Investigating non-inferiority of internet-delivered versus face-to-face cognitive behavioural therapy for insomnia (CBT-I): a randomised controlled trial (iSleep well).

BACKGROUND: Insomnia is a highly prevalent disorder associated with numerous adverse health outcomes. Cognitive behavioural therapy for insomnia (CBT-I) is recommended as first-line treatment by clinical guidelines but is accessible to only a minority of patients suffering from insomnia. Internet-delivered CBT-I (iCBT-I) could contribute to the widespread dissemination of this first-line treatment. As there is insufficient evidence regarding non-inferiority, this study directly aims to compare therapist-guided internet-delivered versus face-to-face CBT-I in terms of insomnia severity post-treatment. Furthermore, a health-economic evaluation will be conducted, and potential benefits and disadvantages of therapist-guided iCBT-I will be examined. METHODS: This study protocol describes a randomised controlled two-arm parallel-group non-inferiority trial comparing therapist-guided iCBT-I with face-to-face CBT-I in routine clinical care. A total of 422 patients with insomnia disorder will be randomised and treated at 16 study centres throughout Germany. Outcomes will be assessed at baseline, 10 weeks after randomisation (post), and 6 months after randomisation (follow-up). The primary outcome is insomnia severity measured using the Insomnia Severity Index. Secondary outcomes include depression-related symptoms, quality of life, fatigue, physical activity, daylight exposure, adverse events related to treatment, and a health-economic evaluation. Finally, potential moderator variables and several descriptive and exploratory outcomes will be assessed (e.g. benefits and disadvantages of internet-delivered treatment). DISCUSSION: The widespread implementation of CBT-I is a significant healthcare challenge. The non-inferiority of therapist-guided iCBT-I versus face-to-face CBT-I will be investigated in an adequately powered sample in routine clinical care, with the same therapeutic content and same level of therapist qualifications provided with both interventions. If this trial demonstrates the non-inferiority of therapist-guided iCBT-I, healthcare providers may be more confident recommending this treatment to their patients, contributing to the wider dissemination of CBT-I. TRIAL REGISTRATION: Trial registration number in the German Clinical Trials Register: DRKS00028153 ( https://drks.de/search/de/trial/DRKS00028153 ). Registered on 16th May 2023.

Selective laser fiber welding on woven polymer fabrics for biomedical applications.

Localized cartilage damage is a common problem for younger patients. This can heal, but often results in a painful condition that requires intervention. A welded-woven three-dimensional polymer fabric has been suggested as a suitable cartilage replacement because such materials closely match the mechanical properties of cartilage. However, such materials fare poorly when evaluated with respect to wear. A microscopic investigation of wear mechanisms showed that it is critical that the fibers not deflect laterally under a normal load. This observation led to the use of a new process for selective laser welding of the surface layers of three-dimensional fabrics in order to improve their wear resistance. Experimental evaluations were made in a pin-on-disc arrangement with a biomimetic loading. All materials used in the studies have previously been used in orthopedic devices or meet the requirements for United States Pharmacopeial Convention (USP) Class VI biocompatibility approval. The wear rates were significantly reduced and the lifespan of the fabrics was markedly improved due to surface welding, making this a viable option for cartilage replacement in vivo.

Improved mechanical properties of acrylic bone cement with short titanium fiber reinforcement.

Acrylic bone cements are widely used in total joint arthroplasties to grout the prosthesis to bone. The changes in the tensile properties and fracture toughness of polymethylmethacrylate (PMMA) bone cements obtained by the addition of control and heat treated short titanium fibers are studied. Heat treatment of titanium fibers is conducted to precipitate titania particles on the fiber surface to improve the biocompatibility of the metal. Control and heat treated short titanium fibers (250 micro long and 20 micro diameter) were used as reinforcements at 3 volume %. X-ray diffraction indicated the presence of a rutile form of titania due to the heat treatments. The tensile and fracture properties were improved by the addition of fibers. Bone cements reinforced with titanium fibers heated at 550 degrees C for 1 h followed by 800 degrees C for 30 minutes show the largest increase in fracture toughness along with the smallest changes in elastic modulus and needs to be further investigated.

