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Micro-organismen als regisseurs van de duurzame eiwittransitie

In een tijd waarin de wereld geconfronteerd wordt met een toenemende bevolking en de daaruit voortvloeiende behoefte aan voedsel, staat het lectoraat Eiwittransitie voor een uiterst relevante uitdaging. De groeiende vraag naar eiwitten en de noodzaak om onze consumptiegewoonten in balans te krijgen met natuur en onze gezondheid vormen de kern van de missie van dit lectoraat.

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Micro-organismen als regisseurs van de duurzame eiwittransitie

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PCR-DGGE method to assess the diversity of BTEX mono-oxygenase genes at contaminated sites

tmoA and related genes encode the alpha-subunit of the hydroxylase component of the major group (subgroup 1 of subfamily 2) of bacterial multicomponent mono-oxygenase enzyme complexes involved in aerobic benzene, toluene, ethylbenzene and xylene (BTEX) degradation. A PCR-denaturing gradient gel electrophoresis (DGGE) method was developed to assess the diversity of tmoA-like gene sequences in environmental samples using a newly designed moderately degenerate primer set suitable for that purpose. In 35 BTEX-degrading bacterial strains isolated from a hydrocarbon polluted aquifer, tmoA-like genes were only detected in two o-xylene degraders and were identical to the touA gene of Pseudomonas stutzeri OX1. The diversity of tmoA-like genes was examined in DNA extracts from contaminated and non-contaminated subsurface samples at a site containing a BTEX-contaminated groundwater plume. Differences in DGGE patterns were observed between strongly contaminated, less contaminated and non-contaminated samples and between different depths, suggesting that the diversity of tmoA-like genes was determined by environmental conditions including the contamination level. Phylogenetic analysis of the protein sequences deduced from the amplified amplicons showed that the diversity of TmoA-analogues in the environment is larger than suggested from described TmoA-analogues from cultured isolates, which was translated in the DGGE patterns. Although different positions on the DGGE gel can correspond to closely related TmoA-proteins, relationships could be noticed between the position of tmoA-like amplicons in the DGGE profile and the phylogenetic position of the deduced protein sequence.

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Site directed mutagenesis in Petunia using CRISPR/CAS9 technology

The Green Biotechnology research group focusses on the application of molecular breeding/biotechnological tools and also on the development/analysis of new tools, for the breeding of enhanced vegetable crops and ornamental plants. The research group is positioned within Inholland University of Applied Sciences, Life Sciences & Chemistry and serves as a link between the breeding companies and our education of the skilled technicians of tomorrow. We are working on the development of a method for targeted mutagenesis of plant genomes using the bacterial CRISPR-Cas system. This method greatly enhances the effectiveness and speed by which new crops and plants can be developed

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Site directed mutagenesis in Petunia using CRISPR/CAS9 technology

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Analysis of the fluorescent properties of vaginal fluid upon ageing

Detection and identification of body fluids are crucial aspects of forensic investigations, aiding in crime scene reconstructions and providing important leads. Although many methods have been developed for these purposes, no method is currently in use in the forensic field that allows rapid, non-contact detection and identification of vaginal fluids directly at the crime scene. The development of such technique is mainly challenged by the complex chemistry of the constituents, which can differ between donors and exhibits changes based on woman’s menstrual cycle. The use of fluorescence spectroscopy has shown promise in this area for other biological fluids. Therefore, the aim of this study was to identify specific fluorescent signatures of vaginal fluid with fluorescence spectroscopy to allow on-site identification. Additionally, the fluorescent properties were monitored over time to gain insight in the temporal changes of the fluorescent spectra of vaginal fluid. The samples were excited at wavelengths ranging from 200 to 600 nm and the induced fluorescence emission was measured from 220 to 700 nm. Excitation and emission maps (EEMs) were constructed for eight donors at seven time points after donation. Four distinctive fluorescence peaks could be identified in the EEMs, indicating the presence of proteins, fluorescent oxidation products (FOX), and an unidentified component as the dominant contributors to the fluorescence. To further asses the fluorescence characteristics of vaginal fluid, the fluorescent signatures of protein and FOX were used to monitor protein and lipid oxidation reactions over time. The results of this study provide insights into the intrinsic fluorescent properties of vaginal fluid over time which could be used for the development of a detection and identification method for vaginal fluids. Furthermore, the observed changes in fluorescence signatures over time could be utilized to establish an accurate ageing model.

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Analysis of the fluorescent properties of vaginal fluid upon ageing

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Highlights from the 7th European meeting on molecular diagnostics

This report presents the highlights of the 7th European Meeting on Molecular Diagnostics held in Scheveningen, The Hague, The Netherlands, 12-14 October 2011. The areas covered included molecular diagnostics applications in medical microbiology, virology, pathology, hemato-oncology,clinical genetics and forensics. Novel real-time amplification approaches, novel diagnostic applications and new technologies, such as next-generation sequencing, PCR lectrospray-ionization TOF mass spectrometry and techniques based on the detection of proteins or other molecules, were discussed. Furthermore, diagnostic companies presented their future visions for molecular diagnostics in human healthcare.

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Highlights from the 7th European meeting on molecular diagnostics

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Proteins and amino acids are fundamental to optimal nutrition support in critically ill patients

In this review, we present the growing scientific evidence showing the importance of protein and amino acid provision in nutritional support and their impact on preservation of muscle mass and patient outcomes.

