Euploid programmed frozen embryo transfer cycles are associated with a higher live birth rate when estradiol levels more closely mimic physiology.

OBJECTIVE: To determine whether peak estradiol (E2) levels above the usual physiologic range (300-500 pg/mL) will impact programmed frozen embryo transfer (FET) outcomes in an ideal study population of those using good-quality single euploid blastocysts. DESIGN: Retrospective cohort study. SETTING: University-based clinic. PATIENTS: Single euploid-programmed FET done at a single academic institution from January 2016 to December 2019. The population was divided into three groups on the basis of peak serum E2 levels during endometrial preparation: group A (E2 <300 pg/mL), group B (300-500 pg/mL), and group C (>500 pg/mL). Group B was used as the reference range for statistical analysis. INTERVENTION: Frozen embryo transfer cycles. MAIN OUTCOME MEASURES: The primary outcome was the live birth rate (LBR). Secondary outcomes included implantation, biochemical, ectopic, and miscarriage rates. RESULTS: A total of 750 FET cycles were included in this study. Poisson regression analysis showed a negative impact of higher peak E2 on the LBR. A decrease in LBR was noted between group C and referent group B (50.2% vs. 63.4%, risk ratio 0.79 [0.68-0.91]) and group A and referent group B (42.5% vs. 63.4%, risk ratio 0.67 [0.46-0.98]). Secondary outcomes were notable for a lower implantation rate when groups A and C were compared with group B and a higher biochemical rate between group C and group B. There was no notable difference between groups in ectopic or miscarriage rates. CONCLUSION: Limiting peak serum E2 levels to 300-500 pg/mL during programmed FET cycles is associated with improved LBRs compared with cycles with peak E2 levels of <300 pg/mL or >500 pg/mL in an ideal study population.

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.

Cavitation in a periodontal pocket by an ultrasonic dental scaler: A numerical investigation.

Periodontal pockets are spaces or holes surrounded by teeth under the gum line. These pockets can become filled with infection-causing bacteria resulting in tissue, bone, and tooth loss. Cavitation produced by the oscillating tip of dental ultrasonic scalers plays a significant role in routine periodontal therapy to clean these areas. Numerical studies were conducted for a scaler vibrating in a periodontal pocket which was simplified to a hole, using ABAQUS based on the finite element method. The simulations consider the three-dimensional, nonlinear, and transient interaction between the vibration and deformation of the scaler tip, the water flow around the scaler and the cavitation formation. The numerical model was validated by comparing results with experimental data for a scaler vibrating in an unbounded liquid, the displacement at the free end of the scaler and the cavitation pattern near the scaler tip displaying excellent agreement. A parametric study for a scaler vibrating in a hole has been carried out in terms of the volume of the hole, the taper ratio (the radius ratio between the circular opening and bottom of the hole), and the immersion depth of the scaler tip in the hole. The amount of cavitation generated is evaluated by the cavitation density (or the void fraction) which is the ratio of the volume of the cavitation occupied in the hole to the total volume of the hole. Numerical results indicate that the cavitation density in the hole increases with the decreasing hole volume and the increasing taper ratio. It is inferred that cleaning effects could be increased if some modifications to the scaler design could be made to increase the blocking effect of the hole during the cleaning process. Cavitation is observed in the hole even if the scaler is placed above the hole and increases with the immersion depth.

Contour, a semi-automated segmentation and quantitation tool for cryo-soft-X-ray tomography.

Cryo-soft-X-ray tomography is being increasingly used in biological research to study the morphology of cellular compartments and how they change in response to different stimuli, such as viral infections. Segmentation of these compartments is limited by time-consuming manual tools or machine learning algorithms that require extensive time and effort to train. Here we describe Contour, a new, easy-to-use, highly automated segmentation tool that enables accelerated segmentation of tomograms to delineate distinct cellular compartments. Using Contour, cellular structures can be segmented based on their projection intensity and geometrical width by applying a threshold range to the image and excluding noise smaller in width than the cellular compartments of interest. This method is less laborious and less prone to errors from human judgement than current tools that require features to be manually traced, and does not require training datasets as would machine-learning driven segmentation. We show that high-contrast compartments such as mitochondria, lipid droplets, and features at the cell surface can be easily segmented with this technique in the context of investigating herpes simplex virus 1 infection. Contour can extract geometric measurements from 3D segmented volumes, providing a new method to quantitate cryo-soft-X-ray tomography data. Contour can be freely downloaded at github.com/kamallouisnahas/Contour.

Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells.

Imaging of actin filaments is crucial due to the integral role that they play in many cellular functions such as intracellular transport, membrane remodelling and cell motility. Visualizing actin filaments has so far relied on fluorescence microscopy and electron microscopy/tomography. The former lacks the capacity to capture the overall local ultrastructure, while the latter requires rigorous sample preparation that can lead to potential artefacts, and only delivers relatively small volumes of imaging data at the thinnest areas of a cell. In this work, a correlative approach utilizing in situ super-resolution fluorescence imaging and cryo X-ray tomography was used to image bundles of actin filaments deep inside cells under near-native conditions. In this case, fluorescence 3D imaging localized the actin bundles within the intracellular space, while X-ray tomograms of the same areas provided detailed views of the local ultrastructure. Using this new approach, actin trails connecting vesicles in the perinuclear area and hotspots of actin presence within and around multivesicular bodies were observed. The characteristic prevalence of filamentous actin in cytoplasmic extensions was also documented.

Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells.

Three-dimensional (3D) structured illumination microscopy (SIM) allows imaging of fluorescently labelled cellular structures at higher resolution than conventional fluorescence microscopy. This super-resolution (SR) technique enables visualization of molecular processes in whole cells and has the potential to be used in conjunction with electron microscopy and X-ray tomography to correlate structural and functional information. A SIM microscope for cryogenically preserved samples (cryoSIM) has recently been commissioned at the correlative cryo-imaging beamline B24 at the UK synchrotron. It was designed specifically for 3D imaging of biological samples at cryogenic temperatures in a manner compatible with subsequent imaging of the same samples by X-ray microscopy methods such as cryo-soft X-ray tomography. This video article provides detailed methods and protocols for successful imaging using the cryoSIM. In addition to instructions on the operation of the cryoSIM microscope, recommendations have been included regarding the choice of samples, fluorophores, and parameter settings. The protocol is demonstrated in U2OS cell samples whose mitochondria and tubulin have been fluorescently labelled.

Protocol for image registration of correlative soft X-ray tomography and super-resolution structured illumination microscopy images.

Correlation of 3D images acquired on different microscopes can be a daunting prospect even for experienced users. This protocol describes steps for registration of images from soft X-ray absorption contrast imaging and super-resolution fluorescence imaging of hydrated biological materials at cryogenic temperatures. Although it is developed for data generated at synchrotron beamlines that offer the above combination of microscopies, it is applicable to all analogous imaging systems where the same area of a sample is examined using successive non-destructive imaging techniques. For complete details on the use and execution of this protocol, please refer to Kounatidis et al. (2020).

Biofilm viability checker: An open-source tool for automated biofilm viability analysis from confocal microscopy images.

Quantifying biofilm formation on surfaces is challenging because traditional microbiological methods, such as total colony-forming units (CFUs), often rely on manual counting. These are laborious, resource intensive techniques, more susceptible to human error. Confocal laser scanning microscopy (CLSM) is a high-resolution technique that allows 3D visualisation of biofilm architecture. In combination with a live/dead stain, it can be used to quantify biofilm viability on both transparent and opaque surfaces. However, there is little consensus on the appropriate methodology to apply in confocal micrograph processing. In this study, we report the development of an image analysis approach to repeatably quantify biofilm viability and surface coverage. We also demonstrate its use for a range of bacterial species and translational applications. This protocol has been created with ease of use and accessibility in mind, to enable researchers who do not specialise in computational techniques to be confident in applying these methods to analyse biofilm micrographs. Furthermore, the simplicity of the method enables the user to adapt it for their bespoke needs. Validation experiments demonstrate the automated analysis is robust and accurate across a range of bacterial species and an improvement on traditional microbiological analysis. Furthermore, application to translational case studies show the automated method is a reliable measurement of biomass and cell viability. This approach will ensure image analysis is an accessible option for those in the microbiology and biomaterials field, improve current detection approaches and ultimately support the development of novel strategies for preventing biofilm formation by ensuring comparability across studies.

Access, barriers, and decisional regret in pursuit of fertility preservation among transgender and gender-diverse individuals.

