Effect of electron beam irradiation on raw goat milk: microbiological, physicochemical and protein structural analysis.

BACKGROUND: Goat milk is considered a nutritionally superior resource, owing to its advantageous nutritional attributes. Nevertheless, it is susceptible to spoilage and the persistence of pathogens. Electron beam irradiation stands as a promising non-thermal processing technique capable of prolonging shelf life with minimal residue and a high degree of automation. RESULTS: The effects of electron beam irradiation (2, 3, 5, and 7 kGy) on microorganisms, physicochemical properties, and protein structure of goat milk compared with conventional pasteurized goat milk (PGM) was evaluated. It was found that a 2 kGy electron beam irradiation reduces the total microbial count of goat milk by 6-logs, and the irradiated goat milk protein secondary structure showed a significant decrease in ɑ-helix content. Low irradiation doses led to microaggregation and crosslinking. In contrast, high doses (≥ 5 kGy) slightly disrupted the aggregates and decreased the particle size, disrupting the microscopic surface structure of goat milk, verified by scanning electron microscopy and confocal laser scanning microscopy. CONCLUSION: The irradiation of goat milk with a 2 kGy electron beam may effectively inactivate harmful microorganisms in the milk and maintain/or improve the physicochemical quality and protein structure of goat milk compared to thermal pasteurization. © 2024 Society of Chemical Industry.

Clinicopathological differences and survival benefit in ER+/PR+/HER2+ vs ER+/PR-/HER2+ breast cancer subtypes.

INTRODUCTION: Breast cancer subtypes based on estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) expression have significant implications for prognosis. HER2-positive tumors historically demonstrated poorer survival, but anti-HER2 targeted therapy improved outcomes. However, hormone receptor (HR)-positive patients may experience reduced benefit due to HER2-HR signaling crosstalk. METHODS: Data from two databases, the Shanghai Jiao Tong University Breast Cancer Data Base (SJTUBCDB) and the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) database, were analyzed. Propensity score adjustments were used to balance patient characteristics between ER+/PR+/HER2+ and ER+/PR-/HER2+ subtypes. Kaplan-Meier survival curves estimated disease-free survival (DFS), breast cancer-specific survival (BCSS), overall survival (OS) for these subtypes in the SJTUBCDB, while subgroup analyses using multivariable models were performed based on menstruation, pN stage, HER2-targeted therapy, and endocrinotherapy. RESULTS: The ER+/PR+/HER2+ group showed significantly better DFS and BCSS than the ER+/PR-/HER2+ group, particularly in postmenopausal and pN0 stage patients. Survival outcomes were similar after anti-HER2 therapy or endocrine aromatase inhibitor (AI) therapy in both groups. However, among patients receiving selective estrogen receptor modulator (SERM) treatment, those in the ER+/PR-/HER2+ group had a significantly worse prognosis compared to ER+/PR+/HER2+ patients. CONCLUSIONS: HER2-positive breast cancers with different HR statuses exhibit distinct clinicopathological features and survival outcomes. Patients in the ER+/PR+/HER2+ group generally experience better survival, particularly in postmenopausal and pN0 stage patients. Treatment strategies should consider HR status and specific modalities for better personalized management.

Effect of electron beam irradiation on the structural characteristics and functional properties of goat's milk casein.

The effects of electron beam irradiation (EBI) at different doses (0, 2, 4, 6, 8, and 10 kGy) were investigated on the structural and functional properties of casein, including their interrelationship. A gradual reduction in the α-helix content of the secondary structure (as a stable structure) indicates that casein under EBI treatment mainly undergoes fragmentation and aggregation from a structural perspective. Furthermore, the hydrophobic group and tryptophan in the tertiary structure were exposed, which opened up the internal structure of the protein. In addition, a continuously increasing irradiation dose led to casein aggregation, as confirmed by electron microscopy. The structural changes affected its functional properties, such as solubility, emulsification, foaming, and rheological properties, all of which increased first and subsequently decreased. Finally, at irradiation doses of 4-6 kGy, casein was modified to exhibit optimal functional properties, which enhanced its food processing value and performance.

Electron beam irradiation induced aggregation, structural and functional changes of soybean 11S globulin.

