Depth-wise progression of osteoarthritis in human articular cartilage: investigation of composition, structure and biomechanics

SummaryObjectiveOsteoarthritis (OA) is characterized by the changes in structure and composition of articular cartilage. However, it is not fully known, what is the depth-wise change in two major components of the cartilage solid matrix, i.e., collagen and proteoglycans (PGs), during OA progression. Further, it is unknown how the depth-wise changes affect local tissue strains during compression. Our aim was to address these issues.MethodsData from the previous microscopic and biochemical measurements of the collagen content, distribution and orientation, PG content and distribution, water content and histological grade of normal and degenerated human patellar articular cartilage (n=73) were reanalyzed in a depth-wise manner. Using this information, a composition-based finite element (FE) model was used to estimate tissue function solely based on its composition and structure.ResultsThe orientation angle of collagen fibrils in the superficial zone of cartilage was significantly less parallel to the surface (P<0.05) in samples with early degeneration than in healthy samples. Similarly, PG content was reduced in the superficial zone in early OA (P<0.05). However, collagen content decreased significantly only at the advanced stage of OA (P<0.05). The composition-based FE model showed that under a constant stress, local tissue strains increased as OA progressed.ConclusionFor the first time, depth-wise point-by-point statistical comparisons of structure and composition of human articular cartilage were conducted. The present results indicated that early OA is primarily characterized by the changes in collagen orientation and PG content in the superficial zone, while collagen content does not change until OA has progressed to its late stage. Our simulation results suggest that impact loads in OA joint could create a risk for tissue failure and cell death

Promoting Parents' Use of Non-Pharmacological Methods and Assessment of Children's Postoperative Pain at Home

Background: Parents have reported challenges in assessing their child's postoperative pain at home.Aims: The purpose of this study was to evaluate the usefulness of the parental use of the Parents' Postoperative Pain Measure -tool (PPPM) on 1-3 -year-old children's non-pharmacological pain alleviation at home.Methodology: This was a non-randomized, prospective study with two parallel groups, where the parents in the intervention group were provided with PPPM in addition to a pain diary consisting of a verbal pain scale. The data were collected from 50 parents whose children had undergone day surgery in three Finnish university hospitals between January 2006 and June 2007. Parents completed questionnaires consisting of background information, verbal pain rating scale and a sub-scale measuring parents' use of non-pharmacological methods in children's postoperative pain alleviation.Results: Most children had mild postoperative pain after discharge, but in some children pain was moderate or severe. Non-pharmacological interventions were used commonly for pain alleviation in both groups, including holding the child in lap, comforting the child and spending time with the child more than usual during the recovery period after discharge. However, the use of non-pharmacological pain alleviation methods was 15% more common in the intervention group than in the control group. Parents of the intervention group had carried the child (p=0.04) and used distraction (p=0.05) more commonly than parents in control group. No group differences were found in parental assessments of the helpfulness of non-pharmacological pain alleviation methods.Conclusions: Children's pain remains under-treated and their pain alleviation can be promoted by providing the parents pain assessment tools, such as PPPM, to be used at home. The results can be utilized to further improve children's pain alleviation. More parental education is needed to promote their skills to alleviate the child's pain. Further research of the usefulness of the PPPM using larger samples is needed

Tannins, flavonoids and stilbenes in extracts of African savanna woodland trees Terminalia brownii, Terminalia laxiflora and Anogeissus leiocarpus showing promising antibacterial potential

