Long-Term Infection and Vertical Transmission of a Gammaretrovirus in a Foreign Host Species

Increasing evidence has indicated natural transspecies transmission of gammaretroviruses; however, viral-host interactions after initial xeno-exposure remain poorly understood. Potential association of xenotropic murine leukemia virus-related virus (XMRV) in patients with prostate cancer and chronic fatigue syndrome has attracted broad interests in this topic. Although recent studies have indicated that XMRV is unlikely a human pathogen, further understanding of XMRV xenoinfection would allow in vivo modeling of the initial steps of gammaretroviral interspecies transmission, evolution and dissemination in a new host population. In this study, we monitored the long-term consequences of XMRV infection and its possible vertical transmission in a permissive foreign host, wild-derived Mus pahari mice. One year post-infection, XMRV-infected mice showed no notable pathological changes, while proviral DNA was detected in three out of eight mice. XMRV-infected mice remained seropositive throughout the study although the levels of gp70 Env- and p30 capsid-specific antibodies gradually decreased. When vertical XMRV transmission was assessed, no viremia, humoral immune responses nor endogenization were observed in nine offspring from infected mothers, yet one offspring was found PCR-positive for XMRV-specific sequences. Amplified viral sequences from the offspring showed several mutations, including one amino acid deletion in the receptor binding domain of Env SU. Our results therefore demonstrate long-term asymptomatic infection, low incidence of vertical transmission and limited evolution of XMRV upon transspecies infection of a permissive new host, Mus pahari

Through the Alice Corp. Looking Glass: Using Pragmatic Arguments to Bring Predictability to Patent Law

Within the last four years, multiple institutional actors that have largely remained dormant in the scope of innovation policy and patents have undertaken a surprisingly interventionist reform of patent law. Congress, administrative agencies, and the Supreme Court have all recently shaped patent law to be more pragmatic and less overtly formalistic. Conversely, the Court of Appeals for the Federal Circuit has staunchly defended a formalist approach to patent law, advocating consistently for bright-line rules. The Federal Circuit has been charged with bringing predictability to patent law since its creation in 1982. The Federal Circuit attempts to bring predictability and uniformity through a largely rule-based formalist approach. Empirical evidence shows that the Federal Circuit\u27s rule formalism has actually decreased the predictability of patent law. The Supreme Court\u27s recent interventions into patent law show they are in favor of a more pragmatic approach, favoring policy considerations over the adoption of bright-line formalist rules. The history of the Federal Circuit shows it was designed to be an institutional body of expertise in patent law. This expertise was meant to serve two purposes, the first was to bring predictability and uniformity to patent law. The second was to be a body with not only the power to shape patent law, but the knowledge to shape it in accordance with the goals and policies inherent in the Patent Act. This comment explains how recent Supreme Court precedents regarding patent-eligible subject matter create a framework for inculcating pragmatic, policy intensive arguments into traditionally rigid, formalistic Federal Circuit jurisprudence. The framework becomes a vehicle for the Federal Circuit to shape patent law in alignment with the pragmatic and social considerations derived from innovation policy. This comment then illustrates how that framework can be applied to bring predictability to other key areas of uncertainty in patent law

Through the Alice Corp. Looking Glass: Using Pragmatic Arguments to Bring Predictability to Patent Law

Within the last four years, multiple institutional actors that have largely remained dormant in the scope of innovation policy and patents have undertaken a surprisingly interventionist reform of patent law. Congress, administrative agencies, and the Supreme Court have all recently shaped patent law to be more pragmatic and less overtly formalistic. Conversely, the Court of Appeals for the Federal Circuit has staunchly defended a formalist approach to patent law, advocating consistently for bright-line rules. The Federal Circuit has been charged with bringing predictability to patent law since its creation in 1982. The Federal Circuit attempts to bring predictability and uniformity through a largely rule-based formalist approach. Empirical evidence shows that the Federal Circuit\u27s rule formalism has actually decreased the predictability of patent law. The Supreme Court\u27s recent interventions into patent law show they are in favor of a more pragmatic approach, favoring policy considerations over the adoption of bright-line formalist rules. The history of the Federal Circuit shows it was designed to be an institutional body of expertise in patent law. This expertise was meant to serve two purposes, the first was to bring predictability and uniformity to patent law. The second was to be a body with not only the power to shape patent law, but the knowledge to shape it in accordance with the goals and policies inherent in the Patent Act. This comment explains how recent Supreme Court precedents regarding patent-eligible subject matter create a framework for inculcating pragmatic, policy intensive arguments into traditionally rigid, formalistic Federal Circuit jurisprudence. The framework becomes a vehicle for the Federal Circuit to shape patent law in alignment with the pragmatic and social considerations derived from innovation policy. This comment then illustrates how that framework can be applied to bring predictability to other key areas of uncertainty in patent law

