Thermal stress affects proliferation and differentiation of turkey satellite cells through the mTOR/S6K pathway in a growth-dependent manner

Satellite cells (SCs) are stem cells responsible for post-hatch muscle growth through hypertrophy and in birds are sensitive to thermal stress during the first week after hatch. The mechanistic target of rapamycin (mTOR) signaling pathway, which is highly responsive to thermal stress in differentiating turkey pectoralis major (p. major) muscle SCs, regulates protein synthesis and the activities of SCs through a downstream effector, S6 kinase (S6K). The objectives of this study were: 1) to determine the effect of heat (43°C) and cold (33°C) stress on activity of the mTOR/S6K pathway in SCs isolated from the p. major muscle of one-week-old faster-growing modern commercial (NC) turkeys compared to those from slower-growing Randombred Control Line 2 (RBC2) turkeys, and 2) to assess the effect of mTOR knockdown on the proliferation, differentiation, and expression of myogenic regulatory factors of the SCs. Heat stress increased phosphorylation of both mTOR and S6K in both turkey lines, with greater increases observed in the RBC2 line. With cold stress, greater reductions in mTOR and S6K phosphorylation were observed in the NC line. Early knockdown of mTOR decreased proliferation, differentiation, and expression of myoblast determination protein 1 and myogenin in both lines independent of temperature, with the RBC2 line showing greater reductions in proliferation and differentiation than the NC line at 38° and 43°C. Proliferating SCs are more dependent on mTOR/S6K-mediated regulation than differentiating SCs. Thus, thermal stress can affect breast muscle hypertrophic potential by changing satellite cell proliferation and differentiation, in part, through the mTOR/S6K pathway in a growth-dependent manner. These changes may result in irreversible effects on the development and growth of the turkey p. major muscle.

Genome-Wide Investigation of the Cysteine Synthase Gene Family Shows That Overexpression of CSase Confers Alkali Tolerance to Alfalfa (Medicago sativa L.)

Alfalfa is widely grown worldwide as a perennial high-quality legume forage and as a good ecological landcover. The cysteine synthase (CSase) gene family is actively involved in plant growth and development and abiotic stress resistance but has not been systematically investigated in alfalfa. We identified 39 MsCSase genes on 4 chromosomes of the alfalfa genome. Phylogenetic analysis demonstrated that these genes were clustered into six subfamilies, and members of the same subfamily had similar physicochemical properties and sequence structures. Overexpression of the CSase gene in alfalfa increased alkali tolerance. Compared with control plants, the overexpression lines presented higher proline, soluble sugars, and cysteine and reduced glutathione contents and superoxide dismutase and peroxidase activities as well as lower hydrogen peroxide and superoxide anion contents after alkali stress. The relative expression of γ-glutamyl cysteine synthetase gene (a downstream gene of CSase) in the overexpression lines was much higher than that in the control line. The CSase gene enhanced alkalinity tolerance by regulating osmoregulatory substances and improving antioxidant capacity. These results provide a reference for studying the CSase gene family in alfalfa and expanding the alkali tolerance gene resources of forage plants.

Performance Evaluation of the BZ COVID-19 Neutralizing Antibody Test for the Culture-Free and Rapid Detection of SARS-CoV-2 Neutralizing Antibodies

Rapid and accurate measurement of SARS-CoV-2 neutralizing antibodies (nAbs) can aid in understanding the development of immunity against COVID-19. This study evaluated the diagnostic performance of a rapid SARS-CoV-2 nAb detection test called the BZ COVID-19 nAb test BZ-nAb (BZ-nAb; BioZentech). Using the 90% plaque-reduction neutralization test (PRNT-90) as a reference, 104 serum specimens collected from COVID-19-positive and -negative patients were grouped into 40 PRNT-90-positive and 64 PRNT-90-negative specimens. The performance of the BZ-nAb was compared with that of the cPass surrogate virus neutralization test (cPass sVNT; Genscript). The BZ-nAb showed a sensitivity ranging from 92.5%–95.0% and specificity ranging from 96.9%–100%, whereas cPass sVNT showed a sensitivity of 100% (95% confidence interval (CI) 90.5%–100%) and specificity of 98.4% (95% CI, 91.6%–100%). The dilution factor obtained with PRNT-90 showed a stronger correlation with the percent inhibition of cPass sVNT (r = 0.8660, p < 0.001) compared with the test and control line ratio (T/C ratio) of the BZ-nAb (r = −0.7089, p < 0.001). An almost perfect agreement was seen between the BZ-nAb and cPass sVNT results, with a strong negative correlation between the BZ-nAb T/C ratio and cPass sVNT percent inhibition (r = −0.8022, p < 0.001). In conclusion, the diagnostic performance of the BZ-nAb was comparable to that of the cPass sVNT, although the BZ-nAb had a slightly lower sensitivity.

