Identification of individuals from low template blood samples using whole transcriptome shotgun sequencing.

Biological trace samples consisting of very few cells pose a challenge to conventional forensic genetic DNA analysis. RNA may be an alternative to DNA when handling low template samples. Whereas each cell only contains two copies of an autosomal DNA segment, the transcriptome retains much of the genomic variation replicated in abundant RNA fragments. In this study, we describe the development of a prototype RNA-based SNP selection set for forensic human identification from low template samples (50 pg gDNA). Whole blood from a subset of the Danish population (41 individuals) and blood stains subjected to degradation at room temperature for up to two weeks were analysed by whole transcriptome shotgun sequencing. Concordance was determined by DNA genotyping with the Infinium Omni5-4 SNP chip. In the 100 protein-coding genes with the most reads, 5214 bi-allelic SNPs with gnomAD minor allele frequencies > 0.1 in the African/African American, East Asian, and (non-Finnish) European populations were identified. Of these, 24 SNPs in 21 genes passed screening in whole blood and degraded blood stains, with a resulting mean match probability of 4.5 ∙ 10-9. Additionally, ancestry informative SNPs and SNPs in genes useful for body fluid identification were identified in the transcriptome. Consequently, shotgun sequencing of RNA from low template samples may be used for a vast host of forensic genetics purposes, including simultaneous human and body fluid identification, leading to direct donor identification in the identified body fluid.

Evaluation of DNAmAge in paired fresh, frozen, and formalin-fixed paraffin-embedded heart tissues.

The continued development in methylome analysis has enabled a more precise assessment of DNA methylation, but treatment of target tissue prior to analysis may affect DNA analysis. Prediction of age based on methylation levels in the genome (DNAmAge) has gained much interest in disease predisposition (biological age estimation), but also in chronological donor age estimation in crime case samples. Various epigenetic clocks were designed to predict the age. However, it remains unknown how the storage of the tissues affects the DNAmAge estimation. In this study, we investigated the storage method impact of DNAmAge by the comparing the DNAmAge of the two commonly used storage methods, freezing and formalin-fixation and paraffin-embedding (FFPE) to DNAmAge of fresh tissue. This was carried out by comparing paired heart tissue samples of fresh tissue, samples stored by freezing and FFPE to chronological age and whole blood samples from the same individuals. Illumina EPIC beadchip array was used for methylation analysis and the DNAmAge was evaluated with the following epigenetic clocks: Horvath, Hannum, Levine, Horvath skin+blood clock (Horvath2), PedBE, Wu, BLUP, EN, and TL. We observed differences in DNAmAge among the storage conditions. FFPE samples showed a lower DNAmAge compared to that of frozen and fresh samples. Additionally, the DNAmAge of the heart tissue was lower than that of the whole blood and the chronological age. This highlights caution when evaluating DNAmAge for FFPE samples as the results were underestimated compared with fresh and frozen tissue samples. Furthermore, the study also emphasizes the need for a DNAmAge model based on heart tissue samples for an accurate age estimation.

Interaction-focused music therapy with cancer-affected children and their significant others: a randomized controlled feasibility study with subsequent intervention (INMUT).

BACKGROUND: Paediatric oncology/haematology patients and their families are confronted with a life-threatening situation for which music therapy can be a cross-linguistic field of action. The creative act of making music together offers the possibility to strengthen competences and make conflicts tangible. Besides its complementing of evidence-based biomedical care, there is little research on the feasibility and efficacy of interactive music therapy including the diagnosed child and their significant others. METHODS: We conducted an assessor blind, prospective, multicentric feasibility randomized controlled trial (RCT) with subsequent intervention. Including overall 52 child-significant other dyads, INMUT investigates interaction-focused music therapy with cancer-affected children and their significant others (INMUT-KB; n = 21) compared to music therapy only with the child (MUT-K; n = 21) and a wait-list group (WLG; n = 10). The measurement points include the screening for a cancer diagnosis, psychometric baseline (pre-T1), initial assessment (T1/T2), music therapy sessions (T3-T9), final assessment (T10), final psychometric evaluation (post-T10), and 3-month follow-up (cat-T11). Feasibility and acceptability of the (1) research methodology, (2) intervention and (3) estimation of effect sizes will be assessed using qualitative and quantitative data. The proposed primary outcome includes the parent-child interaction (APCI), and the proposed secondary outcomes refer to subjective goal achievement (GAS), quality of life (KINDL), system-related functional level (EXIS), psychosocial stress (BAS), psychosomatic complaints (SCL-9k), and resources (WIRF). We plan to investigate the efficacy of INMUT-KB and MUT-K post-intervention (post-T10) within the RCT design and at 3-month follow-up (cat-T11). DISCUSSION: This study will provide insights into the feasibility of INMUT and the final sample needed for a confirmatory RCT. We will reflect on successfully implemented study procedures and, if necessary, provide recommendations for changes considering the design, procedures, measures, and statistical analyses. The discussion will conclude with an evaluation whether a confirmatory RCT is worth the investment of future resources, including the calculated number of child-significant other dyads needed based on the efficacy trends derived from this feasibility study. TRIAL REGISTRATION: ClinicalTrials.gov: NCT05534282; date of registration: June 23, 2022.

