bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cryopreserved Human Adipose-derived Stromal Vascular Fraction Maintains Fracture Healing Potential via Angiogenesis and Osteogenesis in an Immunodeficient Rat Model
Authors: Kamenaga Tomoyuki , Kuroda Yuichi , Nagai Kanto , Tsubosaka Masanori , Takashima Yoshinori , Kikuchi Kenichi , Fujita Masahiro , Ikuta Kemmei , Anjiki Kensuke , Maeda Toshihisa , Nakano Naoki , Takayama Koji , Hashimoto Shingo , Hayashi Shinya , Matsushita Takehiko , Niikura Takahiro , Kuroda Ryosuke , Matsumoto Tomoyuki 
Published: August 2020

Abstract
Background: Novel therapeutic strategies for the healing of nonunion, which has serious effects on the quality of life of patients, are needed. We evaluated the therapeutic effect of local transplantation of human stromal vascular fraction (SVF) cells on fracture healing in a rat non-healing fracture model and compared the effects between freshly isolated (F) and cryopreserved (C)-SVFs. Methods: Nonhealing fracture model was induced in the femur of female immunodeficient rats (F344/N Jcl rnu/rnu) with cauterizing periosteum. Immediately after the creation of non-healing fracture, rats received local transplantation of F and C-SVFs suspended in phosphate-buffered saline (PBS) or the same volume of PBS without cells using the same scaffold as a control group. During 8 weeks post-surgery, radiologic, histological, immunohistochemical, and biomechanical analyses were performed to evaluate fracture healing. The comparison of radiological results was performed with a chi-square test, and the multiple comparisons of immunohistochemical, histological and biomechanical results among groups were made using a one-way analysis of variance. A probability value of 0.05 was considered to denote statistical significance.Results: At week 8, in 60% of animals receiving F-SVF cells and in 50% of animals receiving C-SVF cells, the fracture radiologically healed with bone union whereas nonunion was observed in the control group. The healing potential was also confirmed by histological and biomechanical assessments. One of the mechanisms of healing involving intrinsic angiogenesis/osteogenesis was enhanced in F- and C-SVF groups compared with that in the control group. Human cells-derived vasculogenesis/osteogenesis, which was also confirmed in an in vitro differentiation assay, was also enhanced in the F- and C-SVF groups compared with that in the control groups, and could be another mechanism for healing. Conclusions: SVF cells can enhance bone healing and cryopreserved cells have almost equal potential as fresh cells. SVF cells can be used for improving nonunion bone fracture healing as an alternative to other mesenchymal stem cells and the effect of SVF cells can be maintained under cryopreservation.
The foodborne diseases antimicrobial resistance development in food animals: A case of Salmonella isolates from diarrheic sheep in and around Gondar city, Ethiopia: A cross-sectional study
Authors: Gebeyehu Daniel Teshome 
Published: August 2020

Abstract
Background: In recent years, an increase in the occurrence of antimicrobial-resistant pathogens especially foodborne zoonotic bacteria has been observed in several countries. As a result, fatality rates are increasing due to those resistant bacteria in both human and animal populations, particularly in developing countries like Ethiopia where the risk of infection is high due to poor biosecurity measures, close animal-human interactions, and extensive use of antimicrobials for animal productions. One of those zoonotic diseases, which commonly contaminates food and having zoonotic characteristics is Salmonella.Methodology: A cross-sectional study on samples collected from diarrheic sheep in and around Gondar city, to assess the risk of antibiotic resistance development of Salmonella in food animal production was conducted. A total of 80 positively isolated Salmonellae were taken from 165 diarrheic sheep and antibiotics resistance test was conducted by Kirby-Bauer Disc diffusion method. The disc diffusion test was performed using ten commonly used antibiotic discs in the study area.Result: Salmonellae were 100% resistant for amoxicillin and ampicillin. In addition to this, the bacteria were sensitive for kanamycin, gentamicin, and ciprofloxacin with some level of intermediate resistance. Salmonellae were 75% sensitive with 25% resistance for chloramphenicol while trimethoprim, tetracycline, doxycycline and nalidixic acid showed 37.5%, 37.5%, 25% and 12.5% of resistance respectively. Generally, this study revealed that Salmonellae were developed a wide range of resistance to different antibiotics in the study area.Conclusion: In general, majority of previously curative antibiotics are developed resistance against Salmonella. Therefore, the rational use of antibiotics, antimicrobial use governance in animal production, further studies, and integrative approaches among animal-human health professionals are recommended for the reduction and mitigation of health risks arising from antibiotic-resistant zoonotic pathogens like Salmonellae.
A signature of seven immune-related genes predicts overall survival in male gastric cancer patients
Authors: Xu Xin , Wu Youliang , Wang Mingliang , Lu Yida , Wang Xiaodong , Wang Huizhen , Chen Bo , Li Yongxiang 
Published: August 2020

