embryo development, especially in the presence of DNA damage. STUDY DESIGN, SIZE, DURATION: Oocytes from two age groups of 30 super-ovulated female mice (young: 5-8 weeks old, n = 15; old: 42-45 weeks old, n = 15) were inseminated with sperm from five males with three different controlled DNA damage levels; control: ≤10%, 1 Gray (Gy): 11-30%, and 30 Gy: >30%. Inseminated oocytes (young: 125, old: 78) were assessed for the formation of zygotes (per oocyte) and blastocysts (per zygote). Five replicates of five germinal vesicles (GVs) and five MII oocytes from each age group were analysed for gene expression. The DNA damage response (DDR) was assessed in a minimum of three IVF replicates in control and 1 Gy zygotes and two-cell embryos using γH2AX labelling. PARTICIPANTS/MATERIALS, SETTING, METHODS: Swim-up sperm samples from the cauda epididymidis of C57BL6 mice were divided into control (no irradiation) and 1- and 30-Gy groups. Treated spermatozoa were irradiated at 1 and 30 Gy, respectively, using a linear accelerator Varian 21iX. Following irradiation, samples were used for DNA damage assessment (Halomax) and for insemination. Presumed zygotes were cultured in a time-lapse incubator (MIRI, ESCO). Gene expression of 91 DNA repair genes was assessed using the Fluidigm Biomark HD system. The DNA damage response in zygotes (6-8 h post-fertilisation) and two-cell embryos (22-24 h post-fertilisation) was assessed by immunocytochemical analysis of γH2AX using confocal microscopy (Olympus FV1200) and 3D volumetric analysis using IMARIS software. MAIN RESULTS AND THE ROLE OF CHANCE: The average sperm DNA damage for the three groups was statistically different (control: 6.1%, 1 Gy: 16.1%, 30 Gy: 53.1%, P < 0.0001), but there were no significant differences in fertilisation rates after IVF within or between the two age groups [(young; control: 86.79%, 1 Gy: 82.75%, 30 Gy: 76.74%) (old; control: 93.1%, 1 Gy: 70.37%, 30 Gy: 68.18%) Fisher's exact]. However, blastocyst rates were significantly different (P < 0.0001) among the groups [(young; control: 86.95%, 1 Gy: 33.33%, 30 Gy: 0.0%) (old; control: 70.37%, 1 Gy: 0.0%, 30 Gy: 0.0%)]. Between the age groups, 1-Gy samples showed a significant decrease in the blastocyst rate in old females compared to young females (P = 0.0166). Gene expression analysis revealed a decrease in relative expression of 21 DNA repair genes in old GV oocytes compared to young GV oocytes (P < 0.05), and similarly, old MII oocytes showed 23 genes with reduced expression compared to young MII oocytes (P < 0.05). The number of genes with decreased expression in older GV and MII oocytes significantly affected pathways such as double strand break (GV: 5; MII: 6), nucleotide excision repair (GV: 8; MII: 5) and DNA damage response (GV: 4; MII: 8). There was a decreased DDR in zygotes and in two-cell embryos from old females compared to young regardless of sperm treatment (P < 0.05). The decrease in DNA repair gene expression of oocytes and decreased DDR in embryos derived from older females suggests that ageing results in a diminished DNA repair capacity. LARGE-SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: Ionising radiation was used only for experimental purposes, aiming at controlled levels of sperm DNA damage; however, it can also damage spermatozoa proteins. The female age groups selected in mice were intended to model effects in young and old women, but clinical studies are required to demonstrate a similar effect. WIDER IMPLICATIONS OF THE FINDINGS: Fertilisation can occur with sperm populations with medium and high DNA damage, but subsequent embryo growth is affected to a greater extent with aging females, supporting the theory that oocyte DNA repair capacity decreases with age. Assessment of the oocyte DNA repair capacity may be a useful diagnostic tool for infertile couples. STUDY FUNDING/COMPETING INTEREST(S): Funded by the Education Program in Reproduction and Development, Department of Obstetrics and Gynaecology, Monash University. None of the authors has any conflict of interest to report.

