Engineering a non-model yeast Rhodotorula mucilaginosa for terpenoids synthesis.

Terpenoids have tremendous biological activities and are widely employed in food, healthcare and pharmaceutical industries. Using synthetic biology to product terpenoids from microbial cell factories presents a promising alternative route compared to conventional methods such as chemical synthesis or phytoextraction. The red yeast Rhodotorula mucilaginosa has been widely studied due to its natural production capacity of carotenoid and lipids, indicating a strong endogenous isoprene pathway with readily available metabolic intermediates. This study constructed several engineered strains of R. mucilaginosa with the aim of producing different terpenoids. Monoterpene α-terpineol was produced by expressing the α-terpineol synthase from Vitis vinifera. The titer of α-terpineol was further enhanced to 0.39 mg/L by overexpressing the endogenous rate-limiting gene of the MVA pathway. Overexpression of α-farnesene synthase from Malus domestica, in combination with MVA pathway rate-limiting gene resulted in significant increase in α-farnesene production, reaching a titer of 822 mg/L. The carotenoid degradation product ß-ionone was produced at a titer of 0.87 mg/L by expressing the ß-ionone synthase from Petunia hybrida. This study demonstrates the potential of R. mucilaginosa as a platform host for the direct biosynthesis of various terpenoids and provides insights for further development of such platforms.

Ensemble learning with speaker embeddings in multiple speech task stimuli for depression detection.

Introduction: As a biomarker of depression, speech signal has attracted the interest of many researchers due to its characteristics of easy collection and non-invasive. However, subjects' speech variation under different scenes and emotional stimuli, the insufficient amount of depression speech data for deep learning, and the variable length of speech frame-level features have an impact on the recognition performance. Methods: The above problems, this study proposes a multi-task ensemble learning method based on speaker embeddings for depression classification. First, we extract the Mel Frequency Cepstral Coefficients (MFCC), the Perceptual Linear Predictive Coefficients (PLP), and the Filter Bank (FBANK) from the out-domain dataset (CN-Celeb) and train the Resnet x-vector extractor, Time delay neural network (TDNN) x-vector extractor, and i-vector extractor. Then, we extract the corresponding speaker embeddings of fixed length from the depression speech database of the Gansu Provincial Key Laboratory of Wearable Computing. Support Vector Machine (SVM) and Random Forest (RF) are used to obtain the classification results of speaker embeddings in nine speech tasks. To make full use of the information of speech tasks with different scenes and emotions, we aggregate the classification results of nine tasks into new features and then obtain the final classification results by using Multilayer Perceptron (MLP). In order to take advantage of the complementary effects of different features, Resnet x-vectors based on different acoustic features are fused in the ensemble learning method. Results: Experimental results demonstrate that (1) MFCC-based Resnet x-vectors perform best among the nine speaker embeddings for depression detection; (2) interview speech is better than picture descriptions speech, and neutral stimulus is the best among the three emotional valences in the depression recognition task; (3) our multi-task ensemble learning method with MFCC-based Resnet x-vectors can effectively identify depressed patients; (4) in all cases, the combination of MFCC-based Resnet x-vectors and PLP-based Resnet x-vectors in our ensemble learning method achieves the best results, outperforming other literature studies using the depression speech database. Discussion: Our multi-task ensemble learning method with MFCC-based Resnet x-vectors can fuse the depression related information of different stimuli effectively, which provides a new approach for depression detection. The limitation of this method is that speaker embeddings extractors were pre-trained on the out-domain dataset. We will consider using the augmented in-domain dataset for pre-training to improve the depression recognition performance further.

Dual-Stream Multiple Instance Learning for Depression Detection With Facial Expression Videos.

Depression is a common mental illness which has brought great harm to the individuals. With recent evidence that many objective physiological signals are associated with depression, automated detection of depression is urgent and important for the growing concern of mental illness. We investigate the problem of classifying depression by facial expressions, which may aid in online diagnosis and rehabilitation engineering of depression. In this work, We propose a weakly supervised learning approach employing multiple instance learning (MIL) on 150 videos data from 75 depressed and 75 healthy subjects. In addition, we present a novel MIL dual-stream aggregator that considers both the instance-level and the bag-level in order to emphasize the information with symptoms. Specifically, our method named ADDMIL uses max-pooling at the instance level to capture symptom information and further integrates the contribution of each instance at the bag level using attention weights. Our method achieves 74.7% accuracy and 74.5% recall on the collected dataset, which not only improves 10.1% accuracy and 9.8% recall over the baseline but also exceeds the best accuracy result of MIL-based method by 2.1%. Our work achieves results that are comparable to the state-of-the-art methods and demonstrates that multiple instance learning has great potential for depression classification. We present for the first time a weakly supervised learning approach in the detection of depression through raw facial expressions, which may provide a new framework for other psychiatric disorders detection methods.

