bluuebunny/crossref_metadata_2025 huggingface dataset:
Timely Supplementation of Hydrogels Containing Sulfated or Unsulfated Chondroitin and Hyaluronic Acid Affects Mesenchymal Stromal Cells Commitment Toward Chondrogenic Differentiation
Authors: Alessio Nicola , Stellavato Antonietta , Aprile Domenico , Cimini Donatella , Vassallo Valentina , Di Bernardo Giovanni , Galderisi Umberto , Schiraldi Chiara
Published: April 2021

Mesenchymal stromal cells (MSCs) are currently used for cartilage cell therapy because of their well proven capacity to differentiate in chondrocytes. The advantage of MSC-based therapy is the possibility of producing a high number of chondrocytes for implants. The transplant procedure, however, has some limitations, since MSCs may produce non-functional chondrocytes. This limit has been challenged by cultivating MSC in media with hydrogels containing hyaluronic acid (HA), extractive chondroitin sulfate (CS), or bio-fermentative unsulphated chondroitin (BC) alone or in combination. Nevertheless, a clear study of the effect of glycosaminoglycans (GAGs) on chondrocyte differentiation is still lacking, especially for the newly obtained unsulfated chondroitin of biotechnological origin. Are these GAGs playing a role in the commitment of stem cells to chondrocyte progenitors and in the differentiation of progenitors to mature chondrocytes? Alternatively, do they have a role only in one of these biological processes? We evaluated the role of HA, CS, and – above all – BC in cell commitment and chondrocyte differentiation of MSCs by supplementing these GAGs in different phases of in vitro cultivation. Our data provided evidence that a combination of HA and CS or of HA and BC supplemented during the terminal in vitro differentiation and not during cell commitment of MSCs improved chondrocytes differentiation without the presence of fibrosis (reduced expression of Type I collagen). This result suggests that a careful evaluation of extracellular cues for chondrocyte differentiation is fundamental to obtaining a proper maturation process.
Expressional and Prognostic Value of S100A16 in Pancreatic Cancer Via Integrated Bioinformatics Analyses
Authors: Tu Gangping , Gao Wenzhe , Li Ying , Dian Yating , Xue Bingyang , Niu Li , Yu Xiao , Zhu Hongwei
Published: April 2021

Studies have shown that the calcium-binding protein family S100 may play a role in the development of pancreatic cancer (PC), but the role of S100A16 in PC is still unknown. In this study, Oncomine was first used to detect the expression level and prognosis of S100A16 in PC and other tumors. The results showed that S100A16 was highly expressed in PC tissues compared with a normal pancreas, and the increased expression level may be related to poor prognosis in PC patients. The TCGA and ICGC RNA-seq data of PC patients were downloaded, and the S100A16-related differentially expressed genome (DEGs) was defined by taking the intersection of two gene sets. The GO and KEGG pathways were then analyzed. For clinical analysis, boxplots were depicted for the correlation between clinical characteristics and S100A16 expression. Then Cox regression was applied for exploring the prognostic value of S100A16 for PDAC patients. Based on the Cox regression model, we further estabished a S100A16-related risk score system to strengthen the ability to predict patients' prognosis. After integrating the risk score model and multiple clinicopathological factors, we finally established a nomogram that could predict the survival time of patients. Moreover, Gene set enrichment the effect of S100A16 expression differences on downstream biological processes. At last, using TIMER, ImmuneCellAI and GSEA we analyzed the correlation between S100A16 and pancreatic cancer immune infiltration and predicted the response of patients to checkpoint Blocker (ICB). In summary, S100A16 is involved in the occurrence and development of PC, affecting the prognosis of patients, and may have potential reference values for the immunotherapy of PC.
An ω-3, but Not an ω-6 Polyunsaturated Fatty Acid Decreases Membrane Dipole Potential and Stimulates Endo-Lysosomal Escape of Penetratin
Authors: Zakany Florina , Szabo Mate , Batta Gyula , Kárpáti Levente , Mándity István M. , Fülöp Péter , Varga Zoltan , Panyi Gyorgy , Nagy Peter , Kovacs Tamas
Published: April 2021

Although the largely positive intramembrane dipole potential (DP) may substantially influence the function of transmembrane proteins, its investigation is deeply hampered by the lack of measurement techniques suitable for high-throughput examination of living cells. Here, we describe a novel emission ratiometric flow cytometry method based on F66, a 3-hydroxiflavon derivative, and demonstrate that 6-ketocholestanol, cholesterol and 7-dehydrocholesterol, saturated stearic acid (SA) and ω-6 γ-linolenic acid (GLA) increase, while ω-3 α-linolenic acid (ALA) decreases the DP. These changes do not correlate with alterations in cell viability or membrane fluidity. Pretreatment with ALA counteracts, while SA or GLA enhances cholesterol-induced DP elevations. Furthermore, ALA (but not SA or GLA) increases endo-lysosomal escape of penetratin, a cell-penetrating peptide. In summary, we have developed a novel method to measure DP in large quantities of individual living cells and propose ALA as a physiological DP lowering agent facilitating cytoplasmic entry of penetratin.
Single-Cell Atlas Reveals Fatty Acid Metabolites Regulate the Functional Heterogeneity of Mesenchymal Stem Cells
Authors: Xie Jiayi , Lou Qi , Zeng Yunxin , Liang Yingying , Xie Siyu , Xu Quanhui , Yuan Lisha , Wang Jin , Jiang Linjia , Mou Lisha , Lin Dongjun , Zhao Meng
Published: April 2021