Terminalia laxiflora, Terminalia brownii and Anogeissus leiocarpus are used as decoctions, macerations, infusions and fumigations in East and West African traditional medicine for treatment of infectious diseases and their symptoms. Using this ethnopharmacological information as a guideline for our research and owing to the fact that these species have not been subjected to in depth antibacterial and phytochemical studies, thirty-nine extracts of various polarities of the stem bark, stem wood and roots were studied for growth inhibitory effects against the human pathogenic bacteria Staphylococcus epidermidis, Staphylococcus aureus, Micrococcus luteus and Pseudomonas aeruginosa. Our results indicate that the studied species contain antibacterial compounds of a wide range of polarities. All polar root extracts of T. laxiflora and various polar extracts of T. brownii roots, including hot water decoctions, gave broad-spectrum antibacterial effects and low MIC values of 39 mu g/ml. The main ellagitannins in an ethyl acetate extract of the root of T. laxiflora were found to be corilagin and its derivative and punicalagin. A methanol extract of the roots of T. brownii contained methyl-(S)-flavogallonate and its derivative as the main identified ellagitannins. Moreover, both Terminalia species were found to contain ellagic acid xylopyranoside and methyl ellagic acid xyloside and pure ellagic acid was present in T. brownii. Pure punicalagin did not give as low MIC as an ethyl acetate extract of the roots of T. laxiflora, containing punicalagin as one of its main compounds, although this ellagitannin totally inhibited the growth of S. aureus at 125 mu g/ml and P. aeruginosa at 500 mu g/ml. Similarly, pure ellagic and gallic acid gave higher MIC values than the methanolic root extract of T. brownii against S. aureus and P. aeruginosa. Moreover, a Sephadex LH-20 fraction of the methanolic extract of the roots of T. brownii, enrichedwithmethyl-(S)-flavogallonate and its isomer, gave higher MIC values than the crude methanolic extract. These results suggest that the polyphenols in the extracts might act synergistically with each other. A methanolic soxhlet extract of the roots of A. leiocarpus, containing ampelopsin, aromadendrin, taxifolin, pinosylvin and 4'-methylpinosylvin gave a low MIC value of 39 mu g/ml against all bacterial strains used in this investigation. Our results demonstrate that the roots, stem bark and stem wood of T. brownii, T. laxiflora and A. leiocarpus are rich sources of (new) antimicrobial compounds and justify the uses of these plants for treatment of infections in African traditional medicine.Peer reviewe

Highly Pathogenic H5N1 Influenza A Virus Spreads Efficiently in Human Primary Monocyte-Derived Macrophages and Dendritic Cells

Influenza A viruses cause recurrent epidemics and occasional global pandemics. Wild birds are the natural reservoir of influenza A virus from where the virus can be transmitted to poultry or to mammals including humans. Mortality among humans in the highly pathogenic avian influenza H5N1 virus infection is even 60%. Despite intense research, there are still open questions in the pathogenicity of the H5N1 virus in humans. To characterize the H5N1 virus infection in human monocyte-derived macrophages (M phi s) and dendritic cells (DCs), we used human isolates of highly pathogenic H5N1/2004 and H5N1/1997 and low pathogenic H7N9/2013 avian influenza viruses in comparison with a seasonal H3N2/1989 virus. We noticed that the H5N1 viruses have an overwhelming ability to replicate and spread in primary human immune cell cultures, and even the addition of trypsin did not equalize the infectivity of H7N9 or H3N2 viruses to the level seen with H5N1 virus. H5N1 virus stocks contained more often propagation-competent viruses than the H7N9 or H3N2 viruses. The data also showed that human DCs and M phi s maintain 1,000- and 10,000-fold increase in the production of infectious H5N1 virus, respectively. Both analyzed highly pathogenic H5N1 viruses showed multi-cycle infection in primary human DCs and M phi s, whereas the H3N2 and H7N9 viruses were incapable of spreading in immune cells. Interestingly, H5N1 virus was able to spread extremely efficiently despite the strong induction of antiviral interferon gene expression, which may in part explain the high pathogenicity of H5N1 virus infection in humans

Phenolic compounds and expression of 4CL genes in silver birch clones and Pt4CL1a lines