Improved mechanical properties of acrylic bone cement with short titanium fiber reinforcement.

Acrylic bone cements are widely used in total joint arthroplasties to grout the prosthesis to bone. The changes in the tensile properties and fracture toughness of polymethylmethacrylate (PMMA) bone cements obtained by the addition of control and heat treated short titanium fibers are studied. Heat treatment of titanium fibers is conducted to precipitate titania particles on the fiber surface, which may improve the biocompatibility of the metal. Control (non-heat treated) and heat treated short titanium fibers (250 microm long and 20microm diameter) were used as reinforcements at 3 volume %. X-ray diffraction indicated the presence of a rutile form of titania due to the heat treatments. Results indicate that the tensile and fracture properties of unfilled bone cement were improved by the addition of control and heat-treated fibers. The fracture properties of bone cements reinforced with control titanium fibers were at least 10% higher than those reinforced with heat treated titanium fibers. Therefore, we recommend further studies on the use of non-heat treated titanium fibers to reinforce acrylic bone cement.

Adhesion enhancement of steel fibers to acrylic bone cement through a silane coupling agent.

The use of a silane coupling agent (methacryloxypropyl-trichlorosilane) to improve the mechanical properties of steel fiber-reinforced acrylic bone cements was assessed. Changes to the tensile and fracture properties of bone cements reinforced with silane-coated or uncoated 316L stainless steel fibers of different aspect ratios were studied. Contact-angle measurements indicated that the coupling agent coats the metal surface through room temperature treatments in a short time (within 2 h). Push-out tests indicated that the interfacial shear strength of silane-coated 316L stainless steel rods is 141% higher than the uncoated rods. The elastic moduli, ultimate stresses, and fracture toughness of the silane-coated, steel fiber-reinforced bone cements are significantly higher than the bone cements reinforced with uncoated steel fibers. There were no differences in the tensile mechanical properties of the silane-coated or uncoated, steel fiber-reinforced cements after aging in a physiological saline solution, indicating that the bonding effectiveness is decreased by the intrusion of water at the metal-polymer interface. Because of possible biocompatibility issues with leaching of the silane coupling agent and no long-term mechanical benefit in simulated aging experiments, the use of these agents is not recommended for in vivo use.

Fracture toughness of steel-fiber-reinforced bone cement.

Fractures in the bone-cement mantle (polymethyl methacrylate) have been linked to the failure of cemented total joint prostheses. The heat generated by the curing bone cement has also been implicated in the necrosis of surrounding bone tissue, leading to loosening of the implants. The addition of reinforcements may improve the fracture properties of bone cement and decrease the peak temperatures during curing. This study investigates the changes in the fracture properties and the temperatures generated in the ASTM F451 tests by the addition of 316L stainless steel fibers to bone cement. The influence of filler volume fraction (5-15% by volume) and aspect ratios (19, 46, 57) on the fracture toughness of the acrylic bone cement was assessed. Increasing the volume fraction of the steel fibers resulted in significant increases in the fracture toughness of the steel-fiber-reinforced composite. Fracture-toughness increases of up to 2.63 times the control values were obtained with the use of steel-fiber reinforcements. No clear trend in the fracture toughness was discerned for increasing aspect ratios of the reinforcements. There is a decrease in the peak temperatures reached during the curing of the steel-fiber-reinforced bone cement, though the decrease is too small to be clinically relevant. Large increases in the fatigue life of acrylic bone cement were also obtained by the addition of steel fibers. These results indicate that the use of steel fibers may enhance the durability of cemented joint prostheses.

Bacterial expression and purification of recombinant Plasmodium yoelii circumsporozoite protein.

