Bridging the gap between healthcare sectors: Facilitating the transition from NICU to the municipality and home for families with premature infants.

Purpose: The transition from hospital to home can be challenging for parents of prematurely born infants. The aim of this ethnographic study was to describe a multidisciplinary and cross-sectoral discharge conference for families with premature infants transitioning from a neonatal intensive care unit to municipal healthcare services.

Design And Methods: An ethnographically/anthropologically inspired qualitative design was adopted.

"They aren't waiting for an SLP, they think 'what can I as a parent do now?'" course leaders' perceptions of AAC interventions targeting parents.

Parental interventions can help parents use strategies to support their child's language and communication development. The ComAlong courses are parental interventions that focus on responsive communication, enhanced milieu teaching, and augmentative and alternative communication. This interview study aimed to investigate the course leaders' perceptions of the three ComAlong courses, ComAlong Habilitation, ComAlong Developmental Language Disorder, and ComAlong Toddler, and to evaluate their experiences of the implementation of the courses.

Impact of a Conformité Européenne (CE) Certification-Marked Medical Software Sensor on COVID-19 Pandemic Progression Prediction: Register-Based Study Using Machine Learning Methods.

Background: To address the current COVID-19 and any future pandemic, we need robust, real-time, and population-scale collection and analysis of data. Rapid and comprehensive knowledge on the trends in reported symptoms in populations provides an earlier window into the progression of viral spread, and helps to predict the needs and timing of professional health care.

Objective: The objective of this study was to use a Conformité Européenne (CE)-marked medical online symptom checker service, Omaolo, and validate the data against the national demand for COVID-19-related care to predict the pandemic progression in Finland.

Nurses' experiences of screening for communication difficulties at 18 months of age.

Aim: Early identification of communication disorders is important and may be possible through screening in the child health services. The aim of the study was to investigate nurses' experiences and sense of competence when using the Infant-Toddler Checklist (ITC) communication screening at the 18-month health visit.

Methods: A mixed-methods design including three focus group interviews (n = 14) and a web-based survey (n = 22) among nurses using the ITC or the standard method.

Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein.

Background: Engineered proteins, with non-immunoglobulin scaffolds, have become an important alternative to antibodies in many biotechnical and therapeutic applications. When compared to antibodies, tailored proteins may provide advantageous properties such as a smaller size or a more stable structure.

Results: Avidin is a widely used protein in biomedicine and biotechnology.

Chimeric avidin shows stability against harsh chemical conditions--biochemical analysis and 3D structure.

Avidin and its bacterial analog streptavidin have been widely used in applications in life sciences. Recently, we described a highly thermostable engineered avidin, called chimeric avidin, which is a hybrid of avidin and avidin-related protein 4. Here, we report a protocol for pilot-scale production in E.

Structural and functional characteristics of xenavidin, the first frog avidin from Xenopus tropicalis.

Background: Avidins are proteins with extraordinarily high ligand-binding affinity, a property which is used in a wide array of life science applications. Even though useful for biotechnology and nanotechnology, the biological function of avidins is not fully understood. Here we structurally and functionally characterise a novel avidin named xenavidin, which is to our knowledge the first reported avidin from a frog.

Crystal structure of rhizavidin: insights into the enigmatic high-affinity interaction of an innate biotin-binding protein dimer.

Rhizavidin, from the proteobacterium Rhizobium etli, exhibits high affinity towards biotin but maintains an inherent dimeric quaternary structure and thus, differs from all other known tetrameric avidins. Rhizavidin also differs from the other avidins, since it lacks the characteristic tryptophan residue positioned in the L7,8 loop that plays a crucial role in high-affinity binding and oligomeric stability of the tetrameric avidins. The question is, therefore, how does the dimer exist and how is the high biotin-binding affinity retained? For this purpose, the crystal structures of apo- and biotin-complexed rhizavidin were determined.

Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes.

Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form).

Bradavidin II from Bradyrhizobium japonicum: a new avidin-like biotin-binding protein.

A gene encoding an avidin-like protein was discovered in the genome of B. japonicum. The gene was cloned to an expression vector and a protein, named bradavidin II, was produced in E.

Rational modification of ligand-binding preference of avidin by circular permutation and mutagenesis.

Chicken avidin is a key component used in a wide variety of biotechnological applications. Here we present a circularly permuted avidin (cpAvd4-->3) that lacks the loop between beta-strands 3 and 4. Importantly, the deletion of the loop has a positive effect on the binding of 4'-hydroxyazobenzene-2-carboxylic acid (HABA) to avidin.

Critical importance of loop conformation to avidin-enhanced hydrolysis of an active biotin ester.

The homotetrameric and biotin-binding properties of avidin and streptavidin have been exploited for a myriad of biotechnological applications and theoretical studies. Among the few differences between the two proteins is the capacity of avidin to hydrolyze biotinyl p-nitrophenyl ester (BNP), as opposed to streptavidin, which fully protects the same pseudosubstrate from hydrolysis. Combined mutagenesis and X-ray analysis have been used to attempt to understand this diametric difference in activities.

Carbonic anhydrase inhibitors: the very weak inhibitors dithiothreitol, beta-mercaptoethanol, tris(carboxyethyl)phosphine and threitol interfere with the binding of sulfonamides to isozymes II and IX.

The inhibition of the metalloenzyme carbonic anhydrase (CA, EC 4.2.1.

