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BAS-011 exam Dumps Source : IBM SPSS Statistics level 1 v2
Test Code : BAS-011
Test name : IBM SPSS Statistics level 1 v2
Vendor name : IBM
: 55 true Questions
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IBM IBM SPSS Statistics Level
The IBM Hybrid Cloud team is back at it with yet another win for design. I’m excited to announce that their design crew has been awarded the 2018 pink Dot: conversation Design Award for IBM SPSS statistics within the Interface Design category. This award is a continuation of the design achievements they fill seen this previous 12 months, together with the A’Design Awards, IF Design Awards, and others. i am overjoyed to peer the complicated toil of their designers and IBM Design continue to shine and result a change in business utility.
First developed within the 1990’s, the red Dot Award has been the revered overseas seal of wonderful design pleasant. Designers, businesses, and organizations from forty five diverse countries took half in this year’s competition, totaling over eight,600 entries that underwent a 24 member jury.
“All those that evolution during the complicated adjudication system to garner a pink Dot fill every rationale to breathe pleased with themselves, as the jury promises their award best to creations of excessive design quality. This makes me totality of the greater delighted to congratulate the laureates essentially on their richly deserved success.” — Professor Dr. Peter Zec, founder and CEO of the crimson Dot Award
Receiving this award turned into totally exciting for their team and we're honored to breathe among the winners. here's a tall fulfillment for their designers who worked on this product, and that they faced an intriguing and difficult adventure in engaged on this product.
what is IBM SPSS?
IBM SPSS information is a powerful facts analysis implement that is likely one of the most accepted records functions. considering the fact that its inception in 1968, SPSS statistics has been revamped and redeveloped distinct instances. Now the design team at IBM has taken on the assignment of developing a very clean consumer journey.
during this newest remodel of IBM SPSS statistics, they implemented design thinking ideas by working closely with their users and making confident this modernized edition of SPSS data aligns with their wants. Their foremost purpose turned into to create a magnificent device that is not simplest effortless and intuitive to result consume of, but that their users can enjoy.
Our group and Design approach
The IBM SPSS design crew is fraction of the IBM Design Studios in Boeblingen, Germany. The crew consists of a diverse community, with many contributors originating from distinctive countries and cultures. Some participants of the group had some background with statistics while others had been working in this container for the first time.
Following the concepts of IBM Design thinking (study > reflect > Make), their group implemented a redecorate that brings a stronger focus on users for SPSS facts. The design team carried out intensive analysis on the user foundation of SPSS information with the aim to espy how the utility can more advantageous meet their wants. The current person foundation degrees from less skilled users comparable to students to more professional clients corresponding to statistics scientists or business professionals. A key insight from the group’s research become that less skilled users had been intimidated each with the aid of the mathematics toil and the complexity of the application.
the brand fresh designs concentrated on simplifying workflows, reducing the common complexity of the UI and interactions, and proposing newcomers an light on-boarding to information and to the product. a further faultfinding office within the remodel changed into a working towards e-book led by using a personality named Simon, who serves as an in-utility e-book, assisting novice users recollect diverse features and achieve their dreams sooner.
The group faced some entertaining challenges in redesigning a product of such complexity, and one which has additionally been round for therefore decades. a great success of the designers changed into making the product purchasable and attractive to fresh users without alienating decade-lengthy, experienced users.
a view Into the Future
The preview version of their fresh IBM SPSS facts event was launched in March 2018, and made attainable to the public as a crucible on the IBM consider conference is Las Vegas, and for the reason that June 26 , the fresh UI is commonly available to totality SPSS records subscribers. This preview is just the initial step, providing essentially the most used statistical analyses, and basic capabilities for data preparation, for presentation and for reporting outcomes. Over here months the team should breathe working so as to add extra aspects and capabilities with a purpose to meet experience needs of totality of their person groups.
no longer just Updating — Redesigning
i'm so thrilled to espy a further Hybrid Cloud design group receive an international award for his or her work. IBM SPSS data is yet another illustration of how design is making an primary change within the success of their products. As they continue to result consume of design to create greater relatable and efficacious products, they are able to provide their users the experiences that they want and want. I’m delighted and supercilious to monitor the incompatibility that their design group is making in the world of enterprise application, and i can’t wait to view how they continue to fill an repercussion on the lives of their users.
Design supervisor: Caroline law
Design Leads: Dirk Willuhn and Eva Cochet-Weinandt
Design crew: Christian Fritsche, Dimitri Hoffmann, Jaehee (Chloe) Lee, Oleksandr Sabov, Stephan Feger
because of these contributing designers: Katrin Ellice Heintze, Leila Johannesen, Marion Bruells, Phil Brucker, Robin Auer, Sammy Schuckert, Stefan Schwarz
Design interns: Mengzhu Deng, Nathalie Mader, Ting-Hao (Howard) Huang, Vanessa Ng
Two of IBM’s most prevalent evaluation items, the Cognos company Intelligence and the SPSS predictive analytics equipment, are headed for the cloud, the latest in an ongoing shove via IBM to port its tall utility portfolio to the cloud.
getting access to one of these utility from a hosted ambiance, rather than purchasing the package outright, gives a number of advantages to customers.
