Search

Design of engineered combustion equipment normally involves laborious “build and try” designs to identify the best possible configuration. The number of design iterations can be reduced with engineering experience of what might work. The expensive cut-and-try approach can be improved using computational aided engineering tools coupled with optimization techniques to find the optimal design. For example, the “best” air duct configuration with the lowest pressure loss and smallest fan size for an air-fed biomass gasifier may take several weeks using the standard computational fluid dynamics (CFD) “cut and try” approach. Alternatively, coupling an efficient design optimization algorithm with an existing CFD model can reduce the time to find the best design by more than 50% and can allow the engineer to examine more design options than possible using the “cut-and-try” approach. Combining an efficient optimization algorithm with an existing CFD model of a biomass gasifier to find the “optimal” design is the focus of this work. Shape optimization has been performed by combining the optimization tool Sculptor® with the commercial CFD code STARCCM+. This work illustrates how the “linked” approach is used to examine design factors to optimize an entrained flow biomass gasifier to improve overall system performance in a methodical comprehensive fashion.

Summary Hematopoiesis is arguably one of the best understood stem cell systems; however, significant challenges remain to reach a consensus understanding of the lineage potential, heterogeneity, and relationships of hematopoietic stem and progenitor cell populations. To gain new insights, we performed quantitative analyses of mature cell production from hematopoietic stem cells (HSCs) and multiple hematopoietic progenitor populations. Assessment of the absolute numbers of mature cell types produced by each progenitor cell revealed a striking erythroid dominance of all myeloid-competent progenitors assessed, accompanied by strong platelet reconstitution. All populations with myeloid potential also produced robust numbers of red blood cells and platelets in vivo. Clonal analysis by single-cell transplantation and by spleen colony assays revealed that a significant fraction of HSCs and multipotent progenitors have multilineage potential at the single-cell level. These new insights prompt an erythroid-focused model of hematopoietic differentiation., Graphical Abstract, Highlights • RBCs are the predominant cell type produced by multipotent hematopoietic progenitors • All cell types with myeloid potential also produced RBCs and platelets in vivo • Single HSCs and MPPF cells are capable of multilineage hematopoietic reconstitution • Erythroid cell production emerges as a default hematopoietic fate, In this article, Forsberg and colleagues use quantitative reconstitution assays to demonstrate that all myeloid-competent progenitor cells also contribute to erythroid and platelet production. Single-cell in vivo analyses showed that hematopoietic stem cells and multipotent progenitors are clonally multipotent. These results suggest that erythropoiesis is a default hematopoietic fate and challenges current views on hematopoietic lineage potential and fate decisions.

