Notes

The facial skin Picture Meta-Database (fIMDb) & ChatLab Face Anomaly Data source (CFAD): Equipment with regard to investigation in confront perception as well as social judgment.
Ribosome profiling, first developed in 2009, is the gold standard for quantifying and qualifying changes to translation genome-wide (Ingolia et al., Science, 2009). Though first designed and optimized in vegetative budding yeast, it has since been modified and specialized for use in diverse cellular states in yeast, as well as in bacteria, plants, human cells, and many other organisms (Ingolia et al. Science, 2009, reviewed in (Ingolia et al., Cold Spring Harb Perspect Biol, 2019; Brar and Weissman, Nat Rev Mol Cell Biol, 2015)). Here we report the current ribosome profiling protocol used in our lab to study genome-wide changes to translation in budding yeast undergoing the developmental process of meiosis (Brar et al., Science, 2012; Cheng et al., Cell, 2018). We describe this protocol in detail, including the following steps collection and flash freezing samples, cell lysis and extract preparation, sucrose gradient centrifugation and monosome collection, RNA extraction, library preparation, and library quality control. Almost every step presented here should be directly applicable to performing ribosome profiling in other eukaryotic cell types or cell states.Monitoring whole-genome translation and mRNA ribosome occupancy in vivo using ribosome profiling has proven to be a powerful tool for discovery of gene expression regulation, mechanisms of translation, and new open reading frames, in a wide range of different cell types in different organisms. Here we describe its application to the malaria parasite, Plasmodium falciparum. We present methods for intact polysome purification from parasite cultures, polysome digestion, monosome purification, ribosome footprint nucleic acid extraction, and Illumina library preparation.The knowledge of translation start sites is crucial for annotation of genes in bacterial genomes. However, systematic mapping of start codons in bacterial genes has mainly relied on predictions based on protein conservation and mRNA sequence features which, although useful, are not always accurate. We recently found that the pleuromutilin antibiotic retapamulin (RET) is a specific inhibitor of translation initiation that traps ribosomes specifically at start codons, and we used it in combination with ribosome profiling to map start codons in the Escherichia coli genome. This genome-wide strategy, that was named Ribo-RET, not only verifies the position of start codons in already annotated genes but also enables identification of previously unannotated open reading frames and reveals the presence of internal start sites within genes. Here, we provide a detailed Ribo-RET protocol for E. coli. Ribo-RET can be adapted for mapping the start codons of the protein-coding sequences in a variety of bacterial species.Modern DNA sequencing technologies have allowed for the sequencing of tens of thousands of bacterial genomes. While this explosion of information has brought about new insights into the diversity of the prokaryotic world, much less is known of the identity of proteins encoded within these genomes, as well as their rates of production. The advent of ribosome profiling, or the deep sequencing of ribosome-protected footprints, has recently enabled the systematic evaluation of every protein-coding region in a given experimental condition, the rates of protein production for each gene, and the variability in translation rates across each message. Here, I provide an update to the bacterial ribosome profiling approach, with a particular emphasis on a simplified strategy to reduce cloning time.The production of peptides as active pharmaceutical ingredients (APIs) by recombinant technologies is of emerging interest. A reliable production platform, however, is still missing due the inherent characteristics of peptides such as proteolytic sensitivity, aggregation and cytotoxicity. We have developed a new technology named Numaswitch solving present limitations. Numaswitch was successfully employed for the production of diverse peptides and small proteins varying in length, physicochemical and functional characteristics, including Teriparatide, Linaclotide, human β-amyloid and Serum amyloid A3. Additionally, the potential of Numaswitch for a cost-efficient commercial production is demonstrated yielding > 2 g Teriparatide per liter fermentation broth in a quality meeting API standard.Co-utilization of xylose and glucose and subsequent fermentation using Saccharomyces cerevisiae could enhance ethanol productivity. Directed engineering approaches have met with limited success due to interconnectivity of xylose metabolism with other intrinsic, hidden pathways. Therefore, random approaches like protoplast fusion were used to reprogram unidentified mechanisms. Saccharomyces cerevisiae LN, the best hexose fermenter, was fused with xylose fermenting Pichia stipitis NCIM 3498. Adavivint Protoplasts prepared using glucanex were fused under electric impulse and fusants were selected using 10% ethanol and cycloheximide (50 ppm) markers. Two fusants, 1a.23 and 1a.30 showing fast growth on xylose and tolerance to 10% ethanol, were selected. Higher extracellular protein expression observed in fusants as compared to parents was corroborated by higher number of bands resolved by two-dimensional analysis. Overexpression of XYL1, XYL2, XKS, and XUT4 in fusants as compared to S. cerevisiae LN as observed by RT-PCR analysis was substantiated by higher specific activities of XR, XDH, and XKS enzymes in fusants. During lignocellulosic hydrolysate fermentation, fusants could utilize glucose faster than the parent P. stipitis NCIM 3498 and xylose consumption in fusants was higher than S. cerevisiae LN.The biorefinery technology aiming at protein extraction is rising and identification of suitable plant biomass input with valuable protein compounds for extraction is needed. Forage crops have been evaluated by the Cornell Net Carbohydrate and Protein System (CNCPS), and the result used as proxy of extractable protein in a biorefinery process. This serves as a helpful link between crop production and refinery output; however, the method has never been validated. Such validation is the main aim of this study. Five forage species-white clover, red clover, lucerne, perennial ryegrass, and tall fescue-were cut at four dates during spring and processed in a lab-scale refinery (screw press and subsequent protein precipitation from the green juice). The pulp fraction and the precipitated protein concentrate were both CNCPS analyzed to follow the initial crude protein (CP) plant input into these two fractions. Total recovery in concentrate was highest for the legumes, which points to an advantage of these species in protein extraction setups. High recovery of B1 and B2 (50% or higher for the grasses) in the pulp demonstrated a large proportion of soluble protein ending up in the fibrous pulp and shed light on the reason behind high feed quality of the pulp fraction. In conclusion, the existing tentative assumption of extractable protein being equal to CNCPS fractions of B1 and B2 and partly B3 was shown to be too simplified. The presented findings can improve crop species screening in terms of expected extractable protein yield.
Ascertaining the origin of large tumors located in the region of the pancreas head and adjacent mesocolon can pose a challenge preoperatively. En bloc pancreatoduodenectomy with hemicolectomy is often required towards curative tumor resection (R0) of malignant tumors in this region.

