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Various food-derived bioactive peptides have been found with potential anti-inflammatory effects.

Clinical evidence has established that concomitant traumatic brain injury (TBI) accelerates bone healing, but the underlying mechanism is unclear. This study shows that after TBI, injured neurons, mainly those in the hippocampus, release osteogenic microRNA (miRNA)-enriched small extracellular vesicles (sEVs), which targeted osteoprogenitors in bone to stimulate bone formation. We show that miR-328a-3p and miR-150-5p, enriched in the sEVs after TBI, promote osteogenesis by directly targeting the 3′UTR of FOXO4 or CBL, respectively, and hydrogel carrying miR-328a-3p-containing sEVs efficiently repaires bone defects in rats. Importantly, increased fibronectin expression on sEVs surface contributes to targeting of osteoprogenitors in bone by TBI sEVs, thereby implying that modification of the sEVs surface fibronectin could be used in bone-targeted drug delivery. Together, our work unveils a role of central regulation in bone formation and a clear link between injured neurons and osteogenitors, both in animals and clinical settings., Concomitant traumatic brain injury accelerates bone healing, but the mechanism is unclear. Here, the authors show that injured neurons, mainly those in the hippocampus, release osteogenic miRNA-enriched small extracellular vesicles, which targete osteoprogenitors to stimulate bone formation.

The rapid dissemination of antibiotic resistance accelerates the desire for new antibacterial agents. Here, a class of antimicrobial peptides (AMPs) is designed by modifying the structural parameters of a natural chickpea-derived AMP-Leg2, termed "functionalized chickpea-derived Leg2 antimicrobial peptides" (FCLAPs). Among the FCLAPs, KTA and KTR show superior antibacterial efficacy against the foodborne pathogen Escherichia coli (E. coli) O157:H7 (with MICs in the range of 2.5-4.7 µmol L

Ras homologue enriched in brain 1 (Rheb1), an upstream activator of the mechanistic target of rapamycin complex 1 (mTORC1), is known to modulate various cellular processes. However, its impact on bone metabolism

Vortex ropes occurred in the draft tube when hydraulic turbines operating at off-design conditions, which can generate pressure fluctuations. Cavitation vortex rope is one of the most harmful factors to the safety of hydroturbines which may induce further pressure fluctuations. A study of the multiphase flow in a model turbine is presented in this paper using the software ANSYS CFX. The main emphasis is spending on understanding the mechanism of cavitation evolution and underlying its correlation mechanism with flow instabilities. The results indicate that two types of pressure variations can be captured with a cavitation rope: 1) The type due to the vortex rope rotating whose frequency is 1/5–1/4 times runner rotating frequency; 2) The type due to cavitation volume surge, whose frequency is less than that of vortex rope rotating. The frequency of pressure fluctuation due to cavitation keep contanst as the cavitation number decreases, while the amplitude much increases. While the frequency and amplitude of the pressure variation caused by votex rope rotating increase a little. Based on vorticity transport equation, the vortex and cavitation have a close relation. Cavitation increases the vortex production as well as the pressure fluctuation frequency caused by the rotation of vortex rope.

Tuberous sclerosis complex 1 (Tsc1) is a tumor suppressor that functions together with Tsc2 to negatively regulate the mechanistic target of rapamycin complex 1 (mTORC1) activity. Here, we show that Tsc1 has a critical role in the tight junction (TJ) formation of epithelium, independent of its role in Tsc2 and mTORC1 regulation. When an epithelial cell establishes contact with neighboring cells, Tsc1, but not Tsc2, migrates from the cytoplasm to junctional membranes, in which it binds myosin 6 to anchor the perijunctional actin cytoskeleton to β-catenin and ZO-1. In its absence, perijunctional actin cytoskeleton fails to form. In mice, intestine-specific or inducible, whole-body Tsc1 ablation disrupts adherens junction/TJ structures in intestine or skin epithelia, respectively, causing Crohn's disease-like symptoms in the intestine or psoriasis-like phenotypes on the skin. In patients with Crohn's disease or psoriasis, junctional Tsc1 levels in epithelial tissues are markedly reduced, concomitant with the TJ structure impairment, suggesting that Tsc1 deficiency may underlie TJ-related diseases. These findings establish an essential role of Tsc1 in the formation of cell junctions and underpin its association with TJ-related human diseases.

