Sublethal effect and detoxifying metabolism of metaflumizone and indoxacarb on the fall armyworm, Spodoptera frugiperda.

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) (Lepidoptera, Noctuidae), is a highly polyphagous invasive pest that damages various crops. Pesticide control is the most common and effective strategy to control FAW. In this study, we evaluated the toxicity of metaflumizone and indoxacarb against third-instar FAW larvae using the insecticide-incorporated artificial diet method under laboratory conditions. Both metaflumizone and indoxacarb exhibited substantial toxicity against FAW, with LC50 values of 2.43 and 14.66 mg/L at 72 h, respectively. The sublethal effects of metaflumizone and indoxacarb on parental and F1 generation FAW were investigated by exposing third-instar larvae to LC10 and LC30 concentrations of these insecticides. Sublethal exposure to these two insecticides significantly shortened adult longevity, extended pupal developmental times and led to reduced pupal weight, pupation rates, and adult fecundity in the treated parental generation and F1 generation at LC10 or LC30 concentrations, in comparison to the control group. The larval developmental times were shortened in the parental generation but prolonged in the F1 generation, after being treated with sublethal concentrations of metaflumizone. Furthermore, larvae exposed to LC10 or LC30 concentrations of indoxacarb exhibited elevated activity levels of cytochrome P450 monooxygenase and glutathione S-transferase, which coincides with the observed synergistic effect of piperonyl butoxide and diethyl maleate. In conclusion, the high toxicity and negative impact of metaflumizone and indoxacarb on FAW provided significant implications for the rational utilization of insecticides against this pest.

Knockdown of the glutamate-gated chloride channel gene decreases emamectin benzoate susceptibility in the fall armyworm, Spodoptera frugiperda.

Emamectin benzoate (EB), a derivative of avermectin, is the primary insecticide used to control the fall armyworm (FAW) in China. However, the specific molecular targets of EB against FAW remain unclear. In this study, we cloned the glutamate-gated chloride channel (GluCl) gene, which is known to be a primary molecular target for avermectin. We first investigated the transcript levels of SfGluCl in FAW and found that the expression level of SfGluCl in the head and nerve cord was significantly higher than that in other tissues. Furthermore, we found that the expression level of SfGluCl was significantly higher in eggs than that in other developmental stages, including larvae, pupae, and adults. Additionally, we identified three variable splice forms of SfGluCl in exons 3 and 9 and found that their splice frequencies remained unaffected by treatment with the LC50 of EB. RNAi mediated knockdown of SfGluCl showed a significant reduction of 42% and 65% after 48 and 72 h of dsRNA feeding, respectively. Importantly, knockdown of SfGluCl sifgnificantly reduced LC50 and LC90 EB treatment induced mortality of FAW larvae by 15% and 44%, respectively, compared to the control group feeding by dsEGFP. In contrast, there were no significant changes in the mortality of FAW larvae treated with the control insecticides chlorantraniliprole and spinetoram. Finally, molecular docking simulations revealed that EB bound to the large amino-terminal extracellular domain of SfGluCl by forming five hydrogen bonds, two alkyl hydrophobic interactions and one salt bridge. These findings strongly suggest that GluCl may serve as one of the molecular targets of EB in FAW, shedding light on the mode of action of this important insecticide.

A Performance Improvement for Indoor Positioning Systems Using Earth's Magnetic Field.

Although most indoor positioning systems use radio waves, such as Wi-Fi, Bluetooth, or RFID, for application in department stores, exhibition halls, stations, and airports, the accuracy of such technology is easily affected by human shadowing and multipath propagation delay. This study combines the earth's magnetic field strength and Wi-Fi signals to obtain the indoor positioning information with high availability. Wi-Fi signals are first used to identify the user's area under several kinds of environment partitioning methods. Then, the signal pattern comparison is used for positioning calculations using the strength change in the earth's magnetic field among the east-west, north-south, and vertical directions at indoor area. Finally, the k-nearest neighbors (KNN) method and fingerprinting algorithm are used to calculate the fine-grained indoor positioning information. The experiment results show that the average positioning error is 0.57 m in 12-area partitioning, which is almost a 90% improvement in relation to that of one area partitioning. This study also considers the positioning error if the device is held at different angles by hand. A rotation matrix is used to convert the magnetic sensor coordinates from a mobile phone related coordinates into the geographic coordinates. The average positioning error is decreased by 68%, compared to the original coordinates in 12-area partitioning with a 30-degree pitch. In the offline procedure, only the northern direction data are used, which is reduced by 75%, to give an average positioning error of 1.38 m. If the number of reference points is collected every 2 m for reducing 50% of the database requirement, the average positioning error is 1.77 m.