“We control the infrastructure, and this lets you scale more effectively and regain totality started with less upfront funding,” said Eric Sall, IBM vp of global analytics marketing.
IBM introduced these additions to its cloud functions, as well as a few fresh offerings, at its perception person conference for information analytics, held this week in Las Vegas.
by using 2016, 25 percent of latest business evaluation deployments might breathe accomplished in the cloud, according to Gartner.
Analytics may assist agencies in lots of techniques, based on IBM. It could provide extra insight within the paying for habits of shoppers, as well as insight into how smartly its personal operations are performing. It might aid occupy faith of methods from assaults and makes an attempt at fraud, as well as assure that company departments are assembly compliance requirements.
the fresh on-line version of Cognos, IBM Cognos business Intelligence on Cloud, can at present breathe demonstrated in a preview mode. IBM plans to proffer Cognos as a full commercial carrier early subsequent year. clients can sprint Cognos against data they preserve in the IBM cloud, or towards information they shop on premises.
A full industrial edition of the online IBM SPSS Modeler will breathe obtainable inside 30 days. This equipment will embrace totality of the SPSS components for facts based predictive modeling, comparable to a modeler server, analytics determination administration software and a information server.
earlier this yr, IBM pledged to proffer a edifying deal of its utility portfolio as cloud services, many via its Bluemix set of platform capabilities.
apart from Cognos and SPSS, IBM additionally unveiled a pair of fresh and up to date offerings on the conference.
One fresh provider, DataWorks, offers a number of ideas for refining and cleaning information so it is ready for analysis. The company has launched a cloud-primarily based data warehousing carrier, referred to as dashDB. a fresh Watson-based mostly carrier, called Watson Explorer, provides a mode for clients to question natural language questions on varied units of interior statistics.
To touch upon this text and other PCWorld content material, consult with their facebook page or their Twitter feed.
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Obesity has become a worldwide problem and a major risk factor for diabetes, infertility, and cardiovascular disease (Hu et al., 2016; Liang et al., 2017; Ferramosca et al., 2016; Supriya et al., 2018). It is a chronic metabolic disease characterized by abnormal stout distribution or excessive lipid accumulation (Yang et al., 2012). White adipose tissue (WAT), the central metabolic organ that regulates the energy homeostasis of the cadaver (Choe et al., 2016), is closely associated with the happening of obesity and related complications (Hajer, Van Haeften & Visseren, 2008).
Noncoding RNA (ncRNA) is a universal term for totality functional RNAs that are not translated into proteins. As biological mediators, they can participate in the regulation of gene expression through epigenetic modification, transcriptional regulation and post-transcriptional regulation (Tang, Chen & Zhou, 2018). With the evolution of sequencing technology, a variety of fresh non-coding RNAs fill been discovered, and their role in gene regulatory networks and regulation of endothelial cell office and metabolism is becoming better understood (Santulli, 2015, 2016; Liu et al., 2018). Among them, circular RNA (circRNA) and long non-coding RNA (lncRNA) play roles in regulating beta cell function, influencing transformation of adipose tissue and its energy metabolism and making them a promising target for anti-obesity therapy (Kaur, Mirza & Pociot, 2018; Li et al., 2018; Zhu et al., 2019).
Salvianolic acid B (Sal B) is a water-soluble component extracted from the traditional Chinese medicinal plant Salvia miltiorrhiza (Family Labiatae; referred to herein as S. miltiorrhiza) (Wang et al., 2014). In recent years, studies fill shown that Sal B can reduce obesity and obesity-related metabolic disorders (Chien et al., 2016; Pan et al., 2018). Their toil has shown that Sal B can ameliorate glycolipid metabolism and reduce cadaver weight in obese mice induced by towering stout diet (HFD) (Zhao et al., 2017). However, it is not known whether anti-obesity activity of Sal B is related to the regulation of non-coding RNA expression. In this study, they aimed to investigate the result of Sal B on the expression of lncRNA and circRNA in epididymis white adipose tissue (EP) of obese mice induced by a HFD. The anti-obesity target of Sal B was screened using functional studies of differentially expressed lncRNA and mRNA.
Materials and Methods
All the study protocol was approved by the Animal faith and Management Committee of the Beijing University of Chinese Medicine, and totality animal manipulations were according to the guidelines of the Animal faith Committee.
Mice experiment and tissue extraction
Male C57BL/6J mice provided by SPF (Beijing) Biotechnology Co. Ltd. (Certificate NO. SCXK (Jing) 2016-0002) were used in this study. After 1-week acclimation, the mice were fed a HFD (60% fat) for 12 weeks to induce obesity (average cadaver weight: HFD > (1 + 20%) normal, n = 10). After that, obesity mice were randomly divided into EP-S and EP-M groups (n = 5). Sal B intervention group (EP-S) and model (EP-M) groups were respectively administered Sal B (75 mg/kg cadaver weight/day) or vehicle (an equivalent volume of water) by oral gavage daily for 8 weeks. At the discontinue of the study, blood samples and EPs were collected from totality the mice sacrificed by cervical dislocation, and immediately frozen in liquid nitrogen and stored at −80 °C for subsequent analysis.