Abstract
Background: Gastric cancer (GC) is one of the most fatal malignant tumors with a high mortality rate. Male sex has been proven as an independent risk factor for GC. This study aimed to identify immune-related genes (IRGs) for the prognosis of male GC.Method: RNA sequencing and clinical data were obtained from The Cancer Genome Atlas (TCGA) database. Differentially expressed IRGs between male GC and normal tissues were identified by integrated bioinformatics analysis. Univariate and multivariate Cox regression analysis were applied to screen survival-associated IRGs. Then, GC patients were separated into high- and low-risk groups based on the median risk score. Furthermore, a nomogram was constructed based on the TCGA dataset. The prognostic value of the risk signature model was evaluated by Kaplan-Meier curve, receiver operating characteristic (ROC), Harrell’s concordance index and calibration curves. In addition, the gene expression dataset from Gene Expression Omnibus (GEO) was also downloaded for external validation. The relative proportions of 22 types of infiltrating immune cells in each male GC sample were evaluated using CIBERSORT.Results: A total of 276 differentially expressed IRGs were screened, including 189 up-regulated and 87 down-regulated genes. Subsequently, a seven-IRGs signature (LCN12, CCL21, RNASE2, CGB5, NRG4, AGTR1 and NPR3) was identified and showed a significant association with the overall survival (OS) of male GC patients. Survival analysis indicated that patients with high-risk group exhibited a poor clinical outcome. The result of multivariate analysis revealed that the risk score was an independent prognostic factor. The established nomogram could be used to evaluate the prognosis of individual male GC patients. Further analysis showed that the prognostic model had an excellent predictive performance in both TCGA and validated cohorts. Besides, the result of tumor-infiltrating immune cell analysis indicated that the seven-IRGs signature could reflect the status of the tumor immune microenvironment.Conclusions: Our study developed a novel seven-IRGs risk signature for individualized survival prediction of male GC patients.
Bone marrow mesenchymal stem cell-derived exosomes promote rotator cuff tendon-bone healing by promoting angiogenesis and regulating M1 macrophages in rats
Authors: Huang Yao , He Bing , Wang Lei , Yuan Bin , Shu Hao , Zhang Fucheng , Sun Luning 
Published: August 2020

Abstract
Background: Rotator cuff tears (RCTs) often require reconstructive surgery. Tendon-bone healing is critical for the outcome of rotator cuff reconstruction, but the process of tendon-bone healing is complex and difficult. Mesenchymal stem cells (MSCs) are considered to be an effective method to promote tendon-bone healing. MSCs have strong paracrine, anti-inflammatory, immunoregulatory, and angiogenic potential. Recent studies have shown that MSCs achieve many regulatory functions through exosomes. The purpose of this study was to explore the role of bone MSC-derived exosomes (BMSC-Exos) in tendon-bone healing. Methods: Our study found that BMSC-Exos promote the proliferation, migration, and angiogenic tube formation of Human Umbilical Vein Endothelial Cells (HUVECs). The mechanism by which BMSC-Exos achieve this may be through the regulation of the angiogenic signaling pathway. In addition, BMSC-Exos can inhibit the polarization of M1 macrophages and inhibit the secretion of proinflammatory factors by M1 macrophages. After rotator cuff reconstruction in rats, BMSC-Exos were injected into the tail vein to analyze their effect on the rotator cuff tendon-bone interface healing. Results: It was confirmed that BMSC-Exos increased the breaking load and stiffness of the rotator cuff after reconstruction in rats, induced angiogenesis around the rotator cuff endpoint, and promoted growth of the tendon-bone interface. Conclusion: BMSC-Exos promote tendon-bone healing after rotator cuff reconstruction in rats by promoting angiogenesis and inhibiting inflammation.
Effect of the Pumping-injection flow rate on Heat Transfer Characteristic of Borehole Heat Exchangers for Coupling Ground-Source Heat Pump System
Authors: Ma Jiuchen , Jiang Qian , Zhang Qiuli , Xie Yacheng , Wang Yahui , Yi Feiyu 
Published: August 2020

