Hevosten trakealima- ja keuhkohuuhtelunäytteet yliopistollisessa hevossairaalassa

Hengitystiesairauteen viittaava oireilu on yksi yleisimmistä syistä hevosen tutkimiselle. Erilaiset hengitysteistä otetut näytteet helpottavat diagnoosin tekoa. Jo pitkään on yritetty selvittää, kumpi näytteenottomenetelmä, trakealima- vai keuhkohuutelunäyteenotto, olisi diagnostisempi. Useimmissa tutkimuksissa on kuitenkin päädytty siihen, että diagnostisinta olisi ottaa molemmat näytteet. Tässä lisensiaatin tutkielmassa on esitetty trakealima- ja keuhkohuutelunäytteiden kerääminen, laboratoriotutkimukset ja tulosten tulkintaa. Lisäksi kirjallisuuskatsauksessa on vertailtu näitä kahta menetelmää. Tutkimusosuuden tarkoituksena oli pyrkiä selvittämään, kumpi näytteenottomenetelmä olisi diagnostisempi yliopistollisen hevossairaalan potilasmateriaalissa. Potilastiedostosta etsittiin 51 yliopistolliseen hevossairaalaan hengitystiesairauteen viittaavan oireilun takia tutkittavaksi tullutta hevosta, joilta oli otettu sekä trakealima- että keuhkohuuhtelunäyte. Hevosilta oli arvioitu hengitysteiden liman määrä. Näytteiden solukuva tutkittiin yliopistollisen eläinsairaalan keskuslaboratoriossa. Hevosilta oli määritetty lisäksi veren fibrinogeenipitoisuus ja valkosolujen määrä. Hevoset olivat iältään 2-25 vuotiaita, ja mukana oli tammoja, ruunia ja oreja. Rodultaan hevoset olivat puoliverisiä, lämminverisiä, suomenhevosia ja poneja. Yleisin oire oli alentunut suorituskyky. Muita oireita olivat yskä, räkäisyys, nenäverenvuoto ja kuume. Näytteenottomenetelmien välillä havaittiin riippuvuus neutrofiilien, lymfosyyttien ja makrofagien prosenttiosuuksissa. Hengitysteiden liman määrän ja neutrofiilien prosenttiosuuksien välillä havaittiin riippuvuus. Fibrinogeenipitoisuuden ja liman määrän välillä, sekä fibrinogeenin ja trakealimanäytteiden neutrofiilien välillä havaittiin myös riippuvuus. 30/51 määriteltiin sairaiksi jomman kumman näytteen perusteella käytetyillä neutrofiilien raja-arvoilla, mutta 9/51 hevosella (18 %) trakealima- ja keuhkohuuhtelunäytteen tulkinta erosi toisistaan. Tulokset viittaavat siihen, että tässä potilasaineistossa trakealima- ja keuhkohuuhtelunäytteet olisivat sairauden havaitsemisen suhteen melko vertailukelpoisia keskenään tämänhetkistä näytteiden tulkintatapaa käytettäessä. Tutkimuksen aineisto oli hyvin valikoitunutta; kaikilla hevosilla oireilu oli ollut kroonista tai uusiutuvaa. Tutkimuksessa ei ollut mukana oireetonta vertailuryhmää. Tässä tutkimuksessa menetelmien herkkyyttä ja tarkkuutta sairauden havaitsemisen suhteen ei voitu määrittää, sillä sairautta ei pystytty toteamaan ilman, että näytteiden tulokset olisivat olleet osana diagnoosia. Lisää tutkimuksia näytteenottomenetelmien osuvuuden määrittämiseksi tarvitaan

Comparison of Tracheal Wash and Bronchoalveolar Lavage Cytology in 154 Horses With and Without Respiratory Signs in a Referral Hospital Over 2009−2015