Intraarticular treatment with integrin α10ß1-selected mesenchymal stem cells affects microRNA expression in experimental post-traumatic osteoarthritis in horses.

Osteoarthritis (OA) remains a major cause of lameness in horses, which leads to lost days of training and early retirement. Still, the underlying pathological processes are poorly understood. MicroRNAs (miRNAs) are small non-coding RNAs that serve as regulators of many biological processes including OA. Analysis of miRNA expression in diseased joint tissues such as cartilage and synovial membrane may help to elucidate OA pathology. Since integrin α10ß1-selected mesenchymal stem cell (integrin α10-MSC) have shown mitigating effect on equine OA we here investigated the effect of integrin α10-MSCs on miRNA expression. Cartilage and synovial membrane was harvested from the middle carpal joint of horses with experimentally induced, untreated OA, horses with experimentally induced OA treated with allogeneic adipose-derived MSCs selected for the marker integrin α10-MSCs, and from healthy control joints. miRNA expression in cartilage and synovial membrane was established by quantifying 70 pre-determined miRNAs by qPCR. Differential expression of the miRNAs was evaluated by comparing untreated OA and control, untreated OA and MSC-treated OA, and joints with high and low pathology score. A total of 60 miRNAs were successfully quantified in the cartilage samples and 55 miRNAs were quantified in the synovial membrane samples. In cartilage, miR-146a, miR-150 and miR-409 had significantly higher expression in untreated OA joints than in control joints. Expression of miR-125a-3p, miR-150, miR-200c, and miR-499-5p was significantly reduced in cartilage from MSC-treated OA joints compared to the untreated OA joints. Expression of miR-139-5p, miR-150, miR-182-5p, miR-200a, miR-378, miR-409-3p, and miR-7177b in articular cartilage reflected pathology score. Several of these miRNAs are known from research in human patients with OA and from murine OA models. Our study shows that these miRNAs are also differentially expressed in experimental equine OA, and that expression depends on OA severity. Moreover, MSC treatment, which resulted in less severe OA, also affected miRNA expression in cartilage.

Fair concordance between Google Trends and Danish ornithologists in the assessment of temporal trends in Danish bird populations highlights the informational value of big data.

The ongoing depletion of natural systems and associated biodiversity decline is of growing international concern. Climate change is expected to exacerbate anthropogenic impacts on wild populations. The scale of impact on ecosystems and ecosystem services will be determined by the impact on a multitude of species and functional groups, which due to their biology and numbers are difficult to monitor. The IPCC has argued that surveillance or monitoring is critical and proposed that monitoring systems should be developed, which not only track developments but also function as "early warning systems." Human populations are already generating large continuous datasets on multiple taxonomic groups through internet searches. These time series could in principle add substantially to current monitoring if they reflect true changes in the natural world. We here examined whether information on internet search frequencies delivered by the Danish population and captured by Google Trends (GT) appropriately informs on population trends in 106 common Danish bird species. We compared the internet search activity with independent equivalent population trend assessments from the Danish Ornithological Society (BirdLife Denmark/DOF). We find a fair concordance between the GT trends and the assessments by DOF. A substantial agreement can be obtained by omitting species without clear temporal trends. Our findings suggest that population trend proxies from internet search frequencies can be used to supplement existing wildlife population monitoring and to ask questions about an array of ecological phenomena, which potentially can be integrated into an early warning system for biodiversity under climate change.

