Predictors of disability in a childhood-onset systemic lupus erythematosus cohort: results from the CARRA Legacy Registry.

Objective Few descriptions of physical disability in childhood-onset SLE (cSLE) exist. We sought to describe disability in a large North American cohort of patients with cSLE and identify predictors of disability. Methods Sociodemographic and clinical data were obtained from the Childhood Arthritis and Rheumatology Research Alliance (CARRA) Legacy Registry for patients with cSLE enrolled between May 2010 and October 2014. The Childhood Health Assessment Questionnaire (CHAQ) was used to assess disability and physical functioning. Chi-square tests were used for univariate analyses, and multivariate logistic regression was used to assess predictors of disability. Results We analyzed data for 939 patients with cSLE. The median and mean CHAQ scores were 0 and 0.25, respectively, and 41% of the cohort had at least mild disability. Arthritis and higher pain scores were significantly associated with disability as compared to those without disability ( p < 0.001). In multivariate logistic regression analysis, low annual income, arthritis, and higher pain scores were associated with disability at baseline. Conclusions Disability as measured by baseline CHAQ was fairly common in cSLE patients in the CARRA Legacy Registry, and was associated with low household income, arthritis, and higher pain scores. In addition to optimal disease control, ensuring psychosocial supports and addressing pain may reduce disability in cSLE. Further study is needed of disability in cSLE.

Pig slurry acidification and separation techniques affect soil N and C turnover and N2O emissions from solid, liquid and biochar fractions.

The combined effects of pig slurry acidification, subsequent separation techniques and biochar production from the solid fraction on N mineralisation and N2O and CO2 emissions in soil were investigated in an incubation experiment. Acidification of pig slurry increased N availability from the separated solid fractions in soil, but did not affect N2O and CO2 emissions. However acidification reduced soil N and C turnover from the liquid fraction. The use of more advanced separation techniques (flocculation and drainage > decanting centrifuge > screw press) increased N mineralisation from acidified solid fractions, but also increased N2O and CO2 emissions in soil amended with the liquid fraction. Finally, the biochar production from the solid fraction of pig slurry resulted in a very recalcitrant material, which reduced N and C mineralisation in soil compared to the raw solid fractions.

Motives for adherence to a gluten-free diet: a qualitative investigation involving adults with coeliac disease.

BACKGROUND: Currently , the only treatment for coeliac disease is life long adherence to a strict gluten-free diet. Strict adherence to a gluten-free diet is challenging, with recent reports suggesting that adherence rates range from 42% to 91%. The present study aimed to: (i) identify motives for adhering to a gluten-free diet and (ii) explore factors implicated in adherence and non-adherence behaviour in terms of accidental and purposeful gluten consumption among adults with coeliac disease. METHODS: Two hundred and three adults with coeliac disease completed an online questionnaire. Using a qualitative design, relationships were examined between reported adherence and motivation to follow a gluten-free diet, as well as the onset, duration and severity of symptoms. RESULTS: Feelings of desperation ('hitting rock bottom') and needing to gain or lose weight were associated with the strictest adherence to a gluten-free diet. Participants who accidentally consumed gluten over the past week developed symptoms the most quickly and reported the most pain over the past 6 months. Participants who consumed gluten on purpose over the past week reported a shorter duration of symptoms and less pain over the past 6 months. CONCLUSIONS: Hitting rock bottom and needing to gain or lose weight were factors associated with the strictest adherence, when considered in the context of both accidental and purposeful gluten consumption. Future research is warranted to develop resources to help people with coeliac disease follow a strict gluten-free diet.

Hybrid carbon fiber/carbon nanotube composites for structural damping applications.

Carbon nanotubes (CNTs) were grown on the surface of carbon fibers utilizing a relatively low temperature synthesis technique; graphitic structures by design (GSD). To probe the effects of the synthesis protocols on the mechanical properties, other samples with surface grown CNTs were prepared using catalytic chemical vapor deposition (CCVD). The woven graphite fabrics were thermally shielded with a thin film of SiO2 and CNTs were grown on top of this film. Raman spectroscopy and electron microscopy revealed the grown species to be multi-walled carbon nanotubes (MWCNTs). The damping performance of the hybrid CNT-carbon fiber-reinforced epoxy composite was examined using dynamic mechanical analysis (DMA). Mechanical testing confirmed that the degradations in the strength and stiffness as a result of the GSD process are far less than those encountered through using the CCVD technique and yet are negligible compared to the reference samples. The DMA results indicated that, despite the minimal degradation in the storage modulus, the loss tangent (damping) for the hybrid composites utilizing GSD-grown MWCNTs improved by 56% compared to the reference samples (based on raw carbon fibers with no surface treatment or surface grown carbon nanotubes) over the frequency range 1-60 Hz. These results indicated that the energy dissipation in the GSD-grown MWCNTs composite can be primarily attributed to the frictional sliding at the nanotube/epoxy interface and to a lesser extent to the stiff thermal shielding SiO2 film on the fiber/matrix interface.