Status of Biotic Integrity, Water Quality, and Physical Habitat in Wadeable East Texas Streams

United States Environmental Protection Agency, assistance agreement CR82548901Civil, Architectural, and Environmental Engineerin

Palmitate-Induced IRE1–XBP1–ZEB Signaling Represses Desmoplakin Expression and Promotes Cancer Cell Migration

Abstract Elevated uptake of saturated fatty acid palmitate is associated with metastatic progression of cancer cells; however, the precise signaling mechanism behind the phenomenon is unclear. The loss of cell adhesion proteins, such as desmoplakin (DSP), is a key driving event in the transformation of cancer cells to more aggressive phenotypes. Here, we investigated the mechanism by which palmitate induces the loss of DSP in liver and breast cancer cells. We propose that palmitate activates the IRE1–XBP1 branch of the endoplasmic reticulum (ER) stress pathway to upregulate the ZEB transcription factor, leading to transcriptional repression of DSP. Using liver and breast cancer cells treated with palmitate, we found loss of DSP leads to increased cell migration independent of E-cadherin. We report that the ZEB family of transcription factors function as direct transcriptional repressors of DSP. CRISPR-mediated knockdown of IRE1 confirmed that the transcription of ZEB, loss of DSP, and enhanced migration in the presence of palmitate is dependent on the IRE1–XBP1 pathway. In addition, by analyzing the somatic expression and copy number variation profiles of over 11,000 tumor samples, we corroborate our hypothesis and establish the clinical relevance of DSP loss via ZEB in human cancers. Implications: Provides mechanistic link on palmitate-induced activation of IRE1α to cancer cell migration. </jats:sec

Supplementary Figures 1-14 from Palmitate-Induced IRE1–XBP1–ZEB Signaling Represses Desmoplakin Expression and Promotes Cancer Cell Migration

S1A. Plasmids containing single guide RNAs targeted against the IRE1 locus. B. XBP1 splicing assay to assay IRE1 endoribonuclease activity in Hep3B and MDA MB 231 cells. S2. CRISPR clones obtained for MDA-MB-231 cells. S3. CRISPR clones obtained for Hep3B cells. S4. CRISPR clones obtained for HepG2 cells. S5. Representative brightfield images of Boyden's chamber migration assay. S6. Representative brightfield images of Boyden's chamber migration assay. S7. Representative brightfield images of Boyden's chamber migration assay. S8. Heatmap of standardized DSP, ZEB1 or ZEB2 expression in the TCGA PANCAN dataset. S9. Scatter plots showing the correlation between DSP methylation and the expression of DSP, ZEB1 or ZEB2 in the TCGA. S10A. Western blots showing expression levels of DSP, ZEB1, and GAPDH in MDA-MB-231 cells treated with scramble, no treatment, or siZEB1 at 100 nM (ORIGENE, cat # SR321982) for 48 hrs. The plots represent the quantification of the western blots. B-C. Barplots displaying relative gene expression levels measured using qRT-PCR of (B) ZEB1 and DSP (normalized to scramble) in MDA-MB-231 cells treated with siZEB1 at 100 nM 24 hrs, and (C) ZEB2 and DSP (normalized to scramble) in MDA-MB-231 cells treated with siZEB2 at 80 nM for 24 hrs. S11. A. Western blots showing expression levels of DSP, ZEB1, and GAPDH in Hep3B cells treated with scramble, no treatment, or siZEB1 at 100 nM (ORIGENE, cat # SR321982) for 48 hrs. The plots represent the quantification of the western blots. B-C. Barplots displaying relative gene expression levels measured using qRT-PCR of (B) ZEB1 and DSP (normalized to scramble) in Hep3B cells treated with siZEB1 at 100 nM 24 hrs, and (C) ZEB2 and DSP (normalized to scramble) in Hep3B cells treated with siZEB2 at 80 nM for 24 hrs. S12. Positive ChIP control for ZEB1 binding in Hep3B, MDA-MB-231 and HepG2 cells in the CDH1 promoter. S13. Relative ZEB1 protein expression in PA or BSA treated (48 hr) WT- or IRE1-KO MDA-MB-231 cells. S14. Relative ZEB1 protein expression in PA or BSA treated (72 hrs) WT- or IRE1-KO Hep3B cells.</p