Integrin α10ß1-Selected Mesenchymal Stem Cells Reduce Pain and Cartilage Degradation and Increase Immunomodulation in an Equine Osteoarthritis Model.

OBJECTIVE: Integrin α10ß1-selected mesenchymal stem cells (integrin α10-MSCs) have previously shown potential in treating cartilage damage and osteoarthritis (OA) in vitro and in animal models in vivo. The aim of this study was to further investigate disease-modifying effects of integrin α10-MSCs. DESIGN: OA was surgically induced in 17 horses. Eighteen days after surgery, horses received 2 × 107 integrin α10-MSCs intra-articularly or were left untreated. Lameness and response to carpal flexion was assessed weekly along with synovial fluid (SF) analysis. On day 52 after treatment, horses were euthanized, and carpi were evaluated by computed tomography (CT), MRI, histology, and for macroscopic pathology and integrin α10-MSCs were traced in the joint tissues. RESULTS: Lameness and response to carpal flexion significantly improved over time following integrin α10-MSC treatment. Treated horses had milder macroscopic cartilage pathology and lower cartilage histology scores than the untreated group. Prostaglandin E2 and interleukin-10 increased in the SF after integrin α10-MSC injection. Integrin α10-MSCs were found in SF from treated horses up to day 17 after treatment, and in the articular cartilage and subchondral bone from 5 of 8 treated horses after euthanasia at 52 days after treatment. The integrin α10-MSC injection did not cause joint flare. CONCLUSION: This study demonstrates that intra-articular (IA) injection of integrin α10-MSCs appears to be safe, alleviate pathological changes in the joint, and improve joint function in an equine post-traumatic osteoarthritis (PTOA) model. The results suggest that integrin α10-MSCs hold promise as a disease-modifying osteoarthritis drug (DMOAD).

Multi-Omic Temporal Landscape of Plasma and Synovial Fluid-Derived Extracellular Vesicles Using an Experimental Model of Equine Osteoarthritis.

Extracellular vesicles (EVs) contribute to osteoarthritis pathogenesis through their release into joint tissues and synovial fluid. Synovial fluid-derived EVs have the potential to be direct biomarkers in the causal pathway of disease but also enable understanding of their role in disease progression. Utilizing a temporal model of osteoarthritis, we defined the changes in matched synovial fluid and plasma-derived EV small non-coding RNA and protein cargo using sequencing and mass spectrometry. Data exploration included time series clustering, factor analysis and gene enrichment interrogation. Chondrocyte signalling was analysed using luciferase-based transcription factor activity assays. EV protein cargo appears to be more important during osteoarthritis progression than small non-coding RNAs. Cluster analysis revealed plasma-EVs represented a time-dependent response to osteoarthritis induction associated with supramolecular complexes. Clusters for synovial fluid-derived EVs were associated with initial osteoarthritis response and represented immune/inflammatory pathways. Factor analysis for plasma-derived EVs correlated with day post-induction and were primarily composed of proteins modulating lipid metabolism. Synovial fluid-derived EVs factors represented intermediate filament and supramolecular complexes reflecting tissue repair. There was a significant interaction between time and osteoarthritis for CRE, NFkB, SRE, SRF with a trend for osteoarthritis synovial fluid-derived EVs at later time points to have a more pronounced effect.

Fresh and frozen cardiac tissue are comparable in DNA methylation array ß-values, but formalin-fixed, paraffin-embedded tissue may overestimate DNA methylation levels.

Untreated fresh cardiac tissue is the optimal tissue material for investigating DNA methylation patterns of cardiac biology and diseases. However, fresh tissue is difficult to obtain. Therefore, tissue stored as frozen or formalin-fixed, paraffin-embedded (FFPE) is widely used for DNA methylation studies. It is unknown whether storage conditions alter the DNA methylation in cardiac tissue. In this study, we compared the DNA methylation patterns of fresh, frozen, and FFPE cardiac tissue to investigate if the storage method affected the DNA methylation results. We used the Infinium MethylationEPIC assay to obtain genome-wide methylation levels in fresh, frozen, and FFPE tissues from nine individuals. We found that the DNA methylation levels of 21.4% of the examined CpG sites were overestimated in the FFPE samples compared to that of fresh and frozen tissue, whereas 5.7% were underestimated. Duplicate analyses of the DNA methylation patterns showed high reproducibility (precision) for frozen and FFPE tissues. In conclusion, we found that frozen and FFPE tissues gave reproducible DNA methylation results and that frozen and fresh tissues gave similar results.

