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Mercury (Hg) export from glacierized watersheds is poorly understood, with very few studies worldwide on Hg concentration and speciation in glacier snow, ice, and meltwater, and on Hg fluxes to downstream freshwater and coastal ecosystems. In addition to bedrock-derived geogenic Hg, glaciers may be releasing legacy accumulations of natural and anthropogenically-sourced Hg trapped in glacier ice melt each summer season. Our prior work showed that a glacierized stream in southeast Alaska had the highest reported flux of inorganic mercury of all known non-mining impacted streams, highlighting the strong ability of glaciers to mobilize the trace metal. Here we present data on Hg concentrations, speciation, partitioning, and fluxes from two more glacierized watersheds (Herbert and Mendenhall Rivers), and we compare them to an adjacent non-glacierized, forested-wetland stream (Peterson Creek). Results show that the glacierized streams carried Hg largely in the particulate form, whereas the forested-wetland stream carried it largely in the filtered fraction, at 20 fold higher concentration than in the glacierized streams, and with a higher percent of Hg in its methylated form. Yet, considering the higher water and sediment yields (as mass per watershed area per year) of the glacierized streams during the summer melt season, the yield of total Hg (unfiltered) from the Mendenhall glacier was approximately 80 times higher than from the Herbert glacier and 50 times higher than in Peterson Creek and presents the highest watershed yields of total Hg and methyl-Hg reported in the literature to date. Incongruous yields out of the two glacierized streams can likely be explained by differences in underlying bedrock geology. Based on the sediments entrained in Mendenhall meltwater, the late Paleozoic to Paleocene metasedimentary and volcanic rocks being eroded in the terminal 3 km of the Mendenhall are elevated above mean crustal concentrations by at least 4–17 fold. Differences in speciation between the glacierized and non-glacierized streams are likely accounted for by glacial and watershed geochemical conditions that variably promote Hg methylation.

Sexually transmitted infections are of major concern to reproductive specialists. Heading the list are human immunodeficiency virus types 1 and 2 and hepatitis B and C viruses. These pathogens, which may cause incurable chronic infections, can be transmitted through assisted reproductive technologies and from infected mothers to the fetus or newborn. This document replaces the document of the same name last published in 2013 (Fertil Steril 2013;99:340-6).

SummaryAt the moment of union in fertilization, sperm and oocyte are transcriptionally silent. The ensuing onset of embryonic transcription (embryonic genome activation [EGA]) is critical for development, yet its timing and profile remain elusive in any vertebrate species. We here dissect transcription during EGA by high-resolution single-cell RNA sequencing of precisely synchronized mouse one-cell embryos. This reveals a program of embryonic gene expression (immediate EGA [iEGA]) initiating within 4 h of fertilization. Expression during iEGA produces canonically spliced transcripts, occurs substantially from the maternal genome, and is mostly downregulated at the two-cell stage. Transcribed genes predict regulation by transcription factors (TFs) associated with cancer, including c-Myc. Blocking c-Myc or other predicted regulatory TF activities disrupts iEGA and induces acute developmental arrest. These findings illuminate intracellular mechanisms that regulate the onset of mammalian development and hold promise for the study of cancer.

In human embryos, the initiation of transcription (embryonic genome activation [EGA]) occurs by the eight-cell stage, but its exact timing and profile are unclear. To address this, we profiled gene expression at depth in human metaphase II oocytes and bipronuclear (2PN) one-cell embryos. High-resolution single-cell RNA sequencing revealed previously inaccessible oocyte-to-embryo gene expression changes. This confirmed transcript depletion following fertilization (maternal RNA degradation) but also uncovered low-magnitude upregulation of hundreds of spliced transcripts. Gene expression analysis predicted embryonic processes including cell-cycle progression and chromosome maintenance as well as transcriptional activators that included cancer-associated gene regulators. Transcription was disrupted in abnormal monopronuclear (1PN) and tripronuclear (3PN) one-cell embryos. These findings indicate that human embryonic transcription initiates at the one-cell stage, sooner than previously thought. The pattern of gene upregulation promises to illuminate processes involved at the onset of human development, with implications for epigenetic inheritance, stem-cell-derived embryos, and cancer.

In this study, a stream from a glacially dominated watershed and one from a wetland, temperate forest dominated watershed in southeast Alaska were continuously monitored for turbidity and fluorescence from dissolved organic matter (FDOM) while grab samples for unfiltered (UTHg), particulate (PTHg), and filtered mercury (FTHg) where taken over three 4-day periods (May snowmelt, July glacial melt, and September rainy season) during 2010. Strong correlations were found between FDOM and UTHg concentrations in the wetland, temperate forest watershed (r2=0.81), while turbidity and UTHg were highly correlated in the glacially dominated watershed (r2=0.82). Both of these parameters (FDOM and turbidity) showed stronger correlations than concentration-discharge relationships for UTHg (r2=0.55 for glacial stream, r2=0.38 for wetland/forest stream), thus allowing for a more precise determination of temporal variability in UTHg concentrations and fluxes. The association of mercury with particles and dissolved organic matter (DOM) appears to depend on the watershed characteristics, such as physical weathering and biogeochemical processes regulating mercury transport. Thus employing watershed-specific proxies for UTHg (such as FDOM and turbidity) can be effective for quantifying mercury export from watersheds with variable landcover. The UTHg concentration in the forest/wetland stream was consistently higher than in the glacial stream, in which most of the mercury was associated with particles; however, due to the high specific discharge from the glacial stream during the melt season, the watershed area normalized flux of mercury from the glacial stream was 3-6 times greater than the wetland/forest stream for the three sampling campaigns. The annual specific flux for the glacial watershed was 19.9gUTHgkm-2y-1, which is higher than any non-mining impacted stream measured to date. This finding indicates that glacial watersheds of southeast Alaska may be important conduits of total mercury to the Gulf of Alaska.