Most equine lower respiratory diseases present as increased airway neutrophilia, which can be detected in tracheal wash (TW) or bronchoalveolar lavage fluid (BALE) cytology samples. The aim was to compare the TW and BALF results in a population of client-owned horses with and without clinical respiratory disease signs. A secondary aim was to determine the sensitivity (Se) and specificity (Sp) of TW and BALF neutrophilia in detecting respiratory disease. The cutoff values for neutrophils were also evaluated. Retrospective data from 154 horses of various breeds that had been subject to both TW and bronchoalveolar lavage (BAL) sampling at rest during 2009-2015 were used. The horses were divided into three groups based on the presenting signs, physical examination, and endoscopy mucus score. Neutrophil counts of >20% in TW and >5% in BAL were considered abnormal. Cytology results between groups, correlations between 1W and BALF cell types, and tracheal mucus score were analyzed. Two graph receiving operating characteristic (ROC) curves of the neutrophil percentage values of TW and BALF were created to determine the optimal cutoff values and to calculate the diagnostic Se and Sp for diagnosing airway inflammation in horses with and without clinical respiratory signs. The Se and Sp of TW and BALF neutrophil percentages were further estimated using a two-test one-population Bayesian latent class model. The two tests showed substantial agreement, and only 17.5% of the horses were classified differently (healthy vs. diseased). The neutrophil percentage was found to correlate between TW and BALF. The Se and Sp of TW were generally higher than for BAL when estimated with area under the curve or Bayesian model. Cutoff values of 17.7% for TW and 7% for BALF were indicated by the ROCs. We conclude that TW is a more sensitive and specific method in our patient population. We suggest that the current neutrophil cutoff values of 20% for 1W and 5% for BALE would still be appropriate to use in clinical diagnosis of airway inflammation. However, further studies with other cell types and in other populations are warranted to determine the best sampling method for individual horses.Peer reviewe

Biologiset torjuntavalmisteet perunaseitin torjunnassa

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Genome-wide association studies highlight novel risk loci for septal defects and left-sided congenital heart defects

Abstract Background Congenital heart defects (CHD) are structural defects of the heart affecting approximately 1% of newborns. They exhibit low penetrance and non-Mendelian patterns of inheritance as varied and complex traits. While genetic factors are known to play an important role in the development of CHD, the specific genetics remain unknown for the majority of patients. To elucidate the underlying genetic risk, we performed a genome wide association study (GWAS) of CHDs in general and specific CHD subgroups using the FinnGen Release 10 (R10) (N > 393,000), followed by functional fine-mapping through eQTL and co-localization analyses using the GTEx database. Results We discovered three genome-wide significant loci associated with general CHD. Two of them were located in chromosome 17: 17q21.32 (rs2316327, intronic: LRRC37A2, Odds ratio (OR) [95% Confidence Interval (CI)] = 1.17[1.12–1.23], p = 1.5 × 10–9) and 17q25.3 (rs1293973611, nearest: BAHCC1, OR[95%CI] = 4.48[2.80–7.17], p = 7.0 × 10–10), respectively, and in addition to general CHD, the rs1293973611 locus was associated with the septal defect subtype. The third locus was in band 1p21.2 (rs35046143, nearest: PALMD, OR[95%CI] = 1.15[1.09–1.21], p = 7.1 × 10–9), and it was associated with general CHD and left-sided lesions. In the subgroup analysis, two additional loci were associated with septal defects (rs75230966 and rs6824295), and one with left-sided lesions (rs1305393195). In the eQTL analysis the variants rs2316327 (general CHD), and rs75230966 (septal defects) both located in 17q21.32 (with a LD r2 of 0.41) were both predicted to significantly associate with the expression of WNT9B in the atrial appendage tissue category. This effect was further confirmed by co-localization analysis, which also implicated WNT3 expression in the atrial appendage. A meta-analysis of general CHD together with the UK Biobank (combined N = 881,678) provided a different genome-wide significant locus in LRRC37A2; rs16941382 (OR[95%CI] = 1.15[1.11–1.20], p = 1.5 × 10–9) which is in significant LD with rs2316327. Conclusions Our results of general CHD and different CHD subcategories identified a complex risk locus on chromosome 17 near BAHCC1 and LRRC37A2, interacting with the genes WNT9B, WNT3 and MYL4, may constitute potential novel CHD risk associated loci, warranting future experimental tests to determine their role

Genome-wide association studies highlight novel risk loci for septal defects and left-sided congenital heart defects