Elucidating the Associated Biological Function and Clinical Significance of RHOJ Expression in Urothelial Carcinoma.

Urothelial cancer, a common urinary system malignancy, often presents treatment challenges due to metastasis and chemotherapy side effects. Angiogenesis, crucial for tumor growth, has become a target for drug development. This study explores the expression, prognostic value, and clinical correlation of RHOJ in the TCGA BLCA, GSE31684, and GSE32894 datasets. We identify common differentially expressed genes across these databases and utilize g:Profiler and Cytoscape ClueGO for functional assessment. Further, we perform a gene set enrichment analysis (GSEA) using Hallmark gene sets and use the imsig package for immune cell infiltration analysis. Our analysis indicates that RHOJ expression levels significantly impact survival rates, tumor progression, and immune response in urothelial tumors. High RHOJ expression correlated with poor prognosis, advanced disease stages, and an increase in monocyte population within the tumor microenvironment. This aligns with current literature indicating a key role of immune infiltration in bladder cancer progression and treatment response. Moreover, the GSEA and imsig results further suggest a potential mechanistic link between RHOJ expression and immune-related pathways. Considering the increasing emphasis on immunotherapeutic strategies in bladder cancer management, our findings on RHOJ's potential as a diagnostic biomarker and its association with immune response open new avenues for therapeutic interventions.

Cu-Catalyzed Asymmetric Kinetic Boron Conjugate Addition of γ-Substituted α,ß-Unsaturated γ-Lactams.

Chiral α,ß-unsaturated γ-lactams bearing simple γ- substituents are found in biologically active molecules and natural products, however, their synthesis still remains difficult. Herein, we report an efficient kinetic resolution (KR) of γ-substituted α,ß-unsaturated γ-lactams via a Cu-catalyzed asymmetric boron conjugate addition, which also leads to the efficient synthesis of chiral ß-hydroxy-γ-lactams with ß,γ-stereogenic carbon centers. The KR proceeded smoothly with a wide range of γ-alkyl or aryl substituted substrates including those bearing aromatic heterocycles and different N-protected substrates in up to 347 of s value. Their highly versatile transformations, synthetic utility in biologically active molecules, and inhibitory activities against cisplatin-sensitive ovarian cancer cell A2780 have also been demonstrated. Differing from the well-known mechanism involving Cu-B species in Cu-catalyzed boron conjugate additions, our mechanistic studies using density functional theory (DFT) calculations and experiments indicate that a Lewis acid CuI -catalyzed mechanism is the likely pathway in the catalytic reaction.

Enantio- and Diastereoselective Synthesis of Chiral Tetrasubstituted α-Amino Allenoates Bearing a Vicinal All-Carbon Quaternary Stereocenter with Dual-Copper-Catalysis.

The skeletons of chiral tetrasubstituted allenes bearing a vicinal all-carbon quaternary stereocenter are of importance but still challenging to synthesize. Herein, we report enantio- and diastereoselective γ-additions of 1-alkynyl ketimines with dual-copper-catalysis under mild conditions, affording chiral tetrasubstituted α-amino allenoates bearing a vicinal all-carbon quaternary stereocenter in high yields (up to 99 % yield) with excellent enantioselectivities (up to 99 % ee) and diastereoselectivities (up to >20 : 1 dr). Importantly, the stereodivergent synthesis of the products was realized by the asymmetric γ-addition reaction and the Grignard reagent promoted epimerization. Moreover, the dual-copper-catalyzed γ-addition reactions were smoothly applied to a gram-scale reaction and adopted to introduce chiral tetrasubstituted allenyl moieties into bioactive molecules. Mechanistic experiments and density functional theory (DFT) calculations demonstrated that the asymmetric γ-addition reactions were catalyzed by double chiral copper catalysts.

Defect-Free Arbitrary-Geometry Assembly of Mixed-Species Atom Arrays.