Biomechanical performance of a novel light-curable bone fixation technique.

Traumatic bone fractures are often debilitating injuries that may require surgical fixation to ensure sufficient healing. Currently, the most frequently used osteosynthesis materials are metal-based; however, in certain cases, such as complex comminuted osteoporotic fractures, they may not provide the best solution due to their rigid and non-customizable nature. In phalanx fractures in particular, metal plates have been shown to induce joint stiffness and soft tissue adhesions. A new osteosynthesis method using a light curable polymer composite has been developed. This method has demonstrated itself to be a versatile solution that can be shaped by surgeons in situ and has been shown to induce no soft tissue adhesions. In this study, the biomechanical performance of AdhFix was compared to conventional metal plates. The osteosyntheses were tested in seven different groups with varying loading modality (bending and torsion), osteotomy gap size, and fixation type and size in a sheep phalanx model. AdhFix demonstrated statistically higher stiffnesses in torsion (64.64 ± 9.27 and 114.08 ± 20.98 Nmm/° vs. 33.88 ± 3.10 Nmm/°) and in reduced fractures in bending (13.70 ± 2.75 Nm/mm vs. 8.69 ± 1.16 Nmm/°), while the metal plates were stiffer in unreduced fractures (7.44 ± 1.75 Nm/mm vs. 2.70 ± 0.72 Nmm/°). The metal plates withstood equivalent or significantly higher torques in torsion (534.28 ± 25.74 Nmm vs. 614.10 ± 118.44 and 414.82 ± 70.98 Nmm) and significantly higher bending moments (19.51 ± 2.24 and 22.72 ± 2.68 Nm vs. 5.38 ± 0.73 and 1.22 ± 0.30 Nm). This study illustrated that the AdhFix platform is a viable, customizable solution that is comparable to the mechanical properties of traditional metal plates within the range of physiological loading values reported in literature.

Use of admission serum neutrophil gelatinase-associated lipocalin (NGAL) concentrations as a marker of sepsis and outcome in neonatal foals.

BACKGROUND: Equine neonatal sepsis can be challenging to diagnose and prognosticate. Neutrophil gelatinase-associated lipocalin (NGAL), a new marker of renal damage and inflammation, can potentially be helpful. OBJECTIVES: To evaluate NGAL in neonatal foals with sepsis, and assess its relation to outcome. ANIMALS: Foals ≤ 14 days, with admission blood analysis and stored serum. METHODS: NGAL was measured on stored serum from 91 foals. Foals were scored for sepsis and survival and categorized according to sepsis status (septic, sick non-septic, healthy, and uncertain sepsis status) and outcome groups (survivors and non-survivors). The septic foals were further sub-categorized according to severity (normal sepsis, severe sepsis and septic shock). A Kruskal-Wallis test was used to compare serum NGAL concentrations in survivors and non-survivors, in the sepsis status groups, and in the sepsis severity groups. Optimal cut-off values for serum NGAL concentrations to diagnose sepsis and outcome were determined with receiver operating characteristic (ROC) curves. NGAL was compared to creatinine and SAA. RESULTS: Median serum NGAL concentrations were significantly higher in septic than non-septic foals. However, serum NGAL concentrations did not differ between sepsis severity subgroups. Serum NGAL concentrations were significantly lower in survivors than in non-survivors. Optimal cut-off values of serum NGAL concentrations were 455 µg/L (sensitivity 71.4%, specificity 100%) and 1104 µg/L (sensitivity 39.3%, specificity 95.2%) for predicting sepsis and non-survival, respectively. NGAL correlated to SAA, but not to creatinine. NGAL performed similarly to SAA to diagnose sepsis. CONCLUSION: Serum NGAL concentrations may be useful for diagnosing sepsis and predicting outcome.

Comparison of whole transcriptome sequencing of fresh, frozen, and formalin-fixed, paraffin-embedded cardiac tissue.

