bluuebunny/crossref_metadata_2025 huggingface dataset:
Uncovering the Genomic Potential of the Amazon River Microbiome to Degrade Rainforest Organic Matter
Authors: Júnior Célio Dias Santos , Sarmento Hugo , de Miranda Fernando Pellon , Henrique-Silva Flávio , Logares Ramiro
Published: August 2020

Background: The Amazon River is one of the largest in the world and receives huge amounts of terrestrial organic matter (TeOM) from the surrounding rainforest. Despite this TeOM is typically recalcitrant (i.e. resistant to degradation), only a small fraction of it reaches the ocean, pointing to a substantial TeOM degradation by the river microbiome. Yet, microbial genes involved in TeOM degradation in the Amazon River were barely known. Here, we examined the Amazon River microbiome by analyzing 106 metagenomes from 30 sampling points distributed along the river.Results: We constructed the Amazon River basin Microbial non-redundant Gene Catalogue (AMnrGC) that includes ~3.7 million non-redundant genes, affiliating mostly to bacteria. We found that the Amazon River microbiome contains a substantial gene-novelty compared to other relevant known environments (rivers and rainforest soil). Genes encoding for proteins potentially involved in lignin degradation pathways were correlated to tripartite tricarboxylates transporters and hemicellulose degradation machinery, pointing to a possible priming effect. Based on this, we propose a model on how the degradation of recalcitrant TeOM could be modulated by labile compounds in the Amazon River waters. Our results also suggest changes of the microbial community and its genomic potential along the river course.Conclusions: Our work contributes to expand significantly our comprehension of the world’s largest river microbiome and its potential metabolism related to TeOM degradation. Furthermore, the produced gene catalogue (AMnrGC) represents an important resource for future research in tropical rivers.
The Effects of Resistance Training With or Without Peanut Protein Supplementation on Skeletal Muscle and Strength Adaptations in Older Individuals
Authors: Lamb Donald A. , Moore Johnathon H. , Smith Morgan A. , Vann Christopher G. , Osburn Shelby C. , Ruple Bradley A. , Fox Carlton D. , Smith Kristen S. , Altonji Olivia M. , Power Zade M. , Cerovsky Annsley E. , Ross C. Owen , Cao Andy T. , Goodlett Michael D. , Huggins Kevin W. , Fruge Andrew D. , Young Kaelin C. , Roberts Michael D.
Published: August 2020

Several studies suggest resistance training (RT) with protein supplementation has positive effects on strength and muscle mass in older individuals. However, to date, no study has examined the effects of RT with a high-protein, defatted peanut powder (PP) supplement on these markers. Herein, 39 older, untrained individuals (n=17 female, n=22 male; age=58.6±8.0 years; body mass index =28.7±5.8) completed a 6-week (n=22) or 10-week (n=17) RT program, where full-body training was implemented twice weekly (ClinicalTrials.gov trial registration NCT04015479; registered July 11, 2019). Participants in each program were randomly assigned to consume either a PP supplement once per day (35 g protein, 315 kcal; n=20) or no supplement (CTL; n=19). Right leg vastus lateralis (VL) muscle biopsies were obtained prior to and 24 hours following the first training bout in all participants to assess the change in myofibrillar protein synthetic rates (MyoPS) as measured via the deuterium-oxide (D2O) tracer method. Pre- and Post-intervention testing in all participants was conducted using dual energy x-ray absorptiometry (DXA), VL ultrasound imaging, a peripheral quantitative computed tomography (pQCT) scan at the mid-thigh, and right leg isokinetic dynamometer assessments. Integrated MyoPS rates over a 24-hour period were not significantly different (p<0.05) between supplement groups following the first training bout. Regarding chronic changes, there were no significant supplement-by-time interactions in DXA-derived fat mass, lean soft tissue mass or percent body fat between supplementation groups. There was, however, a significant increase in VL thickness in PP versus CTL participants when the 6- and 10-week cohorts were pooled (interaction p=0.041). There was also a significant increase in knee flexion torque in the 10-week PP group versus the CTL group (interaction p=0.032). In conclusion, a high-protein, defatted peanut powder supplement in combination with RT positively affects select markers of muscle hypertrophy and strength in an untrained, older adult population.
Laboratory and microcosm experiments reveal contrasted adaptive responses to ammonia and water mineralisation in aquatic stages of the sibling species Anopheles gambiae s.s. and Anopheles coluzzii
Authors: Akpodiete Nwamaka Oluchukwu , Tripet Frederic
Published: August 2020

