Search

Deubiquitinating enzymes (DUBs) play important roles in many physiological and pathological processes by modulating the ubiquitination of their substrates. DUBs undergo post-translational modifications including ubiquitination. However, whether DUBs can reverse their own ubiquitination and regulate their own protein stability requires further investigation. To answer this question, we screened an expression library of DUBs and their enzymatic activity mutants and found that some DUBs regulated their own protein stability in an enzymatic activity- and homomeric interaction-dependent manner. Taking Ubiquitin-specific-processing protease 29 (USP29) as an example, we found that USP29 deubiquitinates itself and protects itself from proteasomal degradation. We also revealed that the N-terminal region of USP29 is critical for its protein stability. Taken together, our work demonstrates that at least some DUBs regulate their own ubiquitination and protein stability. Our findings provide novel molecular insight into the diverse regulation of DUBs.

Regulation of hyaluronan (HA) is important for the maintenance of epidermal homeostasis. Here, we examined the mechanism by which 1-ethyl-β-N-acetylglucosaminide (β-NAG2), a newly developed N-acetylglucosamine (NAG) derivative, increases HA production in cultured human epidermal keratinocytes. When keratinocytes were treated with β-NAG2, mRNA expression of HA synthase 3, which is responsible for HA production in human keratinocytes, was not influenced, but the intracellular level of UDP-NAG, a substrate used for HA synthesis, was increased. By using a synthetic substrate for β-N-acetylglucosaminidase (β-NAGase), keratinocytes were found to possess β-NAGase activity, and treatment of o-(2-acetamido-2-deoxy-d-glucopyranosylidene) amino N-phenyl carbamate (PUGNAc), an inhibitor of β-NAGase, abolished the release of NAG from β-NAG2 in keratinocytes. Furthermore, PUGNAc attenuated the β-NAG2-induced intracellular UDP-NAG and HA production in keratinocytes. These results suggest that β-NAG2 is converted to NAG by endogenous β-NAGase in keratinocytes, and the resulting NAG is further metabolized to UDP-NAG and utilized for HA production.

Hydrogen sulfide (H2S) is a cytoprotective redox-active metabolite that signals through protein persulfidation (R-SSnH). Despite the known importance of persulfidation, tissue-specific persulfidome profiles and their associated functions are not well characterized, specifically under conditions and interventions known to modulate H2S production. We hypothesize that dietary restriction (DR), which increases lifespan and can boost H2S production, expands tissue-specific persulfidomes. Here, we find protein persulfidation enriched in liver, kidney, muscle, and brain but decreased in heart of young and aged male mice under two forms of DR, with DR promoting persulfidation in numerous metabolic and aging-related pathways. Mice lacking cystathionine γ-lyase (CGL) have overall decreased tissue protein persulfidation numbers and fail to functionally augment persulfidomes in response to DR, predominantly in kidney, muscle, and brain. Here, we define tissue- and CGL-dependent persulfidomes and how diet transforms their makeup, underscoring the breadth for DR and H2S to impact biological processes and organismal health., Dietary restriction (DR) can increase protein persulfidation but the tissue specificity of this process is not well understood. Here, the authors compare organ-specific protein persulfidomes in young and aged mice under DR, and show that DR-dependent persulfidome changes depend on cystathionine γ-lyase.

Prunella vulgaris (PV) is a perennial plant which is widely grown around the world. It has been widely used as a medicinal treatment for generations. Previous studies showed extracts from this plant had a wide range of therapeutic efficacy, including anti-tumorous effect. However, the volatile organic compounds (VOCs) extracted from it were rarely explored. This paper reports on the characterization of a steam distillation process to extract VOCs in PV and also the anti-tumorous effects of the PV distillate using the tetrazolium-based Cell Counting Kit-8 (CCK-8) as the test agent, when the VOCs were used to treat oral squamous cancer cells, SSC154. It was found that most abundant VOCs came out steadily and continuously for as long as the duration of the steam extraction could extend. However, some compounds such as benzaldehyde did show depletion as the distillation process progressed, while some compounds such as caryophyllene oxide was only sparsely found at the beginning of distillation. The PV distillate was mildly effective in its cytotoxicity to cancer cells SCC154, in a dosage dependent manner.

Chitosan is a bio-based polysaccharide having promising biological and antitumor properties. The present study aims at the synthesis of chitosan using shrimp shell waste. Chitosan preparation consists of two consecutive steps such as demineralization and deproteinization. The structure, morphology and optical properties of the obtained chitosan were established using various instrumental techniques like XRD, FTIR, UV–Vis, SEM and TEM. Herein, we have shown that Significant bacterial activity was manifested by chitosan against both gram positive (S. aureus and S. mutans) and gram negative (E. coli and P. aeuriginosa) bacteria. The biosynthesized chitosan exerts an inhibitory effect on the proliferation of MCF-7 breast cancer cells in a dose –dependent manner while being non -toxic to fibroblast L929 normal cells.

MicroRNAs (miRNAs) play critical roles in gene expression and numerous human diseases. The success of miRNA biogenesis is largely determined by the primary miRNA (pri-miRNA) processing by the DROSHA-DGCR8 complex, called Microprocessor. Here, we analysed the high-throughput pri-miRNA processing assays and secondary structures of pri-miRNAs to investigate the roles of bulges in the pri-miRNA processing. We found that bulges in multiple places control both the cleavage efficiency and accuracy of pri-miRNA processing. These bulges were shown to act on Microprocessor via its catalytic subunit, DROSHA, and function in a position and strand-dependent manner. Interestingly, we discovered that the enriched and conserved bulges, called midB, can correct DROSHA orientation on pri-miRNAs, thereby enhancing production of miRNAs. The revealed functions of the bulges help improve our understanding of pri-miRNA processing and suggest their potential roles in miRNA biogenesis regulation.

N6-methyladenosine (m6A) RNA methylation is the most prevalent modification of messenger RNAs (mRNAs) and catalyzed by a multicomponent methyltransferase complex (MTC), among which methyltransferase-like 3 (METTL3) and METTL14 are two core molecules. However, METTL3 and METTL14 play opposite regulatory roles in hepatocellular carcinoma (HCC). Based on The Cancer Genome Atlas (TCGA) database and Gene Expression Omnibus (GEO) database, we conducted a multi-omics analysis of METTL3 and METTL14 in HCC, including RNA-sequencing, m6ARIP-sequencing, and ribosome-sequencing profiles. We found that the expression and prognostic value of METTL3 and METTL14 are opposite in HCC. Besides, after METTL3 and METTL14 knockdown, most of the dysregulated mRNAs, signaling pathways and biological processes are distinct in HCC, which partly explains the contrary regulatory role of METTL3 and METTL14. Intriguingly, these mRNAs whose stability or translation efficiency are influenced by METTL3 or METTL14 in an m6A dependent manner, jointly regulate multiple signaling pathways and biological processes, which supports the cooperative role of METTL3 and METTL14 in catalyzing m6A modification. In conclusion, our study further clarified the contradictory role of METTL3 and METTL14 in HCC.

