bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in the Surface Elevation of the Laohugou Glacier No. 12 in Western Qilian Mountains
Authors: Liu Yushuo , Qin Dahe , Li Yanzhao , Qin Xiang , Li Zhenlin , Wang Jing , Jin Zizhen , Wang Lihui
Published: February 2022

As the largest valley glacier in the Qilian Mountains, the Laohugou glacier No. 12 (LHG12) has shrunk significantly since 1957. In this study, two topographic maps and a WorldView-2 satellite stereopair image data were used to assess the volume and cumulative mass balance of LHG12 located at the western Qilian Mountains during 1957–2015. During the study period, the LHG12 exhibited changes in two processes: slightly ablation and stability in a brief period during 1957–1989 and strong melting and accelerated ablation during 1989–2015. During 1957–2015, the volume of LHG12 decreased by 0.38 km3, the average thickness decreased by 17.23 m, the cumulative mass balance (MB) was −14.69 ± 3.00 m w. e., and ablation was found glacier-wide. By comparing the previous MB simulation and digital elevation model (DEM) differencing results, it was found that the MB simulation results underestimated the strong melting trend of LHG12 since the 1990s. Temperature rose, especially in autumn and winter, and could cause the ice temperature of LHG12 to increase, and LHG12 may become more sensitive to climate change.
Energy Evolution and Acoustic Emission Characteristics of Uniaxial Compression Failure of Anchored Layered Sandstone
Authors: Yu Yang , Zhao Da-Cheng , Feng Guang-Liang , Geng Da-Xin , Guo Hao-Sen
Published: February 2022

To study the energy evolution and acoustic emission characteristics of layered sandstone under anchorage in the process of deformation and failure, the sandstone samples from Chuxiong Yi Autonomous Prefecture, Yunnan Province were selected for uniaxial compression testing. The energy evolution in the process of sandstone failure and the spatial fractal characteristics of acoustic emission events in the process of deformation and failure were investigated. Research results show that anchoring can make layered sandstone store more energy, the stored energy first increases, then decreases with the increase of bedding angle; the B value of sandstone under anchorage is generally higher than that of unanchored sandstone in the whole deformation and failure process, and the continuous decline in B value can be used to indicate a precursor to instability and failure; under the action of anchoring, the D value of sandstone (its fractal dimension) also increases, then decreases with the increase of bedding angle. The D value changes within [2, 3]. At a given bedding angle, the D value of anchored sandstone is greater than that of unanchored sandstone, the D value of 30° anchored sandstone increased the most (by 12.33%); the maximum D value occurred in 45° anchored sandstone (reaching 2.72) and the spatial distribution of acoustic emission events and damage of sandstone under anchorage is also more uniform; under increasing stress, the number of acoustic emission events is less widely distributed in the early stage and more densely distributed in the later stage. The growth rate of the D value varies across different peak stress ranges, which is more significant under the action of anchorage. The acoustic emission event counts grow evenly and slowly in the space, and the toughness of sandstone is improved to a certain extent under the action of anchorage.
Recent Advances in the Clinical Application of Adrenal Vein Sampling
Authors: Zhong Shan , Zhang Tianyue , He Minzhi , Yu Hanxiao , Liu Zhenjie , Li Zhongyi , Song Xiaoxiao , Xu Xiaohong
Published: February 2022

We reviewed clinical research investigating the applications of adrenal vein sampling (AVS). AVS could be applied not only to primary aldosteronism (PA) but also to other endocrine diseases, such as adrenocorticotropic hormone (ACTH) independent Cushing syndrome (AICS) and hyperandrogenemia (HA). However, the AVS protocol requires improvements to increase its success rate. Using the computed tomography image fusion, coaxial guidewire technique, and fast intraprocedural cortisol testing (CCF) technique could improve the success rate of catheterization in AVS for PA. ACTH loading could be considered in medical centers with a low selectivity of AVS for PA but is not essential in those with mature AVS technology. The continuous infusion method should be recommended for ACTH stimulation in AVS for PA to reduce adverse events. AVS has not been routinely recommended before management decisions in AICS, but several studies verified that AVS was useful in finding out the source of excess cortisol, especially for distinguishing unilateral from bilateral disease. However, it is necessary to reassess the results of AVS in AICS with the use of reference hormones to fully normalize cortisol levels. In addition, it is essential to determine the optimal model that combines AVS results and mass size to guide the selection of surgical plans, including identifying the dominant gland and presenting the option of staged adrenalectomy, to minimize the impact of bilateral resection. For HA, AVS combined with ovarian intravenous sampling to locate excess androgens could be considered when imaging results are equivocal.
Multipotent Mesenchymal Stromal Cells Interact and Support Islet of Langerhans Viability and Function
Authors: Koehler Naomi , Buhler Leo , Egger Bernhard , Gonelle-Gispert Carmen
Published: February 2022

