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Heme oxygenase-1-mediated apoptosis under cadmium-induced oxidative stress is regulated by autophagy, which is sensitized by tumor suppressor p53

Heme oxygenase-1 (HO-1) is a stress-inducible cytoprotective enzyme. It is often overexpressed in different types of cancers and promotes cell survival. However, the role of HO-1 and the underlying molecular mechanism of cadmium (Cd)-induced oxidative stress in cancer cells remain undefined. Here we show that the role of HO-1 under Cd-induced oxidative stress is dependent upon autophagy, which is sensitized by the tumor suppressor p53. The sensitivity to Cd was 3.5- and 14-fold higher in p53-expressing YD8 and H460 cells than in p53-null YD10B and H1299 cells, respectively. The levels of p53 in YD8 and H460 cells decreased in a Cd concentration-dependent manner, which was inhibited by pretreatment with N-acetylcysteine. In both cell lines, Cd exposure resulted in caspase-3-mediated PARP-1 cleavage and the induction of CHOP, LC3-II, and HO-1, which were limited in YD10B and H1299 cells exposed to high concentrations of Cd. Cd exposure to p53-overexpressing YD10B cells enhanced Cd-induced HO-1 and LC3-II levels, whereas genetic knockdown of p53 in YD8 cells resulted in the suppression of Cd-induced levels of HO-1 and LC3-II, indicating that p53 is required in the sensing of HO-1 and induction of autophagy. The inhibition of autophagy using small interfering RNA (siRNA) for the autophagy-related gene atg5 enhanced HO-1, CHOP, and PARP-1 cleavage induced by Cd. However, transfection with HO-1 siRNA increased Cd-induced LC3-II, and suppressed the expression of CHOP and cleavage of PARP-1. Collectively, the role of HO-1 in apoptosis could be modulated by autophagy, which is sensitized by p53 expression in human cancer cell lines. - Highlights: • Cadmium exposure decreased p53 level, and induced HO-1, apoptosis, and autophagy. • p53 sensitized Cadmium-induced HO-1 and autophagy induction. • Cadmium induced HO-1 under autophagy impairment and increased apoptosis. • Cadmium-induced autophagy was enhanced under HO-1 impaired conditions. • The role of HO-1 in Cadmium-induced apoptosis is modulated by autophagy.

60 APPLIED LIFE SCIENCES↗

Anthracycline resistance mediated by reductive metabolism in cancer cells: The role of aldo-keto reductase 1C3

Pharmacokinetic drug resistance is a serious obstacle that emerges during cancer chemotherapy. In this study, we investigated the possible role of aldo-keto reductase 1C3 (AKR1C3) in the resistance of cancer cells to anthracyclines. First, the reducing activity of AKR1C3 toward anthracyclines was tested using incubations with a purified recombinant enzyme. Furthermore, the intracellular reduction of daunorubicin and idarubicin was examined by employing the transfection of A549, HeLa, MCF7 and HCT 116 cancer cells with an AKR1C3 encoding vector. To investigate the participation of AKR1C3 in anthracycline resistance, we conducted MTT cytotoxicity assays with these cells, and observed that AKR1C3 significantly contributes to the resistance of cancer cells to daunorubicin and idarubicin, whereas this resistance was reversible by the simultaneous administration of 2′-hydroxyflavanone, a specific AKR1C3 inhibitor. In the final part of our work, we tracked the changes in AKR1C3 expression after anthracycline exposure. Interestingly, a reciprocal correlation between the extent of induction and endogenous levels of AKR1C3 was recorded in particular cell lines. Therefore, we suggest that the induction of AKR1C3 following exposure to daunorubicin and idarubicin, which seems to be dependent on endogenous AKR1C3 expression, eventually might potentiate an intrinsic resistance given by the normal expression of AKR1C3. In conclusion, our data suggest a substantial impact of AKR1C3 on the metabolism of daunorubicin and idarubicin, which affects their pharmacokinetic and pharmacodynamic behavior. In addition, we demonstrate that the reduction of daunorubicin and idarubicin, which is catalyzed by AKR1C3, contributes to the resistance of cancer cells to anthracycline treatment. - Highlights: • Metabolism of anthracyclines by AKR1C3 was studied at enzyme and cellular levels. • Anthracycline resistance mediated by AKR1C3 was demonstrated in cancer cells. • Induction of AKR1C3 after anthracycline exposure was investigated in cancer cells. • AKR1C3 confers resistance of cancer cells to daunorubicin and idarubicin. • AKR1C3 can be induced by the exposure to anthracyclines in some cell lines.