Despite success in hematologic malignancies, chimeric antigen receptor (CAR) T cells have been less effective in solid tumors, in part because of the heterogeneous expression of the CAR-specific target antigen (1° Ag) found within the tumor mass. In combination with a CAR, bispecific T cell engagers (BiTEs) can target additional tumor antigens (2° Ag) while engaging CD3 signaling molecules on T cells, providing a unique way to address tumor heterogeneity, mitigate antigen escape and further potentiate durable effector function. However, in generating off-the-shelf universal CAR-T and NK cells, a combination strategy is not feasible as neither modified cell product is compatible to specifically engage with a BiTE. In developing allogeneic CAR-T cells, the T cell receptor (TCR) surface expression must be eliminated to prevent graft versus host disease, but the absence of surface TCR expression leads to loss of surface CD3 expression and BiTE compatibility. Similarly, NK cells naturally lack TCR expression and have no surface CD3 molecules for BiTE engagement. Here we discuss the development of a novel chimeric CD3ε fusion receptor (CD3-CFR) with a modified transmembrane and endodomain enabling surface expression in TCR-less T or NK cells, allowing for a novel combinatorial solution between universal CAR-T or NK cells with BiTEs in an allogeneic setting. CD3-CFR signal transduction was initially investigated in TRAC knockout NFAT reporter (T-KO) Jurkat cells engineered with CD3-CFR and co-cultured with EpCAM+ target cells. When soluble EpCAM-BiTE was added to the co-culture, the observed 6.9-fold increase in NFAT activity indicated successful engagement associated with the interaction of CD3-CFR, target cells and the BiTE. CD3-CFR T-KO Jurkat cells were further modified to secrete the EpCAM-BiTE and showed increased NFAT activity, demonstrating the feasibility of self-secreting BiTE-mediated targeting through CD3-CFR signaling. We next engineered CD3-CFR expressing iPSC-derived CAR-T (iT) cells to show preserved T cell differentiation kinetics and maintained CAR activity against 1° Ag positive targets (>90% cytolysis). Importantly, while CAR-iT cells showed only background lysis of 1° Ag negative tumor cells, CD3-CFR+ CAR-iT cells were able to lyse ~99% of 1° Ag negative tumor cells with the addition of soluble EpCAM-BiTE targeting the 2° Ag. When CD3-CFR+ CAR-iT cells were further engineered to secrete the EpCAM-BiTE, CD3-CFR+ BiTE+ CAR-iT cells demonstrated superior killing of heterogenous tumors compared to CD3-CFR+ BiTE- CAR-iT cells (70% vs. 5% cytolysis). Taken together these data demonstrate for the first time that CD3 expression and BiTE engagement can be combined in a universal iPSC-derived CAR-iT cell product to overcome antigen heterogeneity and enhance efficacy in solid tumor settings. Citation Format: Eigen Peralta, Dan Lu, Mark Landon, Hui-Yi Chu, Amit Mehta, Philip Chu, Alec Witty, Tom Lee, Bahram Valamehr. Chimeric CD3 fusion receptors expressed on iPSC-derived universal TCR-less CAR-T and -NK cells synergize with bispecific engagers to enhance antitumor activity and limit antigen escape [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5513.

Immune cell subtype population frequencies can have a large effect on the efficacy of T cell therapies. Current methods, like flow cytometry, have specific sample requirements, high sample input, are low throughput, and are difficult to standardize, all of which are detrimental to characterization of cell therapy products during their development and manufacturing. The assays described herein accurately identify and quantify immune cell types in a heterogeneous mixture of cells using isolated genomic DNA (gDNA). DNA methylation patterns are revealed through bisulfite conversion, a process in which unmethylated cytosines are converted to uracils. Unmethylated DNA regions are detected through qPCR amplification using primers targeting converted areas. One unique locus per assay is measured and serves as an accurate identifier for a specific cell type. The assays are robust and identify CD8+, regulatory, and Th17 T cells in a high throughput manner. These optimized assays can potentially be used for in-process and product release testing for cell therapy process.