Ebola virus (EBOV) RNA has the potential to form hairpin structures at the transcription start sequence (TSS) and reinitiation sites of internal genes, both on the genomic and antigenomic/mRNA level. Hairpin formation involving the TSS and the spacer sequence between promotor elements (PE) 1 and 2 was suggested to regulate viral transcription. Here, we provide evidence that such RNA structures form during RNA synthesis by the viral polymerase and affect its activity. This was analysed using monocistronic minigenomes carrying hairpin structure variants in the TSS-spacer region that differ in length and stability. Transcription and replication were measured via reporter activity and by qRT-PCR quantification of the distinct viral RNA species. We demonstrate that viral RNA synthesis is remarkably tolerant to spacer extensions of up to ~54 nt, but declines beyond this length limit (~25% residual activity for a 66-nt extension). Minor incremental stabilizations of hairpin structures in the TSS-spacer region and on the mRNA/antigenomic level were found to rapidly abolish viral polymerase activity, which may be exploited for antisense strategies to inhibit viral RNA synthesis. Finally, balanced viral transcription and replication can still occur when any RNA structure formation potential at the TSS is eliminated, provided that hexamer phasing in the promoter region is maintained. Altogether, the findings deepen and refine our insight into structure and length constraints within the EBOV transcription and replication promoter and suggest a remarkable flexibility of the viral polymerase in recognition of PE1 and PE2.

International audience; Phosphorus (P) is an essential macronutrient for all living organisms. Importantly, plants require a large amount of P to grow, and P deficiency causes huge losses in plant production. Although this issue can be mitigated by the appropriate use of phosphate (Pi) rock-derived P fertilizers, phosphate rock is a finite natural resource. Moreover, the increased demand for food as a result of our growing global population is another factor contributing to a prospective P crisis. While creating crops that are resilient to Pi deficiency presents great scientific challenge, the current progress in our understanding of how plants regulate Pi homeostasis offers some opportunities for further study. In this review, we present the published research supporting these opportunities, which are based on the molecular mechanisms that plants have evolved to respond to P deficiency. First, we focus on recent advances in P sensing and signaling pathways in the regulation of root system architecture. Next, we describe the mechanisms that regulate Pi transport and accumulation, in a Pi- (or other nutrient) dependent manner. Integrating these data will help to design an innovative strategy for improving Pi nutrition in plants. In addition, this will help with Pi scarcity, one of the challenges facing agriculture in the twenty first century.

Previously, it was found that the hepatitis C virus NS5A interacted with ACBD3 in a genotype-dependent manner. However, the region in NS5A responsible for association with ACBD3 is not clear. Domain I of NS5A was identified as critical for ACBD3 binding. By comparing the differences of amino acids in domain I from different genotypes of NS5A, it was found that key amino acids potentially corresponded to the affinity of the NS5A-ACBD3 interaction. The findings not only revealed that domain I of NS5A associates with ACBD3 but they also shed mechanistic light on how NS5A is associated with ACBD3 in a genotype-dependent manner.

Natural antiangiogenic products interact with multiple molecular angiogenic pathways and possesses comparably less toxicities then chemically synthesized anti-angiogenic compounds. There is a need to screen varieties of medicinal plants to identify more sources of antiangiogenic compounds, as anticancer drugs with less side effects. This study was designed to explore the antiangiogenic property and cytotoxicity of Moricandia sinaica (Boiss.) in zebrafish embryos, as well as its anti-migratory potential in human umbilical vein endothelial cells (HUVECs). Gas chromatography coupled with mass spectrophotometry was performed in order to identify bioactive molecules present in the stem and leaves part of Moricandia sinaica.The methanol extract of the stem and leaves inhibited the formation of the inter-segmental, sub-intestinal vein, and dorsal longitudinal anastomotic veins formation in zebrafish embryos in a dose- dependent manner. It did not show cytotoxicity at lower concentration in HUVECs; however, it significantly (p

Background Several findings suggest neuroinflammation as a contributing factor for the onset of psychiatric disorders such as Alzheimer’s disease, depression, and anxiety. There is increasing evidence pointing out that the Mediterranean diet influences brain and behavior. Mediterranean herbs and spices have been shown to be within those components of the Mediterranean diet involved in cognitive enhancement. Thus, we investigated the influence of Mediterranean natural extracts (MNE), Rosemary extract (RE) and Glycyrrhiza glabra root extract (GGRE), on cognitive behavior. Results Adult zebrafish were exposed to RE or GGRE (100 and 250 mg/L) treatments. Both MNE improved memory retention during the T-maze test, although no improvements were observed during the novel object preference. Similarly, chronic administration of RE (150 mg/Kg) and GGRE (150 mg/Kg) improved, respectively, spatial and retention memory, as assessed by the Morris Water Maze (MWM), and the Elevated Plus Maze (EPM) in healthy male rats. However, no improvements were observed during the novel object recognition. Finally, male, and female rats were chronically treated with lipopolysaccharide [(LPS) 300 ug/kg] and orally administered with RE. Interestingly, RE reversed LPS-induced cognitive deficit during the MWM and EPM in female rats. Conclusions We found that MNE improved cognition in both zebrafish and rats. Moreover, MNE rescued LPS-induced cognitive impairment in a gender-specific manner. Therefore, our study supports the view that zebrafish represent a valuable preclinical model for drug discovery in neuroscience. These findings contribute to an exciting and growing body of research suggesting that MNE may play an important role in the prevention of cognitive impairment.

Heat activation at a sublethal temperature is widely applied to promote Bacillus species spore germination. This treatment also has the potential to be employed in food processing to eliminate undesired bacterial spores by enhancing their germination and then inactivating the less-heat-resistant germinated spores at a milder temperature. However, incorrect heat treatment could also generate heat damage in spores and lead to more heterogeneous spore germination. Here, the heat activation and heat damage profile of Bacillus subtilis spores was determined by testing spore germination and outgrowth at both population and single-spore levels. The heat treatments used were 40 to 80°C and for 0 to 300 min. The results were as follows. (i) Heat activation at 40 to 70°C promoted l-valine- and l-asparagine-glucose-fructose-potassium (AGFK)-induced germination in a time-dependent manner. (ii) The optimal heat activation temperatures for AGFK and l-valine germination via the GerB plus GerK or GerA germinant receptors were 65°C and 50 to 65°C, respectively. (iii) Heat inactivation of dormant spores appeared at 70°C, and the heat damage of molecules essential for germination and growth began at 70 and 65°C, respectively. (iv) Heat treatment at 75°C resulted in both activation of germination and damage to the germination apparatus, and 80°C treatment caused more pronounced heat damage. (v) For the spores that should withstand adverse environmental temperatures in nature, heat activation seemed functional for a subsequent optimal germination process, while heat damage affected both germination and outgrowth. IMPORTANCE Bacterial spores are thermal-stress-resistant structures that can thus survive food preservation strategies and revive through the process of spore germination. The more heat resistant spores are, the more heterogeneous their germination upon the addition of germinants. Upon germination, spores can cause food spoilage and food intoxication. Here, we provide new information on both heat activation and inactivation regimes and their effects on the (heterogeneity of) spore germination.