Optically trapped mixed-species single atom arrays with arbitrary geometry are an attractive and promising platform for various applications, because tunable quantum systems with multiple components provide extra degrees of freedom for experimental control. Here, we report the first demonstration of two-dimensional 6×4 dual-species atom assembly of ^{85}Rb (^{87}Rb) atoms with a filling fraction of 0.88 (0.89). This mixed-species atomic synthesis is achieved via rearranging initially randomly distributed atoms by a sorting algorithm (heuristic heteronuclear algorithm) which is designed for bottom-up atom assembly with both user-defined geometries and two-species atom number ratios. Our fully tunable hybrid-atom systems with scalable advantages are a good starting point for high-fidelity quantum logic, many-body quantum simulation, and single molecule array formation.

Copper-Catalyzed Regioselective [3+3] Annulations of Alkynyl Ketimines with α-Cyano Ketones: the Synthesis of Polysubstituted 4H-Pyran Derivatives with a CF3 -Containing Quaternary Center.

Regioselective [3+3] annulation of alkynyl ketimines with α-cyano ketones for the synthesis of polysubstituted 4H-pyran derivatives with a quaternary CF3 -containing center has been realized by using Cu(OAc)2 as the catalyst. The novel strategy tolerates a wide range of α-CF3 alkynyl ketimines and α-cyano ketones with both aryl and alkyl substitutents. A preliminary asymmetric synthesis of chiral product 3 has been attempted by using copper and chiral thiourea as the cocatalyst with excellent yields (86-99 %) and good enantioselectivities (71-78 % ee). Furthermore, product 3 aa could be obtained on a gram-scale reaction with 75 % yield and 99 % ee after recrystallization. Several products were also transformed readily. Control experiments indicate that the reaction involves a process with a base-catalyzed or chiral thiourea-catalyzed Mannich-type reaction followed by a highly regioselective copper-catalyzed ring-closing reaction on the alkynyl moiety in a 6-endo-dig fashion.

Research on Chinese Speech Emotion Recognition Based on Deep Neural Network and Acoustic Features.

In recent years, the use of Artificial Intelligence for emotion recognition has attracted much attention. The industrial applicability of emotion recognition is quite comprehensive and has good development potential. This research uses voice emotion recognition technology to apply it to Chinese speech emotion recognition. The main purpose of this research is to transform gradually popularized smart home voice assistants or AI system service robots from a touch-sensitive interface to a voice operation. This research proposed a specifically designed Deep Neural Network (DNN) model to develop a Chinese speech emotion recognition system. In this research, 29 acoustic characteristics in acoustic theory are used as the training attributes of the proposed model. This research also proposes a variety of audio adjustment methods to amplify datasets and enhance training accuracy, including waveform adjustment, pitch adjustment, and pre-emphasize. This study achieved an average emotion recognition accuracy of 88.9% in the CASIA Chinese sentiment corpus. The results show that the deep learning model and audio adjustment method proposed in this study can effectively identify the emotions of Chinese short sentences and can be applied to Chinese voice assistants or integrated with other dialogue applications.

Cost-Effective Fitting Model for Indoor Positioning Systems Based on Bluetooth Low Energy.

Bluetooth Low Energy (BLE) is a positioning technology that is commonly used in indoor positioning systems (IPS) such as shopping malls or underground parking lots, because of its low power consumption and the low cost of Bluetooth devices. It also maintains high positioning accuracy. Since the cost of BLE itself is low, it has now been used in larger environments such as parking lots or shopping malls for a long time. However, it is necessary to configure a large number of devices in the environment to obtain accurate positioning results. The most accurate method of using signal strength for positioning is the signal pattern-matching method. The positioning result is compared through a database with the overheads of time and labor costs, since the amount of data will be proportional to the size of the environment for BLE-IPS. A planar model that conforms to the signal strength in the environment was generated, wherein the database comparison method is replaced by an equation solution, to improve various costs but diminish the positioning accuracy. In this paper, we propose to further replace the planar model with a cost-effective fitting model to both save costs and improve positioning accuracy. The experimental results demonstrate that this model can effectively reduce the average positioning error in distance by 31%.

Confocal laser endomicroscopy under propofol-based sedation for early gastric cancer and pre-cancerous lesions is associated with better diagnostic accuracy: a retrospective cohort study in China.