This study investigated the effects of different irradiation doses of an electron beam (e-beam) (0, 2, 4, 6, 8, and 10 kGy) on the structure, emulsification, foaming, and rheological and gel properties of soybean 11S globulin. The irradiation treatment at 4 and 6 kGy significantly increased the solubility, surface hydrophobicity, disulfide bonding, and ζ-potential of 11S globulin, decreased the particle size of the protein solution, and effectively improved the emulsifying activity and foaming stability of the protein solution. Moreover, irradiation induced moderate cross-linking and aggregation of the proteins, thereby increasing the apparent viscosity and shear stress of the protein solution. In addition, the low-field NMR and microstructure analysis results revealed that protein gels formed a dense and homogeneous three-dimensional mesh structure after irradiation (6 kGy), along with increased content of bound water (T2b) and water not readily flowable (T21) and a decrease content of free water (T22). Overall, our results confirmed that e-beam irradiation could significantly improve the physicochemical properties of soybean 11S globulin. Our study thus provides a new technical means for the application of electron beam irradiation technology toward protein modification and broadens the high-value utilization of soybean 11S globulin in the food processing industry.

Chitosan/konjac glucomannan bilayer films: Physical, structural, and thermal properties.

To overcome the limitations of chitosan (CS) and konjac glucomannan (KGM), the bilayer films of CS and KGM were prepared by layer-by-layer (LBL) casting method, and the effects of different mass ratios (i.e., C5: K0, C4:K1, C3:K2, C1:K1, C2:K3, C1:K4, and C0:K5) on the microstructures and physicochemical properties of bilayer films were examined to evaluate their applicability in food packaging. The results revealed that the bilayer films had uniform microstructures. When compared with pure films, the bilayer films displayed lower swelling degrees and water vapor permeability. However, the tensile tests revealed a reduction in the mechanical properties of the bilayer films, which was nonetheless superior to that of the pure KGM film. In addition, the intermolecular interactions between the CS and KGM layers were observed through FTIR and XRD analyses. Finally, TGA and DSC analyses demonstrated a decrease in the thermal stability of the bilayer films. Our cumulative results verified that CS-KGM bilayer films may be a promising material for use in food packaging and further properties of the bilayer films can be supplemented in the future through layer-by-layer modification and the addition of active ingredients.

Survival outcomes of low-risk papillary thyroid carcinoma at different risk levels: a corollary for active surveillance.

Background: This study aims to compare the outcomes of active surveillance (AS) in low-risk papillary thyroid carcinoma (PTC) patients with different tumor sizes and lymph node metastasis status, in order to establish appropriate management strategies. By analyzing these results, this study provides valuable insights for the effective management of such patients, addressing the issues and challenges associated with AS in practical clinical practice. Methods: The study utilized the SEER database supported by the National Cancer Institute of the United States, extracting data of PTC diagnosed between 2000 and 2015. Statistical analyses were conducted using inverse probability weighting (IPTW) and propensity score matching (PSM), including Kaplan-Meier survival curves and Cox regression models, to evaluate the impact of different tumor sizes and lymph node metastasis status on thyroid cancer-specific survival (TCSS). Results: A total of 57,000 PTC patients were included, with most covariates having standardized mean differences below 10% after IPTW and PSM adjustments. The TCSS of PTC with a diameter smaller than 13mm is significantly better than that of tumors with a diameter larger than 13mm, regardless of the presence of lymph node metastasis. Among PTC cases with a diameter smaller than 13mm, the TCSS of patients is similar, regardless of the presence of lymph node metastasis. However, in PTC cases with a diameter larger than 13mm, the presence of lateral neck lymph node metastasis (N1b stage) significantly impacts the TCSS, although the absolute impact on TCSS rate is minimal. Conclusion: The treatment strategy of AS is safe for patients with T1a stage papillary thyroid microcarcinoma (PTMC). However, for patients with T1b stage, if the tumor diameter exceeds 13mm or there is lymph node metastasis in the lateral neck region, the TCSS will be significantly affected. Nevertheless, the absolute impact on survival is relatively small.

Enhancing the Electrical Conductivity and Strength of PET by Single-Wall Carbon Nanotube Film Coating.

