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The authors assemble and analyse previously generated mycobiome data linked to geographical locations across the world. They describe the distribution of fungal taxa and show that climate is an important driver of fungal biogeography and that fungal diversity appears to be concentrated at high latitudes.

Purpose: To develop and apply a neural network for quantification of endothelial corneal graft detachment using anterior segment (AS) OCT. Design: Training and validation of a neural network and application within a prospective cohort. Participants: Patients two weeks after Descemet membrane endothelial keratoplasty. Methods: Investigators manually labeled the posterior cornea and the graft in cross-sectional images of rotational AS OCT scans. Neural networks for image segmentation were trained to identify the area of graft detachment on cross-sectional images. The best-performing neural network with the lowest misclassification (Youden index) and highest spatial overlap with the ground truth (Dice coefficient) was selected and evaluated in a separate dataset. Three-dimensional maps of the area and volume of graft detachment were calculated. For application, the neural network’s rating on the detachment was compared with slit-lamp–based ratings of cornea specialists on the same day as the AS OCT imaging took place. Main Outcome Measures: Youden index and Dice coefficient. Results: Neural networks were trained on 27 AS OCT scans with 6912 labeled images. Among 48 combinations of probability thresholds and epoch states, the best-performing neural network showed a Youden index of 0.99 and a Dice coefficient of 0.77, indicating low misclassification and good spatial overlap on individual image segmentation. In the validation set unknown to the neural network with 20 scans (5120 images), the Youden index was 0.85 and the Dice coefficient was 0.73, and a high overall performance compared with the manually labeled ground truth (R2 = 0.90). In the application set with 107 eyes, the neural network estimated the mean percent detachment larger than the cornea specialist (mean difference, 8.2 percentage points; 95% confidence interval, 6.2–10.2). Masked review of 42 AS OCTs with more than ±10 percentage points difference in ratings showed that clinicians underestimated the true detachment in cases with significant detachment requiring intervention. Conclusions: Deep learning-based segmentation of AS OCT images quantified the percent and the volume of DMEK graft detachment with high precision. Fully automated 3-dimensional quantification of graft detachment is highly sensitive, particularly in corneas with a significant amount of detachment, and may support decision making.

Manual material handling (MMH) is considered as one of the main contributors to low back pain. While males traditionally perform MMH tasks, recently the number of females who undertake these physically-demanding activities is also increasing. To evaluate the risk of mechanical injuries, the majority of previous studies have estimated spinal forces using different modeling approaches that mostly focus on male individuals. Notable sex-dependent differences have, however, been reported in torso muscle strength and anatomy, segmental mass distribution, as well as lifting strategy during MMH. Therefore, this study aimed to use sex-specific models to estimate lumbar spinal and muscle forces during static MHH tasks in 10 healthy males and 10 females. Motion-capture, surface electromyographic from select trunk muscles, and ground reaction force data were simultaneously collected while subjects performed twelve symmetric and asymmetric static lifting (10 kg) tasks. AnyBody Modeling System was used to develop base-models (subject-specific segmental length, muscle architecture, and kinematics data) for both sexes. For females, female-specific models were also developed by taking into account for the female’s muscle physiological cross-sectional areas, segmental mass distributions, and body fat percentage. Males showed higher absolute L5-S1 compressive and shear loads as compared to both female base-models (25.3% compressive and 14% shear) and female-specific models (41% compressive and 23.6% shear). When the predicted spine loads were normalized to subjects’ body weight, however, female base-models showed larger loads (9% compressive and 16.2% shear on average), and female-specific models showed 2.4% smaller and 9.4% larger loads than males. Females showed larger forces in oblique abdominal muscles during both symmetric and asymmetric lifting tasks, while males had larger back extensor muscle forces during symmetric lifting tasks. A stronger correlation between measured and predicted muscle activities was found in females than males. Results indicate that female-specific characteristics affect the predicted spinal loads and must be considered in musculoskeletal models. Neglecting sex-specific parameters in these models could lead to the overestimation of spinal loads in females.