Vif is a lentiviral accessory protein that counteracts the antiviral activity of cellular APOBEC3 (A3) cytidine deaminases in infected cells. The exact contribution of each member of the A3 family for the restriction of HIV-2 is still unclear. Thus, the aim of this work was to identify the A3s with anti-HIV-2 activity and compare their restriction potential for HIV-2 and HIV-1. We found that A3G is a strong restriction factor of both types of viruses and A3C restricts neither HIV-1 nor HIV-2. Importantly, A3B exhibited potent antiviral activity against HIV-2, but its effect was negligible against HIV-1. Whereas A3B is packaged with similar efficiency into both viruses in the absence of Vif, HIV-2 and HIV-1 differ in their sensitivity to A3B. HIV-2 Vif targets A3B by reducing its cellular levels and inhibiting its packaging into virions, whereas HIV-1 Vif did not evolve to antagonize A3B. Our observations support the hypothesis that during wild-type HIV-1 and HIV-2 infections, both viruses are able to replicate in host cells expressing A3B but using different mechanisms, probably resulting from a Vif functional adaptation over evolutionary time. Our findings provide new insights into the differences between Vif protein and their cellular partners in the two human viruses. Of note, A3B is highly expressed in some cancer cells and may cause deamination-induced mutations in these cancers. Thus, A3B may represent an important therapeutic target. As such, the ability of HIV-2 Vif to induce A3B degradation could be an effective tool for cancer therapy. IMPORTANCE Primate lentiviruses encode a series of accessory genes that facilitate virus adaptation to its host. Among those, the vif-encoded protein functions primarily by targeting the APOBEC3 (A3) family of cytidine deaminases. All lentiviral Vif proteins have the ability to antagonize A3G; however, antagonizing other members of the A3 family is variable. Here, we report that HIV-2 Vif, unlike HIV-1 Vif, can induce degradation of A3B. Consequently, HIV-2 Vif but not HIV-1 Vif can inhibit the packaging of A3B. Interestingly, while A3B is packaged efficiently into the core of both HIV-1 and HIV-2 virions in the absence of Vif, it only affects the infectivity of HIV-2 particles. Thus, HIV-1 and HIV-2 have evolved two distinct mechanisms to antagonize the antiviral activity of A3B. Aside from its antiviral activity, A3B has been associated with mutations in some cancers. Degradation of A3B by HIV-2 Vif may be useful for cancer therapies.

SummaryDeveloping methods to modulate growth of the mucin-degrading gut bacterium Akkermansia muciniphila could benefit patients with different health needs, as A. muciniphila has been associated with both positive metabolic health outcomes and detrimental neurodegenerative outcomes. Growth of A. muciniphila is sensitive to plant-derived polyphenols, and particularly proanthocyanidins (PACs), when administered in isolated form at supraphysiological doses. However, it remains unclear whether doses sufficient for these effects are achievable via diet. Here, we explore the extent to which nutritionally relevant doses of common polyphenol-rich foods – berries, wine, and coffee – influence A. muciniphila abundance in C57BL/6J mice under varying dietary conditions. By administering polyphenol-rich whole foods, comparing polyphenol-depleted and PAC-rich versus PAC-poor food supplements, and through gradient PAC-dosing experiments, we show that PAC-rich foods uniquely induce A. muciniphila growth at doses that are feasibly achieved through routine diet. Notably, the effects of PAC supplementation were detected against a high-fat diet but not a low-fat control diet background, highlighting the importance of habitual diet strategies in either amplifying or mitigating the prebiotic effects of PAC-rich food consumption. Ultimately, our work suggests that both PACs and diet influence A. muciniphila abundance with downstream impacts for human health.

The aberrant expression of fat mass and obesity-associated protein (FTO) has been confirmed to be associated with a variety of cancers and participates in the regulation of multiple biological behaviours. FTO plays an oncogenic role in bladder cancer, but few studies have focused on how FTO promotes bladder cancer progression by regulating miRNA synthesis. Here, we confirmed that FTO expression was significantly increased in bladder cancer and was associated with a poor prognosis. FTO overexpression promoted bladder cancer cell proliferation, whereas FTO knockdown inhibited bladder cancer cell proliferation. We also demonstrated that FTO promoted bladder cancer cell proliferation via the FTO/miR-576/CDK6 pathways. Taken together, our work revealed that FTO plays a critical role in bladder cancer and could be a potential diagnostic or prognostic biomarker for this disease.

Background: Enterobacter cloacae (EC) is a commonly occurring opportunistic pathogen and is responsible for causing various infections in humans. Owing to its inducible chromosomal AmpC β-lactamase (AmpC), EC is inherently resistant to the 1st- and 2nd- generation cephalosporins. However, whether β-lactams antibiotics enhance EC resistance remains unclear.Results: In this study, we found that subinhibitory concentrations (SICs) of cefazolin (CFZ) and imipenem (IMP) are able to advance the expression of AmpC and improve its resistance towards β - lactams through NagZ in EC clinical isolate. Our work indicate that AmpC manifested a substantial upregulation in EC in response to SICs of CFZ and IMP. In nagZ knockout EC (ΔnagZ), we found that the resistance to β - lactam antibiotics was rather weakened and the effect of CFZ and IMP on induction of AmpC was completely abrogated. Ectopic expression of NagZ can rescue the induction effect of CFZ and IMP on AmpC and enhance resistance in ΔnagZ. More importantly, CFZ and IMP have the potential to bring about the target genes expressions of AmpR in a NagZ-dependent manner.Conclusions: Our findings show that NagZ is a critical determinant for CFZ and IMP to promote AmpC expression and improve resistance and that CFZ and IMP should be used with caution since they may aggravate EC resistance. At the same time, this study further improves our understanding of resistance mechanisms in EC.

