bluuebunny/crossref_metadata_2025_split huggingface dataset:
Umbilical cord as a long-term source of activatable mesenchymal stromal cells for immunomodulation
Authors: Selich Anton , Zimmermann Katharina , Tenspolde Michel , Dittrich-Breiholz Oliver , von Kaisenberg Constantin , Schambach Axel , Rothe Michael 
Published: September 2019

Abstract

Background
Mesenchymal stromal cells (MSCs) are used in over 800 clinical trials mainly due to their immune inhibitory activity. Umbilical cord (UC), the second leading source of clinically used MSCs, is usually cut in small tissue pieces. Subsequent cultivation leads to a continuous outgrowth of MSC explant monolayers (MSC-EMs) for months. Currently, the first MSC-EM culture takes approximately 2 weeks to grow out, which is then expanded and applied to patients. The initiating tissue pieces are then discarded. However, when UC pieces are transferred to new culture dishes, MSC-EMs continue to grow out. In case the functional integrity of these cells is maintained, later induced cultures could also be expanded and used for cell therapy. This would drastically increase the number of available cells for each patient. To test the functionality of MSC-EMs from early and late induction time points, we compared the first cultures to those initiated after 2 months by investigating their clonality and immunomodulatory capacity.

Methods
We analyzed the clonal composition of MSC-EM cultures by umbilical cord piece transduction using integrating lentiviral vectors harboring genetic barcodes assessed by high-throughput sequencing. We investigated the transcriptome of these cultures by microarrays. Finally, the secretome was analyzed by multiplexed ELISAs, in vitro assays, and in vivo in mice.

Results
DNA barcode analysis showed polyclonal MSC-EMs even after months of induction cycles. A transcriptome and secretome analyses of early and late MSC cultures showed only minor changes over time. However, upon activation with TNF-α and IFN-γ, cells from both induction time points produced a multitude of immunomodulatory cytokines. Interestingly, the later induced MSC-EMs produced higher amounts of cytokines. To test whether the different cytokine levels were in a therapeutically relevant range, we used conditioned medium (CM) in an in vitro MLR and an in vivo killing assay. CM from late induced MSC-EMs was at least as immune inhibitory as CM from early induced MSC-EMs.

Conclusion
Human umbilical cord maintains a microenvironment for the long-term induction of polyclonal and immune inhibitory active MSCs for months. Thus, our results would offer the possibility to drastically increase the number of therapeutically applicable MSCs for a substantial amount of patients.
miR-106b regulates the proliferation and differentiation of neural stem/progenitor cells through Tp53inp1-Tp53-Cdkn1a axis
Authors: Xia Xiaohuan , Lu Hongfang , Li Chunhong , Huang Yunlong , Wang Yi , Yang Xiaoyu , Zheng Jialin C. 
Published: September 2019

Abstract

Background
Recent studies suggested that miR-17~106 family was involved in the regulation of neural stem/progenitor cells (NPCs). However, distinct function of each family member was reported in regulating stem cells within and without the brain. Hence, to investigate the roles of individual miRNAs in miR-17~106 family and mechanisms underlying their effects on neurogenesis is important to extend our understanding in the CNS development.

Methods
Here, we examined the influence of miR-106a/b on the proliferation, differentiation, and survival of embryonic NPCs using specific mimics and inhibitor. The targets of miR-106a/b were identified from miRNA target prediction database and confirmed by luciferase assay. Specific siRNAs were utilized to erase the effects of miR-106a/b on the expression levels of target genes.

Results
A positive correlation was observed between the temporal reduction of miR-106a/b expression levels and the decline of NPC pools in vivo and in vitro. The perturbation of miR-106’s function approaches revealed that miR-106b, but not miR-106a, facilitated the maintenance of NPCs and repressed the generation of both neuronal and glial cells, without preference to a particular lineage. No effect was observed for miR-106a/b in NPCs’ survival. The influence of miR-106b on NPCs’ proliferation and differentiation is likely achieved by directly inhibiting the expression of Tp53inp1 and Cdkn1a, key components of Tp53inp1-Tp53-Cdkn1a axis.

