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IScience Nov 2023Genetic studies have elucidated the critical roles of Phf7 in germline development in animals; however, the exact etiology of Phf7 mutations leading to male infertility...
Genetic studies have elucidated the critical roles of Phf7 in germline development in animals; however, the exact etiology of Phf7 mutations leading to male infertility and the possibility of mechanism-based therapy are still unclear and warrant further investigation. Using the Phf7 knockout mouse model, we verified that genetic defects were responsible for male infertility by preventing histone-to-protamine exchange, as previously reported. The deficiency of spermatogenesis caused by Phf7 deletion through the endogenous retrovirus-mediated activation of the immune pathway is a common mechanism of infertility. Furthermore, we identified PPARα as a promising target of immunity and inflammation in the testis, where endogenous retroviruses are suppressed, and Phf7 as a crucial regulator of endogenous retrovirus-mediated immune regulation and revealed its role as an epigenetic reader. The loss of Phf7 activates immune pathways, which can be rescued by the PPARα agonist astaxanthin. These results showed that astaxanthin is a potential therapeutic agent for treating male infertility. The findings in our study provide insights into the molecular mechanisms underlying male infertility and suggest potential targets for future research and therapeutic development.
PubMed: 37920670
DOI: 10.1016/j.isci.2023.108030 -
International Journal of Molecular... Oct 2023Small RNA-sequencing (small RNA-seq) has revealed the presence of small RNA-naturally occurring variants such as microRNA (miRNA) isoforms or isomiRs. Due to their small...
Small RNA-sequencing (small RNA-seq) has revealed the presence of small RNA-naturally occurring variants such as microRNA (miRNA) isoforms or isomiRs. Due to their small size and the sequence similarity among miRNA isoforms, their validation by RT-qPCR is challenging. We previously identified two miR-31-5p isomiRs-the canonical and a 3'isomiR variant (3' G addition)-which were differentially expressed between individuals with azoospermia of different origin. Here, we sought to determine the discriminatory capacity between these two closely-related miRNA isoforms of three alternative poly(A) based-RT-qPCR strategies in both synthetic and real biological context. We found that these poly(A) RT-qPCR strategies exhibit a significant cross-reactivity between these miR-31-5p isomiRs which differ by a single nucleotide, compromising the reliable quantification of individual miRNA isoforms. Fortunately, in the biological context, given that the two miRNA variants show changes in the same direction, RT-qPCR results were consistent with the findings of small RNA-seq study. We suggest that miRNA selection for RT-qPCR validation should be performed with care, prioritizing those canonical miRNAs that, in small RNA-seq, show parallel/homogeneous expression behavior with their most prevalent isomiRs, to avoid confounding RT-qPCR-based results. This is suggested as the current best strategy for robust biomarker selection to develop clinically useful tests.
Topics: Humans; MicroRNAs; RNA-Seq; Sequence Analysis, RNA; Base Sequence; Protein Isoforms
PubMed: 37895116
DOI: 10.3390/ijms242015436 -
Basic and Clinical Andrology Oct 2023Sperm banks face a continuously evolving gap between the increasing demand for sperm donation (SD) vs. limited available reserve. To improve donors' recruitment and...
BACKGROUND
Sperm banks face a continuously evolving gap between the increasing demand for sperm donation (SD) vs. limited available reserve. To improve donors' recruitment and increase supply, motivations towards SD should be investigated specifically among young men who have the potential to become donors. Our aim was to evaluate factors which increase and decrease predisposition to donate sperm among non-donor students, who represent a "potential pool" for possible donors' recruitment.
RESULTS
Ninety-three men fulfilled the questionnaire with mean age of 28.2 ± 4.5 years. The most powerful incentive to donate sperm was financial reward followed by a willingness to help others to build a family (3.8 and 3.4, respectively). The most dominant consideration to decline donation was the fear of anonymity loss and future regret (4 and 3.8). While participants' willingness for anonymous SD was fair (2.8), the open-identity donation was rated significantly lower (1.75, p < 0.01). Familiarity with recipients and offspring had lower scores (1.9-2.2) as well.