Bone marrow mesenchymal stem cells (MSCs) are widely used clinically due to their versatile roles in multipotency, immunomodulation, and hematopoietic stem cell (HSC) niche function. However, cellular heterogeneity limits MSCs in the consistency and efficacy of their clinical applications. Metabolism regulates stem cell function and fate decision; however, how metabolites regulate the functional heterogeneity of MSCs remains elusive. Here, using single-cell RNA sequencing, we discovered that fatty acid pathways are involved in the regulation of lineage commitment and functional heterogeneity of MSCs. Functional assays showed that a fatty acid metabolite, butyrate, suppressed the self-renewal, adipogenesis, and osteogenesis differentiation potential of MSCs with increased apoptosis. Conversely, butyrate supplement significantly promoted HSC niche factor expression in MSCs, which suggests that butyrate supplement may provide a therapeutic approach to enhance their HSC niche function. Overall, our work demonstrates that metabolites are essential to regulate the functional heterogeneity of MSCs.
Metabolic Syndrome and Autophagy: Focus on HMGB1 Protein
Authors: Frisardi Vincenza , Matrone Carmela , Street Maria Elisabeth
Published: April 2021

Metabolic syndrome (MetS) affects the population worldwide and results from several factors such as genetic background, environment and lifestyle. In recent years, an interplay among autophagy, metabolism, and metabolic disorders has become apparent. Defects in the autophagy machinery are associated with the dysfunction of many tissues/organs regulating metabolism. Metabolic hormones and nutrients regulate, in turn, the autophagy mechanism. Autophagy is a housekeeping stress-induced degradation process that ensures cellular homeostasis. High mobility group box 1 (HMGB1) is a highly conserved nuclear protein with a nuclear and extracellular role that functions as an extracellular signaling molecule under specific conditions. Several studies have shown that HMGB1 is a critical regulator of autophagy. This mini-review focuses on the involvement of HMGB1 protein in the interplay between autophagy and MetS, emphasizing its potential role as a promising biomarker candidate for the early stage of MetS or disease’s therapeutic target.
Back to the Future: From Appendage Development Toward Future Human Hair Follicle Neogenesis
Authors: de Groot Simon C. , Ulrich Magda M. W. , Gho Coen G. , Huisman Margriet A.
Published: April 2021

Hair disorders such as alopecia and hirsutism often impact the social and psychological well-being of an individual. This also holds true for patients with severe burns who have lost their hair follicles (HFs). HFs stimulate proper wound healing and prevent scar formation; thus, HF research can benefit numerous patients. Although hair development and hair disorders are intensively studied, human HF development has not been fully elucidated. Research on human fetal material is often subject to restrictions, and thus development, disease, and wound healing studies remain largely dependent on time-consuming and costly animal studies. Although animal experiments have yielded considerable and useful information, it is increasingly recognized that significant differences exist between animal and human skin and that it is important to obtain meaningful human models. Human disease specific models could therefore play a key role in future therapy. To this end, hair organoids or hair-bearing skin-on-chip created from the patient’s own cells can be used. To create such a complex 3D structure, knowledge of hair genesis, i.e., the early developmental process, is indispensable. Thus, uncovering the mechanisms underlying how HF progenitor cells within human fetal skin form hair buds and subsequently HFs is of interest. Organoid studies have shown that nearly all organs can be recapitulated as mini-organs by mimicking embryonic conditions and utilizing the relevant morphogens and extracellular matrix (ECM) proteins. Therefore, knowledge of the cellular and ECM proteins in the skin of human fetuses is critical to understand the evolution of epithelial tissues, including skin appendages. This review aims to provide an overview of our current understanding of the cellular changes occurring during human skin and HF development. We further discuss the potential implementation of this knowledge in establishing a humanin vitromodel of a full skin substitute containing hair follicles and the subsequent translation to clinical use.
Recent Advances on the Applications of Luminescent Pb2+-Containing Metal–Organic Frameworks in White-Light Emission and Sensing
Authors: Wang Li-Xin , Xiang Jing , Li Chuan-Hua , Leung Chi-Fai , Xiang Jing
Published: April 2021

Luminescent Pb2+-based metal–organic frameworks (MOFs) belong to a new class of multifunctional molecular materials with interesting luminescence properties and potential applications within a single crystalline phase. In this mini review, we present the recent advances that have been achieved in their applications as single-phase white-light emitting materials and chemosensors in the last decade. We focus on the trends in the modification of their structures and luminescence by various bridging ligands, and subsequently their multifunctional applications, which may affect the future development of the field.
Detection and Classification of Multi-Type Cells by Using Confocal Raman Spectroscopy
Authors: Wen Jing , Tang Tianchen , Kanwal Saima , Lu Yongzheng , Tao Chunxian , Zheng Lulu , Zhang Dawei , Gu Zhengqin
Published: April 2021

Tumor cells circulating in the peripheral blood are the prime cause of cancer metastasis and death, thus the identification and discrimination of these rare cells are crucial in the diagnostic of cancer. As a label-free detection method without invasion, Raman spectroscopy has already been indicated as a promising method for cell identification. This study uses a confocal Raman spectrometer with 532 nm laser excitation to obtain the Raman spectrum of living cells from the kidney, liver, lung, skin, and breast. Multivariate statistical methods are applied to classify the Raman spectra of these cells. The results validate that these cells can be distinguished from each other. Among the models built to predict unknown cell types, the quadratic discriminant analysis model had the highest accuracy. The demonstrated analysis model, based on the Raman spectrum of cells, is propitious and has great potential in the field of biomedical for classifying circulating tumor cells in the future.
Adjunctive S100A8/A9 Immunomodulation Hinders Ciprofloxacin Resistance in Pseudomonas aeruginosa in a Murine Biofilm Wound Model
Authors: Laulund Anne S. , Schwartz Franziska , Trøstrup Hannah , Thomsen Kim , Christophersen Lars , Calum Henrik , Ciofu Oana , Høiby Niels , Moser Claus
Published: April 2021

Objective

Pseudomonas aeruginosa is known to contribute to the pathogenesis of chronic wounds by biofilm-establishment with increased tolerance to host response and antibiotics. The neutrophil-factor S100A8/A9 has a promising adjuvant effect when combined with ciprofloxacin, measured by quantitative bacteriology, and increased anti- and lowered pro-inflammatory proteins. We speculated whether a S100A8/A9 supplement could prevent ciprofloxacin resistance in infected wounds.

