Validity and reproducibility of retinal arteriole and venule diameter measurements: ELSA-Brasil study. A cross-sectional study.

BACKGROUND: Investigation of alterations to retinal microvasculature may contribute towards understanding the role of such changes in the pathophysiology of several chronic non-communicable diseases. The objective here was to evaluate the validity and reproducibility of retinal arteriole and venule diameter measurements made by Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) graders. DESIGN AND SETTING: Cross-sectional study at six teaching and research institutions. METHODS: To evaluate validity, each of 25 retinal images from the University of Wisconsin (gold standard) was measured by five ELSA-Brasil graders. To evaluate reproducibility, 105 images across the spectrum of vessel diameters were selected from 12,257 retinal images that had been obtained between 2010 and 2012, and each image was reexamined by the same grader and by an independent grader. All measurements were made using the Interactive Vessel Analysis (IVAN) software. Bland-Altman plots, paired t tests and intraclass correlation coefficients (ICCs) were analyzed. RESULTS: Mean differences between ELSA-Brasil and gold-standard readings were 0.16 µm (95% CI -0.17-0.50; P = 0.31) for central retinal artery equivalent (CRAE), -0.21 µm (95% CI -0.56-0.14; P = 0.22) for central retinal vein equivalent (CRVE) and 0.0005 (95% CI -0.008-0.009; P = 0.55) for arteriole/venule ratio (AVR). Intragrader ICCs were 0.77 (95% CI 0.67-0.86) for CRAE, 0.90 (95% CI 0.780.96) for CRVE and 0.70 (0.55-0.83) for AVR. Intergrader ICCs were 0.75 (95% CI 0.64-0.85) for CRAE, 0.90 (95% CI 0.79-0.96) for CRVE and 0.68 (95% CI 0.55-0.82) for AVR. CONCLUSIONS: Retinal microvascular diameter measurements are valid and present moderate to high intra and intergrader reproducibility in ELSA-Brasil.

Validity and reproducibility of retinal arteriole and venule diameter measurements: ELSA-Brasil study. A cross-sectional study

ABSTRACT BACKGROUND: Investigation of alterations to retinal microvasculature may contribute towards understanding the role of such changes in the pathophysiology of several chronic non-communicable diseases. The objective here was to evaluate the validity and reproducibility of retinal arteriole and venule diameter measurements made by Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) graders. DESIGN AND SETTING: Cross-sectional study at six teaching and research institutions. METHODS: To evaluate validity, each of 25 retinal images from the University of Wisconsin (gold standard) was measured by five ELSA-Brasil graders. To evaluate reproducibility, 105 images across the spectrum of vessel diameters were selected from 12,257 retinal images that had been obtained between 2010 and 2012, and each image was reexamined by the same grader and by an independent grader. All measurements were made using the Interactive Vessel Analysis (IVAN) software. Bland-Altman plots, paired t tests and intraclass correlation coefficients (ICCs) were analyzed. RESULTS: Mean differences between ELSA-Brasil and gold-standard readings were 0.16 µm (95% CI -0.17-0.50; P = 0.31) for central retinal artery equivalent (CRAE), -0.21 µm (95% CI -0.56-0.14; P = 0.22) for central retinal vein equivalent (CRVE) and 0.0005 (95% CI -0.008-0.009; P = 0.55) for arteriole/venule ratio (AVR). Intragrader ICCs were 0.77 (95% CI 0.67-0.86) for CRAE, 0.90 (95% CI 0.780.96) for CRVE and 0.70 (0.55-0.83) for AVR. Intergrader ICCs were 0.75 (95% CI 0.64-0.85) for CRAE, 0.90 (95% CI 0.79-0.96) for CRVE and 0.68 (95% CI 0.55-0.82) for AVR. CONCLUSIONS: Retinal microvascular diameter measurements are valid and present moderate to high intra and intergrader reproducibility in ELSA-Brasil.