Modularly engineering Rhodotorula toruloides for α-terpineol production.

α-Terpineol is a monoterpenoid alcohol that has been widely used in the flavor, fragrance, and pharmaceutical industries because of its sensory and biological properties. However, few studies have focused on the microbial production of α-terpineol. The oleaginous yeast Rhodotorula toruloides is endowed with a natural mevalonate pathway and is a promising host in synthetic biology and biorefinery. The primary objective of this work was to engineer R. toruloides for the direct biosynthesis of α-terpineol. The improvement in monoterpenoid production was achieved through the implementation of modular engineering strategies, which included the enhancement of precursor supply, blocking of downstream pathways, and disruption of competing pathways. The results of these three methods showed varying degrees of favorable outcomes in enhancing α-terpineol production. The engineered strain 5L6HE5, with competitive pathway disruption and increased substrate supply, reached the highest product titer of 1.5 mg/L, indicating that reducing lipid accumulation is an efficient method in R. toruloides engineering for terpenoid synthesis. This study reveals the potential of R. toruloides as a host platform for the synthesis of α-terpineol as well as other monoterpenoid compounds.

Multi-Center in-Depth Screening of Neonatal Deafness Genes: Zhejiang, China.

PURPOSE: The conventional genetic screening for deafness involves 9-20 variants from four genes. This study expands screening to analyze the mutation types and frequency of hereditary deafness genes in Zhejiang, China, and explore the significance of in-depth deafness genetic screening in newborns. METHODS: This was a multi-centre study conducted in 5,120 newborns from 12 major hospitals in the East-West (including mountains and islands) of Zhejiang Province. Concurrent hearing and genetic screening was performed. For genetic testing, 159 variants of 22 genes were screened, including CDH23, COL11A1, DFNA5, DFNB59, DSPP, GJB2, GJB3, KCNJ10, MT-RNR1, MT-TL1, MT-TS1, MYO15A, MYO7A, OTOF, PCDH15, SLC26A4, SOX10, TCOF1, TMC1, USH1G, WFS1, and WHRN using next-generation sequencing. Newborns who failed to have genetic mutations or hearing screening were diagnosed audiologically at the age of 6 months. RESULTS: A total of 4,893 newborns (95.57%) have passed the initial hearing screening, and 7 (0.14%) have failed in repeated screening. Of these, 446 (8.71%) newborns carried at least one genetic deafness-associated variant. High-risk pathogenic variants were found in 11 newborns (0.21%) (nine homozygotes and two compound heterozygotes), and eight of these infants have passed the hearing screening. The frequency of mutations in GJB2, GJB3, SLC26A4, 12SrRNA, and TMC1 was 5.43%, 0.59%, 1.91%, 0.98%, and 0.02%, respectively. The positive rate of in-depth screening was significantly increased when compared with 20 variants in four genes of traditional testing, wherein GJB2 was increased by 97.2%, SLC26A4 by 21% and MT-RNR1 by 150%. The most common mutation variants were GJB2c.235delC and SLC26A4c.919-2A > G, followed by GJB2c.299_300delAT. Homoplasmic mutation in MT-RNR1 was the most common, including m.1555A > G, m.961T > C, m.1095T > C. All these infants have passed routine hearing screening. The positive rate of MT-RNR1 mutation was significantly higher in newborns with high-risk factors of maternal pregnancy. CONCLUSION: The positive rate of deafness gene mutations in the Zhejiang region is higher than that of the database, mainly in GJB2c.235delC, SLC26A4 c.919-2A > G, and m.1555A > G variants. The expanded genetic screening in the detection rate of diseasecausing variants was significantly improved. It is helpful in identifying high-risk children for follow-up intervention.

Efficacy of the combined use of donepezil with either quetiapine or sodium valproate in patients with Alzheimer's disease with behavioral and psychological symptoms of dementia, and their effects on vascular endothelial growth factors.

