Optimal Induction Research Articles

A Kernel-Free Boundary Integral Method for 2-D Magnetostatics Analysis

Performing magnetostatic analysis accurately and efficiently is very important for multi-objective optimization of electromagnetic device designs. In this research, a kernel-free boundary integral method (KFBIM) has been introduced for solving magnetic computations in a toroidal core geometry in 2-D. This study is very relevant in the design and optimization of toroidal inductors or transformers used in electrical systems, where lighter weight, higher inductance, higher efficiency, and lower leakage flux are required. The governing partial differential equations (PDEs) have been formulated as a system of the Fredholm integral equations of the second kind. Unlike traditional boundary integral methods or boundary element methods, KFBIM does not require an analytical form of Green’s function for evaluating integrals via numerical quadrature. Instead, KFBIM computes integrals by solving an equivalent interface problem on a Cartesian mesh. Compared with traditional finite difference methods for solving the governing PDEs directly, KFBIM produces a well-conditioned linear system. Therefore, the KFBIM requires only a fixed number of iterations when an iterative method [e.g., generalized minimal residual method (GMRES)] is applied for solving the linear system, and the numerical solution is not sensitive to computer round-off errors. The obtained results are then compared with a commercial finite element solver (ANSYS), which shows excellent agreement. It should be noted that, compared with FEM, the KFBIM does not require a body-fit mesh and can achieve high accuracy with a coarse mesh. In particular, the calculations of the magnetic potential and the tangential field intensity on the boundaries are more stable and exhibit almost no oscillations.

Additional Benefits of Rituximab and Plasma Exchange on Top of Standard Induction Therapy in Kidney Transplant Recipients With a Negative CDC Crossmatch but High Preformed Donor Specific Antibody Titer.

Optimal induction strategy in highly sensitized kidney transplant recipients (KTRs) is still a matter of debate. The place of therapies, such as plasma exchange and rituximab, with potential side effects and high cost, is not clearly established. We compared two induction strategies with (intensive) or without (standard) rituximab and plasma exchange in KTRs with high levels of preformed DSA transplanted between 2012 and 2019. Sixty KTRs with a mean age of 52.2 ± 12.2 years were included, 36 receiving standard and 24 intensive induction. Mean fluorescence intensity of immunodominant DSA in the cohort was 8,903 ± 5,469 pre-transplantation and similar in both groups. DSA level decrease was similar at 3 and 12months after transplantation in the two groups. An intensive induction strategy was not associated with better graft or patient survival, nor more infectious complications. The proportion of patients with rejection during the first year was similar (33% in each group), but rejection occurred later in the intensive group (211 ± 188 days, vs. 79 ± 158 days in the standard group, p < 0.01). Our study suggests that an intensive induction therapy including rituximab and plasma exchanges in highly sensitized kidney recipients is not associated with better graft survival but may delay biopsy-proven rejection.

A Multi-Enzyme Cascade for Efficient Production of Pyrrolidone from l-Glutamate.

Pyrrolidone is a high value-added monomer and an important active drug intermediate. However, the efficient enzymatic synthesis of pyrrolidone remains a challenge. Here, we developed and reconstructed a three-enzyme cascade pathway using Escherichia coli BL21(DE3) for the production of pyrrolidone from l-glutamate (l-Glu). The carnitine-CoA ligase from Escherichia coli (EcCaiC) at a low expression level and with a low activity is regarded as the rate-limiting enzyme. Here, we obtained the best EcCaiCF380M/N430D double mutant with a kcat/Km value 1.5 times higher than that of the wild type via mechanism-based protein engineering. For this, we (i) eliminated the steric hindrance of the loop ring to improve the precatalytic conformation of the adenylation intermediate and (ii) fixed the hinge region to stabilize the closed conformation of the enzyme. Furthermore, ribosome-binding site (RBS) optimization led to an increase in the expression level of EcCaiCF380M/N430D, which was then cloned into the plasmid pET-EcCaiCF380M/N430D-DegoPPK2. Finally, under optimal induction and transformation conditions, 16.62 g/L of pyrrolidone was generated from 30 g/L l-Glu (batch feeding) within 24 h with a molar conversion rate of 95.2% and the highest productivity ever obtained, to our knowledge (0.69 g/L/h). Our findings demonstrate a strategy that is potentially attractive for the industrial production of pyrrolidone. IMPORTANCE This study developed a three-enzyme cascade pathway for the production of pyrrolidone from l-Glu. The catalytic efficiency of carnitine CoA ligase from Escherichia coli (EcCaiC) was improved by mechanism-based protein engineering, and the titer of pyrrolidone was further increased by ribosome-binding site (RBS), induction conditions, and conversion conditions optimization. Finally, we efficiently produced pyrrolidone by one pot in vivo with 95.2% conversion and 0.69 g/L/h productivity. Our study provides a new possibility for the industrial production of enzymatic synthesis of pyrrolidone.