We report the expression and purification of recombinant rodent malarial Plasmodium yoelii circumsporozoite surface protein (PyCSP) in Escherichia coli. To facilitate purification of the recombinant protein, the PyCSP was expressed as an amino-terminal fusion protein to glutathione S-transferase and as a carboxy-terminal fusion protein to a hexahistidyl tag. The expression of the fusion protein was controlled by the inducible tac promoter. Under optimal conditions the immunoreactive PyCSP represented approximately 0.04% of the total cell lysate. Western blot analysis probing with an anti-PyCSP antibody revealed a wide array of immunoreactive bands. Material isolated by affinity purification on glutathione-Sepharose 4B resin also contained multiple bands indicative of premature termination or carboxyl-terminal degradation. Analysis of protein retained on a nickel nitrilotriacetic acid resin revealed evidence of amino-terminal deleted material. Combining the two mild affinity purifications resulted in isolation of a single immunoreactive protein of approximate molecular weight of 96 kDa. We anticipate that the approach described in this study will facilitate the production of highly purified recombinant proteins.

The translation initiation factor eIF4A from Schizosaccharomyces pombe is closely related to its mammalian counterpart.

We have isolated a cDNA clone encoding eIF4A from Schizosaccharomyces pombe. The deduced protein sequence is similar in length and sequence to other eIF4A proteins and exhibits highest similarity with the mammalian eIF4A protein. Hybridization with genomic DNA reveals two eIF4A genes located on two different chromosomes.

ATP hydrolysis by initiation factor 4A is required for translation initiation in Saccharomyces cerevisiae.

Saccharomyces cerevisiae translation initiation factor eIF-4A, an RNA helicase of the Asp-Glu-Ala-Asp (DEAD) box protein family, was mutated in the putative ATP binding site and expressed in Escherichia coli. Mutant proteins with alanine at position 66 replaced by glycine [eIF-4A(A66G)] or valine [eIF-4A(A66V)] were purified from Escherichia coli extracts and analyzed in vitro for activity in ATP crosslinking, ATP hydrolysis, RNA helicase, and translation assays. The results show that in vitro ATP hydrolysis activity, RNA helicase activity, and translation activity of eIF-4A correlate with in vivo activity of the factor. Whereas eIF-4A(A66G) showed wild-type activity in all assays, eIF-4A(A66V) was active in ATP crosslinking but inactive in ATP hydrolysis and RNA helicase assays. In vitro translation was supported by wild-type eIF-4A and eIF-4A(A66G) but not by eIF-4A(A66V). The results show that, for their translation, the majority of mRNAs from Saccharomyces cerevisiae including an mRNA with the initiator AUG positioned 8 nucleotides downstream of the cap structure require eIF-4A that is able to hydrolyze ATP.

D-E-A-D protein family of putative RNA helicases.

RNA metabolism plays a central role in cell growth. It is essential to regulate RNA synthesis, processing, stability and degradation. Conformational changes in RNA are key elements in regulating cellular processes. Recently, an increasing number of putative RNA helicases from different organisms ranging from Escherichia coli to humans and viruses have been identified. They are involved in diverse cellular functions such as RNA splicing, ribosome assembly, initiation of translation, spermatogenesis, embryogenesis, and cell growth and division. Based on sequence homologies these proteins were grouped in a family, the D-E-A-D box protein family (D-E-A-D = Asp-Glu-Ala-Asp). Some of the better characterized members have been shown to possess ATP-binding and hydrolysing activities as well as ATP-dependent RNA helicase activities. Most of the genes encoding such proteins have been isolated from yeast, on which we will focus in this review. From sequence data, three of the members form a subfamily, the D-E-A-H subfamily.

Translation initiation factor 4A from Saccharomyces cerevisiae: analysis of residues conserved in the D-E-A-D family of RNA helicases.