The evolutionarily conserved mechanistic target of rapamycin (mTOR) forms two functionally distinct complexes, -the mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2)-which differ in their subunit composition. Although the function of mTORC1 has been studied extensively, the interaction between mTORC1 and the ubiquitin-proteasome system (UPS) remains unclear. To facilitate a thorough understanding of the mechanismby which UPS regulates mTORC1 activity, steady isotope labeling with amino acids in cell culture (SILAC) technology was used to screen for potential mTORC1-interacting UPS members. Fourteen previously unknown proteins bound to mTOR in HEK293 cells with a SILAC ratio (heavy/light, H/L) above 2, five of which are components of the UPS. Subsequent immunoprecipitation analysis confirmed that ubiquitin-relevant protein 2-like (UBAP2L, also known as NICE-4) binds to both mTOR and Raptor, but not Rictor, suggesting that NICE-4 specifically interacts with mTORC1, but not mTORC2. Interestingly, NICE-4 is essential for basic mTORC1 activity in both HeLa cancer cells and HEK293 cells. In addition, NICE-4 depletion markedly suppressed proliferation of both HeLa and HEK293 cells as well as survival of HeLa cells. Collectively, these results revealed the identity of novel mTOR-interacting UPS proteins and established NICE-4 as a critical UPS member that maintains mTORC1 activity.

Osteocytes are the most abundant (90% to 95%) cells in bone and have emerged as an important regulator of hematopoiesis, but their role in neutrophil development and the underlying mechanisms remain unclear. Interleukin 19 (IL-19) produced predominantly by osteocytes stimulated granulopoiesis and neutrophil formation, which stimulated IL-19 receptor (IL-20Rβ)/Stat3 signaling in neutrophil progenitors to promote their expansion and neutrophil formation. Mice with constitutive activation of mechanistic target of rapamycin complex (mTORC1) signaling in osteocytes (Dmp1-Cre) exhibited a dramatic increase in IL-19 production and promyelocyte/myelocytic expansion, whereas mTORC1 inactivation in osteocytes reduced IL-19 production and neutrophil numbers in mice. We showed that IL-19 administration stimulated neutrophil development, whereas neutralizing endogenous IL-19 or depletion of its receptor inhibited the process. Importantly, low-dose IL-19 reversed chemotherapy, irradiation, or chloramphenicol-induced neutropenia in mice more efficiently than granulocyte colony-stimulating factor. This evidence indicated that IL-19 was an essential regulator of neutrophil development and a potent cytokine for neutropenia treatment.

Endoplasmic reticulum (ER) stress has been shown to promote chondrocyte apoptosis and osteoarthritis (OA) progression, but the precise mechanisms via which ER stress is modulated in OA remain unclear. Here we report that DEP domain-containing mTOR-interacting protein (DEPTOR) negatively regulated ER stress and OA development independent of mTOR signaling. DEPTOR is ubiquitinated in articular chondrocytes and its expression is markedly reduced along with OA progression. Deletion of DEPTOR in chondrocytes significantly promoted destabilized medial meniscus (DMM) surgery-induced OA development, whereas intra-articular injection of lentivirus-expressing DEPTOR delayed OA progression in mice. Proteomics analysis revealed that DEPTOR interplayed with TRC8, which promoted TRC8 auto-ubiquitination and degraded by the ubiquitin-proteasome system (UPS) in chondrocytes. Loss of DEPTOR led to TRC8 accumulation and excessive ER stress, with subsequent chondrocyte apoptosis and OA progression. Importantly, an inhibitor of ER stress eliminated chondrocyte DEPTOR deletion-exacerbated OA in mice. Together, these findings establish a novel mechanism essential for OA pathogenesis, where decreasing DEPTOR in chondrocytes during OA progression relieves the auto-ubiquitination of TRC8, resulting in TRC8 accumulation, excessive ER stress, and OA progression. Targeting this pathway has promising therapeutic potential for OA treatment. © 2020 American Society for Bone and Mineral Research (ASBMR).

The functional role of colonic epithelium in the pathogenesis of ulcerative colitis (UC) remains unclear. Here, we reveal a novel mechanism by which colonic epithelia recruit T helper-17 (Th17) cells during the onset of UC. mTOR complex 1 (mTORC1) was hyper-activated in colonic epithelia of UC mice. While colonic epithelial TSC1 (mTORC1 negative regulator) disruption induced constitutive mTORC1 activation in the colon epithelia and aggravated UC, RPTOR (essential mTORC1 component) depletion inactivated mTORC1 and ameliorated UC. TSC1 deficiency enhanced, whereas RPTOR ablation reduced the expression of cyclooxygenase 2 (COX-2), interleukin-1 (IL-1), IL-6, and IL-23, as well as Th17 infiltration in the colon. Importantly, inhibition of COX-2 reversed the elevation in the expression of these proinflammatory mediators induced by TSC1 deficiency, and subsequently reduced the symptoms and pathological characteristics of UC in mouse models. Mechanistically, mTORC1 activates COX-2 transcription via phosphorylating STAT3 and enhancing it's binding to the COX-2 promoter. Consistently, enhanced mTORC1 activity and COX2 expression, as well as strong positive correlation between each other, were observed in colonic epithelial tissues of UC patients. Collectively, our study demonstrates an essential role of epithelial mTORC1 in UC pathogenesis and establishes a novel link between colonic epithelium, Th17 responses, and UC development.