Abstract
A coupling ground source heat pump system (CGSHP) is established in areas where groundwater is shallow but the seepage velocity is weak, which sets up pumping and injection wells on both sides of borehole heat exchangers (BHEs). A convection-dispersion analytical model of excess temperature in aquifer that considers groundwater forced seepage and axial effects and thermal dispersion effects is proposed. A controllable forced seepage sandbox is built by equation analysis method and similarity criteria. Through indoor test and the proposed analytical model, the correctness and accuracy of the numerical simulation software FEFLOW7.1 is verified. The influence of different pumping-injection flow rate on the heat transfer characteristic of BHEs is studied by numerical simulation. The results show that the average heat efficiency coefficient of BHEs increases and the heat influence range of downstream BHEs expands with the increasing of pumping-injection flow rate. The relation curve between Pe and the increment of heat transfer rate per unit depth of BHEs (Δ‾ q ) is distributed as Gaussian function. The pumping-injection flow rate that makes Darcy velocity reaches 0.6×10 -6 ~1.4×10 -6 m∙s -1 in the aquifer is the best reference range for CGSHP system,so 400~600 m 3 ∙d -1 is taken as the best pumping-injection flow rate in this paper.
Construction of a new T7 promoter compatible Escherichia coli Nissle 1917 strain for recombinant production of heme-dependent proteins
Authors: Kiege Kerstin , Frankenberg-Dinkel Nicole 
Published: August 2020

Abstract
Background: Heme proteins and heme-derived molecules play an important role in several cellular processes. Therefore, their production and functional analysis is of great research interest. For the analysis of these molecules, high production yields are required. We recently reported the use of the probiotic E. coli strain Nissle 1917 (EcN) to sufficiently produce heme proteins. This strain is capable of taking up heme from the growth medium due to the outer membrane heme receptor ChuA which is absent in regular E. coli expression strains. Unfortunately, the strain lacks the gene for T7 RNA polymerase which is necessary for the expression of genes under the control of the T7-promotor, widely used in expression vectors like the pET or Duet series.
Results: A new T7-promoter compatible EcN strain was constructed. Therefore, the gene for T7-RNA polymerase under the control of a lac UV5 promoter was integrated into the malEFG operon of EcN. Test expressions of genes via T7 promoter-based vectors in the new EcN(T7) strain were successful. Expression in EcN(T7) efficiently resulted in the production of recombinant heme proteins in which the heme cofactor was incorporated during protein production. In addition, the new EcN(T7) strain can be used to co-express genes for the production of heme-derived molecules like biliverdin or other open-chain tetrapyrroles. We demonstrate successful recombinant production of the phytochromes BphP from Pseudomonas aeruginosa and Cph1 from Synechocystis sp. PCC6803 loaded with their cofactor biliverdin and phycocyanobilin, respectively.
Conclusion: We present a new E. coli strain for sufficient production of heme proteins and heme-derived molecules using T7-promoter based expression vectors. The new EcN(T7) strain enables the use of a broader spectrum of expression vectors as well as the co-expression of genes using the Duet expression vectors. Furthermore, the capability of feeding EcN and EcN(T7) with heme overcomes the rate limiting step in the recombinant heme protein production, i.e. heme biosynthesis of E. coli . Therefore, a higher heme saturation of heme proteins and also higher yields of heme-derived molecules is obtained using the constructed strain.
The Temperature of Growth and Sporulation Modulates the Efficiency of Spore-Display in Bacillus Subtilis
Authors: Petrillo Claudia , Castaldi Stefany , Lanzilli Mariamichela , Saggese Anella , Donadio Giuliana , Baccigalupi Loredana , Ricca Ezio , Isticato Rachele 
Published: August 2020

