Increase in terahertz-wave intensity in a magnetic field due to difference-frequency mixing by exciton excitation in a GaAs/AlAs multiple quantum well.

Magnetic fields can increase the intensity of terahertz (THz) waves due to changing the dipole moment direction using the Lorentz force. This study reports the increase in the THz-wave intensity generated by differential frequency mixing using commercial permanent magnets under exciton-excitation. While a weak magnetic field applied to a multiple quantum well increases the THz-wave intensity due to excitons, a strong field causes its decrease. According to the calculations, the increase is caused by the electron-hole separation due to the Lorentz force. Furthermore, the calculations suggest the importance of carrier acceleration to enhance the intensity. Importantly, the increase in the THz-wave intensity due to differential frequency mixing does not require a strong magnetic field and can be achieved with inexpensive commercially available magnets.

Increase in terahertz-wave generation by difference frequency mixing by the overlap of exciton states in different GaAs/AlAs quantum wells and spectroscopic measurements.

Intense terahertz-wave emission in the higher frequency region can result in various applications such as terahertz spectroscopy and ultrafast data communication. In this study, an increase in terahertz waves by the overlap of exciton states in different quantum wells and spectroscopic demonstration are reported. The excitation energy dependence of signal intensity shows the effect of the overlap. The signals measured under the condition of square dependence of intensity on the excitation power indicate interference with the periods corresponding to the laser energy difference. Furthermore, the absorption coefficient of the transparent sheet is obtained at specific frequency. These results indicate that the generation of intense terahertz waves at various frequencies using excitons is possible and that difference frequency mixing is a useful terahertz-wave source.

Non-diffracting beam generated from a photonic integrated circuit based axicon-like lens.

We demonstrate an on-chip silicon-on-insulator (SOI) device to generate a non-diffracting beam of ≈850 µm length from a diffractive axicon-like lens etched using a low resolution (200 nm feature size, 250 nm gap) deep-ultraviolet lithographic fabrication. The device consists of circular gratings with seven stages of 1x2 multimode interferometers. We present a technique to apodize the gratings azimuthally by breaking up the circles into arcs which successfully increased the penetration depth in the gratings from ≈5 µm to ≈60 µm. We characterize the device's performance by coupling 1300±50 nm swept source laser in to the chip from the axicon and measuring the out-coupled light from a grating coupler. Further, we also present the implementation of balanced homodyne detection method for the spectral characterization of the device and show that the position of the output lobe of the axicon does not change significantly with wavelength.

Ex vivo expanded natural killer cells from breast cancer patients and healthy donors are highly cytotoxic against breast cancer cell lines and patient-derived tumours.

BACKGROUND: Natural killer (NK) cells play a critical role in cancer immunosurveillance. Recent developments in NK cell ex-vivo expansion makes it possible to generate millions of activated NK cells from a small volume of peripheral blood. We tested the functionality of ex vivo expanded NK cells in vitro against breast cancer cell lines and in vivo using a xenograft mouse model. The study aim was to assess functionality and phenotype of expanded NK cells from breast cancer patients against breast cancer cell lines and autologous primary tumours. METHODS: We used a well-established NK cell co-culture system to expand NK cells ex vivo from healthy donors and breast cancer patients and examined their surface marker expression. Moreover, we tested the ability of expanded NK cells to lyse the triple negative breast cancer and HER2-positive breast cancer cell lines MDA-MB-231 and MDA-MB-453, respectively. We also tested their ability to prevent tumour growth in vivo using a xenograft mouse model. Finally, we tested the cytotoxicity of expanded NK cells against autologous and allogeneic primary breast cancer tumours in vitro. RESULTS: After 3 weeks of culture we observed over 1000-fold expansion of NK cells isolated from either breast cancer patients or healthy donors. We also showed that the phenotype of expanded NK cells is comparable between those from healthy donors and cancer patients. Moreover, our results confirm the ability of ex vivo expanded NK cells to lyse tumour cell lines in vitro. While the cell lines examined had differential sensitivity to NK cell killing we found this was correlated with level of major histocompatibility complex (MHC) class I expression. In our in vivo model, NK cells prevented tumour establishment and growth in immunocompromised mice. Finally, we showed that NK cells expanded from the peripheral blood of breast cancer patients show high cytotoxicity against allogeneic and autologous patient-derived tumour cells in vitro. CONCLUSION: NK cells from breast cancer patients can be expanded similarly to those from healthy donors, have a high cytotoxic ability against breast cancer cell lines and patient-derived tumour cells, and can be compatible with current cancer treatments to restore NK cell function in cancer patients.