LiDAR-based segmentation of urban tree canopies and their physical properties (canopy height, canopy diameter, 3D surface and volume) is a replicable, complementary and useful information source for urban ecosystem condition accounts, and an important basis for ecosystem service modeling and valuation. However, using available LiDAR data collected for municipal purposes other than vegetation mapping (such as for example engineering) entails a level of accuracy which may limit the usefulness of the data for change detection in ecosystem accounts. To account for changes in the urban tree canopy of Oslo (capital city of Norway) between 2011 and 2017, a segmentation model was developed based on available airborne LiDAR data scanned for general purposes. The results from the entire built-up area of Oslo indicate a general increase in the number of tall trees (>15 m) and a moderate increase in the number of small trees (10 m). The total tree canopy area within the Small House plan area decreased by 1.04%, with a corresponding 2.13% decrease in the tree canopy volume. With respect to the segmentation accuracy, the changes in aggregate tree canopy cover are too small to determine canopy change with confidence. This study demonstrates the potential for identifying ecosystem condition indicators as well as the limitations of using general purpose LiDAR data to improve the precision of urban ecosystem accounting. For future ecosystem service accounting in urban environments, we recommend that municipalities implement data acquisition programs that combine concurrent field data sampling and LiDAR campaigns designed for urban tree canopy detection, as part of general urban structural inventorying. We recommend using LiDAR and satellite remote sensing data depending on canopy densities. We also recommend that future tree canopy segmentation is done within a cloud-computing environment to ensure sufficient geoprocessing capacity.

Raw coffee beans are seen as valuable sources of bioactive compounds, such as alkaloids and chlorogenic acids. In this study, an efficient and eco-friendly method for the simultaneous extraction of caffeine, trigonelline, and chlorogenic acids from raw beans of two coffee species was developed, using green solvents and an unconventional method of extraction (assisted by ultrafast rotary disintegrator/homogenizer (UT-AE)). The experimental extraction conditions were optimized according to a completely randomized design (CRD), considering the following variables: solvent type (four deep eutectic solvents (DESs), water, and aqueous choline chloride solution (50 wt.%); temperature (25, 45, and 65 °C); and extraction technique (solid-liquid extraction with agitated heating and assisted by ultrafast rotary disintegrator/homogenizer). The extract obtained with choline chloride solution (50 wt.%) exhibited high total phenolic compounds and the highest antioxidant capacity. An analysis using high-performance liquid chromatography with a diode array detector (HPLC-DAD) indicated the presence of chlorogenic acids, caffeine, and trigonelline in all the extracts, in different amounts. The results obtained by the analysis of phenolic compounds and HPLC indicated that the aqueous solution of choline chloride (50% wt.%) was the most suitable solvent for the extraction of chlorogenic acids, while the water-based extracts showed high values of caffeine and trigonelline. DESs, in turn, seems to promote a protective effect on the antioxidant activity of biomolecules.

The energy sector is one of the most important pollutants in the atmosphere and causes significant emissions of greenhouse gases. In Russia, coal is the main contributor to the fossil fuel consumption of thermal power plants and boilers, thus affecting atmospheric air pollution by releasing particulate matter and nitrogen oxides, which are strongly associated with a negative impact on human health. This problem is especially acute for the resource regions of Yenisei, Siberia, a 2.5+ mln sq km macro-region in the very heart of Russia. In this paper, we analyze the impact of the structure of electricity and heat generation on emissions of pollutants and climate-active gases in Yenisei, Siberia, and give an overview of their social, ecological, and economic effects. More than 75% of electricity in Yenisei, Siberia, is produced by hydroelectric power plants that do not pollute the atmosphere. The rest of the electricity is generated in the cogeneration mode by thermal power plants, which are cores of the heat supply designs of cities. The share of individual coal-powered heat sources is still high. A detailed analysis of existing equipment and technologies at existing thermal power plants is needed to select options for their modernization to reduce emissions while keeping coal in use. Our calculations for the biggest cities of Krasnoyarsk Krai show that investments in the transition to heating with pellets will require RUB 184.7 million for Nazarovo and RUB 313.9 million for Kansk. At the same time, switching to electric heating is more than twice as expensive: RUB 498.6 million for Nazarovo and RUB 847.5 million for Kansk. The additional costs will range from RUB 21 to RUB 45.4 thousand per household per year for the pellet variant and from RUB 56.8 to RUB 122.5 thousand per year for electric heating, which could triple the annual heating costs. Thus, these options are unlikely to be implemented without direct state support. We argue that creating an attractive living environment in Yenisei, Siberia, must begin with intensive public investment in mitigating the environmental externalities caused by coal emissions.

