Faecalibacterium prausnitzii Treatment Improves Hepatic Health and Reduces Adipose Tissue Inflammation in High-fat Fed Mice

Faecalibacterium prausnitzii is considered as one of the most important bacterial indicators of a healthy gut. We studied the effects of oral F. prausnitzii treatment on high-fat fed mice. Compared to the high-fat control mice, F. prausnitzii-treated mice had lower hepatic fat content, aspartate aminotransferase and alanine aminotransferase, and increased fatty acid oxidation and adiponectin signaling in liver. Hepatic lipidomic analyses revealed decreases in several species of triacylglycerols, phospholipids and cholesteryl esters. Adiponectin expression was increased in the visceral adipose tissue, and the subcutaneous and visceral adipose tissues were more insulin sensitive and less inflamed in F. prausnitzii-treated mice. Further, F. prausnitzii treatment increased muscle mass that may be linked to enhanced mitochondrial respiration, modified gut microbiota composition and improved intestinal integrity. Our findings show that F. prausnitzii treatment improves hepatic health, and decreases adipose tissue inflammation in mice and warrant the need for further studies to discover its therapeutic potential.</p

Novel small molecule in differentiation of hiPSC derived cardiomyocytes and single cell beating analysis

Sydäntaudit ovat johtava kuolinsyy maailmanlaajuisesti, ja siksi potilasspesifisien lääketestausmallien sekä kudosregeneraation tutkimus on ajankohtainen aihe. Nykyinen teknologia ei pysty vielä tuottamaan suuria määriä puhtaita ihmisen indusoiduista pluripotenteista kantasoluista (hiPSC) erilaistettuja sydänsoluja tarpeeksi tehokkaasti, jotta niitä voitaisiin käyttää regeneratiivisessa lääketieteessä ja luotettavammassa lääketestauksessa. Sydänsolujen tuottamista ja karakterisointia täytyy kehittää. Tämän tutkimuksen tavoite oli jatkaa potilasspesifisten hiPSC sydänsolujen erilaistamisen karakterisoimista ja testata oletetun sydänsolujen erilaistusta tehostavan pienmolekyylin (PCDE) vaikutusta erilaistamisen tehokkuuteen. Hypoteesi oli, että kyseinen pienmolekyyli parantaisi erilaistustehokkuutta. Erilaistustehokkuutta mitattiin virtaussytometrisesti sydänmarkkerilla, Troponiin T:llä, ja kardiomyosiitit karakterisoitiin immunovärjäyksillä sekä kvantitatiivisen polymeraasiketjureaktion (qPCR) avulla. Geeniekspressiota analysoitiin RNA-näytteistä erilaistuksen eri vaiheista. Tutkimuksen aikana arvioitiin tekniikkana videopohjaista yksittäisten solujen sykeanalyysia, joka on tietokoneohjelmaan pohjautuva tukeva tutkimusmenetelmä yksittäisten solujen sykekuvion määrittämiseksi. Tuloksiin perustuen PCDE ei onnistunut tehostamaan hiPSC sydänsolujen erilaistamista ja saattoi vähentää sydänmarkkereiden ekspressiota. Totesimme kuitenkin, että videopohjaista sykeanalyysiä voitaisiin käyttää yhdessä yksittäisten solujen PCR-tutkimusten kanssa sydänsolujen paremman karakterisoinnin saavuttamiseksi.Cardiac disease remains the leading cause of death in the world. Therefore, the need for patient specific drug testing models, as well as tissue regeneration, are hot topics in medicine. The modern technology is still unable to efficiently produce pure cultures of human induced stem cell (hiPSC) derived cardiomyocytes, which are needed for several purposes including regenerative medicine and reliable drug testing for cardiac disease. Further development of hiPSC derived cardiomyocyte differentiation and characterization techniques is required. The objective of this experiment was to further characterize the differentiation of hiPSC derived cardiomyocytes obtained from patients and to test the effects of a novel putative cardiac differentiation enhancing small molecule (PCDE), on the efficiency of differentiation. The hypothesis was that the small molecule would increase the differentiation efficiency. The efficiency was tested with flow cytometry using a cardiac marker, Troponin T, and the cardiomyocytes were characterized by immunostaining and quantitative polymerase chain reaction (qPCR). Gene expression was analyzed from RNA samples collected from different time points of differentiation. During the experiment video-based beating analysis, a robust technique that utilizes a computer program to characterize beating patterns of single cardiomyocytes, was evaluated as a technique. Based on all results PCDE failed to enhance the differentiation efficiency of hiPSC derived cardiomyocytes and may have reduced the expression of cardiac markers. However, we found that the video based beating analysis could be used in combination with single cell PCR to better characterize cardiomyocytes

