Ultrasonic-Assisted Extraction of Phenolic Compounds from Lonicera similis Flowers at Three Harvest Periods: Comparison of Composition, Characterization, and Antioxidant Activity.

Lonicera similis Hemsl. (L. similis) is a promising industrial crop with flowers rich in phenolic compounds. In this study, an ultrasound-assisted extraction (UAE) was designed to extract phenolic compounds from L. similis flowers (LSFs). A contrastive analysis on the phenolic compounds' yield and characterization and the antioxidant activity of the extracts at three harvest stages (PGS I, PGS II, and PGS III) are reported. The results indicate that the optimal conditions are a sonication intensity of 205.9 W, ethanol concentration of 46.4%, SLR of 1 g: 31.7 mL, and sonication time of 20.1 min. Under these optimized conditions, the TPC values at PGS I, PGS II, and PGS III were 117.22 ± 0.55, 112.73 ± 1.68, and 107.33 ± 1.39 mg GAE/g, respectively, whereas the extract of PGS I had the highest TFC (68.48 ± 2.01 mg RE/g). The HPLC analysis showed that chlorogenic acid, rutin, quercetin, isoquercitrin, and ferulic acid are the main components in the phenolic compounds from LSFs, and their contents are closely corrected with the harvest periods. LSF extracts exhibited a better antioxidant activity, and the activity at PGS I was significantly higher than those at PGS II and PGS III. The correlation analysis showed that kaempferol and ferulic acid, among the eight phenolic compounds, have a significant positive correlation with the antioxidant activity, while the remaining compounds have a negative correlation. Minor differences in extracts at the three harvest stages were found through SEM and FTIR. These findings may provide useful references for the optimal extraction method of phenolic compounds from LSFs at three different harvest periods, which will help to achieve a higher phytochemical yield at the optimal harvest stage (PGS I).

Extraction Optimization of Phenolic Compounds from Triadica sebifera Leaves: Identification, Characterization and Antioxidant Activity.

Triadica sebifera (T. sebifera) has attracted much attention because of the high oil content in its seeds, but there are few systematic studies on the phenolic compounds of T. sebifera leaves (TSP). In this study, the extraction process of TSP was optimized by response surface methodology. The phenolic components of these extracts were analyzed by high-performance liquid chromatography (HPLC). Moreover, the effects of hot air drying (HD), vacuum drying (VD) and freeze drying (FD) on the antioxidant activity and characterization of T. sebifera leaf extract (TSLE) were evaluated. Under the conditions of ethanol concentration 39.8%, liquid-solid ratio (LSR) 52.1, extraction time 20.2 min and extraction temperature 50.6 °C, the maximum TSP yield was 111.46 mg GAE/g dw. The quantitative analysis and correlation analysis of eight compounds in TSP showed that the type and content of phenolic compounds had significant correlations with antioxidant activity, indicating that tannic acid, isoquercitrin and ellagic acid were the main components of antioxidant activities. In addition, through DPPH and ABTS determination, VD-TSLE and FD-TSLE showed strong scavenging ability, with IC50 values of 138.2 µg/mL and 135.5 µg/mL and 73.5 µg/mL and 74.3 µg/mL, respectively. Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) infrared spectroscopy revealed small differences in the extracts of the three drying methods. This study lays a foundation for the effective extraction process and drying methods of phenolic antioxidants from T. sebifera leaves, and is of great significance for the utilization of T. sebifera leaves.

CRISPR/Cas9-mediated gene knockout of Sj16 in Schistosoma japonicum eggs upregulates the host-to-egg immune response.