Method

Full-thickness 2.9cm2-necrosis was inflicted on 32 mice. On day 4, P.aeruginosa in seaweed alginate was injected sub-eschar to mimic a mono-pathogenic biofilm. Mice were randomized to receive ciprofloxacin and S100A8/A9 (n=14), ciprofloxacin (n=12) or saline (n=6). Half of the mice in each group were euthanized day 6 and the remaining day 10 post-infection. Mice were treated until sacrifice. Primary endpoint was the appearance of ciprofloxacin resistant P.aeruginosa. The study was further evaluated by genetic characterization of resistance, means of quantitative bacteriology, wound-size and cytokine-production.

Results

Three mice receiving ciprofloxacin monotherapy developed resistance after 14 days. None of the mice receiving combination therapy changed resistance pattern. Sequencing of fluoroquinolone-resistance determining regions in the ciprofloxacin resistant isolates identified two high-resistant strains mutated in gyrA C248T (MIC>32µg/ml) and a gyr B mutation was found in the sample with low level resistance (MIC=3µg/ml). Bacterial densities in wounds were lower in the dual treated group compared to the placebo group on both termination days.

Conclusion

This study supports the ciprofloxacin augmenting effect and indicates a protective effect in terms of hindered ciprofloxacin resistance of adjuvant S100A8/A9 in P.aeruginosa biofilm infected chronic wounds.
The Modular Circuitry of Apicomplexan Cell Division Plasticity
Authors: Gubbels Marc-Jan , Coppens Isabelle , Zarringhalam Kourosh , Duraisingh Manoj T. , Engelberg Klemens
Published: April 2021

The close-knit group of apicomplexan parasites displays a wide variety of cell division modes, which differ between parasites as well as between different life stages within a single parasite species. The beginning and endpoint of the asexual replication cycles is a ‘zoite’ harboring the defining apical organelles required for host cell invasion. However, the number of zoites produced per division round varies dramatically and can unfold in several different ways. This plasticity of the cell division cycle originates from a combination of hard-wired developmental programs modulated by environmental triggers. Although the environmental triggers and sensors differ between species and developmental stages, widely conserved secondary messengers mediate the signal transduction pathways. These environmental and genetic input integrate in division-mode specific chromosome organization and chromatin modifications that set the stage for each division mode. Cell cycle progression is conveyed by a smorgasbord of positively and negatively acting transcription factors, often acting in concert with epigenetic reader complexes, that can vary dramatically between species as well as division modes. A unique set of cell cycle regulators with spatially distinct localization patterns insert discrete check points which permit individual control and can uncouple general cell cycle progression from nuclear amplification. Clusters of expressed genes are grouped into four functional modules seen in all division modes: 1. mother cytoskeleton disassembly; 2. DNA replication and segregation (D&S); 3. karyokinesis; 4. zoite assembly. A plug-and-play strategy results in the variety of extant division modes. The timing of mother cytoskeleton disassembly is hard-wired at the species level for asexual division modes: it is either the first step, or it is the last step. In the former scenario zoite assembly occurs at the plasma membrane (external budding), and in the latter scenario zoites are assembled in the cytoplasm (internal budding). The number of times each other module is repeated can vary regardless of this first decision, and defines the modes of cell division: schizogony, binary fission, endodyogeny, endopolygeny.
Regulation of Endothelial and Vascular Functions by Carbon Monoxide via Crosstalk With Nitric Oxide
Authors: Choi Yoon Kyung , Kim Young-Myeong
Published: April 2021

Carbon monoxide (CO), generated by heme oxygenase (HO), has been considered a signaling molecule in both the cardiovascular and central nervous systems. The biological function of the HO/CO axis is mostly related to other gaseous molecules, including nitric oxide (NO), which is synthesized by nitric oxide synthase (NOS). Healthy blood vessels are essential for the maintenance of tissue homeostasis and whole-body metabolism; however, decreased or impaired vascular function is a high-risk factor of cardiovascular and neuronal diseases. Accumulating evidence supports that the interplay between CO and NO plays a crucial role in vascular homeostasis and regeneration by improving endothelial function. Moreover, endothelial cells communicate with neighboring cells, such as, smooth muscle cells, immune cells, pericytes, and astrocytes in the periphery and neuronal vascular systems. Endogenous CO could mediate the cell-cell communication and improve the physiological functions of the cardiovascular and neurovascular systems via crosstalk with NO. Thus, a forward, positive feedback circuit between HO/CO and NOS/NO pathways can maintain cardiovascular and neurovascular homeostasis and prevent various human diseases. We discussed the crucial role of CO-NO crosstalk in the cardiovascular and neurovascular systems.
A One-Class-Classifier-Based Negative Data Generation Method for Rapid Earthquake-Induced Landslide Susceptibility Mapping
Authors: Chen Shuai , Miao Zelang , Wu Lixin , Zhang Anshu , Li Qirong , He Yueguang
Published: April 2021

