Search bioRxiv⌕ Search

bioRxiv · 10.1101/633198

Capillary hemoglobin electrophoresis of healthy and anemic dogs: quantification, validation, and reference intervals of hemoglobin fractions

Abstract

Despite the advances in canine medicine and the rapid gaining of attention of canine models in biomedical field and particularly in hemoglobin genes research, the studies on canine hemoglobin composition are sparse with ambiguous findings. Our aim was: i) to investigate the electrophoretic pattern of canine hemoglobin and the possible effect of age, sex, and anemia using a capillary electrophoresis assay, and ii) to validate this assay and calculate reference intervals (RIs) for canine hemoglobin fractions. Blood samples were collected from 53 healthy and 42 dogs with regenerative and non-regenerative anemias. The Sebia Capillarys 2 flex-piercing was used for hemoglobin analysis and it was validated using canine blood samples. R statistical language was employed for the statistical analyses. A major hemoglobin fraction (named HbA0) and a minor one (named HbA2) were identified in 100% and 47.4% of samples, respectively. The within-run and between-run CV was 0.1% for HbA0 and 9.1% and 11.2% for HbA2, respectively. The extremely narrow range of HbA0 and HbA2 values hampered a linearity study using canine blood samples. The RIs for HbA0 and HbA2 were 98.9-100% and 0-1.1%, respectively. HbA0 and HbA2 values were not correlated with age (P=0.866). No differences were observed in the median HbA0 and HbA2 between the two sexes (P=0.823), and healthy and anemic dogs (P=0.805). In conclusion, the capillary electrophoresis revealed a major hemoglobin fraction and an inconsistently present minor fraction. No effect of age, sex, or anemia was detected. The assay used was validated and RIs were generated, so as to be suitable for use in future investigations.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Oikonomidis, I. L., Tsouloufi, T. K., Mylonakis, M. E., Kritsepi-Konstantinou, M.. 2019-05-09. Capillary hemoglobin electrophoresis of healthy and anemic dogs: quantification, validation, and reference intervals of hemoglobin fractions. https://doi.org/10.1101/633198

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Negative association between nodular stromal fibrosis and large lipid droplets in human choroid plexus epithelial cells

Fibrosis is an age-related change within the human choroid plexus that is likely to affect its function negatively. We systematically analyzed nodular stromal fibrosis present in postmortem tissue from a large repository encompassing the human lifespan. Following manual annotation of sampled images from 81 specimens of choroid plexus from the atrium of the lateral ventricle, a deep learning model was trained to assess stages of fibrosis (dense, hyalinized, and mineralized) occurring in papillae across whole slide images of hematoxylin and eosin-stained sections from 204 cases. The percentage of affected papillae and the relative area of fibrosis at all three stages of severity increased with age. Affected papillae were significantly clustered across the choroid plexus, with areas of high and low fibrotic density. Choroid plexus epithelial cells (CPECs) associated with fibrotic papillae were less likely to contain large lipid droplets than were non-fibrotic papillae, as assessed by another deep learning model. Across papillae within a case, the area of each stage of fibrosis was negatively correlated with the prevalence of CPECs containing large lipid droplets. Finally, individuals displaying more nodular fibrosis had significantly fewer CPECs containing large lipid droplets. Thus, age-related nodular stromal fibrosis may negatively impact lipid storage and homeostasis for the choroid plexus and brain.

pathology↗

SilencioM: silencing connected interfering oligonucleotides against MAPT as a potential disease modifying therapeutic for tauopathies

Tauopathies are a diverse group of neurodegenerative diseases whose pathogenesis involves brain accumulation of misfolded, pathological tau (Microtubule Associated Protein Tau, MAPT). More than twenty tauopathies have been identified thus far that collectively impact millions of people worldwide, of which the most common is Alzheimer's disease (AD). Despite substantial efforts to develop drugs for tauopathies, no effective treatments currently exist, highlighting the urgent need for therapeutics to combat these prevalent and ruinous neurodegenerative diseases. Strategies that target tau are likely to have the greatest impact since tau is the proximal cause of their pathogenesis. Small interfering RNAs (siRNAs) have emerged as a powerful class of precision therapeutics that silence disease-causing genes through RNA interference with favorable safety profiles. Delivery challenges are rapidly being addressed, and siRNA-based approaches are poised to transform the therapeutic landscape of neurological diseases. In this study, we describe SilencioM (Silencing connected interfering oligonucleotides against MAPT) and show its efficacy in markedly reducing tau pathology after a single intracerebroventricular injection in PS19 mice, a mouse model of severe tauopathy. SilencioM targets human, primate, rat and mouse ortholog genes; thus, the same compound can proceed from efficacy studies to Investigational New Drug Application (IND)-enabling studies. SilencioM is simple to manufacture and is administered without encapsulation in lipid nanoparticles and without requirements for lipophilic conjugation, making it a promising candidate for further development as a novel therapeutic for tauopathies, including AD.

pathology↗

SilencioMA: A single compound that targets both MAPT and APP as a potential disease modifying therapeutic for Alzheimer's Disease

Alzheimer's Disease (AD) is the most common age-related neurodegenerative disease and the leading cause of dementia worldwide. AD pathogenesis involves aggregation of amyloid-beta (A{beta}), produced from amyloidogenic proteolytic processing of Amyloid Precursor Protein (APP), and accumulation of misfolded, pathological tau (Microtubule Associated Protein Tau, MAPT). Current efforts focus on developing disease modifying therapeutics (DMTs) against single targets: A{beta} or tau. However, DMTs that concurrently target both tau and A{beta} are likely to have the greatest impact since they target both culprits of AD pathogenesis. Here we describe the development of SilencioMA, a single compound that targets both MAPT and APP messenger RNAs (mRNAs) based on silencing connected interfering oligonucleotides against MAPT and APP. Cerebrospinal fluid (CSF) administration of SilencioMA reduces A{beta} and tau and pathology after a single intracerebroventricular injection in 5XFAD/PS19 mice, an animal model of severe amyloidosis and tauopathy. SilencioMA does not rely on lipid nanoparticles or lipophilic conjugates for CSF administration. At the same time its modular design permits conjugation of ligands that facilitate transcytosis through the blood brain barrier, allowing systemic administration. Our findings provide proof-of-concept for a strategy that targets the dual A{beta} and tau pathologies of AD, and establishes a compelling foundation for further development of SilencioMA as a next-generation DMT for AD.

pathology↗