High throughput generation of a resource of the human secretome in mammalian cells

The proteins secreted by human tissues and blood cells, the secretome, are important both for the basic understanding of human biology and for identification of potential targets for future diagnosis and therapy. Here, a high-throughput mammalian cell factory is presented that was established to create a resource of recombinant full-length proteins covering the majority of those annotated as ‘secreted’ in humans. The full-length DNA sequences of each of the predicted secreted proteins were generated by gene synthesis, the constructs were transfected into Chinese hamster ovary (CHO) cells and the recombinant proteins were produced, purified and analyzed. Almost 1,300 proteins were successfully generated and proteins predicted to be secreted into the blood were produced with a success rate of 65%, while the success rates for the other categories of secreted proteins were somewhat lower giving an overall one-pass success rate of ca. 58%. The proteins were used to generate targeted proteomics assays and several of the proteins were shown to be active in a phenotypic assay involving pancreatic β-cell dedifferentiation. Many of the proteins that failed during production in CHO cells could be rescued in human embryonic kidney (HEK 293) cells suggesting that a cell factory of human origin can be an attractive alternative for production in mammalian cells. In conclusion, a high-throughput protein production and purification system has been successfully established to create a unique resource of the human secretome.ISSN:1871-6784ISSN:1876-434

Contribution of Antibody-based Protein Profiling to the Human Chromosome-centric Proteome Project (C-HPP)

A gene-centric Human Proteome Project has been proposed to characterize the human protein-coding genes in a chromosome-centered manner to understand human biology and disease. Here, we report on the protein evidence for all genes predicted from the genome sequence based on manual annotation from literature (UniProt), antibody-based profiling in cells, tissues and organs and analysis of the transcript profiles using next generation sequencing in human cell lines of different origins. We estimate that there is good evidence for protein existence for 69% (n = 13985) of the human protein-coding genes, while 23% have only evidence on the RNA level and 7% still lack experimental evidence. Analysis of the expression patterns shows few regards to protein evidence is visualized in a chromosome-centric manner as part of a new version of the Human Protein Atlas (www.proteinatlas.org)

Tissue-based map of the human proteome

Protein expression across human tissues Sequencing the human genome gave new insights into human biology and disease. However, the ultimate goal is to understand the dynamic expression of each of the approximately 20,000 protein-coding genes and the function of each protein. Uhlén et al. now present a map of protein expression across 32 human tissues. They not only measured expression at an RNA level, but also used antibody profiling to precisely localize the corresponding proteins. An interactive website allows exploration of expression patterns across the human body. Science , this issue 10.1126/science.1260419 </jats:p

Faecalibacterium prausnitzii treatment improves hepatic health and reduces adipose tissue inflammation in high-fat fed mice

Faecalibacterium prausnitzii is considered as one of the most important bacterial indicators of a healthy gut. We studied the effects of oral F. prausnitzii treatment on high-fat fed mice. Compared to the high-fat control mice, F. prausnitzii-treated mice had lower hepatic fat content, aspartate aminotransferase and alanine aminotransferase, and increased fatty acid oxidation and adiponectin signaling in liver. Hepatic lipidomic analyses revealed decreases in several species of triacylglycerols, phospholipids and cholesteryl esters. Adiponectin expression was increased in the visceral adipose tissue, and the subcutaneous and visceral adipose tissues were more insulin sensitive and less inflamed in F. prausnitzii-treated mice. Further, F. prausnitzii treatment increased muscle mass that may be linked to enhanced mitochondrial respiration, modified gut microbiota composition and improved intestinal integrity. Our findings show that F. prausnitzii treatment improves hepatic health, and decreases adipose tissue inflammation in mice and warrant the need for further studies to discover its therapeutic potential.peerReviewe

A roadmap to generate renewable protein binders to the human proteome

Despite the wealth of commercially available antibodies to human proteins, research is often hindered by their inconsistent validation, their poor performance and the inadequate coverage of the proteome. These issues could be addressed by systematic, genome-wide efforts to generate and validate renewable protein binders. We report a multicenter study to assess the potential of hybridoma and phage-display technologies in a coordinated large-scale antibody generation and validation effort. We produced over 1,000 antibodies targeting 20 SH2 domain proteins and evaluated them for potency and specificity by enzyme-linked immunosorbent assay (ELISA), protein microarray and surface plasmon resonance (SPR). We also tested selected antibodies in immunoprecipitation, immunoblotting and immunofluorescence assays. Our results show that high-affinity, high-specificity renewable antibodies generated by different technologies can be produced quickly and efficiently. We believe that this work serves as a foundation and template for future larger-scale studies to create renewable protein binders