Mast cells protect against airway Mycoplasma pneumoniae under allergic conditions.

BACKGROUND: Infection with Mycoplasma pneumoniae (Mp) in asthma can occur both acutely and chronically with an associated Th2 inflammatory response and/or increased numbers of bronchial mast cells. Mast cells have previously been shown to promote mycoplasma clearance in mice; however, it is unknown whether mast cells would aid Mp clearance under allergic conditions. OBJECTIVE: Our aim was to determine the impact of allergic inflammation on mast cell-mediated lung Mp clearance. Furthermore, as we have previously demonstrated an essential role for IL-6 in lung Mp clearance we also investigated the role of mast cell-derived IL-6. METHODS: Mast cell-deficient (WBB6F1/J-Kit(W)/Kit(W-v)) mice were challenged with ovalbumin to induce airway inflammation before Mp infection. The role of mast cell-derived IL-6 in bacterial clearance was further investigated by reconstitution of mast cell-deficient mice with IL-6(-/-) mast cells. RESULTS: Allergic mast cell-deficient mice exhibited increased lung Mp burden compared with control littermates. Intravenous adoptive transfer of wild-type and IL-6(-/-) mast cells significantly improved Mp clearance in mast cell-deficient mice. Acutely after Mp infection, allergen-challenged mast cell-deficient mice had increased levels of the pro-inflammatory cytokines IL-6 and TNF-alpha in the bronchoalveolar lavage (BAL) fluid. The total number of neutrophils was also increased in mast cell-deficient mice. CONCLUSIONS: Our results establish that mast cells aid host defense against Mp in an allergic setting and that while IL-6 is necessary for lung Mp clearance, mast cell-derived IL-6 is not required.

Developmentally regulated expression of a Balbiani ring 1 gene for a 180-kD secretory polypeptide in Chironomus tentans salivary glands before larval/pupal ecdysis.

The expression of a Balbiani ring 1 gene that codes for a salivary gland-specific 180-kD secretory polypeptide (sp180) is regulated developmentally. Immunoblots of salivary gland protein incubated with an affinity-purified nonapeptide-reactive antibody demonstrated that the salivary gland content of sp180 increases as much as 10-fold between stages 8 and 10 of the fourth larval instar. Hybridization of RNA dot-blots with an oligonucleotide probe indicated that the observed increase in sp180 was preceded by a parallel 20-fold increase in the steady state level of its mRNA beginning between stages 7 and 8. In vitro nuclear transcription experiments demonstrated that there was a 10-fold acceleration in the rate of sp180 gene transcription between stages 6 and 10. The limited period of expression of the sp180 gene contrasted dramatically with the expression of Balbiani ring genes BR1, BR2 alpha, BR2 beta, and BR6, which code for the sp-I family of fibrous secretory polypeptides. The appearance of sp180 in secretion coincided with microscopically visible changes in the bundling of these fibrous polypeptides. At the same time, we noticed changes in the appearance and consistency of feeding tubes that larvae construct with this secretion. These results lead us to propose that sp180 may modify the structure or utilization of fibrous secretory polypeptides specifically for the assembly of pupation tubes necessary for larval/pupal ecdysis.

A novel giant secretion polypeptide in Chironomus salivary glands: implications for another Balbiani ring gene.

Chironomus salivary glands contain a family of high Mr (approximately 1,000 X 10(3)) secretion polypeptides thought to consist of three components: sp-Ia, sp-Ib, and sp-Ic. The use of a new extraction protocol revealed a novel high Mr component, sp-Id. Results of a survey of individual salivary glands indicated that sp-Id was widespread in more than a dozen strains of C. tentans and C. pallidivittatus. Sp-Id was phosphorylated at Ser residues, and a comparison of cyanogen bromide and tryptic peptide maps of 32P-labeled polypeptides suggested that sp-Ia, sp-Ib, and sp-Id are comprised of similar but nonidentical tandemly repeated amino acid sequences. We concluded that sp-Id is encoded by an mRNA whose size and nucleotide sequence organization are similar to Balbiani ring (BR) mRNAs that code for the other sp-I components. Furthermore, parallel repression of sp-Ib and sp-Id synthesis by galactose led us to hypothesize that both of their genes exist within Balbiani ring 2.