In patients with relapsed/refractory Burkitt lymphoma (r/r BL), overall survival (OS) is poor, and effective therapies and evidence for the best therapy are lacking. The monoclonal antibody blinatumomab may represent a novel option. However, only limited data on the use of blinatumomab in r/r BL are so far available. This multi-center, retrospective case series investigated nine patients with r/r BL treated with blinatumomab. The safety of blinatumomab was assessed with respect to frequency and severity of adverse effects (AEs) infections, cytokine release syndrome (CRS) and neurotoxicity. Progression-free survival (PFS), OS and overall response rate (ORR) were analyzed to assess efficacy. No AEs > grade 2 occurred, and AEs were generally treatable and fully reversible. The best response to blinatumomab was complete remission in 3/9 patients and partial remission in 2/9, whilst 4/9 presented with progressive disease. Median PFS and OS were 2 and 6 months, respectively, ranging from 5 days to 32 months and 11 days to 32 months, respectively. Blinatumomab treatment was a successful bridging treatment to stem cell transplantation in 3/9 patients. The response to blinatumomab varied widely, and only one patient survived longer term, but activity in patients with r/r BL was evident in some patients, with its use being safe, warranting its prospective investigation.

A meta-analytic review of 33 studies and 41 independent samples was conducted of the effectiveness of community-based mental health and behavioral programs for low-income urban youth. Findings indicated positive effects, with an overall mean effect of 0.25 at post-test. While this is comparable to previous meta-analytic intervention research with more diverse samples, it stands in contract to findings of the school-based review (0.08 at post-test) focused on this population. The current review found type of intervention to significantly moderate effects, with effects highest for programs that were environmentally based. In fact, effect sizes for programs that did not target the environment were not significant. Findings are discussed with an emphasis on environmental influences, including the differing contexts affecting school versus community-based interventions conducted with low income, urban youth. [This article was published in "Clinical Psychology: Science and Practice," v19 n2 p195-215 Jun 2012.]

We have sequenced the genomes of 110 small cell lung cancers (SCLC), one of the deadliest human cancers. In nearly all the tumours analysed we found bi-allelic inactivation of TP53 and RB1, sometimes by complex genomic rearrangements. Two tumours with wild-type RB1 had evidence of chromothripsis leading to overexpression of cyclin D1 (encoded by the CCND1 gene), revealing an alternative mechanism of Rb1 deregulation. Thus, loss of the tumour suppressors TP53 and RB1 is obligatory in SCLC. We discovered somatic genomic rearrangements of TP73 that create an oncogenic version of this gene, TP73Δex2/3. In rare cases, SCLC tumours exhibited kinase gene mutations, providing a possible therapeutic opportunity for individual patients. Finally, we observed inactivating mutations in NOTCH family genes in 25% of human SCLC. Accordingly, activation of Notch signalling in a pre-clinical SCLC mouse model strikingly reduced the number of tumours and extended the survival of the mutant mice. Furthermore, neuroendocrine gene expression was abrogated by Notch activity in SCLC cells. This first comprehensive study of somatic genome alterations in SCLC uncovers several key biological processes and identifies candidate therapeutic targets in this highly lethal form of cancer.

Global warming and climate change have become hot topics that incite debate, inspire scientific research, and influence international policy. However, the scientific research that provides the past climate and environmental information upon which contemporary environmental change is measured, receives little attention in high school curriculum. Paleoecology, the study of ancient ecosystems, provides a unifying theme for teaching multiple high school science curriculum concepts involving global environmental change. As a teaching tool, paleoecology establishes a framework linking concepts such as geologic time, climate change, adaptation, survival, extinction, human impact and ecological interactions that are often taught separately. This article provides a brief overview of how the science of paleoecology can be introduced to students and incorporated into the curriculum through simple activities. The activities outlined here include using elevation maps or Gazetteers to investigate potential sites where marine fossils may be found far from the ocean, using multiple biological proxies to measure climate change, and creating models to demonstrate the impact of sea level rise on coastal ecosystems. These activities provide numerous opportunities for the students to discuss the scientific research associated with climate change, the economic impacts of changing climate, and how science may influence policy regarding climate change mitigation. (Contains 1 figure and 1 table.)

