Some properties of androgen-binding activity in rat testis.

High-affinity (Ka approximately equal to 5 X 10(8) M-1 for testosterone) androgen-binding activity in rat testis was shown to have a rapid dissociation rate constant (t1/2 = 3 min, 0 degrees C, 30% glycerol buffer) using dextran-coated charcoal to separate bound from free hormone. Because of this fact, exchange of endogenous and labeled hormone was complete in the assay incubation time (16 h, 0 degrees C) and Scatchard plots of the high-affinity binding data were shown to measure total as contrasted to available sites. The binding was highly specific for androgens. Polyacrylamide gel electrophoresis separated high-affinity androgen-binding protein (Rf 0.54) from albumin (Rf 0.62). Binding site estimates under saturating conditions or by Scatchard analysis of electrophoresis data utilizing [3H]dihydrotestosterone agreed reasonably well with estimates made by the charcoal technique using [3H]testosterone.     

Metabolism of renal microvessels.

Metabolic profiles of renal arteriolar smooth muscle cells were constructed from semiquantitative histochemical analysis of biopsies from 20 mongrel dogs. Vessels ranging in outer wall diameter from 20 to 80 μm were analyzed for the presence and relative intensity of 14 enzymes and 3 substrates with attention being directed in particular to arteriolar smooth muscle cells. Enzymes representing glycolysis, the Krebs cycle, the hexose monophosphate shunt, lipid metabolism, the respiratory chain, and ATP hydrolysis were chosen. Glycogen, hydrophobic lipid, and free fatty acids were also studied. Renal arteriolar cells appear to be chiefly oxidative in their metabolism with a possible preference for free fatty acids as substrate. Anaerobic metabolism seems rather limited in these cells. Finally, arteriolar smooth muscle was found to be quite rich in myosin ATPase.     

Mathematical analysis of hemoglobin spectrophotometry in microvessels.

Spectrophotometry of hemoglobin in microvessels is commonly performed by collecting light either from a small region around the vessel centerline or from the entire lumen of the vessel. In the latter instance, parallel rays of light may not encounter the same amount of absorbing species. Hence, a phenomenon similar to the sieve effect reported in the literature on hemoglobin spectrophotometry may be expected to occur. Although it has been observed that under such circumstances nonlinearities in calibration characteristics arise, the implications of this effect on the interpretation of the spectrophotometric mean concentration have never been addressed so far. Mathematical analysis of hemoglobin spectrophotometry in microvessels, performed in this study, reveals that for practical situations the calibration curve is indeed nonlinear. Moreover, the spectrophotometric mean oxygen saturation is an overestimate of the mean oxygen saturation during oxygenation and an underestimate of the mean oxygen saturation during deoxygenation. These deviations depend upon the manner in which the total heme concentration is distributed within the lumen. Application of the analysis to artificial microvessels showed that the observed superior oxygen transport characteristics of flowing erythrocyte suspensions and hemoglobin solution mixtures could in part be due to the assumptions underlying the procedure used to interpret the experimental results. The implications of this result on models for oxygen transport in microvessels are discussed along with possible resolutions.     

Blood flow velocity in pulmonary microvessels of bullfrog.

Flow velocity in the pulmonary microvessels of the exposed lung of bullfrogs was measured by means of a laser Doppler microscope of an oblique backward mode, together with a signal-analyzing system having a time sharing circuit triggered by the R-wave of the ECG. By these means, measurements of the changes of flow velocity contour in the cardiac cycle were made. Flow velocity was clearly pulsatile in response to cardiac cycles in all microvessels including capillaries. Flow velocities in the arteriole and venule consistently decreased for a short period after the R-wave (84 +/- 33 msec (mean +/- SD) in the arteriole and 130 +/- 31 msec in the venule, respectively) and rapidly increased up to a maximicronm value. The mean flow velocities in arterioles (diameter 50 +/- 17 micron) and venules (39 +/- 9 micronm) were 2.29 +/- 0.32 and 2.30 +/- 0.27 mm/sec. The amplitudes of pulsatile flow in these vessels were 0.83 +/- 0.31 and 0.63 +/- 0.16 mm/sec, respectively. In the capillary the times from the R-wave to the minimicronm and maximum values were variable. In some cases the velocity gradually increased without first decreasing and the increase sharply accelerated a certain time after the R-wave. The mean velocity in the pulmonary capillary and the amplitude of the pulsatile flow ere 1.78 +/- 0.31 and 0.37 +/- 0.12 mm/sec, resepctively. The ratios of the pulsatile amplitude to the mean velocity in the pulmonary capillary, venule and arteriole averaged 0.21, and 0.36, respectively.     

Surface-active phospholipid: a Pandora's box of clinical applications. Part II. Barrier and lubricating properties

In Part I, it was described how their configuration renders phospholipid molecules surface active and capable of acting at interfaces in addition to the liquid-air interface to which conventional theory has hitherto confined the study of 'surfactant' in the lung. Surface-active phospholipid (SAPL) appears no different to comparable surfactants studied in the physical sciences for the highly desirable properties that their adsorption (reversible binding) can impart to solid surfaces. In Part II, these properties are considered in sites where there is no air. Highly desirable properties include boundary lubrication (lubricity), release (antistick) and the ability of the strongly adsorbed and strongly cohesive SAPL linings to act as barriers against abrasion, corrosion and, possibly, against invasion by microorganisms. As the 'sealant', it could be the true barrier rather than the cells providing its mechanical support. Evidence is reviewed for SAPL providing the gastric mucosal barrier to acid in the stomach and preventing the digestion of Helicobacter pylori until that barrier is broken by bile in the duodenum, where H. pylori cause ulcers. The concept that SAPL provides effortless sliding of many tissues, including pleura, pericardium and peritoneum is reviewed. Particular attention is paid to the load-bearing joints, where a deficiency has been associated with osteoarthritis. The ability of the same SAPL lining to perform multiple roles is discussed in relation to the peritoneum, where it could provide the lubricant/release agent preventing surgical adhesions, while imparting semipermeability to 'the membrane' vital for peritoneal dialysis. In each site, the prophylactic use of exogenous SAPL is discussed for its potential clinical applications.     