Due to anatomical and physiological similarities to humans, the common marmoset (Callithrix jacchus) is an ideal organism for the study human diseases. Researchers are currently leveraging genome-editing technologies such as CRISPR/Cas9 to genetically engineer marmosets for the in vivo biomedical modeling of human neuropsychiatric and neurodegenerative diseases. The genome characterization of these cell lines greatly reinforces these transgenic efforts. It also provides the genomic contexts required for the accurate interpretation of functional genomics data. We performed haplotype-resolved whole-genome characterization for marmoset ESC line cj367 from the Wisconsin National Primate Research Center. This is the first haplotype-resolved analysis of a marmoset genome and the first whole-genome characterization of any marmoset ESC line. We identified and phased single-nucleotide variants (SNVs) and Indels across the genome. By leveraging this haplotype information, we then compiled a list of cj367 ESC allele-specific CRISPR targeting sites. Furthermore, we demonstrated successful Cas9 Endonuclease Dead (dCas9) expression and targeted localization in cj367 as well as sustained pluripotency after dCas9 transfection by teratoma assay. Lastly, we show that these ESCs can be directly induced into functional neurons in a rapid, single-step process. Our study provides a valuable set of genomic resources for primate transgenics in this post-genome era.

Computer-automated time-lapse analysis has been shown to improve embryo selection by providing quantitative and objective information to supplement traditional morphology. In this multi-centre study, the relationship between such computer-derived outputs (High, Medium, Low scores), embryo implantation and clinical pregnancy were examined. Data were collected from six clinics, including 205 patients whose embryos were imaged by the Eeva TM System. The Eeva scores were blinded and not considered during embryo selection. Embryos with High and Medium scores had significantly higher implantation rates than those with Low scores (37% and 35% versus 15%; P P = 0.0004). Similar trends in implantation rates were observed in different IVF centres each using their own protocols. Further analysis revealed that patients with at least one High embryo transferred had significantly higher clinical pregnancy rates than those with only Low embryos transferred (51% versus 34%; P = 0.02), although patients' clinical characteristics across groups were comparable. These data, together with previous research and clinical studies, confirm that computer-automated Eeva scores provide valuable information, which may improve the clinical outcome of IVF procedures and ultimately facilitate the trend of single embryo selection.

Objective We examined embryo development potential and pregnancy outcomes from blastocysts that developed from 0PNpn, 1PB zygotes. Design A retrospective study in private IVF laboratory. Materials and Methods Over the course of twenty-four months, 68 blastocysts developed from mature oocytes that did not show visual signs of 2 pronuclei at approximently18hrs post-insemination by ICSI. The 0PN1PB embryos were cultured in Single Continuous Culture-NX (Irvine Scientific®) separately from the normally fertilized (2PN) zygote cohort and monitored accordingly. Any blastocysts obtained from 0PN1PB zygotes were either biopsied and/or frozen on day 5, 6 or 7 of embryo development. Results January 2017-December 2018, 68 blastocysts obtained from 0PN1PB zygotes represented approximately 1% of the total number of embryos cultured in our laboratory. Of the 68 blastocysts, 51were biopsied and subsequently tested by NexGen sequencing resulting in 23 (45%) of which were deemed euploid. 12 embryo transfers with euploid embryos were performed (11 frozen, 1 fresh) resulting in 5 (42%) with positive clinical pregnancies and 3 live births. Positive clinical pregnancy was defined cardiac activity visualized via ultrasound at 7 weeks gestation. The frequency of chromosomes that were chromosomally abnormal as indicated by PGT-A was also retrospectively evaluated which resulted in no correlation of a particular set of chromosomes Conclusion The result of 0PN1PB after ICSI is an artifact of every IVF program. The inefficiency of these embryos to develop into a blastocyst is well known. While the population of embryos produced from 0PN1PBs is considerably low, the minimal extra work and monitoring required does not outweigh the potential benefits of culturing these embryos to the blastocyst stage. Although our study numbers are small, this retrospective observation indicates that these embryos do have the potential to develop into a chromosomally normal blastocyst which can result in a live birth. Embryo transfer data is further limited by preference in selecting embryos which have resulted from 2PN zygotes versus those from 0PN1PB zygotes. However, by utilizing PGT-A technology, our study further validates that these embryos have similar euploid outcomes compared to those blastocysts which have developed from 2PN zygotes. With proper embryo culture organization, these embryos can be monitored effortlessly and become part of any IVF routine. Funding None Disclosures None