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The evolutionarily conserved YAP/TAZ mechano-responsive transcription cofactors regulate development and tumorigenesis. Here, we show for the first time that human natural killer (NK) cells specifically express TAZ but not YAP, and TAZ serves to limit NK cytotoxicity. The TLR3 agonist, hiltonol, was able to trigger cytoplasmic sequestration of TAZ, which increased NK cytotoxicity. Mechanistically, a low dose of hiltonol enhanced the intracellular contractility of NK cells accompanied by an increase in active RhoA and myosin light chain phosphorylation, conceivably through an increase in ERK1/2 activation-dependent ROS production. Importantly, we showed that the dissociation of LATS1 from actin upon activation of contractility, is required to sequester TAZ in the cytoplasm. Functionally, hiltonol also reduced the NK cell surface inhibitory receptors, KIR3DL1 through c-Myc inhibition and PD-1 through enhanced contractility. Direct inhibition of c-Myc promoted NK cytotoxicity against K562 lymphoblast. We corroborated our findings by coculturing hiltonol-pretreated NK cells with breast and lung cancer cells, and demonstrated increased NK-mediated cancer killing, which conceivably occurred via sequestration of TAZ to the cytoplasm which facilitated NK cytotoxicity. Our findings pave the way forex vivorejuvenation of NK cells forin vivoimmunotherapies, which could involve a cocktail of hiltonol and c-Myc inhibitor.

Accurate and early estimation of the thermal stability factor of a memory array is essential for the development of modern non-volatile magnetic storage devices. We propose an evaluation method for effective array-level thermal stability factors ( $\Delta _{\mathrm {eff}}$ ) via magnetic characterization of patterned magnetic tunnel junction (MTJ) arrays. These patterned arrays do not require traditional electrical contacts and circuitry that are usually needed for $\Delta _{\mathrm {eff}}$ evaluation. Using vibrating sample magnetometer (VSM) at elevated temperatures and under applied fields, we demonstrate that these MTJ arrays exhibit a moment change trend that fits the Neel–Brown model and are able to extract the respective $\Delta _{\mathrm {eff}}$ . The domain-wall-mediated reversal model is used to obtain zero-field $\Delta _{\mathrm {eff}}$ . Using this method, we have optimized MTJ film stacks based on fast array-level feedback and validated high data retention of $\Delta _{\mathrm {eff}} \sim 46$ at 260 °C from fully integrated MTJ bit arrays.

Circulating Ly6Chi monocytes often undergo cellular death upon exhaustion of their antibacterial effector functions, which limits their capacity for subsequent macrophage differentiation. This shrouds the understanding on how the host replaces the tissue-resident macrophage niche effectively during bacterial invasion to avert infection morbidity. Here, we show that proliferating transitional premonocytes (TpMos), an immediate precursor of mature Ly6Chi monocytes (MatMos), were mobilized into the periphery in response to acute bacterial infection and sepsis. TpMos were less susceptible to apoptosis and served as the main source of macrophage replenishment when MatMos were vulnerable toward bacteria-induced cellular death. Furthermore, TpMo and its derived macrophages contributed to host defense by balancing the proinflammatory cytokine response of MatMos. Consequently, adoptive transfer of TpMos improved the survival outcome of lethal sepsis. Our findings hence highlight a protective role for TpMos during bacterial infections and their contribution toward monocyte-derived macrophage heterogeneity in distinct disease outcomes. Agency for Science, Technology and Research (A*STAR) National Research Foundation (NRF) Published version This research was funded by Singapore Immunology Network (SIgN) core funding and A*STAR, Singapore. F.G. is an EMBO YIP awardee and is supported by SIgN core funding and Singapore National Research Foundation Investigatorship (NRF2016NRF-NRF1001-02). L.G.N. is supported by SIgN core funding. S.Z.C. is supported by the Ministry of Health, Singapore’s National Medical Research Council under its Open Fund-Young Individual Research Grant (OFYIRG17may036), and A*STAR Career Development Award (192D8043). Y.C.T. is a graduate scholar supported by the Ministry of Education, Singapore. M.Y.C. is a graduate scholar supported by the SIgN-NTU-NUS scholarship, Singapore. SIgN Flow Cytometry facility is supported by National Research Foundation (NRF) Singapore under Shared Infrastructure Support (SIS) (NRF2017_SISFP09).

Macrophages (Mϕ) have been reported to downmodulate the cytotoxicity of natural killer (NK) cell against solid tumor cells. However, the collaborative role between NK cells and Mϕ remains underappreciated, especially in hematological cancers, such as chronic myeloid leukemia (CML). We observed a higher ratio of innate immune cells (Mϕ and NK) to adaptive immune cells (T and B cells) in CML bone marrow aspirates, prompting us to investigate the roles of NK and Mϕ in CML. Using coculture models simulating the tumor inflammatory environment, we observed that Mϕ protects CML from NK attack only when CML was itself mycoplasma‐infected and under chronic infection–inflammation condition. We found that the Mϕ‐protective effect on CML was associated with the maintenance of CD16 level on the NK cell membrane. Although the NK membrane CD16 (mCD16) was actively shed in Mϕ + NK + CML trioculture, the NK mCD16 level was maintained, and this was independent of the modulation of sheddase by tissue inhibitor of metalloproteinase 1 or inhibitory cytokine transforming growth factor beta. Instead, we found that this process of NK mCD16 maintenance was conferred by Mϕ in a contact‐dependent manner. We propose a new perspective on anti‐CML strategy through abrogating Mϕ‐mediated retention of NK surface CD16., The collaborative role between natural killer (NK) cells and macrophages (Mϕ) remains unappreciated in hematological tumors. Here, we found that mycoplasma‐infected chronic myeloid leukemia (CML) cells were protected by Mϕ from NK cytotoxicity and this protective effect was associated with Mϕ‐mediated maintenance of NK membrane CD16 level in a contact‐dependent manner. Further studies would shed light into potential therapeutics for CML via modulation of interactions between innate immune cells, NK and Mϕ.

