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Background Early cancer detection could identify tumors at a time when outcomes are superior and treatment is less morbid. This prospective case-control sub-study (from NCT02889978 and NCT03085888) assessed the performance of targeted methylation analysis of circulating cell-free DNA (cfDNA) to detect and localize multiple cancer types across all stages at high specificity. Participants and methods The 6689 participants [2482 cancer (>50 cancer types), 4207 non-cancer] were divided into training and validation sets. Plasma cfDNA underwent bisulfite sequencing targeting a panel of >100 000 informative methylation regions. A classifier was developed and validated for cancer detection and tissue of origin (TOO) localization. Results Performance was consistent in training and validation sets. In validation, specificity was 99.3% [95% confidence interval (CI): 98.3% to 99.8%; 0.7% false-positive rate (FPR)]. Stage I–III sensitivity was 67.3% (CI: 60.7% to 73.3%) in a pre-specified set of 12 cancer types (anus, bladder, colon/rectum, esophagus, head and neck, liver/bile-duct, lung, lymphoma, ovary, pancreas, plasma cell neoplasm, stomach), which account for ∼63% of US cancer deaths annually, and was 43.9% (CI: 39.4% to 48.5%) in all cancer types. Detection increased with increasing stage: in the pre-specified cancer types sensitivity was 39% (CI: 27% to 52%) in stage I, 69% (CI: 56% to 80%) in stage II, 83% (CI: 75% to 90%) in stage III, and 92% (CI: 86% to 96%) in stage IV. In all cancer types sensitivity was 18% (CI: 13% to 25%) in stage I, 43% (CI: 35% to 51%) in stage II, 81% (CI: 73% to 87%) in stage III, and 93% (CI: 87% to 96%) in stage IV. TOO was predicted in 96% of samples with cancer-like signal; of those, the TOO localization was accurate in 93%. Conclusions cfDNA sequencing leveraging informative methylation patterns detected more than 50 cancer types across stages. Considering the potential value of early detection in deadly malignancies, further evaluation of this test is justified in prospective population-level studies.

In the Circulating Cell-free Genome Atlas (NCT02889978) substudy 1, we evaluate several approaches for a circulating cell-free DNA (cfDNA)-based multi-cancer early detection (MCED) test by defining clinical limit of detection (LOD) based on circulating tumor allele fraction (cTAF), enabling performance comparisons. Among 10 machine-learning classifiers trained on the same samples and independently validated, when evaluated at 98% specificity, those using whole-genome (WG) methylation, single nucleotide variants with paired white blood cell background removal, and combined scores from classifiers evaluated in this study show the highest cancer signal detection sensitivities. Compared with clinical stage and tumor type, cTAF is a more significant predictor of classifier performance and may more closely reflect tumor biology. Clinical LODs mirror relative sensitivities for all approaches. The WG methylation feature best predicts cancer signal origin. WG methylation is the most promising technology for MCED and informs development of a targeted methylation MCED test.

A middle to late Pleistocene sedimentary sequence in the upper Las Vegas Wash, north of Las Vegas, Nevada, has yielded the largest open-site Rancholabrean vertebrate fossil assemblage in the southern Great Basin and Mojave Deserts. Recent paleontologic field studies have led to the discovery of hundreds of fossil localities and specimens, greatly extending the geographic and temporal footprint of original investigations in the early 1960s. The significance of the deposits and their entombed fossils led to the preservation of 22,650 acres of the upper Las Vegas Wash as Tule Springs Fossil Beds National Monument. These discoveries also warrant designation of the assemblage as a local fauna, named for the site of the original paleontologic studies at Tule Springs. The large mammal component of the Tule Springs local fauna is dominated by remains of Mammuthus columbi as well as Camelops hesternus , along with less common remains of Equus (including E. scotti ) and Bison . Large carnivorans including Canis dirus , Smilodon fatalis , and Panthera atrox are also recorded. Micromammals, amphibians, lizards, snakes, birds, invertebrates, plant macrofossils, and pollen also occur in the deposits and provide important and complementary paleoenvironmental information. The fauna occurs within the Las Vegas Formation, an extensive and stratigraphically complex sequence of groundwater discharge deposits that represent a mosaic of desert wetland environments. Radiometric and luminescence dating indicates the sequence spans the last ∼570 ka, and records hydrologic changes in a dynamic and temporally congruent response to northern hemispheric abrupt climatic oscillations. The vertebrate fauna occurs in multiple stratigraphic horizons in this sequence, with ages of the fossils spanning from ∼100 to ∼12.5 ka.