The acetylase/deacetylase couple CREB-binding protein/Sirtuin 1 controls hypoxia-inducible factor 2 signaling.

Hypoxia-inducible factors (HIFs) are oxygen-sensitive transcription factors. HIF-1α plays a prominent role in hypoxic gene induction. HIF-2α target genes are more restricted but include erythropoietin (Epo), one of the most highly hypoxia-inducible genes in mammals. We previously reported that HIF-2α is acetylated during hypoxia but is rapidly deacetylated by the stress-responsive deacetylase Sirtuin 1. We now demonstrate that the lysine acetyltransferases cAMP-response element-binding protein-binding protein (CBP) and p300 are required for efficient Epo induction during hypoxia. However, despite close structural similarity, the roles of CBP and p300 differ in HIF signaling. CBP acetylates HIF-2α, is a major coactivator for HIF-2-mediated Epo induction, and is required for Sirt1 augmentation of HIF-2 signaling during hypoxia in Hep3B cells. In comparison, p300 is a major contributor for HIF-1 signaling as indicated by induction of Pgk1. Whereas CBP can bind with HIF-2α independent of the HIF-2α C-terminal activation domain via enzyme/substrate interactions, p300 only complexes with HIF-2α through the C-terminal activation domain. Maximal CBP/HIF-2 signaling requires intact CBP acetyltransferase activity in both Hep3B cells as well as in mice.

Small signal modulation of photonic crystal surface emitting lasers.

We report the small-signal characterization of a PCSEL device, extracting damping factors and modulation efficiencies, and demonstrating -3 dB modulation bandwidths of up to 4.26 GHz. Based on modelling we show that, by reducing the device width and improving the active region design for high-speed modulation, direct modulation frequencies in excess of 50 GHz are achievable.

Hypoxia increases sirtuin 1 expression in a hypoxia-inducible factor-dependent manner.

Hypoxia-inducible factors (HIFs) are stress-responsive transcriptional regulators of cellular and physiological processes involved in oxygen metabolism. Although much is understood about the molecular machinery that confers HIF responsiveness to oxygen, far less is known about HIF isoform-specific mechanisms of regulation, despite the fact that HIF-1 and HIF-2 exhibit distinct biological roles. We recently determined that the stress-responsive genetic regulator sirtuin 1 (Sirt1) selectively augments HIF-2 signaling during hypoxia. However, the mechanism by which Sirt1 maintains activity during hypoxia is unknown. In this report, we demonstrate that Sirt1 gene expression increases in a HIF-dependent manner during hypoxia in Hep3B and in HT1080 cells. Impairment of HIF signaling affects Sirt1 deacetylase activity as decreased HIF-1 signaling results in the appearance of acetylated HIF-2α, which is detected without pharmacological inhibition of Sirt1. We also find that Sirt1 augments HIF-2 mediated, but not HIF-1 mediated, transcriptional activation of the isolated Sirt1 promoter. These data in summary reveal a bidirectional link of HIF and Sirt1 signaling during hypoxia.

Cu2O-Based Electrochemical Biosensor for Non-Invasive and Portable Glucose Detection.

