Comparative evaluation of trypsin and elastase digestion techniques for isolation of murine retinal vasculature.

Dysfunctional pericytes and disruption of adherens or tight junctions are related to many microvascular diseases, including diabetic retinopathy. In this context, visualizing retinal vascular architecture becomes essential for understanding retinal vascular disease pathophysiology. Although flat mounts provide a demonstration of the retinal blood vasculature, they often lack a clear view of microaneurysms and capillary architecture. Trypsin and elastase digestion are the two techniques for isolating retinal vasculatures in rats, mice, and other animal models. Our observations in the present study reveal that trypsin digestion impacts the association between pericytes and endothelial cells. In contrast, elastase digestion effectively preserves these features in the blood vessels. Furthermore, trypsin digestion disrupts endothelial adherens and tight junctions that elastase digestion does not. Therefore, elastase digestion emerges as a superior technique for isolating retinal vessels, which can be utilized to collect reliable and consistent data to comprehend the pathophysiology of disorders involving microvascular structures.

Udder health in relation to udder and teat morphometry in Holstein Friesian × Sahiwal crossbred dairy cows.

The present investigation involved 261 Holstein Friesian (HF) × Sahiwal cows to study the udder health as well as the udder and teat morphometry. The udder health was defined on the basis of bacteriology and California mastitis test of quarter foremilk. The morphometry parameters included udder fore depth (UFD, distance from the point where the fore udder merges within the abdomen to a point in front of the fore teats at the level of the udder base), udder rear depth (URD, distance from the bottom of the vulva to the base of the rear udder), udder depth (UD, level of the udder base with respect to hock joint), height of the udder from the floor, teat length, teat diameter (at mid of the teat barrel), teat-tip to floor distance and distance between the teats. The URD, UD and height of udder from the floor showed a significant (P < 0.05) relation to the udder health with mastitic cows having larger URD, smaller udder distance from the floor and the udder base placed below the hock joint. The cows with teat length larger than 4.5 cm and teat diameter 3.0 cm or larger had significantly more quarters affected with mastitis. Further, cows with teats placed nearer to the floor were more susceptible to mastitis (P < 0.01). No significant differences were found between healthy and mastitic udders with respect to UFD and distance between teats. It can be concluded that selection for optimum udder and teat morphometry in breeding programmes may help to reduce susceptibility to intramammary infections in HF × Sahiwal cows.