Single-wall carbon nanotubes (SWCNTs) with excellent physicochemical properties are considered a promising candidate for the electrical and mechanical reinforcements of polymers. However, the poor dispersion of SWCNTs in plastics seriously limits their application and their achieved performance enhancement. Here, we coat a freestanding, highly conductive SWCNT film onto the surface of a polyethylene terephthalate (PET) film by a hot-pressing method. Due to the uniform SWCNT network structure and strong interfacial interaction, the SWCNT/PET hybrid film showed notably enhanced electrical and mechanical properties even though with a very low SWCNT weight fraction of 0.066%. The surface square resistance of the SWCNT/PET film decreased to 120-140 Ω/□ from 1016 Ω. In addition, Young's modulus and tensile strength of the SWCNT/PET film reached 4.6 GPa and 148 MPa, which are 31.3 and 24.4%, respectively, higher than the pure PET film. The SWCNT/PET film shows excellent mechanical durability and thermal stability, demonstrating its potential use as an antistatic material.

A multifunctional supramolecular assembly based on cucurbit[7]uril: White light material and Fe(CN)63- detection.

White light emitting materials have broad application prospects in fields such as displays, lighting devices, etc., but developing such materials faces considerable challenges. In this study, 1,3,5-tris[4-(pyridine-4-butyl)phenyl]benzene derivative (BTPY) was synthesized and a supramolecular assembly with AIE properties named BTPY@Q[7] was prepared with cucurbit[7]uril (Q[7]). Furthermore, by adding rhodamine 6G (R6G) to it, and controlling its ratio with R6G, a dual-emission white light system (0.33, 0.33) was synthesized and used for white light emitting materials as well as anti-counterfeiting fields. In addition, based on the BTPY@Q[7]-R6G system, a light harvesting system in aqueous phase was constructed, with an energy transfer efficiency (ΦET) of 26.19 % and an antenna effect (AE) of 10.21. Interestingly, the supramolecular self-assembly can also be used as a fluorescent probe, specifically recognize Fe(CN)63- ions in water, with a detection limit of 2.5 × 10-8 M.

Survival Benefit of Adjuvant Chemotherapy in Node-Positive Breast Cancer With a 21-Gene Recurrence Score of 14 to 25: A Real-World Study Based on the Inverse Probability of Treatment Weighting Method.

INTRODUCTION: The role of recurrence score in predicting the benefits of adjuvant chemotherapy for lymph-node-positive breast cancer remains uncertain. We studied chemotherapy usage in patients with 1 to 3 positive lymph nodes and a recurrence score (RS) of 25 or lower to assess changes in clinical practice based on the RxPONDER trial. METHODS: A retrospective study using the SEER database identified female patients diagnosed with ER-positive, HER2-negative breast cancer, 1 to 3 positive lymph nodes, and an RS of 25 or lower between 2010 and 2015. Patients were divided into nonchemotherapy and chemotherapy groups, with propensity score weighting to balance clinicopathologic factors. RESULTS: Among 7965 patients, 5774 (72.5%) were in the nonchemotherapy group, while 2191 (27.5%) were in the chemotherapy group. Median follow-up was 39 months. Breast cancer accounted for 67 deaths, while 128 deaths were due to other causes. The weighted 5-year overall survival (OS) rates were 95.7% for the nonchemotherapy group and 97.2% for the chemotherapy group. For high-risk patients, the weighted 5-year OS rates were 95.2% and 97.0% for the nonchemotherapy and chemotherapy groups, respectively, with a significant absolute difference of 1.8% (P = .014). Multivariate analysis showed a significant difference in weighted hazard ratios for OS between the nonchemotherapy and chemotherapy groups in high-risk patients (hazard ratio: 0.64; 95% CI: 0.48-0.86). However, there were no significant differences in weighted hazard ratios for lower-risk patients, and similar results were observed for breast cancer-specific survival (BCSS). CONCLUSION: Patients with ER-positive, HER2-negative breast cancer and 1 to 3 positive lymph nodes, assessed by a 21-gene RS of 0 to 25, exhibited heterogeneous prognosis. Adjuvant chemotherapy provided a significant survival benefit, especially for patients with RS of 14 to 25, particularly those with invasive ductal carcinoma (IDC) and 2 to 3 positive lymph nodes.

Evaluation of effects of ultrasound-assisted curing on the flavor of Chinese bacon.