Type 1 diabetes (T1D) is a widespread disease, affecting approximately 41.5 million people worldwide. It is generally treated with exogenous insulin, maintaining physiological blood glucose levels but also leading to long-term therapeutic complications. Pancreatic islet cell transplantation offers a potential alternative treatment to insulin injections. Shortage of human organ donors has raised the interest for porcine islet xenotransplantation. Neonatal porcine islets are highly available, can proliferate and mature in vitro as well as after transplantation in vivo. Despite promising preclinical results, delayed insulin secretion caused by immaturity and immunogenicity of the neonatal porcine islets remains a challenge for their clinical application. Multipotent mesenchymal stromal cells (MSCs) are known to have pro-angiogenic, anti-inflammatory and immunomodulatory effects. The current state of research emphasizes the great potential of co-culture and co-transplantation of islet cells with MSCs. Studies have shown enhanced islet proliferation and maturation, insulin secretion and graft survival, resulting in an improved graft outcome. This review summarizes the immunomodulatory and anti-inflammatory properties of MSC in the context of islet transplantation.
Experimental and Numerical Study on Thermal Performance of a Novel Direct Expansion Air-Source Heat Pump Heating System
Authors: Wang Haobo , Cheng Yuanda , Wang Dawei , Peng Weijin , Ai Zejian , Jia Jie , Duan Pengfei , Yang Jinming
Published: February 2022

In this paper, a novel direct expansion air-source heat pump heating system with a gravity-driven radiator as a heating terminal was developed, for the purpose of reducing the energy consumption and improving the thermal comfort of indoor personnel. The thermal response speed of the system at the start-up stage was experimentally studied firstly. The results showed that the thermal response speed of the developed system was slightly lower than that of a traditional air-source heat pump heating system with a convective heating terminal, but largely higher than that of an air-source heat pump floor radiant heating system. When the outdoor air temperature fluctuated between −5.2℃ and −1.0℃, the average rising ratios of indoor air temperature at the start-up stage for the developed heating system and the traditional system are 10.8℃/h and 13.8℃/h respectively. On this basis, an optimization study was then conducted to further improve the thermal performance of the developed heating system. The numerical results indicated that integrating a cross-flow fan to the gravity-driven radiator is able to promote the thermal performance of the heating system obviously at the start-up stage, because the convection heat transfer of the radiator was highly enhanced. Among the simulation conditions, the time required for the air temperature at head height to reach the designed temperature is shortest when the flow angle of the fan is 0° with an air speed of 0.6 m/s. while setting the flow angle of the fan to −45° with an air speed of 0.4 m/s is recommended to achieve the most thermal comfort for the whole body.
Promoting Energy Efficiency: Barriers, Societal Needs and Policies
Authors: Della Valle Nives , Bertoldi Paolo
Published: February 2022

The Climate Change urgency requires a swift reduction of energy consumption. One way to achieve this is through increased energy efficiency. Over the past decades, the debate on how to encourage energy efficiency has been guided by the physical–technical–economic model, which has a strong focus on technologies and cost savings, and in which human behaviour has been seen as a trivial factor. However, the advent of behavioural economics has started enabling the integration of the human factor also into energy efficiency policy. Still, this integration is only in its infancy. While the perspectives taken by economics and behavioural sciences enable to capture the individual dimension of energy efficiency as a problem of individual choice, the collective and social aspect of energy efficiency is still largely overlooked on the energy policy agenda. With its emphasis on how social structures interpenetrate individual actions and construction of reality, sociology offers an additional important insight that goes beyond the identification of barriers-drivers underlying investment choices. This paper aims to increase policy makers’ awareness of complementary disciplinary resources, on which they can draw to better define and address the problems associated to energy efficiency. Second, it provides a case to develop an interdisciplinary perspective as a basis to develop a more scientifically valid and socially relevant energy efficiency policy advice.
Nonlinear Seepage Mathematical Model of Fractured Tight Stress Sensitive Reservoir and Its Application
Authors: Yin Daiyin , Li Chunguang , Song Shuang , Liu Kai
Published: February 2022

A tight reservoir can only have industrial production value after fracturing. After volume fracturing, there are artificial fracturing zones and fracturing unmodified zones composed of a matrix, microfractures, and main fractures. There are also pressure-sensitive effects and nonlinear seepage in the matrix. There is an unsteady channeling flow between the matrix and microfractures and main fractures and a threshold pressure gradient in the matrix. The fluid flow between the three media and the transfer flow between the media are very complex. Aiming at the problem of poor simulation accuracy because the above characteristics of tight reservoir fracturing development are not considered in the current commercial numerical simulation software Eclipse, CMG, and VIP, the triple medium coupling nonlinear mathematical model of tight reservoir fracturing development matrix/microfractures/main fractures was established in this study, and the discrete mathematical model was constructed using the full implicit numerical solution method. The model considered the effects of threshold fracturing gradient and nonlinear seepage on the reverse side of seepage. In terms of pressure-sensitive effect, the latest generalized pressure-sensitive effect calculation model, considering rock physical parameters established by authors, was adopted. In terms of matrix/fracture transfer flow, the nonlinear matrix/fracture transfer flow model considering threshold pressure gradient and pressure-sensitive effect established by authors was adopted. Finally, a tight reservoir numerical simulation software was developed based on a triple medium coupling nonlinear model. The software was used to simulate the development effect of turbidite tight reservoir in Wang 587 Block of Jiyang depression, Shengli Oilfield. The results showed that the fitting accuracy of the new model was 13.3% higher than that of Eclipse, and the calculation results were more in line with the production practice of tight reservoirs. The new model established an important theoretical basis for scientifically and effectively guiding the development of tight reservoirs.
Container-Based Sanitation Services and Attrition: An Examination of Drivers and Implications
Authors: VanRiper Froggi , Russel Kory C. , Cramer Lori A. , Tillias Daniel , Laporte Jessica , Lloyd Erica , Kramer Sasha
Published: February 2022