60 APPLIED LIFE SCIENCES↗

Isothermal Ice Crystallization Kinetics in the Gas-Diffusion Layer of a Proton-Exchange-Membrane Fuel Cell

Nucleation and growth of ice in the fibrous gas-diffusion layer (GDL) of a proton-exchange membrane fuel cell (PEMFC) are studied using isothermal differential scanning calorimetry (DSC). Isothermal crystallization rates and pseudo-steady-state nucleation rates are obtained as a function of subcooling from heat-flow and induction-time measurements. Kinetics of ice nucleation and growth are studied at two polytetrafluoroethylene (PTFE) loadings (0 and 10 wt %) in a commercial GDL for temperatures between 240 and 273 K. A nonlinear ice-crystallization rate expression is developed using Johnson–Mehl–Avrami–Kolmogorov (JMAK) theory, in which the heat-transfer-limited growth rate is determined from the moving-boundary Stefan problem. Induction times follow a Poisson distribution and increase upon addition of PTFE, indicating that nucleation occurs more slowly on a hydrophobic fiber than on a hydrophilic fiber. The determined nucleation rates and induction times follow expected trends from classical nucleation theory. Finally, a validated rate expression is now available for predicting ice-crystallization kinetics in GDLs.

30 DIRECT ENERGY CONVERSION↗

High quality factor, fully switchable terahertz superconducting metasurface

We present a complementary THz metasurface realised with Niobium thin film which displays a quality factor Q = 54 and a fully switchable behaviour as a function of the temperature. The switching behaviour and the high quality factor are due to a careful design of the metasurface aimed at maximising the ohmic losses when the Nb is above the critical temperature and minimising the radiative coupling. The superconductor allows the operation of the cavity with high Q and the use of inductive elements with a high aspect ratio. Comparison with three dimensional finite element simulations highlights the crucial role of the inductive elements and of the kinetic inductance of the Cooper pairs in achieving the high quality factor and the high field enhancement.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Dynamic hysteretic sensing model of bending-mode Galfenol transducer

A dynamic hysteretic sensing model has been developed to predict the dynamic responses of the magnetic induction, the stress, and the output voltage for a bending-mode Galfenol unimorph transducer subjected simultaneously to acceleration and bias magnetic field. This model is obtained by coupling the hysteretic Armstrong model and the structural dynamic model of the Galfenol unimorph beam. The structural dynamic model of the beam is founded based on the Euler-Bernouli beam theory, the nonlinear constitutive equations, and the Faraday law of electromagnetic induction. Comparisons between the calculated and measured results show the model can describe dynamic nonlinear voltage characteristics of the device, and can predict hysteretic behaviors between the magnetic induction and the stress. Moreover, the model can effectively analyze the effects of the bias magnetic field, the acceleration amplitude, and frequency on the root mean square voltage of the device.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Microwave dynamics of high aspect ratio superconducting nanowires studied using self-resonance

We study the microwave impedance of extremely high aspect ratio (length/width ≈ 5000) superconducting niobium nitride nanowires. The nanowires are fabricated in a compact meander geometry that is in series with the center conductor of a 50 Ω coplanar waveguide transmission line. The transmission coefficient of the sample is measured up to 20 GHz. At high frequency, a peak in the transmission coefficient is seen. Numerical simulations show that this is a half-wave resonance along the length of the nanowire, where the nanowire acts as a high impedance, slow wave transmission line. This resonance sets the upper frequency limit for these nanowires as inductive elements. Fitting simulations to the measured resonance enables a precise determination of the nanowire's complex sheet impedance at the resonance frequency. The real part is a measure of dissipation, while the imaginary part is dominated by kinetic inductance. We characterize the dependence of the sheet resistance and sheet inductance on both temperature and current and compare the results to recent theoretical predictions for disordered superconductors. These results can aid in the understanding of high frequency devices based on superconducting nanowires. They may also lead to the development of novel superconducting devices such as ultra-compact resonators and slow-wave structures.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