Schistosomiasis is an important, neglected tropical disease. Schistosoma japonicum can evade host attacks by regulating the host's immunity, causing continuous infection. However, interactions between the host's immune system and S. japonicum are unclear. Our previous research found that the Sj16 protein isolated from S. japonicum has an anti-inflammatory effect in the host. However, the role of Sj16 in the regulation of host immunity in S. japonicum infection is not clear. Here, we applied the CRISPR/Cas9 technique to knockout Sj16 in S. japonicum eggs and investigated the effect of Sj16 in regulating host immunity. We found egg viability decreased after Sj16 knockout. In addition, we found granulomatous inflammation increased, the T-cell immune response enhanced and the immune microenvironment changed in mice model injected with Sj16-knockout eggs by tail vein. These findings suggested that S. japonicum could regulate host immunity through Sj16 to evade the host immune attack and cause continuous infection. In addition, we confirmed the application of CRISPR/Cas9-mediated gene reprogramming for functional genomics in S. japonicum.

Feasibility and self-perceived effectiveness of an online training program on dysphagia in residential aged care homes in mainland China.

BACKGROUND: With a rapidly aging population in mainland China, dysphagia has become one of the common geriatric disorders which creates a huge demand on speech and language therapists (SLTs). The major challenge is the shortage of SLTs in China. In addition, frontline practitioners in mainland China may not be well equipped with the knowledge and practical skills in dysphagia management due to lack of systematic training and the work nature. AIMS: This study evaluates the self-perceived effectiveness and feasibility of an online training program that aims to enhance the self-assessed knowledge and skills of SLTs providing dysphagia care in residential aged care homes. METHODS AND PROCEDURES: Sixteen SLTs working in a residential aged care homes in mainland China attended a three-hour pilot online training program which consists of didactic lecture and practical skills activity components. A total of 10 participants completed an online questionnaire one month after the training to evaluate the feasibility and effectiveness of this online training program. OUTCOMES AND RESULTS: The preliminary results demonstrated participants' self-perception of high training effectiveness in theoretical knowledge and practical skills. A majority of the participants perceived that the training enhanced their theoretical knowledge and all of them perceived that they acquired practical skills. All respondents were satisfied with the online training approach. They also highlighted the advantage and challenges of the online training approach. CONCLUSIONS AND IMPLICATIONS: Online training is an effective and feasible approach for theoretical knowledge and practical skills transfer in SLT training and could ultimately benefit the delivery of services for individuals with dysphagia in mainland China. WHAT THIS PAPER ADDS: What is already known on the subject Previous studies have shown that online training approach is as effective as face-to-face training in increasing professional knowledge. Online training programs may be more cost efficient and time efficient when compared with face-to-face training. What this study adds The present study provided preliminary evidence to support the feasibility and effectiveness of using online training on dysphagia for speech and language therapists working in residential aged care homes in mainland China. What are the clinical implications of this work? From the participants' perception, online training approach is effective and feasible in delivering theoretical knowledge and practical skills. It may be a better training approach for mainland China considering the lack of expertise and accessibility to training.

Applying computer-based language test to young children.

INTRODUCTION: This study aims at exploring the feasibility of applying a computer-based language test to young children aged 2-4 years. METHODS: Thirty-two Cantonese-speaking children, aged 2-4 years, were recruited from local kindergartens. All participants underwent assessment using both the computer-based and paper-pencil versions of the Macau Cantonese Language Screening Scale for Preschool Children, following a crossover study design. A short break of 15-30 minutes was provided between the two assessments. The data were analysed at three levels: the overall test, subcategory, and individual item levels. At the overall test and subcategory levels, data were analysed using the paired samples t-test and ICC. At the item level, the percentage of agreement and Cohen's kappa value were selected to assess the agreement of the two test formats. RESULTS: Excellent agreement was found for the overall test level, and good agreement was observed for four of the five subcategories. At the individual item level, 28 of the 35 items showed more than 80% agreement, and 16 items showed substantial to almost perfect agreement. CONCLUSION: These results suggest that the two test formats give similar total scores and subcategory scores for children aged 2-4. For children older than 2 years, 6 months, the agreement for matching items is as high as 83.68% (1318/1575). The computer-based test is thus highly recommended for this group of children. For children younger than 2 years, 6 months, a modified computer-based test is suggested to accommodate their needs.