Haiti is far from achieving the UN goal of sanitation access for all; 20% of the population has no sanitation access, and less than 0.1% of the country’s excreta is safely managed. Container-Based Sanitation (CBS) may be key to achieving timely and equitable sanitation coverage in Haiti’s cities. CBS can provide immediate sanitation access without preexisting infrastructure, and where permanent infrastructure is impractical. Investor caution and policy barriers, however, presently limit the growth of CBS solutions. Globally, most CBS services are provided by private organizations like EkoLakay, which provides a portable toilet and weekly excreta collection for a monthly fee. While the EkoLakay service is popular, attrition is high. This study examines the relationship between users and the service, and its role in improved sanitation accessibility. For this study, 633 active and former EkoLakay subscribers in Cap Haïtien were interviewed to reveal causes and implications of attrition. Households with active EkoLakay subscriptions are more likely than former subscribers to live in unauthorized informal residences and to lack energy or water infrastructure. A quarter of users unsubscribe voluntarily, after investing in permanent sanitation infrastructure. Over 30% of former users, however, reported unsubscribing due to economic challenges. Many involuntary terminations resulted in households losing access to private improved sanitation or reverting to open defecation, reducing progress toward global sustainability goals. Insights obtained contextualize the relationship between users and CBS services to inform public strategies for mitigating barriers to achieving universal safe sanitation.
Metabolomic Profiling Reveals That 5-Hydroxylysine and 1-Methylnicotinamide Are Metabolic Indicators of Keloid Severity
Authors: Shan Mengjie , Liu Hao , Hao Yan , Song Kexin , Meng Tian , Feng Cheng , Wang Youbin , Huang Yongsheng
Published: February 2022

Background: Keloid is a skin fibroproliferative disease with unknown pathogenesis. Metabolomics provides a new perspective for revealing biomarkers related to metabolites and their metabolic mechanisms.

Method: Metabolomics and transcriptomics were used for data analysis. Quality control of the data was performed to standardize the data. Principal component analysis (PCA), PLS-DA, OPLS-DA, univariate analysis, CIBERSORT, neural network model, and machine learning correlation analysis were used to calculate differential metabolites. The molecular mechanisms of characteristic metabolites and differentially expressed genes were identified through enrichment analysis and topological analysis.

Result: Compared with normal tissue, lipids have a tendency to decrease in keloids, while peptides have a tendency to increase in keloids. Significantly different metabolites between the two groups were identified by random forest analysis, including 1-methylnicotinamide, 4-hydroxyproline, 5-hydroxylysine, and l-prolinamide. The metabolic pathways which play important roles in the pathogenesis of keloids included arachidonic acid metabolism and d-arginine and d-ornithine metabolism. Metabolomic profiling reveals that 5-hydroxylysine and 1-methylnicotinamide are metabolic indicators of keloid severity. The high-risk early warning index for 5-hydroxylysine is 4 × 108-6.3×108 (p = 0.0008), and the high-risk predictive index for 1-methylnicotinamide is 0.95 × 107-1.6×107 (p = 0.0022).

Conclusion: This study was the first to reveal the metabolome profile and transcriptome of keloids. Differential metabolites and metabolic pathways were calculated by machine learning. Metabolomic profiling reveals that 5-hydroxylysine and 1-methylnicotinamide may be metabolic indicators of keloid severity.
Case Report: Two Families With HPDL Related Neurodegeneration
Authors: Micule Ieva , Lace Baiba , Wright Nathan T. , Chrestian Nicolas , Strautmanis Jurgis , Diriks Mikus , Stavusis Janis , Kidere Dita , Kleina Elfa , Zdanovica Anna , Laflamme Nataly , Rioux Nadie , Setty Samarth Thonta , Pajusalu Sander , Droit Arnaud , Lek Monkol , Rivest Serge , Inashkina Inna
Published: February 2022

There are recent reports of associations of variants in the HPDL gene with a hereditary neurological disease that presents with a wide spectrum of clinical severity, ranging from severe neonatal encephalopathy with no psychomotor development to adolescent-onset uncomplicated spastic paraplegia. Here, we report two probands from unrelated families presenting with severe and intermediate variations of the clinical course. A homozygous variant in the HPDL gene was detected in each proband; however, there was no known parental consanguinity. We also highlight reductions in citrate synthase and mitochondrial complex I activity detected in both probands in different tissues, reflecting the previously proposed mitochondrial nature of disease pathogenesis associated with HPDL mutations. Further, we speculate on the functional consequences of the detected variants, although the function and substrate of the HPDL enzyme are currently unknown.
Genetic Drift and Purifying Selection Shaped Mitochondrial Genome Variation in the High Royal Jelly-Producing Honeybee Strain (Apis mellifera ligustica)
Authors: Ma Chuan , Hu Ruoyang , Costa Cecilia , Li Jianke
Published: February 2022