CONCLUSIONS
Young single men represent a suitable cohort for anonymous donation. Financial reward and willingness to help others are important positive incentives while anonymity preservation is crucial to maintain their willingness towards SD. Regulatory shifting towards open-identity SD necessitates the establishment of an alternative "potential pool" population as a reliable source to recruit donors.
PubMed: 37845630
DOI: 10.1186/s12610-023-00201-2 -
Human Reproduction (Oxford, England) Dec 2023Can in vitro maturation (IVM) and developmental competence of human oocytes be improved by co-culture with ovarian support cells (OSCs) derived from human-induced...
STUDY QUESTION
Can in vitro maturation (IVM) and developmental competence of human oocytes be improved by co-culture with ovarian support cells (OSCs) derived from human-induced pluripotent stem cells (hiPSCs)?
SUMMARY ANSWER
OSC-IVM significantly improves the rates of metaphase II (MII) formation and euploid Day 5 or 6 blastocyst formation, when compared to a commercially available IVM system.
WHAT IS KNOWN ALREADY
IVM has historically shown highly variable performance in maturing oocytes and generating oocytes with strong developmental capacity, while limited studies have shown a positive benefit of primary granulosa cell co-culture for IVM. We recently reported the development of OSCs generated from hiPSCs that recapitulate dynamic ovarian function in vitro.
STUDY DESIGN, SIZE, DURATION
The study was designed as a basic science study, using randomized sibling oocyte specimen allocation. Using pilot study data, a prospective sample size of 20 donors or at least 65 oocytes per condition were used for subsequent experiments. A total of 67 oocyte donors were recruited to undergo abbreviated gonadotropin stimulation with or without hCG triggers and retrieved cumulus-oocyte complexes (COCs) were allocated between the OSC-IVM or control conditions (fetal-like OSC (FOSC)-IVM or media-only IVM) in three independent experimental design formats. The total study duration was 1 April 2022 to 1 July 2023.
PARTICIPANTS/MATERIALS, SETTING, METHODS
Oocyte donors between the ages of 19 and 37 years were recruited for retrieval after informed consent, with assessment of anti-Mullerian hormone, antral follicle count, age, BMI and ovarian pathology used for inclusion and exclusion criteria. In experiment 1, 27 oocyte donors were recruited, in experiment 2, 23 oocyte donors were recruited, and in experiment 3, 17 oocyte donors and 3 sperm donors were recruited. The OSC-IVM culture condition was composed of 100 000 OSCs in suspension culture with hCG, recombinant FSH, androstenedione, and doxycycline supplementation. IVM controls lacked OSCs and contained either the same supplementation, FSH and hCG only (a commercial IVM control), or FOSCs with the same supplementation (Media control). Experiment 1 compared OSC-IVM, FOSC-IVM, and a Media control, while experiments 2 and 3 compared OSC-IVM and a commercial IVM control. Primary endpoints in the first two experiments were the MII formation (i.e. maturation) rate and morphological quality assessment. In the third experiment, the fertilization and embryo formation rates were assessed with genetic testing for aneuploidy and epigenetic quality in blastocysts.