3-D Prismatic Packaging Methodologies for Wide Band Gap Power Electronics Modules

In a power module the parasitic inductance limits the dynamic high-frequency performance, and the area of the cooling surfaces limits the power capability. This paper presents a new 3D power electronic design methodology based on the concept of mutual inductance cancellation and multi-sided heat transfer. New 3D prismatic packaging concepts are proposed for Wide Band Gap (WBG) power devices, where the devices can be mounted at acute angles to adjacent interconnects or other devices. Discussion is given on electrical and thermal path optimization in a 3D space. To validate the 3D prismatic packaging methodology, a 1200V/50A SiC half-bridge power module is fabricated and tested for electrical and thermal performance and results are compared with simulations. The power density calculated for the module is 12 kW/in3 (including heatsink) and shows a 31.3% inductance reduction compared to a 2D planar module. Finally, design guidance suggested for utilizing prismatic structures is provided, together with suggested future work in the area. This paper presents the first reported true 3D power module.

24 POWER TRANSMISSION AND DISTRIBUTION↗

AMAROK: A Radio Frequency Development Platform for High-Power, Full-Scale Positive Ion Sources for DIII-D Neutral Beam Injectors

Next-generation neutral beam injection (NBI) systems demand RF ion sources capable of efficiently coupling >120 kW at 2–4-MHz frequencies, yet existing designs face voltage standoff and impedance-matching challenges. To address this, the advanced multicoil antenna for RF operations at kilowatts (AMAROK) was developed as a high-power RF inductively coupled plasma (ICP) source delivering up to 200 kW in the 2–4-MHz range via four phase-controlled generators for flexible power sharing. Two antenna designs—a single-strap multiturn (MT) and a multistrap single-turn (ST) design—were evaluated to optimize resonance, impedance matching, and power coupling across plasma loads. Here, a semi-analytical self-resonant frequency (SRF) model, validated experimentally, predicts resonance trends for arbitrary turn counts and tubing diameters, enabling rapid antenna optimization. Strap-to-strap mutual inductance in the ST configuration showed strong spatial dependence, guiding generator operation and total load inductance. These insights informed a custom π -topology matching network, achieving stable impedance matching over a wide range of plasma-driven loads. Collectively, these results position AMAROK as a versatile testbed for advancing high-power RF source technology in fusion NBI applications.

DIII-D↗

Effect of Longitudinal Magnetic Field on the Spread of Erosion Laser Plasma in Vacuum and Generation of Plasma Magnetic Fields

Dynamics of erosion laser plasma in vacuum and generation of magnetic field by moving plasma (in particular, in the presence of external static magnetic field oriented along the direction of plasma motion) are experimentally studied. Radial confinement of the spread of plasma, a decrease in the electrification of target upon plasma formation, and an increase in the induction of the plasma magnetic field by a factor of 10–15 are revealed at an induction of the external magnetic field of about 0.35 T. Dependences of the induction of the plasma magnetic field on the power density of the laser radiation are determined for the above regimes.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Analysis Background & Noise in Stretched Wire Alignment Technique Measurements

The Stretched-Wire Alignment Technique (SWAT) is one method of magnet alignment for linear induction accelerators. The applications of SWAT have been implemented for aligning solenoid magnets on the Scorpius linear induction accelerator which will be sited at the Nevada National Security Site and the Flash X-Ray (FXR) linear induction accelerator at Lawrence Livermore National Laboratory’s Contained Firing Facility. This article describes both systematic (repeatable) and random sources of background and noise as well as practical ways to eliminate or reduce them to acceptable levels. Systematic sources include reflections from wire ends, rapid sag due to ohmic heating of the wire, magnetic materials, and shot rate. Random sources include air currents, vibration of nearby equipment, mechanical stability of test equipment, and the instruments used to measure the wire motion. Mitigations include curve fitting and adaptive noise signal cancellation, and mechanical damping. Finite Element Analysis (FEA) was used to identify and resolve a repeatable wire vibration frequency interfering with the signal resolution. Two stretched wire alignment technique set ups from Sandia National Labs and Lawrence Livermore National Lab have shown background noise sources and ways of mitigating them by either analysis methods or change of mechanical configuration. Conclusions that were drawn included the severe sensitivity of the deflection to even small external interferences of the SWAT wire such that it requires attention to detail in mechanical set up and analysis.