Electrochemical voltammetric sensors are some of the most promising types of sensors for monitoring various physiological analytes due to their implementation as non-invasive and portable devices. Advantages in reduced analysis time, cost-effectiveness, selective sensing, and simple techniques with low-powered circuits distinguish voltammetric sensors from other methods. In this work, we developed a Cu2O-based non-enzymatic portable glucose sensor on a graphene paste printed on cellulose cloth. The electron transfer of Cu2O in a NaOH alkaline medium and sweat equivalent solution at very low potential (+0.35 V) enable its implementation as a low-powered portable glucose sensor. The redox mechanism of the electrodes with the analyte solution was confirmed through cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy studies. The developed biocompatible, disposable, and reproducible sensors showed sensing performance in the range of 0.1 to 1 mM glucose, with a sensitivity of 1082.5 ± 4.7% µA mM-1 cm-2 on Cu2O coated glassy carbon electrode and 182.9 ± 8.83% µA mM-1 cm-2 on Cu2O coated graphene printed electrodes, making them a strong candidate for future portable, non-invasive glucose monitoring devices on biodegradable substrates. For portable applications we demonstrated the sensor on artificial sweat in 0.1 M NaOH solution, indicating the Cu2O nanocluster is selective to glucose from 0.0 to +0.6 V even in the presence of common interference such as urea and NaCl.

HDACi promotes inflammatory remodeling of the tumor microenvironment to enhance epitope spreading and antitumor immunity.

Adoptive cell therapy (ACT) with tumor-specific memory T cells has shown increasing efficacy in regressing solid tumors. However, tumor antigen heterogeneity represents a longitudinal challenge for durable clinical responses due to the therapeutic selective pressure for immune escape variants. Here, we demonstrated that delivery of the class I histone deacetylase inhibitor MS-275 promoted sustained tumor regression by synergizing with ACT in a coordinated manner to enhance cellular apoptosis. We found that MS-275 altered the tumor inflammatory landscape to support antitumor immunoactivation through the recruitment and maturation of cross-presenting CD103+ and CD8+ DCs and depletion of Tregs. Activated endogenous CD8+ T cell responses against nontarget tumor antigens were critically required for the prevention of tumor recurrence. Importantly, MS-275 altered the immunodominance hierarchy by directing epitope spreading toward the endogenous retroviral tumor-associated antigen p15E. Our data suggest that MS-275 in combination with ACT multimechanistically enhanced epitope spreading and promoted long-term clearance of solid tumors.

CLIC-01: Manufacture and distribution of non-cryopreserved CAR-T cells for patients with CD19 positive hematologic malignancies.

Access to commercial CD19 CAR-T cells remains limited even in wealthy countries like Canada due to clinical, logistical, and financial barriers related to centrally manufactured products. We created a non-commercial academic platform for end-to-end manufacturing of CAR-T cells within Canada's publicly funded healthcare system. We report initial results from a single-arm, open-label study to determine the safety and efficacy of in-house manufactured CD19 CAR-T cells (entitled CLIC-1901) in participants with relapsed/refractory CD19 positive hematologic malignancies. Using a GMP compliant semi-automated, closed process on the Miltenyi Prodigy, T cells were transduced with lentiviral vector bearing a 4-1BB anti-CD19 CAR transgene and expanded. Participants underwent lymphodepletion with fludarabine and cyclophosphamide, followed by infusion of non-cryopreserved CAR-T cells. Thirty participants with non-Hodgkin's lymphoma (n=25) or acute lymphoblastic leukemia (n=5) were infused with CLIC-1901: 21 males (70%), median age 66 (range 18-75). Time from enrollment to CLIC-1901 infusion was a median of 20 days (range 15-48). The median CLIC-1901 dose infused was 2.3 × 106 CAR-T cells/kg (range 0.13-3.6 × 106/kg). Toxicity included ≥ grade 3 cytokine release syndrome (n=2) and neurotoxicity (n=1). Median follow-up was 6.5 months. Overall response rate at day 28 was 76.7%. Median progression-free and overall survival was 6 months (95%CI 3-not estimable) and 11 months (95% 6.6-not estimable), respectively. This is the first trial of in-house manufactured CAR-T cells in Canada and demonstrates that administering fresh CLIC-1901 product is fast, safe, and efficacious. Our experience may provide helpful guidance for other jurisdictions seeking to create feasible and sustainable CAR-T cell programs in research-oriented yet resource-constrained settings. Clinical trial registration: https://clinicaltrials.gov/ct2/show/NCT03765177, identifier NCT03765177.

Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro.

Although natural killer (NK) cells have become a promising immune effector cell for chimeric antigen receptor (CAR)-based therapy, generating human CAR-NK cells with high transgene efficiency has been challenging. In this protocol, we describe how to generate CAR-NK cells with transduction efficiencies >15% from healthy donor ex vivo expanded NK cells using third generation lentiviral vectors (LVs). We also show how to assess CAR-NK cell anti-tumor function in vitro using a flow cytometry-based killing assay. For complete details on the use and execution of this protocol, please refer to Portillo et al. (2021).

Hepatic FGF21 preserves thermoregulation and cardiovascular function during bacterial inflammation.

Sickness behaviors, including anorexia, are evolutionarily conserved responses to acute infections. Inflammation-induced anorexia causes dramatic metabolic changes, of which components critical to survival are unique depending on the type of inflammation. Glucose supplementation during the anorectic period induced by bacterial inflammation suppresses adaptive fasting metabolic pathways, including fibroblast growth factor 21 (FGF21), and decreases survival. Consistent with this observation, FGF21-deficient mice are more susceptible to mortality from endotoxemia and polybacterial peritonitis. Here, we report that increased circulating FGF21 during bacterial inflammation is hepatic derived and required for survival through the maintenance of thermogenesis, energy expenditure, and cardiac function. FGF21 signaling downstream of its obligate coreceptor, ß-Klotho (KLB), is required in bacterial sepsis. However, FGF21 modulates thermogenesis and chronotropy independent of the adipose, forebrain, and hypothalamus, which are operative in cold adaptation, suggesting that in bacterial inflammation, either FGF21 signals through a novel, undescribed target tissue or concurrent signaling of multiple KLB-expressing tissues is required.

Expanded human NK cells armed with CAR uncouple potent anti-tumor activity from off-tumor toxicity against solid tumors.

Despite the remarkable success of chimeric antigen receptor (CAR)-T cells against hematologic malignancies, severe off-tumor effects have constrained their use against solid tumors. Recently, CAR-engineered natural killer (NK) cells have emerged as an effective and safe alternative. Here, we demonstrate that HER2 CAR-expression in NK cells from healthy donors and patients with breast cancer potently enhances their anti-tumor functions against various HER2-expressing cancer cells, regardless of MHC class I expression. Moreover, HER2 CAR-NK cells exert higher cytotoxicity than donor-matched HER2 CAR-T cells against tumor targets. Importantly, unlike CAR-T cells, HER2 CAR-NK cells do not elicit enhanced cytotoxicity or inflammatory cytokine production against non-malignant human lung epithelial cells with basal HER2 expression. Further, HER2 CAR-NK cells maintain high cytotoxic function in the presence of immunosuppressive factors enriched in solid tumors. These results show that CAR-NK cells may be a highly potent and safe source of immunotherapy in the context of solid tumors.

Broadband THz absorption spectrometer based on excitonic nonlinear optical effects.

A broadly tunable THz source is realized via difference frequency generation, in which an enhancement to χ(3) that is obtained via resonant excitation of III-V semiconductor quantum well excitons is utilized. The symmetry of the quantum wells (QWs) is broken by utilizing the built-in electric-field across a p-i-n junction to produce effective χ(2) processes, which are derived from the high χ(3). This χ(2) media exhibits an onset of nonlinear processes at ~4 W cm-2, thereby enabling area (and, hence, power) scaling of the THz emitter. Phase matching is realized laterally through normal incidence excitation. Using two collimated 130 mW continuous wave (CW) semiconductor lasers with ~1-mm beam diameters, we realize monochromatic THz emission that is tunable from 0.75 to 3 THz and demonstrate the possibility that this may span 0.2-6 THz with linewidths of ~20 GHz and efficiencies of ~1 × 10-5, thereby realizing ~800 nW of THz power. Then, transmission spectroscopy of atmospheric features is demonstrated, thereby opening the way for compact, low-cost, swept-wavelength THz spectroscopy.

Alveolar Macrophage Chemokine Secretion Mediates Neutrophilic Lung Injury in Nox2-Deficient Mice.