BACKGROUND: Confocal laser endomicroscopy (CLE) has advantages in detecting gastric neoplastic lesions, meanwhile it requires strict patient cooperation. Sedation could improve patient cooperation and quality of endoscopy. However, sedation is still not very popular in some resource-limited countries and regions. The purpose of this study was to compare propofol-based sedated versus un-sedated CLE in the value of diagnosing early gastric cancer (EGC) and precancerous lesions. METHODS: A retrospective, cohort, single center study of 226 patients who underwent CLE between January 1, 2015 and December 31, 2017 was performed. Patients enrolled were allocated into the propofol-based sedated group (n = 126) and the un-sedated group (n = 100). The comparison of validity and reliability of CLE for identifying EGC and precancerous lesions between the two groups was performed through analyzing CLE diagnosis and pathological diagnosis. Reporting followed the STROBE guidelines. RESULTS: The area under receiver operating characteristic curve (AUROC) of diagnosing EGC in the sedated group was 0.97 (95 % CI: 0.95 to 0.99), which was higher than that in the un-sedated group (0.88 (95 % CI: 0.80 to 0.97), P = 0.0407). CLE with sedation performed better than without sedation in diagnosing intraepithelial neoplasia and intestinal metaplasia (P = 0.0008 and P = 0.0001, respectively). For patients considered as high-grade intraepithelial neoplasia or EGC by endoscopists, they would not get biopsy during CLE but receive endoscopic submucosal dissection (ESD) subsequently, and the misdiagnosis rate of CLE was 0 % in the sedated group and 27.59 % (95 % CI: 10.30-44.91 %) in the un-sedated group (P = 0.006). CONCLUSIONS: Propofol based sedation was associated with improved diagnostic value of CLE for detecting EGC as well as precancerous lesions (intraepithelial neoplasia OR intestinal metaplasia).

Balanced Coherence Times of Atomic Qubits of Different Species in a Dual 3×3 Magic-Intensity Optical Dipole Trap Array.

We construct a polarization-mediated magic-intensity (MI) optical dipole trap (ODT) array, in which the detrimental effects of light shifts on the mixed-species qubits are efficiently mitigated so that the coherence times of the mixed-species qubits are both substantially enhanced and balanced for the first time. This mixed-species magic trapping technique relies on the tunability of the coefficient of the third-order cross term and ground state hyperpolarizability, which are inherently dependent on the degree of circular polarization of the trapping laser. Experimentally, polarization of the ODT array for ^{85}Rb qubits is finely adjusted to a definite value so that its working magnetic field required for magic trapping amounts to the one required for magically trapping ^{87}Rb qubits in another ODT array with fully circular polarization. Ultimately, in such a polarization-mediated MI-ODT array, the coherence times of ^{87}Rb and ^{85}Rb qubits are, respectively, enhanced up to 891±47 ms and 943±35 ms. Moreover, we reveal that the noise of the elliptic polarization causes dephasing effect on the ^{85}Rb qubits but it could be efficiently mitigated by choosing the dynamical range of active polarization device. We also show that light shifts seen by qubits in an elliptically polarized MI-ODT can be more efficiently compensated due to the decrease in the ground state hyperpolarizability. It is anticipated that the novel mixed-species MI-ODT array is a versatile platform for building scalable quantum computers with neutral atoms.

Expression system for structural and functional studies of human glycosylation enzymes.

Vertebrate glycoproteins and glycolipids are synthesized in complex biosynthetic pathways localized predominantly within membrane compartments of the secretory pathway. The enzymes that catalyze these reactions are exquisitely specific, yet few have been extensively characterized because of challenges associated with their recombinant expression as functional products. We used a modular approach to create an expression vector library encoding all known human glycosyltransferases, glycoside hydrolases, and sulfotransferases, as well as other glycan-modifying enzymes. We then expressed the enzymes as secreted catalytic domain fusion proteins in mammalian and insect cell hosts, purified and characterized a subset of the enzymes, and determined the structure of one enzyme, the sialyltransferase ST6GalNAcII. Many enzymes were produced at high yields and at similar levels in both hosts, but individual protein expression levels varied widely. This expression vector library will be a transformative resource for recombinant enzyme production, broadly enabling structure-function studies and expanding applications of these enzymes in glycochemistry and glycobiology.

