Blood lactate levels in 31 female dogs with pyometra

<p>Abstract</p> <p>Background</p> <p>Canine pyometra is a life-threatening disease common in countries where spaying of dogs is not routinely performed. The disease is associated with endotoxemia, sepsis, systemic inflammatory response syndrome (SIRS) and a 3–4% mortality rate. Blood lactate analysis is clinically valuable in predicting prognosis and survival, evaluating tissue perfusion and treatment response in human and veterinary critical care settings. The aims of the present study were to investigate 1) the blood lactate levels of female dogs with pyometra by a hand-held analyser and 2) if these levels are related with the clinical status or other biochemical or hematological disorders.</p> <p>Methods</p> <p>In total 31 female dogs with pyometra admitted for surgical ovariohysterectomy and 16 healthy female control dogs were included in the present study. A complete physical examination including SIRS-status determination was performed. Blood samples for lactate concentrations, hematological and biochemical parameters, acid-base and blood gas analysis and other laboratory parameters were collected and subsequently analysed. The diagnosis pyometra was verified with histopathological examination of the uterus and ovaries. Increased hospitalisation length and presence of SIRS were used as indicators of outcome.</p> <p>Results</p> <p>In the pyometra group the median blood lactate level was 1,6 mmol l^-1(range <0.8–2.7 mmol l^-1). In the control group the median lactate level was 1,2 mmol l^-1(range <0.8–2.1 mmol l^-1). Of the 31 bitches 19 (61%) fulfilled 2 or more criteria for SIRS at inclusion, 10 bitches (32%) fulfilled 3 of the SIRS criteria whereas none accomplished more than 3 criteria. Lactate levels did not differ significantly between the pyometra and control group, or between the SIRS positive and SIRS negative dogs with pyometra. Increased lactate concentration (>2.5 mmol l^-1) was demonstrated in one female dog with pyometra (3%), and was not associated with longer hospitalisation or presence of SIRS. Lactate measurement was not indicative of peritonitis. None of the bitches died during or within two months of the hospital stay. The measurements of temperature, heart rate, respiratory rate, percentage bandforms of neutrophilic granulocytes, α₂-globulins, creatinin, pvCO₂, TCO₂and base excess showed significant differences between the SIRS positive and the SIRS negative pyometra cases.</p> <p>Conclusion</p> <p>Increased blood lactate concentrations were demonstrated in 3% (1/31), and SIRS was present in 61% (19/31) of the female dogs with pyometra. Preoperative lactate levels were not related with presence of SIRS or prolonged hospitalisation. Lactate measurement was not indicative of peritonitis. The value of a single and repeated lactate analysis in more severely affected cases remains to be determined.</p

Flowchart outlining the pipeline for small RNAseq analysis

1 figureIncluding the identification of known and putative novel miRNAs, miRNA abundance profiling and differential abundance analysis. rRNA: ribosomal RNA; tRNA: transfer RNA; snoRNA: small nucleolar RNA; snRNA: small nuclear RNA; RE: repeat elements; qPCR: quantitative real-time PCR.Peer reviewe

miRNAs selected for qPCR verification

1 table.The table includes for each miRNA the arguments for its selection for further validation, the forward and reverse sequence, the miRBase sequence used as template for primer design, if successful miRNA amplification was obtained with qPCR and qPCR amplification efficiency.Peer reviewe

Detailed list of known and putative de novo miRNAs that passed the defined quality control filters in the feline urine datasets

1 table.Including the miRNA ID name based on sequence homology with other species (blastn), miRNA coordinates, strand, closest homology miRNA with their percentage of identity and probability (E-value) and consensus mature, star and precursor sequences.Peer reviewe

Principal component analysis (PCA) of samples profiled by small RNAseq technique

A. PCA of urine samples on the basis of normalized read counts of the known and putative novel miRNAs for the 38 samples initially processed. The red arrows indicate the outlier Control samples (C5, C6 and C7). B. PCA excluding the high outlier samples. CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction.Peer reviewe

Detailed characteristics of the known and putative novel miRNAs in cat urine for the 35 samples based on RNAseq data

A. Proportion of samples for which each of the known miRNAs across the different groups were detected. B. Cumulative abundance of the known feline miRNAs. The dots indicate the log10 of the miRNA abundance for each miRNA. miRNAs are sorted in each group in a decreasing order by their miRNA abundance on the x-axis, independently for each group. C. Proportion of samples for which each of the putative novel miRNA candidates across the different groups were detected. D. Cumulative abundance of the putative novel miRNAs. The dots indicate the log10 of the miRNA abundance for each miRNA. miRNAs are sorted in each group in a decreasing order by their miRNA abundance on the x-axis, independently for each group. CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction, CPM: Counts per million.Peer reviewe

Pathway enrichment analysis of putative mRNA target genes of non-redundant differentially abundant (DA) miRNAs seeds surviving the criteria |log2FC| ≥ 1.5 and q-value < 0.05 in qPCR analyses and which were also DA in the small RNAseq dataset

1 table.Pathway enrichment of the KEGG and Reactome terms are shown if they gathered significant false discovery rate (FDR) and corrected p-values (q-value < 0.05); their associated mRNA genes found are also shown.Peer reviewe

Mean abundance and standard deviation of the miRNAs detected in the urinary miRNAome of the final 35 feline samples included in RNAseq analyses

1 table.This includes information from their source (database/study/de novo) and their genomic coordinates. CPM: Counts Per Million.Peer reviewe

Pearson correlation analysis between abundance profiles of small RNAseq and qPCR data from selected miRNAs that were DA (|log2FC| ≥ 1.5 for qPCR and ≥ 2 for small RNAseq; q-value < 0.05) using both methodologies

1 figure.CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction, CPM: Counts per million, Rq: Relative quantities.Peer reviewe