Size Expansion Research Articles

How does green finance influence industrial green total factor productivity? Empirical research from China

Industry is the pillar of the real economy, and its green development contributes to achieving the green and low-carbon development goals proposed by the United Nations. This paper constructs an analytical framework of "green finance-green technological innovation-industrial green total factor productivity" to study how green finance (GF) influences industrial green total factor productivity (IGTFP). Based on the panel data from 30 provinces and 281 cities in China between 2011–2019，this study measures the GF levels and the IGTFP index by using the entropy method and super-efficiency slacks-based model with Malmquist-Luenberger index method, to conduct a series of empirical tests. Also, we focus on the mechanism effect of green technology innovation (GTI) in the process of GF affecting IGTFP. Additionally, we have discussed the supporting role of digital financial inclusion (DFI) in green finance development at the micro level. The main findings include: (1) The development of green finance (GF) can significantly improve IGTFP, and it’s still credible after a series of robustness tests. (2) The heterogeneity analysis indicates that the influence of GF on the IGTFP varies from different cities, and this positive effect tends to increase with the expansion of city size. (3) Mechanism tests show that green technological innovation (GTI) has a positive mechanism effect in the process of GF influencing IGTFP. (4) The discussion indicates that digital financial inclusion can support green finance development, so as to promote industrial green development in China. This study has momentous theoretical value and policy implications for China and other developing countries in exploring green finance and industrial green development.

The Interactive Effects of Communication Network Structure and Organizational Size on Task Performance in Project-Based Organizations: The Mediating Role of Bootleg Innovation Behavior

A PBO is a temporary organization formed by assembling members with diverse experiences and backgrounds, aimed at achieving specific innovation goals. Constructing a reasonable communication network structure and enhancing organizational synergy are effective ways to promote the sustainable development of the system. This study, based on the Input–Process–Output (IPO) model and social network analysis, utilized a group collaboration platform to conduct a three-stage communication experiment on 685 construction project managers. Under two organizational sizes, the internal mechanism of how communication networks with two levels of centralization influence task performance were tested. The results indicate that in the case of a smaller organizational size, PBOs using a decentralized communication network tend to achieve higher task performance. However, as the organizational size expands, PBOs employing a centralized communication network may surpass in task performance. Additionally, we found that with the expansion of organizational size, bootleg innovation behaviors of organizational members are continually stimulated, further enhancing collective task performance. This study, based on the evolution of communication network parameters, explores the structural characteristics of organizational communication networks and the mechanisms underlying the emergence of bootleg innovation behaviors. It delineates the key pathways for improving collective task performance. The findings can provide a scientific reference for the organizational evolution and development of engineering project management.

Go west: Population genomics reveals unexpected population fluctuations and little gene flow in Western hemisphere populations of the predatory lady beetle, Hippodamia convergens.

Hippodamia convergens-the convergent lady beetle, has been used extensively in augmentative biological control of aphids, thrips, and whiteflies across its native range in North America, and was introduced into South America in the 1950s. Overwintering H. convergens populations from its native western range in the United States are commercially collected and released across its current range in the eastern USA, with little knowledge of the effectiveness of its augmentative biological control. Here we use a novel ddRADseq-based SNP/haplotype discovery approach to estimate its range-wide population diversity, differentiation, and recent evolutionary history. Our results indicate (1) significant population differentiation among eastern USA, western USA, and South American populations of H. convergens, with (2) little to no detectable recent admixture between them, despite repeated population augmentation, and (3) continued recent population size expansion across its range. These results contradict previous findings using microsatellite markers. In light of these new findings, the implications for the effectiveness of augmentative biological control using H. convergens are discussed. Additionally, because quantifying the non-target effects of augmentative biological control is a difficult problem in migratory beetles, our results could serve as a cornerstone in improving and predicting the efficacy of future releases of H. convergens across its range.