Natural killer (NK) cells are innate immune cells which play a key role in shaping the immune response against cancer. Initially hailed for their potential to recognise and eliminate tumour cells, their application has been greatly hindered by the immunosuppressive tumour microenvironment (TME) which suppresses NK functions (e.g., cytotoxicity). This dysfunctional state that is accompanied by phenotypic changes such as upregulation of inhibitory receptors and downregulation of activating receptors, forms the basis of what many researchers have referred to as ‘exhausted’ NK cells. However, there is no consensus on whether these phenotypes are sufficient to define an exhausted state of the NK cell. While recent advances in checkpoint inhibition appear to show promise in early-stage pre-clinical studies, much remains to be fully explored and understood in the context of the TME. The TME is where the NK cells are subjected to interaction with various cell types and soluble factors, which could exert an inhibitory effect on NK cytotoxicity. In this review, we provide an overview of the general markers of NK cell exhaustion viz, the surface activating and inhibitory receptors. We also highlight the potential role of T-box transcription factors in characterising such a dysfunctional state and discuss the often-overlooked mechanism of cell cytoskeletal dynamics in regulating NK cell function. These aspects may further contribute to NK exhaustion or NK revival in cancer and may open new avenues to explore cancer treatment strategies.

Simple Summary The dysregulation of UPS exacerbates the tumor microenvironment and drives malignant transformation. As the largest family of E3 ligases, the SCFF-boxes promotes BRCA progression. FBXL8 was recently identified to be a novel SCF E3 ligase that potently promotes BRCA. Here, we profiled the transcriptome of BRCA patient tissues by global NGS RNA-Seq and TCGA database analyses. A signature of four SCFF-box E3 ligases (FBXL8, FBXO43, FBXO15, CCNF) was found to be pivotal for BRCA advancement. Knockdown of FBXL8 and FBXO43 reduced cancer cell viability and proliferation, suggesting their pro-tumorigenic roles. However, the overexpression of CCNF inhibited cancer cell progression, indicating its anti-tumorigenic role. FBXL8 and FZR1 pulled down CCNF, and double knockdown of FBXL8 and FZR1 caused CCNF accumulation. Additionally, CCNF partnered with a pro-tumorigenic factor, RRM2, and overexpression of CCNF reduced RRM2. Our findings suggest a potential for drugging CCNF in co-modulatory partnership with FBXL8 and FZR1, for anti-BRCA therapy. Abstract Breast cancer (BRCA) malignancy causes major fatalities amongst women worldwide. SCF (Skp1-cullin-F-box proteins) E3 ubiquitin ligases are the most well-known members of the ubiquitination–proteasome system (UPS), which promotes cancer initiation and progression. Recently, we demonstrated that FBXL8, a novel F-box protein (SCFF-boxes) of SCF E3 ligase, accelerates BRCA advancement and metastasis. Since SCFF-boxes is a key component of E3 ligases, we hypothesized that other SCFF-boxes besides FBXL8 probably collaborate in regulating breast carcinogenesis. In this study, we retrospectively profiled the transcriptome of BRCA tissues and found a notable upregulation of four SCFF-box E3 ligases (FBXL8, FBXO43, FBXO15, and CCNF) in the carcinoma tissues. Similar to FBXL8, the knockdown of FBXO43 reduced cancer cell viability and proliferation, suggesting its pro-tumorigenic role. The overexpression of CCNF inhibited cancer cell progression, indicating its anti-tumorigenic role. Unexpectedly, CCNF protein was markedly downregulated in BRCA tissues, although its mRNA level was high. We showed that both E3 ligases, FBXL8 and FZR1, pulled down CCNF. Double knockdown of FBXL8 and FZR1 caused CCNF accumulation. On the other hand, CCNF itself pulled down a tumorigenic factor, RRM2, and CCNF overexpression reduced RRM2. Altogether, we propose a signature network of E3 ligases that collaboratively modulates CCNF anti-cancer activity. There is potential to target BRCA through modulation of the partnership axes of (i) CCNF-FBXL8, (ii) CCNF-FZR1, and (iii) CCNF-RRM2, particularly, via CCNF overexpression and activation and FBXL8/FZR1 suppression.