The histological analysis of the glenohumeral "critical zone" / Análisis histológico de la "zona crítica" glenohumeral

The arterial integrity of the "critical zone" of the rotator cuff has led to much uncertainty regarding rotator cuff tendinopathy. As the region of the supraspinatus tendon is the most common area affected by impingement, its central aspect is situated approximately 10 mm from the insertion at the greater humeral tubercle. Although many studies have investigated the vascularity of the "critical zone", there still appears to be lack of consensus regarding its extent. Through the employment of gross dissection and standard histology analysis of twenty-five adult bilateral cadaveric scapulo-humeral regions (n = 50), this study aimed to quantify the degree of vascularity, or lack thereof, within the "critical zone" by evaluating its relative morphometric features. The demographic representation of the sample was also considered. Results: i) Mean diameter of arteriole lumen: 91.6±75.2 µm; ii) Mean diameter of entire arteriole: 119.8±87.1 µm; iii) Mean arteriole wall thickness: 15.1±9.5 µm; iv) Mean area occupied by an arteriole: 20644.4±3358.0 µm2; v) Mean number of arterioles within "critical zone": 14.6±8.7. All tissue samples displayed a scarce distribution of arterioles along the musculo-tendinous junction relative to the "critical zone". A directly proportional relationship between the morphometric parameters was indicated by positive strong correlations and accompanying statistically significant P values. As 66 % of the number of arterioles within the "critical zone" were distributed between the minimum value and the upper quartile, it was postulated to be hypovascular, therefore confirming the findings of previous studies.

La integridad arterial de la "zona crítica" del manguito rotador ha dado lugar a una gran incertidumbre con respecto a la tendinitis del mismo. A medida que la región del tendón del músculo supraespinoso sea el área más común afectada por el impacto, su aspecto central estará situado aproximadamente a 10 mm desde la inserción en el tubérculo mayor del húmero. Aunque muchos estudios han investigado la vascularización de la "zona crítica", aún parece existir falta de consenso en cuanto a su extensión. A través de la disección macroscópica y análisis de la histología normal de ambas regiones escápulo-humerales, se realizó en 25 cadáveres adultos (n = 50), este estudio que tuvo como objetivo cuantificar el grado de vascularización, o su ausencia, dentro de la "zona crítica" mediante la evaluación de su características morfométricas. También se consideró la representación demográfica de la muestra. Resultados: i) La media de diámetro de lumen de las arteriolas: 91,6 ± 75.2 µm; ii) El diámetro medio de la totalidad de las arteriolas: 119,8 ± 87.1 µm; iii) la media del espesor de la pared de las arteriolas: 15,1 ± 9.5µm; iv) La media de la zona ocupada por una arteriola: 20.644,4 ± 3358.0 µm2; v) La media de las arteriolas dentro de la "zona crítica": 14,6 ± 8,7. Todas las muestras de tejidos tenían una escasa distribución de las arteriolas a lo largo de la unión músculo-tendinosa con relación a la "zona crítica". Una relación directamente proporcional entre los parámetros morfométricos indica fuertes correlaciones positivas y junto a valores de P estadísticamente significativos. Como el 66 % de las arteriolas dentro de la "zona crítica" se distribuyeron entre el valor de cuartil mínimo y superior, se postuló como hipovascular, por lo tanto confirma los resultados de estudios anteriores.

Effect of gender on training-induced vascular remodeling in SHR

There is accumulating evidence that physical inactivity, associated with the modern sedentary lifestyle, is a major determinant of hypertension. It represents the most important modifiable risk factor for cardiovascular diseases, which are the leading cause of morbidity and mortality for both men and women. In addition to involving sympathetic overactivity that alters hemodynamic parameters, hypertension is accompanied by several abnormalities in the skeletal muscle circulation including vessel rarefaction and increased arteriole wall-to-lumen ratio, which contribute to increased total peripheral resistance. Low-intensity aerobic training is a promising tool for the prevention, treatment and control of high blood pressure, but its efficacy may differ between men and women and between male and female animals. This review focuses on peripheral training-induced adaptations that contribute to a blood pressure-lowering effect, with special attention to differential responses in male and female spontaneously hypertensive rats (SHR). Heart, diaphragm and skeletal muscle arterioles (but not kidney arterioles) undergo eutrophic outward remodeling in trained male SHR, which contributed to a reduction of peripheral resistance and to a pressure fall. In contrast, trained female SHR showed no change in arteriole wall-to-lumen ratio and no pressure fall. On the other hand, training-induced adaptive changes in capillaries and venules (increased density) were similar in male and female SHR, supporting a similar hyperemic response to exercise.