Linear Inductive Accelerator↗

LDRD final report on confinement of cluster fusion plasmas with magnetic fields.

Two versions of a current driver for single-turn, single-use 1-cm diameter magnetic field coils have been built and tested at the Sandia National Laboratories for use with cluster fusion experiments at the University of Texas in Austin. These coils are used to provide axial magnetic fields to slow radial loss of electrons from laser-produced deuterium plasmas. Typical peak field strength achievable for the two-capacitor system is 50 T, and 200 T for the ten-capacitor system. Current rise time for both systems is about 1.7 {mu}s, with peak current of 500 kA and 2 MA, respectively. Because the coil must be brought to the laser, the driver needs to be portable and drive currents in vacuum. The drivers are complete but laser-plasma experiments are still in progress. Therefore, in this report, we focus on system design, initial tests, and performance characteristics of the two-capacitor and ten-capacitors systems. The questions of whether a 200 T magnetic field can retard the breakup of a cluster-fusion plasma, and whether this field can enhance neutron production have not yet been answered. However, tools have been developed that will enable producing the magnetic fields needed to answer these questions. These are a two-capacitor, 400-kA system that was delivered to the University of Texas in 2010, and a 2-MA ten-capacitor system delivered this year. The first system allowed initial testing, and the second system will be able to produce the 200 T magnetic fields needed for cluster fusion experiments with a petawatt laser. The prototype 400-kA magnetic field driver system was designed and built to test the design concept for the system, and to verify that a portable driver system could be built that delivers current to a magnetic field coil in vacuum. This system was built copying a design from a fixed-facility, high-field machine at LANL, but made to be portable and to use a Z-machine-like vacuum insulator and vacuum transmission line. This system was sent to the University of Texas in Austin where magnetic fields up to 50 T have been produced in vacuum. Peak charge voltage and current for this system have been 100 kV and 490 kA. It was used this last year to verify injection of deuterium and surrogate clusters into these small, single-turn coils without shorting the coil. Initial test confirmed the need to insulate the inner surface of the coil, which requires that the clusters must be injected through small holes in an insulator. Tests with a low power laser confirmed that it is possible to inject clusters into the magnetic field coils through these holes without destroying the clusters. The university team also learned the necessity of maintaining good vacuum to avoid insulator, transmission line, and coil shorting. A 200-T, 2 MA system was also constructed using the experience from the first design to make the pulsed-power system more robust. This machine is a copy of the prototype design, but with ten 100-kV capacitors versus the two used in the prototype. It has additional inductance in the switch/capacitor unit to avoid breakdown seen in the prototype design. It also has slightly more inductance at the cable connection to the vacuum chamber. With this design we have been able to demonstrate 1 MA current into a 1 cm diameter coil with the vacuum chamber at air pressure. Circuit code simulations, including the additional inductance with the new design, agree well with the measured current at a charge voltage of 40 kV with a short circuit load, and at 50 kV with a coil. The code also predicts that with a charge voltage of 97 kV we will be able to get 2 MA into a 1 cm diameter coil, which will be sufficient for 200 T fields. Smaller diameter or multiple-turn coils will be able to achieve even higher fields, or be able to achieve 200-T fields with lower charge voltage. Work is now proceeding at the university under separate funding to verify operation at the 2-MA level, and to address issues of debris mitigation, measurement of the magnetic field, and operation in vacuum. We anticipate operation at full current with single-turn, magnetic field coils this fall, with 200 T experiments on the Texas Petawatt laser in the spring of 2012.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Continuing the Validation of CCIM Processability for Glass Ceramic HLLW Forms: Plan for Test AFY14CCIM-GC1