Mitochondrial genomes (mitogenomes) are involved in cellular energy metabolism and have been shown to undergo adaptive evolution in organisms with increased energy-consuming activities. The genetically selected high royal jelly-producing bees (RJBs, Apis mellifera ligustica) in China can produce 10 times more royal jelly, a highly nutritional and functional food, relative to unselected Italian bees (ITBs). To test for potential adaptive evolution of RJB mitochondrial genes, we sequenced mitogenomes from 100 RJBs and 30 ITBs. Haplotype network and phylogenetic analysis indicate that RJBs and ITBs are not reciprocally monophyletic but mainly divided into the RJB- and ITB-dominant sublineages. The RJB-dominant sublineage proportion is 6-fold higher in RJBs (84/100) than in ITBs (4/30), which is mainly attributable to genetic drift rather than positive selection. The RJB-dominant sublineage exhibits a low genetic diversity due to purifying selection. Moreover, mitogenome abundance is not significantly different between RJBs and ITBs, thereby rejecting the association between mitogenome copy number and royal jelly-producing performance. Our findings demonstrate low genetic diversity levels of RJB mitogenomes and reveal genetic drift and purifying selection as potential forces driving RJB mitogenome evolution.
P2X7 Receptor Triggers Lysosomal Leakage Through Calcium Mobilization in a Mechanism Dependent on Pannexin-1 Hemichannels
Authors: Santos Stephanie Alexia Cristina Silva , Persechini Pedro Muanis , Henriques-Santos Bianca Monteiro , Bello-Santos Victória Gabriela , Castro Newton G. , Costa de Sousa Júlia , Genta Fernando Ariel , Santiago Marcelo Felippe , Coutinho-Silva Robson , Savio Luiz Eduardo Baggio , Kurtenbach Eleonora
Published: February 2022

The P2X7 receptor is a critical purinergic receptor in immune cells. Its activation was associated with cathepsin release into macrophage cytosol, suggesting its involvement in lysosomal membrane permeabilization (LMP) and leakage. Nevertheless, the mechanisms by which P2X7 receptor activation induces LMP and leakage are unclear. This study investigated cellular mechanisms associated with endosomal and lysosomal leakage triggered by P2X7 receptor activation. We found that ATP at 500 μM and 5 mM (but not 50 μM) induced LMP in non-stimulated peritoneal macrophages. This effect was not observed in P2X7-deficient or A740003-pretreated macrophages. We found that the P2X7 receptor and pannexin-1 channels mediate calcium influx that might be important for activating specific ion channels (TRPM2 and two-pore channels) on the membranes of late endosomes and lysosomes leading to LMP leakage and consequent cathepsin release. These findings suggest the critical role of the P2X7 receptor in inflammatory and infectious diseasesvialysosomal dysfunction.
Mass Cytometry Study on Hepatic Fibrosis and Its Drug-Induced Recovery Using Mouse Peripheral Blood Mononuclear Cells
Authors: Bae Jiwon , Kim Ji Eun , Perumalsamy Haribalan , Park Sehee , Kim Yun , Jun Dae Won , Yoon Tae Hyun
Published: February 2022

The number of patients with liver diseases has increased significantly with the progress of global industrialization. Hepatic fibrosis, one of the most common liver diseases diagnosed in many developed countries, occurs in response to chronic liver injury and is primarily driven by the development of inflammation. Earlier immunological studies have been focused on the importance of the innate immune response in the pathophysiology of steatohepatitis and fibrosis, but recently, it has also been reported that adaptive immunity, particularly B cells, plays an essential role in hepatic inflammation and fibrosis. However, despite recent data showing the importance of adaptive immunity, relatively little is known about the role of B cells in the pathogenesis of steatohepatitis fibrosis. In this study, a single-cell-based, high-dimensional mass cytometric investigation of the peripheral blood mononuclear cells collected from mice belonging to three groups [normal chow (NC), thioacetamide (TAA), and 11beta-HSD inhibitor drug] was conducted to further understand the pathogenesis of liver fibrosis through reliable noninvasive biomarkers. Firstly, major immune cell types and their population changes were qualitatively analyzed using UMAP dimensionality reduction and two-dimensional visualization technique combined with a conventional manual gating strategy. The population of B cells displayed a twofold increase in the TAA group compared to that in the NC group, which was recovered slightly after treatment with the 11beta-HSD inhibitor drug. In contrast, the populations of NK cells, effector CD4+T cells, and memory CD8+T cells were significantly reduced in the TAA group compared with those in the NC group. Further identification and quantification of the major immune cell types and their subsets were conducted based on automated clustering approaches [PhenoGraph (PG) and FlowSOM]. The B-cell subset corresponding to PhenoGraph cluster PG#2 (CD62LhighCD44highLy6chighB cells) and PG#3 (CD62LhighCD44highLy6clowB cell) appears to play a major role in both the development of hepatic fibrosis and recoveryviatreatment, whereas PG#1 (CD62LlowCD44highLy6clowB cell) seems to play a dominant role in the development of hepatic fibrosis. These findings provide insights into the roles of cellular subsets of B cells during the progression of, and recovery from, hepatic fibrosis.
Catatonic Schizophrenia Associated With Cerebrospinal GAD65 Autoantibodies: Case Report and Literature Review
Authors: Hansen Niels , Bartels Claudia , Teegen Bianca , Wiltfang Jens , Malchow Berend
Published: February 2022

Background

GAD65 autoimmunity is reported to be associated with schizophrenia and bipolar disorder. However, there has been no evidence that glutamic acid decarboxylase 65 (GAD65) autoantibodies in cerebrospinal fluid (CSF) are associated with akinetic catatonia in schizophrenia patients.

Methods

We report the case of a 28-year-old man who underwent diagnostics including brain MRI, neuropsychological testing, and electroencephalography (EEG) as well as a tumor search via CT of the abdomen and thorax, as well as colonoscopy and gastroscopy. For clinical characterization, his patient files were retrospectively examined.