MAIN RESULTS AND THE ROLE OF CHANCE
We observed a statistically significant improvement (∼1.5×) in maturation outcomes for oocytes that underwent IVM with OSCs compared to control Media-IVM and FOSC-IVM in experiment 1. More specifically, the OSC-IVM group yielded a MII formation rate of 68% ± 6.83% SEM versus 46% ± 8.51% SEM in the Media control (P = 0.02592, unpaired t-test). FOSC-IVM yielded a 51% ± 9.23% SEM MII formation rate which did not significantly differ from the media control (P = 0.77 unpaired t-test). Additionally, OSC-IVM yielded a statistically significant ∼1.6× higher average MII formation rate at 68% ± 6.74% when compared to 43% ± 7.90% in the commercially available IVM control condition (P = 0.0349, paired t-test) in experiment 2. Oocyte morphological quality between OSC-IVM and the controls did not significantly differ. In experiment 3, OSC-IVM oocytes demonstrated a statistically significant improvement in Day 5 or 6 euploid blastocyst formation per COC compared to the commercial IVM control (25% ± 7.47% vs 11% ± 3.82%, P = 0.0349 logistic regression). Also in experiment 3, the OSC-treated oocytes generated blastocysts with similar global and germline differentially methylated region epigenetic profiles compared commercial IVM controls or blastocysts after either conventional ovarian stimulation.
LARGE SCALE DATA
N/A.
LIMITATIONS, REASONS FOR CAUTION
While the findings of this study are compelling, the cohort size remains limited and was powered on preliminary pilot studies, and the basic research nature of the study limits generalizability compared to randomized control trials. Additionally, use of hCG-triggered cycles results in a heterogenous oocyte cohort, and potential differences in the underlying maturation state of oocytes pre-IVM may limit or bias findings. Further research is needed to clarify and characterize the precise mechanism of action of the OSC-IVM system. Further research is also needed to establish whether these embryos are capable of implantation and further development, a key indication of their clinical utility.
WIDER IMPLICATIONS OF THE FINDINGS
Together, these findings demonstrate a novel approach to IVM with broad applicability to modern ART practice. The controls used in this study are in line with and have produced similar to findings to those in the literature, and the outcome of this study supports findings from previous co-culture studies that found benefits of primary granulosa cells on IVM outcomes. The OSC-IVM system shows promise as a highly flexible IVM approach that can complement a broad range of stimulation styles and patient populations. Particularly for patients who cannot or prefer not to undergo conventional gonadotropin stimulation, OSC-IVM may present a viable path for obtaining developmentally competent, mature oocytes.
STUDY FUNDING/COMPETING INTEREST(S)
A.D.N., A.B.F., A.G., B.P., C.A., C.C.K., F.B., G.R., K.S.P., K.W., M.M., P.C., S.P., and M.-J.F.-G. are shareholders in the for-profit biotechnology company Gameto Inc. P.R.J.F. declares paid consultancy for Gameto Inc. P.C. also declares paid consultancy for the Scientific Advisory Board for Gameto Inc. D.H.M. has received consulting services from Granata Bio, Sanford Fertility and Reproductive Medicine, Gameto, and Buffalo IVF, and travel support from the Upper Egypt Assisted Reproduction Society. C.C.K., S.P., M.M., A.G., B.P., K.S.P., G.R., and A.D.N. are listed on a patent covering the use of OSCs for IVM: U.S. Provisional Patent Application No. 63/492,210. Additionally, C.C.K. and K.W. are listed on three patents covering the use of OSCs for IVM: U.S. Patent Application No. 17/846,725, U.S Patent Application No. 17/846,845, and International Patent Application No.: PCT/US2023/026012. C.C.K., M.P.S., and P.C. additionally are listed on three patents for the transcription factor-directed production of granulosa-like cells from stem cells: International Patent Application No.: PCT/US2023/065140, U.S. Provisional Application No. 63/326,640, and U.S. Provisional Application No. 63/444,108. The remaining authors have no conflicts of interest to declare.
Topics: Adult; Female; Humans; Male; Young Adult; Coculture Techniques; Follicle Stimulating Hormone; Gonadotropins; In Vitro Oocyte Maturation Techniques; Induced Pluripotent Stem Cells; Oocytes; Pilot Projects; Prospective Studies; Semen
PubMed: 37815487
DOI: 10.1093/humrep/dead205 -
Heliyon Sep 2023Quantum Dots (QDs) modified with branched Polyethylene Glycol-amine (6- or 8-arm PEG-amine) coupled with methoxy PEG (mPEG) hold great promise for biomedical...