Background: De novo steatosis is the main criteria for non-alcoholic fatty liver disease (NAFLD), which is becoming a clinically relevant comorbidity in HIV-infected patients. This may be due to the HIV virus itself, as well as long-term toxicities deriving from antiretroviral therapy. Therefore, HIV infected patients require prevention and monitoring regarding NAFLD. Methods: This study investigated the differential role of body mass index (BMI) and combination antiretroviral treatment (cART) drugs on NAFLD progression. This single center prospective longitudinal observational study enrolled HIV monoinfected individuals between August 2013 to December 2018 with yearly visits. Each visit included liver stiffness and steatosis [defined as controlled attenuation parameter (CAP)>237 dB/m] assessment by annually transient elastography using an M- or XL-probe of FibroScan, and calculation of the novel FibroScan-AST (FAST) score. Risk factors for denovo/progressed steatosis and tripling of FAST-score increase were investigated using Cox regression model with time-dependent covariates. Findings: 319 monoinfected HIV positive patients with at least two visits were included into the study, of which 301 patients had at least two valid CAP measurements. 51·5%(155) patients did not have steatosis at first assessment, of which 45%(69) developed steatosis during follow-up. A BMI>23 kg/m2 (OR: 4·238, 95% CI: 2·078–8·938; p < 0·0001), tenofovir-alafenamid (TAF) (OR: 5·073, 95% CI: 2·362–10·899); p < 0·0001) and integrase strand transfer inhibitors (INSTI) (OR: 2·354, 95% CI: 1·370–4·048; p = 0·002), as well as type 2 diabetes mellitus (OR: 7·605, 95% CI: 2·315–24·981; p < 0·0001) were independent predictors of de novo steatosis in multivariable analysis. Tenofovir disoproxilfumarate (TDF) was associated with a lower risk for weight gain and steatosis progression/onset using CAP value (HR: 0·28, 95% CI: 0·12–0·64; p = 0·003) and FAST scores (HR: 0·31, 95% CI: 0·101–0·945; p = 0·04). Interpretation: Steatosis can develop despite non-obese BMI in patients with HIV monoinfection under cART, especially in male patients with BMI over 23 kg/m2. While TAF and INSTI increase the risk of progression of steatosis, TDF was found to be independently associated with a lower risk of a clinically significant weight gain and thereby, might slow down development and progression of steatosis. Funding: There was no additional funding received for this project. All funders mentioned in the ‘declaration of interests’ section had no influence on study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Background and study aims Colonoscopy is considered the gold standard for decreasing colorectal cancer incidence and mortality. Optical diagnosis of colorectal polyps (CRPs) is an ongoing challenge in clinical colonoscopy and its accuracy among endoscopists varies widely. Computer-aided diagnosis (CAD) for CRP characterization may help to improve this accuracy. In this study, we investigated the diagnostic accuracy of a novel algorithm for polyp malignancy classification by exploiting the complementary information revealed by three specific modalities. Methods We developed a CAD algorithm for CRP characterization based on high-definition, non-magnified white light (HDWL), Blue light imaging (BLI) and linked color imaging (LCI) still images from routine exams. All CRPs were collected prospectively and classified into benign or premalignant using histopathology as gold standard. Images and data were used to train the CAD algorithm using triplet network architecture. Our training dataset was validated using a threefold cross validation. Results In total 609 colonoscopy images of 203 CRPs of 154 consecutive patients were collected. A total of 174 CRPs were found to be premalignant and 29 were benign. Combining the triplet network features with all three image enhancement modalities resulted in an accuracy of 90.6 %, 89.7 % sensitivity, 96.6 % specificity, a positive predictive value of 99.4 %, and a negative predictive value of 60.9 % for CRP malignancy classification. The classification time for our CAD algorithm was approximately 90 ms per image. Conclusions Our novel approach and algorithm for CRP classification differentiates accurately between benign and premalignant polyps in non-magnified endoscopic images. This is the first algorithm combining three optical modalities (HDWL/BLI/LCI) exploiting the triplet network approach.