Results

Our patient presented catatonia that responded somewhat to benzodiazepines in combination with previously taken antipsychotics such as risperidone for prediagnosed paranoid schizophrenia. Diagnostics revealed GAD65 autoantibodies in his serum and CSF. MRI revealed no brain lesion, and the tumor search had no malignancy. We diagnosed catatonic schizophrenia. Furthermore, as he had not fully recovered, he was given immunotherapy entailing two cycles of intravenous immunoglobulins. Subsequent neuropsychological testing due to subjective cognitive complaints after immunotherapy revealed no objective cognitive deficits.

Conclusions

We present the novel finding of an association between GAD65 autoantibodies in the serum and CSF with catatonia in a patient suffering from prediagnosed chronic schizophrenia. Due to the presence of CSF GAD65 antibodies and the catatonia factor in prediagnosed schizophrenia, we suspect that his catatonia has an autoimmune origin. Immunotherapy stabilized the catatonia that had initially responded to lorazepam treatment. Further research should be done to characterize patients’ responses to immunotherapy and standard treatment in a large cohort of patients with GAD65 antibody-associated catatonia and schizophrenia.
Armored BCMA CAR T Cells Eliminate Multiple Myeloma and Are Resistant to the Suppressive Effects of TGF-β
Authors: Alabanza Leah M. , Xiong Ying , Vu Bang , Webster Brian , Wu Darong , Hu Peirong , Zhu Zhongyu , Dropulic Boro , Dash Pradyot , Schneider Dina
Published: February 2022

CAR T-cell therapies targeting the B-cell maturation antigen eliminate tumors in relapsed/refractory multiple myeloma patients, however durable remissions remain difficult to attain. Transforming growth factor beta (TGF-β) is a multifunctional cytokine abundantly expressed in the multiple myeloma bone marrow niche, where it promotes an immunosuppressive tumor microenvironment. We hypothesized that BCMA CAR T-cells armored to resist the suppressive effects of TGF-β will provide an advantage in treating multiple myeloma. The armored B2ARM CAR T cells, co-expressing BCMA targeting CAR with TGF-β dominant-negative receptor II, were generated by lentiviral transduction of primary human CD4+ and CD8+ T cells. The B2ARM CAR T cells eliminated MM.1S multiple myeloma targets in long-term cytotoxicity assays, even under TGF-β-high conditions, whereas cytotoxic function of the non-armored B2 CAR -T cells was inhibited by TGF-β. Concordantly, after long-term exposure to targets in the presence of TGF-β, the B2ARM CAR T cells were enriched for Granzyme B, CD107a, Ki67 and polyfunctional cells T-cells (double or triple-positive for IFN-γ, IL-2 and/or TNF-α), as determined by flow cytometry. In addition, the B2ARM CAR T-cells, but not the conventional B2 CAR T-cells, resisted the TGF-β-mediated suppression of activation (CD25), exhaustion (PD-1, LAG3), and differentiation to T effectors (CD45RA+ CD45RO-CD62L-). In NSG mice bearing RPMI-8226 tumors overexpressing TGF-β, the B2ARM CAR mediated 100% tumor rejection and survival, superior infiltration of tumors on day 7 post CAR T treatment (%CD3+CAR+), and greater expression of IFN-γ, TNF-α, Ki67, Granzyme B, and PD-1, as compared to tumor-infiltrating non-armored B2 CAR T-cells. In NSG RPMI-8226 xenograft model in which tumors were additionally supplemented with TGF-β injections on days -1 through 11 of CAR T treatment, the B2ARM CAR T cells rejected tumors faster than the non-armored B2 CARs, and showed greater numbers of CD3+ and CD3+CAR+, central memory (CD45RO+CD62L+) and effector memory (CD45RO+CD62L-) T cells in the peripheral blood 18 days after treatment. In summary, the armored B2ARM CAR T cells mediate superior persistence, proliferation, multi-functionality, effector differentiation and anti-tumor function in pre-clinical models of multiple myeloma, while abrogating TGF-β-mediated suppression.
Engineering of an Avidity-Optimized CD19-Specific Parallel Chimeric Antigen Receptor That Delivers Dual CD28 and 4-1BB Co-Stimulation
Authors: Halim Leena , Das Kushal K. , Larcombe-Young Daniel , Ajina Adam , Candelli Andrea , Benjamin Reuben , Dillon Richard , Davies David M. , Maher John
Published: February 2022

Co-stimulation is critical to the function of chimeric antigen receptor (CAR) T-cells. Previously, we demonstrated that dual co-stimulation can be effectively harnessed by a parallel (p)CAR architecture in which a CD28-containing second generation CAR is co-expressed with a 4-1BB containing chimeric co-stimulatory receptor (CCR). When compared to linear CARs, pCAR-engineered T-cells elicit superior anti-tumor activity in a range of pre-clinical models. Since CD19 is the best validated clinical target for cellular immunotherapy, we evaluated a panel of CD19-specific CAR and pCAR T-cells in this study. First, we generated a panel of single chain antibody fragments (scFvs) by alanine scanning mutagenesis of the CD19-specific FMC63 scFv (VHdomain) and these were incorporated into second generation CD28+CD3ζ CARs. The resulting panel of CAR T-cells demonstrated a broad range of CD19 binding ability and avidity for CD19-expressing tumor cells. Each scFv-modified CAR was then converted into a pCAR by co-expression of an FMC63 scFv-targeted CCR with a 4-1BB endodomain. When compared to second generation CARs that contained an unmodified or mutated FMC63 scFv, each pCAR demonstrated a significant enhancement of tumor re-stimulation potential and IL-2 release, reduced exhaustion marker expression and enhanced therapeutic efficacy in mice with established Nalm-6 leukemic xenografts. These data reinforce the evidence that the pCAR platform delivers enhanced anti-tumor activity through effective provision of dual co-stimulation. Greatest anti-tumor activity was noted for intermediate avidity CAR T-cells and derived pCARs, raising the possibility that effector to target cell avidity is an important determinant of efficacy.
Interleukin-10 Is a Promising Marker for Immune-Related Adverse Events in Patients With Non-Small Cell Lung Cancer Receiving Immunotherapy
Authors: Wang Haowei , Zhou Fei , Zhao Chao , Cheng Lei , Zhou Caicun , Qiao Meng , Li Xuefei , Chen Xiaoxia
Published: February 2022