A small multigene family encodes 4-coumarate:CoA ligases (4CLs) catalyzing the CoA ligation of hydroxycinnamic acids, a branch point step directing metabolites to a flavonoid or monolignol pathway. In the present study, we examined the effect of antisense Populus tremuloides 4CL (Pt4CL1) to the lignin and soluble phenolic compound composition of silver birch (Betula pendula) Pt4CL1a lines in comparison with non-transgenic silver birch clones. The endogenous expression of silver birch 4CL genes was recorded in the stems and leaves and also in leaves that were mechanically injured. In one of the transgenic Pt4CL1a lines, the ratio of syringyl (S) and guaiacyl (G) lignin units was increased. Moreover, the transcript levels of putative silver birch 4CL gene (Bp4CL1) were reduced and contents of cinnamic acid derivatives altered. In the other two Pt4CL1a lines changes were detected in the level of individual phenolic compounds. However, considerable variation was found in the transcript levels of silver birch 4CLs as well as in the concentration of phenolic compounds among the transgenic lines and non-transgenic clones. Wounding induced the expression of Bp4CL1 and Bp4CL2 in leaves in all clones and transgenic lines, whereas the transcript levels of Bp4CL3 and Bp4CL4 remained unchanged. Moreover, minor changes were detected in the concentrations of phenolic compounds caused by wounding. As an overall trend the wounding decreased the flavonoid content in silver birches and increased the content of soluble condensed tannins. The results indicate that by reducing the Bp4CL1 transcript levels lignin composition could be modified. However, the alterations found among the Pt4CL1a lines and the non-transgenic clones were within the natural variation of silver birches, as shown in the present study by the clonal differences in the transcripts levels of 4CL genes, soluble phenolic compounds and condensed tannins

The role of individual and social variables in task performance.

This paper reports on a data-based study in which we explored - as part of a larger-scale British-Hungarian research project - the effects of a number of affective and social variables on foreign language (L2) learners’ engagement in oral argumentative tasks. The assumption underlying the investigation was that students’ verbal behaviour in oral task situations is partly determined by a number of non-linguistic and non-cognitive factors whose examination may constitute a potentially fruitful extension of existing task-based research paradigms. The independent variables in the study included various aspects of L2 motivation and several factors characterizing the learner groups the participating students were members of (such as group cohesiveness and intermember relations), as well as the learners’ L2 proficiency and ‘willingness to communicate’ in their L1. The dependent variables involved objective measures of the students’ language output in two oral argumentative tasks (one in the learners’ L1, the other in their L2): the quantity of speech and the number of turns produced by the speakers. The results provide insights into the interrelationship of the multiple variables determining the learners’ task engagement, and suggest a multi-level construct whereby some independent variables only come into force when certain conditions have been met

Comparison of quantification methods for the condensed tannin content of extracts of Pinus pinaster bark

Bark from Pinus Pinaster is an interesting source of polyphenolic natural compounds, that can be used successful1y as total or partial replacement of conventional phenolic resins. These compounds, among other applications, are used as adhesives in the wood agglomerate industry. In this kind of application some problems remain to be solved in order to obtain a Pine extract of commercial value. It is necessary to optimise tije extraction procedure and select a suitable method for the quantification of the tannin content of the bark. In order to study these problems, the tannin extraction from the Pine bark was tested with an alkaline solution (NaOH), and with a fractionation procedure based on a sequence of an organic (ethanol) and aqueous extraction. The phenolic content of each extract or fraction was evaluated by the FolinCiocalteu colorimetric assay for total phenols and two procedures using the Stiasny reaction: the gravimetric Stiasny method and the indirect colorimetric procedure that uses the Folin-Ciocalteu reagent to evaluate the total phenols present in the extract solution before and after it condenses with formaldehyde. The yield value when the alkaline extraction is used is substantially higher than the values obtained with organic or aqueous solutions. However, the selectivity of the process is low. In fact, it was found that the alkaline extract Formaldehyde Condensable Phenolic Material (FCPM) content represents 9596 % of the total phenols content of the extract but this fraction is only ~ 40 % of the total mass of extract. So, the alkaline extract is relatively poor in phenolic material, exhibiting a large variety of non-phenolic extractives. On the other end, ethanol provides a very rich phenolic extract, in which 96 % of total phenols are condensable with formaldehyde, but exhibits a relatively low extraction yield. The aqueous extract presents the lowest extraction yield with low content either in phenolic material as in FCPM, but, as most of the phenolics had already been extracted by the previous organic extraction, especially the low molecular weight fractions, this result was predictable