Frog perspective on the morphological difference between humans and chimpanzees.

The body shapes of humans and chimpanzees were compared quantitatively by criteria chosen for their capacity to discriminate well among the body shapes of frogs. By these criteria, the difference in body shape between humans and chimpanzees was found to be greater than that between the most dissimilar pairs of frogs examined--that is, frogs classified in separate taxonomic suborders. Even though the morphological diffference between the two primates is large by frog standards, the biochemical differences between the structural genes of these two species are small. The results of this study give quantitative support to the proposal that morphological evolution and biochemical evolution in structural genes can proceed at independent rates.

Nitrogen and phosphorus release from organic wastes and suitability as bio-based fertilizers in a circular economy.

The drive to a more circular economy has created increasing interest in recycling organic wastes as bio-based fertilizers. This study screened 15 different manures, digestates, sludges, composts, industry by-products, and struvites. Nitrogen (N) and phosphorous (P) release was compared following addition to soil. Three waste materials were then 'upgraded' using heating and pressure (105°C at 220 kPa), alkalinization (pH 10), or sonification to modify N and P release properties, and compared in a second soil incubation. Generally, maximum N release was negatively correlated with the CN ratio of the material (r = -0.6). Composted, dried, or raw organic waste materials released less N (mean of 10.8 ± 0.5%, 45.3 ± 7.2%, and 47.4 ± 3.2% of total N added respectively) than digestates, industry-derived organic fertilizer products, and struvites (mean of 58.2 ± 2.8%, 77.7 ± 6.0%, and 100.0 ± 13.1% of total N added respectively). No analyzed chemical property or processing type could explain differences in P release. No single upgrading treatment consistently increased N or P release. However, for one raw biosolid, heating at a low temperature (105°C) with pressure did increase N release as a percentage of total N added to soil from 30% to 43%.

Secondary structure of synthetic peptides derived from the repeating unit of a giant secretory protein from Chironomus tentans.

The secretory proteins of Chironomus tentans larvae, which are used to construct underwater feeding and pupation tubes, assemble into complexes in vitro. Members of a family of 1000 kDa proteins, the spIs, appear to form the fibrous backbone of the assembled complexes. The spIs consist of a core of tandemly repeating units of 60 to 90 amino acids that can be subdivided into two regions: the subrepeat region, made up of short internal repeats, and the constant region, which lacks simple subrepeats. We have synthesized peptides representative of the constant and subrepeat regions of one of the spIs, and have examined their secondary structure using Fourier transform IR and CD spectroscopy. The IR spectrum of the constant peptide indicates that this peptide has alpha-helical regions and beta-turns. The CD spectrum confirms this. The IR spectrum of the subrepeat peptide is similar to that of the poly(Gly)II helix, and also may indicate the presence of beta-turns. The CD spectrum is consistent with this helical structure. Extrapolation of these results to intact spIs is in agreement with secondary structure prediction and modeling studies. Our results indicate that the alpha-helices and poly(Gly)II-like helices are not arranged as coiled-coils, which are often found in fibrous proteins. We suggest that these structural elements may be in an unusual arrangement in the spIs, organized as alternating alpha-helices and poly(Gly)II or collagen-like helices, interspersed with beta-turns.

Single-stranded regions in DNA isolated from different developmental stages of the sea urchin.

Long-term labeled sea urchin embryo (Strongylocentrotus purpuratus) DNAs were examined for size of recovered pieces, single-strandedness, and length of continuous double-stranded regions. Sizing on neutral sucrose gradients indicates that morula stage DNA sediments predominantly at 31 S, blastula stage DNA at 27 S, and gastrula stage DNA as a broad range of sizes of greater than 29 S. Treatment of [3H]thymidine-labeled DNA with Aspergillus oryzae S1 nuclease removes 19% of the 3H from morula stage DNA, 4% of the 3H from blastula stage DNA, and less than 0.1% of the 3H from gastrula stage DNA. Sedimentation of S1 nuclease treated [3H]DNAs on alkaline sucrose gradients indicates that in native morula stage DNA there is a nick or gap in one strand approximately every 9700 base pairs, in native blastula stage DNA about every 3300 base pairs, and very few nicks or gaps in native gastrula stage DNA.