DEP domain containing mTOR-interacting protein (DEPTOR) was originally identified as an in vivo dual inhibitor of mechanistic target of rapamycin (mTOR). It was recently reported to be involved in renal physiology and pathology in vitro; however, its detailed roles and mechanisms in vivo are completely unknown. We observed that DEPTOR expression in the kidney was markedly increased on day 3 after cisplatin treatment, at which time cell apoptosis peaked, implicating DEPTOR in cisplatin-induced acute kidney injury (AKI). We then used the Cre–LoxP system to generate mutant mice in which the DEPTOR gene was specifically deleted in the proximal tubule cells. DEPTOR deficiency did not alter the renal histology or functions in the saline-treated group, indicating that DEPTOR is not essential for kidney function under physiological conditions. Interestingly, DEPTOR deletion extensively preserved the renal histology and maintained the kidney functions after cisplatin treatment, suggesting that the absence of DEPTOR ameliorates cisplatin-induced AKI. Mechanistically, DEPTOR modulated p38 MAPK signaling and TNFα production in vivo and in vitro, rather than mTOR signaling, thus moderating the inflammatory response and cell apoptosis induced by cisplatin. Collectively, our findings demonstrate the roles and mechanisms of DEPTOR in the regulation of the renal physiology and pathology, and demonstrate that the loss of DEPTOR in the proximal tubules protects against cisplatin-induced AKI.

Osteoblasts provide a microenvironmental niche for B-cell commitment and maturation in the bone marrow (BM). Any abnormity of osteoblasts function may result in the defect of B lymphopoiesis. Signaling from mechanistic target of rapamycin complex 1 (mTORC1) has been implicated in regulating the expansion and differentiation of osteoblasts. Thus, we raise a hypothesis that mTORC1 signaling in osteoblasts plays a vital role in B-cell development. Inactivation of mTORC1 in osterix-expressing cells (mainly osteoblast lineage) through Osx-Cre-directed deletion of Raptor (an mTORC1-specific component) resulted in a reduction in the total B-cell population in the BM, which was due to a block in early B-cell development from the pro-B to pre-B cell stage. Further mechanistic studies revealed that this defect was the result of reduction of interleukin-7 (IL-7) expression in osterix-expressing immature osteoblasts, which caused the abnormality of IL-7/Stat5 signaling in early B lymphocytes, leading to an increased apoptosis of pre-B plus immature B cells. In vitro and in vivo studies demonstrated that the addition of exogenous IL-7 partially restored B lymphopoiesis in the BM of Raptor mutant mice. Furthermore, total BM cells cultured in conditioned media from Raptor null immature osteoblasts or media with anti-IL-7 neutralizing antibody failed to differentiate into pre-B and immature B cells, indicating that inactivation of mTORC1 in immature osteoblast cannot fully support normal B-cell development. Taken together, these findings demonstrate a novel role for mTORC1 in the regulation of bone marrow environments that support B-cell differentiation via regulating IL-7 expression. © 2017 American Society for Bone and Mineral Research.

Lysosomes are the recycling center and nutrient signaling hub of the cell. Here, we show that lysosomes also control mesenchymal stem cell (MSC) differentiation by proteomic reprogramming. The chaperone-mediated autophagy (CMA) lysosome subgroup promotes osteogenesis, while suppressing adipogenesis, by selectively removing osteogenesis-deterring factors, especially master transcriptional factors, such as adipogenic TLE3, ZNF423, and chondrogenic SOX9. The activity of the CMA-committed lysosomes in MSCs are controlled by Van-Gogh-like 2 (Vangl2) at lysosomes. Vangl2 directly binds to lysosome-associated membrane protein 2A (LAMP-2A) and targets it for degradation. MSC-specific Vangl2 ablation in mice increases LAMP-2A expression and CMA-lysosome numbers, promoting bone formation while reducing marrow fat. The Vangl2:LAMP-2A ratio in MSCs correlates inversely with the capacity of the cells for osteoblastic differentiation in humans and mice. These findings demonstrate a critical role for lysosomes in MSC lineage acquisition and establish Vangl2-LAMP-2A signaling as a critical control mechanism.