Вроджені кісти яєчника: діагностика та лікування в неонатальному періоді

The indications and method of surgery in newborns with prenatally diagnosed ovarian cysts remain under discussion. Purpose - to summarize own experience in the treatment of newborns with congenital ovarian cysts. Materials and methods. This study based on the results of treatment of 15 newborns that were operated in I surgical department of Lviv Regional Children’s Clinical Hospital «OKHMATDYT» at 2010-2021 years. For the diagnose confirming, the US was applied in all newborns after birth. The urgent hospitalization, immediately after birth, required 7 (46.7%) newborns and 4 (26.7%) newborns - during of the first month of life, by that 4 (26.7%) infants were hospitalized for elective surgery during six months after birth. The open surgery was applied in 9 (60%) patients and laparoscopic - in 6 (40%) patients. The postoperative mortality was 6.7%. Results of the study were evaluated by the statistical program StatPlus: mac, AnalystSoft Inc. (version v8). Results. Among of 54 fetuses, after the birth the presence of cyst was confirmed in 15 (27.8%) of newborns, which were operated. Indications for urgent surgery immediately after birth (46.7% of newborns) was the presence of huge (>6 cm) cyst in abdomen, which in 2 newborns determined with respiratory disorders and in 2 newborns - intestinal obstruction. Increasing of the cyst’s size with the pain syndrome was the indication for the urgent surgery during of the first month of life (26.7% of infants) and the presence of cyst without tendency to decreasing of its size was the indication for elective surgery (26.7% of infants). Ovary-sparing surgery was succeeded only in 20% of patients and in 80% of patients performed adnexectomy. Conclusions. By the antenatally revealing of ovarian cyst, the scheduled US control is required with the aim to monitoring of the dynamic of cyst size. The indications for the surgery in neonatal period are: 1) cysts with the diameter >4 cm; 2) clear suspicions for the complications (torsion, self-amputation, compression of abdominal organs); 3) enlargement of cyst size after the born. The choice of method of surgery determined by size and the character of cystic formation. The research was carried out in accordance with principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of clinical hospital «OKHMATDYT». The informed agreement of parents was obtained for conducting the study. No conflict of interests was declared by the authors.

Coarse-Grained Monte Carlo Simulations with Octree Cells for Geopolymer Nucleation at Different pH Values.

Geopolymers offer a potential alternative to ordinary Portland cement owing to their performance in mechanical and thermal properties, as well as environmental benefits stemming from a reduced carbon footprint. This paper endeavors to build upon prior atomistic computational work delving deeper into the intricate relationship between pH levels and the resulting material's properties, including pore size distribution, geopolymer nucleate cluster dimensions, total system energy, and monomer poly-condensation behavior. Coarse-grained Monte Carlo (CGMC) simulation inputs include tetrahedral geometry and binding energy parameters derived from DFT simulations for aluminate and silicate monomers. Elevated pH values may can alter reactivity and phase stability, or, in the structural concrete application, may passivate the embedded steel reinforcement. Thus, we examine the effects of pH values set at 11, 12, and 13 (based on silicate speciation chemistry), investigating their respective contributions to the nucleation of geopolymers. To simulate a larger system to obtain representative results, we propose the numerical implementation of an Octree cell. Finally, we further digitize the resulting expanded structure to ascertain pore size distribution, facilitating a comparative analysis. The novelty of this study is underscored by its expansion in both system size, more accurate monomer representation, and pH range when compared to previous CGMC simulation approaches. The results unveil a discernible correlation between the number of clusters and pores under specific pH levels. This links geopolymerization mechanisms under varying pH conditions to the resulting chemical properties and final structural state.

Study on flow characteristics of exhaust shaft with parallel fans in high-rise residential buildings

The exhaust shaft is often used to centrally extract cooking fumes captured by parallel fans on each floor of high-rise buildings. Inadequate airflow rate and excessive pressure in the exhaust shaft lead to poor ventilation and unsatisfactory air quality in kitchens. This issue is made worse by mixed installations of exhaust fans with different dynamic performances. In this study, the flow characteristics in the exhaust shaft with parallel fans are investigated by full-scale experiments and computational fluid dynamics (CFD). Based on the orthogonal experimental design, the cross-sectional area, concurrent operating rate, operating position distribution, and P-Q curves are key factors affecting the flow rate distribution significantly. Mixed installation of fans deteriorates the uniformity of the flow distribution along the shaft by even 62.6%. The competition for flow rate between different types of fans is obeying the Matthew effect. Concentration at the bottom and more simultaneous operations exacerbate this problem. An 28% enlargement in the shaft size increases the flow rate by 28.9% and improves the uniformity of the flow distribution by 50%. The stack effect is positive and unstable, with up to a 6.49% increase in system flow rate as a function of the cooking heat intensity. This work can provide guidance for the design of centralized exhaust systems in high-rise residential buildings.

