Public Confidence Laws Gone Awry: A Modern Circuit Split Reveals that Some Federal Courts Manipulate Standing Rules to Promulgate Severe First Amendment Restrictions on the Spouses and Children of Public Employees

Federal courts in the United States have consistently upheld the constitutional doctrine that [t]he essential rights of the First Amendment in some instances are subject to the elemental need for order without which the guarantees of civil rights to others would be a mockery. Given the central role of government workers in maintaining that order, the First Amendment rights of public employees have been particularly susceptible to restriction. For example, in 1940, Congress enacted the Hatch Act, which declared unlawful certain political activities of federal employees. Specifically, section nine of the Act prohibited officers and employees in the executive branch from taking any active part in political management or in political campaigns. The theory behind restrictions such as those found in the Hatch Act relied on the state\u27s compelling interest in encouraging government impartiality and the public perception of impartiality in government. These restrictions, labeled public confidence laws, prohibit not only financial political contributions, but also contributions of time and energy in an effort to influence an election. The idea that a politically active government staff threatens effective administration has filtered down to the state and local level since the passage of the Hatch Act. Currently, not only are federal employees subject to public confidence laws, but many state and even local employees have been prohibited from participating in politics. While the principles justifying public confidence laws may indeed have deep judicial and constitutional roots, these laws have expanded at an alarming rate. The level of restriction they currently impose cannot be supported by any constitutional theory

A Guatemalan Soycow Cooperative: Is the Whole Greater than the Sum of its Parts?

Teaching Notes available upon request: [email protected]; Author video:http://www.youtube.com/user/ifamr1?feature=mhum#p/u/8/FhUZu2lt6NsSoycow, cooperative, Guatemala, teaching case, Agribusiness, Q10,

Konstruksi Backbone Plasmid Single Target dan tRNA-gRNA (TGR) Sistem CRISPR-Cas9 untuk Tanaman

Plasmid construction is one of many important steps within genome editing study using CRISPR-Cas9. Variety of genes that were targeted across different organisms and research’s purposes make it necessary to acquire the backbone plasmid with multiplexing capability (tRNA-gRNA) and flexibility to change its target by modifying crRNA sequences. Moreover, the size of plasmid also affects transformation efficiency. Therefore, it becomes clear that backbone plasmid is needed to be constructed with such characteristics. Golden Gate Assembly method was chosen with a combination of PCR reaction (henceforth called Golden Gate – PCR method) to complete two overlapping objectives, mutagenesis of BsaI recognition sequences at pVS1 Ori and combining 8 separate fragments, both backbone and insert PCR products. The results show that the final plasmid has been successfully constructed with a final size of 14,728 bp. PCR verification, short to full-length plasmid, of fragments’ order showed that all parts are assembled in intended positions. Furthermore, the most efficient method for applying Golden Gate – PCR is by directly amplifying Golden Gate reaction mixture with flanking primers (Direct PCR from Golden Gate). While the PCR reaction following purification of the correct Golden Gate product (Golden Gate – Purification – PCR) also yields the desired products, it consumes two purification reactions for each assembly. Capability to assemble only small fragments is the only limitation for Golden Gate – PCR method

Conformation, protein recognition and repair of DNA interstrand and intrastrand cross-links of Antitumor trans-[PtCl(2)(NH(3))(thiazole)]

Replacement of one ammine in clinically ineffective trans-[PtCl(2)(NH(3))(2)] (transplatin) by a planar N-heterocycle, thiazole, results in significantly enhanced cytotoxicity. Unlike ‘classical’ cisplatin {cis-[PtCl(2)(NH(3))(2)]} or transplatin, modification of DNA by this prototypical cytotoxic transplatinum complex trans-[PtCl(2)(NH(3))(thiazole)] (trans-PtTz) leads to monofunctional and bifunctional intra or interstrand adducts in roughly equal proportions. DNA fragments containing site-specific bifunctional DNA adducts of trans-PtTz were prepared. The structural distortions induced in DNA by these adducts and their consequences for high-mobility group protein recognition, DNA polymerization and nucleotide excision repair were assessed in cell-free media by biochemical methods. Whereas monofunctional adducts of trans-PtTz behave similar to the major intrastrand adduct of cisplatin [J. Kasparkova, O. Novakova, N. Farrell and V. Brabec (2003) Biochemistry, 42, 792–800], bifunctional cross-links behave distinctly differently. The results suggest that the multiple DNA lesions available to trans-planaramine complexes may all contribute substantially to their cytotoxicity so that the overall drug cytotoxicity could be the sum of the contributions of each of these adducts. However, acquisition of drug resistance could be a relatively rare event, since it would have to entail resistance to or tolerance of multiple, structurally dissimilar DNA lesions