Randomized Controlled Trial of Awake Transnasal Laser-Assisted Surgery for Benign Laryngeal Lesions.

OBJECTIVE: To compare functional and cost-effectiveness of awake transnasal laser assisted-surgery versus microlaryngeal surgery for benign laryngeal lesions. METHODS: This was a prospective non-inferiority randomized controlled trial conducted from May 2021 to December 2022 at two tertiary referral hospitals in Hong Kong. Patients were block-randomized to receive either awake transnasal laser-assisted surgery or microlaryngeal surgery, with post-operative follow-up in a multidisciplinary voice clinic for 1-year. Primary outcome was Voice Handicap Index (VHI-30). Secondary outcomes included operation time, complications, length of stay, peri-operative discomfort, recurrence, and medical costs. RESULTS: Sixty-one patients were randomized to either awake transnasal laser-assisted surgery (n = 30) and microlaryngeal surgery (n = 31). Both groups had comparable demographics and laryngeal pathologies. Both groups showed significant improvement of VHI-30 score over time and had comparable post-operative VHI-30. Awake transnasal laser-assisted surgery group had a significantly shorter length of stay (0.5 vs. 1 day) and less throat discomfort (2 vs. 4) compared to microlaryngeal surgery group. Intraoperative complications were more common in microlaryngeal surgery group (14.3% vs. 0%). Otherwise, both groups had similar operative time and recurrence rate. Cost-analysis showed a significantly lower hospital cost for awake transnasal laser-assisted surgery (USD 3090) compared to microlaryngeal surgery group (USD 5120). CONCLUSION: Awake transnasal laser-assisted surgery was safe, functionally non-inferior, as measured by VHI-30, to microlaryngeal surgery in managing benign laryngeal lesions, while superior to microlaryngeal surgery in peri-operative discomfort and medical costs. LEVEL OF EVIDENCE: 2 Laryngoscope, 134:3732-3740, 2024.

The transplant rejection response involves neutrophil and macrophage adhesion-mediated trogocytosis and is regulated by NFATc3.

The anti-foreign tissue (transplant rejection) response, mediated by the immune system, has been the biggest obstacle to successful organ transplantation. There are still many enigmas regarding this process and some aspects of the underlying mechanisms driving the immune response against foreign tissues remain poorly understood. Here, we found that a large number of neutrophils and macrophages were attached to the graft during skin transplantation. Furthermore, both types of cells could autonomously adhere to and damage neonatal rat cardiomyocyte mass (NRCM) in vitro. We have demonstrated that Complement C3 and the receptor CR3 participated in neutrophils/macrophages-mediated adhesion and damage this foreign tissue (NRCM or skin grafts). We have provided direct evidence that the damage to these tissues occurs by a process referred to as trogocytosis, a damage mode that has never previously been reported to directly destroy grafts. We further demonstrated that this process can be regulated by NFAT, in particular, NFATc3. This study not only enriches an understanding of host-donor interaction in transplant rejection, but also provides new avenues for exploring the development of novel immunosuppressive drugs which prevent rejection during transplant therapy.

Nitric oxide-induced endoplasmic reticulum stress of Schistosoma japonicum inhibits the worm development in rats.