The curing stage is of great importance in flavor formation during Chinese bacon processing. Ultrasound-assisted curing plays an essential role in the Lipid oxidation of meat products. In this study, GC-MS and electronic nose were used to analyze the influence of different power ultrasonic-assisted curing on the flavor formation of Chinese bacon. Through the analysis of phospholipid and lipase, the fundamental precursors of ultrasonic on the flavor of Chinese bacon were determined. It was found that there were differences in the flavor contour description of Chinese bacon between the ultrasonic treatment group, mainly due to the change in the W1W sensor. A total of 28 volatile compounds were detected by GC-MS, and the aldehyde content increased with ultrasonic power. PC and PE are the main flavor precursors in the curing process. This study provides a theoretical basis for improving the curing technology of Chinese bacon.

Magnetic resonance imaging in screening women at high risk of breast cancer: A meta-analysis.

BACKGROUND: Magnetic resonance imaging (MRI) is more accurate than mammography in screening for breast cancer. Exposure to ionizing radiation from repeated diagnostic X-rays may be a cause of breast cancer. METHODS: We conducted systematic searches on PubMed, Cochrane and Embase to identify studies on women who underwent mammography or MRI screening. A meta-analysis was performed to compare the detection rate of breast cancer by mammography, MRI or both. RESULTS: A total of 18 diagnostic publications were identified and included in the meta-analysis. Among the 1000 screened women, MRI alone increased the detection rate of breast cancer by 8 compared with mammography alone (RR 0.48, 95% CI 0.42-0.54), and MRI plus mammography increased the detection rate of breast cancer by 1 compared with MRI alone (RR 0.86, 95% CI 0.78-0.96). Subgroup analysis demonstrated that the diagnostic efficacy of MRI plus mammography in breast was obviously better than that of MRI alone or mammography alone. CONCLUSIONS: Screening with MRI alone might be the best choice for women at high risk of breast cancer.

Integrated proteomics and metabolomics analysis revealed the mechanisms underlying the effect of irradiation on the fat quality of Chinese bacon.

Irradiation increases the security and storage period of preserved Chinese bacon; nevertheless, the biological mechanisms underlying the changes in fat quality caused by irradiation are unknown. We investigated the influence of irradiation on Chinese bacon by proteomic and metabolomic. We identified 24 proteins that participated in metabolism and 40 common differential metabolites enriched in 16 signalling pathways. Correlation analysis revealed that irradiation altered 11 pathways shared between the proteome and metabolome, including two lipid metabolism pathways. Acetyl-CoA carboxylase, ACSL, octanoic acid, decanoic acid, palmitic acid, and oleic acid participated in fatty acid biosynthesis. Acyl-CoA thioesterase 1/2/4, enoyl-CoA reductase, acetyl-CoA acyltransferase 1, enoyl-CoA hydratase 2, palmitic acid, and oleic acid participated in unsaturated fatty acid biosynthesis. These findings lay the groundwork for multi-omics research on the effects of irradiation on Chinese bacon quality, assisting in assessing irradiated Chinese bacon quality, and developing effective strategies to standardise quality parameters.

Application of the Luminescent luxCDABE Gene for the Rapid Screening of Antibacterial Substances Targeting Pseudomonas aeruginosa.

Pseudomonas aeruginosa (P. aeruginosa) is a typical Gram-negative bacterium that can cause the spoilage of catered food products. Using a luminescent reporter gene (luxCDABE), this study sought to construct a cell-based biosensor (PAO1-CE) to rapidly screen antibacterial substances against P. aeruginosa. A total of six antibiotics belonging to five categories were used as the model test substances. The results of the bioluminescence detection method were verified using traditional antibacterial research assessments. The correlation coefficient of the regression equation fitting the data generated using this method was greater than 0.98, supporting the credibility of this approach. Additionally, the EC50 of each of the antibiotics assessed in this study was lower than the 1/2 MIC determined by conventional means. All six of the antibiotics caused varying degrees of damage to the cell membrane and cell wall of P. aeruginosa. Importantly, this novel method helped shorten the time necessary for active-compound detection and could be used for high-throughput detection, which would also help improve the detection efficiency. The application of this method towards the discovery of novel antibacterial compounds targeting P. aeruginosa holds substantial promise for greatly improving the efficiency of compound discovery.

Antimicrobial effect of sorbic acid-loaded chitosan/tripolyphosphate nanoparticles on Pseudomonas aeruginosa.