The eukaryotic translation initiation factor 4A (eIF-4A) possesses an in vitro helicase activity that allows the unwinding of double-stranded RNA. This activity is dependent on ATP hydrolysis and the presence of another translation initiation factor, eIF-4B. These two initiation factors are thought to unwind mRNA secondary structures in preparation for ribosome binding and initiation of translation. To further characterize the function of eIF-4A in cellular translation and its interaction with other elements of the translation machinery, we have isolated mutations in the TIF1 and TIF2 genes encoding eIF-4A in Saccharomyces cerevisiae. We show that three highly conserved domains of the D-E-A-D protein family, encoding eIF-4A and other RNA helicases, are essential for protein function. Only in rare cases could we make a conservative substitution without affecting cell growth. The mutants show a clear correlation between their growth and in vivo translation rates. One mutation that results in a temperature-sensitive phenotype reveals an immediate decrease in translation activity following a shift to the nonpermissive temperature. These in vivo results confirm previous in vitro data demonstrating an absolute dependence of translation on the TIF1 and TIF2 gene products.

Expression of translation initiation factor 4A from yeast and mouse in Saccharomyces cerevisiae.

The eukaryotic translation initiation factor 4A (eIF-4A) plays an important role in regulating initiation. To analyze its function in yeast, we carried out a mutational analysis of the TIF1 and TIF2 genes, which encode eIF-4A. Expression of these two yeast genes has also been investigated at the transcriptional level and it has been found that both are expressed in wild-type yeast cells. Analysis of the expression of eIF-4A-beta-galactosidase fusion proteins reveals that the TIF2 gene is more highly expressed than the TIF1 gene. Interestingly, the yeast eIF-4A protein shows a high degree of amino acid sequence similarity to the mouse homologue. However, we find that the mammalian factor does not support protein synthesis in yeast either in vivo or in vitro.

Translation in Saccharomyces cerevisiae: initiation factor 4A-dependent cell-free system.

Yeast Saccharomyces cerevisiae genes TIF1 and TIF2 (translation initiation factor) encode a protein tentatively called translation initiation factor (Tif) due to the similarity of its amino acid sequence and its molecular weight to mammalian eukaryotic initiation factor 4A. To clarify whether Tif is involved in translation, we produced an affinity-purified anti-Tif antibody by using Tif isolated from a Tif-overproducing yeast strain as immunogen and an Escherichia coli strain expressing Tif from an expression vector to provide the extract for affinity purification of the antibody. By using chromatographic procedures and the affinity-purified anti-Tif antibody as probe to identify Tif-containing fractions, we purified Tif from wild-type yeast cells. When yeast cells containing the only TIF1 gene on a plasmid under the control of the galactose-inducible CYC1-GAL10 promoter were grown in medium containing glucose as the carbon source, the production of Tif was shut off and growth was arrested. Lysates made from these cells were inactive in in vitro translation. Addition of Tif to these lysates restored in vitro protein synthesis. These results show that Tif is a translation factor, the yeast homologue of mammalian translation initiation factor 4A.

Deletion of the amino-terminal domain of asialoglycoprotein receptor H1 allows cleavage of the internal signal sequence.

Human asialoglycoprotein receptor H1 is a single-spanning membrane protein with an amino-terminal domain of 40 residues exposed to the cytoplasm and the carboxyl-terminal domain translocated to the exoplasmic side of the membrane. It has been shown earlier that the transmembrane segment functions as an internal uncleaved signal sequence for insertion into the endoplasmic reticulum. In a deletion protein lacking almost the entire cytoplasmic domain, the signal sequence is cleaved at the carboxyl-terminal end of the transmembrane segment. All available criteria suggest that the protein is processed by signal peptidase. The cytoplasmic domain of the receptor does not directly inhibit signal cleavage since it does not detectably hinder cleavage of the normally amino-terminal signal sequence of influenza hemagglutinin in fusion proteins. We suggest that by its size or structure it affects the position of the receptor in the membrane and thus the accessibility of the potential cleavage site to signal peptidase.