Effect of gender on training-induced vascular remodeling in SHR.

There is accumulating evidence that physical inactivity, associated with the modern sedentary lifestyle, is a major determinant of hypertension. It represents the most important modifiable risk factor for cardiovascular diseases, which are the leading cause of morbidity and mortality for both men and women. In addition to involving sympathetic overactivity that alters hemodynamic parameters, hypertension is accompanied by several abnormalities in the skeletal muscle circulation including vessel rarefaction and increased arteriole wall-to-lumen ratio, which contribute to increased total peripheral resistance. Low-intensity aerobic training is a promising tool for the prevention, treatment and control of high blood pressure, but its efficacy may differ between men and women and between male and female animals. This review focuses on peripheral training-induced adaptations that contribute to a blood pressure-lowering effect, with special attention to differential responses in male and female spontaneously hypertensive rats (SHR). Heart, diaphragm and skeletal muscle arterioles (but not kidney arterioles) undergo eutrophic outward remodeling in trained male SHR, which contributed to a reduction of peripheral resistance and to a pressure fall. In contrast, trained female SHR showed no change in arteriole wall-to-lumen ratio and no pressure fall. On the other hand, training-induced adaptive changes in capillaries and venules (increased density) were similar in male and female SHR, supporting a similar hyperemic response to exercise.

ETA receptor blockade decreases vascular superoxide generation in DOCA-salt hypertension.

Development and progression of end-organ damage in hypertension have been associated with increased oxidative stress. Superoxide anion accumulation has been reported in deoxycorticosterone acetate (DOCA)-salt hypertension, in which endothelin-1 plays an important role in cardiovascular damage. We hypothesized that blockade of ETA receptors in DOCA-salt rats would decrease oxidative stress. Both systolic blood pressure (SBP, 210+/-9 mm Hg; P<0.05) and vascular superoxide generation in vivo were increased in DOCA-salt (44.9+/-10.3% of ethidium bromide-positive nuclei; P<0.05) versus control uninephrectomized (UniNx) rats (118+/-3 mm Hg; 18.5+/-3%, respectively). In DOCA-salt rats, the ETA antagonist BMS 182874 (40 mg/kg per day PO) lowered SBP (170+/-4 versus UniNx, 120+/-3 mm Hg) and normalized superoxide production (21.7+/-6 versus UniNx, 11.9+/-7%). Vitamin E (200 mg/kg per day PO) decreased superoxide formation in DOCA-salt rats (18.8+/-7%) but did not alter SBP. Oxidative stress in nonstimulated circulating polymorphonuclear cells (PMNs) or in PMNs treated with zymosan, an inducer of superoxide release, was similar in DOCA-salt and UniNx groups. Superoxide formation by PMNs was unaffected by treatment with BMS 182874. Western blot analysis showed increased nitrotyrosine-containing proteins in mesenteric vessels from DOCA-salt compared with UniNX. Treatment with either BMS 182874 or vitamin E abolished the differences in vascular nitrotyrosine-containing proteins between DOCA-salt and UniNX. Maximal relaxation to acetylcholine was decreased in DOCA-salt aortas (75.8+/-4.2% versus UniNx, 95.4+/-1.9%, P<0.05). BMS 182874 treatment increased acetylcholine-induced relaxation in DOCA-salt aortas to 93.5+/-4.5%. These in vivo findings indicate that increased vascular superoxide production is associated with activation of the endothelin system through ETA receptors in DOCA-salt hypertension, in apparently blood pressure-independent fashion.

Melatonin and N-acetylserotonin inhibit leukocyte rolling and adhesion to rat microcirculation.