Sorbic acid-loaded chitosan/tripolyphosphate nanoparticles (SANs) have previously been shown to exert both antibacterial and antioxidant effects on Chinese sausage. In this study, the minimum inhibitory concentrations (MICs) of SANs against two Pseudomonas aeruginosa strains were determined. The blank control group (BC) served as the negative control, while the chitosan/tripolyphosphate nanoparticles (CTNs) group and free sorbic acid (SA) group served as the positive controls. Tests conducted under five different pH conditions (5/6/7/8/9) revealed that the SANs exhibited a good bacteriostatic effect against P. aeruginosa. Variations in the metabolism, cell membrane or cell wall integrity, and morphology of P. aeruginosa were measured to evaluate the effects of SANs on their intracellular and extracellular components. The MIC of SANs for the two P. aeruginosa strains was determined to be 150 µg/mL. SANs delayed the growth of P. aeruginosa and severely damaged both its inner and outer cell membranes. The heteromorphism of the bacteria as observed by field emission scanning electron microscopy (FESEM), verified the aforementioned results. The results showed SANs could effectively inhibit the growth of P. aeruginosa and exert antibacterial ability in a wider range of acid-base environments. This study broadens the application of SANs in food processing and provides experimental basis.

Progress in arthroscopic surgery for injury of superior labrum anterior posterior of shoulder joint / 中国骨伤

Superior labrum anterior posterior (SLAP) injury is a major challenge for orthopedic surgeons, due to the poor healing ability of the injured labrum. Although arthroscopic surgery is the gold standard for the treatment of SLAP injury, there are still disputes about the adaptation of different surgical techniques, the choice of anchors during operation, knotted or knotless anchors, and fixation methods. The authors believe that arthroscopic repair of SLAP lesions is effective for young patients with intact glenoid labrum(<35 years old) or with extensive activity, where single and knotless anchor is preferred. For the older patients(≥35 years old) with degeneration and wear of glenoid labrum, biceps tenodesis is more preferable, and interference screw fixation technique is recommended. As for patients with failed SLAP repair, biceps tenodesis can achieve a high success rate as a revision surgery. By review of the relevant literature in recent years, this paper summarizes the adaptation of different surgical methods of arthroscopic treatment of SLAP injury, intraoperative anchoring techniques, fixation methods and other improved surgical techniques.

Evaluation of the Influence of Flavor Characteristics of Cooked Bacon with Different Sterilization Methods by GC-IMS Combined with HS-SPME-GC-MS and Electronic Nose.

This study investigated the impact of high pressure and temperature (HTHP) and electron-beam irradiations (3, 5, 7, and 9 kGy) using differences in two sterilization methods on the volatile compounds and sensory characteristics of cooked bacon. It showed that 7 and 9 kGy of irradiation caused a significant reduction in species of volatile compounds and sensory features, but the concentration of total ketones, alcohols, aldehydes, acids and aromatic hydrocarbons significantly increased at 9 kGy. Samples treated with a dose of less than 5 kGy did not change volatile compounds and sensory properties. High-temperature-high-pressure conditions could greatly impact the concentrations of volatile compound species and sensory traits. The electronic nose effectively detected the flavor difference in different sterilization methods. Fingerprinting showed that HTHP and 9-kGy-treated groups were significantly different from other treatments. This study inferred that 5 kGy might be optimal for maintaining the original flavor and sensory properties of cooked bacon.

Progress on arthroscopic surgery for massive rotator cuff tears / 中国骨伤

The surgical treatment of massive rotator cuff tears is a clinical challenge for orthopaedic surgeons. Moreover, tendon retraction, adhesions and fatty infiltration after rotator cuff tear will further increase the difficulty of surgical repair. Therefore, it has become a hotspot and difficulty to repair massive rotator cuff tears with a better way in current research. In recent years, with the continuous development of arthroscopic techniques, shoulder arthroscopic surgery has become the gold standard for the treatment of massive rotator cuff tears, but the adaptations, effects and combined application of different surgical methods are still controversial. The author believes that arthroscopic debridement of shoulder joint and acromioplasty or tuberoplasty could relieve shoulder pain in the short-term for elderly patients with lower functional requirements;long biceps tenotomy or tenodesis is effective for patients with biceps long head tendon injury; complete repair is still the first line treatment for massive rotator cuff tears, but partial repair is possible for massive rotator cuff tears that could not be completely repaired;patch augmentation technology could bring good results for young patients with high functional requirements;for patients with limited internal and external rotation of the shoulder joint and high functional requirements, tendon transfers surgery is recommended;superior capsular reconstruction is more advantageous for young patients with no obvious glenohumeral arthritis, better deltoid muscle strength and higher functional requirements. In addition, subacromial spacer implantation has become a current research hotspot due to its advantages of small trauma, low cost and relative safety, and its long-term effect still needs to be further confirmed.