This test plan covers test AFY14CCIM-GC1which is the first of two scheduled FY-2014 test runs involving glass ceramic waste forms in the Idaho National Laboratory’s Cold Crucible Induction Melter Pilot Plant. The test plan is based on the successes and challenges of previous tests performed in FY-2012 and FY-2013. The purpose of this test is to continue to collect data for validating the glass ceramic High Level Liquid Waste form processability advantages using Cold Crucible Induction Melter technology. The major objective of AFYCCIM-GC1 is to complete additional proposed crucible pouring and post tapping controlled cooling experiments not completed during previous tests due to crucible drain failure. This is necessary to qualify that no heat treatments in standard waste disposal canisters are necessary for the operational scale production of glass ceramic waste forms. Other objectives include the production and post-test analysis of surrogate waste forms made from separate pours into the same graphite mold canister, testing the robustness of an upgraded crucible bottom drain and drain heater assembly, testing the effectiveness of inductive melt initiation using a resistive starter ring with a square wave configuration, and observing the tapped molten flow behavior in pans with areas identical to standard High Level Waste disposal canisters. Testing conditions, the surrogate waste composition, key testing steps, testing parameters, and sampling and analysis requirements are defined.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

In-situ Condition Monitoring of Components in Small Modular Reactors Using Process and Electrical Signature Analysis. Final report, volume 1. Development of experimental flow control loop, data analysis and plant monitoring

The research and development under this project was focused on the following three major objectives: Objective 1: Identification of critical in-vessel SMR components for remote monitoring and development of their low-order dynamic models, along with a simulation model of an integral pressurized water reactor (iPWR). Objective 2: Development of an experimental flow control loop with motor-driven valves and pumps, incorporating data acquisition and on-line monitoring interface. Objective 3: Development of stationary and transient signal processing methods for electrical signatures, machinery vibration, and for characterizing process variables for equipment monitoring. This objective includes the development of a data analysis toolbox. The following is a summary of the technical accomplishments under this project: - A detailed literature review of various SMR types and electrical signature analysis of motor-driven systems was completed. A bibliography of literature is provided at the end of this report. Assistance was provided by ORNL in identifying some key references. - A review of literature on pump-motor modeling and digital signal processing methods was performed. - An existing flow control loop was upgraded with new instrumentation, data acquisition hardware and software. The upgrading of the experimental loop included the installation of a new submersible pump driven by a three-phase induction motor. All the sensors were calibrated before full-scale experimental runs were performed. - MATLAB-Simulink model of a three-phase induction motor and pump system was completed. The model was used to simulate normal operation and fault conditions in the motor-pump system, and to identify changes in the electrical signatures. - A simulation model of an integral PWR (iPWR) was updated and the MATLAB-Simulink model was validated for known transients. The pump-motor model was interfaced with the iPWR model for testing the impact of primary flow perturbations (upsets) on plant parameters and the pump electrical signatures. Additionally, the reactor simulation is being used to generate normal operation data and data with instrumentation faults and process anomalies. A frequency controller was interfaced with the motor power supply in order to vary the electrical supply frequency. The experimental flow control loop was used to generate operational data under varying motor performance characteristics. Coolant leakage events were simulated by varying the bypass loop flow rate. The accuracy of motor power calculation was improved by incorporating the power factor, computed from motor current and voltage in each phase of the induction motor.- A variety of experimental runs were made for steady-state and transient pump operating conditions. Process, vibration, and electrical signatures were measured using a submersible pump with variable supply frequency. High correlation was seen between motor current and pump discharge pressure signal; similar high correlation was exhibited between pump motor power and flow rate. Wide-band analysis indicated high coherence (in the frequency domain) between motor current and vibration signals. - Wide-band operational data from a PWR were acquired from AMS Corporation and used to develop time-series models, and to estimate signal spectrum and sensor time constant. All the data were from different pressure transmitters in the system, including primary and secondary loops. These signals were pre-processed using the wavelet transform for filtering both low-frequency and high-frequency bands. This technique of signal pre-processing provides minimum distortion of the data, and results in a more optimal estimation of time constants of plant sensors using time-series modeling techniques.

