Genomic structure and alternative splicing of murine R2B receptor protein tyrosine phosphatases (PTPκ, μ, ρ and PCP-2)

BACKGROUND: Four genes designated as PTPRK (PTPκ), PTPRL/U (PCP-2), PTPRM (PTPμ) and PTPRT (PTPρ) code for a subfamily (type R2B) of receptor protein tyrosine phosphatases (RPTPs) uniquely characterized by the presence of an N-terminal MAM domain. These transmembrane molecules have been implicated in homophilic cell adhesion. In the human, the PTPRK gene is located on chromosome 6, PTPRL/U on 1, PTPRM on 18 and PTPRT on 20. In the mouse, the four genes ptprk, ptprl, ptprm and ptprt are located in syntenic regions of chromosomes 10, 4, 17 and 2, respectively. RESULTS: The genomic organization of murine R2B RPTP genes is described. The four genes varied greatly in size ranging from ~64 kb to ~1 Mb, primarily due to proportional differences in intron lengths. Although there were also minor variations in exon length, the number of exons and the phases of exon/intron junctions were highly conserved. In situ hybridization with digoxigenin-labeled cRNA probes was used to localize each of the four R2B transcripts to specific cell types within the murine central nervous system. Phylogenetic analysis of complete sequences indicated that PTPρ and PTPμ were most closely related, followed by PTPκ. The most distant family member was PCP-2. Alignment of RPTP polypeptide sequences predicted putative alternatively spliced exons. PCR experiments revealed that five of these exons were alternatively spliced, and that each of the four phosphatases incorporated them differently. The greatest variability in genomic organization and the majority of alternatively spliced exons were observed in the juxtamembrane domain, a region critical for the regulation of signal transduction. CONCLUSIONS: Comparison of the four R2B RPTP genes revealed virtually identical principles of genomic organization, despite great disparities in gene size due to variations in intron length. Although subtle differences in exon length were also observed, it is likely that functional differences among these genes arise from the specific combinations of exons generated by alternative splicing

Systematic Screening of Drosophila Deficiency Mutations for Embryonic Phenotypes and Orphan Receptor Ligands

This paper defines a collection of Drosophila deletion mutations (deficiencies) that can be systematically screened for embryonic phenotypes, orphan receptor ligands, and genes affecting protein localization. It reports the results of deficiency screens we have conducted that have revealed new axon guidance phenotypes in the central nervous system and neuromuscular system and permitted a quantitative assessment of the number of potential genes involved in regulating guidance of specific motor axon branches. Deficiency “kits” that cover the genome with a minimum number of lines have been established to facilitate gene mapping. These kits cannot be systematically analyzed for phenotypes, however, since embryos homozygous for many deficiencies in these kits fail to develop due to the loss of key gene products encoded within the deficiency. To create new kits that can be screened for phenotype, we have examined the development of the nervous system in embryos homozygous for more than 700 distinct deficiency mutations. A kit of ∼400 deficiency lines for which homozygotes have a recognizable nervous system and intact body walls encompasses >80% of the genome. Here we show examples of screens of this kit for orphan receptor ligands and neuronal antigen expression. It can also be used to find genes involved in expression, patterning, and subcellular localization of any protein that can be visualized by antibody staining. A subset kit of 233 deficiency lines, for which homozygotes develop relatively normally to late stage 16, covers ∼50% of the genome. We have screened it for axon guidance phenotypes, and we present examples of new phenotypes we have identified. The subset kit can be used to screen for phenotypes affecting all embryonic organs. In the future, these deficiency kits will allow Drosophila researchers to rapidly and efficiently execute genome-wide anatomical screens that require examination of individual embryos at high magnification

Long-Term Durability and Cycling of Nanoporous Materials Based Impedance NO₂ Sensors

Nanoporous materials, including metal–organic frameworks (MOFs) and inorganic zeolites, are gaining attention as gas sensor materials due to their chemical selectivity and robustness. To advance industrial viability of these materials as sensors, long-term, variable environment testing is needed to evaluate their stability and continued chemical exposure response. Nanoporous materials-based direct electrical readout sensors were evaluated for 3 months under dry or humid conditions at 74 °C. The sensors were comprised of either Ni-MOF-74, Mg-MOF-74, or Ni-SSZ-13 zeolite. Additionally, we describe the development of multichambered sensor testing platforms that allows uninterrupted direct impedance monitoring of each sensor over long test periods. Results indicate relative stability in dry conditions for the sensors over time. In contrast, degradation of the active sensing material is evident in the humid environment. Collectively, these results demonstrate need for long-term testing of emerging nanoporous sensor materials under specific environmental conditions

The Use of Physiology-Based Pharmacokinetic and Pharmacodynamic Modeling in the Discovery of the Dual Orexin Receptor Antagonist ACT-541468

Stress-related psychiatric disorders across the life spa

Towards sustainable sanitation: Mitigating microbial health risks in the production of urine-derived fertilizers

To identify and address health and safety challenges, first, I conducted a screening of fecal pathogens in source-separated urine. The pathogen inactivation performance of two innovative fertilizer production technologies was then evaluated in laboratory and field conditions. An in-depth assessment of the main mechanisms leading to pathogen inactivation during fertilizer production suggested nuanced adjustments in the treatment to yield more hygienic outcomes. Finally, occupational exposure and risks of infection during fertilizer production were evaluated using innovative exposure analysis techniques and modeling. Quantitatively identifying risks in this way will allow development of strategies to promote innovation and overcome barriers to safe and sustainable sanitation