Room temperature optical illumination of NV- imbibed single crystal diamonds with a 532 nm laser produces (13)C polarization enhancements up to 200 times greater than that of the thermal equilibrium value at 7.05 T. We report high field NV- mediated (13)C polarization as a function of the number and type (NV- and P1) of defects in commercially available diamonds. Surprisingly, both positive and negative (13)C polarizations are observed depending on the orientation of the crystal with respect to the external magnetic field and the electric field vector of the optical illumination. The data reported herein cannot be explained by a previously proposed mechanism.

Background Spartalizumab (PDR001) is an anti-PD-1 monoclonal antibody that is been explored in different tumor types. Preliminary safety and efficacy results from Groups B (B) and C (C) of a phase Ib study investigating spartalizumab with platinum-based chemotherapy in PD-L1 unselected advanced NSCLC (NCT03064854) are reported here. Methods Treatment-naive, stage IIIB/IV NSCLC, EGFR/ALK/ROS-1(-) pts, PS 0-1, were assigned to B (non-squamous NSCLC) or C (squamous/non-squamous NSCLC) for 4 cycles (21 day/cycle) of spartalizumab (initial dose – 300 mg Q3W) + pemetrexed 500 mg/m2 + cisplatin 75 mg/m2 followed by maintenance spartalizumab + pemetrexed (in B), or 4 cycles of spartalizumab (initial dose – 300 mg Q3W) + paclitaxel 200 mg/m2 + carboplatin AUC 6 followed by maintenance with spartalizumab (in C). Primary endpoints are recommended dose for expansion (RDE) and ORR by investigator assessment. Results At data cutoff of 15 May 2019, 38 pts in B and 33 pts in C were treated; 14 pts (36.8%) in B and 4 pts (12.1%) in C are still receiving treatment. Primary reason for discontinuation was progressive disease: 13 of 38 pts (34.2%) in B and 25 of 33 pts (75.8%) in C. DLTs were grade 4 hyponatremia and grade 2 posterior reversible encephalopathy syndrome in B (2 pts) and grade 4 neutropenic colitis in C (1 pt). RDE of spartalizumab in combination with platinum-based chemotherapy for both groups was 300 mg Q3W. Most common AEs (≥50%, any grade) were nausea (73.7%) and neutropenia (50%) in B; neutropenia (57.6%) and anemia (54.5%) in C. ORR and complete response rate were 50% and 5.3% in B, and 51.5% and 3% in C. Median DOR (months; 95% CI) by investigator assessment was 13.8 (4.9, NE) in B and 8.2 (5.1, 15.4) in C. Median PFS (months; 95% CI) by investigator assessment was 10.4 (5.4, 15.0) in B and 6.3 (4.1, 10.1) in C. Responses were observed at different PD-L1 expression levels with higher rate in PD-L1 tumor proportion score (TPS) ≥ 50%. Assessment of additional immune biomarkers is ongoing. Conclusions RDE of spartalizumab in combination with platinum-based chemotherapy is 300 mg Q3W. Safety profile and preliminary clinical activity are consistent with prior experience with spartalizumab, other immune checkpoint inhibitors and chemotherapy treatments administered. Clinical trial identification NCT03064854. Editorial acknowledgement Shiva Krishna Rachamadugu, Novartis Healthcare Pvt Ltd, Hyderabad, India. Legal entity responsible for the study Novartis Pharmaceuticals Corporation. Funding Novartis Pharmaceuticals Corporation. Disclosure A. Santoro: Advisory / Consultancy: BMS, Servier, Gilead, Pfizer, Eisai, Bayer, Msd, Arqule; Speaker Bureau / Expert Testimony: Takeda, BMS, Roche, AbbVie, Amgen, Celgene, Servier, Gilead, Astrazeneca, Pfizer, Arqule, Lilly, Sandoz, Eisai, Novartis, Bayer, MSD. P. Garrido Lopez: Advisory / Consultancy: Roche, MSD, BMS, Boerhinger Ingelheim, Pfizer, AbbVie, Guardant Health, Novartis, Lilly, AstraZeneca, Janssen, Sysmex, Blueprint Medicines, Takeda; Speaker Bureau / Expert testimony: Roche, MSD, BMS, Pfizer, Novartis, Boerhinger Ingelheim, Rovi; Research grant / Funding (institution), For Clinical Trails: Roche, MSD, BMS, Takeda, Lilly, Pfizer, Novartis, PharmaMar, Celgene, Sanofi, GSK, Theradex Oncology, BluePrint Medicines; Research grant / Funding (institution), For Contracted Research: Guardant Health, Sysmex. D.S.W. Tan: Honoraria (self): Merck, Pfizer, Novartis, Boehringer Ingelheim, Roche, Takeda ; Advisory / Consultancy: Novartis, Bayer, Boehringer Ingelheim, Celgene, AstraZeneca, Eli-lily, Loxo; Research grant / Funding (self): Novartis, AstraZeneca, GlaxoSmithKline, Bayer, Pfizer ; Travel / Accommodation / Expenses: Merck, Pfizer, Novartis, Boehringer Ingelheim, Roche,Takeda . L. Paz-Ares: Honoraria (self): Roche, Novartis, Pfizer, Lilly, BMS, MSD, Merck, Boehringer Ing., AstraZeneca, Amgen, Sanofi, PharmaMar, Takeda; Leadership role: Genomica. F. Shepherd: Honoraria (self): Novartis, AstraZeneca, Roche, BMS, Merck, EMD Serono; Advisory / Consultancy: Novartis, AstraZeneca, Roche, BMS, Merck, EMD Serono; Shareholder / Stockholder / Stock options: AstraZeneca. A. Bearz: Advisory / Consultancy: Takeda; Boehringer Ingelheim, Roche , MSD, Novartis; Speaker Bureau / Expert testimony: Takeda; Boehringer Ingelheim, Roche , MSD; Pfizer; Research grant / Funding (institution): AstraZeneca. F. Barlesi: Honoraria (self): AstraZeneca, Bayer, Bristol-Myers Squibb, Boehringer–Ingelheim, Eli Lilly Oncology, F. Hoffmann–La Roche Ltd, Novartis, Merck, MSD, Pierre Fabre, Pfizer and Takeda; Research grant / Funding (institution): AbbVie, ACEA, Amgen, AstraZeneca, Bayer, Bristol-Myers Squibb, Boehringer–Ingelheim, Eisai, Eli Lilly Oncology, F. Hoffmann–La Roche Ltd, Genentech, Ipsen, Ignyta, Innate Pharma, Loxo, Novartis, MedImmune, Merck, MSD, Pierre Fabre, Pfizer, Sanofi-Aventis; Non-remunerated activity/ies: Principal Investigator for AstraZeneca, BMS, Merck, Pierre Fabre and F. Hoffmann-La Roche, Ltd, sponsored trials (or ISR). J.F. Vansteenkiste: Honoraria (self), Advisory / Consultancy: AstraZeneca, Boehringer Ingelheim, Eli Lilly, MSD, Roche, Apotex; Research grant / Funding (institution): MSD. T.M. Kim: Research grant / Funding (institution): Alpha Biopharma, AstraZeneca/MedImmune, Hanmi, Janssen, Merus, Mirati Therapeutics, MSD, Novartis, ONO Pharmaceutical, Pfizer, Roche/Genentech, Takeda, TP Therapeutics, Xcovery, and Yuhan ; Research grant / Funding (self): AstraZeneca ; Non-remunerated activity/ies, Advisory / Consultancy: AstraZeneca, Novartis, Sanofi, and Bayer . T.R. Overbeck: Advisory / Consultancy, Consulting fees: AstraZeneca, BMS, Boehringer Ingelheim, Eli Lilly, Medac, MSD, Novartis, Roche/Genentech; Research grant / Funding (institution), sponsored research agreements: AstraZeneca, Eli Lilly, Roche/Genentech, Sanofi-Aventis; Travel / Accommodation / Expenses, Travel Support: AstraZeneca, Boehringer Ingelheim, Eli Lilly and Roche/Genentech. I.I. Rybkin: Research grant / Funding (institution): Merck, AbbVie, ARMO Biosciences, Beyond Spring Pharmaceutical, Bristol-Myers Squibb, Novartis, Mirati Therapeutical Inc., Nilogen, Inc., Pfizer, Polaris Group, Syndax Inc., Xcovery Inc., BerGenBio AS, Incyte Corporation, AstraZeneca, Amgen; Advisory / Consultancy: AstraZeneca. E. Felip: Advisory / Consultancy: AbbVie, AstraZeneca, Blue Print Medicines, Boehringer Ingelheim, Bristol-Myers Squibb, Celgene, Eli Lilly, Guardant Helath, Janssen, Medscape, Merck Kgaa, Merck Sharp & Dohme, Novartis, Pfizer, Roche, Takeda, Touchtime; Speaker Bureau / Expert Testimony: AbbVie, AstraZeneca, Boehringer Ingelheim, Bristol-Myers Squibb, Eli Lilly, Merck Kgaa, Merck Sharp & Dohme, Novartis, Pfizer, Roche, Takeda; Research Grant / Funding (Self): Fundacion Merck Salud Grant For Oncology Innovation Emd Serono. W. Zhou: Full / Part-time employment: Novartis. L. Santarpia: Full / Part-time employment: Novartis. S. Eddy: Shareholder / Stockholder / Stock options, Full / Part-time employment: Novartis. All other authors have declared no conflicts of interest.