Disulfide bonds in a recombinant protein modeled after a core repeat in an aquatic insect's silk protein.

We constructed a gene encoding rCAS, recombinant constant and subrepeat protein, modeled after tandem repeats found in the major silk proteins synthesized by aquatic larvae of the midge, Chironomus tentans. Bacterially synthesized rCAS was purified to near homogeneity and characterized by several biochemical and biophysical methods including amino-terminal sequencing, amino acid compositional analysis, sedimentation equilibrium ultracentrifugation, and mass spectrometry. Complementing these techniques with quantitative sulfhydryl assays, we discovered that the four cysteines present in rCAS form two intramolecular disulfide bonds. Mapping studies revealed that the disulfide bonds are heterogeneous. When reduced and denatured rCAS was allowed to refold and its disulfide bonding state monitored, it again adopted a conformation with two intramolecular disulfide bonds. The inherent ability of rCAS to quantitatively form two intramolecular disulfide bonds may reflect a previously unknown feature of the in vivo silk proteins from which it is derived.

Selective deletion of large segments of Balbiani ring DNA during molecular cloning.

Transcription units in Balbiani ring 1 (BR1) and Balbiani ring 2 (BR2) of Chironomus tenans salivary glands are comprised of about 40 kb of repetitive DNA sequences organized in a satellite-like array. Because of this sequence organization, it was possible to prepare 30 to 40-kb target DNA fragments for cloning by performing limit restriction endonuclease digestion of high-Mr genomic DNA. These fragments were ligated to cohesive termini of the linearized cosmid, pHC79, packaged in vitro, and used to transduce Escherichia coli. Alternatively, target fragments were randomly sheared to a mean length of 8-10 kb, annealed to plasmid pBR322 using homopolymeric tails, and used for bacterial transformation. Recombinant cosmids and plasmids generally contained inserts which were proportional to the length of target fragments used in ligation reactions. However, the subset of recombinants that hybridized to 32P-labeled 75S RNA (highly enriched in BR1 + BR2 sequences) had disproportionately smaller inserts. With the exception of one metastable clone with a 2.1-kb insert, all others had inserts of 0.8 or 0.4 kb. Similar results were obtained in host cells that were recA- or recBC-. The most likely conclusion is that repetitive BR sequences are highly unstable during replication in E. coli and are selectively deleted.

Balbiani ring 3 in Chironomus tentans encodes a 185-kDa secretory protein which is synthesized throughout the fourth larval instar.

We have continued to map and identify genes encoding a family of secretory proteins. These proteins are synthesized in larval salivary glands of the midge, Chironomus tentans, and assemble in vivo into insoluble silk-like threads. The genes for several secretory proteins exist in Balbiani rings (BRs) on salivary-gland polytene chromosomes. A randomly primed cDNA clone, designated pCt185, hybridized in situ to BR3 and was shown on Northern blots to originate from a salivary gland-specific 6-kb poly(A) + RNA. The partial cDNA sequence contained 483 nucleotides including one open reading frame (ORF) encoding 160 amino acids (aa). A striking feature of the ORF was the periodic distribution of cysteine residues (Cys-X-Cys-X-Cys-X6-Cys) which occurred approximately every 22 aa. A cDNA-encoded 18-aa sequence was selected for chemical peptide synthesis. When affinity-purified antipeptide antibodies were incubated with a Western blot containing salivary-gland proteins they reacted specifically with a 185-kDa secretory protein (sp185). Developmental studies showed that sp185 and its mRNA were present in salivary glands throughout the fourth larval instar. Thus sp185 and a family of 1000-kDa secretory proteins are encoded by a class of genes that are expressed throughout the fourth instar. This contrasts with the developmentally regulated expression of the sp140 and sp195 genes whose expression is maximal during the prepupal stages of larval development.

A peptide-reactive antibody to a Balbiani ring gene product: immunological evidence that a 6.5-kb RNA in Chironomus tentans salivary glands is mRNA for a 180-kDa nonfibrous component of larval secretion.