Acute lung injury (ALI), developing as a component of the systemic inflammatory response syndrome (SIRS), leads to significant morbidity and mortality. Reactive oxygen species (ROS), produced in part by the neutrophil NADPH oxidase 2 (Nox2), have been implicated in the pathogenesis of ALI. Previous studies in our laboratory demonstrated the development of pulmonary inflammation in Nox2-deficient (gp91phox-/y) mice that was absent in WT mice in a murine model of SIRS. Given this finding, we hypothesized that Nox2 in a resident cell in the lung, specifically the alveolar macrophage, has an essential anti-inflammatory role. Using a murine model of SIRS, we examined whole-lung digests and bronchoalveolar lavage fluid (BALf) from WT and gp91phox-/y mice. Both genotypes demonstrated neutrophil sequestration in the lung during SIRS, but neutrophil migration into the alveolar space was only present in the gp91phox-/y mice. Macrophage inflammatory protein (MIP)-1α gene expression and protein secretion were higher in whole-lung digest from uninjected gp91phox-/y mice compared to the WT mice. Gene expression of MIP-1α, MCP-1, and MIP-2 was upregulated in alveolar macrophages obtained from gp91phox-/y mice at baseline compared with WT mice. Further, ex vivo analysis of alveolar macrophages, but not bone marrow-derived macrophages or peritoneal macrophages, demonstrated higher gene expression of MIP-1α and MIP-2. Moreover, isolated lung polymorphonuclear neutrophils migrate to BALf obtained from gp91phox-/y mice, further providing evidence of a cell-specific anti-inflammatory role for Nox2 in alveolar macrophages. We speculate that Nox2 represses the development of inflammatory lung injury by modulating chemokine expression by the alveolar macrophage.

Ex Vivo-expanded Natural Killer Cells Derived From Long-term Cryopreserved Cord Blood are Cytotoxic Against Primary Breast Cancer Cells.

With over 600,000 units of umbilical cord blood (CB) stored on a global scale, it is important to elucidate the therapeutic abilities of this cryopreserved reservoir. In the advancing field of natural killer (NK) cell cancer immunotherapy, CB has proven to be a promising and noninvasive source of therapeutic NK cells. Although studies have proven the clinical efficacy of using long-term cryopreserved CB in the context of hematopoietic stem cell transplantations, little is known about its use for the ex vivo expansion of effector immune cells. Therefore, our group sought to derive ex vivo-expanded NK cells from long-term cryopreserved CB, using an artificial antigen presenting cell-mediated expansion technique. We compared the expansion potential and antitumor effector function of CB-derived NK (CB-NK) cells expanded from fresh (n=4), short-term cryopreserved (<1-year old, n=5), and long-term cryopreserved (1-10-year old, n=5) CB. Here, we demonstrated it is possible to obtain an exponential amount of expanded CB-NK cells from long-term cryopreserved CB. Ex vivo-expanded CB-NK cells had an increased surface expression of activating markers and showed potent antitumor function by producing robust levels of proinflammatory cytokines, interferon-γ, and tumor necrosis factor-α. Moreover, expanded CB-NK cells (n=3-5) demonstrated cytotoxicity towards primary breast cancer cells (n=2) derived from a triple-negative breast cancer and an estrogen receptor-positive/progesterone receptor-positive breast cancer patient. Long-term cryopreservation had no effect on the expansion potential or effector function of expanded CB-NK cells. Therefore, we propose that long-term cryopreserved CB remains clinically useful for the ex vivo expansion of therapeutic NK cells.

Near-infrared and mid-infrared semiconductor broadband light emitters.

