An investigation of racial and ethnic disparities in donor sperm availability in the United States.

OBJECTIVE: To determine whether the racial and ethnic distribution of sperm donors contributing to US sperm banks differs from the demographics of the US population and those of US donor sperm recipients undergoing treatment with in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). DESIGN: Cross-sectional study. SETTING: United States donor sperm banks, US Census, and fertility clinics reporting to the Society for Assisted Reproductive Technology Clinic Outcome Reporting System. PATIENTS: Sperm donors from 18 banks, men aged 18-39 years on the basis of the 2021 Census Current Population Survey, and recipients of donor sperm for IVF and ICSI treatments in clinics reporting to the Assisted Reproductive Technology Clinic Outcome Reporting System from January 1, 2018, to December 31, 2020. INTERVENTION: None. MAIN OUTCOME MEASURE: Proportions of sperm donors and donor sperm recipients undergoing IVF and ICSI treatments identifying as each racial and ethnic group. RESULTS: Eighteen donor sperm banks were identified, encompassing 1,503 donors. Of these donors, 60.9% identified as White compared with 55.1% of the US male population and 67.7% of donor sperm recipients. Proportions of donors identifying as Asian or 2 or more races were larger than those of US men and donor sperm recipients (Asian: 18.6% vs. 6.5%, and 18.6% vs. 10.2%; 2 or more races: 11.6% vs. 2.2%, and 11.6% vs. 1.7%). In contrast, Black donors were underrepresented when compared with the US population of men and donor sperm recipients (2.8% vs. 12.9% and 2.8% vs. 13.0%). Hispanic donors were underrepresented when compared with the US population of men (6.0% vs. 22.2%). The percentages of Hispanic sperm donors and donor sperm recipients were similar (6.0% vs. 7.0%). CONCLUSION: The racial and ethnic distribution of sperm donors differs significantly from the demographics of the US male population and donor sperm recipients undergoing IVF and ICSI treatments. These findings suggest a need for targeted recruitment efforts for Black sperm donors.

Addressing Privacy Concerns Surrounding Oocyte Donation in the United States: Gone With Anonymity.

Oocyte donation has greatly expanded in the past several decades since the first procedure was performed in 1983. According to the Centers for Disease Control, the number of cycles using donor oocytes increased from 18,011 cycles in 2010 to 27,131 cycles in 2019. Oocyte donation has become an important reproductive option for women with diminished ovarian reserve, recurrent failed in vitro fertilization, or heritable genetic conditions. It is also particularly important for single men, same-sex male couples, and men with a transgender woman partner. More recently, societal changes accompanying the expansion of social media and broader access to direct-to-consumer DNA testing have raised concerns about privacy and anonymity. In this article, we review two specific aspects of donor privacy: privacy related to personal identifiers provided by clinics or donor egg bank websites and privacy related to direct-to-consumer genetic testing. We also provide clinical recommendations specific to the United States for working with oocyte donors and donor oocyte recipients.

"Can't Touch This!": Laparoscopic management of an obstructed uterus didelphys before and after treatment for pelvic inflammatory disease.