Background The sibling species of the malaria mosquito, Anopheles gambiae s.s and Anopheles coluzzii co-exist in many parts of West Africa and are thought to have recently diverged through a process of ecological speciation with gene flow. Divergent larval ecological adaptations, resulting in Genotype × Environment interactions have been proposed as important drivers of speciation in these species. In West Africa, An. coluzzii tends to be associated with permanent man-made larval habitats such as irrigated rice fields which are typically more eutrophic and mineral and ammonia-rich than the temporary rain pools exploited by An. gambiae s.s. Methods To highlight G×E interactions at the larval stage and their possible role in ecological speciation of these species, we first investigated the effect of exposure to ammonium hydroxide and water mineralisation on larval developmental success. Mosquito larvae were exposed to two water sources and increasing ammonia concentrations in small containers until adult emergence. In a second experiment, larval developmental success was compared across 2 contrasted microcosms to highlight G × E interactions under conditions like those found in the natural environment. Results The first experiment revealed significant G × E interactions in developmental success and phenotypic quality for both species in response to increasing ammonia concentrations and water mineralisation. The An. coluzzii strain outperformed the An. gambiae s.s. strain under limited conditions that were closer to more eutrophic habitats. The second experiment revealed divergent crisscrossing reaction norms in the developmental success of the sibling species in the two contrasted larval environments. As expected, An. coluzzii had higher emergence rates in the rice paddy environment with emerging adults of superior phenotypic quality compared to An. gambiae s.s, and vice versa in the rain puddle environment.Conclusions Evidence for such G x E interactions lends support to the hypothesis that divergent larval adaptations to the environmental conditions found in man-made habitats such as rice-fields in An. coluzzii may have been an important driver of its ecological speciation.
Regular Consumption of Soft Drinks is Associated with Type 2 Diabetes Incidence in Mexican Adults: Findings from a Prospective Cohort Study.
Authors: Torres-Ibarra Leticia , Rivera-Paredez Berenice , Hernández-López Rubí , Canto-Osorio Francisco , Sánchez-Romero Luz María , López-Olmedo Nancy , González Romina , Ramírez Paula , Salmerón Jorge , Barrientos-Gutiérrez Tonatiuh
Published: August 2020

Background: Although high consumption of soft drinks has been associated with excess of type 2 diabetes risk, the strength of this association in the Mexican population, where a type 2 diabetes genetic susceptibility has well established, has been scarcely studied. This study aimed to estimate the risk of type 2 diabetes due to soft drinks consumption in a cohort of Mexicans. Methods: We used data on 1,445 participants from the Health Workers Cohort Study, a prospective cohort conducted in Cuernavaca, Mexico. Soft drinks consumption was assessed with a semi-quantitative 116-item food frequency questionnaire. Incident type 2 diabetes was defined as self-report of physician-diagnosed type 2 diabetes, or fasting glucose >126 mg/dl, or use of hypoglycemic medication at any examination. Hazard ratios (HRs) along with 95% confidence intervals (CIs) were estimated using Cox proportional hazard models.Results: With a total of 9,526.2 person-years of follow-up, 109 incident cases of type 2 diabetes were observed. Type 2 diabetes incidence rate was 7.6, 11.0, and 17.1 per 1,000 person-years across levels of soft drinks consumption of <1, 1-4, and ³5 servings/week, respectively (p<0.001 for trend). The intake of ≥5 soft drinks/week was significantly associated with an increased risk of type 2 diabetes (HR 2.0, 95%CI: 1.1–3.8) compared with consumption of <1/week. This association was not modified by family history of diabetes. The HR was attenuated by further adjustment for body mass index (HR 1.6, 95%CI: 0.8-2.9) and abdominal obesity (HR 1.7, 95%CI: 0.9-3.2). Conclusions: The consumption of soft drinks was associated with a higher risk of type 2 diabetes in a cohort of Mexican adults. Our results further support recommendations to limit intake of soft drinks to address the growing diabetes epidemic in Mexico.
Evaluation of Differential Peptide Loading on Tandem Mass Tag-Based Proteomic and Phosphoproteomic Data Quality
Authors: Sanford James A , Wang Yang , Hansen Joshua R , Gritsenko Marina A , Weitz Karl K , Tognon Cristina E , Petyuk Vladislav A , Qian Wei-Jun , Liu Tao , Druker Brian J , Rodland Karin D , Piehowski Paul D.
Published: August 2020