Whole-Cell Biocatalytic Production of Acetoin with an aldC-Overexpressing Lactococcus lactis Using Soybean as Substrate

Douchi is a traditional Chinese fermented soybean product, in which acetoin is a key flavor substance. Here, the α-acetolactate decarboxylase gene aldC was cloned from Lactiplantibacillus (L.) plantarum and overexpressed in Lactococcus (L.) lactis NZ9000 by nisin induction. The ALDC crude enzyme solution produced an enzyme activity of 35.16 mU. Next, whole cells of the recombinant strain NZ9000/pNZ8048-aldC were employed as the catalyst to produce acetoin in GM17 medium. An optimization experiment showed that an initial OD600 of 0.6, initial pH of 7.5, nisin concentration of 20 ng/mL, induction temperature of 37 °C and static induction for 8 h were the optimal induction conditions, generating the maximum acetoin production (106.93 mg/L). Finally, after incubation under the optimal induction conditions, NZ9000/pNZ8048-aldC was used for whole-cell biocatalytic acetoin production, using soybean as the substrate. The maximum acetoin yield was 79.43 mg/L. To our knowledge, this is the first study in which the aldC gene is overexpressed in L. lactis and whole cells of the recombinant L. lactis are used as a biocatalyst to produce acetoin in soybean. Thus, our study provides a theoretical basis for the preparation of fermented foods containing high levels of acetoin and the biosynthesis of acetoin in food materials.

The AMP-Activated Protein Kinase (AMPK) Positively Regulates Lysine Biosynthesis Induced by Citric Acid in Flammulina filiformis.

Flammulina filiformis, the most produced edible mushroom species in China, is rich in lysine. Further enhancing its lysine biosynthesis is vital for improving its quality in industrialized cultivation. Citric acid induction significantly increases both the biomass and growth rate of F. filiformis hyphae, as well as the lysine content. The genes encoding enzymes in the lysine biosynthesis pathway were detected under the optimal induction, revealing that the expression levels of hcs, hac, and hah were 2.67, 1.97, and 1.90 times greater, respectively, relative to the control, whereas no significant difference was seen for hdh, aat, sr, and shd, and the expression of aar decreased. Furthermore, the transcriptional levels of Ampk, GCN2, GCN4, and TOR were found significantly upregulated, with the most upregulated, Ampk, reaching a level 42.68 times greater than that of the control, while the phosphorylation of AMPK rose by nearly 54%. In AMPK-silencing strains under the optimal induction, however, the phosphorylation increment dropped to about 16% and the lysine content remained at the same level as in the WT. Thus, AMPK is presented as the critical intermediary in citric acid's regulation of lysine biosynthesis in F. filiformis.

In Vitro Propagation and Histochemical Analysis of Launaea sarmentosa (Willd.) Kuntze

Launaea sarmentosa (Willd.) Kuntze., an edible perennial herb, is critically considered an endangered species due to its habitat destruction. In vitro L. sarmentosa propagation was therefore conducted to preserve this plant. Two types of explants including leaf and stolon were applied to induce the multiple shoots by culturing on Murashige and Skoog (MS) medium supplemented with BA (0, 1, and 2 mg/L) and 2,4-D (0, 1, 2, and 4 mg/L). The optimal shoot induction medium for both explants was MS medium containing 2 mg/l BA (presented 100 % shoot induction). Roots were induced by various types and concentrations of auxins [1-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA)]. The highest percentage of rooting (70.00 ± 11.05 %) was obtained from stolon-derived shoots on MS medium supplemented with 0.1 mg/l IAA. One-hundred percent survivability was presented after transforming plantlets into a mixture of soil, sand, and peat moss (1:1:1). Starch, protein, and lipid were accumulated in leaves detected by histochemical technique. The obtained information will be a useful tool for mass propagation of L. sarmentosa and to obtain basic knowledge for plant accumulating substances. HIGHLIGHTS This paper highlights 2 types of sarmentosa explant including leaves and stolons have great potential to produce multiple shoots after inoculation in MS medium adding BA. All regenerated plants get success for acclimatization and transplantation to the greenhouse. In vitro plantlets of L. sarmentosa leaves accumulated fat, protein and carbohydrate which might be related with precursor of medicinal properties GRAPHICAL ABSTRACT