In this study, placing parallel-connected baffles in the vicinity of the inlet was proposed to improve hydraulic conditions for enhancing TSS (total suspended solids) removal. The purpose of the retrofit baffle design is to divide the large and fast inflow into smaller and slower flows to increase flow uniformity. This avoids short-circuiting and increases residence time in the sedimentation basin. The newly proposed parallel-connected baffle configuration was assessed in the laboratory by comparing its TSS removal performance and the optimal flow residence time with those from the widely used series-connected baffles. The experimental results showed that the parallel-connected baffles outperformed the series-connected baffles because it could disperse flow faster and in less space by splitting the large inflow into many small branches instead of solely depending on flow internal friction over a longer flow path, as was the case under the series-connected baffles. Being able to dampen faster flow before entering the sedimentation basin is critical to reducing the possibility of disturbing any settled particles, especially under high inflow conditions. Also, for a large sedimentation basin, it may be more economically feasible to deploy the proposed parallel retrofit baffle in the vicinity of the inlet than series-connected baffles throughout the entire settling basin.

Dendritic spines represent the major site of neuronal activity in the brain; they serve as the receiving point for neurotransmitters and undergo rapid activity-dependent morphological changes that correlate with learning and memory. Using a combination of homozygosity mapping and next-generation sequencing in two consanguineous families affected by nonsyndromic autosomal-recessive intellectual disability, we identified truncating mutations in formin 2 (FMN2), encoding a protein that belongs to the formin family of actin cytoskeleton nucleation factors and is highly expressed in the maturing brain. We found that FMN2 localizes to punctae along dendrites and that germline inactivation of mouse Fmn2 resulted in animals with decreased spine density; such mice were previously demonstrated to have a conditioned fear-learning defect. Furthermore, patient neural cells derived from induced pluripotent stem cells showed correlated decreased synaptic density. Thus, FMN2 mutations link intellectual disability either directly or indirectly to the regulation of actin-mediated synaptic spine density.