The naïve epiblast transitions to a pluripotent primed state during embryo implantation. Despite the relevance of the FGF pathway during this period, little is known about the downstream effectors regulating this signaling. Here, we examined the molecular mechanisms coordinating the naïve to primed transition by using inducible ESC to genetically eliminate all RAS proteins. We show that differentiated RAS

The spiking activity of populations of cortical neurons is well described by the dynamics of a small number of population-wide covariance patterns, whose activation we refer to as ‘latent dynamics’. These latent dynamics are largely driven by the same correlated synaptic currents across the circuit that determine the generation of local field potentials (LFPs). Yet, the relationship between latent dynamics and LFPs remains largely unexplored. Here, we characterised this relationship for three different regions of primate sensorimotor cortex during reaching. The correlation between latent dynamics and LFPs was frequency-dependent and varied across regions. However, for any given region, this relationship remained stable throughout the behaviour: in each of primary motor and premotor cortices, the LFP-latent dynamics correlation profile was remarkably similar between movement planning and execution. These robust associations between LFPs and neural population latent dynamics help bridge the wealth of studies reporting neural correlates of behaviour using either type of recordings.

SUMMARYThe last stage of cell division involves two daughter cells remaining interconnected by a cytokinetic bridge that is cleaved in a process called abscission. During pre-abscission, we identified that the centrosome moves in a Rab11-dependent manner towards the cytokinetic bridge in human cells grown in culture and in an in vivo vertebrate model, Danio rerio (zebrafish). Rab11-endosomes are dynamically organized in a Rab11-GTP dependent manner at the centrosome during pre-abscission and this organization is required for the centrosome protein, pericentrin, to be enriched at the centrosome. Using zebrafish embryos, we found that reduction in pericentrin expression or optogenetically disrupting Rab11-endosome function inhibited centrosome movement towards the cytokinetic bridge and abscission resulting in daughter cells prone to being binucleated and/or having supernumerary centrosomes. These studies suggest that Rab11-endosomes contribute to centrosome function during pre-abscission by regulating pericentrin organization resulting in appropriate centrosome movement towards the cytokinetic bridge and subsequent abscission.

Pseudomonas aeruginosa, a major cause of nosocomial infection, can survive under diverse environmental conditions. Its great adaptive ability is dependent on its multiple signaling systems such as the two-component system (TCS). A TCS FleS/FleR has been previously identified to positively regulate a variety of virulence-related traits in P. aeruginosa PAO1 including motility and biofilm formation which are involved in the acute and chronic infections, respectively. However, the molecular mechanisms underlying these regulations are still unclear. In this study, we first analyzed the regulatory roles of each domains in FleS/FleR and characterized key residues in the FleS-HisKA, FleR-REC and FleR-AAA domains that are essential for the signaling. Next, we revealed that FleS/FleR regulates biofilm formation in a c-di-GMP and FleQ dependent manner. Lastly, we demonstrated that FleR can regulate flagellum biosynthesis independently without FleS, which explains the discrepant regulation of swimming motility by FleS and FleR.

Background: Human periodontal ligament stem cells (PDLSCs)-mediated regenerative periodontal therapy is a promising method for periodontitis. Besides, odontogenic stem cells are exposed to hypoxia in both physiological and pathological conditions. The behaviour of stem cells under hypoxia is worth exploring. So far, whether long noncoding RNAs (lncRNAs) can affect the proliferation and cementogenesis of PDLSCs, as well as their specific mechanism, remains to be elucidated. This study investigates lncRNA NEAT1 and its possible regulatory mechanisms of cementogenic differentiation of PDLSCs under hypoxia.Methods: CCK-8 assay, flow cytometry (FCM) and EdU assay was adopted to test the proliferation ability. Osteoblastic/cementogenic differentiation of PDLSCs under hypoxia was detected by western blot assay (WB), quantitative real-time polymerase chain reaction (RT-PCR), alkaline phosphatase (ALP) activity, alcian blue staining (ABS), alizarin red staining (ARS). The relationship between them was proved by dual-luciferase reporter assay and rescue experiments.Results: PDLSCs exhibited a more powerful cementogenic differentiation potential in hypoxic conditions. miR-214-5p may regulate SMAD4, while Lnc NEAT1 acts as a sponge, forming a ceRNA mechanism in which HIF-1α plays an important role. It may act as an enabler, jointly constitute the regulatory axis of Lnc NEAT1/ miR-214-5p/ SMAD4, and affect the cementogenic differentiation of PDLSCs under hypoxia.Conclusions: Our results showed that hypoxia could enhance the cementogenic differentiation potential of PDLSCs in vitro. In addition, we found that Lnc NEAT1 may be a potential target for improving the function of PDLSCs in regenerative periodontiology, and Lnc NEAT1/miR-214-5p/SMAD4 could regulate the cementogenic differentiation ability of PDLSCs under hypoxia.

Cripto is a small glycosylphosphatidylinositol-anchored signaling protein that can detach from the anchored membrane and stimulate proliferation, migration, differentiation, vascularization, and angiogenesis. In the present study, we demonstrated that Cripto positively affected proliferation and survival of mesenchymal stem cells (MSCs) without affecting multipotency. Cripto also increased expression of phosphorylated janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), 78 kDa glucose-regulated protein (GRP78), c-Myc, and cyclin D1. Notably, treatment with an anti-GRP78 antibody blocked these effects. In addition, pretreatment with STAT3 short interfering RNA (siRNA) inhibited the increase in p-JAK2, c-Myc, cyclin D1, and BCL3 levels caused by Cripto and attenuated the pro-survival action of Cripto on MSCs. We also found that incubation with Cripto protected MSCs from apoptosis caused by hypoxia or H2O2 exposure, and the level of caspase-3 decreased by the Cripto-induced expression of B-cell lymphoma 3-encoded protein (BCL3). These effects were sensitive to down-regulation of BCL3 expression by BCL3 siRNA. Finally, we showed that Cripto enhanced expression levels of vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), and hepatocyte growth factor (HGF). In summary, our results demonstrated that Cripto activated a novel biochemical cascade that potentiated MSC proliferation and survival. This cascade relied on phosphorylation of JAK2 and STAT3 and was regulated by GRP78. Our findings may facilitate clinical applications of MSCs, as these cells may benefit from positive effects of Cripto on their survival and biological properties.

Cynomolgus macaque has been used for the evaluation of the zoonotic potential of prion diseases, especially for classical-Bovine Spongiform Encephalopathy (classical-BSE) infectious agent. PrP amino acid sequence is considered to play a key role in the susceptibility to prion strains and only one amino acid change may alter this susceptibility. Macaque and human-PrP sequences have only nine amino acid differences, but the effect of these amino acid changes in the susceptibility to dissimilar prion strains is unknown. In this work, the transmissibility of a panel of different prions from several species was compared in transgenic mice expressing either macaque-PrPC (TgMac) or human-PrPC (Hu-Tg340). Similarities in the transmissibility of most prion strains were observed suggesting that macaque is an adequate model for the evaluation of human susceptibility to most of the prion strains tested. Interestingly, TgMac were more susceptible to classical-BSE strain infection than Hu-Tg340. This differential susceptibility to classical-BSE transmission should be taken into account for the interpretation of the results obtained in macaques. It could notably explain why the macaque model turned out to be so efficient (worst case model) until now to model human situation towards classical-BSE despite the limited number of animals inoculated in the laboratory experiments.

