scholarly journals Temporary adhesion of the proseriate flatworm Minona ileanae

2019 ◽  
Vol 374 (1784) ◽  
pp. 20190194 ◽  
Author(s):  
Robert Pjeta ◽  
Julia Wunderer ◽  
Philip Bertemes ◽  
Teresa Hofer ◽  
Willi Salvenmoser ◽  
...  
Keyword(s):  
Cost Effective ◽  
Adhesive System ◽  
Ventral Side ◽  
Adhesive Properties ◽  
Expression Screening ◽  
Antibody Staining ◽  
A Genome ◽  
Tail Plate ◽  
Adhesive Organs ◽  
Adhesive Proteins

Flatworms can very rapidly attach to and detach from many substrates. In the presented work, we analysed the adhesive system of the marine proseriate flatworm Minona ileanae . We used light-, scanning- and transmission electron microscopy to analyse the morphology of the adhesive organs, which are located at the ventral side of the tail-plate. We performed transcriptome sequencing and differential RNA-seq for the identification of tail-specific transcripts. Using in situ hybridization expression screening, we identified nine transcripts that were expressed in the cells of the adhesive organs. Knock-down of five of these transcripts by RNA interference led to a reduction of the animal's attachment capacity. Adhesive proteins in footprints were confirmed using mass spectrometry and antibody staining. Additionally, lectin labelling of footprints revealed the presence of several sugar moieties. Furthermore, we determined a genome size of about 560 Mb for M. ileanae . We demonstrated the potential of Oxford Nanopore sequencing of genomic DNA as a cost-effective tool for identifying the number of repeats within an adhesive protein and for combining transcripts that were fragments of larger genes. A better understanding of the molecules involved in flatworm bioadhesion can pave the way towards developing innovative glues with reversible adhesive properties. This article is part of the theme issue ‘Transdisciplinary approaches to the study of adhesion and adhesives in biological systems’.

scholarly journals Proteins and microRNAs are differentially expressed in tear fluid from patients with Alzheimer’s disease

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Aidan Kenny ◽  
Eva M. Jiménez-Mateos ◽  
María Ascensión Zea-Sevilla ◽  
Alberto Rábano ◽  
Pablo Gili-Manzanaro ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Unmet Need ◽  
Cost Effective ◽  
Specific Protein ◽  
Tear Fluid ◽  
Initiation Factor ◽  
Non Invasive ◽  
A Genome ◽  
Potential Biomarker

Abstract Alzheimer’s disease (AD) is characterized by a progressive loss of neurons and cognitive functions. Therefore, early diagnosis of AD is critical. The development of practical and non-invasive diagnostic tests for AD remains, however, an unmet need. In the present proof-of-concept study we investigated tear fluid as a novel source of disease-specific protein and microRNA-based biomarkers for AD development using samples from patients with mild cognitive impairment (MCI) and AD. Tear protein content was evaluated via liquid chromatography-mass spectrometry and microRNA content was profiled using a genome-wide high-throughput PCR-based platform. These complementary approaches identified enrichment of specific proteins and microRNAs in tear fluid of AD patients. In particular, we identified elongation initiation factor 4E (eIF4E) as a unique protein present only in AD samples. Total microRNA abundance was found to be higher in tears from AD patients. Among individual microRNAs, microRNA-200b-5p was identified as a potential biomarker for AD with elevated levels present in AD tear fluid samples compared to controls. Our study suggests that tears may be a useful novel source of biomarkers for AD and that the identification and verification of biomarkers within tears may allow for the development of a non-invasive and cost-effective diagnostic test for AD.