Participation of ATM, SMG1, and DDX5 in a DNA Damage-Induced Alternative Splicing Pathway.

Altered cellular responses to DNA damage can contribute to cancer development, progression, and therapeutic resistance. Mutations in key DNA damage response factors occur across many cancer types, and the DNA damage-responsive gene, TP53, is frequently mutated in a high percentage of cancers. We recently reported that an alternative splicing pathway induced by DNA damage regulates alternative splicing of TP53 RNA and further modulates cellular stress responses. Through damage-induced inhibition of the SMG1 kinase, TP53 pre-mRNA is alternatively spliced to generate TP53b mRNA and p53b protein is required for optimal induction of cellular senescence after ionizing radiation-induced DNA damage. Herein, we confirmed and extended these observations by demonstrating that the ATM protein kinase is required for repression of SMG1 kinase activity after ionizing radiation. We found that the RNA helicase and splicing factor, DDX5, interacts with SMG1, is required for alternative splicing of TP53 pre-mRNA to TP53b and TP53c mRNAs after DNA damage, and contributes to radiation-induced cellular senescence. Interestingly, the role of SMG1 in alternative splicing of p53 appears to be distinguishable from its role in regulating nonsense-mediated RNA decay. Thus, ATM, SMG1, and DDX5 participate in a DNA damage-induced alternative splicing pathway that regulates TP53 splicing and modulates radiation-induced cellular senescence.

Preparation of IgE Antibody and Distribution of IgE+ Secretory Cells in the Palatine Tonsil of Bactrian Camel.

Allergic diseases induced by dust have seriously threatened human health, while Bactrian camels can live in a sandy environment for a long time. To prepare rabbit anti-Bactrian camel IgE antibody and explore the distribution characteristics of IgE+ secretory cells in the palatine tonsils, which lays a theoretical foundation for the distribution of local antibodies in the palatal tonsils of Bactrian camel and the study of immune function. In this study, the amino acid sequences of Bactrian camel IgE, IgA, IgM and IgG heavy chain constant regions were compared, and a specific IgE gene fragment were selected (447 bp). The recombinant plasmid pET-28a-IgE was induced in Escherichia coli BL21(DE3) by IPTG and its expression conditions were optimized. The antibody was prepared by immunizing rabbits with purified IgE recombinant protein, its titer and specificity were detected by indirect ELISA and Western blotting. Immunohistochemical and statistical methods investigated the distribution of IgE+ secretory cells in the palatine tonsils. The IgE recombinant protein was expressed in the form of inclusion bodies with a size of 16 kDa. The optimal IPTG induction concentration was 0.7 mmol/L and the induction time was 8 h. The titer of the antibody was 1:16000 by ELISA, and the antibody could specifically bind to the recombinant protein by Western blotting. IgE+ secretory cells were mainly distributed in the subepithelial compartments of reticulated crypt epithelium of the palatine tonsil of the Bactrian camel, followed by the subepithelial compartments of stratified squamous epithelium and occasionally in the extrafollicular region. The rabbit anti-Bactrian camel IgE polyclonal antibody was successfully prepared. It is confirmed that IgE exists in the palatine tonsils of Bactrian camels under normal living conditions. In addition, IgE+ secretory cells are mainly distributed in the subepithelial compartments of reticulated crypt epithelium of the palatine tonsil, which is consistent with the distribution characteristics of IgG+ and sIgA+ secretory cells in the palatal tonsils of the Bactrian camel.

Culture of leukocyte-derived cells from human peripheral blood: Increased expression of pluripotent genes OCT4, NANOG, SOX2, self-renewal gene TERT and plasticity.