The use of fresh tissue for molecular studies is preferred but often impossible. Instead, frozen or formalin-fixed, paraffin-embedded (FFPE) tissues are widely used and constitute valuable resources for retrospective studies. We assessed the utility of cardiac tissue stored in different ways for gene expression analyses by whole transcriptome sequencing of paired fresh, frozen, and FFPE tissues. RNA extracted from FFPE was highly degraded. Sequencing of RNA from FFPE tissues yielded higher proportions of intronic and intergenic reads compared to RNA from fresh and frozen tissues. The global gene expression profiles varied with the storage conditions, particularly mitochondrial and long non-coding RNAs. However, we observed high correlations among protein-coding transcripts (ρ > 0.94) with the various storage conditions. We did not observe any significant storage effect on the allele-specific gene expression. However, FFPE had statistically significantly (p < 0.05) more discordant variant calls compared to fresh and frozen tissue. In conclusion, we found that frozen and FFPE tissues can be used for reliable gene expression analyses, provided that proper quality control is performed and caution regarding the technical variability is withheld.

DNA quality evaluation of formalin-fixed paraffin-embedded heart tissue for DNA methylation array analysis.

Archived formalin-fixed and paraffin-embedded (FFPE) heart tissue from autopsied individuals represents an important resource for investigating the DNA methylation of heart tissue of deceased individuals. The DNA quality of FFPE tissue from autopsies may be decreased, affecting the DNA methylation measurements. Therefore, inexpensive screening methods for estimating DNA quality are valuable. We investigated the correlation between the DNA quality of archived FFPE heart tissue examined with the Illumina Infinium HD FFPE QC assay (Infinium QC) and Thermo Fisher's Quantifiler Trio DNA Quantification kit (QuantifilerTrio), respectively, and the amount of usable DNA methylation data as measured by the probe detection rate (probe DR) obtained with the Illumina Infinium MethylationEPIC array. We observed a high correlation (r2 = 0.75; p < 10-11) between the QuantifilerTrio degradation index, DI, and the amount of usable DNA methylation data analysed with SeSAMe, whereas a much weaker correlation was observed between the Infinium QC and SeSAMe probe DR (r2 = 0.17; p < 0.05). Based on the results, QuantifilerTrio DI seems to predict the proportion of usable DNA methylation data analysed with the Illumina Infinium MethylationEPIC array and SeSAMe by a linear model: SeSAMe probe DR = 0.80-log10(DI) × 0.25.

Serum amyloid A does not predict non-survival in hospitalised adult horses with acute colitis.

BACKGROUND: Predicting non-survival in horses with acute colitis improves early decision making. Therefore, this study aimed to determine the prognostic value of serum amyloid A (SAA) and other clinicopathological and clinical variables in adult horses with acute colitis. METHODS: Clinical variables, SAA and other blood biomarkers, including plasma L-lactate (lactate), were assessed in 176 horses with acute colitis. A multivariate model for the prediction of non-survival was constructed. Icelandic horses were analysed separately. RESULTS: Admission SAA was similar in survivors (median 548 mg/L; range 0-5453 mg/L) and non-survivors (396 mg/L; 0-5294) (p = 0.43). A model for non-survival included year of admission, lactate, heart rate, age and colic duration of more than 24 hours. Icelandic horses had a relative risk of 2.9 (95% confidence interval = 2.2-3.8) for acute colitis compared to other breeds. Lactate in Icelandic horses was higher than that in other breeds in both survivors (4.0 mmol/L, range 1.0-12.7 vs. 2.0, 0.7-12.5) and non-survivors (10.0, 1.5-26 vs. 5.4, 0.8-22) (p < 0.001). LIMITATIONS: The prognostic value of repeated measurements of SAA could not be assessed in this study, as 71% of the non-surviving horses died within a day of admission. CONCLUSION: Admission SAA did not predict non-survival. Breed needs consideration when lactate is evaluated as a predictor for non-survival in horses with colitis.

Concentrations of neutrophil gelatinase-associated lipocalin are increased in serum and peritoneal fluid from horses with inflammatory abdominal disease and non-strangulating intestinal infarctions.