An International Campaign for Agricultural and Livestock Genomics (CALG)

2002 ◽  
Vol 06 (24) ◽  
pp. 958-965
Author(s):  
Jun Yu ◽  
Jian Wang ◽  
Huanming Yang
Keyword(s):  
Large Scale ◽  
Cost Effective ◽  
Model Organisms ◽  
Environmental Biology ◽  
Cdna Sequences ◽  
Governmental Agencies ◽  
Technology Innovations ◽  
A Genome ◽  
Starting Point ◽  
The Cost

A coordinated international effort to sequence agricultural and livestock genomes has come to its time. While human genome and genomes of many model organisms (related to human health and basic biological interests) have been sequenced or plugged in the sequencing pipelines, agronomically important crop and livestock genomes have not been given high enough priority. Although we are facing many challenges in policy-making, grant funding, regional task emphasis, research community consensus and technology innovations, many initiatives are being announced and formulated based on the cost-effective and large-scale sequencing procedure, known as whole genome shotgun (WGS) sequencing that produces draft sequences covering a genome from 95 percent to 99 percent. Identified genes from such draft sequences, coupled with other resources, such as molecular markers, large-insert clones and cDNA sequences, provide ample information and tools to further our knowledge in agricultural and environmental biology in the genome era that just comes to its accelerated period. If the campaign succeeds, molecular biologists, geneticists and field biologists from all countries, rich or poor, would be brought to the same starting point and expect another astronomical increase of basic genomic information, ready to convert effectively into knowledge that will ultimately change our lives and environment into a greater and better future. We call upon national and international governmental agencies and organizations as well as research foundations to support this unprecedented movement.

Fixation and staining of Drosophila L1 larval brains for immunofluorescence microscopy and preparation for live imaging

2020 ◽  
Author(s):  
Phuong Thao Ly ◽  
John D. Thompson ◽  
Sharyn A. Endow
Keyword(s):  
Immunofluorescence Microscopy ◽  
Cost Effective ◽  
Live Imaging ◽  
Permeable Membrane ◽  
Antibody Staining ◽  
First Instar ◽  
Stem Cell Quiescence ◽  
Cell Quiescence ◽  
3D Printed ◽  
Entire Procedure

Abstract Drosophila first instar (L1) larval brains (LBs) contain frequent quiescent neural stem cells (qNSCs) as well as activated neuroblasts, making them favorable for studying stem cell quiescence and activation. However, the small size of LBs at the L1 stage necessitates the use of modified methods to prepare the LBs for immunofluorescence microscopy (IFM). The protocol described here allows efficient collection of embryos and maturation of larvae to the mid-L1 stage, followed by dissection, fixation and processing of LBs through the antibody staining steps for IFM. The entire procedure can be completed in ~3-5 days. Methods are also described for use in preparing L1 brains for live imaging experiments, including a file to create accessible and cost-effective 3D printed slides that can be fit with an O2-permeable membrane for live imaging.

scholarly journals Dentin Bonding and SEM Analysis of a New Experimental Universal Adhesive System Containing a Dendrimer

Polymers ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 461 ◽  
Author(s):  
Joana Vasconcelos e Cruz ◽  
Mário Polido ◽  
José Brito ◽  
Luisa L. Gonçalves
Keyword(s):  
Mixed Models ◽  
Dental Materials ◽  
Adhesive System ◽  
Sem Analysis ◽  
New Paradigm ◽  
Adhesive Properties ◽  
Universal Adhesive ◽  
Cross Sectional ◽  
Universal Adhesives ◽  
Self Etch

Due to their polymerization characteristics, hyper-branched dendrimers have lately shown to be promising candidates for use in dental materials. In this study, a new dental adhesive system was prepared, using a dendrimer derived from 2-isocyanatoethyl methacrylate (G-IEMA), and its adhesive properties were investigated. The exposed dentin was treated with four universal adhesives (UAs): SBU (Scotchbond Universal™), FUT (Futurabond M+™), AE1 (experimental adhesive with Bis-GMA) and AE2 (experimental adhesive with G-IEMA), using Etch & Rinse (ER) or Self Etch (SE) protocols. Composite build-ups were prepared and stored for 24 h at 37 °C in distilled water. Composite/dentin beams were prepared with cross-sectional areas of 1 ± 0.3 mm2 and µTBS (Micro-tensile bond strength) test was performed at 0.5 mm/min. Failures modes were evaluated by stereomicroscopy, and bonding interfaces were observed by scanning electron microscopy (SEM). Statistical analysis of µTBS data was performed using General Linear (GLM) and Linear Mixed Models (LMM). The effect of adhesive type on µTBS was significant (p = 0.010), with AE1 presenting significantly higher µTBS than SBU (p = 0.019). No other differences between adhesives were observed. ER showed significantly better results than SE (p = 0.019), and no significant interactions between the adhesives and protocols were determined. Results obtained so far pinpoint the emergence of a new paradigm in the dental materials field, as G-IEMA can be used successfully as an alternative to Bis-GMA.