Background

Immune checkpoint inhibitors (ICIs) brought about a major paradigm shift in non-small cell lung cancer (NSCLC) treatment. However, the use of ICIs is related to an unforeseeable pattern of immune-related adverse events (irAEs). Hence, more precise biomarkers are needed to predict the incidence of irAEs to prevent overtreatment of ICIs and decrease occurrences of irAEs. This study was designed to identify capable clinical features and plasma inflammatory factors for predicting irAEs.

Methods

A total of 67 patients who received ICI monotherapy or ICI-based combination therapy were retrospectively identified. Clinical characteristics and plasma inflammatory cytokines were collected and analyzed to screen potential biological markers associated with irAEs. The chi-square test, Fisher’s test, and the Mann–Whitney U test were performed for the primary analysis. The optimal cutoff value was determined by a receiver operating characteristic (ROC) curve. Univariate and multivariate logistic regression models were used to identify risk factors of irAEs. Univariate and multivariate Cox proportional hazards were also performed.

Results

Out of 67 patients, 40 (59.7%) experienced irAEs, and 7 (10.4%) experienced severe adverse events (grade ≥ 3). Among these analyzed immune profile biomarkers, only interleukin−10 (IL-10) was related to the risk of irAEs. A high baseline IL−10 plasma level (odds ratio (OR) = 5.318, 95% CI 1.174–24.081, p = 0.030) was found to be a tremendous and independent risk factor for the development of irAEs. Also, for the dynamic analysis, upregulation of IL-10 after one cycle of ICI treatment was positively related to the occurrence of irAEs (OR = 5.712, 95% CI 1.088–29.993, p = 0.039). When pneumonitis, the most common irAEs, was analyzed, only baseline high-expression IL-10 was accompanied with the incidence of pneumonitis (OR = 9.969, 95% CI 1.144–86.843, p = 0.037).

Conclusion

Baseline and dynamic IL-10 plasma levels are tremendously and independently related to higher risk in the development of irAEs and could be utilized for medical practice to monitor adverse events in patients with ICI treatment.
A Seasonal Undercurrent Along the Northwest Coast of Australia
Authors: Furue Ryo
Published: February 2022

In the North West Shelf region of Australia is a surface current (Holloway Current), which flows southwestward along the shelf break. This paper describes a seasonal undercurrent below the Holloway Current. A 5-day climatology is constructed from the output of an eddy-resolving oceanic general circulation model (OGCM). A seasonal northeastward-flowing undercurrent is found on the upper continental slope during the climatological April–May. This undercurrent reverses during February–March. During its annual cycle, the phase of the undercurrent tends to propagate southwestward and upward. The annual frequency dominates, but the positive and negative phases of the undercurrent are not symmetric in the yearly cycle because of the contributions from the semi-annual and 1/3-annual components. We propose a hypothesis that this undercurrent is a beam of coastal trapped wave (CTW). As an initial attempt to assess the plausibility of this hypothesis, we construct an idealized linear coastal-trapped wave (CTW) solution driven by an idealized harmonic meridional winds at the annual frequency. The solution takes the form of a beam originating from the forcing region on the continental shelf and propagating offshore and southward. When it emerges on to the continental slope, it takes the form of an undercurrent. This idealized solution shares several properties with the undercurrent in the OGCM despite several discrepancies.
Integration Bioprocess of B-Phycoerythrin and Exopolysaccharides Production From Photosynthetic Microalga Porphyridium cruentum
Authors: Yin Hao-Chan , Sui Ji-Kang , Han Tian-Li , Liu Tian-Zhong , Wang Hui
Published: February 2022

Red microalgaPorphyridium cruentumhas great potential for converting CO2into high-value bioactive compounds, such as B-phycoerythrin (B-PE) and extracellular polysaccharides or exopolysaccharides (EPS). This study aimed to establish the integration bioprocess of B-PE and EPS production fromP. cruentum. First, different kinds of growth medium and CO2concentration were assessed indoor in terms of high biomass and B-PE and EPS contents. As follows,P. cruentumcells were outdoor scale-up cultured in 700 L pressurized tubular reactors for 9 days till the biomass reached 0.85 g/L and then separated from supernatantsviacentrifugation. Three different methods were adopted to extract phycobiliproteins, and the highest PE contents were extracted from cells by repeated freeze-thawing treatment along with the optimization of significant variables, and finally, 7.99 mg/L B-PE (16,500 Da) with a purity index of 0.82 was obtained. Moreover, analysis of physicochemical properties of EPS extracted fromP. cruentumshowed that the sulfate content was 14.85% and the uronic acid content was 9.36%.
Microbiome Associated With Gambierdiscus balechii Cultures Under Different Toxicity Conditions
Authors: Wu Zhen , Lee Wai Hin , Liu Zijian , Lin Senjie , Lam Paul K. S.
Published: February 2022