Schistosomiasis, caused by Schistosoma spp., is a zoonotic parasitic disease affecting human health. Rattus norvegicus (rats) are a non-permissive host of Schistosoma, in which the worms cannot mature and cause typical egg granuloma. We previously demonstrated that inherent high levels of nitric oxide (NO), produced by inducible NO synthase (iNOS), is a key molecule in blocking the development of S. japonicum in rats. To further explore the mechanism of NO inhibiting S. japonicum development in rats, we performed S-nitrosocysteine proteomics of S. japonicum collected from infected rats and mice. The results suggested that S. japonicum in rats may have undergone endoplasmic reticulum (ER) stress. Interestingly, we found that the ER of S. japonicum in rats showed marked damage, while the ER of the worm in iNOS-/- rats and mice were relatively normal. Moreover, the expression of ER stress markers in S. japonicum from WT rats was significantly increased, compared with S. japonicum from iNOS-/- rats and mice. Using the NO donor sodium nitroprusside in vitro, we demonstrated that NO could induce ER stress in S. japonicum in a dose-dependent manner, and the NO-induced ER stress in S. japonicum could be inhibited by ER stress inhibitor 4-Phenyl butyric acid. We further verified that inhibiting ER stress of S. japonicum in rats promoted parasite development and survival. Furthermore, we demonstrated that NO-induced ER stress of S. japonicum was related to the efflux of Ca2+ from ER and the impairment of mitochondrial function. Collectively, these findings show that high levels of NO in rats could induce ER stress in S. japonicum by promoting the efflux of Ca2+ from ER and damaging the mitochondrial function, which block the worm development. Thus, this study further clarifies the mechanism of anti-schistosome in rats and provides potential strategies for drug development against schistosomiasis and other parasitosis.

Macrophage-mediated trogocytosis contributes to destroying human schistosomes in a non-susceptible rodent host, Microtus fortis.

Schistosoma parasites, causing schistosomiasis, exhibit typical host specificity in host preference. Many mammals, including humans, are susceptible to infection, while the widely distributed rodent, Microtus fortis, exhibits natural anti-schistosome characteristics. The mechanisms of host susceptibility remain poorly understood. Comparison of schistosome infection in M. fortis with the infection in laboratory mice (highly sensitive to infection) offers a good model system to investigate these mechanisms and to gain an insight into host specificity. In this study, we showed that large numbers of leukocytes attach to the surface of human schistosomes in M. fortis but not in mice. Single-cell RNA-sequencing analyses revealed that macrophages might be involved in the cell adhesion, and we further demonstrated that M. fortis macrophages could be mediated to attach and kill schistosomula with dependence on Complement component 3 (C3) and Complement receptor 3 (CR3). Importantly, we provided direct evidence that M. fortis macrophages could destroy schistosomula by trogocytosis, a previously undescribed mode for killing helminths. This process was regulated by Ca2+/NFAT signaling. These findings not only elucidate a novel anti-schistosome mechanism in M. fortis but also provide a better understanding of host parasite interactions, host specificity and the potential generation of novel strategies for schistosomiasis control.

Gnawing Between Cells and Cells in the Immune System: Friend or Foe? A Review of Trogocytosis.

Trogocytosis occurs when one cell contacts and quickly nibbles another cell and is characterized by contact between living cells and rapid transfer of membrane fragments with functional integrity. Many immune cells are involved in this process, such as T cells, B cells, NK cells, APCs. The transferred membrane molecules including MHC molecules, costimulatory molecules, receptors, antigens, etc. An increasing number of studies have shown that trogocytosis plays an important role in the immune system and the occurrence of relevant diseases. Thus, whether trogocytosis is a friend or foe of the immune system is puzzling, and the precise mechanism underlying it has not yet been fully elucidated. Here, we provide an integrated view of the acquired findings on the connections between trogocytosis and the immune system.

Recombinant protein Schistosoma japonicum-derived molecule attenuates dextran sulfate sodium-induced colitis by inhibiting miRNA-217-5p to alleviate apoptosis.