Male ageing is negatively associated with the chance of live birth in IVF/ICSI cycles for idiopathic infertility.

STUDY QUESTION: Is male age associated with the clinical outcomes of IVF/ICSI cycles for idiopathic infertility after adjustment for female age? SUMMARY ANSWER: Male ageing is negatively associated with clinical IVF/ICSI outcomes in couples with idiopathic infertility independent of female age. WHAT IS KNOWN ALREADY: The effect of male age on the outcomes of infertility treatments is controversial and poorly explored. In contrast, fertility is known to decline significantly with female age beyond the mid-30s, and reduced oocyte quality plays an important role. The negative effect of male age on sperm quality is largely associated with an increasing susceptibility to sperm DNA damage. Although increasing maternal age has been linked with poorer oocyte quality, studies on the effect of male age have disregarded the need to control for female age making it difficult to define clearly the role of male age in infertile couples. STUDY DESIGN, SIZE, DURATION: This retrospective cohort study analysed 2425 cycles of couples with idiopathic infertility selected from a total of 24 411 IVF/ICSI cycles performed at Monash IVF in Australia between 1992 and 2017. The primary outcome was live birth and secondary outcomes were clinical pregnancy and miscarriage. PARTICIPANTS/MATERIALS, SETTING, METHODS: Couples with primary/secondary infertility who underwent IVF/ICSI cycles with male partners classified as normozoospermic were selected (inclusion criteria). Couples in which the female partner had endometriosis, tubal factors, polycystic ovarian syndrome, ovarian hyperstimulation syndrome, poor responders (≤3 mature oocytes retrieved) and couples with more than 15 cumulus oocyte complexes retrieved or who used cryopreserved gametes were excluded. Binary logistic multilevel modelling was used to identify the effect of male age and female age on clinical outcomes after controlling for confounding factors. Male age and female age were examined as continuous and categorical (male age: <40, 40-44, 45-49, 50-54, ≥55; female age:<30, 30-34, 35-39, ≥40) predictors. MAIN RESULTS AND THE ROLE OF CHANCE: There was a negative effect of male age and female age on live birth as odds ratios (OR) with 95% CI for each additional year of age (OR-male age: 0.96 [0.94-0.98]; OR-female age: 0.90 [0.88-0.93] P < 0.001). Potential interactions with male age such as type of treatment (IVF/ICSI), embryo transfer day (Day 3/Day 5) and female age did not have significant associations with outcomes (P > 0.05). Secondary outcomes showed a significant reduction in the odds of clinical pregnancy (OR-male age: 0.97 [0.96-0.99]; OR-female age: 0.92 [0.89-0.94] P < 0.001) and an increase in the odds of miscarriage with older age: male age (OR: 1.05 [1.01-1.08]; P = 0.002); female age (OR: 1.11 [1.05-1.18]; P < 0.001). Worse outcomes were associated with more cycles (clinical pregnancy-OR: 0.96 [0.93-0.99] P = 0.03; live birth-OR: 0.96 [0.92-0.99] P = 0.023) while more inseminated oocytes were associated with better outcomes (clinical pregnancy-OR: 1.06 [1.03-1.06] P < 0.001; live birth-OR: 1.07 [1.04-1.11] P < 0.001). Analyses for age categories showed a gradual worsening of clinical outcomes with increasing male age, with a significantly worse live birth and clinical pregnancy outcomes in males aged older than 50 years compared to males younger than 40 years (P < 0.05). LIMITATIONS, REASONS FOR CAUTION: This study is limited to the information on confounding factors included. The study may also be limited in its generalizability to a wider population due the strict selection criteria. Age as a category could potentially result in residual confounding due to categorizing a continuous variable. WIDER IMPLICATIONS OF THE FINDINGS: This study provides information for counselling of couples with idiopathic infertility. STUDY FUNDING/COMPETING INTEREST(S): Funded by the Education Program in Reproduction and Development, Department of Obstetrics and Gynaecology, Monash University. None of the authors has any conflict of interest to report. TRIAL REGISTRATION NUMBER: N/A.