Comparative Performance Assessment of Sizing of Electric Motor through Analytical Approach for Electric Vehicle Application

For battery electric vehicle (BEV), electric motor is the only power source. The motor drive system in electric vehicle has many challenges like cost, weight, efficiency, detail torque speed characteristics, motor power rating. Accurate motor power rating prediction is very much necessary to fulfil the performance requirement of electric vehicle. Selection of oversized motor for Electric vehicle application results into overprice motor, more energy consumption and decline in vehicle range. Due to this there will expansion of battery size and increases the overall cost of vehicle. On the other hand, selection of undersized motor leads to poor vehicle performance that limits the drivability of vehicle. This paper gives detailed calculations of motor rating of electric motor. New approach is developed to predict the correct power rating of motor without affecting the desired vehicle performance. According to the requirement, accurate prediction of power rating of motor has been taken for cost efficient and upgradation the performance of motor. The work presented in this paper adopts a method of vehicle performance assessment on the basis of comparison with four different methods with new developed method. Traction motor is major & most expensive component of EV. Selection of motor play’s crucial role in the development of electric vehicle. Undersized and oversized motor results in rise in cost of motor which ultimately increases the cost of vehicle. But by using appropriate motor this cost been reduced up to 50-60% with performance enhancement.

Genomic analysis of lineage-specific transmission of multidrug resistance tuberculosis in China

ABSTRACT Objectives We investigated the genetic diversities and lineage-specific transmission dynamics of multidrug-resistant tuberculosis (MDR-TB), with the goal of determining the potential factors driving the MDR epidemics in China. Methods We curated a large nationwide Mycobacterium tuberculosis (M. tuberculosis) whole genome sequence data set, including 1313 MDR strains. We reconstructed the phylogeny and mapped the transmission networks of MDR-TB across China using Bayesian inference. To identify drug-resistance variants linked to enhanced transmissibility, we employed ordinary least-squares (OLS) regression analysis. Result The majority of MDR-TB strains in China belong to lineage 2.2.1. Transmission chain analysis has indicated that the repeated and frequent transmission of L2.2.1 plays a central role in the establishment of MDR epidemic in China, but no occurrence of a large predominant MDR outbreak was detected. Using OLS regression, the most common single nucleotide polymorphisms (SNPs) associated with resistance to isoniazid (katG_p.Ser315Thr and katG_p.Ser315Asn) and rifampicin (rpoB_p.Ser450Leu, rpoB_p.His445Tyr, rpoB_p.His445Arg, rpoB_p.His445Asp, and rpoB_p.His445Asn) were more likely to be found in L2 clustered strains. Several putative compensatory mutations in rpoA, rpoC, and katG were significantly associated with clustering. The eastern, central, and southern regions of China had a high level of connectivity for the migration of L2 MDR strains throughout the country. The skyline plot showed distinct population size expansion dynamics for MDR-TB lineages in China. Conclusion MDR-TB epidemic in China is predominantly driven by the spread of highly transmissible Beijing strains. A range of drug-resistance mutations of L2 MDR-TB strains displayed minimal fitness costs and may facilitate their transmission.

PSO‐GA‐SVRmodel for S‐parameters of radio‐frequency power amplifier under different temperatures

AbstractSupport vector machine (SVR) has been introduced into the modeling of S‐parameters in radio‐frequency (RF) power amplifiers (PA). The modeling accuracy and speed of SVR are primarily affected by the penalty parametercand the kernel function coefficientγ. Using the traditional grid search technique to determine these two parameters is time‐consuming and labor‐intensive, and ensuring the model's accuracy is not easy. This article proposes an S‐parameters modeling method based on PSO‐GA‐SVR to improve the SVR's modeling accuracy and speed. The model mainly focuses on particle swarm optimization (PSO) and combines selection, crossover, and mutation operations in genetic algorithms (GA). The fitness values are arranged from small to large in each iteration process, and the first 1/3 are selected for crossover and mutation. Then, the resulting new particle swarms are introduced into the original particle swarm population for searching. On the one hand, PSO‐GA extends the population size and reduces the possibility of falling into local optimization. On the other hand, due to population size expansion, the number of iteration rounds is reduced, and the modeling speed is also increased. The experimental results show that compared to SVR, GA‐SVR, and PSO‐SVR, the proposed PSO‐GA‐SVR can improve the modeling accuracy by more than one magnitude or more while also increasing modeling speed by one magnitude or more. Furthermore, compared with the classical machine learning algorithms such as gradient boosting, random forest, and gcForset, the proposed PSO‐GA‐SVR improves the modeling accuracy by one order of magnitude and the modeling speed by two orders of magnitude more than gradient boosting, random forest, and improves the modeling speed by one order of magnitude more than gcForset.