Background: Global and phosphoproteome profiling has demonstrated great utility for the analysis of clinical specimens. One major barrier to the broad clinical application of proteomic profiling is the large amount of biological material required, particularly for phosphoproteomics—currently on the order of 25 mg wet tissue weight, depending on tissue type. For hematopoietic cancers such as acute myeloid leukemia (AML), the sample requirement is in excess of 10 million (1E7) peripheral blood mononuclear cells (PBMCs). Throughout the course of a prospective study, this requirement will certainly exceed what is obtainable from many of the individual patients/timepoints. For this reason, we were interested in examining the impact of differential peptide loading across multiplex channels on proteomic data quality. Methods: To achieve this, we tested a range of channel loading amounts (20, 40, 100, 200, and 400 μg of tryptic peptides, or approximately the material obtainable from 5E5, 1E6, 2.5E6, 5E6, and 1E7 AML patient cells) to assess proteome coverage, quantification reproducibility and accuracy in experiments utilizing isobaric tandem mass tag (TMT) labeling. Results: As expected, we found that fewer missing values are observed in TMT channels with higher peptide loading amounts compared to those with lower loading. Moreover, channels with lower loading amounts have greater quantitative variability than channels with higher loading amounts. Statistical analysis of the differences in means among the five loading groups showed that the 20 μg loading group was significantly different from the 400 μg loading group. However, no significant differences were detected among the 40, 100, 200 and 400 μg loading groups. Conclusions: These assessment data demonstrate the practical limits of loading differential quantities of peptides across channels in TMT multiplexes, and provide a basis for designing the optimal clinical proteomics study when specimen quantities are limited.
A three-dimensional printed model in preoperative consent for ventricular septal defect repair
Authors: Deng Xicheng , He Siping , Huang Peng , Luo Jinwen , Yang Guangxian , Zhou Bing , Xiao Yunbin
Published: August 2020

Background: The 3D printing technology in congenital cardiac surgery has been widely utilized to improve patients’ understanding of their disease. However, there has been no randomized controlled study on its usefulness in surgical consent for congenital heart disease repair.Methods: A randomized controlled study was performed during consent process in which guardians of candidates for ventricular septal defect repair were given detailed explanation of the anatomy, indication for surgery and potential complication and risks using 3D print ventricular septal defect model (n = 20) versus a conventional 2D diagram (n = 20). A questionnaire was finished by each guardian of the patients. Data collected from questionnaires as well as medical records were statistically analyzed.Results: Statistically significant improvements in ratings of understanding of ventricular septal defect anatomy (p = 0.02), and of the course of procedure and potential complications (p = 0.02) were noted in the group that used the 3D model, though there was no difference in overall ratings of the consent process (p = 0.09). There was no difference in questionnaire score between subjects with different education level. The clinical outcomes, as represented by the duration of intensive care unit stay, intubation duration was comparable between the two groups.Conclusions: The results indicated that it was an effective tool which may be used to consent for congenital heart surgery. Different education levels do not affect guardians’ understanding in consent. The impact of 3D printing used in this scenario on long term outcomes remains to be defined.
Factorial Design Study of Total Petroleum Contaminated Soil Treatment Using Land Farming Technique
Authors: Okonofua Ehizonomhen Solomon , LASISI KAYODE HASSAN , Atikpo E
Published: August 2020