Dinoflagellates, including harmful algal bloom species, are known to co-exist with and rely upon bacteria but how the microbiome changes with the physiologies of the cognate dinoflagellates is poorly understood. Here, we used 16S rRNA gene meta-barcoding to characterize the bacterial community in the cultures of Gambierdiscus balechii, a ciguatoxin-producing benthic dinoflagellate, under different nitrogen (N)-nutrient conditions and at different ciguatoxin-producing growth. The high-throughput sequencing of a total of 12 libraries generated 926,438 reads which were classified into 16 phyla. We observed a shift of the G. balechii-associated microbiome from N-replete to low-N conditions and from the early (low toxin) to the late exponential (high toxin) growth stage. Common across these conditions were species from families Rhodobacteraceae and Flavobacteriaceae. Species abundant in the low-N condition mainly included Planctomyces, Ekhidna, and Lactobacillus. Dominant or highly abundant microbial taxa in the high toxin-producing stage (N-replete, late exponential stage) were Oceanococcus and Marinoscillum. Under this condition, one Rhizobiales bacterium, Oricola, also increased in relative abundance. Our study documents the high diversity and dynamics of the G. balechii-associated microbiome, and identifies condition-specific sub-communities: the core (constitutive) microbiome that stably co-exists with G. balechii, the bacterial lineages that are responsive to N-nutrient variations, and species whose abundances are correlated with toxin content of the dinoflagellate. These findings demonstrate that particular bacterial groups are responsive to N-nutrient or toxicity changes of G. balechii and thus will be useful for further investigations on the associated microbiome’s interactions with benthic dinoflagellates and functions in the course of benthic harmful algae blooms.
Long-Term Persisting SARS-CoV-2 RNA and Pathological Findings: Lessons Learnt From a Series of 35 COVID-19 Autopsies
Authors: Maccio Umberto , Zinkernagel Annelies S. , Schuepbach Reto , Probst-Mueller Elsbeth , Frontzek Karl , Brugger Silvio D. , Hofmaenner Daniel Andrea , Moch Holger , Varga Zsuzsanna
Published: February 2022

Background

Long-term sequelae of coronavirus disease 2019 (COVID-19), including the interaction between persisting viral-RNA and specific tissue involvement, pose a challenging issue. In this study, we addressed the chronological correlation (after first clinical diagnosis and postmortem) between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA and organ involvement.

Methods

The presence of postmortem SARS-CoV-2 RNA from 35 complete COVID-19 autopsies was correlated with the time interval between the first diagnosis of COVID-19 and death and with its relationship to morphologic findings.

Results

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA can be evident up to 40 days after the first diagnosis and can persist to 94 hours after death. Postmortem SARS-CoV-2 RNA was mostly positive in lungs (70%) and trachea (69%), but all investigated organs were positive with variable frequency. Late-stage tissue damage was evident up to 65 days after initial diagnosis in several organs. Positivity for SARS-CoV-2 RNA in pulmonary swabs correlated with diffuse alveolar damage (p = 0.0009). No correlation between positive swabs and other morphologic findings was present. Cerebral (p = 0.0003) and systemic hemorrhages (p = 0.009), cardiac thrombi (p = 0.04), and ischemic events (p = 0.03) were more frequent in the first wave, whereas bacterial pneumonia (p = 0.03) was more prevalent in the second wave. No differences in biometric data, clinical comorbidities, and other autopsy findings were found.

Conclusions

Our data provide evidence not only of long-term postmortem persisting SARS-CoV-2 RNA but also of tissue damage several weeks after the first diagnosis of SARS-CoV-2 infection. Additional conditions, such as concomitant bacterial pulmonary superinfection, lung aspergillosis, thromboembolic phenomena, and hemorrhages can further worsen tissue damage.
Evaluation of the Ocular Surface and Meibomian Gland in Obstructive Sleep Apnea Hypopnea Syndrome
Authors: Liu Shaohua , Li Shisheng , Li Mengmeng , Zeng Shiying , Chen Baihua , Zhang Liwei
Published: February 2022

Purpose

To assess the ocular surface and meibomian gland (MG) of patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and to explore the effects of surgery for OSAHS on the ocular surface and MG.

Methods

Based on the apnea hypopnea index (AHI), 21 patients with mild OSAHS (Group A, 5/h ≤ AHI < 15/h), 20 patients with moderate OSAHS (Group B, 15/h ≤ AHI < 30/h), 62 patients with severe OSAHS (Group C, AHI ≥ 30/h) were examined. The ocular surface and MG were evaluated using Keratograph 5M. In addition, detailed Ophthalmic examination including visual acuity, refraction, slit-lamp examination of the anterior segment, corneal fluorescein staining (CFS), ocular surface disease index (OSDI) scoring, Schirmer I test (SIT) and serum lipid measurement was performed. For OSAHS patients with dry eye syndrome (DES) who underwent uvulopalatopharyngoplasty for improving AHI, the conditions of the ocular surface and MG were compared before surgery and 3 months after surgery. Only the data of the right eyes were analyzed.