Nectin-4 regulates cellular senescence-associated enlargement of cell size

Cellular senescence is defined as irreversible growth arrest induced by various stress, such as DNA damage and oxidative stress. Senescent cells exhibit various characteristic morphological changes including enlarged morphology. In our recent study, we identified Nectin-4 to be upregulated in cellular senescence by comparative transcriptomic analysis. However, there are few reports on the relationship between Nectin-4 and senescence. Therefore, we analyzed the function of Nectin-4 in senescence and its biological significance. When overexpressed with Nectin-4, the cells exhibited the enlarged cell morphology closely resembling senescent cells. In addition, the cell size enlargement during DNA damage-induced senescence was suppressed by knockdown of Nectin-4, while there were no significant changes in senescence induction. These results suggest that Nectin-4 is not involved in the regulation of senescence itself but contributes to the senescence-associated cell size increase. Furthermore, the Nectin-4-dependent cell size increase was found to be mediated by Src family kinase (SFK)/PI3 kinase (PI3K)/Rac1 pathway. To explore the functional consequences of cell size enlargement, we analyzed cell survival in Nectin-4-depleted senescent cells. Single-cell tracking experiments revealed that Nectin-4 knockdown induced apoptosis in senescent cells, and there is a strong positive correlation between cell size and survival rate. These results collectively indicate that Nectin-4 plays a causative role in the senescence-associated cell size enlargement via SFK/PI3K/Rac1, which can contribute to survival of senescent cells.

Acidity’s Impact on Yield, Morphological Structure, and Surface Functionalities of Polyaniline Synthesised via Oxidative Polymerisation

This research aimed to examine how varying the acidity levels during the chemical oxidative polymerisation (COP) of aniline affects the colour, yield, morphology, and surface functionalities of polyaniline (PANI). In the COP method, aniline monomer was first dissolved in hydrochloric acid (HCl) medium; the solution was then drop added with ammonium persulfate to initiate the polymerisation process. To answer the research objective, the medium acidity was manipulated using different concentrations of HCl (i.e., 0 M, 0.01 M, 0.1 M, 1 M and 10 M). The as-synthesised PANIs were then purified and characterised for their physicochemical properties. The findings revealed that reducing the medium pH from 2.35 (in distilled water) to 0.80 (in 10 M HCl) resulted in a colour change from light brownish to dark green, indicating the formation of emeraldine PANI. Furthermore, it was observed that the amount of PANI yield is inversely related to the medium pH. A decrease in medium pH from 2.35 to 0.80 remarkably increased the PANI yield by 3.2 times. Regarding morphological structure, higher medium acidity produced PANI with well-defined globule shapes, while lower HCl concentrations resulted in a mixed morphology including globules, tubes, and plate-like structures. In addition, it was found that the diameter of globule-like PANI has increased from ~266.67 nm to ~508.33 nm, when the HCl concentration was changed from 0 M to 10 M. Apparently, increasing the acidity of the COP medium promotes enlargement of the PANI size. Despite that, there is not much change in the surface functional groups of the formed PANI. Overall, this study offers insights into manipulating PANI properties by adjusting the medium acidity during the COP process. While the impacts of medium acidity on yield, morphology, and functionalities of PANI were revealed, further investigation is required to evaluate the effect on electrical properties.

