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Assessment of renal graft function by perioperative monitoring of cortical microcirculation in kidney transplantation.

BACKGROUND: We evaluated the significance of perioperative cortical microperfusion for graft function and long-term prognosis after renal allotransplantation. Thermodiffusion technology was clinically applied for the first time, after previous validation for perfusion monitoring of the renal cortex in pigs. METHODS: A thermodiffusion probe was inserted into the renal cortex in 30 transplant recipients after graft reperfusion. Real-time measurements were recorded until the end of the operation. In 14 patients perfusion was measured daily until postoperative day 7. Microcirculation was correlated to serum creatinine level, scintigraphic findings, and long-term outcome. RESULTS: In primary graft function, intraoperative perfusion was 85+/-7 mL/100 g per min compared with significantly lower values in cases with subsequent graft dysfunction. The best discrimination was defined for a level of 70 mL/100 g per min with a positive predictive value of 88% for detection of good graft function and 86% for nonfunction. Intraoperative perfusion was significantly different in patients with normal grafts, delayed function, and graft loss. Postoperatively, lower perfusion was found in acute tubular necrosis; a significant correlation could be noted between microcirculation and perfusion index measured by nuclear scanning (r=0.78, P<0.01). Living-related grafts were characterized by higher intraoperative perfusion and superior graft quality. CONCLUSION: Thermodiffusion could be clinically applicable for the perioperative monitoring of renal graft perfusion. Intraoperative reduction of cortical microcirculation has a high predictive value with respect to detection of delayed renal function. Postoperatively, impaired renal microperfusion is associated with acute tubular necrosis. Living-related donor grafts show less microcirculatory alteration than cadaveric kidneys.

Adolescent↗

Exudative hyperresponsiveness of the airway microcirculation in seasonal allergic rhinitis.

BACKGROUND: Mucosal exudation of plasma is a non-injurious, physiological response of the airway microcirculation to different inflammatory processes. The exudative response is similar in the nose and bronchi and exudation occurs in both allergic asthma and rhinitis. The exudative response is a specific end-organ function of the mucosal microcirculation that may be altered in airway diseases. OBJECTIVE: This study examines the hypothesis of altered responsiveness of the superficial airway microcirculation to vascular permeability-increasing challenges in sustained allergic inflammation. METHODS: Fourteen patients with birch-pollen induced allergic rhinitis were studied for 7 weeks during a Swedish birch-pollen season. Nasal symptoms (itching, sneezing, blockage, and discharge) were recorded and the occurrence of pollen was determined. The plasma exudation response was examined by topical histamine challenges at the end (May) and well out of (December) the season. Challenge and lavage were carried out concomitantly using a 'nasal pool'-device. The unilateral nasal cavity was filled for consecutive 10 minute periods with saline and two concentrations of histamine (80 micrograms/mL and 400 micrograms/mL). The lavage fluid levels of different-sized plasma proteins (albumin-66,000 D, fibrinogen-340,000 D, and alpha 2-macroglobulin-725,000 D) were determined. RESULTS: The pollen season was mild resulting in only minor nasal symptoms. Histamine produced exudation of all plasma proteins across the microvascular epithelial barriers with particularly strong correlation between the levels of albumin and alpha 2-macroglobulin (r = 0.98; P < 0.001). The exudative response to histamine was concentration-dependent (P < 0.05) and, furthermore, it was significantly greater late into the season compared with outside the pollen season (albumin: P < 0.05, fibrinogen; P < 0.05, alpha 2-macroglobulin: P < 0.01). CONCLUSION: We conclude that histamine produced concentration-dependent nasal airway exudation of bulk plasma in subjects with seasonal rhinitis and that this response is abnormally great during the pollen season. Whether angiogenesis or increased responsiveness of the microvascular endothelium may explain this phenomenon now remains unknown. We suggest that a microvascular exudative hyperresponsiveness may characterize allergic airway disease.

Adult↗

Effect of experimental non-insulin requiring diabetes on myocardial microcirculation during ischaemia in dogs.