Background Congenital heart defects (CHD) are structural defects of the heart affecting approximately 1% of newborns. They exhibit low penetrance and non-Mendelian patterns of inheritance as varied and complex traits. While genetic factors are known to play an important role in the development of CHD, the specific genetics remain unknown for the majority of patients. To elucidate the underlying genetic risk, we performed a genome wide association study (GWAS) of CHDs in general and specific CHD subgroups using the FinnGen Release 10 (R10) (N > 393,000), followed by functional fine-mapping through eQTL and co-localization analyses using the GTEx database. Results We discovered three genome-wide significant loci associated with general CHD. Two of them were located in chromosome 17: 17q21.32 (rs2316327, intronic: LRRC37A2, Odds ratio (OR) [95% Confidence Interval (CI)] = 1.17[1.12–1.23], p = 1.5 × 10–9) and 17q25.3 (rs1293973611, nearest: BAHCC1, OR[95%CI] = 4.48[2.80–7.17], p = 7.0 × 10–10), respectively, and in addition to general CHD, the rs1293973611 locus was associated with the septal defect subtype. The third locus was in band 1p21.2 (rs35046143, nearest: PALMD, OR[95%CI] = 1.15[1.09–1.21], p = 7.1 × 10–9), and it was associated with general CHD and left-sided lesions. In the subgroup analysis, two additional loci were associated with septal defects (rs75230966 and rs6824295), and one with left-sided lesions (rs1305393195). In the eQTL analysis the variants rs2316327 (general CHD), and rs75230966 (septal defects) both located in 17q21.32 (with a LD r2 of 0.41) were both predicted to significantly associate with the expression of WNT9B in the atrial appendage tissue category. This effect was further confirmed by co-localization analysis, which also implicated WNT3 expression in the atrial appendage. A meta-analysis of general CHD together with the UK Biobank (combined N = 881,678) provided a different genome-wide significant locus in LRRC37A2; rs16941382 (OR[95%CI] = 1.15[1.11–1.20], p = 1.5 × 10–9) which is in significant LD with rs2316327. Conclusions Our results of general CHD and different CHD subcategories identified a complex risk locus on chromosome 17 near BAHCC1 and LRRC37A2, interacting with the genes WNT9B, WNT3 and MYL4, may constitute potential novel CHD risk associated loci, warranting future experimental tests to determine their role.peerReviewe

Evidence for three genetic loci involved in both anorexia nervosa risk and variation of body mass index

The maintenance of normal body weight is disrupted in patients with anorexia nervosa (AN) for prolonged periods of time. Prior to the onset of AN, premorbid body mass index (BMI) spans the entire range from underweight to obese. After recovery, patients have reduced rates of overweight and obesity. As such, loci involved in body weight regulation may also be relevant for AN and vice versa. Our primary analysis comprised a cross-trait analysis of the 1000 single-nucleotide polymorphisms (SNPs) with the lowest P-values in a genome-wide association meta-analysis (GWAMA) of AN (GCAN) for evidence of association in the largest published GWAMA for BMI (GIANT). Subsequently we performed sex-stratified analyses for these 1000 SNPs. Functional ex vivo studies on four genes ensued. Lastly, a look-up of GWAMA-derived BMI-related loci was performed in the AN GWAMA. We detected significant associations (P-values <5 × 10-5, Bonferroni-corrected P<0.05) for nine SNP alleles at three independent loci. Interestingly, all AN susceptibility alleles were consistently associated with increased BMI. None of the genes (chr. 10: CTBP2, chr. 19: CCNE1, chr. 2: CARF and NBEAL1; the latter is a region with high linkage disequilibrium) nearest to these SNPs has previously been associated with AN or obesity. Sex-stratified analyses revealed that the strongest BMI signal originated predominantly from females (chr. 10 rs1561589; Poverall: 2.47 × 10-06/Pfemales: 3.45 × 10-07/Pmales: 0.043). Functional ex vivo studies in mice revealed reduced hypothalamic expression of Ctbp2 and Nbeal1 after fasting. Hypothalamic expression of Ctbp2 was increased in diet-induced obese (DIO) mice as compared with age-matched lean controls. We observed no evidence for associations for the look-up of BMI-related loci in the AN GWAMA. A cross-trait analysis of AN and BMI loci revealed variants at three chromosomal loci with potential joint impact. The chromosome 10 locus is particularly promising given that the association with obesity was primarily driven by females. In addition, the detected altered hypothalamic expression patterns of Ctbp2 and Nbeal1 as a result of fasting and DIO implicate these genes in weight regulation