An immunological approach was utilized to demonstrate that a tissue-specific Balbiani ring (BR) transcript in Chironomus tentans is the mRNA for a secreted 180-kDa polypeptide. Balbiani ring 1 (BR1) on the polytene chromosome IV of larval salivary glands contains a gene comprised of tandemly duplicated nucleotide sequences that are transcribed into a salivary gland-specific, 6.5-kb poly(A)+RNA for which a partial cDNA sequence exists [Dreesen et al., J. Biol. Chem. 260 (1985) 11824-11830]. A nonapeptide was synthesized so that its amino acid sequence corresponded to an open reading frame in the cDNA. This peptide was used to raise rabbit polyclonal antisera and to purify the peptide-reactive antibody by affinity chromatography. The affinity-purified antibody bound specifically to a 180-kDa polypeptide on Western blots containing extracts of total salivary gland protein. Western blot analysis of microdissected cellular vs. lumenal fractions of salivary glands indicated that this 180-kDa polypeptide was primarily localized in the lumen. Consequently, this polypeptide was designated a secretory polypeptide (sp180). Finally, the peptide-reactive antibody was used to localize sp180 in a nonfibrous component of salivary gland secretion by indirect immunofluorescence microscopy.

Characterization of a cloned, moderately repeated sequence from Balbiani ring 2 in Chironomus tentans.

pCtBR2-1 is a recombinant plasmid with a 750-bp insert of Chironomus tentans genomic DNA. When pCtBR2-1 was hybridized in situ to salivary gland polytene chromosomes, it hybridized exclusively to Balbiani ring 2 (BR2), a giant chromosomal puff. It was also shown that the insert contained four tandemly repeated sequences that were delineated by HinfI sites which occurred every 190 bp. The purified insert reassociated to C. tentans DNA with a C0t1/2 = 0.48 indicating that the sequence was moderately repeated within the genome. Hybridization of radioactive pCtBR2-1 to nitrocellulose blots containing partial HinfI digests of genomic DNA revealed that the 190-bp repeats were organized into one or more blocks of 11 to 12 copies in tandem. Hybridization of the recombinant plasmid to limit digests of genomic DNA also demonstrated that repeated sequences in BR2 were not homogeneous. As much as 70% of BR2 appeared to be represented by a 26-kb HhaI-resistant core, while the remaining 30% may have HhaI sites at 190-bp intervals, similar to pCtBR2-1.

Structure and polymorphism of the Chironomus thummi gene encoding special lobe-specific silk protein, ssp160.

cDNA encoding Chironomus thummi ssp160 was used to isolate a genomic clone that hybridized in situ to band A2b on polytene chromosome IV, the site of the ssp160 gene. DNA sequencing, primer extension and gene/cDNA nucleotide sequence alignment revealed the gene contains six exons and five introns; 70% of ssp160 is encoded in exon 3. Variations between cDNA and gene sequences led to the design of a polymerase chain reaction, restriction fragment length polymorphism assay that was subsequently used to demonstrate the existence of polymorphic alleles whose distribution varied between geographically separated populations of larvae. The polymorphism is associated with codon deletions in a six-amino-acid repeat containing an N-linked glycosylation motif. These deletions may have resulted from slipped-strand mispairing during DNA replication.

Identification of divergent homologs of Chironomus tentans sp185 and its Balbiani ring 3 gene in Australasian species of Chironomus and Kiefferulus.

A 185-kDa silk protein (sp185) from Chironomus tentans, present in both larval and prepupal silks, contains a striking amino acid sequence motif, Cys-X-Cys-X-Cys, which occurs about every 22-26 residues. Homologous proteins have been found in Chironomus pallidivittatus (sp185) and Chironomus thummi (sp220), which apparently differ in size but are very similar in overall composition and sequence. While surveying Australasian species of Chironomus and Kefferulus we obtained evidence for immunologically related silk protein having similar size and amino acid composition, but noticeably less Cys. Interspecies in situ hybridization to polytene chromosomes with C. tentans and C. pallidivittatus cDNA probes indicated that each species had a related gene. One pair of C. tentans cDNA-derived primers enabled polymerase chain reaction amplification of a discrete fragment of this gene from Kiefferulus 'cornishi'. Preliminary sequence information for this fragment confirmed the presence of an encoded Cys-X-Cys-X-Cys motif in what appeared to be a similar protein region containing less Cys. We conclude that homologs of C. tentans sp185 and its gene have been identified which may contain significant deviations in structure. Once suitable libraries are available, probes described here will be useful for selecting cDNA and genomic clones for detailed study.

Search for a 33.9 MeV/c(2) neutral particle in pion decay

The E815 (NuTeV) neutrino experiment has performed a search for a 33. 9 MeV/c(2) weakly interacting neutral particle produced in pion decay. Such a particle may be responsible for an anomaly in the timing distribution of neutrino interactions in the KARMEN experiment. E815 has searched for this particle's decays in an instrumented decay region; no evidence for this particle was found. The search is sensitive to pion branching ratios as low as 10(-13).