To examine whether chronic high blood glucose may influence myocardial microcirculation during acute myocardial ischaemia in the dog, a noninsulin-requiring diabetes was induced by the streptozotocin-alloxan method. Seventy-five days later, myocardial ischaemia was provoked by occluding the left anterior descending coronary artery for 2 h and microcirculation regulation was assessed in the ischaemic and non-ischaemic myocardium by the radioactive microsphere method. Diabetic dogs were compared with normal dogs. Diabetic dogs had higher blood glycated haemoglobin (2.66 +/- 0.4%) and fructosamine (397 +/- 62 mumol l-1) than control dogs (0.66 +/- 0.2, P < 0.004 and 229 +/- 13, P < 0.03, respectively). Haemodynamic data in the two groups were not different at any time. The size of the ischaemic zone was similar in both groups. During the 2 h ischaemia in the ischaemic zone subendocardial (P = 0.22) and subepicardial (P < 0.05) blood flow slightly increased in control dogs whereas there was a 63% (P < 0.02) and 35% (P = 0.06) reduction respectively in diabetic dogs. In the non-ischaemic zone, blood flow of controls tended to increase (P < 0.006 in the subepicardium and P < 0.06 in the subendocardium) whereas in diabetic dogs blood flow tended to decrease (P = 0.03 in the subendocardium and in the subepicardium). This first investigation on myocardial microcirculation in diabetic dogs during ischaemia suggests that one of the possible causes of increased mortality rate from ischaemic cardiac disease in diabetics might be related to a paradoxical and unfavourable pattern of myocardial blood flow during ischaemia.

Animals↗

Tobacco smoke disrupts testicular microcirculation in the rat.

Anaesthetized adult rats were exposed repeatedly to cigarette smoke for 2 s interspersed with exposure to fresh air for either 10, 15 or 30 s using a smoking apparatus. The acute effects of this treatment on testicular microcirculation were studied using laser Doppler flowmetry. Peripheral tissue O2 saturation was measured continuously in the foot during the experiment. Exposure to cigarette smoke caused a moderate acute increase in the testicular blood flow and it inhibited vasomotion. Blood flow and the pattern of vasomotion were normalized after termination of smoke exposure. The time between the last smoke exposure and flow normalization was dependent on the smoke/air dose. Peripheral tissue O2 saturation did not change during the experiments. The effects on testicular microcirculation of local injections of an aqueous extract of cigarette smoke and passive smoking were also studied. Local injection of smoke extract increased blood flow and inhibited vasomotion. Passive smoking caused a moderate decrease in the frequency of vasomotion and amplitude. The observation that cigarette smoke influences testicular microcirculation acutely is discussed in relation to possible adverse effects on spermatogenesis.

Animals↗

In vivo characterization of histamine H1- and H2-receptors in the rat stomach microcirculation.

1 Using a video microscope system, an in vivo microcirculation preparation was designed to characterize histamine receptors in the rat stomach submucosal arterioles (diameter 50 microns). 2 Each in vivo stomach microcirculation preparation received topically applied multiple concentrations of either the selective histamine H1- or H2-receptor agonist and antagonist and the respective responses (changes of the arteriolar diameter) were used to construct the concentration-response curves. 3 Results showed that both 2-thiazolylethylamine and impromidine, the respective selective H1- and H2-receptor agonists, produced concentration-dependent vasodilator responses in the in vivo stomach submucosal arterioles. These vasodilator responses were competitively and selectively blocked by the respective H1-receptor antagonist mepyramine and the H2-receptor antagonist cimetidine. 4 Data from each in vivo preparation were examined separately to yield a Schild plot and a Hill plot, from which the in vivo estimates of the pA2 value, the slope of the Schild plot, and the Hill coefficient were obtained. 5 The estimated pA2 values for mepyramine (9.60 +/- 0.033, mean +/- s.e. mean) and cimetidine (5.98 +/- 0.037) conformed to similar values found in other tissues, showing that the rat stomach microvascular H1- and H2-receptors are of the same nature as similar receptors elsewhere. 6 Both the Hill and the Schild plots yielded regressions with unity slopes, indicating that the agonist-receptor and the antagonist-receptor reactions followed a simple one-to-one stoichiometry. 7 The findings in the present study are discussed and compared with those from other in vitro tissue preparations; it appears that the present in vivo technique is a satisfactory system for characterizing receptors in the vascular smooth muscle of the microcirculation.

Animals↗

Basal cell carcinoma--an in-vivo model of human tumour microcirculation?