Measurement of blood lipid profiles and cadaver stout mass
The concentrations of serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL), and high-density lipoprotein cholesterol (HDL) were determined using chemistry reagent kits (Nanjing Jiancheng Biology Engineering Institute, Nanjing, China) and an automated biochemical analyzer (Hitachi, Tokyo, Japan). cadaver composition and total stout mass was measured by magnetic resonance imaging (EchoMRI-100 for mice; reflect Medical System, Houston, TX, USA) after the Sal B administration (weeks 8).
RNA isolation and RNA-seq analysis
Total RNAs from tissues were extracted using the Trizol reagent and purified using RiboZero Magnetic Gold Kit according to the manufacturer’s instructions. RNA sequencing libraries were generated using the KAPA Stranded RNA-Seq Library Prep Kit. The constructed cDNA libraries were qualified by Agilent 2100 Bioanalyzer, quantified by qPCR, and sequenced on an Illumina Hiseq 4000.
Functional enrichment analysis
Using Gene Ontology (GO) database (http://www.geneontology.org). They analysis the travel enrichment of the differentially expressed mRNAs and their functions, based on three aspects: biological processes (BP), cellular components (CC), and molecular functions (MF). The log 10 values (p-value) denote enrichment scores and picture the significance of the travel term enrichment among the differentially expressed genes. KEGG pathway analysis revealed pathway clusters covering the differentially expressed genes, and the log 10 values (p-value) denote the enrichment score and picture the significance of the pathway correlations. In addition, Gene Set Enrichment Analysis (GSEA) is used to compensate for the shortcomings of individual genes in analysis.
Correlation and co-expression analysis of mRNA and lncRNA
Based on the inter-regulatory association between differentially expressed genes in the EP between Sal B treatment group and obesity group, the lncRNA–mRNA regulatory network was constructed using Cytoscape v2.8.2 software (http://www.cytoscape.org/).
Quantitative true time-PCR
Quantitative true time (qRT)-PCR was performed to verify the results of RNA-seq. Total RNAs were isolated from samples using the Trizol reagent, then reverse-transcribed into cDNA according to the manufacturers’ instruction. The transcriptase reactions contained 1.5 μg RNA, 0.5 μg/μl random primers (N9, 1 μl), 2.5 mM dNTPs amalgamate (1.6 μl), 5× first-strand buffer (4.0 μl), 0.1M dithiothreitol (1 μl), RNase inhibitor (0.3 μl), and Superscript III RT (0.2 μl). Sybr Green-based qPCR was performed using SYBR Premix ExTaq. totality data were normalized to data for ARBP to calculate relative mRNA concentrations. The primers used in this study are shown in Table 1.
Primers for quantitative PCR analysis.
F: TTTGGGCATCACCACGAAAAR: GGACACCCTCCAGAATTTTC
F: TGAGCTCTTAAGAGTCACCAAGR: CTTGTTCTGATGTTGCATGCTC
F: CAAGATGCGCATTAAGGAGATCR: CTTGAGCAGCTTCTTCTTC
F: TCACACTCTCTTTGGTTTTR: CCAGTTGAAGCACAAATCTGAA
F: AGACCTCACGGTGCCAAATR: CTTTCTTTCTTAACGTCCACAGG
F: CAGTTCATGAAAGAAGCTGGTCR: CGAGCATGGAAGTATTTGTCTG
F: GGCAGGCATGACTAAATG 3R: CAGGGTTGATTAGCAGTGTC
The statistical differences were analyzed using the SPSS (version 20.0; IBM SPSS Statistics, Chicago, IL, USA) by independent-samples t-test. totality data were shown as the means ± SD. p-values < 0.05 were regarded as statistically significant.
Effects of Sal B on cadaver stout mass and serum lipid profiles of obese mice induced by HFD
After 8 weeks of Sal B intervention, the cadaver weights, TG, TC, HDL, LDL, and cadaver stout mass of experimental mice in the two groups were measured. The cadaver weight, TG, TC, HDL, LDL, and cadaver stout mass in the obesity group was significantly higher than that in Sal B treatment group (p < 0.05) (Table 2). These results attest that Sal B can reduce the cadaver weight and stout mass as well as obviate dyslipidemia caused by HFD feeding.
Effects of Sal B on cadaver stout mass and serum lipid profiles of obese mice induced by HFD.
Obesity model group
Sal B treatment group
0.984 ± 0.106
0.777 ± 0.069*
6.229 ± 0.483
5.011 ± 0.391*
0.425 ± 0.021
0.317 ± 0.029*
1.213 ± 0.191
1.876 ± 0.105*
0.431 ± 0.011
0.356 ± 0.015*
Effects of Sal B on mRNAs and circRNAs expression in EP of obese mice induced by HFD
In total, 15,184 mRNAs were identified, of which 132 differentially expressed (DEmRNAs). In the EP-S group, 24 were up-regulated and 108 were down-regulated (Figs. 1A and 1C; Table S1). Compared with EP-M, there were 19 differentially expressed circRNAs in the EP-S, of which nine were up-regulated and 10 were down-regulated (Figs. 1B and 1D; Table S2). Among DEmRNAs, the up-regulated expression of Wbscr27 was the highest, with a fold change of 2.053, while C1rb was the most down-regulated, with a fold change of 0.318. In addition, some mRNAs that fill been shown to play a role in stout metabolism, such as Sfrp5, Adig, and Saa3 were moreover differentially expressed.