Semiochemicals play a pivotal role in the location, evaluation, and utilization of hosts by herbivorous insects. Mixtures of host plant-derived compounds are often required to elicit appropriate levels of response to olfactory stimuli. In multiple-choice bioassays, we characterized the response of adult Drosophila suzukii to foliage- and fruit-based synthetic compounds tested alone and in association with grape and tart cherry juices, and assessed whether synergistic interactions among olfactory stimuli are involved in the olfactory-driven behavior of D. suzukii. Our results established (1) significant attraction of females (but not males) to β-cyclocitral and isoamyl acetate when tested singly, (2) the presence of a synergistic interaction between β-cyclocitral and cherry juice only for females, and (3) the presence of a synergistic interaction between β-cyclocitral and isoamyl acetate but only in the case of males. Our findings increase our understanding of male and female D. suzukii olfactory responses to synthetic compounds and fruit juices as sources of attractants. Combinations of foliage- and fruit-based compounds may be needed to increase SWD attraction.

Objectives Ambient temperature impinges on energy metabolism in a body size dependent manner. This has implications for the housing temperature at which mice are best compared to humans. In 2013, we suggested that, for comparative studies, solitary mice are best housed at 23–25 °C, because this is 3–5 °C below the mouse thermoneutral zone and humans routinely live 3–5 °C below thermoneutrality, and because this generates a ratio of DEE to BMR of 1.6–1.9, mimicking the ratio found in free-living humans. Methods Recently, Fischer et al. (2017) challenged this estimate. By studying mice at 21 °C and at 30 °C (but notably not at 23–25 °C) they concluded that 30 °C is the optimal housing temperature. Here, we measured energy metabolism of C57BL/6 mice over a range of temperatures, between 21.4 °C and 30.2 °C. Results We observed a ratio of DEE to BMR of 1.7 at 27.6 °C and of 1.8 at 25.5 °C, suggesting that this is the best temperature range for housing C57BL/6 mice to mimic human thermal relations. We used a 24 min average to calculate the ratio, similar to that used in human studies, while the ratio calculated by Fisher et al. dependent on short, transient metabolic declines. Conclusion We concur with Fisher et al. and others that 21 °C is too cool, but we continue to suggest that 30 °C is too warm. We support this with other data. Finally, to mimic living environments of all humans, and not just those in controlled Western environments, mouse experimentation at various temperatures is likely required., Graphical abstract Image 1, Highlights • Translation from mice to man is best done at comparable metabolic conditions. • ‘Western’ humans metabolize at 1.7 to 1.8x BMR and are below thermoneutrality. • Thermoneutrality of solitary housed C57BL/6 laboratory mice commences above 28 °C. • 1.7x - 1.8x BMR for solitary housed C57BL/6 lab mice is between 25.5 °C - 27.6 °C. • Neither 21 °C nor 30 °C are suitable housing temperatures for translation.

Modified nano-graphene quantum dots (M-GQDs) are widely used in bioimaging, drug delivery, and chemical engineering. Because M-GQDs could induce reactive oxygen species and DNA damage, we hypothesized that M-GQDs modulate DNA methylation. To test this hypothesis, zebrafish were exposed to reduced, hydroxylated, or aminated GQDs (graphene quantum dots) at different concentrations for 7 days; global DNA methylation in liver, gill, and intestine was then studied. M-GQDs induced global DNA hypermethylation in various tissues in a dose-dependent manner. The global DNA methylation of reduced and aminated GQDs exposure showed a significant increase in intestines even at low concentrations (2 mg/L), suggesting that intestines are the main target for these two M-GQDs. The effects of global DNA methylation were evaluated 14 days after exposure had ceased. DNA methylation in the livers of exposure groups was significantly higher than in control zebrafish. Global DNA methylation increased in livers of zebrafish even after exposure to aminated GQDs (2 mg/L) had ceased, indicating a more complex mechanism of DNA methylation deregulation. The present results showed that chemical groups in the surface of GQDs are a critical factor for modulating DNA methylation.

Traumatic brain injuries introduce functional and structural circuit deficits that must be repaired for an organism to regain function. We developed an injury model in which Xenopus laevis tadpoles are given a penetrating stab wound that damages the optic tectal circuit and impairs visuomotor behavior. In tadpoles, as in other systems, injury induces neurogenesis. The newly generated neurons are thought to integrate into the existing circuit; however, whether they integrate via the same mechanisms that govern normal neuronal maturation during development is not understood. Development of the functional visuomotor circuit in Xenopus is driven by sensory activity. We hypothesized that enhanced visual experience would improve recovery from injury by facilitating integration of newly generated neurons into the tectal circuit. We labeled newly generated neurons in the injured tectum by green fluorescent protein expression and examined their circuit integration using electrophysiology and in vivo imaging. Providing animals with brief bouts of enhanced visual experience starting 24 h after injury increased synaptogenesis and circuit integration of new neurons and facilitated behavioral recovery. To investigate mechanisms of neuronal integration and behavioral recovery after injury, we interfered with N-methyl-d-aspartate (NMDA) receptor function. Ifenprodil, which blocks GluN2B-containing NMDA receptors, impaired dendritic arbor elaboration. GluN2B blockade inhibited functional integration of neurons generated in response to injury and prevented behavioral recovery. Furthermore, tectal GluN2B knockdown blocked the beneficial effects of enhanced visual experience on functional plasticity and behavioral recovery. We conclude that visual experience-mediated rehabilitation of the injured tectal circuit occurs by GluN2B-containing NMDA receptor-dependent integration of newly generated neurons.NEW & NOTEWORTHY Recovery from brain injury is difficult in most systems. The study of regenerative animal models that are capable of injury repair can provide insight into cellular and circuit mechanisms underlying repair. Using Xenopus tadpoles, we show enhanced sensory experience rehabilitates the injured visual circuit and that this experience-dependent recovery depends on N-methyl-d-aspartate receptor function. Understanding the mechanisms of rehabilitation in this system may facilitate recovery in brain regions and systems where repair is currently impossible.