The hormone melatonin produced by the pineal gland during the daily dark phase regulates a variety of biological processes in mammals. The aim of this study was to determine the effect of melatonin and its precursor N-acetylserotonin on the microcirculation during acute inflammation. Arteriolar diameter, blood flow rate, leukocyte rolling and adhesion were measured in the rat microcirculation in situ by intravital microscopy. Melatonin alone or together with noradrenaline did not affect the arteriolar diameter or blood flow rate. Melatonin inhibited both leukocyte rolling and leukotriene B(4) induced adhesion while its precursor N-acetylserotonin inhibits only leukocyte adhesion. The rank order of potency of agonists and antagonist receptor selective ligands suggested that the activation of MT(2) and MT(3) melatonin binding sites receptors modulate leukocyte rolling and adhesion, respectively. The effect of melatonin and N-acetylserotonin herein described were observed with concentrations in the range of the nocturnal surge, providing the first evidence for a possible physiological role of these hormones in acute inflammation.

Effects of Ringer-acetate and Ringer-dextran solutions on the microcirculation after LPS challenge: observations in the hamster cheek pouch.

The effects of NaCl 0.9%, Ringer-acetate, and Ringer-dextran given as intravenous infusions in the microcirculatory changes observed in early stages of endotoxemia were investigated in male hamsters treated with Escherichia coli lipopolysaccharide (LPS). The cheek pouch was studied in vivo by means of intravital microscopy. Mean arterial (MAP) and venous pressures (CVP), heart rate, mean arteriolar internal diameter, spontaneous arteriolar vasomotion (AV), red blood cell velocity (RBCV) in these vessels, and long-term effects of LPS were evaluated in animals treated with either LPS alone or the combination of LPS with NaCl 0.9%, Ringer-acetate and Ringer-dextran. The intravenous injection of LPS (0.3 mg/kg) elicited a significant reduction in MAP and CVP, cessation of AV and a decrease in RBCV. In our study, the heart rate and the arteriolar diameter did not change significantly, compared with the control values obtained before the LPS injection. No improvement in the MAP could be detected with infusions of NaCl 0.9% or Ringer-acetate but the infusion of Ringer-dextran increased it significantly. All infusions tested maintained the CVP until the end of the observation period and the Ringer-dextran increased it significantly. The heart rate was maintained around 360 beats/min with a tendency to decrease 70 min after the LPS infusion in all groups studied except the group which received NaCl 0.9% where the heart rate decreased significantly. In all the four groups, the mean arteriolar diameter did not change significantly with time during the observed period. RBCV decrease with the combination LPS + NaCl 0.9% and the infusions of Ringer-acetate and Ringer-dextran maintained it until the end of the observation period. The combination of LPS + NaCl 0.9% maintained the spontaneous arteriolar vasomotion during 50 min after LPS injection and the infusion of Ringer-acetate maintained it for the 3-h observation period. The infusion of Ringer-dextran maintained the amplitude of the spontaneous arteriolar vasomotion and increased its frequency significantly. The long-term effects of LPS showed weight loss and pus on the periorbital area. Our results suggest that the best solution to maintain the microcirculatory parameters during the early stage of endotoxemia after LPS injection was the Ringer-dextran.

Morphometric analysis of the anastomosing arteriolar network in cat sartorius muscle.

Topological and geometrical characteristics of the anastomotic arteriolar network in cat sartorius muscle were studied. The vessels were dilated and filled with gelatin-ink solution and the muscle cleared with methylsalicylate. The analysis was done on 11 muscles and included 2297 vascular segments and 772 vascular loops classified according to their position within the network. On average, each muscle had 644 transverse arterioles arising from the anastomotic vessels. The length of vascular segments close to feeding arteries was greater than those located in the central region, while the number of transverse arterioles per unit length of arcade vessel showed an opposite tendency. Most vessel orders had similar diameters, except for the segments at the periphery of the muscle, which were significantly larger. No correlation was found between vessel length and diameter. Vascular loops located in the central part of the network were smaller, as assessed by area, perimeter, number of segments, segment length and number of branches. The variability of the parameters between muscles was smaller than variability within each muscle. We concluded that, in addition to the parameters previously reported, quantitative descriptions of anastomotic networks may be enhanced by considering certain topological aspects of microvessels. The separation of segments and loops according to the position in the network may reveal differences between muscles of different sizes and functions which would not be detected if the vessels were considered as a single group.