OBJECTIVE: To describe the laparoscopic management of an obstructed uterus didelphys before and after treatment for pelvic inflammatory disease. To compare the appearance of pelvic organs during active infection with their appearance after washout and appropriate antibiotic treatment, emphasizing the importance of knowing when to abort a procedure. DESIGN: Video demonstration of surgical and medical management considerations during a complex pelvic surgery. Visualization of tissue healing that occurs with appropriate antibiotic treatment. SETTING: Academic Center. PATIENT: A patient who presents for definitive surgical management of a uterus didelphys with an obstruction at her right hemicervix. Her presentation is complicated by a tubo-ovarian abscess. INTERVENTION: A uterus didelphys is classically defined as two hemiuteri with duplicated cervices with or without a longitudinal vaginal septum. Uterus didelphys may have an obstruction and/or communication between the two uterine horns, in which case patients may present with complications such as cyclic pelvic pain from hematometra or genital tract infection. This is a case report of a 14-year-old G0 who presented to the emergency department with two weeks of vaginal bleeding, severe diffuse abdominal pain, and malodorous vaginal discharge. Transabdominal ultrasound and a magnetic resonance imaging of the pelvis established a new diagnosis of a uterus didelphys with an obstruction at her right hemicervix and a fistulous tract connecting her right and left hemiuteri at the level of the internal cervical os. She was also found to have a 3 cm left ovarian cyst and a new finding of congenital absence of her right kidney. Patient was administered ceftriaxone, doxycycline, and metronidazole antibiotics as treatment of presumed pelvic inflammatory disease but experienced minimal improvement after 24 hours. The decision was made to proceed with surgical intervention. A survey of the pelvis revealed significant inflammation, friable peritoneum, and endometriosis. The uterine horns in didelphic configurations were visualized. The fimbriae at the left fallopian tube were notably splayed out, swollen, and inflamed. There was a notable large mass in the location where the ovarian cyst had been previously described on imaging. A large amount of purulent material was expressed when compressed, consistent with a tubo-ovarian abscess. The infection likely originated from the menstrual blood collection at the right obstructed cervix that ascended through the communication between the right and left hemiuteri. The pelvis was irrigated thoroughly. At this point, the decision was made to stop the procedure, pursue antibiotic treatment, and resolve the active infection before correcting her complex müllerian anomaly. Patient continued on her antibiotic course, which included piperacillin-tazobactam, while hospitalized, followed by a five-day course of amoxicillin-clavulanate. She was also placed on medroxyprogesterone acetate for menstrual suppression. MAIN OUTCOME MEASURE: Advantage of allowing time for antibiotic treatment and tissue healing before repair of a complex müllerian anomaly. RESULT: With antibiotic treatment, she recovered well postoperatively with resolution of her pain. Three months later, she returned to the operating room for definitive surgical management of her obstructed uterine didelphys. On laparoscopy, there was a significant improvement in tissue quality. Most notably, the fimbriae of the left fallopian tube were no longer inflamed. We proceeded with the planned correction of the complex müllerian anomaly. After resection of the right uterine horn, the fistula tract was identified and also resected. The defect in the right hemicervix was closed over, reinforcing the medial side of the left hemicervix. She had an uncomplicated postoperative recovery, and menses resumed without pain. CONCLUSIONS: The presented case provides unique insight into the tissue healing that occurs before and after antibiotic treatment. Knowing when to stop, especially in the setting of an active infection, is extremely important for performing a procedure safely, minimizing harm, and allowing for robust tissue repair. It is also important to optimize modifiable preoperative factors before correcting a complex müllerian anomaly. Assessing and reassessing the situation during a complex pelvic surgery is essential, especially in the setting of a complex müllerian anomaly where the preoperative examination and imaging may not be definitive.

Embryo ploidy in vitrified versus fresh oocytes: Is there a difference?

PURPOSE: To evaluate embryo ploidy in a cohort of patients who underwent preimplantation genetic testing for aneuploidy (PGT-A) with vitrified oocytes compared to fresh oocytes. METHODS: Patients who underwent their first autologous oocyte vitrification and warming followed by in vitro fertilization (IVF) and trophectoderm biopsy for PGT-A between 1/1/2017 and 12/31/2021 at a single academic institution were included. Patients were compared 1:3 to age-matched controls who underwent their first IVF cycle with fresh oocytes and subsequent trophectoderm biopsy for PGT-A. The primary outcome was the proportions of euploid, mosaic, and aneuploid embryos between those using vitrified versus fresh oocytes. RESULTS: 117 patients who cryopreserved a total of 1,272 mature oocytes were included in the study and were matched with 351 controls using fresh oocytes. The average age was 36.9 ± 2.6 years, and the median interval between oocyte vitrification and warming was 38 months. There were similar numbers of mature oocytes (10.9 ± 4.9 vs. 11.1 ± 6.3, P = .67), fertilized oocytes (7.8 ± 4.0 vs. 8.7 ± 5.5, P = .10), and blastocysts per patient (5.1 ± 3.1 vs. 5.8 ± 4.3, P = .10) between those using vitrified versus fresh oocytes. In terms of embryo ploidy results, there were no statistically significant differences in rates of euploidy (40.1% vs. 41.6%), mosaicism (15.7% vs. 12.0%), or aneuploidy (44.3% vs. 46.4%) (P = .06) between the two groups. CONCLUSIONS: Oocyte vitrification with subsequent warming, fertilization, and trophectoderm biopsy for PGT-A was not associated with adverse chromosomal competence when compared to age-matched controls utilizing fresh oocytes.

Association between donor gamete use and supernumerary embryo disposition decisions.