Semiconductor broadband light emitters have emerged as ideal and vital light sources for a range of biomedical sensing/imaging applications, especially for optical coherence tomography systems. Although near-infrared broadband light emitters have found increasingly wide utilization in these imaging applications, the requirement to simultaneously achieve both a high spectral bandwidth and output power is still challenging for such devices. Owing to the relatively weak amplified spontaneous emission, as a consequence of the very short non-radiative carrier lifetime of the inter-subband transitions in quantum cascade structures, it is even more challenging to obtain desirable mid-infrared broadband light emitters. There have been great efforts in the past 20 years to pursue high-efficiency broadband optical gain and very low reflectivity in waveguide structures, which are two key factors determining the performance of broadband light emitters. Here we describe the realization of a high continuous wave light power of >20 mW and broadband width of >130 nm with near-infrared broadband light emitters and the first mid-infrared broadband light emitters operating under continuous wave mode at room temperature by employing a modulation p-doped InGaAs/GaAs quantum dot active region with a 'J'-shape ridge waveguide structure and a quantum cascade active region with a dual-end analogous monolithic integrated tapered waveguide structure, respectively. This work is of great importance to improve the performance of existing near-infrared optical coherence tomography systems and describes a major advance toward reliable and cost-effective mid-infrared imaging and sensing systems, which do not presently exist due to the lack of appropriate low-coherence mid-infrared semiconductor broadband light sources.

Splenectomy impairs diffusive oxygen transport in the lung of dogs.

The spleen acts as an erythrocyte reservoir in highly aerobic species such as the dog and horse. Sympathetic-mediated splenic contraction during exercise reversibly enhances convective O2 transport by increasing hematocrit, blood volume, and O2-carrying capacity. Based on theoretical interactions between erythrocytes and capillary membrane (Hsia CCW, Johnson RL Jr, and Shah D. J Appl Physiol 86: 1460-1467, 1999) and experimental findings in horses of a postsplenectomy reduction in peripheral O2-diffusing capacity (Wagner PD, Erickson BK, Kubo K, Hiraga A, Kai M, Yamaya Y, Richardson R, and Seaman J. Equine Vet J 18, Suppl: 82-89, 1995), we hypothesized that splenic contraction also augments diffusive O2 transport in the lung. Therefore, we have measured lung diffusing capacity (DL(CO)) and its components during exercise by a rebreathing technique in six adult foxhounds before and after splenectomy. Splenectomy eliminated exercise-induced polycythemia, associated with a 30% reduction in maximal O2 uptake. At any given pulmonary blood flow, DL(CO) was significantly lower after splenectomy owing to a lower membrane diffusing capacity, whereas pulmonary capillary blood volume changed variably; microvascular recruitment, indicated by the slope of the increase in DL(CO) with respect to pulmonary blood flow, was also reduced. We conclude that splenic contraction enhances both convective and diffusive O2 transport and provides another compensatory mechanism for maintaining alveolar O2 transport in the presence of restrictive lung disease or ambient hypoxia.

GaAs-Based Superluminescent Light-Emitting Diodes with 290-nm Emission Bandwidth by Using Hybrid Quantum Well/Quantum Dot Structures.

A high-performance superluminescent light-emitting diode (SLD) based upon a hybrid quantum well (QW)/quantum dot (QD) active element is reported and is assessed with regard to the resolution obtainable in an optical coherence tomography system. We report on the appearance of strong emission from higher order optical transition from the QW in a hybrid QW/QD structure. This additional emission broadening method contributes significantly to obtaining a 3-dB linewidth of 290 nm centered at 1200 nm, with 2.4 mW at room temperature.

Optical characterization of In-flushed InAs/GaAs quantum dots emitting a broadband spectrum with multiple peaks at ~1 µm.

We investigated optical properties of In-flushed InAs quantum dots (QDs) grown on a GaAs substrate by molecular beam epitaxy. By using the In-flush technique for setting the height of self-assembled InAs QDs, we have tuned the emission wavelength of InAs QDs to the ~1 µm regime, which can be utilized as a non-invasive and deeply penetrative probe for biological and medical imaging systems. The controlled emission exhibited a broadband spectrum comprising multiple peaks with an interval of approximately 30 meV. We examined the origin of the multiple peaks using spectral and time-resolved photoluminescence, and concluded that it is attributed to monolayer step fluctuations in the height of the In-flushed QDs. This feature can be advantageous for realizing a broadband light source centered at the ~1 µm regime, which is especially suitable for the non-invasive cross-sectional biological and medical imaging system known as optical coherence tomography.