Treatment with a new barbituric acid derivative suppresses diet-induced metabolic dysfunction and non-alcoholic fatty liver disease in mice

IntroductionNonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease, often accompanied by obesity, diabetes, and increased risks of depression and anxiety. Currently, there are no FDA-approved drugs to treat NAFLD and its related systemic symptoms. Previously, we identified a new barbituric acid derivative (BA-5) that expressed effectiveness against fibrosis and drug-resistant hepatocellular carcinoma. AimsThis study investigated the potential of BA-5 against high-fat diet (HFD)-induced NAFLD and mood disorders in mice. Main methodsSix-weeks-old male C57BL/6 mice were fed with a 45 % HFD for 8 weeks to induce NAFLD and associated metabolic disorders. Mice were treated with a BA-5 and the therapeutic effects and the underlying molecular mechanisms were investigated. Key findingsAdministration of BA-5 significantly reduced serum levels of alanine aminotransferase (ALT), low-density lipoprotein (LDL), fatty acids (FA), and triglycerides (TG) in HFD-fed mice. BA-5 treatment decreased expressions of hepatic lipogenesis-related markers (acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and ATP-citrate lyase (ACLY)), increased fatty acid oxidation markers (carnitine palmitoyltransferase 1A (CPT1A) and acyl-CoA oxidase 1 (ACOX1)), and attenuated hepatic fat accumulation in HFD-fed mice. Moreover, HFD-induced adipocyte size enlargement and activation of lipolysis markers such as phosphorylated (p)-hormone-sensitive lipase (HSL) 565, p-HSL 660, and perilipin were inhibited in BA-5-treated mice. Notably, HFD-induced anxiety- and depression-like behaviors significantly improved in the BA-5 treated group through enhanced anti-inflammatory responses in the hippocampus. SignificanceThis study provides new insights into clinical therapeutic strategies of barbituric acid derivatives for HFD-induced NAFLD and associated mood disturbances.

Neutron dosimetry with a pair of TLDs for the Elekta Precise medical linac and the evaluation of optimum moderator thickness for the conversion of fast to thermal neutrons

IntroductionIn this study, TLD 600 and TLD 700 pairs were used to measure the neutron dose of Elekta Precise medical linac. To this end, the optimum moderate thickness for the conversion of fast to thermal neutrons were evaluated. Materials and methods241Am-Be and 252Cf sources were simulated to calculate the optimum thicknesses of the moderator for the conversion of maximum fast neutrons (FN) into thermal neutrons (TN). Pair TLDs were used to measure F&TN doses for three different field sizes at four depths of the medical linac. ResultsThe maximum thickness of the moderator was optimized at 6 cm. The measurement results demonstrated that the TN dose increased with the expansion of field size and depth. The FN dose, which was converted TN, exhibits behaviors comparable to the TN due to its nature. ConclusionThis study presents the optimum thickness for the moderator to convert FN into TN and measure F&TN using TLDs.

The evolution of human altriciality and brain development in comparative context.

Human newborns are considered altricial compared with other primates because they are relatively underdeveloped at birth. However, in a broader comparative context, other mammals are more altricial than humans. It has been proposed that altricial development evolved secondarily in humans due to obstetrical or metabolic constraints, and in association with increased brain plasticity. To explore this association, we used comparative data from 140 placental mammals to measure how altriciality evolved in humans and other species. We also estimated how changes in brain size and gestation length influenced the timing of neurodevelopment during hominin evolution. Based on our data, humans show the highest evolutionary rate to become more altricial (measured as the proportion of adult brain size at birth) across all placental mammals, but this results primarily from the pronounced postnatal enlargement of brain size rather than neonatal changes. In addition, we show that only a small number of neurodevelopmental events were shifted to the postnatal period during hominin evolution, and that they were primarily related to the myelination of certain brain pathways. These results indicate that the perception of human altriciality is mostly driven by postnatal changes, and they point to a possible association between the timing of myelination and human neuroplasticity.

Study of the Optical Properties of 3MPA CdTe and 3MPA CdTe/CdSe Quantum Dots at PH 12 in Different Periods of Time

This research aims to study the optical characteristics of semiconductor quantum dots (QDs) composed of CdTe and CdTe/CdSe core-shell structures. It utilizes the refluxed method to synthesize these nanoscale particles and aims to comprehend the growth process by monitoring their optical properties over varied periods of time and pH 12. Specifically, the optical evolution of these QDs is evaluated using photoluminescence (PL) and ultraviolet (UV) spectroscopy. For CdTe QDs, a consistent absorbance and peak intensity increase were observed across the spectrum over time. Conversely, CdTe/CdSe QDs displayed distinctive absorbance and peak intensity variations. These disparities might stem from irregularities in forming selenium (Se) layers around CdTe QDs during growth stages, which could potentially induce quenching in the emission spectrum. The optical examinations unveiled a discernible redshift towards higher wavelength values as the reaction progressed. This spectral shift was coupled with an enlargement in QDs size and a decrease in the energy gap. Using PL and UV analysis techniques enabled a comprehensive study of the optical attributes of the CdTe and CdTe/CdSe QD systems. Our findings underscored the influence of growth conditions and shell materials on the optical properties of QDs. The observed changes in absorbance, peak intensity, wavelength values, QDs size, and energy gap with increasing reaction time provided valuable insights into the growth dynamics of these QD structures.