The present study aimed to compare the clinical efficacy of donepezil combined with quetiapine and with sodium valproate on behavioral and psychological symptoms of dementia (BPSD) in patients with Alzheimer's disease (AD), and to explore the changes and clinical value of vascular endothelial growth factor (VEGF). For this purpose, a total of 131 patients with AD admitted to the Infirmary of Shandong Agricultural University from January, 2017 to January, 2019 were included, of which 60 treated with donepezil combined with quetiapine were designated as group A, whereas 71 treated with donepezil combined with sodium valproate were designated as group B. The behavioral pathology in the AD rating scale (BEHAVE-AD) was used for the evaluation of the clinical efficacy, the brief psychiatric rating scale (BPRS) for the mental state assessment, and the mini-mental state examination (MMSE) for the assessment of cognitive performance. Any adverse reactions were recorded, and the treatment costs of the drugs were compared. According to the treatment efficacy, the patients were divided into the excellent efficacy group and the poor efficacy group. No significant differences were observed in clinical efficacy, or in the single and total adverse reactions between the 2 groups (P>0.05). The drug treatment costs in group A were significantly higher than those in group B (P<0.05). The expression of VEGF in the excellent efficacy group was significantly higher than that in the poor efficacy group (P<0.05). VEGF was found to negatively correlate with the BEHAVE-AD score before and after treatment (P<0.05). On the whole, the present study demonstrates that both quetiapine and sodium valproate combined with donepezil are effective in the treatment of patients with AD presenting with BPSD; the latter is relatively more cost-effective and thus may be worthy of clinical promotion. Moreover, VEGF negatively correlates with BEHAVE-AD score and can thus be used as a potential predictive marker for the treatment response of patients AD with BPSD.

Cellular angiofibroma in the hypopharynx: A case report.

RATIONALE: Cellular angiofibroma is a rare benign mesenchymal tumor which mostly occurs in the superficial soft tissues of the genital region. Occurrence in the head and neck region is extremely rare. To our knowledge, this is the first case of cellular angiofibroma arising in the hypopharynx. PATIENT'S CONCERNS: A 54-year-old male complained of a mass moving through his throat for 1 month. A tumor was found in the right lateral wall of the pharyngeal at the level of the epiglottis by laryngoscope. Magnetic resonance imaging confirmed the presence of a mass in the right lateral pharyngeal area. A benign tumor was suspected based on the clinical symptoms and imaging findings. DIAGNOSIS: A supporting laryngoscope was performed under general anesthesia and the lesion was resected. Immunohistochemical analysis revealed cellular angiofibroma. INTERVENTIONS: The patient underwent surgical excision of the lesion. OUTCOMES: Thus far, no recurrence has been observed 6 months after excision. LESSONS: Cellular angiofibroma located in the lateral pharyngeal is rare; however, immunohistochemical staining is helpful for its diagnosis. Treatment is relatively simple and requires local excision and follow-up.

Characterization of B-BOX gene family and their expression profiles under hormonal, abiotic and metal stresses in Poaceae plants.

BACKGROUND: B-box (BBX) proteins play important roles in plant growth regulation and development including photomorphogenesis, photoperiodic regulation of flowering, and responses to biotic and abiotic stresses. RESULTS: In the present study we retrieved total 131 BBX members from five Poaceae species including 36 from maize, 30 from rice, 24 from sorghum, 22 from stiff brome, and 19 from Millet. All the BBX genes were grouped into five subfamilies on the basis of their phylogenetic relationships and structural features. The expression profiles of 12 OsBBX genes in different tissues were evaluated through qRT-PCR, and we found that most rice BBX members showed high expression level in the heading stage compared to seedling and booting stages. The expression of OsBBX1, OsBBX2, OsBBX8, OsBBX19, and OsBBX24 was strongly induced by abiotic stresses such as drought, cold and salt stresses. Furthermore, the expression of OsBBX2, OsBBX7, OsBBX17, OsBBX19, and OsBBX24 genes was up-regulated under GA, SA and MeJA hormones at different time points. Similarly, the transcripts level of OsBBX1, OsBBX7, OsBBX8, OsBBX17, and OsBBX19 genes were significantly affected by heavy metals such as Fe, Ni, Cr and Cd. CONCLUSION: Change in the expression pattern of BBX members in response to abiotic, hormone and heavy metal stresses signifies their potential roles in plant growth and development and in response to multivariate stresses. The findings suggest that BBX genes could be used as potential genetic markers for the plants, particularly in functional analysis and determining their roles under multivariate stresses.