The initiation and progression of breast cancer (BRCA) is associated with inflammation and immune-overactivation, which is critically modulated by the E3 ubiquitin ligase. However, the underlying mechanisms and key factors involved in BRCA formation and disease advancement remains under-explored. By retrospective studies of BRCA patient tissues, and gene knockdown and gain/loss-of-function studies, we uncovered a novel E3 ligase, FBXL8, in BRCA. A signature expression profile of F-box factors that specifically target and degrade proteins involved in cell death/survival, was identified. FBXL8 emerged as a prominent member of the F-box factors. Ex vivo analysis of 1349 matched BRCA tissues indicated that FBXL8 promotes cell survival and tumorigenesis, and its level escalates with BRCA progression. Knockdown of FBXL8 caused: (i) intrinsic apoptosis, (ii) inhibition of cell migration and invasion, (iii) accumulation of two tumor-suppressors, CCND2 and IRF5, and (iv) downregulation of cancer-promoting cytokines/chemokines, all of which curtailed the tumor microenvironment and displayed potential to suppress cancer progression. Co-IP study suggests that two tumor-suppressors, CCND2 and IRF5 are part of the immune-complex of FBXL8. The protein levels of CCND2 and IRF5 inversely correlated with FBXL8 expression, implying that FBXL8 E3 ligase was associated with the degradation of CCND2 and IRF5. Altogether, we propose the exploitation of the ubiquitin signaling axis of FBXL8-CCND2-IRF5 for anti-cancer strategies and potential therapeutics.

The initiation and progression of breast cancer (BRCA) is associated with inflammation and immune-overactivation. The E3 ubiquitin ligase is known to subtly balance immune-overactivation and pro-tumorigenesis. Here, by global transcriptional profiling of BRCA patient tissues, we identified a signature expression profile of F-box factors, of which FBXL8 emerged as a novel key component of E3 ligase. Our ex vivo studies (n=1349) indicate that FBXL8 promotes tumorigenesis and its level escalates with BRCA advancement. Knockdown of FBXL8 caused: (i) accumulation of two tumor-suppressors,CCND2 and IRF5, suggesting their collaborative regulation of BRCA status, (ii) intrinsic apoptosis in BRCA, (iii) inhibition of cell migration and invasion and (iv) downregulation of cancer-promoting cytokines/ chemokines; all of these effects curtailed the tumor microenvironment and suppressed cancer progression. Our findings highlight the translational impact of exploiting FBXL8 and its interaction partners (CCND2 and IRF5) for developing anti-cancer strategies and potential therapeutics to limit BRCA progression.

Cells and soluble mediators of the innate and adaptive immune systems are fundamental components of the tumor microenvironment. Nuclear factors, e.g. transcription factors (TFs) and oncoproteins/cancer suppressors, play important roles in controlling cytokine functions leading to the development, maintenance and metastasis of cancers. Studies focusing on the regulators of the pro-tumorigenic microenvironment are particularly pertinent to early diagnosis and potential development of targeted cancer therapeutics. This review is motivated by new insights into the molecular dynamics of ubiquitination and SUMOylation, which post-translationally modify tumor suppressor TFs, leading to initiation and progression of various cancers like prostate, colorectal, liver and breast cancers. These modification pathways are differentially modulated under various stimuli or stresses in order to sustain the oncogenic potentials. We deliberate on the vicious cycle of infection and chronic inflammation-driven processes of ubiquitination and SUMOylation, resulting in the imbalance in cytokine profiles in the pro-tumorigenic microenvironment.

Novel properties of two-dimensional (2D) materials have drawn extensive attention in past decade. Magnetic property is one of the research focuses since it can add extra spin freedom for the manipulation of 2D material properties. The magnetism in 2D materials can be originated from defects (i.e. vacancies, zigzag defects). In this work, MoS2 nano particles were annealed at various temperatures (from 600 till 1000 °C) in an argon environment to explore the post-annealing effect on the magnetic properties. The results indicate that annealing may induce tunable ferromagnetism associated with the annealing temperature. The ferromagnetic signal is enhanced with the increase in annealing temperature which may attribute to the defects pertaining to the sulfur vacancies and edge terminated structure induced by annealing.

The ubiquitin-proteasome system (UPS) modulates carcinogenesis through ubiquitination of cancer-related target proteins, leading to their degradation in the proteasome. This may deactivate tumor suppressors or activate tumor promoters- either way causing homeostatic imbalance. As major components of the UPS, the E2 and E3 enzymes are recognized as pivotal determinants of substrate recognition and ubiquitination. Identification of E2-E3 pairing selectivity is particularly pertinent to early diagnosis and potential development of targeted cancer therapeutics. This review is motivated by recent findings and new insights into the molecular dynamics of ubiquitination triggered by specific E2-E3 pairing, leading to cancer initiation and progression if cancer suppressors are degraded or cancer suppression (if cancer promoters are degraded), respectively. We provide an overview of strategies employed in screening for E2-E3 interactions based on up-to-date studies focusing on the E2-E3 interface motifs. Of considerable recent interest is how E2 and E3 might switch their functional partnerships via UBE2O, which suggests an emerging significance on how UBE2O might influence E2-E3 pairing. Thus, a reflection on the role of UBE2O is included. Finally, we deliberate on the rational and cautious development of anti-cancer cocktail drugs which specifically target E2-E3 interacting residues for precision in cancer-killing with minimal side-effects. To this end, a list of potential future research is proposed.