Figure 1: Analysis of DEmRNAs (A, C) and DEcircRNAs (B, D).
Hierarchical clustering. Each row represents an mRNA and each column represents a sample. Green and red picture down-and up-regulated mRNAs or circRNAs, respectively. Genes in the volcano-Plot above the green parallel line (p < 0.05) and outside the two longitudinal green lines indicated DEmRNAs and DEcircRNAs between the two compared samples.
Effects of Sal B on the expression and office of lncRNAs in EP induced by HFD in obese mice
We establish 234 differentially expressed lncRNAs (DE lncRNAs), including 87 up-regulated and 147 down-regulated in the experimental group (Table S3). Based on this, they performed a GSEA functional analysis of the DElncRNAs, and establish that the up-regulated expression of lncRNAs are mainly involved in brown adipocyte differentiation, steroid biosynthesis, lipid transport, and lipid metabolism, while the down-regulated expression of lncRNAs are associated with the immune process and inflammatory responses (Fig. 2). After classifying the DElncRNAs, 179 were establish to breathe exon sense overlapping, 11 were intergenic, 17 were intron sense overlapping, 16 were antisense, and 11 were bidirectional.
Figure 2: GSEA Cluster Heat Map of top 10 DElncRNAs, in up-regulation and down-regulation, respectively.
(A) Biological process; (B) cellular components; (C) molecular functions, and (D) KEGG pathway, each row represents a functional entry, and each column represents an lncRNA. GSEA is a mode used to determine whether a given gene set has significant differences among different groups. Genes in these sets fill some degree of correlation. Therefore, enrichment analysis of gene sets can result up for the shortcomings of lone gene in the analysis.
qRT-PCR validation of differentially expressed mRNAs, lncRNAs, and circRNAs
We selected four DEmRNAs (Wbscr27, Sfrp5, Adig, and Saa3), DEcircRNAs- chr7:67264864–67268400:- and DElncRNA-ENSMUST00000169194, which are most apposite to obesity, for consume in verifying RNA-seq results using qRT-PCR. Results showed that the expression levels of Wbscr27, Sfrp5, Adig, and chr7:67264864–67268400:- were up-regulated in the EP-S group compared with the EP-M group, consistent with the sequencing results. The expressions of ENSMUST00000169194 and Saa3 were down-regulated in the EP-S group, which was moreover consistent with the sequencing results (Fig. 3).
Figure 3: Sequencing and quantitative PCR.
Sequencing and quantitative PCR for mRNAs (Wbscr27, Sfrp5, Adig, and Saa3), circRNAs- chr7:67264864–67268400:- and lncRNA-ENSMUST00000169194. The quantitative PCR results were consistent with the sequencing data. n = 5.
LncRNA–mRNA regulatory analysis
LncRNA has the competence to regulate several groups of mRNAs at the transcriptional level, including positive and negative regulation. Hence, understanding how Sal B alters the expression of lncRNA and its target mRNA expression is key to understanding its molecular mechanism of anti-obesity. They screened two DElncRNAs (ENSMUST0000140351 and ENSMUST00000169194) to construct the lncRNA–mRNA regulatory network map. A total of 11 differentially expressed mRNAs were establish to breathe negatively correlated with the expression of ENSMUST0000140351 (Ms4a14, Cd300ld3, Lum, Glipr1, Cd300ld5, Rgs18, Cd300ld4, Fgd4, Hpgds, Frmd4b, and Osbpl8). They moreover establish that the expressions of S100a8 and Fgl2 were positively correlated with the expression of ENSMUST00000169194 (Fig. 4; Table S4).
Figure 4: LncRNA–mRNA regulatory network (ENSMUST0000140351 and ENSMUST00000169194).
Squares picture lncRNAs, circles picture mRNAs; red indicates up-regulated expression and blue indicates down-regulated expression. The solid line is positively correlated and the dotted line is negatively correlated. LncRNA–mRNA regulatory network was constructed using Cytoscape v2.8.2 software.
Functional analysis of DEmRNAs
We performed travel and KEGG enrichment analysis to determine the functional significance of DEmRNAs in the EP-S/EP-M group. travel enrichment analysis showed that up-regulated mRNA was enriched in 13 BP, 5 CC, and 2 MF, while down-regulated mRNA was enriched in 89 BP, 24 CC, and 35 MF (Fig. 5; Table S5). The most highly enriched up-regulated travel terms were “brown stout cell differentiation (biological process),” “integral component of membrane (CC),” and “ligase activity (MF),” while the most highly enriched down-regulated travel terms were “immune system process (BP),” “extracellular region (CC),” and “chemokine activity (MF).”
Figure 5: travel analysis.