BACKGROUND: Neutrophil gelatinase-associated lipocalin (NGAL) is produced in response to inflammation in horses, but it has not yet been investigated as a biomarker in horses with the abdominal disease. OBJECTIVES: To investigate NGAL in serum and peritoneal fluid in horses with the abdominal disease. STUDY DESIGN: Retrospective clinical study. METHODS: Data from medical records of 270 horses admitted with acute abdominal disease (simple obstructions [n = 43], strangulating obstructions [n = 104], inflammatory abdominal disease [n = 99], non-strangulating intestinal infarction [NSII, n = 24]) and 9 healthy control horses were reviewed, and serum and peritoneal fluid samples were retrieved from a biobank. NGAL was measured in serum and peritoneal fluid by ELISA. Differences in NGAL concentrations between groups were assessed. RESULTS: Healthy horses had low serum and peritoneal fluid concentrations of NGAL (median = 21.0 and 9.5 µg/L, respectively). Neither serum nor peritoneal fluid NGAL concentrations (median serum, peritoneal fluid) differed between healthy horses and horses with simple (28.1 µg/L, 13.0 µg/L) and strangulating intestinal obstructions (34.7 µg/L, 38.4 µg/L). Horses with NSII (308.0 µg/L, 2163.0 µg/L) and inflammatory abdominal disease (171.1 µg/L, 314.1 µg/L) had higher serum and peritoneal fluid concentrations of NGAL than the other groups (p < 0.001). Peritoneal fluid NGAL concentrations in horses with NSII were higher than in horses with inflammatory abdominal disease (p = 0.03). MAIN LIMITATIONS: Retrospective study design; small group of control horses. CONCLUSIONS: NGAL is a marker of inflammation in horses with the abdominal disease. The high peritoneal fluid concentration of NGAL in horses with NSII compared to all other groups may render NGAL useful for identifying this condition, which is otherwise difficult to differentiate from other types of peritonitis. Thereby, NGAL may potentially facilitate timely surgical intervention in this group of patients. The results of this study must be evaluated in larger, and preferably prospective, studies to obtain a more comprehensive evaluation of the diagnostic utility of NGAL.

CONTEXTO: Lipocalina associada à gelatinase de neutrófilos (NGAL) é produzida em resposta à inflamação em equinos, mas ainda não foi investigada como biomarcador de equinos com doenças abdominais. OBJETIVOS: Investigar a NGAL no soro e líquido peritoneal (LP) de equinos com doenças abdominais. DELINEAMENTO DO ESTUDO: Estudo retrospectivo. METODOLOGIA: Informação de prontuários médicos de 270 equinos com doença abdominal (obstrução simples [n = 43], obstrução estrangulante [n = 104], doença abdominal inflamatória [n = 99], lesão de infarto não estrangulante [NSII, n = 24]) e nove equinos saudáveis como controle foram revisados, e o soro e LP foram obtidos de um biobank. NGAL foi mensurado no soro e LP através de ELISA. Diferenças na concentração de NGAL entre os grupos foi avaliada. RESULTADOS: Cavalos saudáveis tiveram concentração de NGAL baixas no soro e no LP (mediana = 21.0 e 9.5 µg/L, respectivamente). A concentração de NGAL no soro e no LP não diferiram entre cavalos saudáveis e cavalos com obstrução simples (28.1,13.0) e obstrução estrangulante (34.7, 38.4). Cavalos com NSII (308.0, 2163.0) e doença abdominal inflamatória (171.1314.1) tiveram concentrações mais altas de NGAL no soro e LP quando comparados aos outros grupos (p < 0.001). A concentração de NGAL no LP em cavalos com NSII foi mais alta do que em cavalos com doença abdominal inflamatória (p = 0.03). PRINCIPAIS LIMITAÇÕES: Delineamento de estudo retrospectivo e pequeno grupo de equinos controle. CONCLUSÕES: NGAL é um marcador de inflamação em cavalos com doença abdominal. A concentração alta de NGAL no LP de cavalos com NSII comparado com os outros grupos indica que NGAL pode ser útil em identificar essa condição, o que é usualmente difícil de diferenciar de outros tipos de peritonite. Contudo, NGAL tem o potencial de facilitar a decisão rápida da necessidade de intervenção cirúrgica nesse grupo de pacientes. Os resultados deste estudo devem ser avaliados em uma população maior (e preferencialmente prospectiva) para obter uma melhor avaliação compreensiva da utilidade diagnóstica do NGAL.

Differentiating Strongylus vulgaris-associated, non-strangulating intestinal infarctions from idiopathic peritonitis and acute colitis in horses.