Efficacy and Prognostic Factors of Nonmyeloablative Allogeneic Hematopoietic Stem Cell Transplantation for Patients with Acute Myeloid Leukemia

To retrospectively analyze the efficacy and complications of our institution's modified nonmyeloablative allogeneic hematopoietic stem cell transplantation (NST) in treating intermediate-risk acute myeloid leukemia (AML) - first complete remission (CR1) and prognostic factors. Clinical data of 50 intermediate-risk AML-CR1 patients who underwent matched related NST at the Fifth Medical Center of Chinese People's Liberation Army General Hospital from August 2004 to April 2021 were collected, the hematopoietic recovery, donor engraftment and complications were observed, and overall survival (OS) rate, leukemia-free survival (LFS) rate, treatment-related mortality (TRM), and cumulative relapse rate were calculated. Statistical analysis of factors affecting prognosis was also preformed. The median times for neutrophil and platelet recovery after transplantation were 10 (6-16) and 13 (6-33) days, respectively. One month after transplantation, 22 patients (44%) achieved full donor chimerism (FDC), and 22 patients (44%) achieved mixed chimerism (MC), among whom 18 cases gradually transited to FDC during 1-11 months, 4 cases maintained MC status. The overall incidence of acute graft-versus-host disease (aGVHD) was 36%, with a rate of 18% for grade II-IV aGVHD and a median onset time of 45 (20-70) days after transplantation. The overall incidence of chronic GVHD (cGVHD) was 34%, with 20% and 14% of patients having limited or extensive cGVHD, respectively. The incidence rates of infections, interstitial pneumonia, and hemorrhagic cystitis were 30%, 10%, and 16%, respectively. The 5-year OS rate, LFS rate, TRM, and cumulative relapse rate were 68%, 64%, 16%, and 20%, respectively. The increase of the number of CD34+ cells infused had shortened the recovery time for neutrophils and platelets (r =0.563, r =0.350). The number of CD34+ cells infused significantly influenced the occurrence of extensive cGVHD (OR =1.36, 95%CI : 1.06-1.84, P =0.024). Modified NST is effective in treating intermediate-risk AML-CR1 patients, however, further expansion of sample size is needed to study prognostic factors.

Photoluminescence, morphology and band gap in Europium-doped ZnS nanoparticles

ABSTRACT Current study aims at synthesising Eu-doped ZnS nanoparticles by sol-gel co-precipitation and then characterising them for their structural, optical, and morphological features. Eu-doped ZnS NPs with doping levels of 1, 3, 5, and 7 at.% were synthesised. The characterisation tools such as XRD, UV-Vis, FTIR, photoluminescence and FESEM were employed to analyse their properties. Zinc blend phase with particle sizes lying in the range of 5.0 to 7.2 nm was confirmed by XRD. Eu-doped ZnS NPs show a band gap decrement at a higher level of doping. Defects induced UV and visible emission peaks were observed in photoluminescence emission spectra. Random-shaped particle-like structures that effectively have a persistent enlargement of size were seen in FESEM. The particle size was seen to monitor the band gap at higher doping levels. Eu doping was found to promote the growth of particle sizes which eventually resulted in decrement of the band gap.

Oil density and viscosity affect emulsion stability and destabilization mechanism