(A) up-regulated and (B) down-regulated of DEmRNAs. Using travel database (http://www.geneontology.org) analysis the travel enrichment of the DEmRNAs, based on three aspects: biological processes (BP), cellular components (CC), and molecular functions (MF). The log 10 values (p-value) denote enrichment scores and picture the significance of the travel term enrichment among the DEmRNAs.
KEGG pathway analysis showed that differentially expressed mRNAs were enriched in 14 pathways associated with obesity (p < 0.05). The most highly enriched pathways were “insulin resistance” and “IL-17 signaling pathway.” Notably, they moreover identified mRNAs involved in the regulation of these pathways, which may provide targets for Sal B as a potential drug to obviate HFD-induced obesity (Table 3). Moreover, a number of metabolic-related pathways were screened, including the “NF-κB signaling pathway” and the “B-cell receptor signaling pathway.” KEGG pathway analysis suggests the possibility that Sal B exerts its weight reduction result through the regulation of adipose metabolism via insulin resistance and IL-17 signaling in HFD induced-obesity mice.
KEGG pathway analysis.
IL-17 signaling pathway
Ccl12, Ccl7, Cxcl1, Fosl1, S100a8, S100a9
Atp6v0d2, Comp, Cybb, Fcgr1, Fcgr4, Msr1, Rab7b
Atp6v0d2, Ccl12, Ccl3, Cd86, Il18
NF-kappa B signaling pathway
Bcl2a1a, Bcl2a1b, Bcl2a1d, Btk, Card11
Btk, Fcgr1, Fcgr4, Fosl1, Lilra5
B cell receptor signaling pathway
Btk, Card11, Cd72, Rasgrp3
Toll-like receptor signaling pathway
Ccl3, Cd86, Tlr1, Tlr8
Cytokine-cytokine receptor interaction
Ccl12, Ccl3, Ccl7, Cxcl1, Cxcl16, Il18
Chemokine signaling pathway
Ccl12, Ccl3, Ccl7, Cxcl1, Cxcl16
NOD-like receptor signaling pathway
Ccl12, Cxcl1, Cybb, Il18
TNF signaling pathway
Ccl12, Cxcl1, Gm5431
Bcl2a1a, Bcl2a1b, Bcl2a1d
AGE–RAGE signaling pathway
Epigenetic alteration refers to a heritable change in gene expression under conditions in which the genomic DNA sequence does not change, resulting in an altered phenotype. This includes changes in the expression of non-coding RNA (Takada, Kouzmenko & Kato, 2009). Studies fill shown that epigenetic modification plays an primary role in the happening and evolution of obesity (Kasinska, Drzewoski & Sliwinska, 2016; Huang et al., 2018). With the advancement of RNA sequencing technology, more and more non-coding RNAs related to energy metabolism are recognized as involved in obesity and related metabolic diseases. WAT is mainly distributed in the subcutaneous tissue, omentum and mesentery of mice, with epididymis white stout (EP) as the most commonly used WAT in adipose studies. They studied the effects of Sal B on the expression of mRNAs, lncRNAs, and circRNAs in EP of HFD induced obesity mice from an epigenetic level, and explored the anti-obesity result of Sal B.
Obesity is an inflammatory condition that occurs in adipose tissue, and therefore constitutes a chronic inflammatory disease accompanied by activation of inflammatory signaling pathways in adipose tissue cells, release of inflammatory cytokines, and infiltration of immune cells (Nteeba et al., 2013; Mathieu, Lemieux & Després, 2010). Therefore, research on the treatment of obesity inflammation and fresh target exploitation will provide novel targets for the treatment of obesity and its related metabolic diseases. In the present study, they establish that the expression of many inflammation-associated mRNAs was affected by Sal B treatment, including Sfrp5 and Saa3. Secreted frizzled-related protein-5 (Sfrp5), known as an anti-inflammatory adipokine, is much more abundant in adipose tissue than other tissues, and negatively affects obesity and obesity-related metabolic disorders (Hu et al., 2013). Previous studies fill shown that in the adipose tissue of Sfrp5 knockout mice, the number of macrophages is significantly increased, and the expression of factors related to cellular inflammatory activity, such as TNF-a and IL-6, are significantly increased (Ouchi et al., 2010). Sfrp5 exerts anti-inflammatory effects by binding to Wnt5a to inhibit the activation of the downstream target JNK of the Wnt pathway and reduce the secretion of inflammatory factors in obese mice (Catalán et al., 2014) as well as in 3T3-L1 cells (Shadid & Jensen, 2003). Consistent with previous research, they establish that Sfrp5 is up-regulated 1.9-fold under Sal B treatment. Therefore, they hypothesize that Sal B may exert anti-obesity effects by regulating the expression of inflammation-related mRNA in adipose tissue. In addition, Sfrp5 can breathe used as a candidate target for studying the anti-obesity mechanism of Sal B, and its specific mechanism should breathe the stress of future research.