The application of 595-nm pulsed dye laser for vascular anomalies in a Chinese population: a 10-year experience.

BACKGROUND: The 595-nm pulsed dye laser (PDL) has been used to treat vascular anomalies for about 30 years; however, there are insufficient data in Chinese patients concerning therapeutic efficacy, optimized parameters, and procedure techniques. OBJECTIVE: To study the efficacy and relevant factors in PDL therapy for vascular anomalies in Chinese patients. METHOD: We enrolled 431 patients with 8 different vascular anomalies and no previous treatment in this retrospective study. A detailed classification of vascular anomalies and various parameters and techniques of PDL were studied. The clinical outcomes were analysed using the Investigator Global Assessment. RESULTS: Improvements were significantly correlated with infantile haemangioma (IH) subtypes (p < 0.05). A significant correlation between efficacy and lesion colour, anatomical sites, and hypertrophic-type port-wine stain (PWS) was found (p < 0.05). There was no significant correlation between efficacy and age or sex (p > 0.05). CONCLUSION: PDL is an effective and safe therapeutic modality for managing vascular anomalies in Chinese patients. We determined that differentiating and identifying IH subtypes prior to treatment could be a useful parameter for predicting therapeutic results.  Lesion colour, sites, and hypertrophic changes in PWS are relevant therapeutic factors. PDL parameters and techniques differ according to the various vascular anomalies to achieve optimal results.

Identification of the ionotropic GABA receptor-like subunits from the striped stem borer, Chilo suppressalis Walker (Lepidoptera: Pyralidae).

Ionotropic γ-aminobutyric acid (GABA) receptors (GABARs) mediate rapid inhibitory neurotransmission in both vertebrates and invertebrates, and are important molecular targets of insecticides. However, components of insect GABARs remain elusive. In addition to CsRDL1 and CsRDL2, the complementary DNAs (cDNAs) of another two GABA receptor-like subunits, CsLCCH3 and Cs8916, were identified from the rice striped stem borer, Chilo suppressalis Walker in the present study. Both CsLCCH3 and Cs8916 subunits shared common structural features, such as a highly-conserved Cys-loop structure, six distinct regions involved in ligand binding (loops A-F), and four transmembrane domains (TM 1-4). Transcript analysis demonstrated that the relative mRNA expression levels of both CsLCCH3 and Cs8916 subunits were the highest in the ventral nerve cord. Regarding developmental stage, transcript levels of both subunits were highest in eggs. Injections of double-stranded RNAs (dsRNAs), including dsRDL1, dsRDL2, dsLCCH3, or ds8916, significantly reduced mRNA abundance after 24 and 48 h. However, no observable effects on the development of C. suppressalis were observed. Injection of dsRDL1 or dsRDL2 did significantly reduce the mortality of C. suppressalis treated with fluralaner. Our results indicated that CsRDLs mediated the susceptibility of C. suppressalis to fluralaner, whereas CsLCCH3 and CsL8916 did not. The current investigation enhances our knowledge of Lepidopteran GABARs and offers a molecular basis for the development of novel insecticides to control C. suppressalis.

High-Fidelity Single-Qubit Gates on Neutral Atoms in a Two-Dimensional Magic-Intensity Optical Dipole Trap Array.

As a conventional approach, optical dipole trap (ODT) arrays with linear polarization have been widely used to assemble neutral-atom qubits for building a quantum computer. However, due to the inherent scalar differential light shifts (DLS) of qubit states induced by trapping fields, the microwave-driven gates acting on single qubits suffer from errors on the order of 10^{-3}. Here, we construct a DLS compensated ODT array based upon a recently developed magic-intensity trapping technique. In such a magic-intensity optical dipole trap (MI-ODT) array, the detrimental effects of DLS are efficiently mitigated so that the performance of global microwave-driven Clifford gates is significantly improved. Experimentally, we achieve an average error of (4.7±1.1)×10^{-5} per global gate, which is characterized by randomized benchmarking in a 4×4 MI-ODT array. Moreover, we experimentally study the correlation between the coherence time and gate errors in a single MI-ODT with an optimum error per gate of (3.0±0.7)×10^{-5}. Our demonstration shows that MI-ODT array is a versatile platform for building scalable quantum computers with neutral atoms.

CRootBox: a structural-functional modelling framework for root systems.