In this research, the potential of transition metal doped 2D WSe2 as a 2D dilute magnetic semiconductor has been explored experimentally. Single crystal WSe2 was doped with Co with various concentrations by a physical implantation method. Raman and X-ray diffraction spectra indicate the effective doping of Co in the substitutional sites. Secondary ion mass spectrometry results demonstrate the Gaussian distribution of Co in WSe2. Magnetic measurements show that both un-doped WSe2 and WSe2 doped with Co exhibit weak ferromagnetism at room temperature. The magnetization is increased with Co doping compared to un-doped WSe2 and it tends to be enhanced with increasing doping concentration.

The effects of Mn doping on the ferromagnetic properties of the dilute magnetic semiconductor Zn 1−x Mn x O nanorods (NR’s) having the nominal composit-ions x = 0, 0.01, 0.03, 0.04 and 0.05 grown by a low temperature hydrothermal method are studied. Energy dispersive X-ray (EDX) is used to determine the actual amounts of the elements in each NR’s. X-ray diffraction, scanning electron microscopy, photoluminescence and vibrating sample magnetometer measurements are used to observe the effects of the Mn substitution on the properties of the doped ZnO and to relate the changes in the properties to changes in the defect content. It is observed that the saturation magnetization of the Mn ions in the wurtzite structure varies from 0.0210 µ B /Mn 2+ to 0.0234 µ B /Mn 2+ reaching a high of 0.0251 µ B /Mn 2+ as the Mn concentrations is varied from 0.9 to 7.36 atomic%. It is argued that the changes in the saturation magnetization are due to the competition between the direct Mn-Mn exchange interaction and the indirect Mn-O-Mn exchange interaction in the doped Mn ZnO NP’s.

Room temperature ferromagnetism has been observed in the Cu doped ZnO films deposited under an oxygen partial pressure of 10−3 and 10−5 torr on Pt (200 nm)/Ti (45 nm)/Si (001) substrates using pulsed laser deposition. Due to the deposition at relatively high temperature (873 K), Cu and Ti atoms diffuse to the surface and interface, which significantly affects the magnetic properties. Depth sensitive polarized neutron reflectometry method provides the details of the composition and magnetization profiles and shows that an accumulation of Cu on the surface leads to an increase in the magnetization near the surface. Our results reveal that the presence of the copper at Zn sites induces ferromagnetism at room temperature, confirming intrinsic ferromagnetism.

Recent clinical and therapeutic success with RAF and MEK1/2 inhibitors has revolutionized the existing treatment schemes for previously incurable cancers like melanomas. However, the overall therapeutic efficacies are still largely compromised by the dose-limiting side effects and emerging drug resistance mechanisms. Accumulating evidence has revealed the intricate nature of the RAS-RAF-MEK1/2-ERK1/2 pathway, such as activation mechanisms, kinase–substrate relationships, crosstalk with parallel signaling pathways, feedback regulations, and intimate interplay with immune responses. Limited strategies are currently available to exploit the benefits of combining RAF-MEK1/2-ERK1/2 pathway inhibitors with other targeted therapies or immunotherapies. Here, we compiled the kinase–substrate relationships and analyzed the intricate signaling networks of the renowned pathway, providing an integrated and simplified visualization, to reveal the potentials of RAS-RAF-MEK1/2-ERK1/2-based combination therapies.

The role of ribosomal protein S6 (rpS6) phosphorylation in mRNA translation remains poorly understood. Here, we reveal a potential role in modulating the translation rate of chemokine (C-X-C motif) ligand 8 (CXCL8 or Interleukin 8, IL8). We observed that more CXCL8 protein was being secreted from less CXCL8 mRNA in primary macrophages and macrophage-like HL-60 cells relative to other cell types. This correlated with an increase in CXCL8 polyribosome association, suggesting an increase in the rate of CXCL8 translation in macrophages. The cell type-specific expression levels were replicated by a CXCL8- UTR-reporter (Nanoluc reporter flanked by the 5’ and 3’ UTR of CXCL8). Mutations of the CXCL8-UTR-reporter revealed that cell type-specific expression required: 1) a 3’ UTR of at least three hundred bases; and 2) an AU base content that exceeds fifty percent in the first hundred bases of the 3’ UTR immediately after the stop codon, which we dub AU-rich proximal UTR sequences (APS). The 5’ UTR of CXCL8 enhanced expression at the protein level and conferred cell type-specific expression when paired with a 3’ UTR. A search for other APS-positive mRNAs uncovered TNF alpha induced protein 6 (TNFAIP6), another mRNA that was translationally upregulated in macrophages. The elevated translation of APS-positive mRNAs in macrophages coincided with elevated rpS6 S235/236 phosphorylation. Both were attenuated by the ERK1/2 signaling inhibitors, U0126 and AZD6244. In A549 cells, rpS6 S235/236 phosphorylation was induced by TAK1, Akt or PKA signaling. This enhanced the translation of the CXCL8-UTR-reporters. Thus, we propose that the induction of rpS6 S235/236 phosphorylation enhances the translation of mRNAs that contain APS motifs, such as CXCL8 and TNFAIP6. This may contribute to the role of macrophages as the primary producer of CXCL8, a cytokine that is essential for immune cell recruitment and activation., Author summary Ribosomal protein S6 (rpS6) is a component of the cell translation system. This system produces proteins based on the instructions found on messenger ribonucleic acids or mRNAs. The activity of rpS6 is modified via the attachment of phosphate groups. This rpS6 “phosphorylation” regulates cell size, cell proliferation, glucose homeostasis and cancer defense. However, the molecular mechanism remains poorly understood. Here, we propose that rpS6 phosphorylation may affect the rate of translation of certain mRNAs. These mRNAs contain a high frequency of adenine and uracil bases in the first hundred or so bases of their 3’ UTR sequences, which we dub AU-rich proximal UTR sequences (APS). APS-modulated mRNAs include the immune signaling proteins, CXCL8 and TNFAIP6. This novel mechanism allows macrophages to secrete sufficient CXCL8 protein to attract and activate other immune cells such as neutrophils during injury or disease. It may also be aberrantly upregulated in a subset of tumors, leading to the overexpression of APS-positive genes such as CXCL8. Indeed, both rpS6 phosphorylation and CXCL8 expression are consistently found, by separate studies, to be tumor-promoting and upregulated in cancers. Modulating these APS-positive mRNAs may be a novel strategy to treat diseases.