ABSTRACT Current procurement of specimens for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection requires trained personnel and dedicated equipment. We compared standard nasopharyngeal swabs with self-collected gargle lavage fluid obtained from 80 mostly symptomatic outpatients. After RNA extraction, RT-PCR to detect SARS-CoV-2 was performed. Qualitative results obtained with the paired samples from individual outpatients were 100% congruent. Therefore, self-collected gargle lavage fluid can serve as a suitable specimen for coronavirus disease 2019 (COVID-19) testing in outpatients. IMPORTANCE The SARS-CoV-2 pandemic still strains health care systems worldwide. While COVID-19 testing is considered an essential pillar in combating this infectious disease, shortages in supplies and trained health care personnel often limit the procurement of patient samples, in particular in outpatient settings. Here, we compared the simple self-collection of gargle lavage fluid with the gold standard nasopharyngeal swab as a specimen for COVID-19 testing. By finding complete congruence of results obtained with paired samples of a sizeable patient cohort, our results strongly support the idea that the painless self-collection of gargle lavage fluid provides a suitable and uncomplicated sample for reliable SARS-CoV-2 detection.

BackgroundAdequate bowel preparation is essential for the visualization of the colonic mucosa during colonoscopy. However, the rate of inadequate bowel preparation is still high, ranging from 18% to 35%; this may lead to a higher risk of missing clinically relevant lesions, procedural difficulties, prolonged procedural time, an increased number of interval colorectal carcinomas, and additional health care costs. ObjectiveThe aims of this study are to compare bowel preparation instructions provided via a personalized smartphone app (Prepit, Ferring B V) with regular written instructions for bowel preparation to improve bowel preparation quality and to evaluate patient satisfaction with the bowel preparation procedure. MethodsEligible patients scheduled for an outpatient colonoscopy were randomized to a smartphone app group or a control group. Both the groups received identical face-to-face education from a research physician, including instructions about the colonoscopy procedure, diet restrictions, and laxative intake. In addition, the control group received written information, whereas the smartphone app group was instructed to use the smartphone app instead of the written information for the actual steps of the bowel preparation schedule. All patients used bisacodyl and sodium picosulfate with magnesium citrate as laxatives. The quality of bowel preparation was scored using the Boston Bowel Preparation Scale (BBPS) by blinded endoscopists. Patient satisfaction was measured using the Patient Satisfaction Questionnaire-18. ResultsA total of 87 patients were included in the smartphone app group and 86 in the control group. The mean total BBPS score was significantly higher in the smartphone app group (mean 8.3, SD 0.9) than in the control group (mean 7.9, SD 1.2; P=.03). The right colon showed a significantly higher bowel preparation score in the smartphone app group (mean 2.7, SD 0.5 vs mean 2.5, SD 0.6; P=.04). No significant differences were observed in segment scores for the mean transverse colon (mean 2.8, SD 0.4 vs mean 2.8, SD 0.4; P=.34) and left colon (mean 2.8, SD 0.4 vs mean 2.6, SD 0.5; P=.07). General patient satisfaction was high for the smartphone app group (mean 4.4, SD 0.7) but showed no significant difference when compared with the control group (mean 4.3, SD 0.8; P=.32). ConclusionsOur personalized smartphone app significantly improved bowel preparation quality compared with regular written instructions for bowel preparation. In particular, in the right colon, the BBPS score improved, which is of clinical relevance because the right colon is considered more difficult to clean and the polyp detection rate in the right colon improves with improvement of bowel cleansing of the right colon. No further improvement in patient satisfaction was observed compared with patients receiving regular written instructions. Trial RegistrationClinicalTrials.gov NCT03677050; https://clinicaltrials.gov/ct2/show/NCT03677050

This article describes how Extension professionals and university faculty collaborated on the planning, implementation, and evaluation of a conference to address a key societal issue. Needs assessment and evaluation results are presented as well as an explanation of how results were used in planning future conferences. With limited Extension budgets and personnel, partnering among Extension specialists, university faculty, and experts in relevant topics is a valuable and efficient way to provide the most impact for a community. Developing multiyear conferences to address significant societal issues may be a new way to increase Extension's impact.

Research on sex-related differences in Autism Spectrum Disorder (ASD) has been impeded by small samples. We pooled 28 datasets from 18 sites across nine European countries to examine sex differences in the ASD phenotype on the ADI-R (376 females, 1763 males) and ADOS (233 females, 1187 males). On the ADI-R, early childhood restricted and repetitive behaviours were lower in females than males, alongside comparable levels of social interaction and communication difficulties in females and males. Current ADI-R and ADOS scores showed no sex differences for ASD severity. There were lower socio-communicative symptoms in older compared to younger individuals. This large European ASD sample adds to the literature on sex and age variations of ASD symptomatology.