scholarly journals Complete biosynthetic pathways of ascofuranone and ascochlorin inAcremonium egyptiacum

2019 ◽  
Vol 116 (17) ◽  
pp. 8269-8274 ◽  
Author(s):  
Yasuko Araki ◽  
Takayoshi Awakawa ◽  
Motomichi Matsuzaki ◽  
Rihe Cho ◽  
Yudai Matsuda ◽  
...  
Keyword(s):  
Differential Expression Analysis ◽  
Cost Effective ◽  
Drug Candidate ◽  
P450 Monooxygenase ◽  
Genome Wide ◽  
Common Precursor ◽  
A Genome ◽  
Membrane Bound ◽  
The Cost

Ascofuranone (AF) and ascochlorin (AC) are meroterpenoids produced by various filamentous fungi, includingAcremonium egyptiacum(synonym:Acremonium sclerotigenum), and exhibit diverse physiological activities. In particular, AF is a promising drug candidate against African trypanosomiasis and a potential anticancer lead compound. These compounds are supposedly biosynthesized through farnesylation of orsellinic acid, but the details have not been established. In this study, we present all of the reactions and responsible genes for AF and AC biosyntheses inA. egyptiacum, identified by heterologous expression, in vitro reconstruction, and gene deletion experiments with the aid of a genome-wide differential expression analysis. Both pathways share the common precursor, ilicicolin A epoxide, which is processed by the membrane-bound terpene cyclase (TPC) AscF in AC biosynthesis. AF biosynthesis branches from the precursor by hydroxylation at C-16 by the P450 monooxygenase AscH, followed by cyclization by a membrane-bound TPC AscI. All genes required for AC biosynthesis (ascABCDEFG) and a transcriptional factor (ascR) form a functional gene cluster, whereas those involved in the late steps of AF biosynthesis (ascHIJ) are present in another distantly located cluster. AF is therefore a rare example of fungal secondary metabolites requiring multilocus biosynthetic clusters, which are likely to be controlled by the single regulator, AscR. Finally, we achieved the selective production of AF inA. egyptiacumby genetically blocking the AC biosynthetic pathway; further manipulation of the strain will lead to the cost-effective mass production required for the clinical use of AF.

scholarly journals Use of Targeted Amplicon Sequencing in Peanut to Generate Allele Information on Allotetraploid Sub-Genomes

Genes ◽  
2020 ◽  
Vol 11 (10) ◽  
pp. 1220
Author(s):  
Roshan Kulkarni ◽  
Ratan Chopra ◽  
Jennifer Chagoya ◽  
Charles E. Simpson ◽  
Michael R. Baring ◽  
...  
Keyword(s):  
Cost Effective ◽  
Amplicon Sequencing ◽  
Routine Practice ◽  
Specific Information ◽  
Illumina Hiseq ◽  
Breeding Programs ◽  
Polymorphic Snps ◽  
Cost Effective Approach ◽  
A Genome ◽  
The Cost

The use of molecular markers in plant breeding has become a routine practice, but the cost per accession can be a hindrance to the routine use of Quantitative Trait Loci (QTL) identification in breeding programs. In this study, we demonstrate the use of targeted re-sequencing as a proof of concept of a cost-effective approach to retrieve highly informative allele information, as well as develop a bioinformatics strategy to capture the genome-specific information of a polyploid species. SNPs were identified from alignment of raw transcriptome reads (2 × 50 bp) to a synthetic tetraploid genome using BWA followed by a GATK pipeline. Regions containing high polymorphic SNPs in both A genome and B genomes were selected as targets for the resequencing study. Targets were amplified using multiplex PCR followed by sequencing on an Illumina HiSeq. Eighty-one percent of the SNP calls in diploids and 68% of the SNP calls in tetraploids were confirmed. These results were also confirmed by KASP validation. Based on this study, we find that targeted resequencing technologies have potential for obtaining maximum allele information in allopolyploids at reduced cost.

scholarly journals Comparative Transcriptome Analysis of Iron and Zinc Deficiency in Maize (Zea mays L.)

Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1812
Author(s):  
Mallana Gowdra Mallikarjuna ◽  
Nepolean Thirunavukkarasu ◽  
Rinku Sharma ◽  
Kaliyugam Shiriga ◽  
Firoz Hossain ◽  
...  
Keyword(s):  
Differential Expression ◽  
Cost Effective ◽  
Regulatory Elements ◽  
Sub Saharan Africa ◽  
Zn Deficiency ◽  
Metal Transporters ◽  
Maize Cultivars ◽  
A Genome ◽  
Iron And Zinc ◽  
Sub Saharan

Globally, one-third of the population is affected by iron (Fe) and zinc (Zn) deficiency, which is severe in developing and underdeveloped countries where cereal-based diets predominate. The genetic biofortification approach is the most sustainable and one of the cost-effective ways to address Fe and Zn malnutrition. Maize is a major source of nutrition in sub-Saharan Africa, South Asia and Latin America. Understanding systems’ biology and the identification of genes involved in Fe and Zn homeostasis facilitate the development of Fe- and Zn-enriched maize. We conducted a genome-wide transcriptome assay in maize inbred SKV616, under –Zn, –Fe and –Fe–Zn stresses. The results revealed the differential expression of several genes related to the mugineic acid pathway, metal transporters, photosynthesis, phytohormone and carbohydrate metabolism. We report here Fe and Zn deficiency-mediated changes in the transcriptome, root length, stomatal conductance, transpiration rate and reduced rate of photosynthesis. Furthermore, the presence of multiple regulatory elements and/or the co-factor nature of Fe and Zn in enzymes indicate their association with the differential expression and opposite regulation of several key gene(s). The differentially expressed candidate genes in the present investigation would help in breeding for Fe and Zn efficient and kernel Fe- and Zn-rich maize cultivars through gene editing, transgenics and molecular breeding.

scholarly journals Expression of Functional Recombinant Mussel Adhesive Protein Mgfp-5 in Escherichia coli

2004 ◽  
Vol 70 (6) ◽  
pp. 3352-3359 ◽  
Author(s):  
Dong Soo Hwang ◽  
Hyo Jin Yoo ◽  
Jong Hyub Jun ◽  
Won Kyu Moon ◽  
Hyung Joon Cha
Keyword(s):  
Escherichia Coli ◽  
Adhesion Force ◽  
Soluble Form ◽  
Material Surface ◽  
Adhesive Properties ◽  
Affinity Ligand ◽  
Mussel Adhesive Proteins ◽  
Mussel Adhesive ◽  
Aqueous Conditions ◽  
Adhesive Proteins

ABSTRACT Mussel adhesive proteins have been suggested as a basis for environmentally friendly adhesives for use in aqueous conditions and in medicine. However, attempts to produce functional and economical recombinant mussel adhesive proteins (mainly foot protein type 1) in several systems have failed. Here, the cDNA coding for Mytilus galloprovincialis foot protein type 5 (Mgfp-5) was isolated for the first time. Using this cDNA, we produced a recombinant Mgfp-5 fused with a hexahistidine affinity ligand, which was expressed in a soluble form in Escherichia coli and was highly purified using affinity chromatography. The adhesive properties of purified recombinant Mgfp-5 were compared with the commercial extracted mussel adhesive Cell-Tak by investigating adhesion force using atomic force microscopy, material surface coating, and quartz crystal microbalance. Even though further macroscale assays are needed, these microscale assays showed that recombinant Mgfp-5 has significant adhesive ability and may be useful as a bioadhesive in medical or underwater environments.

scholarly journals Cell attachment to thrombospondin: the role of ARG-GLY-ASP, calcium, and integrin receptors.