There are few stem cells in human peripheral blood (PB). Increasing the population and plasticity of stem cells in PB and applying it to regenerative medicine require suitable culture methods. In this study, leukocyte populations 250 mL of PB were collected using a blood separator before that were cultured in optimal cell culture medium for 4 to 7 days. After culturing, stemness characteristics were analyzed, and red blood cells were removed from the cultured cells. In our results, stemness markers of the leukocyte populations Sca-1+ CD45+, CD117+ CD45+, and very small embryonic-like stem cells CD34+ Lin- CD45- and CXCR4+ Lin- CD45- were significantly increased. Furthermore, the expression of stem cell genes OCT4 (POU5F1), NANOG, SOX2, and the self-renewal gene TERT was analyzed by quantitative real-time polymerase chain reaction in these cells, and it showed a significant increase. These cells could be candidates for multi-potential cells and were further induced using trans-differentiation culture methods. These cells showed multiple differentiation potentials for osteocytes, nerve cells, cardiomyocytes, and hepatocytes. These results indicate that appropriate culture methods can be applied to increase expression of pluripotent genes and plasticity. Leukocytes of human PB can be induced to trans-differentiate into pluripotent potential cells, which will be an important breakthrough in regenerative medicine.

Multilevel Dual Active Bridge Leakage Inductance Selection for Various DC-Link Voltage Spans

The leakage inductance of the transformer in a dual active bridge (DAB) dc–dc converter directly impacts the ac current waveforms and the power factor; thus, it can be considered a design requirement for the transformer. In the existing literature, a choice is made to either ensure soft switching in nominal power or to minimize the RMS current of the transformer. The inductance is typically obtained using optimization procedures. Implementing these optimizations is time-consuming, which can be avoided if a closed-form equation is derived for the optimum leakage inductance. In this paper, analytical formulas are derived to estimate the desired leakage inductance such that the highest RMS value of the current in the operation region of a DAB is kept to its minimum value. The accuracy and sensitivity of the analytical solutions are evaluated. It is shown that in a large design domain, the solution for the YY-connected MFT has a less than 3% error compared to the results obtained from an optimization engine. As an example of the importance of selecting the leakage inductance correctly, it is shown that for 11% deviations in the dc link voltages, a 10% deviation from the desired leakage inductance value can cause 2% higher RMS currents in the converter.

Design, Analysis and Optimization of Magnetic-Core Solenoid Inductor for On-Chip Multi-Phase Buck Converter

Due to heterogeneous integration, there is an increased demand for on-chip DC-DC converters to provide multiple voltage domains for efficient operation of the system. Towards this, planar magnetic-core based inductors gained popularity due to their increased inductance density for on-chip buck converter realization. To ensure proper operation of the converter, the magnetic-core should not saturate, and over-design of the inductor decreases the converter efficiency significantly. Also, the magnetic-core area budget is limited for on-chip design. This paper proposes an optimization framework to design an efficient multi-phase buck converter with the magnetic-core solenoid inductor for the given specifications. We investigate two cost-functions for optimization where Cost function <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">1</sub> considers only power loss reduction, whereas Cost function <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> considers both power loss and inductor footprint. Full-wave simulations on commercial tool show that the optimized multi-phase buck converter with 500 mA load current achieves 88.13% and 88.06% efficiency for Cost function <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">1</sub> and Cost function <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> , respectively. The footprint for the optimized inductor with Cost function <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> has a 10% reduction compared with the optimized inductor with Cost function <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">1</sub> .

Optimal lameness induction model development using amphotericin B in meat goats.

