Antimicrobial Activity of Plasma Rich in Platelets (PRP) on the Ocular Microbiota of Healthy Horses from Minas Gerais

In equine ophthalmology, ulcerative keratitis is among the most common conditions and, in general, arises as a consequence of some trauma suffered. Secondarily, subsequent contamination by pathogenic or resident bacteria of the horse\u27s ocular microbiota may have undesirable consequences. Under physiological conditions, the normal microbiota coexists with the immune status of the host, serving as a barrier, ensuring the health of the ocular surface, and inhibiting the proliferation of pathogens. However, in the imbalance of immune barriers, the normal microbiota can become pathogenic and lead to infection, acting as an opportunistic agent. The present study aims to demonstrate the antimicrobial effect of platelet-rich plasma (PRP), its time of action, and its correlation with the concentration of its same components in vitro on Staphylococcus sciuri, a bacterium with high prevalence in the normal ocular microbiota of horses in the municipality of Minas Gerais. For the preparation of the PRP, eight adult Quarter Horse (QH) horses were used. The individual PRP was prepared by the double centrifugation protocol, and then, the PRPs were added to a pool, followed by testing their interaction in culture with Brain Heart Infusion (BHI) broth at different dilutions against five strains collected from different animals. After 3, 6, 12, and 18 hours, the colony formation units (CFU) count on a 5% horse blood agar plate was evaluated for each time point. Our study showed that Staphylococcus sciuri, the resident microorganism of the ocular conjunctival microbiota of horses, is more susceptible when compared to the standard strain “American Type Culture Collection” (ATCC-29213) Staphylococcus aureus, a pathogenic microorganism, which was used for the validation of our study. The antibacterial effect shown in this study was bacteriostatic for up to 6 hours. The most concentrated PRP dilutions, 1 : 1 and 1 : 2, were also most effective, suggesting that the antibacterial effect is volume dependent

Endothelial function and insulin resistance in early postmenopausal women with cardiovascular risk factors: importance of ESR1 and NOS3 polymorphisms.

Cardiovascular benefits from estradiol activation of nitric oxide endothelial production may depend on vascular wall and on estrogen receptor alpha (ESR1) and nitric oxide synthase (NOS3) polymorphisms. We have evaluated the microcirculation in vivo through nailfold videocapillaroscopy, before and after acute nasal estradiol administration at baseline and after increased sheer stress (postocclusive reactive hyperemia response) in 100 postmenopausal women, being 70 controls (healthy) and 30 simultaneously hypertensive and diabetic (HD), correlating their responses to PvuII and XbaI ESR1 polymorphisms and to VNTR, T-786C and G894T NOS3 variants. In HD women, C variant allele of ESR1 Pvull was associated to higher vasodilatation after estradiol (1.72 vs 1.64 mm/s, p = 0.01 compared to TT homozygotes) while G894T and T-786C NOS3 polymorphisms were connected to lower increment after shear stress (15% among wild type and 10% among variant alleles, p = 0.02 and 0.04). The G variant allele of ESR1 XbaI polymorphism was associated to higher HOMA-IR (3.54 vs. 1.64, p = 0.01) in HD and higher glucose levels in healthy women (91.8 vs. 87.1 mg/dl, p = 0.01), in which increased waist and HOMA-IR were also related to the G allele in NOS3 G894T (waist 93.5 vs 88.2 cm, p = 0.02; HOMA-IR 2.89 vs 1.48, p = 0.05). ESR1 Pvull, NOS3 G894T and T-786C polymorphism analysis may be considered in HD postmenopausal women for endothelial response prediction following estrogen therapy but were not discriminatory for endothelial response in healthy women. ESR1 XbaI and G894T NOS3 polymorphisms may be useful in accessing insulin resistance and type 2 diabetes risks in all women, even before menopause and occurrence of metabolic disease

RBCV₂ and RBCV₂ increment in HD women according to <i>ESR1 PvuII</i> and <i>Xbal</i> genotypes.

<p>Data are presented as Whiskers plot. Comparison between groups was made by Mann Whitney test. RBCV₂: Red blood cells velocity (mm/s) after estrogen administration. RBCV₂ increment: Red blood cells velocity increment (%) after 1 min ischemia and subsequent reactive hyperemia response upon occlusion release after estrogen administration. A and C, RBCV₂ and <i>PvuII</i> and <i>Xbal</i> genotypes, respectively. B and D, RBCV₂ increment and <i>PvuII</i> and <i>Xbal</i> genotypes, respectively.</p

RBCV₁ and RBCV₁ increment in HD women according to <i>NOS3 T-786C and G894T</i> genotypes.

