Cleaning verification by air/water impingement

This paper will discuss how the Kennedy Space Center intends to perform precision cleaning verification by Air/Water Impingement in lieu of chlorofluorocarbon-113 gravimetric nonvolatile residue analysis (NVR). Test results will be given that demonstrate the effectiveness of the Air/Water system. A brief discussion of the Total Carbon method via the use of a high temperature combustion analyzer will also be given. The necessary equipment for impingement will be shown along with other possible applications of this technology

Effects of Anacetrapib in Patients with Atherosclerotic Vascular Disease

BACKGROUND: Patients with atherosclerotic vascular disease remain at high risk for cardiovascular events despite effective statin-based treatment of low-density lipoprotein (LDL) cholesterol levels. The inhibition of cholesteryl ester transfer protein (CETP) by anacetrapib reduces LDL cholesterol levels and increases high-density lipoprotein (HDL) cholesterol levels. However, trials of other CETP inhibitors have shown neutral or adverse effects on cardiovascular outcomes. METHODS: We conducted a randomized, double-blind, placebo-controlled trial involving 30,449 adults with atherosclerotic vascular disease who were receiving intensive atorvastatin therapy and who had a mean LDL cholesterol level of 61 mg per deciliter (1.58 mmol per liter), a mean non-HDL cholesterol level of 92 mg per deciliter (2.38 mmol per liter), and a mean HDL cholesterol level of 40 mg per deciliter (1.03 mmol per liter). The patients were assigned to receive either 100 mg of anacetrapib once daily (15,225 patients) or matching placebo (15,224 patients). The primary outcome was the first major coronary event, a composite of coronary death, myocardial infarction, or coronary revascularization. RESULTS: During the median follow-up period of 4.1 years, the primary outcome occurred in significantly fewer patients in the anacetrapib group than in the placebo group (1640 of 15,225 patients [10.8%] vs. 1803 of 15,224 patients [11.8%]; rate ratio, 0.91; 95% confidence interval, 0.85 to 0.97; P=0.004). The relative difference in risk was similar across multiple prespecified subgroups. At the trial midpoint, the mean level of HDL cholesterol was higher by 43 mg per deciliter (1.12 mmol per liter) in the anacetrapib group than in the placebo group (a relative difference of 104%), and the mean level of non-HDL cholesterol was lower by 17 mg per deciliter (0.44 mmol per liter), a relative difference of -18%. There were no significant between-group differences in the risk of death, cancer, or other serious adverse events. CONCLUSIONS: Among patients with atherosclerotic vascular disease who were receiving intensive statin therapy, the use of anacetrapib resulted in a lower incidence of major coronary events than the use of placebo. (Funded by Merck and others; Current Controlled Trials number, ISRCTN48678192 ; ClinicalTrials.gov number, NCT01252953 ; and EudraCT number, 2010-023467-18 .)

Maternal SARS-CoV-2 infection in pregnancy disrupts gene expression in Hofbauer cells with limited impact on cytotrophoblasts.

BackgroundHofbauer cells (HBCs) and cytotrophoblasts (CTBs) are major cell populations in placenta. The indirect impact of maternal SARS-CoV-2 disease on these cells that are not directly infected has not been extensively studied. Herein, we profiled gene expression in HBCs and CTBs isolated from placentae of recovered pregnant subjects infected with SARS-CoV-2 during all trimesters of pregnancy, placentae from subjects with active infection, SARS-CoV-2 vaccinated subjects, and those who were unexposed to the virus.MethodsPlacentae were collected within 4 h post-delivery and membrane-free tissues were enzymatically digested for the isolation of HBCs and CTBs. RNA extracted from HBCs and CTBs were sequenced using 150bp paired-end reads. Differentially expressed genes (DEGs) were identified by DESeq2 package in R and enriched in GO Biological Processes, KEGG Pathway, Reactome Gene Sets, Hallmark Gene Sets, and Canonical Pathways. Protein-protein interactions among the DEGs were modelled using STRING and BioGrid.ResultsPregnant subjects (n = 30) were recruited and categorized into six groups: infected with SARS-CoV-2 in i) the first (1T, n = 4), ii) second (2T, n = 5), iii) third (3T, n = 5) trimester, iv) tested positive at delivery (Delivery, n = 5), v) never infected (Control, n = 6), and vi) fully mRNA-vaccinated by delivery (Vaccinated, n = 5). Compared to the Control group, gene expression analysis showed that HBCs from infected subjects had significantly altered gene expression profiles, with the 2T group having the highest number of DEGs (1,696), followed by 3T and 1T groups (1,656 and 958 DEGs, respectively). These DEGs were enriched for pathways involved in immune regulation for host defense, including production of cytokines, chemokines, antimicrobial proteins, ribosomal assembly, neutrophil degranulation inflammation, morphogenesis, and cell migration/adhesion. Protein-protein interaction analysis mapped these DEGs with oxidative phosphorylation, translation, extracellular matrix organization, and type I interferon signaling. Only 95, 23, and 8 DEGs were identified in CTBs of 1T, 2T, and 3T groups, respectively. Similarly, 11 and 3 DEGs were identified in CTBs and HBCs of vaccinated subjects, respectively. Reassuringly, mRNA vaccination did not induce an inflammatory response in placental cells.ConclusionsOur studies demonstrate a significant impact of indirect SARS-CoV-2 infection on gene expression of inner mesenchymal HBCs, with limited effect on lining CTB cells isolated from pregnant subjects infected and recovered from SARS-CoV-2. The pathways associated with these DEGs identify potential targets for therapeutic intervention