Lameness continues to be a critical health and welfare concern associated with goat production. Amphotericin B (amp B) is an antimicrobial successful in inducing transient lameness for research purposes previously in livestock animals. The objectives of this study were to (1) identify which of three varying doses of amp B would be most effective in inducing lameness in meat type goats and (2) develop a facial grimace scale for goats. Lameness was produced by an intra-articular injection of amphotericin B into the left hind lateral claw distal interphalangeal joint with either a 5 mg/0.25 mL (high-low, 5 mg of amphotericin B in a volume of 0.25 mL), 5 mg/0.5 mL (high-high, 5 mg of amphotericin B in a volume of 0.5 mL), or a 2.5 mg/0.25 mL (low-low, 2.5 mg of amphotericin B in a volume of 0.25 mL). A saline treatment of 0.5 mL was used as control (0.9% sterile saline solution). Lameness response was analyzed by infrared thermography (IRT) at the induced joint, mechanical-nociception threshold (MNT), visual lameness scoring (VLS), a visual analogue scale (VAS), kinetic gait analysis (KGA), plasma cortisol (CORT), substance P (Sub P), and behavior scoring. The IRT and MNT values differed by timepoint (P ≤ 0.0001). Results from VLS showed the HL treatment was the most effective at inducing lameness (6/6 goats became lame compared to HH 4/6 and LL 2/6). At 24, 48, and 72 h, VAS scores were significantly higher when comparing HL to all other treatment groups (P = 0.0003). Both behavior observers (1 and 2) reported a significant time effect (P = 0.05), with goats exhibiting more facial grimacing at 24 h post-lameness induction. From these data, an optimal dose for a repeatable lameness induction model in goats was aquired. An effective Goat Grimace Scale (GGS) was also developed to evaluate pain responses in goats.

Assessment of sol-gel derived iron oxide substituted 45S5 bioglass-ceramics for biomedical applications.

Magnetic bioactive glass-ceramic (MGC) powders with nominal compositions of (45 - x)SiO224.5CaO24.5Na2O6P2O5xFe2O3 (x = 2, 4, 6, 8, 10, and 15 wt%) have been synthesized by a sol-gel route by systematically substituting silicon dioxide with iron oxide in Hench's 45S5 glass composition. Powder X-ray diffraction studies revealed a variation in the percentage of combeite (Ca2Na2Si3O9), magnetite (Fe3O4), and hematite (Fe2O3) nanocrystalline phases in MGC powders as a function of composition. Zeta potential measurements showed that MGC containing up to 10 wt% iron oxide formed stable suspensions. The saturation magnetization and heat generation capacity of MGC fluids increased with an increase in iron oxide content. Degradation of MGC powders was investigated in phosphate buffered saline (PBS). The in vitro bioactivity of the MGC powders taken in pellet form was confirmed by observing the pH variation as well as hydroxyapatite layer (HAp) formation upon soaking in modified simulated body fluid. These studies showed a decrement in the overall bioactivity in samples with high iron oxide content due to the proportional decrease in the silanol group. Monitoring the proliferation of MG-63 osteoblast cells in Dulbecco's Modified Eagle Medium (DMEM) revealed that MGC with up to 10 wt% iron oxide exhibited acceptable viability. The systematic study revealed that the MGC with 10 wt% iron oxide exhibited optimal cell viability, magnetic properties and induction heating capacity, which were better than those of FluidMag-CT, which is used for hyperthermia treatment.

C5aR antagonist inhibits LPS-induced inflammation in human gingival fibroblasts via NF-κB and MAPK signaling pathways.

Abnormal complement activation is associated with periodontitis. W54011 is a novel non-peptide C5aR antagonist (C5aRA) that exhibits favorable anti-inflammatory effects in various inflammatory models. However, whether W54011 inhibits periodontitis has not yet been fully elucidated. To address this, we have investigated the probable anti-inflammatory mechanism of W54011 in LPS-treated inflammation in human gingival fibroblasts (HGFs). HGFs were isolated from healthy gingival tissue samples using the tissue block method and were identified with immunofluorescence staining. The CCK8 assay and reverse transcription-PCR (RT-PCR) were used to select the optimal induction conditions for Lipopolysaccharide (LPS) and C5aRA (according to supplementary data S1, S2 and S3). The levels of inflammatory cytokines, C5aR, and the activation of NF-κB/MAPK signaling pathways were determined by RT-quantitative PCR (RT-qPCR) and Western blotting. Immunofluorescence results showed that vimentin and FSP-1 were positive in HGFs and Keratin was negative in HGFs. Immunofluorescence staining demonstrated that C5aRA inhibited LPS-stimulated nuclear translocation of p-p65. RT-qPCR and Western blotting showed that C5aRA reduced the expression of IL-1β, IL-6, TNF-α, C5aR, p-p65, p-IκBα, p-JNK, p-c-JUN, and TLR4 in LPS-induced HGFs. These findings suggested that C5aRA attenuated the release of inflammatory cytokines in LPS-induced HGFs by blocking the activation of the NF-κB and MAPK signaling pathways.