Gonadotropin-induced puberty does not impair reproductive performance of gilts over three parities.

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty-nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post-weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning-to-oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.

Differences in maternal plane of nutrition and body condition during late gestation coupled with estrus synchronization at weaning do not result in differences in embryonic development at 4 days of gestation.

This study was conducted to assess the effects of dietary energy in late pregnancy and hormone therapy at weaning on plasma metabolite profile, litter performance, reproductive parameters, and embryo viability in the second pregnancy. A total of 23 first-parity sows at 75 d of pregnancy were randomly allocated to 4 treatments. Treatments were factorial (2 × 2) combinations of 2 nutritional strategies [standard-energy feed (SEF) and high-energy feed (HEF)] and 2 hormone therapies [600 IU eCG and 2.5 mg swine LH 72 h later (HO) and no hormone (WH)]. Sows were weighed weekly from 75 d of pregnancy until 3 d before farrowing; 1 d after farrowing; 7, 14, and 21 d into lactation; and at weaning. Back fat (BF) was measured at 75 d of pregnancy, 3 d before farrowing, and at weaning. Average daily gain and ADFI were also calculated. Plasma metabolites were analyzed after 82, 89, 96, and 103 d of pregnancy, at farrowing, and after 7, 14, and 21 d of lactation. Embryo viability was assessed after 4.55 d of second pregnancy. During pregnancy, HEF-treated sows displayed greater BW (P < 0.05) compared with SEF-treated females, but no differences were observed during lactation. There were no differences in BW of the piglets caused by the treatments. High-energy-treated females showed superior BF (P > 0.05) in all periods; however, significant differences were detected only at the prefarrowing measurement (P < 0.05). No differences in ADFI were observed during lactation. The SEF group showed positive ADG, whereas the HEF group showed negative ADG (0.216 vs. -0.266 kg/d for SEF and HEF, respectively; P < 0.05). High-energy-treated sows presented greater concentrations of total cholesterol after 89 and 103 d of pregnancy and greater concentrations of high-density lipid cholesterol (HDL) after 89 and 96 d. At farrowing and 14 and 21 d of lactation, NEFA concentrations were greater (P < 0.05) in the HEF group. After hormone treatment, no differences were observed on weaning-to-estrus intervals and estrus duration. Greater mobilization of body reserves observed in the HEF group during lactation did not affect reproductive performance negatively, suggesting that metabolic status was adequate for the first lactational catabolism.

[Serological and epidemiological features of canine American tegumentary Leishmaniasis from Maricá, Rio de Janeiro, Brazil]. / Aspectos sorológicos e epidemiológicos da leishmaniose tegumentar americana canina em Maricá, Rio de Janeiro, Brasil.

A sample of 261 dogs from 134 households located in a periurban area of Maricá, Rio de Janeiro, Brazil, was studied to evaluate serologic reactions and active infection of american tegumentary leishmaniasis (ATL). Eight dogs presented lesions suggestive of ATL, but this was isolated in only three. Using ELISA, 24.5% (64/261) of the dogs studied were positive (sensitivity = 66% and specificity = 76%), associated with isolation in 2 animals and 0.4% (1/261) by indirect immunofluorescence assay (IIF) with no association with isolation. In order to reduce the unspecific reactions to ELISA, a second criterion was used to obtain the cutoff (sensitivity = 33% and specificity = 93%), resulting in a reactivity of 6.9 % (18/261) associated to isolation in a single animal. As observed in this study, serologic results by IIF were not associated with active infection and ELISA showed high unspecific reactions, indicating that the serologic reactions alone are not recommended for ATL diagnosis. ATL scars were been observed in 7 persons in the region and active lesion, under treatment, was observed in one patient. The finding of active lesions, either in dogs or humans, confirmed the existence of active tegumentary leishmaniasis in Maricá, indicating the need for further studies to evaluate the importance of this infection in the municipality.