BACKGROUND: Horses with non-strangulating intestinal infarction (NSII) are often misdiagnosed with idiopathic peritonitis or acute colitis. Early diagnosis is essential to ensure early surgical intervention and improve survival. METHODS: Clinical and laboratory data from horses admitted to the University of Copenhagen Large Animal Teaching Hospital with NSII, idiopathic peritonitis or acute colitis between 2009 and 2018 were used for univariate comparisons and a multivariable logistic regression model for prediction of NSII. RESULTS: Two hundred and thirty-one horses were included. A multivariable model for the prediction of NSII included gastric reflux (more than 5 L) (odds ratio [OR] 8.7; 95% confidence interval [CI] 2.1-36.2), abnormal findings palpated per rectum (intestinal dilatations/impactions [OR 4.43; 95% CI 1.43-13.38], colon displacements [OR 23.16; 95% CI 5.26-101.97] or intestinal mass [OR 179.7; 95% CI 23.5-1375.5]), white blood cell count (OR 1.2; 95% CI 1.1-1.4), packed cell volume (OR 0.9; 95% CI 0.8-0.9), age (OR 0.9; 95% CI 0.8-1.0) and heart rate (OR 1.1; 95% CI 1.0-1.1). The model had a low false positive rate (5%), but a high false negative rate (50%). LIMITATIONS: Due to the retrospective nature of the study, sample collection was inconsistent, resulting in missing values. CONCLUSION: The model had some capability in predicting NSII. However, the high risk of false negatives means that exploratory laparotomy should be considered in horses with peritonitis of unknown aetiology in areas where Strongylus vulgaris is prevalent and occurrence of idiopathic peritonitis is low.

Use of serum amyloid A in equine medicine and surgery.

Serum amyloid A (SAA) has become an indispensable part of the management of equine patients in general practice and specialized hospital settings. Although several proteins possess acute phase properties in horses, the usefulness of SAA exceeds that of other acute phase proteins. This is due to the highly desirable kinetics of the equine SAA response. SAA concentrations exhibit a rapid and pronounced increase in response to inflammation and a rapid decline after the resolution of inflammation. This facilitates the detection of inflammatory disease and real-time monitoring of inflammatory activity. SAA may be used in all stages of patient management: (1) before diagnosis (to rule in/rule out inflammatory disease), (2) at the time of diagnosis (to assess the severity of inflammation and assist in prognostication), and (3) after diagnosis (to monitor changes in inflammatory activity in response to therapy, with relapse of disease, or with infectious/inflammatory complications). By assessing other acute phase reactants in addition to SAA, clinicians can succinctly stage inflammation. White blood cell counts and serum iron concentration change within hours of an inflammatory insult, SAA within a day, and fibrinogen within 2-3 days; the interrelationship of these markers thus indicates the duration and activity of the inflammatory condition. Much research on the equine SAA response and clinical use has been conducted in the last decade. This is the prerequisite for the evidence-based use of this analyte. However, still today, most published studies involve a fairly low number of horses. To obtain solid evidence for use of SAA, future studies should be designed with larger sample sizes.

Psychometric properties of the Marschak Interaction Method of Psychometrics and the Assessment of Parent-Child Interaction within residential care and non-referred settings.

Introduction: Assessment and identification of children with developmental needs and their interaction with primary caregivers are critical for emotional and social development. However, to the best of our knowledge, there is a scarcity of valid observation-based tools that guide the work with family communication, which is essential for the child's healthy development. Method: The Marschak Interaction Method of Psychometrics (MIM-P) and Assessment of Parent-Child Interaction (APCI) are both interaction and observation-based assessment tool, and they were explored for their validity and reliability in assessing caregiver-child interaction. The study included 30 trained and certified professionals who recruited referred and non-referred caregiver-child dyads over 11 months. Assessment data was collected from 139 caregiver-child dyads for the MIM-P with 278 individuals (100 referred and 178 non-referred) and 129 caregiver-child dyads for the APCI with 257 individuals (95 referred and 162 non-referred). Results: The psychometric analyses show that both the MIM-P and APCI presents relevant sources of reliability and validity for assessing caregiver-child interaction including interrater reliability, internal consistency, test re-test reliability as well as concurrent and construct validity. Discussion and conclusion: The study highlights the need for observation-based assessment tools within social work and contributes to the understanding of the importance of relationships and interaction in children's emotional and social development. However, further research is needed to explore norms and further strengthen implementation and quality of the tools.

