The determinants of transmembrane protein insertion orientation at the endoplasmic reticulum have been investigated in Saccharomyces cerevisiae using variants of a Type III (naturally exofacial N terminus (N exo )) transmembrane fusion protein derived from the N terminus of Ste2p, the ␣-factor receptor. Small positive and negative charges adjacent to the transmembrane segment had equal and opposite effects on orientation, and this effect was independent of N-or C-terminal location, consistent with a purely electrostatic interaction with response mechanisms. A 3:1 bias toward N exo insertion, observed in the absence of a charge difference, was shown to reflect the N exo bias conferred by longer transmembrane segments. Orientation correlated best with total hydrophobicity rather than length, but it was also strongly affected by the distribution of hydrophobicity within the transmembrane segment. The most hydrophobic terminus was preferentially translocated. Insertion orientation thus depends on integration of responses to at least three parameters: charge difference across a transmembrane segment, its total hydrophobicity, and its hydrophobicity gradient. Relative signal strengths were estimated, and consequences for topology prediction are discussed. Responses to transmembrane sequence may depend on protein-translocon interactions, but responses to charge difference may be mediated by the electrostatic field provided by anionic phospholipids.

Specimens were formaldehyde fixed, paraffin embedded and hematoxylin and eosin (H&E) stained. Representative sections were reviewed by a pathologist (CS) for histologic subtype, grade, staging and percent tumor burden. Sections were macrodissected to increase the representation of tumor DNA in the total DNA extracted from the specimen. DNA was isolated following cell lysis and proteinase K treatment using the QiaQuick extraction method (Qiagen, VHilden Germany). Analysis of microsatellite instability (MSI)DNA from normal and tumor specimens was amplified using the MSI analysis system version 1.2 (Promega, Madison WI, USA) according to the manufacturer's instructions. Amplicons were detected using capillary electrophoresis on an ABI 3130xl Genetic Analyzer (Life Technologies, Carlsbad CA, USA) and the results were analyzed using GeneMapper V3.7 software (Life Technologies, Carlsbad CA, USA). The presence of instability in two, or more, of the five loci (>30%) was considered MSI-high (MSI-H). SNaPshot mutation profilingA SNaPshot single base extension assay was used to assess the mutation status of 62 loci in 7 genes (AKT1, BRAF, KRAS, NRAS, PIK3CA, PTEN and SMAD4) associated with CRC prognosis and treatment (9). SNaPshot products were separated using an ABI 3130xl Genetic Analyzer (Life Technologies, Carlsbad CA, USA) and compared to positive and normal controls for interpretation. Massively parallel DNA sequencingMultiplex amplicon-based sequencing libraries were prepared using the GeneRead DNA-seq Human Comprehensive Cancer Panel NGHS-501X (Qiagen, Hilden Germany) following the manufacturer's instructions. This panel targets coding and UTR regions of 124 commonly mutated genes in multiple cancer types. Once prepared, libraries were sequenced using a MiSeq 300 cycle V2 reagent kit (Illumina, San Diego CA, USA) with MiSeq Control software V2.3.0.3 and RTA software V1.18.42.0. Variant analysis was performed using the CLCbio genomics workbench (Qiagen, Hilden Germany). ResultsAn 81-year-old Egyptian male presented with weight loss, upper quadrant abdominal and rectal pain and blood streaked stools. The medical history was significant for cancer including a tumor of unreported origin removed by a partial small bowel resection in his 50's, renal cell carcinoma removed by nephrectomy in his 60's and prostate cancer treated with implantation of radioactive seeds in his 70's. The family history was also significant for cancer. Two first-degree relatives had colon cancer, 2 first-degree relatives had kidney cancer and a first-degree relative had an unspecified lymph node/head and neck cancer.After total proctocolectomy, 6 lesions were identified. These included 5 colonic and 1 rectal lesion. Lesion 1 (medullary carcinoma, 5 cm in greatest dimension, T4N0M0) was located in the left colon near to the splenic flexure ( Figure 1A). Lesion 2 (moderately differentiated invasive adenocarcinoma, 6 cm, T3N0M0) was located in the right colon near to the hepatic flexure ( Figure 1B). The colon also contained 3 early invasive carcin...

One approach to water optimization in agriculture is to increase soil water retention. A soil amendment that has received attention for its ability to increase soil water retention is biochar, the remaining biomass after high-C materials have been pyrolyzed. Two studies were conducted in Utah at two sites each during 2018-2020 to evaluate how a fast-pyrolysis wood biochar influenced the productivity and crop quality of irrigated alfalfa (Medicago sativa L.), corn (Zea mays L.), and wheat (Triticum aestivum L.) along with soil water tension. One study included a single biochar rate of 22 Mg ha −1 at two levels of irrigation (full vs. partial), where soil-incorporated biochar increased silage corn yield by 12% in 2018 and reduced yield by 10% in 2019. Topdressed biochar had no effect on alfalfa yield from 2018 to 2020 in either irrigation rate. The other trial had six to seven biochar rates (0-67 Mg ha −1 ) plus the addition of wood chips at a single rate (22 Mg ha −1 ) at one site. This trial displayed a yield decrease of 0.01 Mg ha −1 of wheat grain for each additional 1 Mg ha −1 of biochar applied; however, there were no effects on silage corn production. Data obtained over 10 site-years showed minimal effects from this type of biochar on crop yield, quality, or soil water tension, leading us to conclude that this wood biochar was not an effective tool for enhancing short-term crop production or conserving irrigation water in arid agriculture.

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