Abstract
Background: Bacterial spores displaying heterologous antigens or enzymes have long been proposed as mucosal vaccines, functionalized probiotics or biocatalysts. Two main strategies have been developed to display heterologous molecules on the surface of Bacillus subtilis spores: i) a recombinant approach, based on the construction of a gene fusion between a gene coding for a coat protein (carrier) and DNA coding for the protein to be displayed, and ii) a non-recombinant approach, based on the spontaneous and stable adsorption of heterologous molecules on the spore surface. Both systems have advantages and drawbacks and the selection of one or the other depends on the protein to be displayed and on the final use of the activated spore. It has been recently shown that B. subtillis builds structurally and functionally different spores when grown at different temperatures, based on that B. subtilis spores prepared at 25, 37 or 42°C were compared for their efficiency in displaying various model proteins by either the recombinant or the non-recombinant approach.Results: Immune- and fluorescence-based assays were used to analyze the display of several model proteins on spores prepared at 25, 37 or 42°C. Recombinant spores displayed different amounts of the same fusion protein in response to the temperature of spore production. In spores simultaneously displaying two fusion proteins, each of them was differentially displayed at the various temperatures. The display by the non-recombinant approach was only modestly affected by the temperature of spore production, with spores prepared at 37 or 42°C slightly more efficient than 25°C spores in adsorbing at least some of the model proteins tested. Conclusion: Our results indicate that the temperature of spore production allows to control the display of heterologous proteins on spores and, therefore, that the spore-display strategy can be optimized for the specific final use of the activated spores by selecting the display approach, the carrier protein and the temperature of spore production.
Characterization of florfenicol resistance genes in the coagulase-negative Staphylococcus (CoNS) isolates and genomic features of a multidrug-resistant Staphylococcus lentus strain H29
Authors: Wu Chongyang , Zhang Xueya , Liang Jialei , Li Qiaoling , Lin Hailong , Lin Chaoqing , Liu Hongmao , Zhou Danying , Lu Wei , Sun Zhewei , Lin Xi , Zhang Hailin , Li Keiwei , Xu Teng , Bao Qiyu , Lu Junwan 
Published: August 2020

Abstract
BackgroundWith the wide use of florfenicol to prevent and treat the bacterial infection of domestic animals, the emergence of the florfenicol resistance bacteria is increasingly serious. It is very important to elucidate the molecular mechanism of the bacteria’s resistance to florfenicol. MethodsThe minimum inhibitory concentration (MIC) levels was determined by the agar dilution method, and polymerase chain reaction (PCR) was conducted to analyze the distribution of florfenicol resistance genes in 39 CoNS strains isolated from poultry and livestock animals and seafood. The whole genome sequence of one multidrug-resistant strain, Staphylococcus lentus H29, was characterized, and comparative genomics analysis of the resistance gene-related sequences was also performed. ResultsAs a result, the isolates from the animals showed a higher resistance rate (23/28, 82.1%) and much higher MIC levels of florfenicol than those from seafood. Twenty-seven animal isolates carried 37 florfenicol resistance genes (including 26 fexA , 6 cfr and 5 fexB genes), of which 1 carried a cfr gene, 16 carried a fexA gene, 5 carried both fexA and fexB genes and 5 carried both fexA and cfr genes. On the other hand, all 11 isolates from seafood were sensitive to florfenicol, and only 3 carried a fexA gene each. The whole genome sequence of S. lentus H29 was composed of a chromosome and two plasmids ( pH29-46 , pH29- 26) and harbored 11 resistance genes, including 6 genes [ cfr, fexA, ant(6)-Ia , aac A -aph D , mecA and mph(C) ] encoded on the chromosome, four genes [ cfr, fexA, aac A -aph D and tcaA ] on pH29-46 and one gene ( fosD ) on pH29-26. It was interested to find that the S. lentus H29 genome carried two identical copies of the gene arrays of radC - tnpABC - hp - fexA (5,671 bp) and IS 256 - cfr (2,690 bp), of which one copy of the two gene arrays was encoded on plasmid pH29-46, while the other was encoded on the chromosome. ConclusionsThe current study revealed the wide distribution of florfenicol resistance genes ( cfr, fexA and fexB ) in animal bacteria, and to the best of our knowledge, this is the first report of one CoNS strain carrying two identical copies of florfenicol resistance-related gene arrays.
Effect of early versus delayed or no intubation on clinical outcomes of patients with COVID-19&nbsp;
Authors: Siempos Ilias , Xourgia Elena , Ntaidou Theodora K. , Zervakis Dimitris , Magira Eleni E. , Kotanidou Anastasia , Routsi Christina , Zakynthinos Spyros G. 
Published: August 2020