Differences in Breast Cancer Diagnosis and Treatment:Experiences of Insured and Uninsured Patients in a Safety Net Setting

To explore how well the safety net performs at eliminating differences in diagnosis and treatment of insured and uninsured women with breast cancer, we compared insured and uninsured women treated in a safety net setting. Controlling for socioeconomic characteristics, uninsured women are more likely to be diagnosed with advanced disease, requiring more extensive treatment relative to insured women, and also experience delays in initiating and completing treatment. The findings suggest that, despite the safety net system, uninsured women with breast cancer are likely to require more costly treatment and to have worse outcomes, relative to insured women with breast cancer.

Construction and properties of a mutant of herpes simplex virus type 1 with glycoprotein H coding sequences deleted

A mutant of herpes simplex virus type 1 (HSV-1) in which glycoprotein H (gH) coding sequences were deleted and replaced by the Escherichia coli lacZ gene under the control of the human cytomegalovirus IE-1 gene promoter was constructed. The mutant was propagated in Vero cells which contained multiple copies of the HSV-1 gH gene under the control of the HSV-1 gD promoter and which therefore provide gH in trans following HSV-1 infection. Phenotypically gH-negative virions were obtained by a single growth cycle in Vero cells. These virions were noninfectious, as judged by plaque assay and by expression of I-galactosidase following high-multiplicity infection, but partial recovery of infectivity was achieved by using the fusogenic agent polyethylene glycol. Adsorption of gH-negative virions to cells blocked the adsorption of superinfecting wild-type virus, a result in contrast to that obtained with gD-negative virions (D. C. Johnson and M. W. Ligas, J. Virol. 62:4605-4612, 1988). The simplest conclusion is that gH is required for membrane fusion but not for receptor binding, a conclusion consistent with the conservation of gH in all herpesviruses

Mitigation of Iron and Aluminum Powder Deflagrations via Active Explosion Suppression in a 1 m3 Sphere Vessel

Combustible metal dust explosions continue to present a significant threat to metal handling and refining industries. Addition of noncombustible inert material to combustible dust mixtures, through either premixing or high-rate injection as the incipient flame front begins to develop, is common practice for preventative inhibition or explosion protection via active suppression, respectively. Metal dusts demonstrate an extremely reactive explosion risk due to amplified heat of combustion, burning temperature, flame speed, explosibility parameters (KSt and Pmax), and ignition sensitivity. Inhibition efficiency of suppressant agents used for active mitigation is shown to be reliant on fuel explosibility, discrete burning mechanism, and combustion temperature range and thus may be increasingly variable depending on the fuel in question. For this reason, mitigation of metal powder deflagrations at moderate total suppressed pressures (relative to the overall strength of the enclosure) and at low agent concentrations remains challenging. This paper reviews recent metal dust suppression testing in a Fike Corporation’s 1 m3 sphere combustion chamber and evaluates the efficacy of multiple suppression agents (sodium bicarbonate [SBC], sodium chloride [Met-L-X], and monoammonium phosphate [MAP]) for the mitigation of iron and aluminum powder deflagrations at suspended fuel concentrations of 2250 and 500 g/m3, respectively

Moving portraiture

"My finished work of art will take the form of a looped animation, created from a series of several hundred paintings. The piece is the video animation, not the paintings. The animation is comprised of several looped actions that are cut up and recombined to create new actions and movements that may not resemble the elements from which they were taken. My goal is to combine traditional portraiture with multimedia technology to create a hybrid work of art that is still understood as a portrait."--Abstract from author supplied metadata

Opioid use and harms associated with a sustained-release tapentadol formulation: a postmarketing study protocol