Quantum Dots (QDs) modified with branched Polyethylene Glycol-amine (6- or 8-arm PEG-amine) coupled with methoxy PEG (mPEG) hold great promise for biomedical applications due to a long half-life in blood and negligible toxicity. However, the potential risks regarding their concomitant prolonged co-incubation with cardiovascular and blood cells remains inconclusive. In the present study, the feasible, effective and convenient proliferating-restricted cell line models representing the circulatory system were established to investigate the cellular internalization followed by intracellular outcomes and resulting acute/sub-acute cytotoxicity of the 6-arm PEG-amine/mPEG QDs. We found a dose-, time- and cell type-dependent cellular uptake of the 6-arm PEG-amine/mPEG QDs, which was ten-fold lower compared to the traditional linear PEG-modified counterpart. The QDs entered cells via multiple endocytic pathways and were mostly preserved in Golgi apparatus for at least one week instead of degradation in lysosomes, resulting in a minimal acute cytotoxicity, which is much lower than other types of PEG-modified QDs previously reported. However, a sub-acute cytotoxicity of QDs were observed several days post exposure using the concentrations eliciting no-significant acute cytotoxic effects, which was associated with elevated ROS generation caused by QDs remained inside cells. Finally, a non-cytotoxic concentration of the QDs was identified at the sub-acute cytotoxic level. Our study provided important information for clinical translation of branched PEG-amine/mPEG QDs by elucidating the QDs-cell interactions and toxicity mechanism using the proliferation-restricted cell models representing circulatory system. What's more, we emphasized the indispensability of sub-acute cytotoxic effects in the whole biosafety evaluation process of nanomaterials like QDs.
PubMed: 37809902
DOI: 10.1016/j.heliyon.2023.e20028 -
Biology Sep 2023In recent years, there has been an evident delay in childbearing and concerns have been raised about whether this increase in age affects reproductive outcomes. This...
BACKGROUND
In recent years, there has been an evident delay in childbearing and concerns have been raised about whether this increase in age affects reproductive outcomes. This study aimed to evaluate the effect of paternal age on obstetrical and perinatal outcomes in couples undergoing in vitro fertilization or intracytoplasmic sperm injection using autologous sperm and oocytes.
METHODS
This retrospective study evaluated obstetrical and perinatal outcomes from 14,125 couples that were arbitrarily divided into three groups according to paternal age at conception: ≤30 ( = 1164), 31-40 ( = 11,668) and >40 ( = 1293). Statistics consisted of a descriptive analysis followed by univariate and multivariate models, using the youngest age group as a reference.
RESULTS
The study showed significantly longer pregnancies for the fathers aged 31-40 compared to ≤30 years. However, there were no significant differences for the type of delivery, gestational diabetes, anaemia, hypertension, delivery threat, premature rupture of membranes, preterm birth, very preterm birth, and the neonate's sex, weight, low birth weight, very low birth weight, length, cranial perimeter, Apgar score and neonatal intensive care unit admission.
CONCLUSION
Despite our promising results for older fathers, as paternal age was not associated with clinically relevant obstetrical and perinatal outcomes, future well-designed studies are necessary as it has been associated with other important disorders.
PubMed: 37759655
DOI: 10.3390/biology12091256 -
Asian Journal of Andrology Sep 2023Recently discovered microcolonial forms of Mycoplasma hominis (M. hominis) and their impact on human spermatogenesis are studied. The spermatozoa of 125 fertile men...