The role of angiogenesis in tumour growth and metastasis is well established. However, investigations of tumour microcirculation to date have used either biopsy material from human tumours, or animal models in vivo. We have studied the tumour microcirculation in vivo in human skin cancers using video-microscopy to examine 12 basal cell carcinomas (BCCs) on the head and neck of 11 patients, and compared the vessels with those seen in the peri-lesional skin, and in normal control skin on the opposite side of the body. The vessels seen within the BCCs were markedly abnormal qualitatively, forming bizarre, disorganized patterns. Quantitative analysis revealed that the mean diameter (+/-standard deviation) of the largest vessel was significantly greater within the BCC (0.086+/-0.029 mm) than that in the control skin (0.034+/-0.012 mm) (P<0.001). The area fraction, a measure of the area of tissue occupied by vessels, was increased highly significantly within the BCCs (0.158+/-0.038) compared with both peri-lesional skin (0.029+/-0.012) and control skin (0.027+/-0.010) (P<0.001). Length density, the length of blood vessel per unit area of tissue, was also highly significantly greater within the lesion (210.22+/-66.05 cm(-1)) compared to peri-lesional (27.10+/-15.67 cm(-1)) and control skin (28.27+/-15.81 cm(-1)) (P<0.001). This is the first study to have demonstrated that BCCs possess a distinct tumour microcirculation which can be observed directly, and assessed quantitatively. Prospective studies of tumour progression, possibly after intervention with angiogenesis inhibitors, are possible.

Aged↗

Microcirculation impedance analysis in cat lung.

An analysis of pulsatile microcirculation in cat lung, with special attention to the pulmonary microvascular impedance, is presented. A theoretical calculation is made on the basis of a complete set of experimental data on the morphology and elasticity of cat's pulmonary capillary sheets. The transfer matrix of the pulmonary microvascular impedance is obtained. The input impedance at the capillary entrance and exit are determined. The input impedance at the pulmonary arterial trunk is compared under various physiological conditions. It is shown that although the impact of pulmonary microcirculation on the relationship between the steady mean flow and pressure in the pulmonary arteries and veins is decisively large, the influence of the alveolar microcirculation on the input impedance at the pulmonary arterial trunk is small.

Animals↗

Cutaneous microcirculation in systemic sclerosis and response to intra-arterial reserpine.

The cutaneous microcirculation in the hand was measured in 23 patients with systemic sclerosis, 19 with Raynaud's phenomenon and four without Raynaud's phenomenon, and 20 controls. The patients with Raynaud's phenomenon had a reduced basal blood flow and an exaggerated further reduction on local cold stimulation, though both were normal in patients without Raynaud's phenomenon. Reflex-induced vascular changes by cold stimulation of the contralateral hand showed no differences between the three groups. The blood flows were similar in the affected skin of the anterior chest wall in four patients with systemic sclerosis and peripheral Raynaud's phenomenon and matched controls. In the seven most severely affected patients 1 mg of intra-arterial reserpine produced a prompt improvement in the cutaneous microcirculation which usually lasted one to three weeks but occasionally much longer. By judicious use of repeated injection guided by measurements of the microcirculation it was possible to heal indolent ulcers of the fingers. The results indicate that vascular changes are common in systemic sclerosis but are not fundamental in the pathogenesis of the disease. More probably there is a general soft tissue abnormality that usually but not necessarily affects the vessels.

Cold Temperature↗

ATP-MgCl2 restores renal microcirculation following trauma and severe hemorrhage.

Although ATP-MgCl2 enhances the recovery of renal function after ischemia and reperfusion, it is not known whether this agent has any beneficial effects on renal microcirculation and function in a nonheparinized model of trauma and severe hemorrhage. To study this, a midline laparotomy was performed (i.e., trauma induced) and the rats were bled to and maintained at a mean arterial pressure of 40 mmHg (1 mmHg = 133.32 Pa) until 40% of the maximum shed blood volume was returned in the form of Ringer's lactate (RL) solution. Animals were then resuscitated with 4 times the volume of the shed blood in the form of RL. ATP-MgCl2, 50 mumol/kg body weight, or an equivalent volume of saline, was infused intravenously during and following resuscitation. Renal microcirculation was examined by using colloidal carbon infusion and laser Doppler flow-metry. Glomerular filtration rate (GFR) was assessed with [3H]inulin clearance and cardiac output (CO) was determined by dye dilution technique. The results indicate that the depressed renal microcirculation following hemorrhage and resuscitation was restored by ATP-MgCl2 treatment. GFR was significantly higher in ATP-MgCl2-treated than saline-treated rats. ATP-MgCl2 also increased urine output, restored the decreased CO, and prevented the occurrence of renal edema after hemorrhage and resuscitation. Thus, ATP-MgCl2 appears to be a useful adjunct to crystalloid resuscitation following trauma and severe hemorrhagic shock even in the absence of blood resuscitation.