Conclusion
Our study demonstrated a novel axis, miR-106b-Tp53inp1-Tp53-Cdkn1a, in regulating the proliferation and differentiation of NPCs.
Mechanical process prior to cryopreservation of lipoaspirates maintains extracellular matrix integrity and cell viability: evaluation of the retention and regenerative potential of cryopreserved fat-derived product after fat grafting
Authors: Feng Jingwei , Hu Wansheng , Fanai Mimi Lalrimawii , Zhu Shengqian , Wang Jing , Cai Junrong , Lu Feng 
Published: September 2019

Abstract

Background
Cryopreservation of fat grafts facilitates reinjection for later use. However, low temperature and thawing can disrupt tissues and cause lipid leakage, which raises safety concerns. Here, we compared the cryopreservation potential of stromal vascular fraction (SVF) gel processed from lipoaspirate with that of fat.

Methods
Human SVF gel and fat were cryopreserved at − 20 °C without cryoprotectant for 1 month. Fresh SVF gel and fat were used as controls. Tissue viability, adipose-derived stem cell (ASC) function, and the extracellular content were evaluated. At 3 months after transplanting the specimens to immunocompromised mice subcutaneously, the grafts were examined for retention, tissue engraftment, and inflammatory levels. The regenerative effect of cryopreserved SVF gel was evaluated in a murine ischemic wound healing model.

Results
At 1 month, the cell death rate in the SVF gel group was 36 ± 2%. The survived ASCs not only could be isolated via explant culture but also preserved colony-forming and differentiation. However, prolonged cryopreservation exacerbated apoptosis. Assessment of recovered tissues showed that the morphology, cell viability, and extracellular protein enrichment were better in SVF gel-preserved tissues than in frozen fat. At 3 months after lipotransfer, the retention ability of 1-month cryopreserved fat was 41.1 ± 9% compared to that of 1-month cryopreserved SVF gel. Immunostaining results showed that adipose tissue regeneration and integrity in the 1-month cryopreserved SVF gel group were superior to those of the cryopreserved fat group. The cryopreserved SVF gel also accelerated healing of the ischemic wound, compared with cryopreserved fat.

Conclusion
Cryopreserved SVF gel maintained tissue integrity and cell viability and resulted in a better long-term retention rate than that of cryopreserved fat. Cryopreserved SVF gel also showed superior regenerative potential and improved ischemic wound healing.
iPSC-derived MSC therapy induces immune tolerance and supports long-term graft survival in mouse orthotopic tracheal transplants
Authors: Khan Mohammad Afzal , Alanazi Fatimah , Ahmed Hala Abdalrahman , Shamma Talal , Kelly Kilian , Hammad Mohamed A. , Alawad Abdullah O. , Assiri Abdullah Mohammed , Broering Dieter Clemens 
Published: September 2019

AbstractBackgroundLung transplantation is a life-saving surgical replacement of diseased lungs in patients with end-stage respiratory malfunctions. Despite remarkable short-term recovery, long-term lung survival continues to face several major challenges, including chronic rejection and severe toxic side effects due to global immunosuppression. Stem cell-based immunotherapy has been recognized as a crucial immunoregulatory regimen in various preclinical and clinical studies. Despite initial therapeutic outcomes, conventional stem cells face key limitations. The novel Cymerus™ manufacturing facilitates production of a virtually limitless supply of consistent human induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells, which could play a key role in selective immunosuppression and graft repair during rejection.MethodsHere, we demonstrated the impact of iPSC-derived human MSCs on the development of immune tolerance and long-term graft survival in mouse orthotopic airway allografts. BALB/c → C57BL/6 allografts were reconstituted with iPSC-derived MSCs (2 million/transplant/at d0), and allografts were examined for regulatory T cells (Tregs), oxygenation, microvascular blood flow, airway epithelium, and collagen deposition during rejection.ResultsWe demonstrated that iPSC-derived MSC treatment leads to significant increases in hTSG-6 protein, followed by an upregulation of mouse Tregs and IL-5, IL-10, and IL-15 cytokines, which augments graft microvascular blood flow and oxygenation, and thereby maintained a healthy airway epithelium and prevented the subepithelial deposition of collagen at d90 post transplantation.ConclusionsCollectively, these data confirmed that iPSC-derived MSC-mediated immunosuppression has potential to establish immune tolerance and rescue allograft from sustained hypoxic/ischemic phase, and subsequently limits long-term airway epithelial injury and collagen progression, which therapeutically warrant a study of Cymerus iPSC-derived MSCs as a potential management option for immunosuppression in transplant recipients.
Haemophilus parasuis VtaA2 is involved in adhesion to extracellular proteins
Authors: Costa-Hurtado Mar , Garcia-Rodriguez Laura , Lopez-Serrano Sergi , Aragon Virginia 
Published: September 2019