T cells require a combination of signals to properly activate and exhibit their full potential; signal 1 from the T cell receptor (TCR), signal 2 from costimulatory receptors and signal 3 from cytokine receptors can shape the function of T cells. Second generation chimeric antigen receptor (CAR) T-cells targeting CD19 have shown success in hematologic malignancies, in part because signals 1 and 2 are provided by the CAR endodomain. As previously reported, we have developed an off-the-shelf iPSC-derived CAR19 T (CAR19-iT) cell product where the CAR is inserted into the T cell receptor alpha constant (TRAC) locus for enhanced efficacy and disruption of surface TCR expression. While the TCR-less CAR19-iT cells are ideal for allogeneic CAR-T cell therapies, as a consequence, they lack CD3 surface expression and the means by which to mediate CD3 signaling. To further improve CAR-iT cell performance and facilitate combinatorial strategies with T cell engagers such as bispecific T cell engagers (BiTEs), we designed chimeric fusion receptors (CFRs) comprising an ectodomain of CD3 or CD28, a transmembrane domain of CD28, and various endodomains that provide combinations of signals 1, 2, and 3. In addition to providing co-stimulatory signaling, the CFR designs can be controlled via agonistic antibodies or BiTEs to initiate signal transduction for precision control of the cell product. Furthermore, a series of CFRs with various CD3 endodomains were also developed to augment costimulatory signaling and the ER retention and endocytosis motifs found in CD3 endodomains were mutated to prevent downregulation of the CFRs from the cell surface of TCR-less cells. Selected CFRs were engineered into TRAC knockout Jurkat cells expressing a luciferase based NFAT reporter gene. After 48 hours, surface expression of the CFRs was confirmed by flow cytometry. Signal transduction via the CFRs was evaluated following exposure to agonistic antibodies or BiTEs. Measurement of the luciferase activity showed comparable levels to activated wildtype reporter cells. After construct validation, the CFRs were introduced into TCR-less CAR19-iT cells and co-cultured overnight with Nalm6 target cells in the presence of agonistic antibodies. Compared to control CAR19-iT cells, the armed CFR-CAR19 iT cells demonstrated superior killing of target cells. These results support the notion that enhancing signal 1 can increase CAR-T cell efficacy with current studies underway to investigate CFRs with endodomains that provide additional signal 2 or 3. We expect these constructs to further enhance CAR-iT cell persistence and engagement with BiTEs for activation and multi-antigen targeting. Collectively, the data suggest that TRC-less CAR-iT cells can be further armed with CFRs containing appropriate endodomains to provide superior antitumor function in a regulated manner. Citation Format: Eigen Peralta, Dan Lu, Mark Landon, Helen Chu, Shohreh Sikaroodi, Emily Carron, Natalie Navarrete, Alec Witty, Tom Lee, Anhco Nguyen, Bahram Valamehr. Chimeric fusion receptors provide a CD3-mediated activation signal to off-the-shelf iPSC-derived TCR-less CAR-T cells for enhanced efficacy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 332.

T cell-based immunotherapies offer a potent and specific solution to grievous diseases such as cancer and viral infections. Expansion and re-infusion of antigen specific T cells has seen success in the treatment of metastatic melanoma and prevention of viral infections such as CMV and EBV. Additionally, engineered T cell therapies, especially chimeric antigen receptor (CAR) T cells, have been met with great clinical success and spurred innovation within the field of cellular immunotherapies. While promising, these therapies rely on sourcing T cells from donor derived blood. This process is invasive to the patient, especially when multiple or prolonged blood draws are required to obtain sufficient T cell numbers. Additionally, T cells sourced from diseased patients are often exhausted and display significantly perturbed growth and dysfunctional responses to antigen stimulation. An appealing alternative is somatic reprogramming of T cells to generate induced pluripotent stem cells (iPSCs) and differentiated into rejuvenated T cells that retain the original TCR, particularly with tumor infiltrating lymphocytes or viral specific T cells. We had previously reported a Sendai-virus based CytoTune 2.1 Reprogramming specifically designed for translational and clinical research. In this study we examine the effect of various cellular phenotypes and optimize methods for consistent reprogramming of multiple donor T cell reprogramming. Additionally, pan CD3+ cells isolated using different methods and further enriched for CD4+ and CD8+ were successfully reprogrammed. The resulting iPSCs were extensively characterized to ensure that pluripotency was achieved and that TCR sequences were preserved. This approach of reprogramming T cells and specific cell subtypes to derive iPSC with a known TCR sequence enables their use for downstream application as a renewable source of T cells with known antigen specificity. Citation Format: Mark Landon, Chad C. MacArthur, Melissa E. Hernandez, Uma Lakshmipathy, Jerry Guzman. Establishing consistent methods for the generation of donor-derived iPSCs from T cell subsets [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 488.