PURPOSE: The aim of this study is to determine if donor gamete use is associated with patients' decisions regarding disposition of supernumerary embryos. METHODS: Patients who intended to undergo an IVF cycle at a single academic center signed an embryo disposition consent form to indicate their disposition preferences for any supernumerary embryos. A retrospective chart review was performed to obtain the embryo disposition declarations and demographic information. The primary outcome was the distribution of embryo disposition choices between patients who used donor gametes compared to patients who did not use donor gametes. Fisher's exact test was used to compare groups. Logistic regression models were created to determine the association between donor gamete use and disposition decision after adjusting for patient age, body mass index, and nulliparity. RESULTS: Five hundred six patients were included. Ninety-one (18.0%) patients used donor gametes [46 (9.0%) donor oocytes, 52 (10.3%) donor sperm]. Patients using donor gametes differed from those not using donor gametes when making decisions concerning death of the patient (P < 0.01), simultaneous death (P = 0.04), separation (P < 0.01), discontinuation of ART (P = 0.01), and time-limited storage (P < 0.01). Most patients, regardless of donor or autologous gamete use, awarded embryos to themselves or their partner if given the option. For patients who did not choose this option, excess embryos were generally awarded to research or discarded rather than donating to another couple. Patients using donor gametes were more likely to award embryos to research over discarding. CONCLUSION: Patients using donor gametes made different choices regarding supernumerary embryo disposition compared to patients not using donor gametes.

Ovarian Reserve Disorders, Can We Prevent Them? A Review.

The ovarian reserve is finite and begins declining from its peak at mid-gestation until only residual follicles remain as women approach menopause. Reduced ovarian reserve, or its extreme form, premature ovarian insufficiency, stems from multiple factors, including developmental, genetic, environmental exposures, autoimmune disease, or medical/surgical treatment. In many cases, the cause remains unknown and resulting infertility is not ultimately addressed by assisted reproductive technologies. Deciphering the mechanisms that underlie disorders of ovarian reserve could improve the outcomes for patients struggling with infertility, but these disorders are diverse and can be categorized in multiple ways. In this review, we will explore the topic from a perspective that emphasizes the prevention or mitigation of ovarian damage. The most desirable mode of fertoprotection is primary prevention (intervening before ablative influence occurs), as identifying toxic influences and deciphering the mechanisms by which they exert their effect can reduce or eliminate exposure and damage. Secondary prevention in the form of screening is not recommended broadly. Nevertheless, in some instances where a known genetic background exists in discrete families, screening is advised. As part of prenatal care, screening panels include some genetic diseases that can lead to infertility or subfertility. In these patients, early diagnosis could enable fertility preservation or changes in family-building plans. Finally, Tertiary Prevention (managing disease post-diagnosis) is critical. Reduced ovarian reserve has a major influence on physiology beyond fertility, including delayed/absent puberty or premature menopause. In these instances, proper diagnosis and medical therapy can reduce adverse effects. Here, we elaborate on these modes of prevention as well as proposed mechanisms that underlie ovarian reserve disorders.

Fertility awareness and attitudes among resident physicians across different specialties.

PURPOSE: To evaluate knowledge of age-related fertility decline and oocyte cryopreservation among resident physicians in obstetrics and gynecology (ob-gyn) compared to residents in other specialties. METHODS: An online survey was sent to the US residency program directors for ob-gyn, internal medicine, emergency medicine, family medicine, general surgery, pediatrics, and psychiatry. They were asked to forward the survey to their respective residents. The survey consisted of three sections: fertility knowledge, oocyte cryopreservation knowledge, and attitudes toward family building and fertility preservation. Multivariable logistic regression models were used to compare outcomes between ob-gyn and non-ob-gyn residents. RESULTS: Of the 2,828 completed surveys, 450 (15.9%) were by ob-gyn residents and 2,378 (84.1%) were by residents in other specialties. 66.3% of respondents were female. The median number of correct answers was 2 out of 5 on the fertility knowledge section and 1 out of 3 on the oocyte cryopreservation knowledge section among both ob-gyn and non-ob-gyn residents. After adjusting for covariates, residents in ob-gyn were no more likely to answer these questions correctly than residents in other specialties (fertility knowledge, adjusted OR .97, 95% CI .88-1.08; oocyte cryopreservation knowledge, adjusted OR 1.05, 95% CI .92-1.19). Ob-gyn residents were significantly more likely than non-ob-gyn residents to feel "somewhat supported" or "very supported" by their program to pursue family building goals (83.5% vs. 75.8%, OR 1.62, 95% CI 1.23-2.14). CONCLUSIONS: Resident physicians, regardless of specialty, have limited knowledge of natural fertility decline and the opportunity to cryopreserve oocytes. These data suggest need for improved fertility education.