Severe thrombocytopenia is a significant challenge in patients undergoing myelosuppressive chemotherapy for malignancies. Understanding the biology of platelet-producing megakaryocytes development in the bone marrow microenvironment may facilitate the development of novel therapies to stimulate platelet production and prevent thrombocytopenia. We report here that osteoblasts supported megakaryopoiesis by secreting interleukin-9 (IL-9), which stimulated IL-9 receptor (IL-9R)/Stat3 signaling in promoting megakaryopoiesis. IL-9 production in osteoblasts was negatively regulated by the mechanistic target of rapamycin complex 1 (mTORC1) signaling in a NF-κB-dependent manner. Constitutive activation of mTORC1 inhibited IL-9 production in osteoblasts and suppressed megakaryocytic cells expansion, whereas mTORC1 inactivation increased IL-9 production and enhanced megakaryocyte and platelet numbers in mice. In mouse models, we showed that IL-9 administration stimulated megakaryopoiesis, whereas neutralizing endogenous IL-9 or IL-9R depletion inhibited the process. Importantly, we found that low doses of IL-9 efficiently prevented chemotherapy-induced thrombocytopenia (CIT) and accelerated platelet recovery after CIT. These data indicate that IL-9 is an essential regulator of megakaryopoiesis and a promising therapeutic agent for treatment of thrombocytopenia such as CIT.

Astrocytes respond to CNS insults through reactive astrogliosis, but the underlying mechanisms are unclear. In this study, we show that inactivation of mechanistic target of rapamycin complex (mTORC1) signaling in postnatal neurons induces reactive astrogliosis in mice. Ablation of Raptor (an mTORC1-specific component) in postmitotic neurons abolished mTORC1 activity and produced neurons with smaller soma and fewer dendrites, resulting in microcephaly and aberrant behavior in adult mice. Interestingly, extensive astrogliosis without significant astrocyte proliferation and glial scar formation was observed in these mice. The inhibition of neuronal mTORC1 may activate astrogliosis by reducing neuron-derived fibroblast growth factor 2 (FGF-2), which might trigger FGF receptor signaling in astrocytes to maintain their nonreactive state, and FGF-2 injection successfully prevented astrogliosis in Raptor knock-out mice. This study demonstrates that neuronal mTORC1 inhibits reactive astrogliosis and plays an important role in CNS pathologies.

Bioactive peptides from plant protein sources have been continuously identified as nutrient supplements for low toxicity but multiple physiological activities such as antihypertensive, antioxidant, and anti-inflammatory. In this study, wheat bran protein isolate was digested with alcalase to produce wheat bran protein hydrolysate (WPH) that was then separated into different peptide fractions (1, 1-3, 3-5, and 5-10 kDa) by membrane ultrafiltration. WPH and the peptide fractions were evaluated for in vitro activities such as antioxidant, renin inhibition, and angiotensin-converting enzyme (ACE) inhibition. In addition, the blood pressure-lowering effects of WPH and the 1 kDa peptides were determined by oral administration to spontaneously hypertensive rats (SHRs). Results showed that the ACE and renin inhibitions were significantly (p .05) higher for the 1 kDa fraction (84.25% ± 2.45%, 75.19% ± 1.75%, respectively) when compared to the WPH and 1 kDa fractions. The 1 kDa fraction also showed significantly (p .05) higher oxygen radical antioxidant activity with 2044.73 ± 37.45 (μM TE/g protein) when compared to lower values obtained for the 1 kDa membrane fractions and WPH. Oral administration (100 mg/kg body weight) of the 1 kDa membrane fraction to SHRs resulted in a better decrease (-35 mmHg) in the systolic blood pressure when compared to the WPH (-20 mmHg) after 6 hr. And seven peptides (NL, QL, FL, HAL, AAVL, AKTVF, and TPLTR) were identified and amino acid sequence was determined by tandem mass spectrometry. We conclude that the WPH could be considered as a suitable natural antihypertensive and antioxidant resource. PRACTICAL APPLICATIONS: The results of the present study indicate that WPH and its ultrafiltration fractions possess potential as a source of antihypertensive and strong antioxidant peptides. It has been proved that wheat bran has a good blood pressure lowering and antioxidation and other biological activities, and the 1 kDa fraction showing high oxygen radical absorbance capacity level also has better in vitro ACE inhibition and renin-inhibitory activity. The higher antihypertensive efficiency of the 1 kDa fraction may be because the peptides can be better absorbed from the gastrointestinal tract or an increased ability to interact with the enzyme (ACE or renin) protein structure to change the active conformation and lead to decreased catalysis. The results of this study indicate that WPH, especially 1 kDa peptide, can be used as a component in formulating antihypertensive functional foods and nutraceuticals, thus improving the industrial production efficiency and bioavailability of wheat bran.