Another inflammation-related mRNA, Saa3, was establish to breathe expressed at half the control rate under Sal B treatment, consistent with previous studies. The serum amyloid A family is a class of proteins released during acute inflammatory response and is closely related to the pathogenesis of chronic inflammatory diseases such as obesity (Van Dielen et al., 2001). The expression of Saa3 is significantly increased under a promoted adipocyte inflammatory response by saturated fatty acids and glucose. In addition, Saa3 is highly expressed in the adipose tissue of obese mice, which may breathe related to the induction of adipose tissue inflammation (Den Hartigh et al., 2014). In this study, they establish that the expression of Saa3 was significantly down-regulated in EP-S, suggesting that Sal B can reduce the inflammatory response induced by Saa3.
In the EP-M group, they establish some abnormal expression of mRNA associated with adipose transformation, with Sal B intervention reversing these changes. Adipoietin (Adig), moreover known as tiny adipocytokines 1, plays a significant role in the differentiation of adipocytes (Ren et al., 2016). Their results showed that the expression Adig in the Sal B treatment group was significantly higher than in the obese model group. This is in line with the previous findings that Adig can promote the differentiation of adipocytes by activating the expression of PPARγ, Srebp-1, and Fas genes (Mei, Zhang & Fu, 2016). Therefore, they speculate that Sal B can up-regulate the expression level of Adig to activate the adipose transcription factor and promote the differentiation of adipocytes.
We identified 234 DElncRNAs, including 87 up-regulations and 147 down-regulations. Several differentially expressed lncRNAs might participate in lipid metabolism and glucose metabolism. For example, Rora and Dnm2 were predicted to breathe involved in “oxidative phosphorylation” and “glycine, serine, and threonine metabolism.” This is consistent with previous studies showing that Rora regulates lipid metabolism (Kim et al., 2017). Therefore, the differential expression profiles obtained attest that Sal B can exert a potential regulatory office in stout deposition and metabolism in obese mice by modulating the expression of lncRNAs.
Gene Ontology and KEGG pathway analyses were performed to foretell the viable functions of DEmRNAs. Their results indicated that the up-regulated expression of mRNAs involved in BP is primarily associated with brown adipocyte differentiation (Adig and Slc2a4), lipid metabolic process (Aacs, Fdx1, and Slc27a1), and metabolic process (Aacs and Slc27a1). The down-regulated mRNAs might breathe related to inflammatory response (Ccl12, Ccl3, Ccl7, Cd180, Cxcl1, Cybb, Il18, Ly86, S100a8, S100a9, Tlr1, and Tlr8). KEGG pathway analysis showed that two DEmRNAs, Slc27a1 and Slc2a4, were involved in the insulin resistance signaling pathway, which is closely related to obesity (Yu, Kim & Lee, 2017). The inflammatory signaling, IL-17 signaling, and NF-kappa B signaling pathways were moreover subjected to KEGG annotation prediction analysis (Tanti et al., 2012, Tarantino et al., 2014). Their results attest that Sal B may exert anti-obesity effects by modulating the expression of mRNAs in lipid metabolism and inflammation-related signaling pathways.
Among totality RNAs, circRNAs are the least understood, but issue to fill tangled regulatory effects in the evolution of obesity. They identified 9 up-regulated and 10 down-regulated circRNAs under Sal B treatment. Among them, the expression changes of chr14:103252408–103276518:- and chr14:103282597–103291362:- were the most obvious, with an approximate 30-fold upregulation. Therefore, these circRNAs may serve as targets for the design of therapeutic drugs for obesity, pending study of their specific mechanisms.
This study is the first comprehensive analysis of mRNA, lncRNA, and circRNA expression in EP of HFD-induced obese mice. They establish that Sal B regulates the expression of mRNAs and lncRNAs associated with adipocyte differentiation, lipid metabolism, and inflammation, as well as the insulin resistance and IL-17 signaling pathways. These findings imply that Sal B plays an primary role in inhibiting obesity by regulating anti-inflammatory related factors and signaling pathways. Their research provides valuable insights into the molecular mechanism of Sal B in anti-obesity effects and contributes therapeutic markers for pharmaceutical design in the prevention and treatment of obesity. In the future, the corresponding roles and molecular mechanisms of non-coding RNAs should breathe further elucidated. In addition, differential expression at the RNA level does not necessarily attest that the expression of related proteins is moreover significantly different. They aim to correlate RNA and protein analysis to more fully investigate the anti-obesity mechanism of Sal B.