// Wai Hoe Lau 1 , Xiphias Ge Zhu 1 , Shamaine Wei Ting Ho 1 , Shu Chun Chang 1, 2 and Jeak Ling Ding 1 1 Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore 117543, Singapore 2 Taipei Medical University, College for Medical Science and Technology, Taipei 110, Taiwan Correspondence to: Jeak Ling Ding, email: dbsdjl@nus.edu.sg Keywords: lung cancer cells, polyI:C-TLR3 suppression of survival and metastasis, anti-IL6 antibody, JAK2/STAT3 antagonists, cytokines and caspase 3/7 apoptosis Received: November 30, 2016 Accepted: February 07, 2017 Published: March 02, 2017 ABSTRACT Activation of TLR3 stimulates cancer cell apoptosis and triggers secretion of inflammatory cytokines. PolyI:C, a TLR3 agonist, activates immune cells and regresses metastatic lung cancer in vivo . Although polyI:C reportedly kills lung carcinomas, the mechanism remains elusive. Here, we demonstrated that polyI:C suppressed the proliferation and survival of metastatic (NCI-H358 and NCI-H292) and non-metastatic (A549) lung cancer cells. Notably, A549, NCI-H292 and NCI-H358 which are inducible by polyI:C, expressed low-to-medium level of TLR3 protein, and were susceptible to polyI:C treatment. By contrast, NCI-H1299, which endogenously expresses high level of TLR3 protein, was insensitive to polyI:C. We showed that polyI:C stimulated pro-inflammatory cytokines associated with survival and metastasis in a cell type-specific manner. While A549 and NCI-H292 released high levels of IL6, IL8 and GRO, the NCI-H358 cells endogenously secretes abundant levels of these cytokines, and was not further induced by polyI:C. Thus, NCI-H358 was resistant to the inhibition of cytokine-dependent metastasis. NCI-H1299, which was unresponsive to polyI:C, did not produce any of the pro-inflammatory cytokines. Treatment of A549 with a combination of polyI:C and anti-IL6 antibody significantly decreased IL6 production, and enhanced polyI:C-mediated killing and suppression of oncogenicity and metastasis. While polyI:C stimulated the phosphorylation of STAT3 and JAK2, blockade of these proteins enhanced polyI:C-mediated suppression of survival and metastasis. Taken together, polyI:C alone provoked apoptosis of lung cancer cells that express low-to-medium levels of functional TLR3 protein. The combinatorial treatment with polyI:C and anti-IL6 enhanced polyI:C-mediated anticancer activities through IL6/JAK2/STAT3 signalling, and apoptosis via TLR3-mediated caspase 3/8 pathway.

Al doped ZnO nanorods (NR's) having Al concentration up to 10 mol% were grown by the hydrothermal method. XRD measurements showed that the Al substituted ZnO NR's maintained the hexagonal wurtzite structure for all levels of Al substitution. EDX measurements of the ZnO:Al NR's indicated that the Al substitution created additional Zn vacancies in the wurtzite structure which is reflected in the enhanced photoluminescence emission in the visible light spectra between 450 and 550 nm of the more heavily doped ZnO:Al NR's. SEM images of the heavier doped ZnO:Al nanorods showed nano nodules being formed on the surface of the hexagonal shaped NR's. The saturation magnetizations of the ZnO:Al NR's as measured by a SQUID magnetometer increased to 10.66×10 −4 emu/g as more Al was substituted in. The hysteresis loops for the ZnO:Al NR's began to exhibit novel effects, such as horizontal shift (exchange bias field 0.0382 kOe for the 9 mol% NR) and butterfly shapes.