To provide theoretical basis for cavitation noise control, the cavitation evolution around a hydrofoil and its induced noise were numerically investigated. A modified turbulence model and Zwart cavitation model were employed to calculate the flow field and predict the cavitation phenomenon accurately. Then, the acoustic analogy method based on the Ffowcs Williams-Hawking (FW-H) equation was applied to analyze the cavitation-induced noise. Seven cavitation numbers were selected for analysis. Acoustic power spectral density (PSD) and acoustic pressure were investigated to establish the relationship between cavitation number and their acoustic characteristics. It was indicated that as cavitation number decreases, cavitation cycle length gets shorter and the magnitude of acoustic power spectral density increases dramatically. One peak value of acoustic power spectral density induced by the extending and retracting of leading-edge cavitation can be obtained under sheet cavitation conditions, while under cloud cavitation, two peak values of acoustic power spectral density can be obtained and are induced by superposition from leading-edge cavitation and trailing vortex.

This paper explores and defines a judicial knowledge reasoning model based on representation learning. The judicial judgment documents of our country collect data in xml format, and represent the key information of the case through the form of attribute-value. In the judicial documents, there is a specific relationship between the information. In order to describe this relationship and predict the element values in the xml through this relationship, this paper makes full use of the data characteristics and the theory of representation learning, and uses the judgment document data as the judicial knowledge base. Construct a judicial knowledge reasoning model based on representation learning, and help judicial staff to check for missing gaps and provide reference analysis when writing judgment documents.

Under the blessing of artificial intelligence technology, legal research is moving towards intelligent and automated. The analysis of judicial data through artificial intelligence methods has become an urgent demand. High-efficient use of data in the judiciary can reduce the burden on relevant personnel, improve the case-handling efficiency, enhance trial quality, and promote fair enforcement. In order to solve the problem of automatic sentencing in legal intelligence, this paper proposes a framework based on FastText and TextCNN to combine the deep learning with justice to predict accusation, converting accusation prediction into a multi-label text classification problem. Compared to traditional methods and machine learning models, our framework can achieve excellent performance.

Osteocytes secrete the glycoprotein sclerostin to inhibit bone formation by osteoblasts, but how sclerostin production is regulated in osteocytes remains unclear. Here, we show that tuberous sclerosis complex 1 (TSC1) in osteocytes promotes sclerostin secretion through inhibition of mechanistic target of rapamycin complex 1 (mTORC1) and downregulation of Sirt1. We generated mice with DMP1 -Cre-directed Tsc1 gene deletion ( Tsc1 CKO) to constitutively activate mTORC1 in osteocytes. Although osteocyte TSC1 disruption increased RANKL expression and osteoclast formation, it markedly reduced sclerostin production in bone, resulting in severe osteosclerosis with enhanced bone formation in mice. Knockdown of TSC1 activated mTORC1 and decreased sclerostin, while rapamycin inhibited mTORC1 and increased sclerostin mRNA and protein expression levels in MLO-Y4 osteocyte-like cells. Furthermore, mechanical loading activated mTORC1 and prevented sclerostin expression in osteocytes. Mechanistically, TSC1 promotes sclerostin production and prevents osteogenesis through inhibition of mTORC1 and downregulation of Sirt1, a repressor of the sclerostin gene Sost . Our findings reveal a role of TSC1/mTORC1 signalling in the regulation of osteocyte sclerostin secretion and bone formation in response to mechanical loading in vitro . Targeting TSC1 represents a potential strategy to increase osteogenesis and prevent bone loss-related diseases.