From March to September 2014, 52 consecutive children (age scope 1–26 months) visiting their tertiary pediatric allergy headquarters for recent happening (last 2–4 weeks) of signs or symptoms of suspected non-IgE-mediated CMA, or for ensue up visit after 6 months of exclusion diet upon a confirmed diagnosis of non-IgE-mediated CMA were evaluated and invited to participate in a cross sectional study. The exclusion criteria were: consume of pre- or probiotic products and/or antibiotics in the previous 4 weeks; history of cow’s milk-induced anaphylaxis and/or other IgE-mediated signs of food allergy; concomitant presence of other food allergies or allergic diseases, eosinophilic disorders of the gastrointestinal tract, chronic systemic diseases, congenital cardiac defects, lively tuberculosis, autoimmune diseases, immunodeficiency, chronic inflammatory bowel diseases, celiac disease, cystic fibrosis, metabolic diseases, lactose intolerance, malignancy, chronic pulmonary diseases or malformations of the gastrointestinal tract. Written informed consent was obtained from the parents/guardians of each subject. The diagnosis of non-IgE-mediated CMA was based on clinical history, negative result of skin prick test, and/or negative level of IgE serum-specific anti-cow’s milk proteins, and the results of a double blind placebo-controlled oral food challenge (DBPCFC)33,34. totality DBPCFC were performed in a double-blind, placebo-controlled manner in the outpatient clinic on 2 sever days with a 1-week interval. Parents of patients taking antihistamines were advised to withhold these medications for 72 hours before and during the challenge. Randomization and preparation of the challenges were performed by experienced dietitians who were not directly involved in the procedures. In detail, every 20 minutes, increasing doses (0.1, 0.3, 1, 3, 10, 30, and 100 mL) of fresh pasteurized cow’s milk containing 3.5% of stout or an amino acid formula were administered. full emergency equipment and medications (epinephrine, antihistamines, and steroids) were available. The results were assessed simultaneously by experienced pediatric allergists. Study subjects were scored for 9 items divided into 4 main categories on a 0 to 3-point scale (0, none; 1, light; 2, moderate; and 3, severe): (1) universal (decreased blood pressure plus tachycardia); (2) skin (rash and urticaria/angioedema); (3) gastrointestinal (nausea or repeated vomiting, crampy-like abdominal pain, and diarrhea); and (4) respiratory (sneezing or itching, nasal congestion or rhinorrhea, and stridor deriving from upper airway obstruction or wheezing). If at least 2 of the 3 physicians independently scored one detail at level 3 or 2 (or more) items at level 2, the test result was considered positive. Children were observed for up to 4 hours after the final dose and then discharged. In case of a positive DBPCFC result at any testing dose, the patient remained under observation until symptom resolution. If the patient did not prove any symptoms within the first 24 hours, parents were advised to provide a lone feed of 100 mL of the tested formula (verum or placebo) every day at home for 7 days. If any symptoms occurred during this period, the patients returned to the outpatient clinic on the same day. After 7 days of verum or placebo administration, the patients were examined, and the parents were interviewed at the center. Parents were asked to contact the headquarters if any symptoms occurred in the 7 days after the DBPCFC procedures to rule out false-negative challenge results. The challenge result was considered negative if the patient tolerated the entire challenge, including the observation period. Fifty-two CMA patients were evaluated. Four patients were excluded because of the presence of exclusion criteria, and 2 were excluded for the lack of informed consent. Therefore, 46 CMA patients were included in this study. According to disease condition and dietary treatment, CMA patients were divided in three groups: group 1 included patients with non-IgE-mediated CMA at diagnosis, before any therapeutic intervention and receiving measure formula (n = 23); group 2 (n = 9) included patients with diagnosis of non-IgE-mediated CMA after treatment for 6 months with an extensively hydrolyzed casein formula (EHCF; Nutramigen, Mead Johnson Nutrition, Evansville IN, US); group 3 (n = 14) included patients with diagnosis of non-IgE-mediated CMA after treatment for 6 months with EHCF added with the probiotic L. rhamnosus GG (EHCF + LGG; Nutramigen LGG, Mead Johnson Nutrition, Evansville IN, US). The specific formula consume was prescribed and adherence was checked according to the measure procedure adopted at their Center. Briefly, the parents received written instructions regarding the commercial name of the product and the formula preparation procedure. Then, the adherence to the treatment was checked monthly during the first 3 months of treatment and then every 6 months. Formula consume was evaluated at each time visit by dietitians, counseling parents about issues that could arise during the elimination diet and on how to reach the daily recommended intake for Italian children. This allowed the study staff to evaluate compliance with the formula and to ensure that the patients received an usurp quantity of formula to meet their nutritional requirements. During the same study period, consecutive well children (group 4, n = 23), with negative clinical history for any allergic condition visiting their headquarters because of minimal surgical procedures or vaccination program were moreover enrolled. Anamnestic, demographic, anthropometric and clinical data were obtained from the parents of each theme and recorded in a clinical database. The 3-day dietary diary was collected from totality study subjects at enrolment. totality diaries were assessed using a specific software (Winfood, Medimatica srl, Colonnella, Teramo, Italy). For totality study subjects, a stool sample (3 g) was collected to evaluate gut microbiota composition and fecal butyrate concentration and stored at −80 °C until analyses.
The study was approved by the Ethics Committee of the University of Naples Federico II and was registered in the Clinical Trials Protocol Registration System on March 14, 2014 (https://clinicaltrials.gov - ID number: NCT02087930).
All methods were performed in accordance with the apposite guidelines and regulations.
DNA extraction and 16S sequencing
Fecal samples (about 1 g) were fully homogenized in STE buffer (100 mMNaCl, 10 mMTris-Cl pH 8.0, 1 mM EDTA pH 8.0) and centrifuged (500 × g, 1 min) in order to pellet debris. The supernatant was centrifuged again (12,000 × g, 2 min) and the pellet was used for DNA extraction with the PowerFecal DNA Isolation kit (Mo Bio Laboratories, Inc., Carlsbad, CA). V3-V4 region of the 16S rRNA gene was amplified by using primer and PCR conditions recently described35. PCR products were purified with the Agencourt AMPure XP beads (Beckman Coulter) and quantified using a Plate Reader AF2200 (Eppendorf). Amplicon multiplexing, pooling and sequencing were carried out following the Illumina 16S Metagenomic Sequencing Library Preparation protocol, on a MiSeq platform and using the MiSeq Reagent kit v2, leading to 2 × 250 bp, paired-end reads.