Temporal extracellular vesicle protein changes following intraarticular treatment with integrin α10ß1-selected mesenchymal stem cells in equine osteoarthritis.

Introduction: Equine osteoarthritis (OA) is a heterogeneous, degenerative disease of the musculoskeletal system with multifactorial causation, characterized by a joint metabolic imbalance. Extracellular vesicles are nanoparticles involved in intracellular communication. Mesenchymal stem cell (MSC) therapy is a form of regenerative medicine that utilizes their properties to repair damaged tissues. Despite its wide use in veterinary practice, the exact mechanism of action of MSCs is not fully understood. The aim of this study was to determine the synovial fluid extracellular vesicle protein cargo following integrin α10ß1-selected mesenchymal stem cell (integrin α10-MSC) treatment in an experimental model of equine osteoarthritis with longitudinal sampling. Methods: Adipose tissue derived, integrin α10-MSCs were injected intraarticularly in six horses 18 days after experimental induction of OA. Synovial fluid samples were collected at day 0, 18, 21, 28, 35, and 70. Synovial fluid was processed and extracellular vesicles were isolated and characterized. Extracellular vesicle cargo was then analyzed using data independent acquisition mass spectrometry proteomics. Results: A total of 442 proteins were identified across all samples, with 48 proteins differentially expressed (FDR ≤ 0.05) between sham-operated control joint without MSC treatment and OA joint treated with MSCs. The most significant pathways following functional enrichment analysis of the differentially abundant protein dataset were serine endopeptidase activity (p = 0.023), complement activation (classical pathway) (p = 0.023), and collagen containing extracellular matrix (p = 0.034). Due to the lack of an OA group without MSC treatment, findings cannot be directly correlated to only MSCs. Discussion: To date this is the first study to quantify the global extracellular vesicle proteome in synovial fluid following MSC treatment of osteoarthritis. Changes in the proteome of the synovial fluid-derived EVs following MSC injection suggest EVs may play a role in mediating the effect of cell therapy through altered joint homeostasis. This is an important step toward understanding the potential therapeutic mechanisms of MSC therapy, ultimately enabling the improvement of therapeutic efficacy.

Music, families and interaction (MUFASA): a protocol article for an RCT study.

BACKGROUND: General mental health and interpersonal skills of families are crucial to children's development and future. Research suggests a link between participation in music activities and individuals' own sense of mental health, as well as an effect in objective measures of health such as quality of life, social skills, and rehabilitation of various kinds. However, in Denmark there are not many services for families with school-aged children (7-12 years) that focus on prevention and strengthening family interaction hereby minimising the risk of children not thriving and developing optimally in terms of social and emotional skills and competences. The purpose of this study is to investigate the effect of shared music activities on family interaction, parental stress, and child well-being. METHODS: The study is a controlled effect study where 40 family dyads from Aalborg and Aarhus are randomized into 4 different groups including music therapy activities, community music, family concerts and a control group. Measurements of family interaction (Assessment of Parent Child Interaction, APCI) and mental health (Parental Stress Scale, PSS, and the Strengths and Difficulties Questionnaire, SDQ) will be made at baseline/pre intervention, at post intervention and a follow-up measurement 12 month after baseline (month 1, 3 and 12) 3 times in total. Furthermore, data about the family dyad's regular participation in music events as part of every-day life at the same measure points (month 1, 3 and 12). Since this is a planned prospective study, results are not yet available, but clinical experience from a feasibility study in 2021 were promising and expected challenges and changes are discussed in the article. DISCUSSION: Findings of this study will be relevant for all fields where music is applied for families, such as education, mental health, social work and for basic research on the study of music and interaction. Music used as a resource for families is crucial in understanding how different modalities can influence the family interaction including emotional communication and attachment. TRIAL REGISTRATION: ISRCTN, ISRCTN17290015, Registered 03 March 2022, https://www.isrctn.com/ISRCTN17290015 DATE AND PROTOCOL VERSION: July 2022, version 1. Protocol is planned to be updated after finalized recruitment during second data collection point and again after the third and last data collection point (see Additional file 1: SPIRIT Checklist).