Obtaining Salt Stress-Tolerant Eggplant Somaclonal Variants from In Vitro Selection

An efficient regeneration protocol was applied to regenerate shoots on salt stress-tolerant calli lines of aubergine (Solanum melongena). These NaCl-tolerant cell lines were obtained by two different methods. On the one hand, the developed callus tissue was transferred to a medium with a continuous salt content of 40, 80, 120, or 160 mM NaCl. On the other hand, the callus tissue was subjected to a stepwise increasing salinity to 160 mM NaCl every 30 days. With the second method, calli which could be selected were characterized by compact growth, a greenish color, and absence of necrotic zones. When grown on salt-free medium again, NaCl-tolerant calli showed a decline in relative growth rate and water content in comparison to the control line. This was more obvious in the 120 mM NaCl-tolerant callus. Lipid peroxidase activity increased in 40 and 80 mM NaCl-tolerant calli; yet did not increase further in 120 mM-tolerant callus. An increase in ascorbic acid content was observed in 80 and 120 mM NaCl-tolerant calli compared to the 40 mM NaCl-tolerant lines, in which ascorbic acid content was twice that of the control. All NaCl-tolerant lines showed significantly higher superoxide dismutase (SOD) (208–305–370 µmol min−1 mg−1 FW) and catalase (CAT) (136–211–238 µmol min−1 mg−1 FW) activities compared to control plants (231 and 126 µmol min−1 mg−1 FW). Plants were regenerated on the calli lines that could tolerate up to 120 mM NaCl. From the 32 plants tested in vitro, ten plants with a higher number of leaves and root length could be selected for further evaluation in the field. Their high salt tolerance was evident by their more elevated fresh and dry weight, their more increased relative water content, and a higher number and weight of fruits compared to the wild-type parental control. The presented work shows that somaclonal variation can be efficiently used to develop salt-tolerant mutants.

A Selected Population Study Reveals the Biochemical Mechanism of Intramuscular Fat Deposition in Chicken Meat

Background Increasing intramuscular fat (IMF) is an important strategy to improve meat quality, but the regulation mechanism of IMF deposition needs to be systematically clarified. Results A total of 520 chickens from a selected line with improved IMF content and a control line were used to investigate the biochemical mechanism of IMF deposition in chickens. The results showed that the increased IMF would improve the flavor and tenderness quality of chicken meat. IMF content was mainly determined both by measuring triglyceride (TG) and phospholipid (PLIP) in muscle tissue, but only TG content was found to be decisive for IMF deposition. Furthermore, the increase in major fatty acid (FA) components in IMF is mainly derived from TGs (including C16:0, C16:1, C18:1n9c, and C18:2n6c, etc.), and the inhibition of certain very-long-chain FAs would help to IMF/TG deposition. Conclusions Our study elucidated the underlying biochemical mechanism of IMF deposition in chicken: Prevalent accumulation of long-chain FAs and inhibitions of medium-chain FAs and very long chain-FA would synergistically result in the increase of TGs with the FA biosynthesis and cellular uptake ways. Our findings will guide the production of high-quality chicken meat.

Smart Segmented Polymer Injection Pilot: A Case Study

Polymer flood is proved an effective method for EOR in China. Traditional segmented polymer injection technique cannot obtain continuous layer parameters. Real-time monitoring is necessary for polymer flood because downhole pressure and flowrate vary more often than waterflood. Existing technique for layered monitoring and flowrate adjustment is wireline test. There is no smart technique which can realize real-time monitoring and automatic flowrate control. In this paper, a smart segmented injection technique for polymer flood well is introduced. A smart distributor is permanently placed in each layer. It is composed of flowmeter, temperature sensor, two pressure sensors, downhole choke and electrical control unit. The special flowmeter is adopted for polymer flowrate test. All the distributors are connected together by a single control line which is set outside of the tubing string. Operator can read the data of each layer and adjust the flowrate whenever needed at any time which makes the technique a smart one. The smart technique for polymer flood wells has been implemented in a polymer well in Daqing oilfield of China. A case study for smart segmented polymer injection pilot is introduced in detail including technical principle, indoor test results, construction process and adjustment process. The application results show that the operator on the ground can easily obtain downhole tubing pressure, layer annulus pressure, temperature and flowrate on line. The sample time can be set to any one between 1-65536s according to geological engineer's advice. There is no limitation caused by battery power because the distributor is powered by cable on the ground. In terms of adjustment, the flowrate can be adjusted according to the target value. And it can also be regulated at any time manually, just needing pushing the mouse in the office. The application also displays that the smart segmented technique has the advantage for polymer injection because of larger change of layered parameters. It can provide more real-time data for oil development engineer and the data are beneficial for better understanding and optimization of the reservoir. Therefore, the smart segmented polymer injection has a great potential for EOR based on polymer flood.