NSCLC (non-small cell lung cancer) is a leading cause of cancer-related deaths worldwide. Clinical trials showed that Hiltonol, a stable dsRNA representing an advanced form of polyI:C (polyinosinic-polycytidilic acid), is an adjuvant cancer-immunomodulator. However, its mechanisms of action and effect on lung cancer have not been explored pre-clinically. Here, we examined, for the first time, how a novel Hiltonol cocktail kills NSCLC cells. By retrospective analysis of NSCLC patient tissues obtained from the tumor biobank, pre-clinical studies with Hiltonol alone or Hiltonol+++ cocktail [Hiltonol+anti-IL6+AG490 (JAK2 inhibitor)+Stattic (STAT3 inhibitor)], cytokine analysis, gene knockdown and gain/loss-of-function studies, we uncovered the mechanisms of action of Hiltonol+++. We demonstrated that Hiltonol+++ kills the cancer cells and suppresses the metastatic potential of NSCLC through: (i) upregulation of pro-apoptotic Caspase-9 and Caspase-3, (ii) induction of cytosolic cytochrome c, (iii) modulation of pro-inflammatory cytokines (GRO, MCP-1, IL-8, and IL-6) and anticancer IL-24 in NSCLC subtypes, and (iv) upregulation of tumor suppressors, PKR (protein kinase R) and OAS (2′5′ oligoadenylate synthetase). In silico analysis showed that Lys296 of PKR and Lys66 of OAS interact with Hiltonol. These Lys residues are purportedly involved in the catalytic/signaling activity of the tumor suppressors. Furthermore, knockdown of PKR/OAS abrogated the anticancer action of Hiltonol, provoking survival of cancer cells. Ex vivo analysis of NSCLC patient tissues corroborated that loss of PKR and OAS is associated with cancer advancement. Altogether, our findings unraveled the significance of studying tumor biobank tissues, which suggests PKR and OAS as precision oncological suppressor candidates to be targeted by this novel Hiltonol+++ cocktail which represents a prospective drug for development into a potent and tailored therapy for NSCLC subtypes.

The name of the first author in the article by Tulisas et al. [(2009), Acta Cryst. F65, 533–535] is corrected.

The transcription factor, T‐bet, regulates type 1 inflammatory responses against a range of infections. Here, we demonstrate a previously unaddressed role of T‐bet, to influenza virus and bacterial superinfection. Interestingly, we found that T‐bet deficiency did not adversely affect the efficacy of viral clearance or recovery compared to wild‐type hosts. Instead, increased infiltration of neutrophils and production of Th17 cytokines (IL‐17 and IL‐22), in lungs of influenza virus‐infected T‐bet−/− mice, were correlated with survival advantage against subsequent infection by Streptococcus pneumoniae. Neutralization of IL‐17, but not IL‐22, in T‐bet−/− mice increased pulmonary bacterial load, concomitant with decreased neutrophil infiltration and reduced survival of T‐bet−/− mice. IL‐17 production by CD8+, CD4+ and γδ T cell types was identified to contribute to this protection against bacterial superinfection. We further showed that neutrophil depletion in T‐bet−/− lungs increased pulmonary bacterial burden. These results thus indicate that despite the loss of T‐bet, immune defences required for influenza viral clearance are fully functional, which in turn enhances protective type 17 immune responses against lethal bacterial superinfections.

Monocytes are a subset of cells that are categorized together with dendritic cells (DCs) and macrophages in the mononuclear phagocyte system (MPS). Despite sharing several phenotypic and functional characteristics with MPS cells, monocytes are unique cells with the ability to function as both precursor and effector cells in their own right. Before the development of hematopoietic stem cells (HSCs) in utero, monocytes are derived from erythro-myeloid precursors (EMPs) in the fetal liver that are important for populating the majority of tissue resident macrophages. After birth, monocytes arise from bone marrow (BM)-derived HSCs and are released into the circulation upon their maturation, where they survey peripheral tissues and maintain endothelial integrity. Upon sensing of microbial breaches or inflammatory stimuli, monocytes migrate into tissues where their plasticity allows them to differentiate into cells that resemble macrophages or DCs according to the environmental niche. Alternatively, they may also migrate into tissues in the absence of inflammation and remain in an undifferentiated state where they perform homeostatic roles. As monocytes are typically on the move, the availability of intravital imaging approaches has provided further insights into their trafficking patterns in distinct tissue compartments. In this review, we outline the importance of understanding their functional behavior in the context of tissue compartments, and how these studies may contribute towards improved vaccine and future therapeutic strategies. NMRC (Natl Medical Research Council, S’pore) Published version

Properties of particles whose dimensions are of the nanoscale can be quite different from those of bigger dimension particles. We investigate here the effects of doping Sb 5+ ions into ZnO nanorods (NR's) on the formation of magnetic moments. The pure ZnO and Sb-doped ZnO NR's were grown by the hydrothermal process. The fabricated nanorods were then studied using X-ray diffraction (XRD), ultraviolet–visible light spectroscopy (UV–vis), Scanning Electron Microscopy (SEM), Photoluminescence (PL) Spectrometer and Vibrating Sample Magnetometer (VSM) spectrometry. Substitution of Sb into the ZnO leads to the formation of a defect (Sb Zn +2V Zn ) complex state which can capture the oxygen p- orbital electrons of the oxygen atoms and form a spin polarized state. The PL was used to monitor the defect formation in the ZnO NR's when the Sb is doped in. As more Sb was doped in, the visible blue portion of the PL spectrum decreased while the visible red portion increased. Magnetization measurements (VSM) showed that the pure ZnO NR's possessed a magnetic moment which initially increased as Sb is doped in but decreased as more Sb was doped in. We obtain a value of 1.015 memu/g for the saturation magnetization of the pure ZnO nanorod.