Abstract
Haemophilus parasuis is part of the microbiota of the upper respiratory tract in swine. However, virulent strains can cause a systemic disease known as Glässer’s disease. Several virulence factors have been described in H. parasuis including the virulence-associated trimeric autotransporters (VtaAs). VtaA2 is up-regulated during infection and is only found in virulent strains. In order to determine its biological function, the vtaA2 gene was cloned with its native promotor region in pACYC184, and the transformed Escherichia coli was used to perform functional in vitro assays. VtaA2 was found to have a role in attachment to plastic, mucin, BSA, fibronectin and collagen. As other VtaAs from H. parasuis, the passenger domain of VtaA2 contains collagen domains. In order to examine the contribution of the collagen repeats to VtaA2 function, a recombinant vtaA2 without the central collagen domains was obtained and named vtaA2OL. VtaA2OL showed similar capacity than VtaA2 to adhere to plastic, mucin, BSA, fibronectin and plasma but a reduced capacity to adhere to collagen, suggesting that the collagen domains of VtaA2 are involved in collagen attachment. No function in cell adhesion and invasion to epithelial alveolar cell line A549 or unspecific binding to primary alveolar macrophages was found. Likewise VtaA2 had no role in serum or phagocytosis resistance. We propose that VtaA2 mediates adherence to the host by binding to the mucin, found in the upper respiratory tract mucus, and to the extracellular matrix proteins, present in the connective tissue of systemic sites, such as the serosa.
Urothelial carcinoma associated 1 promotes trophoblast invasion by regulating MMP9
Authors: Shao Hongfang , Jin Feng , Hu Jiangshan , Zhu Zhangying , Tian Fuju , Tao Minfang , Teng Yincheng 
Published: September 2019

Abstract

Background
The long non-coding RNA UCA1 is reportedly increased in several human tumors and critical for the cell migration, invasion, or proliferation of several cancer cells. However, the potential roles of UCA1 in trophoblasts at early pregnancy still poorly understood. Here, we sought to unravel the roles of UCA1 in the occurrence of the recurrent miscarriage (RM) disorders.

Results
The knockdown of UCA1 in human HTR-8 trophoblast cell line reduced their cell proliferative and invasive ability. Conversely, the UCA1 overexpression promoted the cell proliferation and invasion of HTR-8 cells. Quantitative RT-PCR screening revealed that UCA1 overexpression significantly enhanced MMP9, but not MMP2, mRNA expression in trophoblast cells. The overexpression of UCA1 also promoted trophoblast invasion by upregulating MMP9 expression and activity both in vitro and ex vivo. Consistently, UCA1 and MMP9 mRNA expression level was notably reduced in placental villi derived from patients with RM diseases.

Conclusion
This study revealed that UCA1 is critical for the regulation of invasive ability in trophoblasts. The abnormal UCA1/MMP9 pathway might result in the impaired trophoblast activities and lead to the development of RM. Our data may also provide a novel angle for the treatment in RM patients.
Nephrotoxic drug burden among 1001 critically ill patients: impact on acute kidney injury
Authors:   Clinical research in intensive care and sepsis-Trial group for global evaluation and research in sepsis (CRICS-TRIGGERSEP network), Ehrmann Stephan , Helms Julie , Joret Aurélie , Martin-Lefevre Laurent , Quenot Jean-Pierre , Herbrecht Jean-Etienne , Benzekri-Lefevre Dalila , Robert René , Desachy Arnaud , Bellec Fréderic , Plantefeve Gaëtan , Bretagnol Anne , Dargent Auguste , Lacherade Jean-Claude , Meziani Ferhat , Giraudeau Bruno , Tavernier Elsa , Dequin Pierre-François 
Published: September 2019

Abstract

Background
Nephrotoxic drug prescription may contribute to acute kidney injury (AKI) occurrence and worsening among critically ill patients and thus to associated morbidity and mortality. The objectives of this study were to describe nephrotoxic drug prescription in a large intensive-care unit cohort and, through a case–control study nested in the prospective cohort, to evaluate the link of nephrotoxic prescription burden with AKI.