Abstract
Background: Optimal timing of initiation of invasive mechanical ventilation in patients with acute hypoxemic respiratory failure due to COVID-19 is unknown. Thanks to early flattening of the epidemiological curve, ventilator demand in Greece was kept lower than supply throughout the pandemic, allowing for unbiased comparison of the outcomes of patients undergoing early intubation versus delayed or no intubation.Methods: We conducted an observational study including all adult patients with laboratory-confirmed COVID-19 consecutively admitted in Evangelismos Hospital, Athens, Greece between March 11, 2020 and April 15, 2020. Patients subsequently admitted in the intensive care unit (ICU) were categorized into the “early intubation” versus the “delayed or no intubation” group.Results: During the study period, a total of 101 patients (37% female, median age 65 years) were admitted in the hospital. Fifty-nine patients (58% of the entire cohort) were exclusively hospitalized in general wards with a mortality of 3% and median length of stay of 7 days. Forty-two patients (19% female, median age 65 years, 62% with at least one comorbidity, 14% never intubated) were admitted in the ICU; all with acute hypoxemic respiratory failure. Early intubation was not associated with higher ICU-mortality (21% versus 33%), fewer ventilator-free days (3 versus 2 days) or fewer ICU-free days than delayed or no intubation.Conclusions: A strategy of early intubation was not associated with worse clinical outcomes compared to delayed or no intubation. Given that early intubation may presumably reduce virus aerosolization, these results may justify further research with a randomized controlled trial.
Postpartum dyslipidemia and the potential predictors in women with history of gestational diabetes mellitus
Authors: Pei Ling , Xiao Huangmeng , Lai Fenghua , Li Zeting , Li Zhuyu , Yue Shufan , Chen Haitian , Li Yanbin , Cao Xiaopei 
Published: August 2020

Abstract
Aims/Introduction: To analyze the incidence of early postpartum dyslipidemia and the potential predictors among women with history of gestational diabetes mellitus (GDM).Materials and Methods: This was a retrospective study. 589 women diagnosed with GDM were recruited and followed up at 6-12 weeks after delivery. The general demographic and metabolic data during pregnancy were collected. Participants were divided into the normal lipid group and dyslipidemia group according to the postpartum lipid level. Demographic and metabolic parameters were compared. Multivariate logistic regression was performed to analyze the potential predictors for the early postpartum dyslipidemia. Receiver operating characteristic (ROC) curve was conducted to determine the cut-off values.Results: 38.5% of the 589 women developed dyslipidemia in early postpartum and 60% of them had normal glucose metabolism. Delivery age, systolic blood pressure (SBP), glycated hemoglobin (HbA1c) and low-density lipoprotein cholesterol (LDL-C) were independent predictors of early postpartum dyslipidemia in women with history of GDM. The cut-offs of maternal age, SBP, HbA1c and LDL-C were 35 years, 5.1% and 3.56 mmol/L, respectively. LDL-C achieved a balanced mix of high sensitivity（63.9%）and specificity(69.2%）with highest area under the receiver operating characteristic curve（AUC) (0.696). When LDL-C was combined with age, SBP and HbA1c, the AUC reached to 0.733.Conclusions: Women with history of GDM should be screened lipid metabolism after delivery, particularly those with maternal age >35 years, SBP>123mmHg before labor, HbA1c>5.1%, or LDL-C>3.56mmol/L in the second trimester of pregnancy.