Second branchial cleft anomalies in children: a literature review.

Branchial cleft anomalies are the second most common head and neck congenital lesions in children. It may sometimes be a part of branchio-oto-renal (BOR) syndrome, so in patients with branchial cleft anomalies associated with a complaint of auricular deformity or a similar history and findings in other family members, we should take an additional examination to find the possibility of BOR syndrome. Complete excision is essential for good prognosis. For the management of branchial cleft anomalies, various methods have been reported. Endoscopically assisted dissection technique and transoral robot-assisted surgery were used in the management of fistula and allowed excellent visualization of the pharyngeal component of the lesion and a minimally invasive approach. It is essential for the surgeon to fully comprehend the congenital lesions to attain the correct preoperative diagnosis and plan for an appropriate surgical approach to prevent the most common complication and recurrence in these lesions. The following sections discuss the anatomy, common presentation, auxiliary examination, differential diagnosis, the current principles of surgical treatment and prognosis for second branchial cleft anomalies in children, and discussed the branchio-oto-renal syndrome.

LMO4 Is a Disease-Provocative Transcription Coregulator Activated by IL-23 in Psoriatic Keratinocytes.

Psoriasis is an autoimmune disease characterized by abnormal differentiation and hyperproliferation of epidermal keratinocytes. LIM-domain only protein 4 (LMO4) is a transcription factor coregulator that promotes the assembly of multiprotein complexes to regulate mammary epithelium and keratinocyte differentiation and proliferation during embryogenesis. In this study, LMO4 has been found to be abundantly expressed in psoriatic epidermis. LMO4 expression is increased in human keratinocytes induced to differentiate by calcium ex vivo, and LMO4 overexpression induces spontaneous differentiation and growth acceleration of human keratinocytes in the absence of calcium. IL-23, a cytokine highly expressed in psoriatic skin lesions, induces differentiation and promotes proliferation of human keratinocytes. The IL-23-mediated effects are accompanied by an increase in LMO4 expression mediated by signal transducer and activator of transcription 3 through an IL-23/acutely transforming retrovirus AKT8 in rodent T-cell lymphoma/signal transducer and activator of transcription 3 pathway in keratinocytes. Knockdown of LMO4 effectively inhibits differentiation and growth of keratinocytes both ex vivo and in IL-23-injected ears of mice. LMO4 appears to mediate IL-23-related responses in psoriatic keratinocytes and is a potential therapeutic target in psoriasis.

PKCδ stabilizes TAp63 to promote cell apoptosis.

PKCδ and p63 are respectively reported to play important roles in cell apoptosis. But there is no report on interaction between them in regulation of apoptosis. In the present study, we found that PKCδ can directly associate and up-regulate TA isoforms of p63 (TAp63) proteins via increasing their stability. PKCδ kinase activity and Thr157 site in TAp63 are crucial for this PKCδ-induced accumulation of TAp63. PKCδ can also enhance TAp63-mediated transcription and cell apoptosis. Taken together, our data indicate that PKCδ phosphorylates TAp63 proteins at Thr157 to stabilize them and promote cell apoptosis.

MUC1 activates JNK1 and inhibits apoptosis under genotoxic stress.

The MUC1 transmembrane glycoprotein is aberrantly overexpressed in diverse human carcinomas and has been shown to inhibit apoptosis induced by genotoxic agents. In the present work, we report that MUC1 binds to and activates JNK1, an important member of the mitogen-activated protein kinases (MAPK) superfamily. The physical interaction between MUC1 cytoplasmic domain (MUC1-CD) and JNK1 was established by GST-pull-down assay in vitro and co-immunoprecipitation assay in vivo. We show that MUC1 activates JNK1 and inhibits cisplatin-induced apoptosis in human colon cancer HCT116 cells. Pharmacological inhibition of JNK or knockdown of JNK significantly reduces the ability of MUC1 to inhibit cisplatin-induced apoptosis. Together, our data indicate that MUC1 can inhibit apoptosis via activating JNK1 pathway in response to genotoxic anticancer agents.