How to design optimal induction pathways for pharmacy professionals

How to design optimal induction pathways for pharmacy professionals

Novel active-passive hybrid piezoelectric network for vibration suppression in fluid-conveying pipes

Given the failures like fatigue and fracture of the pipes conveying fluid induced by undesired dynamics and instability and the traditional passive damping cannot effectively attenuate the vibration in the low-frequency bands, this work introduces a novel active-passive control technique, namely, the hybrid piezoelectric network consisting of the piezoelectric layer, the circuit of the resistance and inductance, voltage source, controller, and sensor, which is attached on the cantilevered fluid-conveying pipes to suppress the vibration in wider frequency-bands. Built upon the location of the neutral axis in the segment of pipes is varied due to the integration of the pipeline and piezoelectric actuator, the governing equations of the fluid-conveying pipes attached with the hybrid piezoelectric network are established by virtue of Hamilton's principle in conjunction with the assumed mode method, and approximate analytical expressions of frequency responses are derived using the Laplace transform technique. The passive damping performance is carried out to achieve insightfully using the open-loop analysis based on the circuit of the resistance and inductance is set as the passive shunt characteristic, then the closed-loop features are also investigated according to the velocity feedback control. The numerical example illustrates the presented control method is effective and feasible to realize the vibration suppression on the pipes conveying fluid. Based on this, the optimal resistance and inductance are proposed through the theoretical analysis, which can further broaden the vibration attenuation.

Characterization of a Stress-Enhanced Promoter from the Grass Halophyte, Spartina alterniflora L.

Stress-inducible promoters are vital for the desirable expression of genes, especially transcription factors, which could otherwise compromise growth and development when constitutively overexpressed in plants. Here, we report on the characterization of the promoter region of a stress-responsive gene SaAsr1 from monocot halophyte cordgrass (Spartina alterniflora). Several cis-acting elements, such as ABRE (ABA-responsive element), DRE-CRT (dehydration responsive-element/C-Repeat), LTRE (low temperature-responsive element), ERE (ethylene-responsive element), LRE (light-responsive element), etc. contributed at varying degrees to salt-, drought- and ABA-enhanced expression of gusA reporter gene in Arabidopsis&amp;nbsp;thaliana under the full-length promoter, pAsr11875 and its deletion derivatives with an assortment of cis-regulatory motifs. The smallest promoter, pAsr1491, with three cis-acting elements (a CCAAT box-heat responsive, an LRE, and a copper responsive element) conferred drought-enhanced expression of gusA; pAsr1755 (with an ABRE and a DRE) presented the highest expression in ABA and drought; and pAsr1994 with seven ABREs and two DREs conferred optimal induction of gusA, especially under drought and ABA. Arabidopsis transgenics expressing a known abiotic stress-responsive gene, SaADF2 (actin depolymerization factor 2), under both pAsr11875 and p35S promoters outperformed the wild type (WT) with enhanced drought and salt tolerance contributed by higher relative water content and membrane stability with no significant difference between pAsr11875:SaADF2 or p35S:SaADF2 lines. However, pAsr11875:SaADF2 lines produced healthy plants with robust shoot systems under salt stress and control compared to slightly stunted growth of the p35S:SaADF2 plants. This reestablished the evidence that transgene expression under a stress-inducible promoter is a better strategy for the genetic manipulation of crops.

Active Immunization Against NMDA NR1 Subunit as a Model of Autoimmune Encephalitis