Recently discovered microcolonial forms of Mycoplasma hominis (M. hominis) and their impact on human spermatogenesis are studied. The spermatozoa of 125 fertile men (sperm donors; from Reprobank [Reproductive Tissue Bank, Moscow, Russia]) and of 93 patients with fertility problems (from the Federal State Budgetary Institution "Research Centre for Medical Genetics [RCMG]", Moscow, Russia) were used. Classical colonies of M. hominis and microcolonies were detected by molecular biological methods, culture of bacteria, and transmission electron microscopy. The unique structure of microcolonial cells, in which the cytoplasmic cylinder is surrounded by concentric electron-dense and electron-light layers with a periodicity of 12-14 nm, and the ability of microcolonial cells to attach to spermatozoa are shown. In patients with lower sperm quality, microcolonies of M. hominis were detected 2.5 times more frequently than classical colonies. The detection of microcolonies in the ejaculate and the frequent isolation of microcolonies from sperm samples of patients with fertility problems suggest that microcolonial cells may be one cause of infertility.
PubMed: 37738137
DOI: 10.4103/aja202333 -
Scientific Reports Sep 2023Sperm quality can be easily influenced by living environmental and occupational factors. This study aimed to discover potential semen quality related living...
Sperm quality can be easily influenced by living environmental and occupational factors. This study aimed to discover potential semen quality related living environmental and occupational factors, expand knowledge of risk factors for semen quality, strengthen men's awareness of protecting their own fertility and assist the clinicians to judge the patient's fertility. 465 men without obese or underweight (18.5 < BMI < 28.5 kg/m), long-term medical history and history of drug use, were recruited between June 2020 to July 2021, they are in reproductive age (25 < age < 45 years). We have collected their semen analysis results and clinical information. Logistic regression was applied to evaluate the association of semen quality with different factors. We found that living environment close to high voltage line (283.4 × 10/ml vs 219.8 × 10/ml, Cohen d = 0.116, P = 0.030) and substation (309.1 × 10/ml vs 222.4 × 10/ml, Cohen d = 0.085, P = 0.015) will influence sperm count. Experienced decoration in the past 6 months was a significant factor to sperm count (194.2 × 10/ml vs 261.0 × 10/ml, Cohen d = 0.120, P = 0.025). Living close to chemical plant will affect semen PH (7.5 vs 7.2, Cohen d = 0.181, P = 0.001). Domicile close to a power distribution room will affect progressive sperm motility (37.0% vs 34.0%, F = 4.773, Cohen d = 0.033, P = 0.030). Using computers will affect both progressive motility sperm (36.0% vs 28.1%, t = 2.762, Cohen d = 0.033, P = 0.006) and sperm total motility (57.0% vs 41.0%, Cohen d = 0.178, P = 0.009). After adjust for potential confounding factors (age and BMI), our regression model reveals that living close to high voltage line is a risk factor for sperm concentration (Adjusted OR 4.03, 95% CI 1.15-14.18, R = 0.048, P = 0.030), living close to Chemical plants is a protective factor for sperm concentration (Adjusted OR 0.15, 95% CI 0.05-0.46, R = 0.048, P = 0.001) and total sperm count (Adjusted OR 0.36, 95% CI 0.13-0.99, R = 0.026, P = 0.049). Time spends on computer will affect sperm total motility (Adjusted OR 2.29, 95% CI 1.11-4.73, R = 0.041, P = 0.025). Sum up, our results suggested that computer using, living and working surroundings (voltage line, substation and chemical plants, transformer room), and housing decoration may association with low semen quality. Suggesting that some easily ignored factors may affect male reproductive ability. Couples trying to become pregnant should try to avoid exposure to associated risk factors. The specific mechanism of risk factors affecting male reproductive ability remains to be elucidated.
Topics: Humans; Male; Middle Aged; Cross-Sectional Studies; East Asian People; Semen; Semen Analysis; Sperm Motility; Adult; Risk Factors; Working Conditions; Neighborhood Characteristics; Fertility; Social Determinants of Health
PubMed: 37735181
DOI: 10.1038/s41598-023-42927-z -
Cureus Aug 2023Background Infertility is a public health issue with a significant impact on the well-being of affected couples. Aim This paper aims to detect the determinants of...