Sleep fosters the generation of explicit knowledge. Whether sleep also benefits implicit intuitive decisions about underlying patterns is unclear. We examined sleep's role in explicit and intuitive semantic coherence judgments. Participants encoded sets of three words and after a sleep or wake period were required to judge the potential convergence of these words on a common fourth associate. Compared with wakefulness, sleep increased the number of explicitly named common associates and decreased the number of intuitive judgments. This suggests that sleep enhances the extraction of explicit knowledge at the expense of the ability to make intuitive decisions about semantic coherence.

Collaboration between teachers constitutes an important predictor for the successful implementation of digital media in schools and teaching. The present contribution examines the supporting conditions of ICT (information and communications technology)-related teacher collaboration as a feature of school quality in six selected European educational systems on the basis of the instruments and data administered by the International Association for the Evaluation of Educational Achievement's study of ICILS 2013 (International Computer and Information Literacy Study 2013). Along the ICILS 2013 theoretical framework, predictors on the level of the school and classroom, antecedents regarding teachers' attitudes and competencies, variables concerning processes at school and in class, as well as teachers' background variables, are analysed. The regression analysis shows no consistent differences regarding the coefficients of different predictors with respect to ICT-related collaboration in the countries where teachers have expressed a high, medium or low rate of agreement with ICT-related collaboration.

Atypical gaze processing has been reported in children with autism spectrum disorders (ASD). Here we explored how infants at risk for ASD respond behaviorally to others' direct gaze. We assessed 10-month-olds with a sibling with ASD (high risk group; n = 61) and a control group (n = 18) during interaction with an adult. Eye-tracking revealed less looking at the adult in the high risk group during 300-1000 ms after the adult initiated direct gaze: a short alteration that is likely to go unnoticed by the naked eye. Data aggregated over longer segments (the traditional eye-tracking approach) showed no group differences. Although findings are limited by lack of outcome data, they are in line with theories linking atypical eye processing to the emergence of ASD.

Background: Inter-operator variation in morphology-based embryo assessment is known to be an ongoing issue in in vitro fertilisation (IVF). This includes inconsistent annotation of embryo morphology and disagreement in the predictive weightings amongst different morphological features. Although currently underexplored, subjectivity reduction using statistical tools is considered clinically valuable, and is more affordable than newer approaches such as time-lapse imaging. Aim: This study aims to develop a robust numerical D5 embryo ranking system by incorporating several patient and embryo characteristics arising from routine static D5 assessment. Method: A total of 8866 autologous-oocyte single fresh D5 transfers performed at 14 Monash IVF clinics during 2012-2018 were retrospectively analyzed, with repeat cycles from the same patients excluded to avoid clustering effects. The dataset was randomly split into two subsets at 60:40 ratio, including one (n=5274) used for regression analysis and model development and the other (n=3592) for validation. Multiple logistic regression was performed to evaluate live birth prediction by several variables, expressed by odds ratio (OR) and 95% confidence interval (CI). A numerical ranking system was developed based on calculated weightings of contributing factors and subsequently tested using receiver operating characteristics (ROC) as expressed by area under the ROC curve (AUC). Results: All included variables were significantly predictive of live birth, including maternal age (OR=1.465, 1.364-1.574, P=0.000), assessment timing (OR=1.096, 1.020-1.177, P=0.012), embryo developmental stage (OR=1.341, 1.244-1.445, P=0.000) and morphological sore (OR=1.520, 1.392-1.661, P=0.000). Increments of resultant numerical ranking scores ([Formula: see text]3, 3-3.99, 4-4.99, and [Formula: see text]5) were associated with increasing live birth rates (9.4%, 27.2%, 43.6%, and 54.9%; respectively). ROC analysis indicated significantly predictive power of the numerical ranking system in both the development (AUC=0.690, 0.675-0.704, P=0.000) and validation (AUC=0.685, 0.667-0.703, P=0.000) subsets. Conclusion: A robust numerical model can be developed for D5 embryo selection, potentially leading to improved inter-operator subjectivity for embryo assessment.