The application of nano-biosensors in the detection of 2019-nCoV / 中华检验医学杂志

The pandemic of 2019 novel coronavirus (2019-nCoV) infection since 2020 caused Coronavirus Disease 2019 (COVID-19) leads the serious threaten to global public health. It is urgent to diagnose COVID-19, guide epidemiological measures, control the infection rates, research/develop the antiviral treatment and promote the vaccine research. The application of nano-material based biosensors (the nano-biosensors) has achieved the high-performance detection of a variety of biomarkers due to their small device size, label free detection, high sensitivity, good specificity, short detection time, and has been considered as great potential to become a point-of-care testing tool for detecting 2019-nCoV. Therefore, by summarizing the working principle and classification of nano-biosensors, and focusing on the research progress of nano-biosensors in the detection of 2019-nCoV reported in the recent years, our review provides the challenges and future development prospects of the nano-biosensor in clinical laboratory.

Characterization of the Volatile Compounds of Zhenba Bacon at Different Process Stages Using GC-MS and GC-IMS.

Zhenba bacon (ZB), a type of Chinese traditional bacon with a long history, has attracted considerable attention in the Southwest of China for its unique flavor. To elucidate the changing course of aroma components during the process of ZB, four stages of process stages were assessed by GC-MS and GC-IMS coupled with multivariate data analysis. A total of 44 volatile compounds were identified by GC-IMS, including 5 esters, 8 alcohols, 12 aldehydes, 3 ketones, 1 furan and 2 sulfides; 40 volatile compounds were identified by GC-MS, 4 ketones, 7 phenols, 8 alcohols, 6 esters, 6 aldehydes, and 6 other compounds were detected. During the curing period, the amount and content of esters in Zhenba bacon gradually increased. Phenols appear in large quantities during the smoking period. The VOCs (volatile organic compounds) in the gallery plots were the most diverse in YZ samples, which are mainly esters. POV (peroxide value) and TBARS (thiobarbituric acid reactive substance) showed that lipid oxidation played an important role in the formation of volatile flavor components of bacon. This study provides valuable analytical data to explain the flavor formation of Zhenba bacon.

Therapeutic Hypothermia Mitigates the Sepsis-Increased Permeability in EA. hy926 Cells by Preserving Rap1 Expression.

To determine the effect and potential mechanisms of therapeutic hypothermia (TH) on the permeability of septic cells. Human EA. hy926 cells were transfected with, or without, control or ras-proximate-1 (Rap1)-specific siRNA and treated with 2 µg/mL of lipopolysaccharide (LPS). The cells were cultured in normal temperature (NT) or a temporary TH for 10 hours. The cellular permeability of each group of cells was determined by transwell permeability assay. The relative levels of Rap1, RhoA (a small GTP enzyme of the Rho family), VE-cadherin expression, and myosin light chain (MLC) phosphorylation were quantified by Western blot and immunofluorescent assays. Compared with the control group, LPS stimulation increased cellular permeability in EA. hy926 cells under an NT condition, but significantly mitigated by TH. The effect of TH decreased after Rap1 silencing. Furthermore, LPS upregulated RhoA expression and MLC phosphorylation, but reduced Rap1 and VE-cadherin expression, which were also enhanced by Rap1 silencing, but significantly mitigated by TH. Immunofluorescent analyses indicated that LPS significantly increased phosphorylated MLC, but decreased VE-cadherin expression, which were further deteriorated by Rap1 silencing, but significantly mitigated by TH in EA. hy926 cells. TH significantly mitigated the sepsis-increased permeability of EA. hy926 cells by enhancing the Rap1 expression to attenuate the RhoA/MLC signaling.