<p>Data are presented as Whiskers plot. Comparison between groups was made by Mann Whitney test. RBCV₁: Red blood cells velocity (mm/s). RBCV₁ increment: Red blood cells velocity increment (%) after 1 min ischemia and subsequent reactive hyperemia response upon occlusion release. A, C, and E, RBCV₁ and <i>VNTR</i>, <i>-786C</i>, and <i>G894T</i> genotypes, respectively. B, D, and F, RBCV₁ increment and <i>VNTR</i>, <b><i>-</i></b><i>786C</i>, and <i>G894T</i> genotypes, respectively.</p

Clinical and laboratory data in HD and healthy women.

<p>Reference values: HOMA-IR, 2.1 T 0.7; Triglycerides, <150 mg/dl; HDL cholesterol, >50 mg/dl; hS C-reactive protein, <0.3 mg/dl; Estradiol, <44 pg/ml (post menopause without HT);</p><p>Oxidized LDL, <0.5 nmol/mg ApoPt.</p><p>BMI, body mass index; HOMA-IR, homeostasis model assessment of insulin resistance; RBCV₁: red blood cell velocity before estradiol: RBCV_max1: peak red blood cell velocity before estradiol: RBCV₁ increment: % of RBCV_max1 increase in relation to RBCV₁; TRBCV_max1: time to reach peak red blood cell velocity before estradiol; RBCV₂: red blood cell velocity after estradiol: RBCV_max2: peak red blood cell velocity after estradiol: RBCV₂ increment: % of RBCV_max2 increase in relation to RBCV₂: TRBCV_max2: time to reach peak red blood cell velocity after estradiol. Comparison between groups was performed by Mann Whitney test.</p

Frequencies of <i>ESR1</i> and <i>NOS3</i> polymorphisms in HD and healthy women.

<p>Comparison between genotypes was performed by Fisher’s exact test.</p

Acute Effects of Metformin and Vildagliptin after a Lipid-Rich Meal on Postprandial Microvascular Reactivity in Patients with Type 2 Diabetes and Obesity: A Randomized Trial

Background: Type 2 diabetes mellitus and obesity are both related to endothelial dysfunction. Postprandial lipemia is a cardiovascular risk. Notably, it is known that a high-fat diet may elicit microvascular dysfunction, even in healthy subjects. Since anti-diabetic drugs have different mechanisms of action and also distinct vascular benefits, we aimed to compare the results of two anti-diabetic drugs after the intake of a lipid-rich meal on microcirculation in patients with type 2 diabetes and obesity. In parallel, we also investigated the metabolic profile, oxidative stress, inflammation, plasma viscosity, and some gastrointestinal peptides. Subjects/Methods: We included 38 drug-na&iuml;ve patients, all women aged between 19 and 50 years, with BMI &ge; 30 kg/m2. We performed endothelial measurements and collected samples before (fasting) and after the intake of a lipid-rich meal at 30, 60, 120, and 180 min. Patients were randomized to metformin or vildagliptin, given orally just before the meal. Endothelial function was assessed by videocapillaroscopy and laser-Doppler flowmetry to investigate microvascular reactivity. Besides, we also investigated plasma viscosity, inflammatory and oxidative stress biomarkers, gastrointestinal peptides, and metabolic profile in all time points. Results: No differences at baseline were noted between groups. Vildagliptin increased glucagon-like peptide-1 compared to metformin. Paired comparisons showed that, during the postprandial period, vildagliptin significantly changed levels of insulin and glucagon-like peptide-1, and also the dipeptidyl peptidase-4 activity, while metformin had effects on plasma glucose solely. Metformin use during the test meal promoted an increase in functional capillary density, while vildagliptin kept non-nutritive microvascular blood flow and vasomotion unchanged. Conclusions: After the intake of a lipid-rich meal, the use of vildagliptin preserved postprandial non-nutritive microflow and vasomotion, while metformin increased capillary recruitment, suggesting protective and different mechanisms of action on microcirculation