The corpus callosum is the principal cerebral commissure connecting the right and left hemispheres. The development of the corpus callosum is under tight genetic control, as demonstrated by abnormalities in its development in more than 1,000 genetic syndromes. We recruited more than 25 families in which members affected with corpus callosum hypoplasia (CCH) lacked syndromic features and had consanguineous parents, suggesting recessive causes. Exome sequence analysis identified C12orf57 mutations at the initiator methionine codon in four different families. C12orf57 is ubiquitously expressed and encodes a poorly annotated 126 amino acid protein of unknown function. This protein is without significant paralogs but has been tightly conserved across evolution. Our data suggest that this conserved gene is required for development of the human corpus callosum.

The tRNA splicing endonuclease is a highly evolutionarily conserved protein complex, involved in the cleavage of intron-containing tRNAs. In human it consists of the catalytic subunits TSEN2 and TSEN34, as well as the non-catalytic TSEN54 and TSEN15. Recessive mutations in the corresponding genes of the first three are known to cause pontocerebellar hypoplasia (PCH) types 2A-C, 4, and 5. Here, we report three homozygous TSEN15 variants that cause a milder version of PCH2. The affected individuals showed progressive microcephaly, delayed developmental milestones, intellectual disability, and, in two out of four cases, epilepsy. None, however, displayed the central visual failure seen in PCH case subjects where other subunits of the TSEN are mutated, and only one was affected by the extensive motor defects that are typical in other forms of PCH2. The three amino acid substitutions impacted the protein level of TSEN15 and the stoichiometry of the interacting subunits in different ways, but all resulted in an almost complete loss of in vitro tRNA cleavage activity. Taken together, our results demonstrate that mutations in any known subunit of the TSEN complex can cause PCH and progressive microcephaly, emphasizing the importance of its function during brain development.

Nested R-groups are fundamental to patent Markush structure’s complexity, yet there have not been many recent publications in this field. We have developed a web application, called iMarVis, which provides chemists with the ability to navigate nested R-groups and study selected subsets of chemical space. In order to accurately capture R-group hierarchy, a new underlying numbering system was created. This numbering system also facilitates additional Markush structure analysis, including chemical space size calculation and enumeration. This article describes the new numbering system in detail and highlights the importance of understanding and depicting the R-group hierarchy.

Proposed explanations for the terminal Pleistocene large mammal extinction event in North America include climate warming and/or cooling, overhunting by early humans, disease, and bolide detonation or impact. A key assumption common to all these scenarios is that large mammals present in North America near the end of the Pleistocene were also present in similar abundance, with similar geographic distributions, during earlier, equally severe periods of climate change (e.g., ∼130 ka BP). This assumption is challenged here. An important difference in the latest Pleistocene was the profusion and geographic extent of the genus Bison , particularly in the American West. During the late Pleistocene, south of the glacial ice, the species Bison antiquus was more widely distributed and present in greater profusion than earlier species such as the larger B. latifrons . The increased abundance of these large, aggressive, herd-dwelling ruminants in the late Pleistocene constitutes a critical difference between this time period and earlier, similarly intense interglacials. Extinction scenarios for Pleistocene North America should avoid assuming a relatively static long-term faunal component, and account for the impacts of non-human immigrant species on natives, particularly when immigration events are close in time and space with climate changes.

The Diamond Valley Lake local fauna from southwestern Riverside County, California is characterized by a classic suite of well-preserved late Pleistocene (Rancholabrean NALMA) vertebrates, including over 100,000 identifiable fossils representing more than 105 vertebrate, invertebrate and plant taxa from 2646 localities. The fauna is the largest open-environment, non-asphaltic late Pleistocene assemblage known from the American southwest. Located within the northern Peninsular Range physiographic province of southern California, the Diamond and Domenigoni Valleys contain bedded silts and clays intercalated with coarse-grained channel fill representing a braided stream environment. These fluvial sediments, yielding AMS dates from ∼19 ka to ∼13 ka, unconformably truncate older silts, clays and an organic black clay at depth. The clay is lacustrine in origin, with AMS dates from ∼46 ka to ∼41 ka. Numerous diagnostic vertebrate remains occur in both of these sediment packages. The Diamond Valley Lake local fauna constitutes a valuable source of new data on the relative density and diversity of late Pleistocene species from a geographic area where such data are largely absent. The assemblage differs dramatically in preservation and composition from other late Pleistocene coastal and desert localities in southern California.