Land farming technique was used to treat hydrocarbon contaminated soil collected from a crude oil spill sites in Edo State, Nigeria. Calibrated standard auger was used to collect soil samples from the site at depth below 30 cm. The samples were characterized and classified. Cow dung and NPK fertilizer were added as additives to complement the nutriments of the soil samples before total petroleum hydrocarbon quantification and remediation procedures. Factorial design was applied to vary the input parameters such as pH, mass of substrate, moisture content and turning times of land farming so to ascertain the optimal conditions for the procedure. The result revealed that the in-situ TPH value was 5,000 mg/kg on the average and after 90 days of treatment, the turning rate, pH, moisture content and mass of substrate had 82.79%, 4.36%, 0.48% and 0.046% contributions respectively to land farming treatment. Numerical optimization techniques applied in the optimum point for land farming input parameters to achieve predicted maximum removal of 98.60% were evaluated as pH, mass of substrate, moisture content and turning rate to be 6.01, 1 kg, 10% and 5 times in a week respectively. TPH removed at this optimum point was 97.83% reducing from 5,000 mg/kg to 645.9 kg/mg. The high coefficient of determination (r2 = 0.9865) as observed in the closeness of predicted and experimental values reflects the reliability of the model and hence, land farming practice with close attention on turning rate as revealed by this study, is recommended for TPH contaminated soil remediation.
Exosomes Derived from miR-301a-3p-Modified Adipose-derived Mesenchymal Stem Cells Reverse Hypoxia-induced Erectile Dysfunction in Rat Models
Authors: Liang Li , Zheng Dachao , Lu Chao , Xi Qinghong , Bao Hua , Li Wengfeng , Gu Yufei , Mao Yuanshen , Xu Bin , Gu Xin
Published: August 2020

Erectile dysfunction (ED) has often been observed in patients with obstructive sleep apnea (OSA). Research on adipose-derived stem cells (ADSCs)-derived exosomes has shown that they have significant therapeutic effects in many diseases including ED. In this study, ED was induced in Sprague Dawley (SD) rats using chronic intermittent hypoxia (CIH) exposure. CIH-mediated influences were then measured in the corpus cavernous smooth muscle cells (CCSMCs). Results showed that miR-301a-3p-enriched exosomes treatment significantly recovered erectile function in rats and CCSMCs by promoting autophagy and inhibiting apoptosis. The treatment also significantly recovered the level of alpha Smooth Muscle Actin (α-SMA) in rats and CCSMCs. Bioinformatics predicted that Phosphatase and tensin homolog (PTEN) and Toll-like receptor 4 (TLR4) might be targets of miR-301a-3p. Our results indicate that PTEN-overexpression vectors or TLR4-overexpression vectors reverse the therapeutic effects achieved by miR-301a-3p in CCSMCs indicating that PTEN/HIF-1α and TLR4 signaling pathways play key roles in the progression of ED. The findings in this study suggest that miR-301a-3p should be considered as a new therapeutic target for treating ED associated with OSA.
Molecular Remission at T cell level in Patients with Rheumatoid Arthritis
Authors: Inamo Jun , Suzuki Katsuya , Takeshita Masaru , Kondo Yasushi , Okuzono Yuumi , Koga Keiko , Kassai Yoshiaki , Takiguchi Maiko , Kurisu Rina , Morita Rimpei , Yoshimura Akihiko , Takeuchi Tsutomu
Published: August 2020

Background: While numerous disease-modifying anti-rheumatic drugs (DMARDs) has brought about a dramatic paradigm shift in the management of rheumatoid arthritis (RA), unmet needs remain such as small proportion of achievement drug-free patients. The aim of this study is to explore key molecules for remission at T cell level, which is known to involve the pathogenesis of RA deeply, and investigate disease course of patients who achieved molecular remission (MR). Methods: We enrolled a total of 46 patients with RA and 10 healthy controls (HCs). We performed gene expression profiling and selected remission signature genes in CD4 + T cells and CD8 + T cells of patients with RA using machine learning methods. In addition, we investigated the benefit of achieving MR on disease control thereafter. Results: We identified 9 and 23 genes that were associated with clinical remission in CD4 + and CD8 + T cells. Principal component analysis (PCA) demonstrated expression profiling of them was similar with HCs. As to remission signature genes in CD4 + T cells, PCA result was reproduced using validation cohort, indicating their robustness. There was a trend towards better disease control during 12 months follow-up in patients treated with tocilizumab in deep MR than those in non-deep MR, although it was not significant. Conclusion: We identified robust genes which represented remission status in CD4 + T cells using machine learning techniques. The current study will promote our understandings about molecular mechanism to achieve deep remission in the management of RA.Trial registration: Not required.
C deletion of the SLC39A8 gene polymorphism (rs74650330) increases the risk of coronary artery disease in individuals with low LDL cholesterol levels
Authors: Zhang Jun , Yu Yang , Pan Lili , Yu Tianhong , Luo Guanghua
Published: August 2020