Racial and ethnic disparities among donor oocyte banks in the United States.

OBJECTIVE: To determine whether racial and ethnic distributions of oocyte donors contributing to US oocyte banks differ from the demographics of US women and donor oocyte recipients. DESIGN: Cross-sectional study. SETTING: United States donor oocyte banks, US census, and fertility clinics reporting to the Society for Assisted Reproductive Technology Clinic Outcome Reporting System. PATIENTS: Oocyte donors from 12 banks, women aged 18-44 years based on the 2019 census, and US recipients of cryopreserved donor oocytes from 2012 to 2015. INTERVENTION: None. MAIN OUTCOME MEASURE: Proportions of donors identifying as each racial and ethnic group. RESULTS: Of the 1,574 oocyte donors, 678 (43.1%) identified as white compared with 54.8% of US women and 69.1% of donor oocyte recipients. Proportions of donors identifying as Hispanic or two or more races were larger than those of US women and donor oocyte recipients (Hispanic: 24.1% vs. 20.8%, and 24.1% vs. 8.8%, respectively; two or more races: 16.1% vs. 2.3%, and 16.1% vs. 0.5%, respectively). African American donors were underrepresented compared with US women (8.9% vs. 14.0%) and oocyte recipients (8.9% vs. 10.8%). Although the proportion of Asian donors was similar to that of US women (7.7% vs. 7.1%), Asian donors were underrepresented compared with donor oocyte recipients (7.7% vs. 10.6%). CONCLUSION: Racial and ethnic distribution of oocyte donors differs significantly from the demographics of US women and cryopreserved donor oocyte recipients. These data suggest a need for targeted recruitment of African American and Asian oocyte donors.

Spaced Repetition Flashcards for Teaching Medical Students Psychiatry.

OBJECTIVE: Retrieval practice, often using electronic flashcards, is increasingly utilized among medical students for self-study. In this study, the authors evaluated usage and satisfaction with electronic flashcards based on a medical school psychiatry curriculum. METHODS: First-year medical students at one institution consented to participate and received access to a set of pre-made flashcards. Surveys were distributed that collected demographic information along with measures of prior performance, test anxiety, and prior experience with electronic flashcards. The total number of flashcard reviews and time spent on the platform for each student were collected using statistics internally generated by the platform. Each student's final exam score was also collected. RESULTS: A total of 114 of 129 first year medical students (88%) consented to participate, and 101 students were included in the final analysis. Fifty-eight (56%) were flashcard users with a median of 660 flashcards studied over 2.95 h. A total of 87% of flashcard users found the flashcards to be helpful, and 83% of flashcard users would recommend the flashcards to someone else. Flashcard usage was not associated with final exam scores. CONCLUSIONS: This novel electronic study resource was well-received by first-year medical students for psychiatric instruction in medical school, though usage was not associated with higher exam scores.

Novel Spaced Repetition Flashcard System for the In-training Examination for Obstetrics and Gynecology.

INTRODUCTION: Electronic flashcards allow repeated information exposure over time along with active recall. It is increasingly used for self-study by medical students but remains poorly implemented for graduate medical education. The primary goal of this study was to determine whether a flashcard system enhances preparation for the in-training examination in obstetrics and gynecology (ob-gyn) conducted by the Council on Resident Education in Obstetrics and Gynecology (CREOG). METHODS: Ob-gyn residents at Duke University were included in this study. A total of 883 electronic flashcards were created and distributed. CREOG scores and flashcard usage statistics, generated internally by interacting with the electronic flashcard system, were collected after the 2019 exam. The primary outcome was study aid usage and satisfaction. The secondary outcome was the impact of flashcard usage on CREOG exam scores. RESULTS: Of the 32 residents, 31 (97%) participated in this study. Eighteen (58%) residents used the study's flashcards with a median of 276 flashcards studied over a median of 3.7 h. All of the flashcard users found the study aid helpful, and all would recommend them to another ob-gyn resident. Using the flashcards to study for the 2019 CREOG exam appeared to correlate with improvement in scores from 2018 to 2019, but did not achieve statistical significance after adjusting for post-graduate year (beta coefficient = 10.5; 95% confidence interval = - 0.60,21.7; p = 0.06). DISCUSSION: This flashcard resource was well received by ob-gyn residents for in-training examination preparation, though it was not significantly correlated with improvement in CREOG scores after adjusting for post-graduate year.