Herein we report a case of a 48-year-old man with two contiguous masses each 5cm in size, located in the pancreatic head. The masses were detected incidentally by abdominal ultrasonography at an annual health check. Endoscopic biopsies revealed inflammation with no malignancy. Cross-sectional imaging showed the tumor direct invasion of the uncinate process of the pancreas, and the third portion of the duodenum. Based on imaging, a malignant submucosal tumor originating from mesenchymal cells in the mesentery of the transverse colon was made preoperatively. The mass required en bloc pancreatoduodenectomy, right hemicolectomy, and resection of the superior mesenteric vein. The final pathology was carcinosarcoma of the transverse colon. The patient survived 18years after surgery without recurrence.

Malignant tumors located in the region of the pancreas head should be considered for an en bloc curative tumor resection and adjuvant chemotherapy treatments offered that might be beneficial for carcinosarcoma.
Malignant tumors located in the region of the pancreas head should be considered for an en bloc curative tumor resection and adjuvant chemotherapy treatments offered that might be beneficial for carcinosarcoma.Two polypropylene HVAC electret filters a regular filter and an antimicrobial filter containing zinc pyrithione (ZPT), were compared for filtration performance. The study was conducted over 7 months in realistic conditions with semi-urban outdoor air. Several parameters were monitored over the study period the average temperature was about 20 °C and relative humidity about 60%, the average inlet concentration of cultivable microorganisms was 50 CFU m-3, the average inlet concentration of particles was 10 μg m-3, the filter pressure drop increased moderately by about 30 Pa, and the particle collection efficiency of soda fluorescein (median diameter 0.35 μm) decreased in the first half of the study period by about 30% and then stabilized. The microbial concentration on the filters was quantified every 2 months using an innovative methodology based on media coupons in conjunction with microorganism quantification by CFU counting, with 5 culture media favorable to bacteria and/or fungi growth. The microbial concentrations on the filters were between 100 and 2000 CFU cm-2. The antimicrobial effect of zinc pyrithione was confirmed by the fungi cultivated with DRBC agar no effects in the level of filter clogging were revealed in the range studied. The high statistical deviation in the results regarding the inhibiting effect of zinc pyrithione on bacteria prevents any conclusion.Cadmium (Cd) is a transition metal that is toxic to living organisms in the environment and endangers living organisms. To explore whether Cd induces apoptosis in pig thymus and its possible mechanism, the role Cd induction of the PTEN/PI3K/Akt pathway in apoptosis of thymus cells was studied in pigs. We found that Cd exposure (the feed is treated with Cd) significantly increased Cd accumulation in the thymus of pigs. The TUNEL assay confirmed the typical apoptotic characteristics of thymus in Cd group. Moreover, in the Cd group, the activities of antioxidant indices decreased significantly, while the levels of oxidative stress indexes increased significantly, and the mRNA levels of GSH, CAT, Gpx1, GST, SOD1, and SOD2 decreased obviously. link2 Moreover, the mRNA and protein levels of PTEN/PI3K/AKT and apoptosis-related genes were detected by qPCR and western blotting. The results show that the expressions of PI3K and AKT decreased, while the expression of PTEN increased, indicating that pathway activated. With the PTEN/PI3K/AKT pathway regulating, Bcl-2 expression decreased. Conversely, the mRNA and protein expression of apoptosis-related genes were up-regulated. link3 In conclusion, accumulation of Cd in the pigs caused oxidative damage to immune tissues. In addition, Cd-induced oxidative stress activates the PTEN/PI3K/AKT pathway, inducing apoptosis in the thymus of pigs.
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