Microbial pathogens are responsible for most of the rapidly-spreading deadly infectious diseases against humans. Thus, there is an urgent need for efficient and rapid detection methods for infectious microorganisms. The detection methods should not only be targeted and specific, but they have to be encompassing of potential changes of the pathogen as it evolves and mutates quickly during an epidemic or pandemic. The existing diagnostics such as the antibody-based ELISA immunoassay and PCR methods are too selective and narrowly focused; they are insufficient to capture newly evolved mutant strains of the pathogen. Here, we introduce a fresh perspective on some new technologies, including aptamers and next generation sequencing for pathogen detection. These technologies are not in their infancy; they are reasonably mature and ready, and they hold great promise for unparalleled applications in pathogen detection.

SARM (Sterile alpha and armadillo motif-containing protein) is the recently identified TIR domain-containing cytosolic protein. Classified as a member of the TLR adaptor family, the multiple locations and functions of SARM (sometimes playing opposing roles), provoke an enigma on its biology. Although originally assumed to be a member of the TLR adaptor family (functioning as a negative regulator of TLR signaling pathway), latest findings indicate that SARM regulates signaling differently from other TLR adaptor proteins. Recent studies have highlighted the significant functional role of SARM in mediating apoptosis and antiviral innate immune response. In this review, we provide an update on the evolutionary conservation, spatial distribution, and regulated expression of SARM to highlight its diverse functional roles. The review will summarize findings on the known interacting partners of SARM and provide analogy on how they add new dimensions to the current understanding on the multifaceted roles of SARM in antiviral activities and apoptotic functions. In addition, we provide a future perspective on the roles of SARM in differentiation and development, with substantial emphasis on the molecular insights to its mechanisms of action.

SrRuO3 films were prepared using pulsed laser deposition on different substrates (z-cut Quartz, (001) SrTiO3 and (001) LaAlO3) in this work. We have observed greatly enhanced perpendicular coercivity over 2 T in nanocrystalline SrRuO3 thin films with a thickness of 60 nm deposited at 600 °C on quartz substrates compared to epitaxial thin films (60 nm) on (001) SrTiO3 and (001) LaAlO3 substrates. Our experiments have also shown that a lattice strain may result in reduced Curie temperature and enhanced saturation magnetization. The spin glass like behavior is observed in the nanocrystalline thin films. The augmentation of the coercivity is possibly associated with nanocrystalline nature and the spin glass like phenomena in the films.

The roles of human serum IgA, in contrast to that of mucosal IgA, are relatively unexplored. Previous studies have shown that IgA mediates either pro- or anti-inflammatory effects in innate immune cells. Serum IgA has been shown to interact with many proteins and glycoproteins of which the functions and mechanisms are not fully characterized. Here, we present fresh perspectives into the roles of serum IgA, describing novel IgA–protein interactions, the importance of its glycosylation status in normal functions, and the plausible role of IgA as a driver and regulator of autoimmune diseases/immune overactivation. Other potential roles, including the regulation of cytokines, effector cell function, and homeostasis, are considered in view of the maintenance of immune function. We anticipate future research to uncover new anti-inflammatory or pro-inflammatory roles of human serum IgA in immune functions and dysfunctions, with implications on systemic lupus erythematosus (SLE).

The development of technologically important material zinc-blende ZnO has been hindered due to the difficulties inherent in obtaining a stable zinc-blende phase. In this paper, we fabricate the stable zinc-blende ZnO on Pt/Ti/SiO2/Si substrate through phase transformation from the originally wurtzite to the zinc-blende phase. X-ray diffraction data in combination with high-resolution TEM measurements provide the direct evidence on the formation of the well-defined zinc-blende structure with predominated (202) orientation. According to the experimental results and first principles calculations, the incorporation of titanium dopants into ZnO system favors the formation of the zinc-blende structure. The platinum (Pt) surface stabilizes the ZnO zinc-blende structure at the interface (thin film) due to its low ZnO/Pt interface energy, preventing the decomposition in ZnO wurtzite and Zn2TiO4. Additionally, magnetic and optical properties of the ZnO zinc-blende thin films are investigated. Unexpectedly, the film is found to exhibit magnetization of ~75 emu/cm3, while its ZnO wurtzite counterpart is non-ferromagnetic.

Cross-talk within the innate immune pathways is highly complex and contains many unknowns. Here, we discuss the different combinations of PAMPs, together with the sequence, order, and dosage of consecutive PAMP challenges, which determine the nature of the immune response by macrophages. The engagement of different Toll-like receptor (TLR) ligands leads to quantitatively and qualitatively unique cytokine production, showing that TLR pathway crosstalk enables the innate immune system to orchestrate immediate local and global responses. It is likely that multiple pathways are involved in the regulation of cytokine synergy, including many that have yet to be discovered.