The effects of the oils density and viscosity (DV) on the emulsions kinetic stability and destabilization mechanisms were revealed from the perspective of droplets movement and emulsions viscosity. With the extension of storage time, severe creaming (20.20 ± 0.73 × 10−4 mm/s), flocculation and coalescence (7.26 ± 0.82 μm−2 min−1) occurred in emulsions prepared with oils of lowest DV; the extent of creaming (15.90 ± 0.68 × 10−4 mm/s) and coalescence (1.36 ± 0.10 μm−2 min−1) were alleviated upon increasing oils DV, accompanied by the occurrence of acute Ostwald ripening (17.03 ± 1.72 μm3 min−1); and only slight creaming (2.26 ± 0.24 × 10−4 mm/s), flocculation, coalescence (0.70 ± 0.09 μm−2 min−1) and Ostwald ripening (6.14 ± 0.42 μm3 min−1) occurred in emulsions prepared with oils of highest DV. This was because oils with higher DV (especially density) restricted the droplets fluidity due to an increased motion friction, leading to a more homogeneous internal network and a higher apparent/macroscopic viscosity (0.31 ± 0.04 Pa·sn and 0.1 nm−2 s) of emulsions. These impeded the Brownian motion and diffusion of droplets to decrease the collision frequency/efficiency and mass transfer respectively, which in turn conduced to retarding droplets size enlargement and stabilizing the emulsions against different destabilization mechanisms.

MLL-AF9-Induced Leukaemia Changes during Ontogenic Stages

MLL-AF9 causes acute lymphoid leukaemia (ALL), acute myeloid leukaemia (AML) and mixed phenotype acute leukaemia (MPAL) in infants and children. Infant leukaemia originates in utero, yet this has never been formerly shown for MLL-AF9-driven disease. Multiple mouse models invariably revealed that MLL-AF9 can induce myelo-monocytic AML, however, they failed to recapitulate the ALL and MPAL phenotypes as presented in paediatric patients. To explore the impact of developmental stage for MLL-AF9 leukaemic burden and phenotypes, we established a new inducible (“Tet-off”), reversible, transplantable mouse model in which expression of the fusion is controlled by the 3' stem cell leukaemia (SCL)-enhancer element. We crossed SCL-tTA mice(1) with inducible MLL-AF9 mice(2) to generate double transgenic SCLtTA/iMLL-AF9 mice. To determine the impact of developmental stage, MLL-AF9 was induced from the conceptional stage for prenatal (E0), in neonates for postnatal (P0) and at 20 weeks for adult (≥20 weeks) cohorts. Following induction for 6-8 weeks in vivo MLL-AF9 expression was measured in haematopoietic stem cells (HSCs, (Lin-cKit+Sca-1+CD150+CD48-), lymphoid-myeloid primed progenitors (HPC-1/LMPPs, LSK CD150-CD48+/Flt3+), and granulocyte-monocyte progenitors (GMPs, Lin-cKit+Sca-1CD34+CD16/32hi). We found that MLL-AF9 expression levels were comparable within each population across the age groups but observed higher MLL-AF9 expression in the GMPs compared to HSCs in the postnatal model, concomitant with a moderate expansion in the GMP clonal size. Upon postnatal MLL-AF9 induction, all the mice developed leukaemia with a short latency of 13 weeks and complete penetrance. In contrast, E0 prenatal- or adult-induction resulted in longer latencies of 32 and 24 weeks respectively. Despite sustained MLL-AF9 expression, not all the mice developed leukaemia in prenatal- and adult-induced cohorts. Postnatal induced mice developed myelo-monocytic (AML-M4), megakaryoblastic (AML-M7), or MPAL, whereas prenatal and adult induction of MLL-AF9 developed AML-M4 exclusively. Regardless of leukaemia phenotypes, all diseased mice displayed thrombocytopenia and splenomegaly, and leucocytosis. Independent from the age at which MLL-AF9 was induced, all AML-M4 mice had clonal expansion of GMPs. Adult-induced AML-M4 mice had an increase in HSCs while postnatal-induced mice had an increase in HPC-1/LMPPs. In postnatal-induced MPAL mice, there was expansion of GMPs and HPC-1/LMPP populations, and postnatal-induced AML-M7 mice had clonal expansions of HSC, HPC-1/LMPP, and MEP populations with no expansion in GMPs. Thus, the age when the MLL-AF9 fusion is induced does not only determine the disease latency and penetrance, but also affects lineage determination, and the amount of leukaemic HSPCs. Leukaemic-GMP (L-GMP) and leukaemic-HSC (L-HSC) have been characterized in adult models as leukaemic stem cells (LSC). Transplantation of bulk and L-GMPs from adult-induced AML-M4 cells were able to cause AML with complete penetrance, in line with published data and validating our model. However, prenatal- and postnatal-induced AML-M4 cells were not transplantable. In all age cohorts, bulk AML cells formed serially plateable large and compact colonies and were reversible. Collectively, LSCs exist within a GMP-like population of adult-induced MLL-AF9 AML, they may however reside in different HSPC populations in prenatal- and postnatal-induced disease. In summary, this novel inducible model recapitulates several features of infant, paediatric and adult MLL-AF9 AML demonstrating that ontogenic stage is a determining factor in differences in disease biology.