Flawless Cutting of Tubing and Control Lines by Mechanical Pipe Cutter in Challenging Well Condition Provides an Environmental Friendly Alternative: A Case Study from Brunei

The Electro-Mechanical Pipe Cutter (MPC) is a non-ballistic & non-chemical wireline deployed alternative cutter tool for parting downhole tubular in the process of well abandonment, pipe recovery and retrieving of packer elements. This case study showcases its application in two wells with different challenges in cutting 4.5" tubing viz., (i) with multiple control lines to facilitate fishing operations and (ii) under compression in a highly deviated trajectory. In Well A, Brunei offshore, the position of the tubing and orientation of the control lines were challenging for ballistic option, along with the possibility of scarring the 9-7/8" casing during the cutting operation. Thus, 3-1/8" OD MPC was used for this job to cut near the coupling, ensuring optimum stand off from casing wall aiming to achieve cutting the control lines in tension. Dual cut were designed to allow the room for a safe cut zone. The primary cut was performed near middle of the joint at ∼1985m, with the tubing in tension. The cut was initiated at a very slow feed (0.2 mm/min) and motor rates (4000rpm), which was gradually increased once the cutting was stable. After the accomplishment of the tubing cut, the parameters were again reduced to carefully cutting through control line. The tubing was successfully retrieved with smooth cut without any over pull indicating it to be completely free. The flawless cutting operation was performed in less than one hour with outmost efficiency. In another highly inclined Well B, Brunei offshore, MPC was chosen over ballistic because it was needed to be conveyed by tractor and ballistic shock has potential to damage it during the operation. Also the advantage of MPC to perform multiple cuts in one run, made it a preferred choice. In this well, multiple cuts were performed to weaken the joint connection of the tubing to allow the rig to pull it free. It was to overcome the adversity posed by high inclination and the pipe under compression. Three cuts were performed at ∼2996 m, each 20 cm apart with an OD of nearly 4.609". After completion of the job, the circulation was performed with surface return, indicating successful execution and the tubing was retrieved on surface showing a clean cut. This case study shows the appropriate planning and execution of the mechanical pipe cutter can provide an efficient, environment friendly and safe alternative to cut tubing and control line in the challenging condition especially when an explosive and chemical cutter options are not considered suitable.

Journey of Effectively Using Real-Time Production Surveillance Tool in Digital Transformation and Well, Reservoir and Facility Management Improvements

In 2014, SPE-167857-MS was published highlighting how Real-Time Surveillance using the Shell proprietary Production Universe (PU) application helped to reduce deferment, improve production allocation, optimize test unit capacity, and track well operating envelope in Brunei Shell Petroleum BSP (company) operations in South East Asia. Since then, there has been significant progress in the application of PU to help the company meet Wells Reservoir Management (WRFM) requirements and Operational Excellence standards in areas such as Platform Production Reconciliation, Well Modeling, Production Estimation, and Exception based Surveillance helping the company to improve their Hydrocarbon and Energy Accounting. The wider introduction of the PU application in the allocation process significantly helped in Hydrocarbon Accounting (HCA), helping company's journey in moving from monthly to daily allocation and assisting to improve the field reconciliation factor (RF). The utilization of PU has also facilitated real-time monitoring of production parameters supporting engineers to safely and efficiently operate their wells within the Operating Envelopes while adhering to reservoir management guidelines. The optimization engine of the PU has been used to maximize the production of company contributing to two major success stories of Real-Time Condensate Optimization in a Gas-Constraint system and Gas Lift Optimization in a platform with limited lift gas availability amongst the producing wells. An Integrated Production Monitoring and Optimization System (IPMOS) provides asset-wide advice on optimally producing company's well within the constraints imposed by limitations on pipeline capacity, compressor throughput, and remote operability while satisfying customer demands. PU is additionally being used in Proactive Technical Monitoring (PTM) of rotating equipment to identify the critical parameters operating outside the set limits in an exception-based format. PU alerts & alarms have been configured in a wide range of operational monitoring such as Ensure Safe Production (ESP), Chemical Injection, Annulus Pressure Monitoring(APM), Control-Line pressure, Erosion-Corrosion Monitoring System(ECMS) to alert Production Engineers in case of any discrepancy or exception-based format for them to take remedial actions. This paper will explain how each of the above applications in PU has helped company in its journey of closing its gap to potential and achieving the digital transformation of its operations.