Adenosine Triphosphate↗

Effects of Choto-san on microcirculation, serum nitric oxide and lipid peroxides in patients with asymptomatic cerebral infarction.

The effects of Choto-san on microcirculation, serum nitric oxide and lipid peroxides were investigated. Fifteen patients with asymptomatic cerebral infarction were investigated before and after a four-week administration of Choto-san. The variables of microcirculation of the bulbar conjunctiva, which were the internal diameter of vessels, flow velocity and flow volume rate, were increased. Serum NO2(-)/NO3(-) tended to increase and lipid peroxides were decreased. Total serum cholesterol was also decreased. These results suggest that Choto-san may improve microcirculation affected by endothelial function and fat metabolism.

Blood Flow Velocity↗

Stimulation and blockade of cholinergic receptors in terminal liver microcirculation in rats.

This study was designed to establish the existence of cholinergic vascular receptors in the terminal portion of the rat liver microcirculation. The liver microcirculation was observed in vivo by a transillumination technique through a television microscope. The changes in the caliber of the liver sinusoids were measured directly on the television screen. Infusion of the parasympathetic neurotransmitter acetylcholine into the portal venous circulation caused a concentration-dependent dilation of liver sinusoids. Similar dilatation effects were observed for other cholinergic receptor agonists. Atropine, the specific cholinergic receptor blocker, inhibited this dilator effect, displacing the acetylcholine concentration-effect curve to the right. In contrast, physostigmine, the cholinesterase inhibitor, caused displacement of the curve to the left. In conclusion, cholinergic receptors are present in the terminal portion of the liver microcirculation, subserving the functional role of vasodilatation.

Acetylcholine↗

Remodeling of the skeletal muscle microcirculation increases resistance to perfusion in obese Zucker rats.

Whereas previous studies have demonstrated that the development of syndrome X in obese Zucker rats (OZR) is associated with impaired arteriolar reactivity to vasoactive stimuli, additional results from these studies indicate that the passive diameter of skeletal muscle arterioles is reduced in OZR versus lean Zucker rats (LZR). On the basis of these prior observations, the present study evaluated structural alterations to the skeletal muscle microcirculation as potential contributors to an elevated vascular resistance. Isolated skeletal muscle resistance arterioles exhibited a reduced passive diameter at all levels of intralumenal pressure and a left-shifted stress-strain curve in OZR versus LZR, indicative of structural remodeling of individual arterioles. Histological analyses using Griffonia simplicifolia I lectin-stained sections of skeletal muscle demonstrated reduced microvessel density (rarefaction) in OZR versus LZR, suggesting remodeling of entire microvascular networks. Finally, under maximally dilated conditions, constant flow-perfused skeletal muscle of OZR exhibited significant elevations in perfusion pressure versus LZR, indicative of an increased resistance to perfusion within the microcirculation. These data suggest that developing structural alterations to the skeletal muscle microcirculation in OZR result in elevated vascular resistance, which may, acting in concert with impaired arteriolar reactivity, contribute to blunted active hyperemic responses and compromised performance of in situ skeletal muscle with elevated metabolic demand.

Algorithms↗

Control of oxidative stress in microcirculation of spontaneously hypertensive rats.