Figure S1. Generation of mice with osteocyte-specific deletion of TSC1. (A) Schematic of deletion of TSC1 by DMP1-Cre-mediated recombination. This mutant carries a "floxed" allele of Tsc1, when combined with a mutant carrying DMP1-Cre recombinase gene, exons 17 and 18 of Tsc1 are deleted in osteocytes. (B) Genotyping the offspring after mating transgenic Cre and loxp mice. Original image of agarose gel electrophoresis. The expected band of mutant carrying a DMP1-Cre-recombinase gene is nearly 800 bp and floxed Tsc1 allele is nearly 200 bp. M, molecular weight marker. DMP1-Tsc1f/f: control mice/DTCL, DMP1+Tsc1f/f: knockout mice/Tsc1 CKO. (C) Osteocyte deletion of TSC1 was confirmed in cortical sections from Rosa-DMP1 mice. Red signal indicates DMP1 expression in osteocytes. Boxed area was magnified on the right. Scale bar, 50 µm (n = 6); Figure S2. General phenotypes of Tsc1 CKO mice. (A) Body length and (B) weight in 10-week-old control and Tsc1 CKO mice. No difference was detected in control and Tsc1 CKO mice (t test, p=0.1235 and p=0.0873, n = 6). ns, no significance. (C-D) Representative micro-computed tomography (micro-CT) 2D images of (C) vertebral column, (D) metaphyseal trabecular bone of 4-, 8-, and 12-week-old control and Tsc1 CKO mice. Scale bar, 1 mm (n = 6).Data represent mean ± SD.* P < 0.05 ** P < 0.01, ***P < 0.001 by Student's t test.; Figure S3 . Thickness of cortical bone in DTCL and Tsc1 CKO mice. (A) H&E staining of cortical bone of femora from 4-, 8- and 12-week-old control and Tsc1 CKO mice. The boxed area was magnified in the panel below. The scale bar represents 100 µm and 50 µm (n = 6). (B-C) Histomorphometric measurements showed that (B) the mineralizing surface/bone surface (MS/BS, t test, p = 0.0087) and (C) bone formation rate (BFR, t test, p = 0.0003) of control mice were lower than that of Tsc1 CKO mice (n = 6). (D) mRNA expression of TSC1 in DTCL and Tsc1 CKO mice (t test, p = 0.0007). Data are represented as mean ± SD, **p < 0.01, and ***p < 0.001.; Figure S4 . TSC1 deletion induced H3K9 acetylation of the Sost promoter. (A)CHIP-qPCR detected TSC1 binding to the Sost promotor region in MLO-Y4 with control or TSC1 shRNA lentivirus infection. (t test, -3602-3728, p = 0.1476; -761-844, p = 0.7348; -998-1115, p = 0.1037; -1245-1361, p = 0.0141;-1828-1940, p = 0.0209; n = 3) .Data are represented as mean ± SD. * p < 0.05.

Antimicrobial food packaging with a sustained release of antimicrobial agents plays a key role in maintaining food quality and food safety. In this study, polyelectrolyte gellan gum (GG)-chitosan (CS) multilayer film was fabricated by layer-by-layer assembly technology with the incorporation of thyme essential oil (TEO) coarse emulsion (TEOC) or nanoemulsion (TEON). The microstructure of GG-CS three-layered film was observed by scanning electron microscopy (SEM). The red fluorescent signals in confocal laser scanning microscopy (CLSM) images indicated smaller droplets and more uniform distribution of TEON prepared by ultrasonic treatments, which was consistent with the particle size results and porous structure in SEM images. The film incorporated with 6% TEON exhibited higher antimicrobial activity than that with 6% TEOC, and it significantly reduced E. coli populations in liquid model system. Moreover, films incorporated with TEON showed improved mechanical flexibility (elongation at break) and UV blocking property in comparison to TEOC. This developed GG-CS three-layered film loading with TEON was expected to be a potential antimicrobial material with sustained release property for food packaging applications.

The evolutionarily conserved mechanistic target of rapamycin (mTOR) forms two functionally distinct complexes, mTORC1 and mTORC2. mTORC1, consisting of mTOR, raptor, and mLST8 (GβL), is sensitive to rapamycin and thought to control autonomous cell growth in response to nutrient availability and growth factors. mTORC2, containing the core components mTOR, mLST8, Rictor, mSIN1, and Protor1/2 is largely insensitive to rapamycin. mTORC2 specifically senses growth factors and regulates cell proliferation, metabolism, actin rearrangement, and survival. Dysregulation of mTOR signaling often occurs in a variety of human malignant diseases, rendering it a crucial and validated target in cancer treatment. However, the effectiveness of rapamycin as single-agent therapy is suppressed, in part, by the numerous strong mTORC1-dependent negative feedback loops. Although preclinical and clinical studies of ATP-competitive mTOR inhibitors that target both mTORC1 and mTORC2 have shown greater effectiveness than rapalogs for cancer treatment, the mTORC1 inhibition-induced negative feedback activation of PI3- K/PDK1 and Akt (Thr308) may be sufficient to promote cell survival. Recent cancer biology studies indicated that mTORC2 is a promising target, since its activity is essential for the development of a number of cancers. These studies provide a rationale for developing inhibitors specifically targeting mTORC2, which do not perturb the mTORC1- dependent negative feedback loops and have a more acceptable therapeutic window. This review summarizes the present understanding of mTORC2 signaling and functions, especially tumorigenic functions, highlighting the current status and future perspectives for targeting mTORC2 in cancer treatment.