Correction to: Novel Spaced Repetition Flashcard System for the In-training Examination for Obstetrics and Gynecology.

[This corrects the article DOI: 10.1007/s40670-021-01320-z.].

Surrogacy Laws in the United States: What Obstetrician-Gynecologists Need to Know.

The first child carried by a surrogate after in vitro fertilization in the United States was born in 1985. Since then, the number of such births has steadily grown. According to the Centers for Disease Control and Prevention, the number of gestational carrier cycles increased from 727 in 1999 to 3,432 in 2013, encompassing more than 18,000 children born over this period. Surrogacy offers an alternative to adoption. However, it also disrupts traditional notions of parentage and gestation and complicates the role of obstetrician-gynecologists (ob-gyns) in helping their patients navigate difficult ethical issues. Surrogacy legislation falls under the jurisdiction of each individual state, which results in a variety of approaches. In this article, we review the legal aspects of surrogacy important for specialist ob-gyns, including select landmark court cases, states' approaches to surrogacy legislation, and unique components of informed consent. We also provide clinical recommendations specific to the United States for working with gestational surrogates and intended parents, spanning preconception, prenatal care, and delivery.

Syphilis in Pregnancy.

IMPORTANCE: Since 2013, the United States has seen a rise in cases of congenital syphilis, culminating in a relative increase of 153% from 2013 to 2017 and 918 reported cases in 2017. In all, 50% to 80% of pregnant women with syphilis experience an adverse pregnancy outcome including stillbirth or spontaneous abortion. OBJECTIVE: This article aims to review the current evidence and recommendations for management of syphilis in pregnancy. EVIDENCE ACQUISITION: Original research articles, review articles, and guidelines on syphilis were reviewed. RESULTS: In pregnancy, routine screening for syphilis is recommended on initiation of prenatal care. In high-risk populations, repeat testing is recommended in the early third trimester and at delivery. Penicillin remains the recommended treatment in pregnancy. After treatment, nontreponemal titers should be repeated at minimum during the early third trimester and at delivery to assess for serologic response. In high-risk populations, titers should be repeated monthly. CONCLUSION AND RELEVANCE: Routine screening in pregnancy is essential for identification of syphilis infection and prevention of congenital syphilis. Subsequent adequate treatment with penicillin therapy more than 30 days before delivery and at the correct dosages depending on the stage of infection should be incorporated into clinical practice.

Approval-adjusted recall rates of high-risk medical devices from 2002-2016 across food and drug administration device categories.

Between 2002 and 2016, 806 million medical devices were recalled. When approving a device, the FDA employs advisory boards organized by medical specialty (e.g. cardiovascular) to make approval recommendations. Previous work has demonstrated high numbers of recalled orthopedic and cardiovascular devices; however, no prior studies have controlled for the number of approvals by advisory board. The purpose of this study is to identify device fields at higher risk for safety problems. This study compares specialty-specific, approval-adjusted recall rates of high-risk medical devices from 2002 to 2016 by utilizing publicly available FDA data on recalls and approvals. Devices approved under general hospital (113), anesthesiology (98), and cardiovascular (98) advisory boards constituted 71% of all class I recalls. For devices approved via the more rigorous pre-market approval pathway, those under the purview of the general hospital (0.25 recalls/approval, 95% CI 0.15 - 0.41) advisory board had a significantly higher rate than average (p<0.05). For 510(k) cleared devices, microbiology (6.0 recalls/clearance, 95% CI 3.4 - 10.6), anesthesiology (0.04 recalls/clearance, 95% CI 0.03 - 0.04), general hospital (0.02 recalls/clearance, 95% CI 0.02 - 0.02), and cardiovascular (0.010 recalls/ clearance, 0.009 to 0.015) advisory boards had significantly higher recall rates than average (p<0.05). Future regulatory resources should be directed towards device areas and approval pathways that pose a higher risk for safety problems.