Background and Aims: Root architecture development determines the sites in soil where roots provide input of carbon and take up water and solutes. However, root architecture is difficult to determine experimentally when grown in opaque soil. Thus, root architecture models have been widely used and been further developed into functional-structural models that simulate the fate of water and solutes in the soil-root system. The root architecture model CRootBox presented here is a flexible framework to model root architecture and its interactions with static and dynamic soil environments. Methods: CRootBox is a C++-based root architecture model with Python binding, so that CRootBox can be included via a shared library into any Python code. Output formats include VTP, DGF, RSML and a plain text file containing coordinates of root nodes. Furthermore, a database of published root architecture parameters was created. The capabilities of CRootBox for the unconfined growth of single root systems, as well as the different parameter sets, are highlighted in a freely available web application. Key results: The capabilities of CRootBox are demonstrated through five different cases: (1) free growth of individual root systems; (2) growth of root systems in containers as a way to mimic experimental setups; (3) field-scale simulation; (4) root growth as affected by heterogeneous, static soil conditions; and (5) coupling CRootBox with code from the book Soil physics with Python to dynamically compute water flow in soil, root water uptake and water flow inside roots. Conclusions: CRootBox is a fast and flexible functional-structural root model that is based on state-of-the-art computational science methods. Its aim is to facilitate modelling of root responses to environmental conditions as well as the impact of roots on soil. In the future, this approach will be extended to the above-ground part of the plant.

Observers' response to facial disfigurement from head and neck cancer.

OBJECTIVE: Our long-term goal is to develop a normative feedback intervention to support head and neck cancer patients in forming realistic expectations about how other people in non-social group settings will respond to their appearance. This study aimed to evaluate the relationship between observer ratings of facial disfigurement and observer ratings of emotional response when viewing photographs of faces of head and neck cancer patients. METHODS: Seventy-five (75) observers rated their emotional response to each of 144 facial photographs of head and neck cancer patients using the Self-Assessment-Manikin and rated severity of facial disfigurement on a 9-point scale. Body image investment of the observers was measured using the Appearance Schemas Inventory-Revised. A standardized multiple regression model was used to assess the relationship between observer ratings of facial disfigurement and observer ratings of emotional response, taking into consideration the age and sex of the patient depicted in the stimulus photograph, as well as the age, sex, and body image investment of the observer. RESULTS: Observers who had a strong emotional response to a patient's facial photograph tended to rate the patient's facial disfigurement as more severe (standardized regression coefficient ß = 0.328, P < 0.001). Sex and age of the observer had more influence on the rating of facial disfigurement than did the patient's demographic characteristics. Observers more invested in their own body image tended to rate the facial disfigurement as more severe. CONCLUSIONS: This study lays the groundwork for a normative database of emotional response to facial disfigurement.

Body image dissatisfaction in patients undergoing breast reconstruction: Examining the roles of breast symmetry and appearance investment.

OBJECTIVE: Reconstruction as part of treatment for breast cancer is aimed at mitigating body image concerns after mastectomy. Although algorithms have been developed to objectively assess breast reconstruction outcomes, associations between objectively quantified breast aesthetic appearance and patient-reported body image outcomes have not been examined. Further, the role of appearance investment in explaining a patient's body image is not well understood. We investigated the extent to which objectively quantified breast symmetry and patient-reported appearance investment were associated with body image dissatisfaction in patients undergoing cancer-related breast reconstruction. METHODS: Breast cancer patients in different stages of reconstruction (n = 190) completed self-report measures of appearance investment and body image dissatisfaction. Vertical extent and horizontal extent symmetry values, which are indicators of breast symmetry, were calculated from clinical photographs. Associations among breast symmetry, appearance investment, body image dissatisfaction, and patient clinical factors were examined. Multi-variable regression was used to evaluate the extent to which symmetry and appearance investment were associated with body image dissatisfaction. RESULTS: Vertical extent symmetry, but not horizontal extent symmetry, was associated with body image dissatisfaction. Decreased vertical extent symmetry (ß = -.19, P < .05) and increased appearance investment (ß = .45, P < .001) were significantly associated with greater body image dissatisfaction while controlling for clinical factors. CONCLUSIONS: Breast symmetry and patient appearance investment both significantly contribute to an understanding of patient-reported body image satisfaction during breast reconstruction treatment.