Background Infertility is a public health issue with a significant impact on the well-being of affected couples. Aim This paper aims to detect the determinants of pregnancy and their outcome after assisted reproductive therapy (ART) in a sample of Palestinian society. Methods A retrospective observational study was carried out at Razan Medical Center for Infertility. Subjects were assigned into twelve categories based on the type of infertility (primary versus secondary), the cause of infertility, and the treatment modality. Age at marriage, age at presentation, duration of infertility, in addition to regularity of menstruation, were also studied. Biochemical pregnancy was considered the endpoint for the purpose of the analysis. Results We reviewed the files of 459 subjects diagnosed with infertility. 79.74% had primary infertility while 20.26% had secondary. 28.85% were found to be infertile due to anovulation, 2.86% due to endometriosis, 16.74% attributed to male factor, and 3.3% had tubal damage. 13.43% of cases were multifactorial while 34.80% were idiopathic. Four biochemical markers were assessed in our study: thyroid-stimulating hormone (TSH) (=2.32±2.46), luteinizing hormone (=6.71±4.90), follicle-stimulating hormone (FSH) (=6.59±6.11), and human prolactin (=41.88±6.50). The menstrual cycle was regular in 70.58% of subjects, in contrast to 29.41% with irregular cycles. The female mean age at presentation was 22.76±5.58, while their mean age of marriage was 22.76±4. The mean duration of infertility was 3.97±6.87 years. Patients were treated via three modalities - ovarian stimulation (2.2%), intrauterine insemination (29.58%), and intracytoplasmic sperm injection (68.21%) - with a success rate of 51.85%. Of the studied factors, only diagnosis, follicle-stimulating hormone, and treatment modality had a significant impact on the outcome, with values of 0.040, 0.003, and <.0.001, respectively. Conclusions There is a strong relationship between diagnosis prior to intervention, level of FSH, and treatment modality and successful outcome of ART.
PubMed: 37674937
DOI: 10.7759/cureus.43011 -
Frontiers in Cell and Developmental... 2023Advances in cancer treatments have determined an increase in survival rates. However, these lifesaving therapies may have a negative impact on reproductive health. To...
Advances in cancer treatments have determined an increase in survival rates. However, these lifesaving therapies may have a negative impact on reproductive health. To diminish the infertility risk; different fertility preservation strategies have been designed. Sperm freezing is the gold standard fertility preservation method in the case of post-pubertal men. The main objective of this study is to evaluate the fertility status of Uruguayan male cancer survivors who have gone through sperm freezing, as well as to assess oncofertility counseling received by these patients. This is a descriptive, cross-sectional, observational, and transversal study. A survey was conducted on male cancer survivors who cryopreserved sperm between 1985 and 2021 in "Reprovita Lab and Biobank" which is the only sperm bank in this country. One hundred thirty-five participants answered the survey. At the time of diagnosis, the mean age of patients was 28.8 ± 6.4 years old. Testicular was the most frequent type of cancer (64%). Only, 12% ( = 15) already had children at the time of diagnosis. Among the interviewed survivors, 50% ( = 62) attempted to conceive after cancer treatment, and 68% ( = 42) achieved natural pregnancy. Patients who did not achieve spontaneous conception ( = 11), used their cryopreserved samples, and 45.4% achieved pregnancy. About 86% ( = 107) of survivors believed that the timing of oncofertility referrals was appropriate and 97% considered that having the possibility of protecting their fertility was very important. Eighty percent ( = 101), were advised by their attending physicians, 14% ( = 18) sought advice from family members or friends, and 4% ( = 5) from oncofertility specialists. To our knowledge, this is the first study evaluating the reproductive outcomes of male cancer survivors in our country and the region. Most of the interviewed survivors considered fertility preservation as a positive initiative, independent of their reproductive outcomes, reflecting the importance of fertility preservation counseling as one of the most important aspects for futurequality of life of young cancer patients.
PubMed: 37664467
DOI: 10.3389/fcell.2023.1240152