Medical Device Approvals Through the Premarket Approval Pathway in Obstetrics and Gynecology From 2000 to 2015: Process and Problems.

Recent controversies surrounding obstetrics and gynecology devices, including a permanent sterilization device, pelvic meshes, and laparoscopic morcellators, highlight the need for deeper understanding of obstetrics and gynecology medical device regulation. The U.S. Food and Drug Administration premarket approval database was queried for approvals assigned to the obstetrics and gynecology advisory committee from January 2000 to December 2015. Eighteen device approvals occurred in the time period studied. The most common clinical indications included endometrial ablation (33%), contraception (28%), and fetal monitoring (17%). The median approval time was 290 days (range 178-1,399 days). Regarding the pivotal trials leading to approval, there were 11 randomized controlled trials, one randomized crossover study, five nonrandomized prospective studies, and two human factor studies. Fourteen devices (78%) met their primary clinical efficacy endpoint. Only 12 of 18 devices were required to conduct postmarket surveillance. A significant proportion of devices (42%) were approved on the basis of nonrandomized controlled trials. Three devices have been withdrawn after approval, all of which were either not referred or not recommended for approval by the obstetrics and gynecology advisory committee. Of the three devices withdrawn from the market, two failed to demonstrate clinical benefit in their pivotal trials. One device was not required to undergo postmarketing surveillance and was subsequently withdrawn as a result of patient safety concerns. Our results reveal significant weaknesses in the preapproval and postapproval regulation of high-risk obstetrics and gynecology devices. Greater specialty group involvement is necessary to ensure the development of safe and clinically effective devices.

Biochemical and cellular analysis of human variants of the DYT1 dystonia protein, TorsinA/TOR1A.

Early-onset dystonia is associated with the deletion of one of a pair of glutamic acid residues (c.904_906delGAG/c.907_909delGAG; p.Glu302del/Glu303del; ΔE 302/303) near the carboxyl-terminus of torsinA, a member of the AAA(+) protein family that localizes to the endoplasmic reticulum lumen and nuclear envelope. This deletion commonly underlies early-onset DYT1 dystonia. While the role of the disease-causing mutation, torsinAΔE, has been established through genetic association studies, it is much less clear whether other rare human variants of torsinA are pathogenic. Two missense variations have been described in single patients: R288Q (c.863G>A; p.Arg288Gln; R288Q) identified in a patient with onset of severe generalized dystonia and myoclonus since infancy and F205I (c.613T>A, p.Phe205Ile; F205I) in a psychiatric patient with late-onset focal dystonia. In this study, we have undertaken a series of analyses comparing the biochemical and cellular effects of these rare variants to torsinAΔE and wild-type (wt) torsinA to reveal whether there are common dysfunctional features. The results revealed that the variants, R288Q and F205I, are more similar in their properties to torsinAΔE protein than to torsinAwt. These findings provide functional evidence for the potential pathogenic nature of these rare sequence variants in the TOR1A gene, thus implicating these pathologies in the development of dystonia.

Dopa-responsive dystonia: functional analysis of single nucleotide substitutions within the 5' untranslated GCH1 region.

BACKGROUND: Mutations in the GCH1 gene are associated with childhood onset, dopa-responsive dystonia (DRD). Correct diagnosis of DRD is crucial, given the potential for complete recovery once treated with L-dopa. The majority of DRD associated mutations lie within the coding region of the GCH1 gene, but three additional single nucleotide sequence substitutions have been reported within the 5' untranslated (5'UTR) region of the mRNA. The biologic significance of these 5'UTR GCH1 sequence substitutions has not been analyzed. METHODOLOGY/PRINCIPAL FINDINGS: Luciferase reporter assays, quantitative real time PCR and RNA decay assays, combined with bioinformatics, revealed a pathogenic 5'UTR GCH1 substitution. The +142C>T single nucleotide 5'UTR substitution that segregates with affected status in DRD patients, substantially attenuates translation without altering RNA expression levels or stability. The +142C>T substitution disrupts translation most likely by creating an upstream initiation start codon (uAUG) and an upstream open reading frame (uORF). CONCLUSIONS/SIGNIFICANCE: This is the first GCH1 regulatory substitution reported to act at a post-transcriptional level, increasing the list of genetic diseases caused by abnormal translation and reaffirming the importance of investigating potential regulatory substitutions in genetic diseases.