Machine learning with extensively labeled training samples (e.g., positive and negative data) has received much attention in terms of addressing earthquake-induced landslide susceptibility mapping (LSM). However, the extensive amount of labeled training data required by machine learning, particularly the precise negative data (i.e., non-landslide area), cannot be easily and efficiently collected. To address this issue, this study presents a one-class-classifier-based negative data generation method for rapid earthquake-induced LSM. First, an incomplete landslide inventory (i.e., positive data) was produced with the aid of change detection using before-and-after satellite images and the Geographic Information System (GIS). Second, a one-class classifier was utilized to compute the probability of landslide occurrence based on the incomplete landslide inventory followed by the negative data generation from the low landslide susceptibility areas. Third, the positive data as well as the generated negative data (i.e., non-landslide) were compounded to train a traditional binary classifier to produce the final LSM. Experimental results suggest that the proposed method is capable of achieving a result that is comparable to methods using the complete landslide inventory, and it displays good correspondence with recent landslide events, making it a suitable method for rapid earthquake-induced LSM. The findings in this study would be useful in regional disaster planning and risk reduction.
Dominant Role of Sea Level on the Sedimentary Environmental Evolution in the Bohai and Yellow Seas Over the Last 1 Million Years
Authors: Shi Xuefa , Yao Zhengquan , Liu Jianxing , Qiao Shuqing , Liu Yanguang , Li Xiaoyan , Fang Xisheng , Li Chaoxin
Published: April 2021

Transgression and regression deposits from the shallow continental margin provide information on orbital-scale variations in sea level, climate change, and local tectonics. In this study, we conduct a high-resolution chronological and sedimentological analysis of a 125-m core (NHH01) drilled in the southern Yellow Sea. We developed a high-resolution age model at the orbital timescale over the last ∼1 Myr by the astronomical tuning of the sediment grain size. Sedimentological analysis and environmental proxies reveal that the study area is characterized by cyclic alternations of neritic and littoral/fluvial deposits controlled by glacial–interglacial sea-level changes. These results confirm the earlier assumption that sea-level fluctuations play a dominant role in the sedimentary architecture of the southern Yellow Sea. Moreover, only low-frequency sea-level fluctuations (∼100 kyr) were preserved in the NHH01 sequence; however, additional high-frequency (∼40 kyr) sea-level variations were also present in the sediments of the shallower Bohai Sea. Despite the large spatial difference, this finding implies that the sedimentary environment in the eastern marginal seas of China had been influenced by the sea level as a whole over the last 1 Myr. The comparison of the sedimentary environment with other cores from the Bohai Sea, Yellow Sea, and coastal region, as well as records from Indonesia and Japan, reveals that a consistent initiation of large-scale marine transgression occurred at ∼0.8–1 Ma. This phenomenon was likely caused by the long duration of glacio-eustatic high sea-level stands following the Middle Pleistocene transition, which is characterized by a shift in glacial cycles from 40 to 100 kyr.
Predictive Factors for Live Birth in Fresh In Vitro Fertilization/Intracytoplasmic Sperm Injection Treatment in Poor Ovarian Reserve Patients Classified by the POSEIDON Criteria
Authors: Li Fei , Ye Tian , Kong Huijuan , Li Jing , Hu Linli , Jin HaiXia , Guo YiHong , Li Gang
Published: April 2021

The mechanisms underlying poor ovarian response (POR) in assisted reproductive technology remain unclear, there is no consensus on the management of poor responders, the POSEIDON stratification classifies infertility patients into “expected” or “unexpected” groups to provide a more nuanced picture of POR, but few researchers have discussed the independent predictive factors (smoothed plots and the threshold effect) for live birth in POR patients classified by the new criteria. We conducted a retrospective cohort study using clinical data from 6,580 POR patients classified by the POSEIDON criteria in the First Affiliated Hospital of Zhengzhou University, and explored the live birth based on the results before and after the threshold inflection point of each independent influencing factor. Among 6,580 poor ovarian reserve patients classified by the POSEIDON criteria, 1,549 (23.54%) had live births, and 5,031 (76.46%) did not have live births. Multivariate logistic regression analysis showed that female age (OR 0.901; 95% CI 0.887~0.916; P < 0.001), body mass index (OR 0.963; 95% CI 0.951~0.982; P < 0.001), antral follicle counting (OR 1.049; 95% CI 1.009~1.042; P < 0.001) and controlled ovarian hyperstimulation protocol were independent factors predicting live birth in patients with POR. The threshold effect analysis found that the inflection point of female age was 34 years old, and when age was > 34 years old, the probability of live birth in POR patients dropped sharply (OR 0.7; 95% CI 0.7~0.8; P < 0.001). The inflection point of BMI was 23.4 kg/m2, and BMI had a negative correlation with live birth (OR 0.963; 95% CI 0.951~0.982; P < 0.001). The threshold inflection point of AFC was 8n. Female age, BMI, AFC and COH protocol were independent predictive factors associated with live birth in POR patients classified by the POSEIDON criteria. The smooth curve fit and threshold effect analyses provide clinical management strategies for these patients. In addition, the early-follicular-phase long-acting GnRH-agonist long protocol seems to have a higher live birth rates than other protocols. It is worth highlighting that BMI should be considered as well in the POSEIDON criteria.
Polyether-Based Supramolecular Electrolytes With Two-Dimensional Boroxine Skeleton
Authors: Yoshizawa-Fujita Masahiro , Horiuchi Shunsuke , Uemiya Tamao , Ishii Jun , Takeoka Yuko , Rikukawa Masahiro
Published: April 2021