Background & Aim The clinical success of approved chimeric antigen receptor T-Cell (CAR-T) therapy to treat B-cell leukemias has generated interest to apply similar approach to other forms of cancer. However, processing patient T cells that are often exhausted and dysfunctional poses a challenge. This and the steep cost associated with autologous therapies has led to exploration of allogeneic approaches using T cells from healthy donors. An appealing alternate source are induced pluripotent stem cells (iPSCs) since they can be expanded indefinitely and differentiated into healthy T cells. Reprogramming T cells to generate T-iPSC is especially beneficial when working with antigen specific T cells, such as tumor infiltrating lymphocytes or viral specific T cells, where preserving antigen recognition is of importance. Additionally, T-cell derived iPSC (T-iPSC) with an unique TCR serves as an intrinsic barcode enabling in vivo tracking that can be a beneficial for any iPSC-based therapy. Methods, Results & Conclusion We had previously reported a Sendai-virus based reprogramming specifically designed for translational and clinical applications. Here, we studied the effect of diverse cellular phenotypes in different donor T cells on reprogramming. A robust method of reprogramming was established for CD3+ cells isolated using different methods and enriched for CD4+ and CD8+ T cells. Resulting iPSC were analyzed using NGS-based immune repertoire to generate high-throughput sequencing data to examine the diversity of TCR. Comprehensively characterized T-iPSC cell banks offer a renewable cell source that can be differentiated into cell types of choice for the development of translational therapies.

Background & Aim Manufacturing cell therapy products often rely on laborious cell isolation techniques that result in contaminating byproducts. In order to ensure product quality and consistency, thorough tests are performed throughout the manufacturing process and for product release. Characterization of a biological product includes determination of biological activity, identity, purity and impurities. Identity and purity assays commonly rely on cell surface markers or secreted molecules that are measured using flow cytometry. A critical challenge with flow cytometry is the need for live samples and lack of standardization due to biological, technical and operational variability. An appealing alternative that overcomes these constraints is the PureQuant assay, a cell type-specific, epigenetic qPCR based method that is sensitive, consistent and standardized. Methods, Results & Conclusion PureQuant assays were evaluated in CAR-T cells using assays specific for T-cell specific CD3, CD4, CD8, and contaminating B cells and monocytes. The individual assays each comprise of a reference to check assay performance, calibrator that corrects for bisulfite conversion efficiencies and standards to deduce the exact copy number. The panel of five assays was used to determine the purity of multiple healthy donor-derived CAR-T cells generated using a second generation (CD3ζ and 4-1BB) anti-CD19 CAR lentivirus. Sensitivity to detect contaminating cells was further established by spike-in experiments with addition of purified B-cell and monocytes to CAR-T cells. These studies demonstrate that the combination of accuracy, low sample requirement and flexibility provides an ideal measurement system for confirmation of identity and purity critical for therapeutic applications.

Significant progress has been made in harnessing the power of immune system, in particular, T cells, to treat certain kinds of cancers. One of the key challenges in developing immune cells as therapeutic agents is the accurate estimation of their identity and purity. Current methods used for the characterization of immune cell types rely on flow cytometry. Flow cytometry can accurately estimate CD8+ T lymphocytes and other surface markers, but the method is challenging to implement in a GMP manufacturing environment posing logistical challenges such as requirement for live cells, variability leading to difficult in standardizing and high throughput. In addition, cytometric methods are not accurate for specific intracellular targets that positively identify Regulatory T (Treg) cells and T Helper 17 (Th17) cells. Therefore, there is an emerging need for alternative assay methods. Methylation state is known to be unique for specific cell types and can thus be used as an identifier in heterogeneous population of cells. Exploiting differences in cell type-specific methylation signatures, we developed assay kits that quantify the percentage of Treg and Th17 by detecting methylation status of FoxP3 and IL17A via qPCR of bisulfite converted genomic DNA. In contrast to flow analysis, sample requirement is minimal and the assay works well with fresh/frozen cells or genomic DNA. This assay has been implemented to accurately identify and estimate different T cell population in Chimeric Antigen Receptor (CAR)-modified T cells. The combination of accuracy, low sample requirement and flexibility provides an ideal measurement system for confirmation of identify and purity of T cell types critical for therapeutic applications.