One mechanism for organ damage in individuals with arterial hypertension may be due to oxygen free radical production. This study was designed to localize free radicals in a microvascular network of mature spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto (WKY) rats. Because glucocorticoids play a role in pressure elevation of SHRs, we investigated their role in microvascular free radical formation. Oxygen radical production in mesentery was detected by tetranitroblue tetrazolium reduction to formazan aided by digital light-absorption measurements. Formazan deposits were observed in the endothelial cells and lumens of all microvessels and in lymphatic endothelia but were fewer in tissue parenchyma. The formazan distribution in younger (14-16 wk old) WKY rats and SHRs was heterogeneous with low values in capillaries and small arterioles/venules (<30 microm) but enhanced deposits in larger venules. Adrenalectomy served to reduce the formazan density in SHRs to the level of WKY rats, whereas dexamethasone supplementation of the adrenalectomized rats caused elevation in the larger venules of SHRs. In older (40 wk old) SHRs, formazan levels were elevated in all hierarchies of microvessels. After pressure reduction was employed with chronic hydralazine treatment, the formazan deposits were reduced in all locations of the microcirculation in both WKY rats and SHRs. Elevated formazan deposits were also found in lymphatic endothelium. These results suggest that oxygen free radical production is elevated in both high- and low-pressure regions of SHR microcirculation via a process that is controlled by glucocorticoids. Older SHRs have higher formazan levels than younger SHRs in all microvessels. Chronic hydralazine treatment, which serves to reduce arterial blood pressure, attenuates tetranitroblue tetrazolium reduction in WKY rats and SHRs even in venules of the microcirculation, which has no micropressure elevation. Free radical production may be a more global condition in SHRs and may not be limited to arteries and arterioles.

Adrenal Cortex↗

Mapping of the functional microcirculation in vital organs using contrast-enhanced in vivo video microscopy.

A functional microcirculation is vital to the survival of mammalian tissues. In vivo video microscopy is often used in animal models to assess microvascular function, providing real-time observation of blood flow in normal and diseased tissues. To extend the capabilities of in vivo video microscopy, we have developed a contrast-enhanced system with postprocessing video analysis tools that permit quantitative assessment of microvascular geometry and function in vital organs and tissues. FITC-labeled dextran (250 kDa) was injected intravenously into anesthetized mice to provide intravascular fluorescence contrast with darker red blood cell (RBC) motion. Digitized video images of microcirculation in a variety of internal organs (e.g., lung, liver, ovary, and kidney) were processed using computer-based motion correction to remove background respiratory and cardiac movement. Stabilized videos were analyzed to generate a series of functional images revealing microhemodynamic parameters, such as plasma perfusion, RBC perfusion, and RBC supply rate. Fluorescence contrast revealed characteristic microvascular arrangements within different organs, and images generated from video sequences of liver metastases showed a marked reduction in the proportion of tumor vessels that were functional. Analysis of processed video sequences showed large reductions in vessel volume, length, and branch-point density, with a near doubling in vessel segment length. This study demonstrates that postprocessing of fluorescence contrast video sequences of the microcirculation can provide quantitative images useful for studies in a wide range of model systems.

Animals↗

Stimulated leukocyte adhesion in coronary microcirculation is reduced by a calcium antagonist.

The first step in the acute myocardial inflammatory response is leukocyte sequestration in the coronary microcirculation. To determine the location(s) of stimulated leukocyte deposition in the coronary microcirculation and the effects of the calcium antagonist, nisoldipine, on leukocyte adhesion, leukocytes were stimulated with the chemotactic peptide, N-formylmethionyl-leucyl-phenylalanine (FMLP) and blood cell adherence was evaluated using two methods. In vitro leukostasis was evaluated by measuring the extraction of white cells in nylon fiber columns. We found that diluted whole blood (DWB) demonstrated 30% granulocyte adherence. The chemotactic peptide FMLP (1 microM) significantly increased adherence to 69%. Pretreatment of the blood with nisoldipine (1 microM) immediately before FMLP significantly reduced the FMLP-induced adhesion to 47%. In the coronary microcirculation, FMLP caused a marked increase in leukocyte sequestration, primarily in coronary capillaries. The FMLP effect was somewhat transient because the washout of trapped white cells was similar in the vehicle and FMLP groups. Nisoldipine significantly reduced the FMLP-induced leukostasis in coronary capillaries (P < 0.05). The magnitude of the attenuation of leukostasis with nisoldipine was remarkably similar in both models, suggesting a direct effect of this agent on the blood rather than on the blood vessels. These findings offer another possible mechanism by which dihydropyridine calcium antagonists may be cardioprotective under pathophysiological conditions.

Animals↗

Prolonged leukocyte transit time in coronary microcirculation of endotoxemic pigs.