Solid polymer electrolytes mainly based on polyethers have been actively investigated for over 40 years to develop safe, light, and flexible rechargeable batteries. Here, we report novel supramolecular electrolytes (SMEs) composed of polyether derivatives and a two-dimensional boroxine skeleton synthesized by the dehydration condensation of 1,4-benzenediboronic acid in the presence of a polyether with amines on both chain ends. The formation of SMEs based on polyether derivatives and boroxine skeleton was confirmed by Fourier transform infrared (FT-IR) spectroscopy, X-ray photoelectron spectroscopy (XPS), powder X-ray diffraction (PXRD), and thermogravimetric (TG) analysis. Linear sweep voltammetry (LSV) and cyclic voltammetry (CV) were performed to evaluate the electrochemical stability and lithium conductive properties of SMEs with given amounts of lithium bis(trifluoromethylsulfonyl)amide (LiTFSA). The ionic conductivity of SME/LiTFSA composites increased with increasing lithium-salt concentration and reached a maximum value at a higher concentration than those of simple polyether systems. The lithium-ion transference number (tLi+) of SME/LiTFSA was higher than those of polyether electrolytes. This tendency is unusual for a polyether matrix. SME/LiTFSA composite electrolytes exhibited a stable lithium plating/striping process even after 100 cycles. The current density increased with an increasing number of cycles. The combination of ion conductive polymers and a two-dimensional boroxine skeleton will be an interesting concept for developing solid electrolytes with good electrochemical properties.
Heterozygous Single-Nucleotide Polymorphism Genotypes at Heat Shock Protein 70 Gene Potentially Influence Thermo-Tolerance Among Four Zebu Breeds of Nigeria
Authors: Onasanya Gbolabo Olaitan , Msalya George Mutani , Thiruvenkadan Aranganoor Kannan , Sreekumar Chirukandoth , Tirumurugaan Gopalan Krishnaswamy , Fafiolu Adeboye O. , Adeleke Matthew A. , Yakubu Abdulmojeed , Ikeobi Christian Obiora Ndubuisi , Okpeku Moses
Published: April 2021

Genetic variants at heat shock protein 70 gene and their influence on heat stress (HS) tolerance were studied among selected Nigeria zebu, namely, 25 White Fulani (WF), 21 Sokoto Gudali (SG), 21 Red Bororo (RB), and 23 Ambala (AM). Detection of single nucleotide polymorphism (SNP) followed by determination of genotype and genotypic frequency was made among the selected breeds. The heat tolerance coefficient (HTC) was determined from thermo-related parameters including body temperature, rectal temperature, and respiratory rate. Thermo-Tolerance was evaluated through the SNP–thermo-parameter relationship. Statistical analyses were done using the GLM procedure in SAS. A quantitative real-time/high-resolution melting-based assay detected twelve genetic variants. Five of these were common and shared across all breeds of cattle. Of the remaining seven variants, three were specifically identified in AM, two in SG, and two in RB. Also, SNPs were evaluated and four unique SNPs (C151T, C146T, G90A, and C219A) were identified. Heterozygous animals had lower HTC suggesting their potential to withstand HS than homozygous counterparts. The WF and RB animals had significantly lower values for all parameters (BT, RT, RR, and HTC) compared to AM and SG breeds. Thermo-related parameters were significantly different (P< 0.001), and it is recommended that screening of SNPs in zebu is needed to enable selection for improved thermo-tolerance.
Phenotypic and Epigenetic Adaptations of Cord Blood CD4+ T Cells to Maternal Obesity
Authors: Sureshchandra Suhas , Mendoza Norma , Jankeel Allen , Wilson Randall M. , Marshall Nicole E. , Messaoudi Ilhem
Published: April 2021

Pregravid obesity has been shown to disrupt the development of the offspring’s immune system and increase susceptibility to infection. While the mechanisms underlying the impact of maternal obesity on fetal myeloid cells are emerging, the consequences for T cells remain poorly defined. In this study, we collected umbilical cord blood samples from infants born to lean mothers and mothers with obesity and profiled CD4 T cells using flow cytometry and single cell RNA sequencing at resting and following ex vivo polyclonal stimulation. We report that maternal obesity is associated with higher frequencies of memory CD4 T cells suggestive of in vivo activation. Moreover, single cell RNA sequencing revealed expansion of an activated subset of memory T cells with maternal obesity. However, ex vivo stimulation of purified CD4 T cells resulted in poor cytokine responses, suggesting functional defects. These phenotypic and functional aberrations correlated with methylation and chromatin accessibility changes in loci associated with lymphocyte activation and T cell receptor signaling, suggesting a possible link between maternal obesogenic environment and fetal immune reprogramming. These observations offer a potential explanation for the increased susceptibility to microbial infection in babies born to mothers with obesity.
The Traf2DNxBCL2-tg Mouse Model of Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma Recapitulates the Biased IGHV Gene Usage, Stereotypy, and Antigen-Specific HCDR3 Selection of Its Human Counterpart
Authors: Perez-Chacon Gema , Zapata Juan M.
Published: April 2021