OBJECTIVE: To query transgender and gender-diverse individuals on their desire for fertility preservation, perceived barriers to access care, and decisional regret. DESIGN: Cross-sectional. SETTING: Not applicable. PATIENT(S): A total of 397 gender-diverse individuals undergoing intake to the University of California Los Angeles Gender Health Program from January 2018 to March 2019. Seventy participated in a follow-up survey from September to October 2019 clarifying reproductive desires or intentions. INTERVENTION: Multiple-choice questionnaire. MAIN OUTCOME MEASURE(S): Perceived barriers to access fertility preservation and decisional regret surrounding choice to pursue fertility preservation as measured with the use of the validated Decision Regret Scale (scored 0 to 100). RESULT(S): Barriers to accessing care were primarily cost of treatment (36%), discontinuation/delay of hormonal therapy (19%), or worsening of gender dysphoria with treatment/pregnancy (11%). Respondents indicated that their family planning goals were addressed by primary care providers and/or medical endocrinologists (multiple responses allowed), but 37% stated that their family planning goals were not adequately addressed. Those who had made a firm decision to pursue or not pursue fertility treatment had mild decisional regret. Moderate-to-severe decisional regret was noted in those who were undecided regarding the pursuit of fertility perseveration before transition and in those who were interested in referral to reproductive endocrinology. CONCLUSION(S): Consultation with a reproductive endocrinologist may reduce decisional regret as well as clarify perceived barriers to fertility preservation in transgender and gender-diverse individuals interested in fertility preservation.

Imaging and Quantification of the Area of Fast-Moving Microbubbles Using a High-Speed Camera and Image Analysis.

An experimental and image analysis technique is presented for imaging cavitation bubbles and calculating their area. The high-speed imaging experimental technique and image analysis protocol presented here can also be applied for imaging microscopic bubbles in other fields of research; therefore, it has a wide range of applications. We apply this to image cavitation around dental ultrasonic scalers. It is important to image cavitation to characterize it and to understand how it can be exploited for various applications. Cavitation occurring around dental ultrasonic scalers can be used as a novel method of dental plaque removal, which would be more effective and cause less damage than current periodontal therapy techniques. We present a method for imaging the cavitation bubble clouds occurring around dental ultrasonic scaler tips using a high-speed camera and a zoom lens. We also calculate the area of cavitation using machine learning image analysis. Open source software is used for image analysis. The image analysis presented is easy to replicate, does not require programming experience, and can be modified easily to suit the application of the user.

High speed imaging of biofilm removal from a dental implant model using ultrasonic cavitation.

OBJECTIVES: Current instruments cannot clean in between dental implant threads and effectively remove biofilm from the rough implant surface without damaging it. Cavitation bubbles have the potential to disrupt biofilms. The aim of this study was to see how biofilms can be disrupted using non-contact cavitation from an ultrasonic scaler, imaged inside a restricted implant pocket model using high speed imaging. METHODS: Streptococcus sanguinis biofilm was grown for 7 days on dental implants. The implants were placed inside a custom made restricted pocket model and immersed inside a water tank. An ultrasonic scaler tip was placed 0.5mm away from the implant surface and operated at medium power or high power for 2s. The biofilm removal process was imaged using a high speed camera operating at 500 fps. Image analysis was used to calculate the amount of biofilm removed from the high speed images. Scanning electron microscopy was done to visualize the implant surface after cleaning. RESULTS: Cavitation was able to remove biofilm from dental implants. More biofilm was removed at high power. Scanning electron microscopy showed that the implant surface was clean at the points where the cavitation was most intense. High speed imaging showed biofilm removal underneath implant threads, in areas next to the ultrasonic scaler tip. SIGNIFICANCE: A high speed imaging protocol has been developed to visualize and quantify biofilm removal from dental implants in vitro. Cavitation bubbles from dental ultrasonic scalers are able to successfully disrupt biofilm in between implant threads.

Gynecologic and reproductive health in patients with pathogenic germline variants in DICER1.