Gene enrichment of interferon signaling pathway between trimesters versus control group (A and B) and delivery versus control group (C and D) in HBCs.

A) Enriched pathways of DEGs in HBCs of Trimester groups compared to Control group. B) The top 6 upregulated genes in Trimester groups compared to Control group. C) Enriched pathways of DEGs in HBCs of Trimester group compared to Control group. D) The top 6 downregulated genes in Delivery group compared to Control group.</p

Changes in DEGs when grouping subjects infected in the first, second, and third trimester (Trimester group) compared to delivery, vaccinated and control groups.

Number of DEGs in CTBs (A) and in HBCs (B). Orange: upregulated; Green: downregulated. C) Overlapping DEGs between HBCs isolated from the Trimester Group and Delivery group. Enriched ontology clusters of upregulated (D) and downregulated genes (E) in HBCs. F) Protein-protein interaction network of the significantly enriched biological pathways between Trimester versus Control groups and Infected versus Control groups.</p

Gene expression profile of HBC isolated from subjects testing SARS-CoV-2 positive at delivery as compared to the control group.

Enriched ontology clusters of (A) upregulated and (B) downregulated genes in HBCs. C) Protein-protein interaction network analysis demonstrated that these potentially dysregulated genes are involved in oxidative phosphorylation, translation, and cell cycle processes.</p

Gene expression profiles in term HBC infected across three trimesters (1T, 2T, 3T).

A) Number of DEGs in each comparison of HBCs across different groups. Orange: upregulated; Green: downregulated. B) Overlapping DEGs (Up: upregulated genes; Down: downregulated genes) in HBCs from subjects infected in the first (1T), second (2T), and third (3T) trimesters. C) Enriched topology clusters of upregulated and downregulated genes.</p

Study overview.

A) Study design. Pregnant subjects (n = 30) recruited for this study were categorized into 6 groups based on the time of infection: i) Four participants were infected during the first trimester (Group 1T), ii) Five infected during the second trimester (Group 2T), iii) Five infected during the third trimester (Group 3T), iv) Five tested positive by PCR at delivery time (Group Delivery), v) Six never infected (Control group), and vi) Five fully mRNA-vaccinated by delivery (Vaccinated group). Placentae were collected within 4 h following delivery. Cytotrophoblasts (CTBs) and Hofbauer cells (HBCs) were isolated from placentae, subjected to RNA extraction and subsequent mRNA sequencing. Gene expression profiles in CTBs and HBCs were compared across six different groups. Illustrations of Fig 1A was created using BioRender. B) Breakdown alignment for all sequenced samples (n = 60). One Hofbauer cell (HBCs) sample, 152-V-HCA, did not pass QC and was removed from downstream analysis. Therefore, a total of 59 cell samples of HBCs (n = 29) and CTBs (n = 30) were included in the downstream analysis. C) Principal component analysis (PCA) plot after removing one HC sample that did not pass QC. The gene expression in HBCs and CTBs were visually separated.</p

Gene expression profiles in CTBs.

A) Number of differentially expressed genes (DEGs) in each comparison of CTBs across different groups. Orange = upregulated; Green = downregulated. (A) Enriched topology clusters, (B) and enriched biological pathways (C) of DEGs in Group 1T compared to the Control group. D) Normalized counts of six genes encoding CXCL1, CXCL9, CXCL10, CXCL11, IL-32, CLCF-1 across groups.</p

Comorbidities of pregnant subjects included in this study.

Comorbidities of pregnant subjects included in this study.</p