A simple synthetic procedure for preparing biocompatible superparamagnetic Prussian blue (PB) nanoparticles is reported. The stability, cytotoxicity and ability for PB nanoparticles to penetrate cells are investigated. The potential of using PB nanoparticles as the T1-weighted MRI contrast agent is demonstrated.

The concept of ovarian reserve describes the natural oocyte endowment and is closely associated with female age, which is the single most important factor influencing reproductive outcome. Fertility potential first declines after the age of 30 and moves downward rapidly thereafter, essentially reaching zero by the mid-40s. Conceptions beyond this age are exceedingly rare, unless oocytes obtained from a younger donor are utilised. How best to estimate ovarian reserve clinically remains controversial. Passive assessments of ovarian reserve include measurement of serum follicle stimulating hormone (FSH), oestradiol (E(2)), anti-Müllerian hormone (AMH), and inhibin-B. Ultrasound determination of antral follicle count (AFC), ovarian vascularity and ovarian volume also can have a role. The clomiphene citrate challenge test (CCCT), exogenous FSH ovarian reserve test (EFORT), and GnRH-agonist stimulation test (GAST) are provocative methods that have been used to assess ovarian reserve. Importantly, a patient's prior response to gonadotropins also provides highly valuable information about ovarian function. Regarding prediction of reproductive outcome, in vitro fertilisation (IVF) experience at our centres and elsewhere has shown that some assessments of ovarian reserve perform better than others. In this report, these tests are discussed and compared; we also present practical strategies to organise screening as presently used at our institutions. Experimental challenges to the long-held tenet of irreversible ovarian ageing are also introduced and explored. While pregnancy rates after IVF are influenced by multiple (non-ovarian) factors including in vitro laboratory conditions, semen parameters, psychological stress and technique of embryo transfer, predicting response to gonadotropin treatment nevertheless remains an important aim in the evaluation of the couple struggling with infertility.

In this brief communication we show several examples demonstrating the usefulness of non-resonant transmission line probes. Although rarely seen in the literature and typically limited to zero external field experiments on ferromagnetic materials we have found that this type of probe is perfectly capable of a variety of other nuclear magnetic resonance experiments including cross-polarization.

The sea urchin embryo follows a relatively simple cell behavioral sequence in its gastrulation movements. The embryo reaches the gastrula stage as a spherical monolayer of cells. To form the mesoderm, primary mesenchyme cells ingress by delaminating from the vegetal plate, crossing the basal lamina and moving into the central blastocoelar cavity. These cells then migrate along the basal lamina lining the blastocoel and eventually manufacture the skeleton. The presumptive secondary mesenchyme and endoderm invaginate as a tubular sheet of cells from the vegetal pole of the embryo. The archenteron extends across the blastocoel until its tip touches and attaches to the opposite side of the blastocoel. Secondary mesenchyme cells, originally at the tip of the archenteron, differentiate to form a variety of structures including coelomic pouches, esophageal muscles, pigment cells, and other cell types. The endoderm fuses with an invagination of the ventral ectoderm (the stomodaem), to form the mouth and complete the process of gastrulation. A number of experiments have established that these simple morphogenetic movements are accompanied by a number of cell adhesion changes plus a series of cell-cell interactions that provide spatial, temporal, and scalar information to cells of the mesoderm and endoderm. The requirement for cell signaling has been demonstrated by manipulative experiments where it has been shown that axial, temporal, spatial, and scalar information is obtained by mesoderm and endoderm from other embryonic cells. That information governs pattern formation and subsequent adhesive changes. This review describes the adhesion changes and the signaling that characterizes this early morphogenesis.