OBJECTIVE: Germline pathogenic variation in DICER1 underlies a tumor-predisposition disorder with increased risk for cervical embryonal rhabdomyosarcoma and ovarian sex-cord stromal tumors, particularly Sertoli-Leydig cell tumors. The gynecologic and reproductive health of these females has not yet been described. METHODS: All female subjects recruited from November 2011 to July 2018 participating in an epidemiologic study of families with pathogenic DICER1 germline variation were included in this cross-sectional analysis. Participant evaluation included obstetric-gynecologic history, physical examination, hormone testing, pelvic ultrasound and record review. RESULTS: Of 64 females aged 2-72 years, fifteen underwent treatment for pleuropulmonary blastoma as children and three were treated for cervical embryonal rhabdomyosarcoma. Of nine patients reporting a history of ovarian tumors, all presented with virilization or amenorrhea; eight occurred in adolescence. Post-pubertal females with no history of ovarian tumors experienced normal pubertal development, reported regular menstrual cycles, were fertile and underwent natural menopause at median age of 52 years. Thirty-two of 33 women who tried to conceive successfully delivered liveborn children. Of these 32, 10 experienced pregnancy-related thyroid enlargement resulting in thyroidectomy within one year of pregnancy; nine others had undergone pre-pregnancy thyroidectomy. CONCLUSION: In these DICER1-carrier females, DICER1-related gynecological tumors occurred during childhood or adolescence in some after which women generally experienced healthy reproductive lives. Individual education and screening for these tumors is warranted. The high rate of DICER1-related multinodular goiter resulting in pre- and post-pregnancy thyroidectomy underscores the importance of thyroid monitoring during pregnancy to ensure maternal and fetal wellbeing.

Which Parameters Affect Biofilm Removal with Acoustic Cavitation? A Review.

Bacterial biofilms are a cause of contamination in a wide range of medical and biological areas. Ultrasound is a mechanical energy that can remove these biofilms using cavitation and acoustic streaming, which generate shear forces to disrupt biofilm from a surface. The aim of this narrative review is to investigate the literature on the mechanical removal of biofilm using acoustic cavitation to identify the different operating parameters affecting its removal using this method. The properties of the liquid and the properties of the ultrasound have a large impact on the type of cavitation generated. These include gas content, temperature, surface tension, frequency of ultrasound and acoustic pressure. For many of these parameters, more research is required to understand their mechanisms in the area of ultrasonic biofilm removal, and further research will help to optimise this method for effective removal of biofilms from different surfaces.

Maternal and neonatal outcomes associated with infertility.

Objective: To investigate perinatal outcomes in a cohort of fertile and infertile nulliparous women. Design: Retrospective cohort study. Setting: Academic medical center. Patients: All nulliparous women delivering singletons ≥24-week gestation at our center from 1 January 2012 to 31 December 2012 were included. Women were classified into two groups - fertile and infertile - based on a chart review at the time of delivery. Outcome measure: Perinatal outcomes of interest included mode of delivery, gestational age at delivery, and birth weight. Results: A total of 3293 mother/infant dyads fulfilled the inclusion criteria. Of these, 277 women (8.4%) were classified as infertile. Infertile women were significantly older than fertile women. In bivariate analyses, infertile women were more likely to undergo cesarean delivery (51.8 versus 36.1%, p < .001) and deliver at an earlier gestational age (38.9 ± 2.3 versus 39.4 ± 1.7 weeks, p < .0001). Infertility was no longer significantly associated with cesarean delivery after adjusting for maternal age. Infertility remained associated with an earlier gestational age at delivery after adjusting for maternal age and maternal race (ß coefficient -0.42, 95%CI -0.64, -0.2). There was no difference in infant birth weight. Late preterm deliveries (34-36 completed gestational weeks) accounted for 8.3% of deliveries for infertile women compared to 4.3% for fertile women (p = .032). Conclusions: We conclude that the increased risk of cesarean section associated with infertility is driven by maternal age. Late preterm infants represent a key cohort for further evaluation in the perinatal outcomes of infertile women.

Effect of Zr Addition on the Corrosion of Ti in Acidic and Reactive Oxygen Species (ROS)-Containing Environments.

The effect of systematic Zr additions on the corrosion behavior of Ti was studied in both acidic and reactive oxygen species (ROS) containing environments, including macrophage cell culture, simulating inflammation associated with metallic implants. Electrochemical measurements on commercially pure (CP) Ti, Zr, and TiZr alloys showed that increasing Zr additions progressively enhanced Ti passivity in both acidic (HCl) and oxidative (H2O2) environments. However, a Ti50Zr alloy was found with increased pitting susceptibility. Corrosion was also evaluated using mass-spectrometry to determine metal ion release following exposure of the alloys to THP-1 macrophage cell cultures, transformed into either their M1 (inflammatory states) or M2a (tissue repair states) phenotypes. The magnitude of ion release was reduced with increasing Zr contents, consistent with electrochemical observations. Nevertheless, optimized Zr content in Ti should balance both passivity and pitting resistance.