Introduction It has been argued that tapentadol may pharmacologically have lower abuse potential than other pharmaceutical opioids currently available. However, there has been no comprehensive triangulation of data regarding use and harms associated with this formulation. A sustained-release formulation (SRF) of tapentadol (Palexia) was released in Australia in 2011 and listed for public subsidy in 2013. We summarise here the methods of a postmarketing study which will measure postintroduction: (1) population level availability, (2) extramedical use and diversion, (3) attractiveness for extramedical use and (4) associated harms, of tapentadol compared against other pharmaceutical opioids. Methods and analysis We evaluated key sources on pharmaceutical use and harms in Australia. This review indicateddata from four sources that disaggregate pharmaceutical opioid formulations and capture tapentadol SRF could be triangulated. These data sources comprised: (1) national pharmaceutical opioid community sales data from 2011 to 2017, (2) national pharmaceutical opioid poisonings reported to Poison Information Centres (PICs) from 2011 to 2017, (3) number of vendors on online marketplaces listing pharmaceutical opioids for sale and (4) data on pharmaceutical opioid extramedical use, attractiveness and harms from interviews with people who regularly inject drugs in Australia. Ethics and dissemination Ethics approval is not required for use of pharmaceutical sales data. Ethics approval has been obtained for use of national pharmaceutical opioid poisonings reported to PICs (LNR/16/SCHN/44) and for use of online marketplace data and interview data from people who inject drugs (HC12086). Key findings will be published mid-2018 in a peer-reviewed academic journal, and presented at various conferences and professional meetings.This work was supported by investigator-initiated untied educational funding from Seqirus Pty Ltd (the marketer of tapentadol SRF in Australia) granted to AP, BL, MF, RC, and LD. BL, AP and LD are supported by NHMRC research fellowships (#1073858, #1109366 and #1041472). The National Drug and Alcohol Research Centre at UNSW Australia is supported by funding from the Australian Government under the Substance Misuse Prevention and Service Improvements Grant Fund

The CMV-encoded G protein-coupled receptors M33 and US28 play pleiotropic roles in immune evasion and alter host T cell responses.

INTRODUCTION: Human cytomegalovirus (HCMV) is a global health threat due to its ubiquity and lifelong persistence in infected people. During latency, host CD8+ T cell responses to HCMV continue to increase in a phenomenon known as memory inflation. We used murine CMV (MCMV) as a model for HCMV to characterize the memory inflation response to wild-type MCMV (KP) and a latency-defective mutant (ΔM33stop), which lacks M33, an MCMV chemokine receptor homolog. M33 is essential for normal reactivation from latency and this was leveraged to determine whether reactivation in vivo contributes to T cell memory inflation. METHODS: Mice were infected with wild-type or mutant MCMV and T cell responses were analyzed by flow cytometry at acute and latent time points. Ex vivo reactivation and cytotoxicity assays were carried out to further investigate immunity and virus replication. Quantitative reverse-transcriptase polymerase chain reaction (q-RTPCR) was used to examine gene expression during reactivation. MHC expression on infected cells was analyzed by flow cytometry. Finally, T cells were depleted from latently-infected B cell-deficient mice to examine the in vivo difference in reactivation between wild-type and ΔM33stop. RESULTS: We found that ΔM33stop triggers memory inflation specific for peptides derived from the immediate-early protein IE1 but not the early protein m164, in contrast to wild-type MCMV. During ex vivo reactivation, gene expression in DM33stop-infected lung tissues was delayed compared to wild-type virus. Normal gene expression was partially rescued by substitution of the HCMV US28 open reading frame in place of the M33 gene. In vivo depletion of T cells in immunoglobulin heavy chain-knockout mice resulted in reactivation of wild-type MCMV, but not ΔM33stop, confirming the role of M33 during reactivation from latency. Further, we found that M33 induces isotype-specific downregulation of MHC class I on the cell surface suggesting previously unappreciated roles in immune evasion. DISCUSSION: Our results indicate that M33 is more polyfunctional than previously appreciated. In addition to its role in reactivation, which had been previously described, we found that M33 alters viral gene expression, host T cell memory inflation, and MHC class I expression. US28 was able to partially complement most functions of M33, suggesting that its role in HCMV infection may be similarly pleotropic