Mesenchymal stem cells (MSCs) hold great potential to treat tissue damage based on their multipotent property, and are also considered as suitable cell resources to create tissue-engineered grafts for tendon repair. However, the clinical application of MSCs is still limited by the lack of efficient methods to induce tenogenic differentiation. In this study, by performing the experiments in transwell system, we found that paracrine factors from tenocytes could induce MSCs to undergo the tenogenic differentiation. We further verified that tenocytes could secrete exosomes and these tenocyte-derived exosomes efficiently initiated the tenogenic differentiation of MSCs. Finally, we revealed that the TGF-β existing in tenocyte-derived exosomes mediated the process, as the inhibition of TGF-β signaling abolished the effects of tenocyte-derived exosomes on MSCs. By investigating the effects of tenocytes on MSCs, we found that tenocytes-derived exosomes can induce tenogenic differentiation of MSCs in a TGF-β dependent manner. These studies provided critical information about the multipotency of MSCs and suggested potential strategies for clinical translation.

Store operated Ca(2+) entry (SOCE) is an ancient and ubiquitous Ca(2+) signaling pathway discovered decades ago, but the function of SOCE in human physiology is only now being revealed. The relevance of this pathway to striated muscle was solidified with the description of skeletal myopathies that result from mutations in STIM1 and Orai1, the two SOCE components. Here, we consider the evidence for STIM1 and SOCE in cardiac muscle and the sinoatrial node. We highlight recent studies revealing a role for STIM1 in cardiac growth in response to developmental and pathologic cues. We also review the role of STIM1 in the regulation of SOCE and Ca(2+) store refilling in a non-Orai dependent manner. Finally, we discuss the importance of this pathway in ventricular cardiomyocytes where SOCE contribute to developmental growth and in pacemaker cells where SOCE likely has a fundamental to generating the cardiac rhythm.

The boosting exploitation of graphene oxide (GO) increases exposure risk to human beings. However, as the first line defender, neutrophils’ mechanism of defensive behavior towards GO invasion remains unclear. Herein, we discover that neutrophils defensively degrade GO in a lateral dimension dependent manner. The micrometer-sized GO (mGO) induces NETosis by releasing neutrophil extracellular traps (NETs), while nanometer-sized GO (nGO) elicits neutrophil degranulation. The neutrophils’ defensive behavior is accompanied with generation of reactive oxygen species and activation of p-ERK and p-Akt kinases. We unveil that mGO-induced NETosis is NADPH oxidase (NOX)-independent, while nGO-triggered degranulation is NOX-dependent. Furthermore, myeloperoxidase (MPO) is identified to be a determinant mediator despite distinct neutrophil phenotypes in the biodegradation. Neutrophils release NETs comprising of MPO upon activated with mGO, while MPO is secreted via nGO induced-degranulation. Moreover, the binding energy between MPO and GO is calculated to be 69.8728 kJ·mol− 1, which indicates that electrostatic interactions mainly cause the spontaneous binding process in a spatial distance of 9.2 Å. Meanwhile, the central enzymatic biodegradation is found to occur at oxygenic active sites and defects on GO. Mass spectrometry analysis deciphers the degradation products are biocompatible molecules like flavonoids and polyphenols. Our study provides fundamental evidence and practical guidance for functional biomaterial development in sustainable nanotechnology, including but not limited to vaccine adjuvant and drug carrier.

SummaryThough gains of chromosome 12p13.31 are highly recurrent in hPSC, their impact on differentiation is poorly understood. We identify a reduction in differentiation capacity towards all three germ layers and a subpopulation of residual pluripotent cells that appear during hepatic specification. These cells form as a result of the overexpression of NANOG and GDF3, whereby NANOG as the primary driver delays activation of WNT signaling, partly as a result of a direct physical interaction with TCF7. Entry into the residual state is determined by cell cycle position at the onset of differentiation and is maintained by a feedback loop between NANOG and GDF3. These findings highlight the ability of genetically abnormal hPSC to escape correct differentiation and to form residual pluripotent cells, an important risk in the safe clinical translation of hPSC. Our results further refine the molecular mechanisms that underpin the exit from pluripotency and onset of differentiation.

BackgroundBACE inhibitors, which decrease BACE1 (β-secretase 1) cleavage of the amyloid precursor protein, are a potential treatment for Alzheimer’s disease. Clinical trials using BACE inhibitors have reported a lack of positive effect on patient symptoms and, in some cases, have led to increased adverse events, cognitive worsening and hippocampal atrophy. A potential drawback of this strategy is the effect of BACE inhibition on other BACE1 substrates such as Seizure-related gene 6 (Sez6) family proteins which are known to have a role in neuronal function.MethodsMice were treated with an in-diet BACE inhibitor for 4-8 weeks to achieve a clinically-relevant level of amyloid-β40 reduction in the brain. Mice underwent behavioural testing and postmortem analysis of dendritic spine number and morphology with Golgi-Cox staining. Sez6 family triple knockout mice were tested alongside wild-type mice to identify whether any effects of the treatment were due to altered cleavage of Sez6 family proteins.ResultsWild-type mice treated with BACE inhibitor displayed hyperactivity on the elevated open field, as indicated by greater distance travelled, but this effect was not observed in treated Sez6 triple knockout mice. BACE inhibitor treatment did not lead to significant changes in spatial or fear learning, reference memory, cognitive flexibility or anxiety in mice as assessed by the Morris water maze, context fear conditioning, or light-dark box tests. Chronic BACE inhibitor treatment reduced the density of mushroom-type spines in the somatosensory cortex, regardless of genotype, but did not affect steady-state dendritic spine density or morphology in the CA1 region of the hippocampus. ConclusionsChronic BACE inhibition for 1-2 months in mice led to increased locomotor output but did not alter memory or cognitive flexibility. While the mechanism underlying the treatment-induced hyperactivity is unknown, the absence of this response in Sez6 triple knockout mice indicates that blocking ectodomain shedding of Sez6 family proteins is a contributing factor. In contrast, the decrease in mature spine density in cortical neurons was not attributable to lack of shed Sez6 family protein ectodomains. Therefore, other BACE1 substrates are implicated in this effect and, potentially, in the cognitive decline in longer-term chronically treated patients.

Source-to-sink transport of sucrose mediated by sucrose transporters (SUCs) is one of the major determinants of plant growth. However, the role of AtSUC4, the only member of Group IV sucrose transporter in Arabidopsis, was undervalued in sink organ during seedling period. In our study, the primary root length of the atsuc4 mutants was significantly longer than that of the wild-type (WT) under exogenous 4% and 6% sucrose treatment. But this phenotype could not be imitated by external application of glucose or mannitol. It means that the atsuc4 mutants were insensitive to high sucrose stress compared with WT. Meanwhile, HPLC-MS/MS results showed that the root of atsuc4 mutants accumulated less sucrose and ABA and more IAA content compared with WT on 4% and 6% sucrose supplementation. Transcriptome analysis showed that many key genes involved in IAA and ABA signals were respective stimulated and repressed in the atsuc4 mutants, respectively. Taken together, we concluded that the deficiency of AtSUC4, not only reducing the transported and uptaked of sucrose, but also as a signal, may be collaborated with IAA and ABA to regulate root growth under high sucrose stress. This study confirmed the new function of AtSUC4, and provided an promising candidate gene for improving tolerance to high sucrose stress.