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗

Comparison Between Numerical and Experimental Results on Mechanical Stirrer and Bubbling in a Cylindrical Tank - 13047

The process of vitrification in a cold crucible heated by direct induction is used in the fusion of oxides. Its feature is the production of high-purity materials. The high-level of purity of the molten is achieved because this melting technique excludes the contamination of the charge by the crucible. The aim of the present paper is to analyze the hydrodynamic of the vitrification process by direct induction, with the focus in the effects associated with the interaction between the mechanical stirrer and bubbling. Considering the complexity of the analyzed system and the goal of the present work, we simplified the system by not taking into account the thermal and electromagnetic phenomena. Based in the concept of hydraulic similitude, we performed an experimental study and a numerical modeling of the simplified model. The results of these two studies were compared and showed a good agreement. The results presented in this paper in conjunction with the previous work contribute to a better understanding of the hydrodynamics effects resulting from the interaction between the mechanical stirrer and air bubbling in the cold crucible heated by direct induction. Further works will take into account thermal and electromagnetic phenomena in the presence of mechanical stirrer and air bubbling. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Metabolomics of related C3 and C4 Flaveria species indicate differences in the operation of photorespiration under fluctuating light

C 3 photosynthesis can be complemented with a C 4 carbon concentrating mechanism (CCM) to minimize photorespiratory losses. C 4 photosynthesis is often more efficient than C 3 under steady-state conditions. However, the C 4 CCM depends on inter-cellular metabolite concentration gradients, which must increase following increases in light intensity and could decrease rates of C 4 photosynthesis under fluctuating light. Additionally, incomplete flux through photorespiration could prove beneficial to C 4 assimilation during light induction of the CCM. Here, we compare metabolic profiles in the closely related C 3 Flaveria robusta and C 4 Flaveria bidentis during a light transient from low to high light to determine if these non-steady state accumulation patterns provide insight to the induction of the metabolite gradients needed to drive C 4 intermediate transport and if there is incomplete cycling of photorespiratory intermediates. In these C 3 and C 4 species, metabolite steady-state pool sizes suggest that C 4 transport acids maintain concentration gradients across the bundle sheath and mesophyll cell types under these light fluctuations. However, there was incomplete flux through photorespiration in the C 4 F. bidentis, which could reduce photorespiratory CO 2 loss via glycine decarboxylation and help maintain higher rates of assimilation during following induction periods.

59 BASIC BIOLOGICAL SCIENCES↗

Unusual modifications of protein biomarkers expressed by plasmid, prophage, and bacterial host of pathogenic Escherichia coli identified using top‐down proteomic analysis

Rationale Pathogenic bacteria often carry prophage (bacterial viruses) and plasmids (small circular pieces of DNA) that may harbor toxin, antibacterial, and antibiotic resistance genes. Proteomic characterization of pathogenic bacteria should include the identification of host proteins and proteins produced by prophage and plasmid genomes. Methods Protein biomarkers of two strains of Shiga toxin–producingEscherichia coli(STEC) were identified using antibiotic induction, matrix‐assisted laser desorption/ionization tandem time‐of‐flight (MALDI‐TOF‐TOF) tandem mass spectrometry (MS/MS) with post‐source decay (PSD), top‐down proteomic (TDP) analysis, and plasmid sequencing. Alphafold2 was also used to compare predicted in silico structures of the identified proteins to prominent fragment ions generated using MS/MS‐PSD. Strain samples were also analyzed with and without chemical reduction treatment to detect the attachment of pendant groups bound by thioester or disulfide bonds. Results Shiga toxin was detected and/or identified in both STEC strains. For the first time, we also identified the osmotically inducible protein (OsmY) whose sequence unexpectedly had two forms: a full and a truncated sequence. The truncated OsmY terminates in the middle of an α‐helix as determined by Alphafold2. A plasmid‐encoded colicin immunity protein was also identified with and without attachment of an unidentified cysteine‐bound pendant group (~307 Da). Plasmid sequencing confirmed top‐down analysis and the identification of a promoter upstream of the immunity gene that is activated by antibiotic induction, that is, SOS box. Conclusions TDP analysis, coupled with other techniques (e.g., antibiotic induction, chemical reduction, plasmid sequencing, and in silico protein modeling), is a powerful tool to identify proteins (and their modifications), including prophage‐ and plasmid‐encoded proteins, produced by pathogenic microorganisms.

Biochemistry & Molecular Biology↗

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