Fecal butyrate analysis
One gram of frozen feces was diluted with saline buffer, vortexed and centrifuged (12,000 × g) for 10 min in 2 ml tubes. The supernatant was filtered (0.45 μm) and stored at −20 °C until analysis. Frozen fecal extracts were acidified with 20 μl of 85% (w/v) phosphoric acid and 0.5 ml of ethyl acetate, mixed, centrifuged (12,000 × g) for 1 h, and extracted in duplicate. About 0.5 ml of the pooled extract containing the acidified butyrate was transferred into a 2 ml glass vial and loaded onto an Agilent Technologies (Santa Clara, CA, USA) 7890 gas chromatograph (GC) system with automatic loader/injector. The GC column was an Agilent J&W DB-FFAP (Agilent Technologies) of 30 m, internal diameter 0.25 mm and film thickness 0.25 μm. The GC was programmed to achieve the following sprint parameters: initial temperature 90 °C, hold 0.5 min, ramp of 20 °C min−1 up to a final temperature of 190 °C, total sprint time 8.0 min, gas flux 7.7 ml min−1 split less to maintain 3.26 p.s.i. column head pressure, septum purge 2.0 ml min−1. Detection was achieved using a flame ionization detector. Peaks were identified using a mixed external measure and quantified by peak height/internal measure ratio.
Statistical and bioinformatics analysis
All data were collected in a dedicated database and analysed by a statisticianwith IBM SPSS Statistics version 19.0 for Windows (SPSS Inc, Chicago, IL). The χ2 test and Fisher’s exact test were used for categorical variables. The level of significance for totality statistical tests was 2-sided, P < 0.05.
Raw sequence trait filtering and pre-processing was carried out as recently reported35. Briefly, demultiplexed, forward and invert reads were joined by using FLASH36. Joined reads were trait trimmed (Phred score < 20) and short reads (<250 bp) were discarded by using Prinseq37. towering trait reads were then imported in QIIME38. OTUs were picked through de novo approach and uclust mode and taxonomic assignment was obtained by using the RDP classifier and the Greengenes database39, following a pipeline previously reported35. In order to avoid biases deriving from different sequencing depth, OTU tables were rarefied to the lowest number of sequences per sample. Statistical analyses and visualization were carried out in R environment (https://www.r-project.org).
To discriminate the microbial profiles as a office of disease, a model based on projection on quiescent structures (PLS) in its discriminant (DA) version was built, based on the normalized abundance (log10) of the microbial genera identified. The R package mixOmics was used. Permutational Multivariate Analysis of Variance (non-parametric (PER)MANOVA) based on Jaccard and Bray Curtis distance matrices was applied with 999 permutations to detect significant differences in the overall microbial community composition, by using the adonis office in vegan package. Non-parametric Kruskal-Wallis and pairwise Wilcoxon tests were carried out in order to find OTUs differentially abundant between the groups. A Generalized Linear Model (R office glm) was built in order to test the consequence of continuous or discrete variables available for the subjects (mode of birth, age at weaning, age at sampling, sex, months of exclusive breastfeeding, tolerable daily consumption of proteins and fat, health status – that is, well or CMA) on the relative abundance of bacterial genera significantly different between well and CMA subjects. Spearman’s pairwise correlations were computed between OTUs or oligotypes and short-chain fatty acid abundance (corr.test office in psych package). Correction of p-values for multiple testing was performed40. Differences in fecal butyrate levels between the groups were evaluated by non-parametric Kruskal-Wallis and pairwise Wilcoxon tests. In order to compare the gut microbiota composition in children with non-IgE (analyzed in the present study) and IgE-mediated CMA from their previous study15, trait filtered reads of the previous study were downloaded from MG-RAST. Since the reads from the previous study included only V4 region of the 16S rRNAgene, they were aligned to those produced in this study, that were trimmed in 5′direction to the same length. Reads from both the studies were re-analysed as described above.
Sub-genus diversity of Bacteroides
Reads assigned to Bacteroides genus were extracted and entropy analysis and oligotyping41 were carried out as described previously42. After the initial round of oligotyping, towering entropy positions were chosen (−C option): 2, 30, 94, 104, 106, 107, 109, 114, 302, 380. To minimize the repercussion of sequencing errors, they required an oligotype to breathe represented by at least 100 reads (−M option). Moreover, rare oligotypes present in less than 10 samples were discarded (−s option). These parameters led 70,142 sequences left in the dataset. BLASTn was used to query the representative sequences against the NCBI nr database, and the top hit was considered for taxonomic assignment. Statistical analyses and visualization were carried out in R environment as described above.
The 16S rRNA gene sequences produced in this study are available at the Sequence Read Archive (SRA) of the National headquarters for Biotechnology Information (NCBI), under accession number SRP092171.