Sertoli cells (SCs) play a central role in testis development, and their normal number and functions are required for spermatogenesis. Although the canonical tuberous sclerosis complex-mammalian target of rapamycin complex 1(TSC-mTORC1) pathway is critical for testis development and spermatogenesis, the signaling mechanisms governing SC functions remain unclear. In this study, we generated two SC-specific mouse mutants using the Cre-LoxP system. Loss of Raptor (a key component of mTORC1) caused severe tubular degeneration in the neonatal testis and adult mice displayed azoospermia, while adult Rheb (an upstream activator for mTORC1) mutant mice had intact tubules and many sperm in their epididymides. Disruption of cytoskeletal organization, including actin, microtubules, and SC-intrinsic vimentin, was observed in Raptor but not Rheb mutant mice. We investigated the reasons for these different effects by whole-transcriptome sequencing, and found that expression of the tight junction adaptor protein cingulin was significantly reduced in Raptor mutant mice. The expression profile of cingulin was synchronous with the differentiation and cytoskeletal dynamics of SCs in control mice, but was disordered in Raptor mutant mice. Furthermore, activity of the small GTPase Rac1 was reduced and expression of the guanine exchange factor for Rac1, Asef, was decreased in Raptor but not Rheb mutant mice. Collectively, these findings establish novel functions of Raptor, independent of the canonical Rheb/mTORC1 pathway, in controlling cytoskeletal homeostasis and cell polarity in SCs, by affecting cingulin expression and Rac1 activity.

Osteoporosis is characterized by low bone mineral density and microarchitectural deterioration of bone tissue. N-(3-methoxybenzyl)-(9Z,12Z,15Z)-octadecatrienamide (MBOC) is one of the macamides isolated from Maca (Lepidium meyenii Walp.), a cruciferous plant from the Andes of Peru. In this study, C3H/10T1/2 mesenchymal stem cells were treated with MBOC in osteogenic induction medium. An ovariectomized (OVX) mouse model was used to investigate the effect of 1-month MBOC treatment on the prevention of postmenopausal osteoporosis. Remarkably, trabecular thickness, trabecular number, and bone volume/tissue volume of the distal femoral metaphysis were significantly increased in OVX + MBOC mice compared with OVX mice, as revealed by microcomputed tomography analysis. Trabecular separation was decreased in OVX + MBOC mice compared with OVX mice. Consistently, MBOC increased the levels of osteocalcin and runt-related transcription factor 2 in OVX mice, as well as the expression of runt-related transcription factor 2, osterix, and alkaline phosphatase in C3H/10T1/2 cells. Mechanistically, MBOC activates the canonical Wnt/β-catenin signaling pathway via inhibiting phosphorylation of GSK-3β at Tyr216 and maintaining β-catenin expression. Collectively, the current study demonstrates the robustness of MBOC in the induction of mesenchymal stem cells osteogenic differentiation and consequent bone formation, suggesting that MBOC may be a potentially effective drug to treat postmenopausal osteoporosis.

Osteoblasts provide a microenvironmental niche for B-cell commitment and maturation in the bone marrow (BM). Any abnormity of osteoblasts function may result in the defect of B lymphopoiesis. Signaling from mechanistic target of rapamycin complex 1 (mTORC1) has been implicated in regulating the expansion and differentiation of osteoblasts. Thus, we raise a hypothesis that mTORC1 signaling in osteoblasts plays a vital role in B-cell development. Inactivation of mTORC1 in osterix-expressing cells (mainly osteoblast lineage) through Osx-Cre-directed deletion of Raptor (an mTORC1-specific component) resulted in a reduction in the total B-cell population in the BM, which was due to a block in early B-cell development from the pro-B to pre-B cell stage. Further mechanistic studies revealed that this defect was the result of reduction of interleukin-7 (IL-7) expression in osterix-expressing immature osteoblasts, which caused the abnormality of IL-7/Stat5 signaling in early B lymphocytes, leading to an increased apoptosis of pre-B plus immature B cells. In vitro and in vivo studies demonstrated that the addition of exogenous IL-7 partially restored B lymphopoiesis in the BM of Raptor mutant mice. Furthermore, total BM cells cultured in conditioned media from Raptor null immature osteoblasts or media with anti-IL-7 neutralizing antibody failed to differentiate into pre-B and immature B cells, indicating that inactivation of mTORC1 in immature osteoblast cannot fully support normal B-cell development. Taken together, these findings demonstrate a novel role for mTORC1 in the regulation of bone marrow environments that support B-cell differentiation via regulating IL-7 expression. © 2017 American Society for Bone and Mineral Research.