Background: The genetic variant of SLC39A8 is associated with several cardiovascular disease risk factors, including body mass index, systolic blood pressure (SBP), diastolic blood pressure (DBP), N-terminal pro-B-type natriuretic peptide (NT-proBNP) and high-density lipoprotein cholesterol (HDL-C) levels. The present study aimed to investigate the association between the SLC39A8 SNPs rs13107325 and rs74650330 and CAD in the Han population in Jiangsu (China).Methods: Genotyping of these SNPs was performed in 258 patients with CAD and 170 healthy controls using the base-quenched probe technique. The association between rs74650330 and blood lipid and glucose profiles was investigated.Results: The rs13107325 polymorphism was not found in the 428 Chinese individuals enrolled in the current study. For rs74650330, individuals harboring the C allele had significantly higher HDL levels than those without this allele in the control group (P=0.039), while the opposite was true for LDL-C levels (P=0.046). Further analysis indicated that when LDL-C levels were lower than 2.365 mmol/L, subjects with C/del and del/del had a 7.293-fold increased risk of CAD compared with that of controls without the mutation (odds ratio: 7.293; 95% confidence interval: 0.953-55.79).Conclusions: The results obtained in the current study suggested that the rs74650330 polymorphism is associated with reduced HDL-C and elevated LDL-C levels in healthy individuals. When LDL-C levels were lower than 2.365 mmol/L, the C/del genotype significantly increased the risk of CAD in the Chinese Han population.
Generation of induced tumor-suppressing cells from osteoblasts and mesenchymal stem cells
Authors: Sun Xun , Zha Rongrong , Li Kexin , Liu Shengzhi , Fan Yao , Wu Di , Hase Misato , Robling Alexander , Li Bai-Yan , Yokota Hiroki
Published: August 2020

Background

The current treatment for breast cancer-associated bone metastasis is palliative, and there is an unmet need to develop an effective regimen. In this study, we examined the possibility of conditioned medium (CM)-based therapy with bone-forming osteoblasts and their precursors, mesenchymal stem cells (MSCs).
Methods

Tumor cell lines, human breast cancer tissues, and a mouse model of mammary tumors and bone metastasis were employed to examine the tumor-suppressing effect of osteoblast- and MSC-derived CM. Tumor-driven osteolysis was evaluated using micro CT imaging and histology, and tumorigenic signaling was analyzed with an antibody-based protein array and Western blotting in response to the overexpression and RNA silencing of the key regulatory factors.
Results

Osteoblast- and MSC-derived CM without any overexpression did not alter the progression of mammary tumor cells. However, overexpressing Lrp5 and Snail converted them into induced tumor-suppressing (iTS) cells. In the mouse model, the systemic administration of iTS cell-derived CM reduced the weight of mammary tumors and suppressed tumor-induced osteolysis. The same outcome was observed with the overexpression of p53, a tumor-suppressing gene. In Lrp5-overexpressing CM, cytokines such as CXCL2 and LIF were downregulated, while p53 and Trail, an apoptosis-inducing factor, were upregulated. The overexpression of p53 in MSCs also reduced CXCL2 and LIF with an elevation of Trail. Notably, iTS cells were generated from mammary tumor cells by the overexpression of Lrp5 or Snail. We postulate that natural selection via cell competitions might be involved in the anti-tumor capability of iTS cells against tumor cells.
Conclusions

This study demonstrates that osteoblasts and MSCs can be converted into iTS cells by overexpressing the selected genes, and p53 is involved in the action of iTS cells. An iTS CM-based therapy may provide a novel option to treat primary and metastatic breast cancers.
PDIA3 Reduce Glioma-associated Macrophage/microglia Pro-tumor Activation trough IL-6-STAT3-PDIA3 Pathway.
Authors: Chiavari Marta , Ciotti Gabriella Maria Pia , Canonico Francesco , Altieri Fabio , Navarra Pierluigi , Lisi Lucia
Published: August 2020