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Proteins and amino acids are fundamental to optimal nutrition support in critically ill patients

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Alternative primer sets for PCR detection of genotypes involved in bacterial aerobic BTEX degradation

Eight new primer sets were designed for PCR detection of (i) mono-oxygenase and dioxygenase gene sequences involved in initial attack of bacterial aerobic BTEX degradation and of (ii) catechol 2,3-dioxygenase gene sequences responsible for meta-cleavage of the aromatic ring. The new primer sets allowed detection of the corresponding genotypes in soil with a detection limit of 10(3)-10(4) or 10(5)-10(6) gene copies g(-1) soil, assuming one copy of the gene per cell. The primer sets were used in PCR to assess the distribution of the catabolic genes in BTEX degrading bacterial strains and DNA extracts isolated from soils sampled from different locations and depths (vadose, capillary fringe and saturated zone) within a BTEX contaminated site. In both soil DNA and the isolates, tmoA-, xylM- and xylE1-like genes were the most frequently recovered BTEX catabolic genes. xylM and xylE1 were only recovered from material from the contaminated samples while tmoA was detected in material from both the contaminated and non-contaminated samples. The isolates, mainly obtained from the contaminated locations, belonged to the Actinobacteria or Proteobacteria (mainly Pseudomonas). The ability to degrade benzene was the most common BTEX degradation phenotype among them and its distribution was largely congruent with the distribution of the tmoA-like genotype. The presence of tmoA and xylM genes in phylogenetically distant strains indicated the occurrence of horizontal transfer of BTEX catabolic genes in the aquifer. Overall, these results show spatial variation in the composition of the BTEX degradation genes and hence in the type of BTEX degradation activity and pathway, at the examined site. They indicate that bacteria carrying specific pathways and primarily carrying tmoA/xylM/xylE1 genotypes, are being selected upon BTEX contamination.

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Novel proteins: insects and fish

Flyer with information about the lectureship INVIS, HAS Hogeschool. Extend and integrate knowledge, experience, and education on healthy and safe insect and fish culture: Investigate risk factors and support the use of healthy and safe insects in aquaculture feed in cooperation with feed processors.

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Enterocytes, fibroblasts and myeloid cells synergize in anti-bacterial and anti-viral pathways with IL22 as the central cytokine

IL22 is an important cytokine involved in the intestinal defense mechanisms against microbiome. By using ileum-derived organoids, we show that the expression of anti-microbial peptides (AMPs) and anti-viral peptides (AVPs) can be induced by IL22. In addition, we identified a bacterial and a viral route, both leading to IL22 production by T cells, but via different pathways. Bacterial products, such as LPS, induce enterocyte-secreted SAA1, which triggers the secretion of IL6 in fibroblasts, and subsequently IL22 in T cells. This IL22 induction can then be enhanced by macrophage-derived TNFα in two ways: by enhancing the responsiveness of T cells to IL6 and by increasing the expression of IL6 by fibroblasts. Viral infections of intestinal cells induce IFNβ1 and subsequently IL7. IFNβ1 can induce the expression of IL6 in fibroblasts and the combined activity of IL6 and IL7 can then induce IL22 expression in T cells. We also show that IL22 reduces the expression of viral entry receptors (e.g. ACE2, TMPRSS2, DPP4, CD46 and TNFRSF14), increases the expression of anti-viral proteins (e.g. RSAD2, AOS, ISG20 and Mx1) and, consequently, reduces the viral infection of neighboring cells. Overall, our data indicates that IL22 contributes to the innate responses against both bacteria and viruses.

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Products 39

Micro-organismen als regisseurs van de duurzame eiwittransitie

PCR-DGGE method to assess the diversity of BTEX mono-oxygenase genes at contaminated sites

Site directed mutagenesis in Petunia using CRISPR/CAS9 technology

Analysis of the fluorescent properties of vaginal fluid upon ageing

Highlights from the 7th European meeting on molecular diagnostics

Proteins and amino acids are fundamental to optimal nutrition support in critically ill patients

Alternative primer sets for PCR detection of genotypes involved in bacterial aerobic BTEX degradation

Novel proteins: insects and fish

Enterocytes, fibroblasts and myeloid cells synergize in anti-bacterial and anti-viral pathways with IL22 as the central cytokine

Projects 1

Mycobacterial Coating with Trehalose-Associated Proteins.

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Products 39

Micro-organismen als regisseurs van de duurzame eiwittransitie

PCR-DGGE method to assess the diversity of BTEX mono-oxygenase genes at contaminated sites

Site directed mutagenesis in Petunia using CRISPR/CAS9 technology

Analysis of the fluorescent properties of vaginal fluid upon ageing

Highlights from the 7th European meeting on molecular diagnostics

Proteins and amino acids are fundamental to optimal nutrition support in critically ill patients

Alternative primer sets for PCR detection of genotypes involved in bacterial aerobic BTEX degradation

Novel proteins: insects and fish

Enterocytes, fibroblasts and myeloid cells synergize in anti-bacterial and anti-viral pathways with IL22 as the central cytokine

Projects 1

Mycobacterial Coating with Trehalose-Associated Proteins.