BackgroundIn spite of much progress in understanding the biology of cancer disease, advancement in technology for early diagnosis, the expanding array of anticancer drugs and treatment modalities, the global cancer burden is still significant and increasing. It is estimated that the new cases of cancer in the year 2040 will be 29.4 million per year globally. Sericin, an adhesive protein of silk cocoons has been shown to be a potential protein in various biomedical applications including cancer therapeutics. The present study evaluates the anticancer property of sericin from cocoons of Antheraea proylei J. (SAP) against human lung cancer (A549), cervical cancer (HeLa), and prostate cancer (PC3) cell lines. This is the first report of anti-cancer activity of the non-mulberry silkworm A. proylei J. Methods SAP was prepared from cocoons of A. proylei J. by the process of degumming method. The amino acid composition of SAP was determined by HPLC. Cytotoxicity activity was assessed by MTT assay and genotoxicity activity was assessed by comet assay. Cleavage of caspase and PARP proteins and phosphorylation of MAPK pathway members were analysed by Western blotting. Cell cycle analysis was done by flow cytometery.ResultsSAP causes cytotoxicity to A549, HeLa and PC3 cell lines with the IC50 values ranging from 3.4-3.9 µg/µl. SAP induces apoptosis in a dose-dependent manner through caspase-3 and p38 and ERK pathways in A549 and HeLa cells respectively whereas in PC3 cells, SAP induces apoptosis independent of caspase through p38 pathway. Moreover, in the case of A549 and HeLa cells SAP induces cell cycle arrest at S phase in a dose dependent manner whereas at G0 phase in the case PC3 cells.ConclusionSAP induces apoptosis in A549, HeLa, and PC3. The difference in the molecular mechanisms of apoptosis induced by SAP in A549 and HeLa and in PC3 may be due to the differences in the genotypes of the cancer cell lines. However, further investigation is warranted. The overall results of the present study envisage the possibility of using SAP as anti-tumorigenic agent.

Exercise and circadian biology are closely intertwined with physiology and metabolism, yet the functional interaction between circadian clocks and exercise capacity is only partially characterized. Here, we tested different clock mutant mouse models to examine the effect of the circadian clock and clock proteins, namely PERIODs and BMAL1, on exercise capacity. We found that daytime variance in endurance exercise capacity is circadian clock controlled. Unlike wild-type mice, which outperform in the late compared with the early part of their active phase, PERIODs- and BMAL1-null mice do not show daytime variance in exercise capacity. It appears that BMAL1 impairs and PERIODs enhance exercise capacity in a daytime-dependent manner. An analysis of liver and muscle glycogen stores as well as muscle lipid utilization suggested that these daytime effects mostly relate to liver glycogen levels and correspond to the animals' feeding behavior. Furthermore, given that exercise capacity responds to training, we tested the effect of training at different times of the day and found that training in the late compared with the early part of the active phase improves exercise performance. Overall, our findings suggest that clock proteins shape exercise capacity in a daytime-dependent manner through changes in liver glycogen levels, likely due to their effect on animals' feeding behavior.

Immune development is greatly influenced by vertically transferred cues. However, beyond B cells, little is known about the role of other subsets of the maternal immune system in regulating offspring immunity. We found that mice born from γδT cell-deficient (TCRδ-/-) dams display, early after birth, a pulmonary milieu selectively enriched in type-2 cytokines such as IL-33, IL-4, IL-5, and IL-13, and type-2-polarized immune cells, when compared to the progeny of γδT cell-sufficient dams. In addition, upon helminth infection, mice born from TCRδ-/- dams sustained an increased type-2 inflammatory response. Critically, this was independent of the genotype of the pups. The intestinal microbiota in the offspring of TCRδ-/- and TCRδ+/- dams harbored distinct bacterial communities acquired during birth and fostering, which were accompanied by changes in intestinal short-chain fatty acids (SCFA) profiles. Importantly, antibiotic treatment abrogated the differences in the pulmonary milieu, and exogenous SCFA supplementation suppressed first-breath- and infection-induced inflammation.

Simple Summary: On the basis of the extensive literature, two main strategies have been used to manipulate intestinal microbial composition and selectively stimulate the growth and activity of certain species, these being the administration of either prebiotics or food supplements containing living bacteria such as probiotics. Several animal studies have indicated that certain probiotics, including Lactobacilli and Bifidobacteria, can suppress body weight gain in rodents, while some probiotics strains have little effect or promote weight gain. The potential anti-obesity effect of probiotics seems to depend on the strains used and the underlying mechanisms, leading to their effects remaining not fully understood. It is in this context that this study was designed to investigate the potential of two probiotics strains, these being Bifidobacterium animalis subsp. lactis BB-12® and Lactobacillus plantarum 299v® in rabbits, whereby obesity and metabolic syndrome was first induced in a first experiment, and the animals were then used in a second experiment to test the hypothesis of probiotics effect on biochemical and morphometric parameters. The model of obesity induced by giving a “cafeteria” diet for 14 weeks in this trial demonstrated a change in the biochemical and morphometric parameters investigated in the ITELV2006 rabbit strain. This study revealed that B. animalis subsp. lactis BB-12 and L. plantarum 299v strains could exert beneficial effects in reducing the incidence of obesity and metabolic syndrome in the ITELV2006 rabbit strain. Abstract: This study aimed first to develop an experimental model of obesity and metabolic syndrome over 14 weeks using a diet called “cafeteria”, which is a high-fat diet, to evaluate its consequences on the biochemical and morphometric parameters in ITELV2006 strain rabbits. Second, the trial aimed to evaluate the effect of two strains of probiotics, these being Bifidobacterium animalis subsp. lactis BB-12® and Lactobacillus plantarum 299v®, on the obesity and MetS induced during the first experiment. Overall, the results of the “cafeteria” diet demonstrated significant changes in numerous biochemical and morphometric parameters, reproducing obesity and the main clinical manifestations of the metabolic syndrome in humans. The administration of the two probiotic strains demonstrated an impact on certain parameters of obesity and induced MetS. This study makes it possible to conclude that probiotics could be useful in the treatment of obesity and metabolic syndrome of rabbits, but in a dependent manner. Furthermore, this study evidenced the importance of selecting specific probiotic strains and dosages to achieve desirable results on rabbits or other species., Veterinary Institute Sciences, El-Khroub, Algeria, D01N01UN250120180004