Rhizavidin from Rhizobium etli: the first natural dimer in the avidin protein family.

Rhizobium etli CFN42 is a symbiotic nitrogen-fixing bacterium of the common bean Phaseolus vulgaris. The symbiotic plasmid p42d of R. etli comprises a gene encoding a putative (strept)avidin-like protein, named rhizavidin.

Brave new (strept)avidins in biotechnology.

Avidin and streptavidin are widely used in (strept)avidin-biotin technology, which is based on their tight biotin-binding capability. These techniques are exceptionally diverse, ranging from simple purification and labeling methods to sophisticated drug pre-targeting and nanostructure-building approaches. Improvements in protein engineering have provided new possibilities to develop tailored protein tools.

Structure and characterization of a novel chicken biotin-binding protein A (BBP-A).

Background: The chicken genome contains a BBP-A gene showing similar characteristics to avidin family genes. In a previous study we reported that the BBP-A gene may encode a biotin-binding protein due to the high sequence similarity with chicken avidin, especially at regions encoding residues known to be located at the ligand-binding site of avidin.

Results: Here, we expand the repertoire of known macromolecular biotin binders by reporting a novel biotin-binding protein A (BBP-A) from chicken.

Genetically engineered avidins and streptavidins.

Chicken avidin and bacterial streptavidin, (strept)avidin, are proteins widely utilized in a number of applications in life science, ranging from purification and labeling techniques to diagnostics, and from targeted drug delivery to nanotechnology. (Strept)avidin-biotin technology relies on the extremely tight and specific affinity between (strept)avidin and biotin (dissociation constant, K(d) approximately 10(-14)-10(-16) M). (Strept)avidins are also exceptionally stable proteins.

Controlling quaternary structure assembly: subunit interface engineering and crystal structure of dual chain avidin.

Dual chain avidin (dcAvd) is an engineered avidin form, in which two circularly permuted chicken avidin monomers are fused into one polypeptide chain. DcAvd can theoretically form two different pseudotetrameric quaternary assemblies because of symmetry at the monomer-monomer interfaces. Here, our aim was to control the assembly of the quaternary structure of dcAvd.

Dual-affinity avidin molecules.

A recently reported dual-chain avidin was modified further to contain two distinct, independent types of ligand-binding sites within a single polypeptide chain. Chicken avidin is normally a tetrameric glycoprotein that binds water-soluble d-biotin with extreme affinity (K(d) approximately 10(-15) M). Avidin is utilized in various applications and techniques in the life sciences and in the nanosciences.

Production of Hev b5 as a fluorescent biotin-binding tripartite fusion protein in insect cells.

The presented green fluorescent protein and streptavidin core-based tripartite fusion system provides a simple and efficient way for the production of proteins fused to it in insect cells. This fusion protein forms a unique tag, which serves as a multipurpose device enabling easy optimization of production, one-step purification via streptavidin-biotin interaction, and visualization of the fusion protein during downstream processing and in applications. In the present study, we demonstrate the successful production, purification, and detection of a natural rubber latex allergen Hev b5 with this system.

Tetravalent single-chain avidin: from subunits to protein domains via circularly permuted avidins.

scAvd (single-chain avidin, where two dcAvd are joined in a single polypeptide chain), having four biotin-binding domains, was constructed by fusion of topologically modified avidin units. scAvd showed similar biotin binding and thermal stability properties as chicken avidin. The DNA construct encoding scAvd contains four circularly permuted avidin domains, plus short linkers connecting the four domains into a single polypeptide chain.

Chicken genome analysis reveals novel genes encoding biotin-binding proteins related to avidin family.

Background: A chicken egg contains several biotin-binding proteins (BBPs), whose complete DNA and amino acid sequences are not known. In order to identify and characterise these genes and proteins we studied chicken cDNAs and genes available in the NCBI database and chicken genome database using the reported N-terminal amino acid sequences of chicken egg-yolk BBPs as search strings.

Results: Two separate hits showing significant homology for these N-terminal sequences were discovered.

A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo.

We have constructed a novel tetra-promoter vector (pBVboostFG) system that enables screening of gene/cDNA libraries for functional genomic studies. The vector enables an all-in-one strategy for gene expression in mammalian, bacterial and insect cells and is also suitable for direct use in vivo. Virus preparation is based on an improved mini Tn7 transpositional system allowing easy and fast production of recombinant baculoviruses with high diversity and negligible background.

Novel avidin-like protein from a root nodule symbiotic bacterium, Bradyrhizobium japonicum.

Bradyrhizobium japonicum is an important nitrogenfixing symbiotic bacterium, which can form root nodules on soybeans. These bacteria have a gene encoding a putative avidin- and streptavidin-like protein, which bears an amino acid sequence identity of only about 30% over the core regions with both of them. We produced this protein in Escherichia coli both as the full-length wild type and as a C-terminally truncated core form and showed that it is indeed a high affinity biotin-binding protein that resembles (strept)avidin structurally and functionally.

Design and construction of highly stable, protease-resistant chimeric avidins.

The chicken avidin gene family consists of avidin and seven separate avidin-related genes (AVRs) 1-7. Avidin protein is a widely used biochemical tool, whereas the other family members have only recently been produced as recombinant proteins and characterized. In our previous study, AVR4 was found to be the most stable biotin binding protein thus far characterized (T(m) = 106.