BACKGROUND: Inflammatory bowel disease (IBD) affects millions of people worldwide and has emerged as a growing problem in industrialized nations. The lack of therapeutic targets has limited the treatment of IBD. Studies found that parasitic nematode infections can ameliorate clinical and experimental colitis. Our previous study found that rSj16, a 16-kDa secreted protein of Schistosoma japonicum produced by Escherichia coli, has protective effects on dextran sulfate sodium (DSS)-induced colitis in mice. Apoptosis is an important factor in the pathogenesis of colitis. However, it is not clear whether the effect of rSj16 on colitis is related to apoptosis. AIM: To investigate whether the protective effects of rSj16 on colitis is related to apoptosis and its mechanism. METHODS: In-vivo , colitis was induced by DSS. The severity of colitis was assessed. WB was used to detect the changes of apoptosis-related genes in colon tissues. Q-PCR was used to detect the changes of miRNA-217-5p and HNF1B. In-vitro , WB was used to detect the changes of apoptosis-related genes in intestinal epithelial cells. TUNNEL staining and flow cytometry were used to detect cell apoptosis. RESULTS: rSj16 attenuates clinical activity in DSS-induced colitis mice. TUNNEL staining and WB results showed that apoptosis was increased in colon tissue after treatment with DSS, and the apoptosis of colon tissue was significantly reduced after treatment with rSj16. Compared with normal mice, the expression of miR-217-5p was increased in colon tissue of DSS-induced colitis mice. In addition, the miR-217-5p target gene hnf1b was decreased after administration of DSS. After treatment with rSj16, the expression of miR-217-5p was decreased and the expression of HNF1B was increased compared with the DSS-treated group. When Etoposide was used in combination with miR-217-5p mimic on MODE-K cells, the expression of cleaved-Caspase-3 and Bax was increased, and Bcl-2 was decreased compared with only Etoposide treatment, the expression of HNF1B was significantly reduced, suggesting that miR-217-5p acts as a pro-apoptotic in colon epithelial cells and down-regulates the target gene hnf1b. After rSj16 administration in MODE-K cells, miR-217-5p expression was significantly decreased, HNF1B expression was increased, and apoptosis was reduced. CONCLUSION: The protective effects of rSj16 on colitis is related to apoptosis and miRNA-217-5p may be a further target for therapeutic intervention against IBD.

Biohydrogen production in the suspended and attached microbial growth systems from waste pastry hydrolysate.

Waste pastry was hydrolyzed by glucoamylase and protease which were obtained from solid state fermentation of Aspergillus awamori and Aspergillus oryzae to produce waste pastry hydrolysate. Then, the effects of hydraulic retention times (HRTs) (4-12h) on hydrogen production rate (HPR) in the suspended microbial growth system (continuous stirred tank reactor, CSTR) and attached microbial growth system (continuous mixed immobilized sludge reactor, CMISR) from waste pastry hydrolysate were investigated. The maximum HPRs of CSTR (201.8mL/(h·L)) and CMISR (255.3mL/(h·L)) were obtained at HRT of 6h and 4h, respectively. The first-order reaction could be used to describe the enzymatic hydrolysis of waste pastry. The carbon content of the waste pastry remained 22.8% in the undigested waste pastry and consumed 77.2% for carbon dioxide and soluble microbial products. To our knowledge, this is the first study which reports biohydrogen production from waste pastry.

Effect of organic loading rate on dark fermentative hydrogen production in the continuous stirred tank reactor and continuous mixed immobilized sludge reactor from waste pastry hydrolysate.

Waste pastry (6%, w/v) was hydrolyzed by the produced glucoamylase and protease to obtain the glucose (19.8g/L) and free amino nitrogen (179mg/L) solution. Then, the effect of organic loading rate (OLR) (8-40kgCOD/(m3d)) on dark fermentative hydrogen production in the continuous stirred tank reactor (CSTR) and continuous mixed immobilized sludge reactor (CMISR) from waste pastry hydrolysate was investigated and compared. The maximum hydrogen production rate of CSTR (277.76mL/(hL)) and CMISR (320.2mL/(hL)) were achieved at OLR of 24kgCOD/(m3d) and 32kgCOD/(m3d), respectively. Carbon recovery ranged from 75.2-84.1% in the CSTR and CMISR with the balance assumed to be converted to biomass. One gram waste pastry could produce 0.33g (1.83mmol) glucose which could be further converted to 79.24mL (3.54mmol) hydrogen in the CMISR or 91.66mL (4.09mmol) hydrogen in the CSTR. This is the first study which reports dark fermentative hydrogen production from waste pastry.