Impact of fertility treatment on severe maternal morbidity.

OBJECTIVE: To determine if fertility treatment is associated with increased risk of severe maternal morbidity (SMM) compared with spontaneous pregnancies. DESIGN: Retrospective cohort study. SETTING: Academic medical center. PATIENT(S): In 2012, 6,543 women delivered live births >20 weeks gestation at our center. Women were categorized based on mode of conception: in vitro fertilization (IVF), non-IVF fertility treatment (NIFT), or spontaneous pregnancies. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The main outcome was presence of true SMM, such as eclampsia, respiratory failure, and peripartum hysterectomy. Deliveries were screened with the use of: 1) International Classification of Diseases 9 codes; 2) prolonged postpartum stay; 3) maternal intensive care unit admissions, and 4) blood transfusion. The charts of women meeting the screening criteria were reviewed to identify true SMM based on a previously validated method, recognizing that medical record review is the criterion standard. RESULT(S): Of the 6,543 deliveries, 246 (3.8%) were IVF conceptions and 109 (1.7%) NIFT conceptions. Sixty-nine cases of true SMM were identified (1.1%). In multivariate analyses, any fertility treatment (IVF + NIFT) was associated with increased risk of SMM compared with spontaneous conceptions. In a subset analysis of singletons only, the association between any fertility treatment (IVF + NIFT) and SMM was not statistically significant. CONCLUSION(S): Overall, fertility treatment increased risk for SMM events. Given the limited sample size, the negative finding with singleton gestations is inconclusive. Larger multicenter studies with accurate documentation of fertility treatment and SMM cases are needed to further clarify the risk associated with singletons.

High Speed Imaging of Cavitation around Dental Ultrasonic Scaler Tips.

Cavitation occurs around dental ultrasonic scalers, which are used clinically for removing dental biofilm and calculus. However it is not known if this contributes to the cleaning process. Characterisation of the cavitation around ultrasonic scalers will assist in assessing its contribution and in developing new clinical devices for removing biofilm with cavitation. The aim is to use high speed camera imaging to quantify cavitation patterns around an ultrasonic scaler. A Satelec ultrasonic scaler operating at 29 kHz with three different shaped tips has been studied at medium and high operating power using high speed imaging at 15,000, 90,000 and 250,000 frames per second. The tip displacement has been recorded using scanning laser vibrometry. Cavitation occurs at the free end of the tip and increases with power while the area and width of the cavitation cloud varies for different shaped tips. The cavitation starts at the antinodes, with little or no cavitation at the node. High speed image sequences combined with scanning laser vibrometry show individual microbubbles imploding and bubble clouds lifting and moving away from the ultrasonic scaler tip, with larger tip displacement causing more cavitation.

Increased medial tibial slope in teenage pediatric population with open physes and anterior cruciate ligament injuries.

PURPOSE: Variations in bony morphology have been associated with anterior cruciate ligament (ACL) injury risk. The primary aim of this study was to compare the tibial slope in the teenage pediatric population with open physes, with and without ACL injury. The secondary aims were to compare the notch width index (NWI) and determine the influence of gender and age on tibial slope and NWI. METHODS: Thirty-nine teenage pediatric subjects were included in this study, 16 with and 23 without ACL injury. Medial tibial slope and lateral tibial slope and NWI as measured on plain radiographs were compared between the groups using an independent t-test. Comparison of tibial slope and NWI was also performed between male and female subjects. Pearson correlation coefficient between age and tibial slope and NWI was calculated. RESULTS: The medial tibial slope averaged 10.2 (±3.9) degrees, the lateral tibial slope 11.5 (±3.9) degrees, and the NWI 0.26. There was a significant difference in medial tibial slope between the ACL-injured (12.1 degrees) subjects and the controls (8.9 degrees) (P = 0.009). There was no significant difference in lateral tibial slope or NWI between the groups. There was no significant difference in the medial tibial slope and lateral tibial slope and NWI between the male and female subjects. Subject age was not correlated with medial tibial slope and lateral tibial slope or NWI. CONCLUSION: There was an increased medial tibial slope in ACL-injured teenagers with open physes, when compared to a control group of teenager with open physes without ACL injury.