Background: Glioblastoma (GB - grade IV glioma) is the most aggressive and common cancer of central nervous system with an overall survival of 14-16 months. The GB tumor microenvironment includes cells of the innate immune system identified as glioma-associated microglia/macrophages (GAMs). It is known that between GAMs and GB cells there is a double interaction, but the role of GAMs is still poorly characterized. The endoplasmic reticulum (ER) protein ERp57, also known as PDIA3, is a thiol oxidoreductase with main function related on glycoprotein folding in endoplasmic reticulum. However, PDIA3 shows different functions. In fact, the various subcellular localizations and binding partners of PDIA3 affect numerous physiological processes and diseases: different regulation and modulation of PDIA3 has been reported in multiple pathologies including neurodegenerative diseases and cancer. Methods: In the present work, we evaluated in both GB cells and microglia-macrophage cells the expression of PDIA3 using specimens collected after surgical from 18 GB patients. In addition, we studied in vitro microglia-glioma interaction to determine the role of PDIA3 in viability and the activation of both GB and microglia cells. The study was carried using PDIA3-silenced T98G cells and/or using a pharmacological inhibitor of PDIA3 activity (Punicalagin-PUN).Results: We initially investigated the role of the PDIA3 in GB survival by inquiring The Cancer Genome Atlas dataset. The results indicated that 352 out of 690 patients reported over-expression of PDIA3, which significantly correlated with a ~55% reduction of overall survival. Subsequently, for the first time, we investigated the PDIA3 expression in the tumor and the nearby parenchyma of 18 GB patients and our data showed a significant upregulation (15% vs 10%) of ERp57/PDIA3 in GAMs of tumor specimens respect the microglia present in parenchyma. In addition, we show that conditioned medium (CMs) obtained from both wild type T98G and PDIA3 silenced T98G induced an activation of microglia cells, but the PDIA3 silenced-T98G CMs significant limited the microglia pro-tumor activation probably through a IL-6-STAT3-PDIA3 dependent mechanism. Conclusion: Our data support the relevant role of PDIA3 expression in GB pathology and link the different activation of microglia to a mechanism a IL-6-STAT3-PDIA3 dependent.
CXCL11 Correlates with Anti-tumor Immune Microenvironment and Improved Prognosis in Colon Cancer
Authors: Cao Yingying , Zhang Youwei , Jiao Nanlin , Sun Tiantian , Ma Yanru , Zhang Xinyu , Hong Jie , CHEN HAOYAN
Published: August 2020

Background: CXCL11 has been considered to be responsible for tumor development, but the specific effect of CXCL11 in colon cancer was still obscure. Therefore, the prognostic value and immunological regulation effect of CXCL11 in colon cancer were evaluated in this study.Methods: Three independent datasets were used for mRNA-related analysis: one dataset from the Cancer Genome Atlas (TCGA, n=451) and two single-cell RNA sequencing (scRNA-seq) datasets from Gene Expression Omnibus (GEO): GSE146771 and GSE132465. In addition, the patient cohort (the Yijishan Hospital cohort, YJSHC, n=108) was utilized for cell infiltration-related analysis, accordingly. Both CXCL11 mRNA expression and CXCL11+ (CXCL11-producing) cells were assessed in colon cancer, whose effect on prognosis and immunological regulation was also studied. Results: High CXCL11 expression were associated with better prognosis in colon cancer, which was still significant even if clinicopathological factors were adjusted. Furthermore, CXCL11 positively correlated with anti-tumor cells infiltration, such as CD8+ T cells and natural killer cells. Meanwhile, CXCL11 correlated positively with several genes associated with DC, NK and T recruitment，and a gene set of cytotoxic genes. Notably, CXCL11 correlated positively with several immune checkpoint related genes including of PD-L1. Conclusions: CXCL11 contributed to anti-tumor immune microenvironment and could improve prognosis in patients with colon cancer. Especially, it’s a potential approach that inducible expression of CXCL11 by genetic and pharmacological interventions is able to improve prognosis and response to anti-PD-1 (programmed cell death protein-1) antibody treatment in colon cancer. However, it requires to be verified by further prospective investigations.
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

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

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

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

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

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

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

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

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 
Authors: Siempos Ilias , Xourgia Elena , Ntaidou Theodora K. , Zervakis Dimitris , Magira Eleni E. , Kotanidou Anastasia , Routsi Christina , Zakynthinos Spyros G.
Published: August 2020

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.