Results
Six hundred and seventeen patients (62%) received at least one nephrotoxic drug, among which 303 (30%) received two or more. AKI was observed in 609 patients (61%). A total of 351 patients were considered as cases developing or worsening AKI a given index day during the first week in the intensive-care unit. Three hundred and twenty-seven pairs of cases and controls (patients not developing or worsening AKI during the first week in the intensive-care unit, alive the case index day) matched on age, chronic kidney disease, and simplified acute physiology score 2 were analyzed. The nephrotoxic burden prior to the index day was measured in drug.days: each drug and each day of therapy increasing the burden by 1 drug.day. This represents a semi-quantitative evaluation of drug exposure, potentially easy to implement by clinicians. Nephrotoxic burden was significantly higher among cases than controls: odds ratio 1.20 and 95% confidence interval 1.04–1.38. Sensitivity analysis showed that this association between nephrotoxic drug prescription in the intensive-care unit and AKI was predominant among the patients with lower severity of disease (simplified acute physiology score 2 below 48).

Conclusions
The frequently observed prescription of nephrotoxic drugs to critically ill patients may be evaluated semi-quantitatively through computing drug.day nephrotoxic burden, an index significantly associated with subsequent AKI occurrence, and worsening among patients with lower severity of disease.
A new viscosity-type iteration for a finite family of split variational inclusion and fixed point problems between Hilbert and Banach spaces
Authors: Izuchukwu C. , Isiogugu F. O. , Okeke C. C. 
Published: September 2019

Abstract
In this paper, we introduce a new viscosity-type iteration process for approximating a common solution of a finite family of split variational inclusion problem and fixed point problem. We prove that the proposed algorithm converges strongly to a common solution of a finite family of split variational inclusion problems and fixed point problem for a finite family of type-one demicontractive mappings between a Hilbert space and a Banach space. Furthermore, we applied our results to study a finite family of split convex minimization problems, and also considered a numerical experiment of our results to further illustrate its applicability. Our results extend and improve the results of Byrne et al. (J. Nonlinear Convex Anal. 13:759–775, 2012), Kazmi and Rizvi (Optim. Lett. 8(3):1113–1124, 2014), Moudafi (J. Optim. Theory Appl. 150:275–283, 2011), Shehu and Ogbuisi (Rev. R. Acad. Cienc. Exactas Fís. Nat., Ser. A Mat. 110(2):503–518, 2016), Takahashi and Yao (Fixed Point Theory Appl. 2015:87, 2015), Chidume and Ezeora (Fixed Point Theory Appl. 2014:111, 2014), and a host of other important results in this direction.
Fractional order ecological system for complexities of interacting species with harvesting threshold in imprecise environment
Authors: Khan Najeeb Alam , Razzaq Oyoon Abdul , Mondal Sankar Parsad , Rubbab Qammar 
Published: September 2019

Abstract
The key objective of this paper is to study the imprecise biological complexities in the interaction of two species pertaining to harvesting threshold. It is explained by taking the prey–predator model with imprecise biological parameters and fractional order generalized Hukuhara (fgH) differentiability. In this vain, different possible systems of the model are constructed, according to the increasing and decreasing behavior of population growth. Feasibility and stability analyses of equilibrium points of the stated models are also discussed by means of variational matrix with Routh–Hurwitz conditions. In addition, the numerical elaborations are carried out by taking parametric expansion of fuzzy fractional Laplace transform (FFLT). This significantly helps the researchers in using a novel approach to analyze the constant solutions of the dynamical systems in the presence of fractional index. This would allow the avoidance of any intricacy that occurs while solving fractional order derivatives. Furthermore, this attempt also provides numerical and pictorial results, obtained through some well-known methods, namely fifth-forth Runge–Kutta method (FFRK), Grunwald–Letnikov’s definition (GL) and Adams–Bashforth method (ABM) that are deemed appropriate to scrutinize the dynamics of the system of equations.
Amplified fragment length polymorphism and whole genome sequencing: a comparison of methods in the investigation of a nosocomial outbreak with vancomycin resistant enterococci
Authors: Janes Victoria A. , Notermans Daan W. , Spijkerman Ingrid J.B. , Visser Caroline E. , Jakobs Marja E. , van Houdt Robin , Willems Rob J.L. , de Jong Menno D. , Schultsz Constance , Matamoros Sébastien 
Published: September 2019