The checkpoint between the life and death of macrophages is crucial for the host's frontline immune defense during acute phase infection. However, the mechanism as to how the immune cell equilibrates between apoptosis and immune response is unclear. Using in vitro and ex vivo approaches, we showed that macrophage survival is synchronized by SAG (sensitive to apoptosis gene), which is a key member of the ubiquitin–proteasome system (UPS). When challenged by pathogen-associated molecular patterns (PAMPs), we observed a reciprocal expression profile of pro- and antiapoptotic factors in macrophages. However, SAG knockdown disrupted this balance. Further analysis revealed that ubiquitination of Bax and SARM (sterile α- and HEAT/armadillo-motif-containing protein) by SAG-UPS confers survival advantage to infected macrophages. SAG knockdown caused the accumulation of proapoptotic Bax and SARM, imbalance of Bcl-2/Bax in the mitochondria, induction of cytosolic cytochrome c and activation of caspase-9 and -3, all of which led to disequilibrium between life and death of macrophages. In contrast, SAG-overexpressing macrophages challenged with PAMPs exhibited upregulation of protumorigenic cytokines (IL-1β, IL-6 and TNF-α), and downregulation of antitumorigenic cytokine (IL-12p40) and anti-inflammatory cytokine (IL-10). This suggests that SAG-dependent UPS is a key switch between immune defense and apoptosis or immune overactivation and tumorigenesis. Altogether, our results indicate that SAG-UPS facilitates a timely and appropriate level of immune response, prompting future development of potential immunomodulators of SAG-UPS.

Spin-transfer-torque magnetic random-access memory (STT-MRAM) devices are projected to find their applications in a wide range of systems such as portable devices which may be influenced by external magnetic fields. Therefore, it is important to study the magnetic immunity for the development of STT-MRAM immune to the data loss due to accidental or deliberate exposure to high magnetic fields. In this work, we propose a method to analyze the magnetic immunity of STT-MRAM arrays based on the bit error rate (BER) data in the presence of an external magnetic field. We propose an analytical equation to express the standard deviation of the energy barrier for the STT-MRAM array using a domain wall assisted switching model, which is confirmed using Monte Carlo simulations. We study 40 Mb BER data and predict that the STT-MRAM array can sustain a maximum field of 690 Oe and maintain BER below 0.1 ppm for ten-year exposure to the DC magnetic field at 85 °C.

Ordered Ni/Cu multilayered nanowires (NW's) were fabricated using the two bath method in which the AAO template was switched back and forth between the two baths each containing solutions of dissolved Ni and Cu sulfates repeatedly. Different combinations of periods in which templates spent in each bath were used. The SEM and TEM images of the NW's showed that the NW's were smooth and uniform. The VSM results showed that in the presence of a field applied parallel to the NW, the coercivity and squareness increased when the deposition times of the Ni and Cu increased. To account for the behavior when both the thicknesses of the Ni and Cu layers were increasing, the effects of the increase in the Cu layer partially offset the effects of the increase in the Ni layer. The highest coercivity and squareness achieved was 822 Oe and 0.949, respectively, when the deposition times were 8 min for the Ni deposition and 2.5 min for the Cu deposition.

Fe/SiO2 particles with core/shell structure were prepared by coating silica on the surface of a commercial spherical carbonyl iron via the hydrolysis process of tetraethyl orthosilicate (TEOS). The electromagnetic performance of commercial carbonyl iron and as-prepared Fe/SiO2 particles was studied theoretically and experimentally. As predicted by the theoretical calculation based on the Bruggeman formula and the Landau–Lifshitz–Gilbert (LLG) theory, the insulating surface layer of silica was effective to reduce the permittivity parameters of pure carbonyl iron. The measured results showed good agreement with the theoretical prediction. Although there was a little decrease in the permeability of the Fe/SiO2 core/shell particles, a better impedance match especially at higher frequency range was obtained when used as a microwave absorber. The reflection loss (RL) curves show that the lowest reflection loss of Fe/Epoxy composite (−20.5 GHz) was obtained corresponding to the frequency of 8.5 GHz when the thickness of the absorber was 3 mm. A different trend was observed in Fe/SiO2/Epoxy composite. The reflection loss value got lower by decreasing the thickness of absorbers. At the thickness of 2.2 mm, a relative low reflection loss (−17 GHz) corresponding to the frequency of 13.6 GHz was obtained. Compared with the Fe/Epoxy composite, the improvement on shifting the reflection loss peak to higher frequency and on reducing the optimal thickness of absorbers was made by Fe/SiO2/Epoxy composite.

For nearly five decades since its discovery, the role of natural IgG, which pre-exists in neonates and uninfected individuals, has remained unclear due to the general perception that natural antibodies lack affinity for pathogens. Here, we show for the first time that natural IgG recognizes a spectrum of bacteria through lectins like ficolin and mannose binding lectin (MBL). Infection-inflammation condition markedly increased the affinity of natural IgG for bacteria associated with ficolins. After opsonization with IgG:ficolin complex, the bacteria were phagocytosed by monocytes via FcγRI. Infection of C3−/− mice indicated that the natural IgG-mediated immune complex was formed independently of C3. AID−/− mice lacking IgG were susceptible to infection, unless reconstituted with natural IgG. Thus, we have proven that natural IgG is not quiescent; rather, it plays a vital and immediate role in immune defense. Our findings provide a fresh perspective on natural antibodies, opening new avenues to explore host–microbe interaction.