No evidence of autoimmunity to human OX1 or OX2 orexin receptors in Pandemrix-vaccinated narcoleptic children

Narcolepsy type 1, likely an immune-mediated disease, is characterized by excessive daytime sleepiness and cataplexy. The disease is strongly associated with human leukocyte antigen (HLA) DQB1∗06:02. A significant increase in the incidence of childhood and adolescent narcolepsy was observed after a vaccination campaign with AS03-adjuvanted Pandemrix influenza vaccine in Nordic and several other countries in 2010 and 2011. Previously, it has been suggested that a surface-exposed region of influenza A nucleoprotein, a structural component of the Pandemrix vaccine, shares amino acid residues with the first extracellular domain of the human OX2 orexin/hypocretin receptor eliciting the development of autoantibodies. Here, we analyzed, whether H1N1pdm09 infection or Pandemrix vaccination contributed to the development of autoantibodies to the orexin precursor protein or the OX1 or OX2 receptors. The analysis was based on the presence or absence of autoantibody responses against analyzed proteins. Entire OX1 and OX2 receptors or just their extracellular N-termini were transiently expressed in HuH7 cells to determine specific antibody responses in human sera. Based on our immunofluorescence analysis, none of the 56 Pandemrix-vaccinated narcoleptic patients, 28 patients who suffered from a laboratory-confirmed H1N1pdm09 infection or 19 Pandemrix-vaccinated controls showed specific autoantibody responses to prepro-orexin, orexin receptors or the isolated extracellular N-termini of orexin receptors. We also did not find any evidence for cell-mediated immunity against the N-terminal epitopes of OX2. Our findings do not support the hypothesis that the surface-exposed region of the influenza nucleoprotein A would elicit the development of an immune response against orexin receptors. © 2020 The AuthorsPeer reviewe

Comparative Analysis of Whole-Genome Sequences of Influenza A(H1N1)pdm09 Viruses Isolated from Hospitalized and Nonhospitalized Patients Identifies Missense Mutations That Might Be Associated with Patient Hospital Admissions in Finland during 2009 to 2014

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Metabolic biomarker profiling for identification of susceptibility to severe pneumonia and COVID-19 in the general population

Biomarkers of low-grade inflammation have been associated with susceptibility to a severe infectious disease course, even when measured prior to disease onset. We investigated whether metabolic biomarkers measured by nuclear magnetic resonance (NMR) spectroscopy could be associated with susceptibility to severe pneumonia (2507 hospitalised or fatal cases) and severe COVID-19 (652 hospitalised cases) in 105,146 generally healthy individuals from UK Biobank, with blood samples collected 2007-2010. The overall signature of metabolic biomarker associations was similar for the risk of severe pneumonia and severe COVID-19. A multi-biomarker score, comprised of 25 proteins, fatty acids, amino acids, and lipids, was associated equally strongly with enhanced susceptibility to severe COVID-19 (odds ratio 2.9 [95%CI 2.1-3.8] for highest vs lowest quintile) and severe pneumonia events occurring 7-11 years after blood sampling (2.6 [1.7-3.9]). However, the risk for severe pneumonia occurring during the first 2 years after blood sampling for people with elevated levels of the multi-biomarker score was over four times higher than for long-term risk (8.0 [4.1-15.6]). If these hypothesis generating findings on increased susceptibility to severe pneumonia during the first few years after blood sampling extend to severe COVID-19, metabolic biomarker profiling could potentially complement existing tools for identifying individuals at high risk. These results provide novel molecular understanding on how metabolic biomarkers reflect the susceptibility to severe COVID-19 and other infections in the general population.Molecular Epidemiolog