Chronic lymphocytic leukemia (CLL)/Small lymphocytic lymphoma (SLL) is a heterogeneous disease consisting of at least two separate subtypes, based on the mutation status of the immunoglobulin heavy chain variable gene (IGHV) sequence. Exposure to antigens seems to play a role in malignant transformation and in the selection and expansion of more aggressive CLL clones. Furthermore, a biased usage of particular IGHV gene subgroups and the existence of stereotyped B-cell receptors (BCRs) are distinctive characteristics of human CLL. We have previously described thatTraf2DN/BCL2double-transgenic (tg,+/+) mice develop CLL/SLL with high incidence with aging. In this model, TNF-Receptor Associated Factor (TRAF)-2 deficiency cooperates with B cell lymphoma (BCL)-2 in promoting CLL/SLL in mice by specifically enforcing marginal zone (MZ) B cell differentiation and rendering B cells independent of BAFF for survival. In this report, we have performed the sequencing of the IGHV-D-J rearrangements of B cell clones from theTraf2DN/BCL2-tg+/+mice with CLL/SLL. The results indicate that these mice develop oligoclonal and monoclonal B cell expansions. Allotransplantation of the oligoclonal populations into immunodeficient mice resulted in the preferential expansion of one of the parental clones. The analysis of the IGHV sequences indicated that 15% were mutated (M) and 85% unmutated (UM). Furthermore, while theTraf2DN/BCL2-tg-/-(wild-type),-/+(BCL2single-tg) and+/-(Traf2DNDN single-tg) littermates showed the expression of various IGHV gene subgroups, the CLL/SLL expanded clones from theTraf2DN/BCL2-tg+/+(double-transgenic) mice showed a more restricted IGHV gene subgroup usage and an overrepresentation of particular IGHV genes. In addition, the HCDR3-encoded protein sequence indicates the existence of stereotyped immunoglobulin (Ig) in the BCRs and strong similarities with BCR recognizing autoantigens and pathogen-associated antigens. Altogether, these results highlight the remarkable similarities between the CLL/SLL developed by theTraf2DN/BCL2-tg+/+mice and its human counterpart.
Subversion of Host Innate Immunity by Rickettsia australis via a Modified Autophagic Response in Macrophages
Authors: Bechelli Jeremy , Rumfield Claire S. , Walker David H. , Widen Steven , Khanipov Kamil , Fang Rong
Published: April 2021

We recently reported that thein vitroandin vivosurvivals ofRickettsia australisareAtg5-dependent, in association with an inhibited level of anti-rickettsial cytokine, IL-1β. In the present study, we sought to investigate howR. australisinteracts with host innate immunityviaanAtg5-dependent autophagic response. We found that the serum levels of IFN-γ and G-CSF inR. australis-infectedAtg5flox/floxLyz-Cremice were significantly less compared toAtg5flox/floxmice, accompanied by significantly lower rickettsial loads in tissues with inflammatory cellular infiltrations including neutrophils.R. australisinfection differentially regulated a significant number of genes in bone marrow-derived macrophages (BMMs) in anAtg5-depdent fashion as determined by RNA sequencing and Ingenuity Pathway Analysis, including genes in the molecular networks of IL-1 family cytokines and PI3K-Akt-mTOR. The secretion levels of inflammatory cytokines, such as IL-1α, IL-18, TNF-α, and IL-6, byR. australis-infectedAtg5flox/floxLyz-CreBMMs were significantly greater compared to infectedAtg5flox/floxBMMs. Interestingly, R. australissignificantly increased the levels of phosphorylated mTOR and P70S6K at a time when the autophagic response is induced. Rapamycin treatment nearly abolished the phosphorylated mTOR and P70S6K but did not promote significant autophagic flux duringR. australisinfection. These results highlight thatR. australismodulates anAtg5-dependent autophagic response, which is not sensitive to regulation by mTORC1 signaling in macrophages. Overall, we demonstrate thatR. australiscounteracts host innate immunity including IL-1β-dependent inflammatory response to support the bacterial survivalviaan mTORC1-resistant autophagic response in macrophages.
Vasculitis and Neutrophil Extracellular Traps in Lungs of Golden Syrian Hamsters With SARS-CoV-2
Authors: Becker Kathrin , Beythien Georg , de Buhr Nicole , Stanelle-Bertram Stephanie , Tuku Berfin , Kouassi Nancy Mounogou , Beck Sebastian , Zickler Martin , Allnoch Lisa , Gabriel Gülsah , von Köckritz-Blickwede Maren , Baumgärtner Wolfgang
Published: April 2021

Neutrophil extracellular traps (NETs) have been identified as one pathogenetic trigger in severe COVID-19 cases and therefore well-described animal models to understand the influence of NETs in COVID-19 pathogenesis are needed. SARS-CoV-2 infection causes infection and interstitial pneumonia of varying severity in humans and COVID-19 models. Pulmonary as well as peripheral vascular lesions represent a severe, sometimes fatal, disease complication of unknown pathogenesis in COVID-19 patients. Furthermore, neutrophil extracellular traps (NETs), which are known to contribute to vessel inflammation or endothelial damage, have also been shown as potential driver of COVID-19 in humans. Though most studies in animal models describe the pulmonary lesions characterized by interstitial inflammation, type II pneumocyte hyperplasia, edema, fibrin formation and infiltration of macrophages and neutrophils, detailed pathological description of vascular lesions or NETs in COVID-19 animal models are lacking so far. Here we report different types of pulmonary vascular lesions in the golden Syrian hamster model of COVID-19. Vascular lesions included endothelialitis and vasculitis at 3 and 6 days post infection (dpi), and were almost nearly resolved at 14 dpi. Importantly, virus antigen was present in pulmonary lesions, but lacking in vascular alterations. In good correlation to these data, NETs were detected in the lungs of infected animals at 3 and 6 dpi. Hence, the Syrian hamster seems to represent a useful model to further investigate the role of vascular lesions and NETs in COVID-19 pathogenesis.
Neuroimmune Consequences of eIF4E Phosphorylation on Chemotherapy-Induced Peripheral Neuropathy
Authors: Agalave Nilesh M. , Mody Prapti H. , Szabo-Pardi Thomas A. , Jeong Han S. , Burton Michael D.
Published: April 2021