The most successful anti-cancer drugs in clinical use interfere with cell cycle progression by inducing a mitotic arrest through activation of a conserved surveillance mechanism called the spindle assembly checkpoint (SAC). Cancer cells require much more glucose compared to normal cells to meet their energy needs due to reprogramming of their energy metabolism. Therefore, hyperglycemia may be favorable for cancer cells. Indeed, hyperglycemia is known to be closely associated with increased risk of developing several types of cancers as well as cancer related-mortality. However, the effect of hyperglycemic conditions on the mitotic arrest induced by anti-cancer drugs remains unknown. In this study, we investigated the effect of incubation in high glucose containing media on the duration of mitotic arrest induced by a SAC-activating anti-cancer drug, nocodazole, in budding yeast. We first showed that high glucose led to cell death in a dose and time dependent manner in yeast. Next, we examined the effect of sublethal concentrations of high glucose on nocodazole-induced mitotic arrest. Our data revealed that high glucose prolongs nocodazole induced mitotic arrest. Finally, we investigated whether the delay observed in exiting from nocodazole-induced mitotic arrest requires SAC activity. For this purpose, we induced a mitotic arrest that is independent of SAC activation and showed that the delay in exiting from nocodazole-induced mitotic arrest required SAC activity.

Copyright: © 2021 by the authors. Friedreich’s ataxia (FRDA) is a comparatively rare autosomal recessive neurological disorder primarily caused by the homozygous expansion of a GAA trinucleotide repeat in intron 1 of the FXN gene. The repeat expansion causes gene silencing that results in deficiency of the frataxin protein leading to mitochondrial dysfunction, oxidative stress and cell death. The GAA repeat tract in some cases may be impure with sequence variations called interruptions. It has previously been observed that large interruptions of the GAA repeat tract, determined by abnormal MboII digestion, are very rare. Here we have used triplet repeat primed PCR (TP PCR) assays to identify small interruptions at the 5′ and 3′ ends of the GAA repeat tract through alterations in the electropherogram trace signal. We found that contrary to large interruptions, small interruptions are more common, with 3′ interruptions being most frequent. Based on detection of interruptions by TP PCR assay, the patient cohort (n = 101) was stratified into four groups: 5′ interruption, 3′ interruption, both 5′ and 3′ interruptions or lacking interruption. Those patients with 3′ interruptions were associated with shorter GAA1 repeat tracts and later ages at disease onset. The age at disease onset was modelled by a group-specific exponential decay model. Based on this modelling, a 3′ interruption is predicted to delay disease onset by approximately 9 years relative to those lacking 5′ and 3′ interruptions. This highlights the key role of interruptions at the 3′ end of the GAA repeat tract in modulating the disease phenotype and its impact on prognosis for the patient. European Union Seventh Framework Programme (FP7/2007–2013) under grant agreement number 242193/EFACTS; National Brain Appeal—Small Acorns Fund; The Wellcome Centre for Human Neuroimaging is supported by core funding from the Wellcome (203147/Z/16/Z); Department of Health’s National Institute for Health Research Biomedical Research Centres funding scheme; CRN; North Thames, National Institute for Health Research (NIHR). Medical Research Council (MR/N028767/1).

Background: Glypican-3 (GPC3) mRNA was more frequently overexpressed in women and patients with invasive HCC. We explore possible molecular mechanisms of the effect of GPC3 on growth factor receptor-bound protein 10 (Grb10) and insulin-like growth factor 1 receptor (IGF-1R) interaction of tumor invasion in women. Methods: For in vitro experiments, GPC3 and pertinent mutants were transfected, and Western blotting (HEK293T cells), confocal microscopy (HeLa and PLC-PRF-5 cells), luciferase assays for AP-1 reporter activities (NIH3T3 and HuH-7 cells), gelatin zymography (PLC-PRF-5 cells) and cell culture in 3D collagen I gels (NIH3T3 and R- cells) were performed. For in vivo experiments, GPC3 and IGF-1R coexpression was evaluated in hepatocellular carcinoma clinical samples. Results: We found that interaction of IGF-1R with Grb10 was hindered by GPC3, and GPC3 causes IGF-1R colocalization with Grb10 to a lesser extent after IGF-1 stimulation; moreover, it promoted IGF-1-stimulated AP-1 activation and matrix metalloproteinase -2 and 9 (MMP-2 and MMP-9) secretion in vitro, which seemingly play a role in tumor invasion or recurrence. Further, gender differences existed among patients with hepatocellular carcinoma in terms of GPC3 and IGF-1R coexpression in vivo.Conclusions: We believe that a more intensive surveillance of GPC3 expression in female patients with hepatocellular carcinoma should contribute to the prediction of recurrence, and this may highlight new strategies for treating hepatocellular carcinoma in women.

Candida albicans as an opportunistic pathogen exploits the host immune system and causes a variety of life-threatening infections. The polymorphic nature of this fungus gives it tremendous advantage to breach mucosal barriers and cause a variety of oral and disseminated infections. Enterococcus faecalis, another opportunistic pathogen co-exists with C. albicans in several niches in the human body, including the oral cavity and gastrointestinal tract. However, interactions between E. faecalis and C. albicans on oral mucosal surfaces remain unknown. Here, for the first time, we comprehensively characterized the interactive profiles between laboratory and clinical isolates of C. albicans (SC5314 and BF1) and E. faecalis (OG1RF and 846) on an organotypic oral mucosal model. Our results demonstrated that the two species formed robust biofilms on the mucosal tissue surface with profound surface erosion and fungal invasion. Specifically, this effect was more pronounced in the laboratory isolates than in the clinical isolates. Notably, several genes of C. albicans involved in tissue adhesion, hyphal formation, fungal invasion, and biofilm formation were significantly upregulated in the presence of E. faecalis. This study highlights the strain-dependent cross-kingdom interactions between E. faecalis and C. albicans on oral mucosa, demonstrating the requisite to study more substrate-dependent polymicrobial interactions.

Numerous works have demonstrated that trypanosomatid motility is relevant for parasite replication and sensitivity. Nonetheless, although some findings indirectly suggest that motility also plays an important role during infection, this has not been extensively investigated. This work is aimed at partially filling this void for the case of Trypanosoma cruzi. After recording swimming T. cruzi trypomastigotes (CL Brener strain) and recovering their individual trajectories, we statistically analyzed parasite motility patterns. We did this with parasites that swim alone or above monolayer cultures of different cell lines. Our results indicate that T. cruzi trypomastigotes change their motility patterns when they are in the presence of mammalian cells, in a cell-line dependent manner. We further performed infection experiments in which each of the mammalian cell cultures were incubated for 2 h together with trypomastigotes, and measured the corresponding invasion efficiency. Not only this parameter varied from cell line to cell line, but it resulted to be positively correlated with the corresponding intensity of the motility pattern changes. Together, these results suggest that T. cruzi trypomastigotes are capable of sensing the presence of mammalian cells and of changing their motility patterns accordingly, and that this might increase their invasion efficiency.