In this paper, we argue that ambiguity, that is people’s inability to judge likelihood of risk, is a significant contributor to aversion to spyware risk. Since no objective or authoritative estimates of risk likelihood are available, but users still make decision, it must be the case that user’s judgments of risk likelihood are reflected in their decision weights. Using the theory and methods developed in the field of decision theory, we plan to conduct an experiment to a) assess the separate contributions of standard risk aversion and aversion to ambiguity to overall risk and b) examine whether peoples traits (optimism/pessimism, tolerance for ambiguity) and perception of information explain the patterns in the parameters corresponding to risk and ambiguity functions.

In this manuscript, the effects of fetal bovine serum (FBS) on activation of mouse macrophages through the p73 lipopolysaccharide (LPS) receptor have been evaluated. In confirmation of earlier published studies, FBS will significantly potentiate the ability of LPS to activate macrophages to produce nitric oxide (NO). Evidence that this potentiating effect of FBS is mediated primarily by an interaction with LPS is provided by data showing that the stimulating effects of a hamster IgM monoclonal antibody to the p73 LPS receptor are significantly suppressed under identical conditions of FBS addition. The presence of FBS enhances both the kinetics of LPS-induced NO production and delays the induction of MAb5D3-induced NO production. The data establish that the ability of FBS to reduce MAb5D3-initiated NO production can only be manifest during the first 4-6 h following activation with antibody. Similarly, the ability of polymyxin B (which does not affect MAb5D3 activity) to inhibit LPS-dependent macrophage activation is also only effective during the first 4-6 h of stimulation, suggesting a parallel kinetic profile in the activation of the inducible NO synthase by these related activators. These studies provide data which suggest a dual role for serum factors in LPS-dependent macrophage activation, a direct effect of FBS on LPS which potentiates its immunostimulatory activity, and a secondary down-regulation effect which is manifest at the level of the macrophage.

OBJECTIVE: Typically, blastocysts are biopsied (bx) and vitrified while awaiting preimplantation genetic screening (PGS) results. Sometimes blastocysts receive ‘‘no read’’ post PGS, leaving vitrified blastocysts with no genetic information. Furthermore, patients can desire PGS on previously frozen blastocysts which PGS was not performed. These scenarios require blastocysts to be warmed, bx’d, vitrified, and warmed again. The effect of double freezing and double warming on outcomes is unknown. DESIGN: Retrospective. MATERIALS ANDMETHODS: Patients undergoing blastocysts bx with PGS were included. Patients were divided into 2 groups, group 1 patients transferred (ETd) blastocysts that were vitrified and warmed once. Group 2 patients ETd blastocysts that were frozen, warmed, bx’d, vitrified, and rewarmed. RESULTS: A total of 75 patients, 36.7 3.9 years, and 17 patients, 35.3 4.9 years, were included in group 1 and 2, respectively (NS). Blastocyst warm survival rates between group 1 (101 of 103, 98.1%) and 2 (20 of 23, 87.0%) was significant (0.0420). All 75 patients received an ET from group 1, while 16 patients received an ET in group 2. The average number ET’d was 1.3 0.5 and 1.3 0.6 for group 1 and 2, respectively (NS). Chemical pregnancies (54 of 75, 72.0%) and (10 of 16, 62.5%), clinical gestations (48 of 75, 64.0%) and (9 of 16, 56.3%), clinical pregnancies (45 of 75, 60.0%) and (8 of 16, 50.0%), and implantation rates (65 of 100, 65.0%) and (11 of 20, 55.0%), were not significant between groups 1 and 2, respectively. Of the 61 clinical heartbeats from group 1, 31 were live births, 1 still birth, 25 ongoing, 3 therapeutic abortions, and 1 miscarriage. Of the 10 clinical heartbeats from group 2, 9 are live births and 1 ongoing. CONCLUSION: Although it is unconventional to thaw, bx, revitrify, and rewarm a blastocysts for ET, our results indicate that as long as the blastocyst survives the warming, outcomes are unaffected. Supported by: REACh, Charlotte, NC.

Summary Ossification of the lateral cartilage of the distal hind phalanx of an individual of Equus “occidentalis” from the Rancho La Brea asphalt deposits is described. This is the first known report of “sidebone” among ancient horses.