Chemotherapy-induced peripheral neuropathy (CIPN) is a major dose-limiting side effect that occurs in up to 63% of patients and has no known effective treatment. A majority of studies do not effectively assess sex differences in the onset and persistence of CIPN. Here we investigated the onset of CIPN, a point of therapeutic intervention where we may limit, or even prevent the development of CIPN. We hypothesized that cap-dependent translation mechanisms are important in early CIPN development and the bi-directional crosstalk between immune cells and nociceptors plays a complementary role to CIPN establishment and sex differences observed. In this study, we used wild type and eIF4E-mutant mice of both sexes to investigate the role of cap-dependent translation and the contribution of immune cells and nociceptors in the periphery and glia in the spinal cord during paclitaxel-induced peripheral neuropathy. We found that systemically administered paclitaxel induces pain-like behaviors in both sexes, increases helper T-lymphocytes, downregulates cytotoxic T-lymphocytes, and increases mitochondrial dysfunction in dorsal root ganglia neurons; all of which is eIF4E-dependent in both sexes. We identified a robust paclitaxel-induced, eIF4E-dependent increase in spinal astrocyte immunoreactivity in males, but not females. Taken together, our data reveals that cap-dependent translation may be a key pathway that presents relevant therapeutic targets during the early phase of CIPN. By targeting the eIF4E complex, we may reduce or reverse the negative effects associated with chemotherapeutic treatments.
Improvement of Myelopoiesis in Cyclophosphamide-Immunosuppressed Mice by Oral Administration of Viable or Non-Viable Lactobacillus Strains
Authors: Gramajo Lopez Andrés , Gutiérrez Florencia , Saavedra Lucila , Hebert Elvira Maria , Alvarez Susana , Salva Susana
Published: April 2021

Myelosuppression is the major dose-limiting toxicity of cancer chemotherapy. There have been many attempts to find new strategies that reduce myelosuppression. The dietary supplementation with lactic acid bacteria (LAB) improved respiratory innate immune response and the resistance against respiratory pathogens in immunosupressed hosts. Although LAB viability is an important factor in achieving optimal protective effects, non-viable LAB are capable of stimulating immunity. In this work, we studied the ability of oral preventive administration of viable and non-viable Lactobacillus rhamnosus CRL1505 or L. plantarum CRL1506 (Lr05, Lr05NV, Lp06V or Lp06NV, respectively) to minimize myelosuppressive and immunosuppressive effects derived from chemotherapy. Cyclophosphamide (Cy) impaired steady-state myelopoiesis in lactobacilli-treated and untreated control mice. Lr05V, Lr05NV and Lp06V treatments were the most effective to induce the early recovery of bone marrow (BM) tissue architecture, leukocytes, myeloid, pool mitotic and post-mitotic, peroxidase positive, and Gr-1Low/High cells in BM. We selected the CRL1505 strain for being the one capable of maintaining its myelopoiesis-enhancing properties in its non-viable form. Although the CRL1505 treatments do not modify the Cy ability to induce apoptosis, both increased the incorporation of BrdU in BM cells. Consequently, Lr05NV and Lr05V treatments were able to promote early recovery of LSK cells (Lin-Sca-1+c-Kit+ cells), multipotent progenitors (Lin-Sca-1+c-Kit+CD34+ cells), and myeloid cells (Gr-1+Ly6G+Ly6C- cells) with respect to the untreated Cy control. In addition, these treatments were able to increase the frequency of IL17A-producing innate lymphoid cells in the intestinal lamina propria (IL-17A+RORγt+CD4-NKp46+ cells) after Cy injection. These results were correlated with an increase in the IL-17A serum levels, a GM-CSF high expression and a CXCL12 lower expression in BM. Therefore, both Lr05V and Lr05NV treatments are able to activate beneficially the IL-17A/GM-CSF axis and accelerate the recovery of Cy-induced immunosuppression by increasing BM myeloid precursors. We demonstrated for the first time the beneficial effect of CRL1505 strain on myelopoiesis affected by a chemotherapeutic drug. Furthermore, Lr05NV could be a good and safe resource for reducing chemotherapy-induced leukopenia. The results are a starting point for future research and open up broad prospects for future applications of the immunobiotics.
Aberrant Gut Microbiome Contributes to Intestinal Oxidative Stress, Barrier Dysfunction, Inflammation and Systemic Autoimmune Responses in MRL/lpr Mice
Authors: Wang Hui , Wang Gangduo , Banerjee Nivedita , Liang Yuejin , Du Xiaotang , Boor Paul J. , Hoffman Kristi L. , Khan M. Firoze
Published: April 2021