Background & Aim Induced pluripotent stem cell (iPSC) research is rapidly moving towards translational and clinical applications.These applications require robust and consistent workflows that utilize high quality reagents that are xeno-free.When commonly used animal-origin components are replaced with xeno-free alternatives, performance often suffers,thereby necessitating thorough qualification of reagents and the process.Methods that enhance consistency will minimize extra effort and costs associated with clones that fail to expand,or do not meet quality standards for downstream use. Methods, Results & Conclusion Previously,we reported the first off-the-shelf reprogramming kit specifically designed for clinical and translational research.To streamline iPSC generation and ensure the consistent creation of high quality iPSCs,xeno-free workflows for T cells were optimized to minimize the effect of donor to donor variability.Upfront phenotyping of the T-cells prior to reprogramming was performed,along with optimization of a combination of conditions including hypoxia,matrix,and seeding density.These optimizations were shown to offer consistent iPSC generation from difficult to reprogram donor cells,albeit with varying efficiencies.To confirm the parental cell type,T-cell derived iPSC clones were analyzed for immune repertoire,leveraging a next-generation sequencing based assay.The resulting iPSC lines were also confirmed to be foot-print free,authenticated to be derived from the corresponding donor cells,HLA profiled,and were further subjected to comprehensive characterization methods to assess the quality and safety profile.Pluripotency and differentiation potential of iPSC clones was confirmed using ScoreCard,a focused qPCR panel,and PluriTest,an array-based global gene expression platform.Clones qualified to be pluripotent and possess trilineage differentiation potential were thoroughly investigated for genomic stability.Each of the clones tested showed a normal karyotype using both traditional G-banding as well as array-based methods KaryoStat and KaryoStat HD.Further assessment of oncogenic hotspot mutations that are known to occur at a higher frequency in ESC and iPSC,including TP53,indicated the absence of oncogenic structural variants in all the clones tested.The combination of qualified reagents and defined xeno-free workflows, in combination with comprehensive and predictive characterization assays aids in easy transition of early investigational work towards translational and clinical research.

Selective labeling of specific cell types by expression of green fluorescent protein (GFP) within the hematopoietic system would have great utility in identifying, localizing, and tracking different cell populations in flow cytometry, microscopy, lineage tracing, and transplantation assays. In this report, we describe the generation and characterization of a new transgenic mouse line with specific GFP labeling of all nucleated hematopoietic cells, as well as platelets. This new “Vav-GFP” mouse line labels the vast majority of hematopoietic cells with GFP during both embryonic development and adulthood, with particularly high expression in hematopoietic stem and progenitor cells. With the exception of transient labeling of fetal endothelial cells, GFP expression is highly selective for hematopoietic cells, and persists in donor-derived progeny after transplantation of hematopoietic progenitor cells. Finally, we also demonstrate that the loxP-flanked reporter allows for specific GFP labeling of different hematopoietic cell subsets when crossed to various Cre reporter lines. Notably, by crossing Vav-GFP mice to Flk2-Cre mice, we obtained robust and highly selective GFP expression in HSCs. Together, these data describe a new mouse model capable of directing GFP labeling exclusively of hematopoietic cells or exclusively of HSCs.