Background: There is currently no consensus regarding developmental cutoff for fresh transfer of slow developing day 5 embryos on. Literature is sparse regarding true prognosis of such embryos. Aim: To investigate live birth and birthweight outcomes of slow developing day 5 transferred embryos. Method: This retrospective multi-center study included 1213 consecutive autologous-oocyte single fresh day 5 transfers performed at 4 Monash IVF clinics during 2016-2019. Repeat cycles by same patients were excluded to avoid clustering effects. Live birth and birthweight were followed up in all pregnancies. Multiple regression was performed to investigate associations between slow day 5 development (defined as ≤ early blastocyst) and (a) live birth, (b) birthweight, and (c) gestation-adjusted birthweight (Z score) to account for gestational age and gender. Results were expressed as adjusted odds ratio (aOR) with 95% confidence interval (CI). Results: No implantation was achieved when day 5 embryo failed to reach early blastocyst stage (i.e. ≤morula, n=76). Live birth rate was significantly lower following transfer of early blastocysts (n=237, 16%), in comparison to expanding (n=329, 27%, P=0.001), expanded (n=392, 41%, P=0.000), and ≥hatching blastocysts (n=169, 44%, P=0.000). After adjusting for maternal age, hours post insemination at day 5 assessment, number of oocytes collected, number of 2PNs, and number of embryos frozen; multiple logistic regression showed a significantly reduced likelihood of live birth resulting from early blastocysts in reference to expanding (aOR=0.584, 0.371-0.917, P=0.020), expanded (aOR=0.322, 0.208-0.501, P=0.000), or ≥hatching stages (aOR=0.255, 0.147-0.443, P=0.000). However, early blastocysts resulting in live birth (n=39) did not alter birthweight or Z score in reference to expanding (n=90, P>0.05 respectively), expanded (n=160, P>0.05 respectively), or ≥hatching (n=75, P>0.05 respectively). Conclusion: There is no clinical value of fresh transferring day 5 embryos at ≤morula stage. Fresh early blastocysts lead to a reduced live birth rate but not birthweight outcomes.

Background: Non-obstructive azoospermia (NOA) is a severe form of male factor infertility, evident in approximately 5% of infertile couples seeking fertility treatment. The combination of microsurgical testicular sperm extraction (mTESE) and ICSI has become one of the treatment options, with sperm motility positively associated with outcome success rates. Few studies have investigated whether use of fresh or thawed gamete combinations affect clinical outcomes. Aim: To determine the clinical outcomes of ICSI cycles using mTESE recovered testicular sperm of NOA patients and whether these are affected by the use of fresh or thawed gametes. Method: A retrospective study of NOA patients who underwent mTESE between March 2017 and September 2020. Clinical outcomes investigated included successful surgical sperm retrieval (SSR), sperm motility, fertilisation, embryo utilisation and live-birth/ongoing pregnancy rates. Results: A total of 105 NOA patients underwent mTESE, with a 65.7% successful SSR. In total 58 patients contributed to 72 ICSI cycles, with a 37.5% 2PN fertilisation rate and 29.4% live-birth/ongoing pregnancy rate. Cycles using only motile sperm had a greater fertilisation rate than those using a motile/non-motile sperm combination (51.7% vs 31.1%, p[Formula: see text]0.05). Compared with fresh, thawed testicular spermatozoa didn’t affect clinical outcomes, including live-birth/ongoing pregnancy rates (Fresh 18.2%; Thawed 17.9%; Odds ratio (OR) 1.02 (0.34-2.86) p=0.98). In contrast, use of thawed oocytes decreased fertilisation rate [Fresh-oocytes: 45.0%; Vitrified-oocytes: 35.5%; OR 1.47(1.03-2.10), p=0.03] but had no differences for other clinical outcomes, including live-birth/ongoing pregnancy rate (p=0.74). Importantly, injection of exclusively non-motile-spermatozoa (n=26 cycles) resulted in a very low fertilisation rate (2.7%) and no live-birth/ongoing pregnancies were recorded. Conclusion: Microdissection TESE is an effective treatment for NOA patients, with no clear negative effects of using thawed spermatozoa, however, the use of vitrified oocytes requires further investigation. The use of non-motile spermatozoa in mTESE ICSI cycles is discouraged.