Microbiome composition and function have been implicated as contributing factors in the pathogenesis of autoimmune diseases (ADs), including systemic lupus erythematosus (SLE), rheumatoid arthritis and autoimmune hepatitis (AIH). Furthermore, dysbiosis of gut microbiome is associated with impaired barrier function and mucosal immune dysregulation. However, mechanisms by which gut microbiome contributes to the ADs and whether antioxidant treatment can restore gut homeostasis and ameliorate the disease outcome are not known. This study was, therefore, focused on examining the involvement of gut microbiome and host responses in the pathogenesis of SLE using unique female mouse models (C57BL/6, MRL+/+ and MRL/lpr) of 6 and 18 weeks with varying degrees of disease progression. Fecal microbiome diversity and composition, gut oxidative stress (OS), barrier function and inflammation, as well as systemic autoimmunity were determined. Interestingly, each mouse strain had distinct bacterial community as revealed by β-diversity. A lower Firmicutes/Bacteroidetes ratio in 6-week-old MRL/lpr mice was observed, evidenced by decrease in Peptostreptococcaceae under Firmicutes phylum along with enrichment of Rikenellaceae under Bacteroidetes phylum. Additionally, we observed increases in colonic OS [4-hydroxynonenal (HNE)-adducts and HNE-specific immune complexes], permeability changes (lower tight junction protein ZO-2; increased fecal albumin and IgA levels) and inflammatory responses (increased phos-NF-κB, IL-6 and IgG levels) in 18-week-old MRL/lpr mice. These changes were associated with markedly elevated AD markers (antinuclear and anti-smooth muscle antibodies) along with hepatic portal inflammation and severe glomerulonephritis. Notably, antioxidant N-acetylcysteine treatment influenced the microbial composition (decreased Rikenellaceae; increased Akkeransiaceae, Erysipelotrichaceae and Muribaculaceae) and attenuated the systemic autoimmunity in MRL/lpr mice. Our data thus show that gut microbiome dysbiosis is associated with increased colonic OS, barrier dysfunction, inflammatory responses and systemic autoimmunity markers. These findings apart from delineating a role for gut microbiome dysbiosis, also support the contribution of gut OS, permeability changes and inflammatory responses in the pathogenesis of ADs.
Tributyrin Plays an Important Role in Regulating the Growth and Health Status of Juvenile Blunt Snout Bream (Megalobrama amblycephala), as Evidenced by Pathological Examination
Authors: Liang Hualiang , Ji Ke , Ge Xianping , Xi Bingwen , Ren Mingchun , Zhang Lu , Chen Xiaoru
Published: April 2021

The present study aimed to assess the role of tributyrin (TB) in regulating the growth and health status of juvenile blunt snout bream (Megalobrama amblycephala) through an 8-week feeding experiment. Six groups were fed experimental diets with added TB percentages of 0% (control group), 0.03%, 0.06%, 0.09%, 0.12% and 0.15%. The present results showed that TB supplementation in feed had some positive impacts on FW, WG, FCR and SGR, and the best results were found in the 0.06% TB group (P<0.05). However, TB supplementation in feed had no significant effects on SR, CF, VSI or whole-body composition (P>0.05). TB supplementation in feed increased antioxidant capacity and immunological capacity and attenuated the inflammatory response by increasing the activity of T-SOD, GPx, CAT and the levels of anti-inflammatory cytokines (IL-10 and TGF-β) and decreasing the levels of MDA and anti-inflammatory cytokines (TNF-α) (P<0.05). Furthermore, TB supplementation improved immunity by increasing the levels of immunoglobulins (IgM and IgG), C3 and IFN-γ (P<0.05). Surprisingly, 0.06%-0.12% TB supplementation significantly increased the content of IL-1β (P<0.05). However, TB supplementation in feed had no significant effects on the plasma content of GSH, HSP70, IL-8 and the activity of T-AOC (P>0.05). The possible mechanism was that TB activated PI3K/Akt/Nrf2 and inhibits the NF-κB signaling pathway, further regulating the mRNA levels of key genes with antioxidant capacity and the inflammatory response; for example, it increased the mRNA levels of Nrf2, Cu/Zn-SOD, HO-1, CAT, Akt, PI3K, GPx, IL-10, and TGF-β and decreased the mRNA levels of NF-κB and TNF-α (P<0.05). In addition, 0.06%-0.15% TB supplementation significantly increased the mRNA levels of IL-1β (P<0.05). TB supplementation in feed had no significant effects on the mRNA levels of HSP70, Mn-SOD and IL-8 (P>0.05). Evidence was presented that TB supplementation decreased the mortality rate caused by Aeromonas hydrophila challenge. In pathological examination, TB supplementation prevented hepatic and intestinal damage. Generally, TB supplementation improved the growth performance of juvenile blunt snout bream. Furthermore, TB supplementation activated PI3K/Akt/Nrf2 and inhibited the NF-κB signaling pathway, regulating health status and preventing hepatic and intestinal damage.
Using Routine Laboratory Markers and Immunological Indicators for Predicting Pneumocystis jiroveci Pneumonia in Immunocompromised Patients
Authors: Tang Guoxing , Tong Shutao , Yuan Xu , Lin Qun , Luo Ying , Song Huijuan , Liu Wei , Wu Shiji , Mao Liyan , Liu Weiyong , Zhu Yaowu , Sun Ziyong , Wang Feng
Published: April 2021

Background

Pneumocystis jiroveci pneumonia (PJP) is the most common opportunistic infection in immunocompromised patients. The accurate prediction of PJP development in patients undergoing immunosuppressive therapy remains challenge.

Methods

Patients undergoing immunosuppressive treatment and with confirmed pneumocystis jiroveci infection were enrolled. Another group of matched patients with immunosuppressant treatment but without signs of infectious diseases were enrolled to control group.

Results

A total of 80 (40 PJP, 40 non-PJP) participants were enrolled from Tongji Hospital. None of the patients were HIV positive. The routine laboratory indicators, such as LYM, MON, RBC, TP, and ALB, were significantly lower in PJP patients than in non-PJP patients. Conversely, LDH in PJP patients was significantly higher than in non-PJP controls. For immunological indicators, the numbers of T, B, and NK cells were all remarkably lower in PJP patients than in non-PJP controls, whereas the functional markers such as HLA-DR, CD45RO and CD28 expressed on CD4+ or CD8+ T cells had no statistical difference between these two groups. Cluster analysis showing that decrease of host immunity markers including CD3+, CD4+ and CD8+ T cells, and increase of tissue damage marker LDH were the most typical characteristics of PJP patients. A further established model based on combination of CD8+ T cells and LDH showed prominent value in distinguishing PJP from non-PJP, with AUC of 0.941 (95% CI, 0.892-0.990).

Conclusions

A model based on combination of routine laboratory and immunological indicators shows prominent value for predicting the development of PJP in HIV-negative patients undergoing immunosuppressive therapy.