We quantified the timing and extent of leukocyte retention by the coronary microcirculation in a pig model of hyperdynamic sepsis in three ways. First, the transmyocardial leukocyte gradient was determined as coronary blood flow (calibrated ultrasonic flow probe) multiplied by the difference between leukocyte counts in the aorta and coronary sinus. Measurements were taken at 1-min intervals for 30 min and then at 3-min intervals for 45 min in anesthetized pigs exposed to either endotoxin (50 micrograms/kg iv over 30 min) (n = 7) or vehicle (n = 7). Second, postmortem morphometric analysis was used to quantitate the number and location of retained myocardial leukocytes. Finally, myocardial capillary transit time of leukocytes was calculated from the above measures. In the endotoxin group 2.1 +/- 0.8 x 10(9) leukocytes/100 g wet wt were retained in the coronary circulation, primarily in capillaries. This resulted in 111 +/- 37 (P < 0.05) times as many leukocytes in the coronary microcirculation than predicted from the arterial leukocyte concentration. Myocardial capillary transit time of leukocytes was prolonged to 39.1 +/- 20.6 s (P < 0.05) in the endotoxin group versus 5.0 +/- 1.4 s in the control group. We conclude that, after endotoxin infusion in a pig model of hyperdynamic sepsis, myocardial leukocyte transit is slowed, leading to the retention of large numbers of leukocytes in the coronary microcirculation.

Animals↗

Cytochrome P-450 omega-hydroxylase senses O2 in hamster muscle, but not cheek pouch epithelium, microcirculation.

The goal of this study was to investigate the role of cytochrome P-450 omega-hydroxylase in mediating O2-induced constriction of arterioles in the microcirculation of the hamster. Male Golden hamsters were anesthetized with pentobarbital sodium, and the cremaster muscle or cheek pouch was prepared for observation by intravital microscopy. Arteriolar diameters were measured during elevations of superfusate PO2 from approximately 5 to 150 mmHg. Arteriolar responses to elevated PO2 were determined in the cremaster muscle, in the retractor muscle where it inserts on the cheek pouch, and in the epithelial portion of the cheek pouch. Elevation of superfusion solution PO2 caused a vigorous constriction of arterioles in the cremaster and retractor muscles and in the epithelial portion of the cheek pouch. Superfusion with 10 microM 17-octadecynoic acid, a suicide substrate inhibitor of cytochrome P-450 omega-hydroxylase, and intravenous infusion of N-methylsulfonyl-12,12-dibromododec-11-enamide, a mechanistically different and highly selective inhibitor of cytochrome P-450 omega-hydroxylase, caused a significant reduction in the magnitude of O2-induced constriction of arterioles in the cremaster and retractor muscles. However, arteriolar constriction in response to elevated PO2 was unaffected by 17-octadecynoic acid or N-methylsulfonyl-12,12-dibromododec-11-enamide in the epithelial portion of the cheek pouch. These data confirm that there are regional differences in the mechanism of action of O2 on the microcirculation and indicate that cytochrome P-450 omega-hydroxylase senses O2 in the microcirculation of hamster skeletal muscle, but not in the cheek pouch epithelium.

Amides↗

Expression of adenosine receptors in the preglomerular microcirculation.

The purpose of this study was to systematically investigate the abundance of each of the adenosine receptor subtypes in the preglomerular microcirculation vs. other vascular segments and vs. the renal cortex and medulla. Rat preglomerular microvessels (PGMVs) were isolated by iron oxide loading followed by magnetic separation. For comparison, mesenteric microvessels, segments of the aorta (thoracic, middle abdominal, and lower abdominal), renal cortex, and renal medulla were obtained by dissection. Adenosine receptor protein and mRNA expression were examined by Western blotting, Northern blotting, and RT-PCR. Our results indicate that compared with other vascular segments and renal tissues, A1 and A2B receptor protein and mRNA are abundantly expressed in the preglomerular microcirculation, whereas A2A and A3 receptor protein and mRNA are barely detectable or undetectable in PGMVs. We conclude that, relative to other vascular and renal tissues, A1 and A2B receptors are well expressed in PGMVs, whereas A2A and A3 receptors are notably deficient. Thus A1 and A2B receptors, but not A2A or A3 receptors, may importantly regulate the preglomerular microcirculation.

Animals↗