Modulation of the Btla-HVEM Axis to Enhance CAR T Cell Immunotherapy Against Cancer

Introduction: The efficacy of adoptive T cell immunotherapies against cancer, such as chimeric antigen receptor (CAR) T cells, is severely blunted by the immunosuppressive tumor microenvironment (TME). We sought to investigate the role of the TME in cancer resistance to effector T cells in order to define actionable targets to enhance CAR T cell immunotherapies. We initially used Hodgkin lymphoma (HL) as an ideal tumor model since it is characterized by a TME that is profoundly infiltrated by immunosuppressive cells, and then we expanded our findings to multiple cancer models. We first sought to identify the dominant interactions of immunosuppressive cellular compartments and effector T cells by analyzing single-cell RNA sequencing data on a total of 26 (4 exploratory + 22 validation) HL patient tumor biopsies. Using the CellPhoneDB algorithm, we inferred that the ligand B- and T-lymphocyte attenuator (BTLA) on effector T cells and the receptor Herpesvirus entry mediator (HVEM, TNFRSF14) on immunosuppressive cells (e.g., regulatory T cells, monocytes) strongly interact in the TME and promote T cell dysfunction ( Fig 1a). Akin to the canonical checkpoint PD1, BTLA recruits two potent tyrosine phosphatases, SHP-1 and SHP-2, to disable early T cell activation. Thus, we rationalized that BTLA expression on T cells might reduce their anti-tumor function. We hypothesized that deleting BTLA in CAR T cells would abolish BTLA-HVEM trans interactions at the immunological synapse and unleash the cytotoxic potential of CAR T cells. Methods and Results: We first generated BTLA KO anti-CD30 CAR T (CART30) cells against HL. To test their function in vivo, 15x10 6 HDLM-2 (CD30+HVEM+ HL) cells were subcutaneously implanted into NSG mice on day 0, and on day 62, 3x10 5 CAR30+ T cells were infused intravenously. Our results demonstrated that BTLA KO significantly enhances the function of anti-CD30 CAR T cells in HVEM+ HL ( Fig 1b), as assessed via tumor size (caliper) and CART30 expansion in the peripheral blood (flow cytometry). Additionally, we generated BTLA KO 4-1BBζ CART19 cells, which showed greater in vivo anti-tumor function in a subcutaneous tumor model of DLBCL (CD19+ HVEM+ OCI-Ly18). Serum collected from OCI-Ly18-bearing NSG mice infused with BTLA KO CART19 was enriched in effector cytokines (e.g., TNF, IFNγ, IL-2) as measured by Luminex. We then extended these findings into HVEM+ solid tumor models. BTLA KO improved tumor control in vitro in short-term killing experiments (Incucyte SX5) for both CAR and TCR T cells, respectively directed against prostate cancer (HER2+ PC-3) and melanoma (GP-100+ DM-6). Importantly, we demonstrated that BTLA KO in primary BALB/c-derived murine CAR T cells enhances tumor control in A20 murine lymphoma. In this model, BTLA KO tumor-infiltrating CAR T cells showed substantially reduced exhaustion in the TME relative to wild-type CAR T cells. Mechanistically, we showed that targeted mutations in BTLA intracellular tyrosine motifs effectively reduce the recruitment of SHP-1/2 and preserve the docking domain of the pro-stimulatory Grb2, thus maintaining high NFAT signaling and increased persistence in vivo. These effects were dependent on the expression of HVEM on tumor cells. Finally, we found that high BTLA RNA expression in tisagenlecleucel infusion products correlates with poor response to treatment in patients with DLBCL and FL (NCT02030834). Conclusion: Our results reveal a critical role of the BTLA-HVEM axis in inhibiting CAR T cell function, and demonstrate that CRISPR-Cas9 deletion of BTLA leads to enhanced anti-tumor efficacy in multiple models of cancer. The key mechanism is the reduction of SHP-1/2 recruitment and the consequent increase in CAR T cell activation. The results of this study will be translated into a first in human clinical trial of BTLA-deficient CAR T cells for relapsed or refractory cancer.