OBJECTIVE: To compare the rates and perinatal outcome in women who experienced preeclampsia in a previous pregnancy to those in women who developed preeclampsia as nulliparas. STUDY DESIGN: This is a secondary analysis of data from 2 separate multi-center trials of aspirin for prevention of preeclampsia. Women who had preeclampsia in a previous pregnancy (n = 598) were compared with nulliparous women (n = 2934). Outcome variables were rates of preeclampsia, preterm delivery at

The National Ignition Facility (NIF) is under construction at Lawrence Livermore National Laboratory for the DOE Stockpile Stewardship Program. It will be used for experiments for inertial confinement fusion ignition, high energy density science, and basic science. Many interface issues confront the experimentalist who wishes to design, fabricate, and install diagnostics, and to help this process, a set of standards and guideline documents is being prepared. Compliance with these will be part of a formal diagnostic design review process. In this article we provide a short description of each, with reference to more complete documentation. The complete documentation will also be available through the NIF Diagnostics web page. Target area interface issues are grouped into three categories. First are the layout and utility interface issues which include the safety analysis report, target area facility layout; target chamber port locations; diagnostic interferences and envelopes; utilities and cable tray dist...

Objective: This study was undertaken to compare rates and severity of gestational hypertension and preeclampsia, as well as perinatal outcomes when these complications develop, between women with twin gestations and those with singleton gestations. Study Design: This was a secondary analysis of prospective data from women with twin (n = 684) and singleton (n = 2946) gestations enrolled in two separate multicenter trials of low-dose aspirin for prevention of preeclampsia. End points were rates of gestational hypertension, rates of preeclampsia, and perinatal outcomes among women with hypertensive disorders. Results: Women with twin gestations had higher rates of gestational hypertension (relative risk, 2.04; 95% confidence interval, 1.60-2.59) and preeclampsia (relative risk, 2.62; 95% confidence interval, 2.03-3.38). In addition, women with gestational hypertension during twin gestations had higher rates of preterm delivery at both P P P = .04). Moreover, when outcomes associated with preeclampsia were compared, women with twin gestations had significantly higher rates of preterm delivery at P P P = .07). In contrast, among women with twin pregnancies, those who remained normotensive had more adverse neonatal outcomes than did those in whom hypertensive complications developed. Conclusions: Rates for both gestational hypertension and preeclampsia are significantly higher among women with twin gestations than among those with singleton gestations. Moreover, women with twin pregnancies and hypertensive complications have higher rates of adverse neonatal outcomes than do those with singleton pregnancies. (Am J Obstet Gynecol 2000;182:938-42.)

Objective: The aim of this study was to determine whether epidural anesthesia during labor increased the frequencies of cesarean delivery, pulmonary edema, and renal failure among women with severe hypertensive disease. Study Design: We performed a secondary retrospective analysis of a subgroup population within a multicenter double-blind trial of low-dose aspirin therapy for women at high risk for development of preeclampsia. Subjects in whom severe hypertensive disease developed were selected. The primary outcomes were the overall frequencies of cesarean delivery among women with severe hypertensive disease who had labor with and without epidural anesthesia. Other maternal and neonatal outcomes were also compared between women who did and did not receive epidural anesthesia. Results: Among the women with severe hypertensive disease (n = 444) 327 had labor. Among the women with severe disease who had labor there was no difference in either the overall cesarean delivery rate (32.1% vs 28.0%; P = .44) or the rate of cesarean delivery for fetal distress or failure to progress (27.8% vs 22.0%; P = .26) between women who did and did not receive epidural analgesia. Women with chronic hypertension were more likely to have a cesarean delivery overall if they received epidural anesthesia, but there was otherwise no difference in the frequencies of cesarean delivery for these indications between women with and without epidural anesthesia within each of the high-risk groups. Pulmonary edema was rare and acute renal failure did not develop in any women. Conclusion: Epidural anesthesia use did not increase the frequencies of cesarean delivery, pulmonary edema, and renal failure among women with severe hypertensive disease. (Am J Obstet Gynecol 1999;181:1096-1101.)