1988 ◽  
Vol 107 (6) ◽  
pp. 2351-2361 ◽  
Author(s):  
J Lawler ◽  
R Weinstein ◽  
R O Hynes
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Cell Attachment ◽  
Solid Phase ◽  
Rat Kidney ◽  
Muscle Cells ◽  
Cell Types ◽  
Adhesive Properties ◽  
Adhesive Proteins

Thrombospondin is a 420,000-D glycoprotein that has recently been shown to have several properties in common with the members of a class of adhesive proteins. To characterize further the adhesive properties of thrombospondin, we have studied its ability to support cell attachment. Thrombospondin adsorbed to plastic dishes supports the attachment of human endothelial and smooth muscle cells and the monocyte-like cell line (U937) as well as normal rat kidney cells. The majority of attached cells do not spread on the solid-phase thrombospondin. The attachment of all four cell types to thrombospondin is abolished if the assay is performed in the presence of EGTA, although the cells still attach to fibronectin. If thrombospondin is adsorbed to the dishes in the presence of EGTA and then washed with buffer containing calcium before addition of the cells, attachment is still markedly inhibited, indicating that calcium affects the conformation and function of thrombospondin. Attachment of all four cell types is also markedly inhibited by the synthetic peptides gly-arg-gly-asp-ser-pro (GRG-DSP) and gly-arg-gly-asp-ala-cys (GRGDAC) but not by the control peptide gly-arg-gly-glu-ser-pro (GRG-ESP). Affinity chromatography of n-octylglucoside extracts of surface-labeled endothelial cells or smooth muscle cells on thrombospondin-Sepharose and GRG-DSP-Affigel columns was used to identify an integrin complex related to glycoprotein IIb-IIIa as an RGD-dependent receptor for thrombospondin. In addition, a monoclonal antibody (LM609) that blocks attachment of endothelial cells to vitronectin, fibrinogen, and von Willebrand factor also inhibits attachment of endothelial cells to thrombospondin. These data indicate that the attachment of cells to thrombospondin is mediated by RGD and calcium-dependent mechanisms and is consistent with the hypothesis that the GRGDAC sequence in thrombospondin is a site for interaction with an integrin receptor of the beta 3 subclass.

scholarly journals (Un)expected Similarity of the Temporary Adhesive Systems of Marine, Brackish, and Freshwater Flatworms

2021 ◽  
Vol 22 (22) ◽  
pp. 12228
Author(s):  
Philip Bertemes ◽  
Robert Pjeta ◽  
Julia Wunderer ◽  
Alexandra L. Grosbusch ◽  
Birgit Lengerer ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridisation ◽  
Adhesive System ◽  
Wound Dressings ◽  
Freshwater Species ◽  
Freshwater Habitats ◽  
Anchor Cell ◽  
Adhesive Proteins ◽  
Related Proteins

Many free-living flatworms have evolved a temporary adhesion system, which allows them to quickly attach to and release from diverse substrates. In the marine Macrostomum lignano, the morphology of the adhesive system and the adhesion-related proteins have been characterised. However, little is known about how temporary adhesion is performed in other aquatic environments. Here, we performed a 3D reconstruction of the M. lignano adhesive organ and compared it to the morphology of five selected Macrostomum, representing two marine, one brackish, and two freshwater species. We compared the protein domains of the two adhesive proteins, as well as an anchor cell-specific intermediate filament. We analysed the gene expression of these proteins by in situ hybridisation and performed functional knockdowns with RNA interference. Remarkably, there are almost no differences in terms of morphology, protein regions, and gene expression based on marine, brackish, and freshwater habitats. This implies that glue components produced by macrostomids are conserved among species, and this set of two-component glue functions from low to high salinity. These findings could contribute to the development of novel reversible biomimetic glues that work in all wet environments and could have applications in drug delivery systems, tissue adhesives, or wound dressings.

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