Abstract

Background
Recognition of nosocomial outbreaks with antimicrobial resistant (AMR) pathogens and appropriate infection prevention measures are essential to limit the consequences of AMR pathogens to patients in hospitals. Because unrelated, but genetically similar AMR pathogens may circulate simultaneously, rapid high-resolution molecular typing methods are needed for outbreak management. We compared amplified fragment length polymorphism (AFLP) and whole genome sequencing (WGS) during a nosocomial outbreak of vancomycin-resistant Enterococcus faecium (VRE) that spanned 5 months.

Methods
Hierarchical clustering of AFLP profiles was performed using unweighted pair-grouping and similarity coefficients were calculated with Pearson correlation. For WGS-analysis, core single nucleotide polymorphisms (SNPs) were used to calculate the pairwise distance between isolates, construct a maximum likelihood phylogeny and establish a cut-off for relatedness of epidemiologically linked VRE isolates. SNP-variations in the vanB gene cluster were compared to increase the comparative resolution. Technical replicates of 2 isolates were sequenced to determine the number of core-SNPs derived from random sequencing errors.

Results
Of the 721 patients screened for VRE carriage, AFLP assigned isolates of 22 patients to the outbreak cluster. According to WGS, all 22 isolates belonged to ST117 but only 21 grouped in a tight phylogenetic cluster and carried vanB resistance gene clusters. Sequencing of technical replicates showed that 4–5 core-SNPs were derived by random sequencing errors. The cut-off for relatedness of epidemiologically linked VRE isolates was established at ≤7 core-SNPs. The discrepant isolate was separated from the index isolate by 61 core-SNPs and the vanB gene cluster was absent. In AFLP analysis this discrepant isolate was indistinguishable from the other outbreak isolates, forming a cluster with 92% similarity (cut-off for identical isolates ≥90%). The inclusion of the discrepant isolate in the outbreak resulted in the screening of 250 patients and quarantining of an entire ward.

Conclusion
AFLP was a rapid and affordable screening tool for characterising hospital VRE outbreaks. For in-depth understanding of the outbreak WGS was needed. Compared to AFLP, WGS provided higher resolution typing of VRE isolates with implications for outbreak management.
Simultaneous determination of various platycosides in Four Platycodon grandiflorum cultivars by UPLC-QTOF/MS
Authors: Lee Dae Young , Choi Bo-Ram , Lee Jae Won , Um Yurry , Yoon Dahye , Kim Hyoung-Geun , Lee Young-Seob , Kim Geum-Soog , Lee Youn-Hyung , Baek Nam-In 
Published: September 2019

Abstract
In Platycodi Radix (root of Platycodon grandiflorum), there are a number of platycosides that consist of a pentacyclic triterpenoid aglycone and two sugar moieties. Due to the pharmacological activities of platycosides, it is critical to assess their contents in PR, and develop an effective method to profile various platycosides is required. In this study, an analytical method based on ultra performance liquid chromatography coupled with quadrupole time-of-flight/mass spectrometry (UPLC-QTOF/MS) with an in-house library was developed and applied to profile various platycosides from four different Platycodi Radix cultivars. As a result, platycosides, including six isomeric pairs, were successfully analyzed in the PRs. In the principal component analysis, several platycosides were represented as main variables to differentiate the four Platycodi Radix cultivars. Their different levels of platycosides were also represented by relative quantification. Finally, this study indicated the proposed method based on the UPLC-QTOF/MS can be an effective tool for identifying the detail characterization of various platycosides in the Platycodi Radix.