Background Whether low-dose aspirin prevents preeclampsia is unclear. It is not recommended as prophylaxis in women at low risk for preeclampsia but may reduce the incidence of the disease in women at high risk. Methods We conducted a double-blind, randomized, placebo-controlled trial in four groups of pregnant women at high risk for preeclampsia, including 471 women with pregestational insulin-treated diabetes mellitus, 774 women with chronic hypertension, 688 women with multifetal gestations, and 606 women who had had preeclampsia during a previous pregnancy. The women were enrolled between gestational weeks 13 and 26 and received either 60 mg of aspirin or placebo daily. Results Outcome data were obtained on all but 36 of the 2539 women who entered the study. The incidence of preeclampsia was similar in the 1254 women in the aspirin group and the 1249 women in the placebo group (aspirin, 18 percent; placebo, 20 percent; P = 0.23). The incidences in the aspirin and placebo groups for each of the four hi...

Recently discovered Haynesville gas-shale trends have transformed the regional and global outlook for natural gas supply, but offer unique challenges to the operator and service company during mud removal, cementing, and completion operations. To counter these challenges, recent advances include improved drilling, centralization, mud-removal, cementing best practices and implementing a broad particle size distribution-engineered (PSDE) cement system for use in high temperature horizontal intervals reaching across high pressure, high temperature (HPHT) gas-shale trends. For PSDE cement systems, rheological properties are based on inter-particulate interactions to achieve the desired viscosity and not based on polymeric extenders/antisettling additives. Since PSDE fluids are not dependent on polymeric thermal thinning behavior, they demonstrate consistent rheological properties over a wide temperature range and are more suitable for placement in narrow annuli.In this paper, Haynesville cement placement and extensive laboratory testing best practices will be discussed. Also, a case study will be presented that describes a typical and successful placement of PSDE cement fluid in the Haynesville shale at bottom hole circulating temperatures (BHCT) up to 182 °C [360 °F] and bottom hole pressures (BHP) up to 82.7 MPa [12,000 psi]. After successful job completion and time allowed for the cement to properly set, an annular seal pressure test was successfully completed, with minimal pressure bleed-off.Since introduction in 2009, over 390 production jobs have been successfully cemented using the PSDE cement technology, with 99.5% placement success rate. Acquired well head pressures (WHP) were less than or equal to predicted WHP for most production jobs. PSDE Cement Technology has become a proven approach for cementing high-temperature, horizontal tight-gas shale environments in relatively narrow annuli where fluid stability and zonal isolation are needed during placement and subsequent hydraulic fracturing treatments. This approach has been applied to Haynesville and Eagleford shale horizontal reach production wells and is being investigated for use in other high temperature, high pressure applications.

To determine whether asthma-specific quality of life during pregnancy is related to asthma exacerbations and to perinatal outcomes.This was a secondary analysis of data from a randomized controlled trial of inhaled beclomethasone versus theophylline in the treatment of moderate asthma during pregnancy. The Juniper Asthma Quality of Life Questionnaire (AQLQ) was administered to patients at enrollment. Exacerbations were defined as asthma symptoms requiring a hospitalization, unscheduled medical visit, or oral corticosteroid course.Quality of life assessments were provided by 310 of the 385 participants who completed the study. There was more than a 25% decrease in the odds of a subsequent asthma exacerbation for every 1-point increase in AQLQ score for the overall score (odds ratio [OR] 0.73, 95% confidence interval [CI] 0.55-0.96), emotion domain (OR 0.72, 95% CI 0.59-0.88), and symptoms domain (OR 0.73, 95% CI 0.57-0.94). These relationships were not significantly influenced by initial symptom frequency or forced expiratory volume in 1 s (FEV(1)). No significant relationships were demonstrated between enrollment AQLQ scores and preeclampsia, preterm birth, low birth weight, or small for gestational age.Asthma-specific quality of life in early pregnancy is related to subsequent asthma morbidity during pregnancy but not to perinatal outcomes.