Background: Intracytoplasmic sperm injection (ICSI) usage ([Formula: see text]64% of autologous cycles) has increased in Australia as an attempt to avoid low fertilisation and total fertilisation failure after standard in vitro fertilization (IVF). ICSI and to a lesser extent IVF, bypass key sperm maturation (hyperactivation, capacitation/acrosome reaction) events that naturally occur in the female tract and are vital for successful fertilisation. Current commercial sperm preparation media are not fully designed to induce these changes in sperm prior to insemination. Aim: To improve fertilisation rates following IVF by increasing sperm capacitation and hyperactivation between sperm preparation and insemination ultilising a new sperm medium (SpermFAST). Method: Sperm from 12 consenting normospermic men were incubated in either G-IVF+ (Vitrolife) or SpermFAST (UoA/Monash IVF) following a direct swim-up. Measures of capacitation, hyperactivation, sperm binding, acrosome reaction and oxidative DNA damage were assessed. Further, sperm from male CBAF1 mice (N=8) were incubated in either G-IVF+ or SpermFAST prior to IVF insemination. Fertilisation rates, embryo development, blastocyst cell numbers and DNA damage were assessed. Results: Incubation of human sperm in SpermFAST increased tyrosine phosphorylation (15.8% vs 9.5%, P[Formula: see text]0.05), hyperactive motility (38.3% vs 14.9%, P[Formula: see text]0.01), sperm binding (73.1% vs 47.7%, P[Formula: see text]0.01), while having no impact on sperm acrosomal status or oxidative DNA damage levels. Following IVF in the mouse, sperm incubated in SpermFAST increased fertilisation rates (94% vs 88%, P[Formula: see text]0.05), blastocyst total cell (92.2% vs 77.4%, P[Formula: see text]0.05), inner cell mass (14.9% vs 18.9%, P[Formula: see text]0.01) and epiblast cell numbers (3.7% vs 1.6%, P[Formula: see text]0.01), while the proportion of DNA damaged cells decreased in blastocysts (2.3% vs 4.8%, P[Formula: see text]0.001). Conclusion: Sperm function and fertilisation rates are improved when the sperm medium better mimics the environment of the female reproductive tract during natural conception. Improving IVF culture media to better meet the physiological needs of sperm could potentially improve outcomes following IVF.

Selection of a blastocyst for transfer is based on day5/6 morphology with day3 morphology only used if two blastocysts are equal in quality. Day3 assessments are based on cell number and fragmentation with embryos that are developmentally advanced ([Formula: see text]10 cells) being downgraded. After extended development, if [Formula: see text]10 cell embryos are the best quality they are selected for transfer. Thus, the aim of this study was to determine if cell number on day3 has an impact on embryo fate, euploidy rates and pregnancy rates. Data was retrospectively analysed from 269 women (average age 35.7) undergoing stimulated IVF treatment. Embryos were allocated to one of four categories based on day3 morphological score [Formula: see text]6 cells, 7-9 cells, 10-12 cells and [Formula: see text]13 cells. Results were correlated with embryo fate (vitrification/transfer) and pregnancy rates. After extended culture embryos with [Formula: see text]13 cells on day 3 were mostly likely to have undergone a fresh transfer (28.1%, p[Formula: see text]0.05) compared with embryos with 7-9 cells and 10-12 cells (13.7% and 14.0%, respectively). Embryos with [Formula: see text]6 cells were least likely to be transferred (5.2%) and vitrified (7.1%). Further, embryos with [Formula: see text]6 cells had the lowest pregnancy rate (12.5%) compared with all other groups (p[Formula: see text]0.05). No differences were observed for embryos with 7-9 cells, 10-12 cells and [Formula: see text]13 cells for vitrification rates (42.3%, 40.5% and 35.1%, respectively) or pregnancy rates (33.5%, 43.5% and 45.8%, respectively). Interestingly, the incidence of euploid embryos was increased in [Formula: see text]13 cell embryos (70.0%, p[Formula: see text]0.05) compared with [Formula: see text]6 cells, 7-9 cells and 10-12 cells (52.0%, 42.6% and 52.5%, respectively). Results from this study provide evidence that a review on cleavage stage embryo grading protocols which currently downgrades developmentally advanced embryos [Formula: see text]10 cells is warranted as these embryos do not appear to have a negative impact on pregnancy rates and can result in some of the best clinical outcomes.