Objective To identify and develop the optimal active immunization induction method for NMDAR encephalitis in rodents. Background Encephalitis is a devastating neurologic disorder with high morbidity and mortality. Many cases are autoimmune. N-Methyl-D-aspartic acid receptor (NMDAR) encephalitis (NMDARE), characterized by antibodies against the NMDAR in the blood and spinal fluid of patients, is the most common form of autoimmune encephalitis (AE). A translational rodent model of NMDARE would allow for in-depth studies into AE pathophysiology, leading to advances in the diagnosis and treatment of this debilitating neuropsychiatric disorder. Design/Methods 7-week-old female C57BL/6J mice were injected subcutaneously with an emulsion of complete Freund's adjuvant, attenuated Mycobacterium tuberculosis (TB), and a 30 amino acid peptide flanking the NMDAR NR1 subunit N368/G369 residue targeted by antibodies in NMDARE patients. Three different induction methods were tested by varying the amount and injection method of pertussis toxin, subcutaneous injection sites, reimmunization, and amounts of TB. Mice were bled biweekly and sacrificed at 2, 4, 6, 8, and 14 weeks. Serum and CSF NMDAR antibody titer; mouse behavior; hippocampal NMDAR protein and cluster density; and brain immune cell entry and cytokine content were examined. Results Immunized mice had serum and CSF NMDAR antibodies. Mice exhibited behavioral changes, altered hippocampal NMDAR protein, brain immune cell entry, and elevated cytokines in their brains. Titers were higher and changes were sustained in reimmunized mice. Conclusions Active immunization against the portion of the NMDAR targeted in patients with NMDARE resulted in robust production of NMDAR antibodies in the blood and spinal fluid, changes in hippocampal NMDAR protein, elevations in brain immune cells and cytokines, and behavioral changes in mice. Reimmunization was needed to sustain the responses. Active immunization therefore holds potential as a translational model of NMDARE, allowing for the creation of a novel generation of diagnostics and therapeutics.

Transcriptome and metabolome analyses reveal the efficiency of in vitro regeneration by TDZ pretreatment in mulberry

Thidiazuron (TDZ) is a plant growth regulator that is widely used as a defoliant and cytokinin-like compound. Additionally, mulberry is an economically important crop. However, regeneration is difficult in most mulberry species, and the regulatory mechanism controlling the regeneration process remains unclear. In this work, an efficient in vitro shoot regeneration system for mulberry was established by TDZ application. The results showed that the optimum adventitious bud induction, 70.83% in tetraploid M. notabilis cv. Chuanbei1 (CB1) and 96.82% in M. abla cv. Yunhua (YH), was obtained by the shoot pretreatment method using petiolate leaves as explants. The high-frequency regeneration petiolate leaf explants of YH treated with TDZ (T8) and the control (CK) were subjected to multiomic analyses. The comparison of the TDZ-treated with the CK groups identified 494 upregulated differentially expressed genes (DEGs) and 293 downregulated DEGs. GO analysis indicated that these DEGs were related mostly to integral component of membrane, transcription factor activity, and phosphorelay signal transduction system. Metabolomic analysis identified 1,889 compounds, including 205 differentially accumulated metabolites (DAMs). Of these DAMs, the number of lipids and lipid-like molecules was highest and accounted for 27.8% of all DAMs. Transcriptome profile, metabolomic, and phytohormone integrated analyses revealed that TDZ rearranged adenine-type cytokinins (CTKs) and negatively regulated endogenous CTK signal transduction. Moreover, TDZ treatment enhanced auxin signal transduction by increasing the expression of auxin flux carrier genes and downstream regulated genes without changing auxin biosynthesis in mulberry. In summary, this study provides a new method for mulberry in vitro regeneration and contributes to a better understanding of the transcriptional and metabolic changes that occur in mulberry after TDZ treatment.

Increased Light Olefin Production by Sequential Dehydrogenation and Cracking Reactions

In this study, a sequential reaction using selected metal oxides, followed by ZSM-5-based catalysts, was employed to demonstrate a promising route for enhancing light olefin production in the catalytic cracking of naphtha. The rationale for the reaction is based on the induction of alkenes into hydrocarbon feeds prior to cracking. The optimum olefin induction was achieved by carefully optimizing the dehydrogenation active sites Mo/Al2O3 catalyst. The formed alkenes have a lower activation energy for C-H/C-C bond breaking compared to alkanes. This could accelerate the formation of carbenium ions, thus promoting the conversion of n-octane to produce light olefins. Detailed product distribution and DFT calculation indicated a remarkable increase in ethylene and propylene production in the final product through a modified reaction pathway. Compared with the common metal-promoted zeolite catalysts, the new route could avoid the block of zeolite channels and corresponding decreased catalytic cracking activity. The feasibility of the proposed route was confirmed with different ratios of dehydrogenation catalyst to the reactant. The highest yields of ethylene and propylene reached 13.22% and 33.12% with ratios of Mo/Al2O3 and ZSM-5-based catalyst to n-octane both 10:1 at 600 °C. Stability tests showed that the catalytic activity of the double-bed system was stable over 10 cycles.