The current literature emphasizes increased risk of adverse outcomes in the presence of proteinuria and hypertension. The objective of this study was to compare the frequency of adverse fetal outcomes in women who developed hypertensive disorders with or without proteinuria.The study design was a secondary analysis of data from women who had preeclampsia in a previous pregnancy (n = 598) who were enrolled in a multicenter trial of aspirin for the prevention of preeclampsia. The women had no history of chronic hypertension or renal disease and were normotensive at study inclusion. The maternal and perinatal outcome variables assessed were preterm delivery at37 and35 weeks of gestation, rate of small-for-gestational-age infants, and abruptio placenta. Data were analyzed by using the chi-square test, and women who remained normotensive or who had mild gestational hypertension were considered as a single group because they had similar outcomes.As compared to mild preeclampsia, women who developed severe gestational hypertension (without proteinuria) had higher rates of both preterm delivery at37 weeks of gestation and small-for-gestational-age infants. In addition, when compared to women with mild preeclampsia, for women with severe gestational hypertension, gestational age and birth weight were significantly lower at delivery (P.003 for both age and birth weight). Moreover, women who developed severe gestational hypertension had higher rates of preterm delivery at37 weeks of gestation (54.2% vs 17.8%, P =.001) and at35 weeks of gestation (25.0% vs 8.4%, P =.0161), and delivery of small-for-gestational-age infants (20.8% vs 6.5%, P =.024) when compared to women who remained normotensive or those who developed mild gestational hypertension. There were no statistically significant differences in perinatal outcomes between the normotensive/mild gestational hypertension and the mild preeclampsia groups. Overall, women who had severe gestational hypertension had increased rates of preterm delivery and delivery of small-for-gestational-age infants than women with mild gestational hypertension or mild preeclampsia. In the presence of severe hypertension, proteinuria did not increase the rates of preterm delivery or delivery of small-for-gestational-age infants.In women who have gestational hypertension or preeclampsia, increased rates of preterm delivery and delivery of small-for-gestational-age infants are present only in those with severe hypertension. In these women, the presence of proteinuria does not influence perinatal outcome.

We have developed a Ti:sapphire/Nd:glass laser system which produces > 1.25 PW peak power. An irradiance of 1020 - 1021 W/cm2 is achieved utilizing an on-axis parabolic mirror, with adaptive optic wavefront correction. Experimental results will be described.© (1999) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Although there have been several anecdotal reports of the use of buccal pulse oximeter monitoring (Spo2) when digital Spo2 monitoring cannot be used, there have been no objective evaluations of the accuracy of buccal Spo2 monitoring. The purpose of this study was to systematically compare buccal Spo2 monitoring to both digital Spo2 and arterial O2 saturation monitoring (Sao2) in both generally anesthetized patients in the operating room (n = 31) and critically ill patients in the intensive care unit (n = 23). Buccal Spo2 probes were prepared by taping a malleable metal bar securely over the back of a Nellcor Oxisensor D-25 probe and bending the metal bar and buccal probe firmly around the corner of the patient's mouth. All buccal and finger Spo2 and Sao2 measurements were made simultaneously during hemodynamic stability. We found that buccal Spo2 was higher than finger Spo2 and agreed more closely with Sao2 for both patient groups (98.1% +/- 2.6%, 96.8% +/- 3.5%, 98.5% +/- 2.5%, respectively [mean +/- SD]). The operating room patients had higher buccal and finger Spo2 and Sao2 (99.3% +/- 1.5%, 98.9% +/- 1.4%, 99.5% +/- 0.7%, respectively) than the intensive care unit patients (96.4% +/- 2.9%, 94.1% +/- 3.5%, 96.6% +/- 3.5%, respectively). Although buccal Spo2 monitoring has several disadvantages (i.e., the probe requires preparation, can be more difficult to place, may be less readily accepted in awake patients, and is often mechanically dislodged during airway maneuvers), we conclude that buccal Spo2 monitoring is a more than adequate oximetry alternative when digital Spo2 monitoring is not an option (digits are unavailable or available digits are mechanically interfered with).