Exosomes are small membrane‐bound vesicles released into extracellular spaces by many types of cells. These nanovesicles carry proteins, mRNA, and miRNA, and are involved in cell waste management and intercellular communication. In the present study, it is shown that exosome release, which leads to net loss of cellular membrane and protein content, is negatively regulated by mechanistic target of rapamycin complex 1 (mTORC1). It is found that in cells and animal models exosome release is inhibited by sustained activation of mTORC1, leading to intracellular accumulation of CD63‐positive exosome precursors. Inhibition of mTORC1 by rapamycin or nutrient and growth factor deprivation stimulates exosome release, which occurs concomitantly with autophagy. The drug‐stimulated release is blocked by siRNA‐mediated downregulation of small GTPase Rab27A. Analysis of the cargo content in exosomes released from rapamycin‐treated cells reveals that inhibition of mTORC1 does not significantly alter its majority protein and miRNA profiles. These observations demonstrate that exosome release, like autophagy, is negatively regulated by mTORC1 in response to changes in nutrient and growth factor conditions.

// Miao Liu 1, * , Ling Zhou 1, * , Baiyu Zhang 2 , Minhong He 1 , Xiaoying Dong 1 , Xiaojun Lin 3 , Chunhong Jia 3 , Xiaochun Bai 3 , Yifan Dai 4 , Yongchun Su 5 , Zhipeng Zou 3 , Hang Zheng 1 1 Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China 2 Department of Rheumatology, the Sixth Affiliated Hospital of Sun Yat-Sen University, 510655, China 3 Department of Cell Biology, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, China 4 State Key Laboratory of Reproductive Medicine and Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 201129, China 5 Department of Bioinformatics, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, China * These authors contributed equally to this work Correspondence to: Yongchun Su, email: suyc@smu.edu.cn Zhipeng Zou, email: zzp@smu.edu.cn Hang Zheng, email: zhengh001@163.com Keywords: PUFA, mTOR complex 1, colorectal cancer, APC Received: January 23, 2016 Accepted: October 10, 2016 Published: October 19, 2016 ABSTRACT Although epidemiological and preclinical studies have shown the preventative effect of n-3 polyunsaturated fatty acids (PUFAs) on colorectal cancer (CRC), the underlying molecular mechanisms are not clear. In this study, we revealed that elevation of n−3/n-6 PUFAs ratio suppress the mechanistic target of rapamycin complex 1 (mTORC1) and prevent colorectal tumorigenesis. The transgenic expression of fat-1 , a desaturase that catalyzes the conversion of n-6 to n-3 PUFAs and produces n-3 PUFAs endogenously, repressed colorectal tumor cell growth and remarkably reduced tumor burden, and alleviated anemia as well as hyperlipidemia in APC Min/+ (adenomatous polyposis coli) mice, a classic CRC model that best simulates most clinical cases. In contrast to arachidonic acid (AA, C20:4 n−6), either Docosahexaenoic acid (DHA, C22:6 n−3), eicosapentaenoic acid (EPA, C20:5 n−3), or a combination of DHA and AA, efficiently inhibited the proliferation of CRC cell lines and promoted apoptosis in these cells. The ectopic expression of fat-1 had similar effects in colon epithelial cells with APC depletion. Mechanistically, elevation of n−3/n−6 ratio suppressed mTORC1 activity in tumors of APC Min/+ mice, CRC cell lines with APC mutation, and in normal colon epithelial cells with APC depletion. In addition, elevation of n−3/n−6 ratio repressed mTORC1 activity and inhibited adipogenic differentiation in preadipocytes with APC knockdown, as well as alleviated hyperlipidemia in APC Min/+ mice. Taken together, our findings have provided novel insights into the potential mechanism by which increase in n−3/n−6 PUFAs ratio represses CRC development, and also a new rationale for utilizing n-3 PUFAs in CRC prevention and treatment.