Results

There were no significantly different in the OSDI score, tear meniscus height (TMH), or loss ratio of the lower eyelid (LRLE) among these groups. The first non-invasive tear film breakup time (fNIBUT), average non-invasive tear film breakup time (avNIBUT), bulbar redness index (BRI), lipid layer grading (LLG), CFS, plugged orifices and distortion in MG, the loss ratio of upper eyelid (LRUE), and the incidence of DES, floppy eyelid syndrome (FES) and meibomian gland dysfunction (MGD) showed significant differences between Groups A and C (p = 0.015, p = 0.018, p < 0.001, p = 0.022, p = 0.036, p = 0.007, p = 0.019, p = 0.017, p = 0.045, p = 0.013, and p = 0.029, respectively). The SIT in the Group A was significantly higher than in Group B (p = 0.025) and in Group C (p < 0.001). In the correlation analyses, the fNIBUT, avNIBUT, SIT and LLG had negative correlations with the AHI (p = 0.013, p = 0.010, p = 0.003, p < 0.001, and p = 0.006, respectively). The BRI, CFS and LRUE were positively correlated with the AHI (p = 0.006, p = 0.007, and p = 0.046, respectively). Three months after surgery, there were no significant differences in the ocular surface or MG.

Conclusion

Patients with severe OSAHS have poor stability of tear film and are prone to lipid-deficient dry eye as a result of the loss of meibomian gland. By improving the AHI, the ocular surface damage of OSAHS patients cannot be reversed in a short time.
River Biofilms Microbiome and Resistome Responses to Wastewater Treatment Plant Effluents Containing Antibiotics
Authors: Matviichuk Olha , Mondamert Leslie , Geffroy Claude , Gaschet Margaux , Dagot Christophe , Labanowski Jérôme
Published: February 2022

Continuous exposure to low concentrations of antibiotics (sub-minimal inhibitory concentration: sub-MIC) is thought to lead to the development of antimicrobial resistance (AMR) in the environmental microbiota. However, the relationship between antibiotic exposure and resistance selection in environmental bacterial communities is still poorly understood and unproven. Therefore, we measured the concentration of twenty antibiotics, resistome quality, and analyzed the taxonomic composition of microorganisms in river biofilms collected upstream (UPS) and downstream (DWS) (at the point of discharge) from the wastewater treatment plant (WWTP) of Poitiers (France). The results of statistical analysis showed that the antibiotic content, resistome, and microbiome composition in biofilms collected UPS were statistically different from that collected DWS. According to Procrustes analysis, microbial community composition and antibiotics content may be determinants of antibiotic resistance genes (ARGs) composition in samples collected DWS. However, network analysis showed that the occurrence and concentration of antibiotics measured in biofilms did not correlate with the occurrence and abundance of antibiotic resistance genes and mobile genetic elements. In addition, network analysis suggested patterns of co-occurrence between several ARGs and three classes of bacteria/algae: Bacteroidetes incertae sedis, Cyanobacteria/Chloroplast, and Nitrospira, in biofilm collected UPS. The absence of a direct effect of antibiotics on the selection of resistance genes in the collected samples suggests that the emergence of antibiotic resistance is probably not only due to the presence of antibiotics but is a more complex process involving the cumulative effect of the interaction between the bacterial communities (biotic) and the abiotic matrix. Nevertheless, this study confirms that WWTP is an important reservoir of various ARGs, and additional efforts and legislation with clearly defined concentration limits for antibiotics and resistance determinants in WWTP effluents are needed to prevent their spread and persistence in the environment.
Development of Loop-Mediated Isothermal Amplification Rapid Diagnostic Assays for the Detection of Klebsiella pneumoniae and Carbapenemase Genes in Clinical Samples
Authors: Poirier Aurore C. , Kuang Dai , Siedler Bianca S. , Borah Khushboo , Mehat Jai W. , Liu Jialin , Tai Cui , Wang Xiaoli , van Vliet Arnoud H. M. , Ma Wei , Jenkins David R. , Clark John , La Ragione Roberto M. , Qu Jieming , McFadden Johnjoe
Published: February 2022

Klebsiella pneumoniae is an important pathogenic bacterium commonly associated with human healthcare and community-acquired infections. In recent years, K. pneumoniae has become a significant threat to global public and veterinary health, because of its high rates of antimicrobial resistance (AMR). Early diagnosis of K. pneumoniae infection and detection of any associated AMR would help to accelerate directed therapy and reduce the risk of the emergence of multidrug-resistant isolates. In this study, we identified three target genes (yhaI, epsL, and xcpW) common to K. pneumoniae isolates from both China and Europe and designed loop-mediated isothermal amplification (LAMP) assays for the detection of K. pneumoniae in clinical samples. We also designed LAMP assays for the detection of five AMR genes commonly associated with K. pneumoniae. The LAMP assays were validated on a total of 319 type reference strains and clinical isolates of diverse genetic backgrounds, in addition to 40 clinical human sputum samples, and were shown to be reliable, highly specific, and sensitive. For the K. pneumoniae–specific LAMP assay, the calculated sensitivity, specificity, and positive and negative predictive values (comparison with culture and matrix-assisted laser desorption/ionization–time of flight mass spectrometry) were all 100% on clinical isolates and, respectively, of 100%, 91%, and 90%, and 100% when tested on clinical sputum samples, while being significantly faster than the reference methods. For the blaKPC and other carbapenemases’ LAMP assays, the concordance between the LAMP results and the references methods (susceptibility tests) was 100%, on both pure cultures (n = 125) and clinical samples (n = 18). In conclusion, we developed highly sensitive and specific LAMP assays for the clinical identification of K. pneumoniae and detection of carbapenem resistance.