Angiotensin II receptor binding sites in brain microvessels.

We assessed the specific binding of 125I-labeled angiotensin II (125I-Ang II) to particulate fractions of the cerebral cortex and cerebellum and to microvessels obtained by bulk isolation from these two brain regions in the dog. 125I-Ang II binds to cerebral and cerebellar microvessels in a specific, saturable, and reversible manner and with high affinity (dissociation constant about 1 nM). Maximal binding of 125I-Ang II to brain microvessels was about 2-fold higher than the maximal binding to particulate fractions of the cerebellum and more than 15-fold higher than that of the cerebral cortex. No significant differences were noted between cerebral and cerebellar microvessels in their specific binding of Ang II. Furthermore, our finding that analogues of Ang II displace specific 125I-Ang II binding to brain microvessels in a rank order that correlates with their pharmacological activities confers biological relevance on the ligand-binding studies. These results strongly suggest that specific Ang II receptor binding sites are present in brain microvessels. Such Ang II receptors may have an important role in regulating the microcirculation of the brain.     

Isolated microvessels: the blood-brain barrier in vitro.

Isolated bovine retinal and brain microvessels, exhibiting a patent lumen, were used to study the contribution of the microvasculature to the blood-brain and blood-retina barriers. The diffusion marker, sucrose, was taken up slowly by the isolated microvessels in contrast to leucine, tyrosine, and valine which were taken up at a considerably faster rate. Uptake of leucine was temperature dependent but resistant to inhibition by ouabain and sodium azide. The large neutral amino acids exhibited stereospecificity and cross-competition for uptake by the isolated microvessels. The apparent Kms for uptake for tyrosine, leucine, and valine were III micron,133 micron, and 500 micron, respectively.     

PDGF and the testis.

Testicular development is controlled by a complex hierarchy of gene regulatory proteins, growth factors, cell adhesion molecules, signaling molecules and hormones that interact, often acting within short time windows, via reciprocal control relationships. The identification in the testis of platelet-derived growth factor (PDGF), a key regulator of connective tissue cells in embryogenesis and pathogenesis, has focused attention on the role of this growth factor in testicular pathophysiology. This review summarizes recent advances in the study of the actions of PDGF in the male gonad, and attempts to incorporate complex in vitro and in vivo experimental data into a model that might clarify the role played by PDGF in the mammalian testis.     

Tissue engineering of perfused microvessels

One major obstacle toward the creation and survival of larger, three-dimensional tissues is the lack of a vascular network that provides transport of oxygen, nutrients, and metabolic byproducts. Although attempts to create microvasculature in vitro have been described previously (Microcirculation 2 (1995), 377; Tissue Eng. 6 (2000), 105; Ann. NY Accd. Sci. 944 (2001), 443), these methods depend on vascularization of void spaces within the tissue-construct or on the utilization of empty capillary networks by host vessels. In the present study, we examined the possibility of creating perfused microvessels in vitro that can be included in an artificial tissue. First, strands of nylon line with their ends fit into microtubing were positioned within small perfusion chambers. Vascular smooth muscle cells (SMCs) were then seeded onto the nylon strands and tubing. The cells multiplied to form concentric layers. Layer thickness was approximately 100 microm after 21 days and 150 microm after 28 days of culture. The lines were then extracted and the chambers connected to a perfusion system. The vessels were continuously perfused with culture medium over 7 days without failure. Artificial microvessels may prove useful in tissue engineering and as models for vascular research.     

Presence of norepinephrine and related enzymes in isolated brain microvessels.

Norepinephrine and the enzymes involved in its synthesis and degradation were found to be associated with isolated brain microvessels. The significance of these results are discussed with respect to adrenergic innervation of the cerebral microvessels and thereby neural regulation of the cerebral microcirculation.     

Impaired myogenic responsiveness of renal microvessels in Dahl salt-sensitive rats.

The mechanisms mediating abnormal renal autoregulation in Dahl salt-sensitive (DS) rats have not been fully defined. In the present study, we assessed myogenic responsiveness of interlobular arteries (ILAs), afferent arterioles (AAs), and efferent arterioles in isolated perfused hydronephrotic Dahl rat kidneys. Dahl rats were divided into four groups according to strain (Dahl salt-resistant [DR] or DS rats) and dietary sodium manipulation (rats fed low or high salt diets). Systolic blood pressure was elevated only in DS rats fed the high salt diet (202 +/- 4 mm Hg, p less than 0.05). Myogenic responses were obtained by stepwise elevation of renal arterial pressure. Vessel diameters were determined by computer-assisted videomicroscopy. Preglomerular microvessels of DS and DR rats responded differently to changes in renal arterial pressure. AAs and ILAs manifested diminished myogenic responsiveness to increasing renal arterial pressure in DS rats compared with DR rats (p less than 0.05). Both AAs and ILAs in DS rats manifested a higher threshold pressure for eliciting myogenic responses and a decrease in maximal pressure-induced vasoconstriction. The sensitivity of the AA myogenic response to nifedipine was enhanced in DS rats compared with DR rats (p less than 0.05). For rats fed the high salt diet, preglomerular vessels exhibited reduced myogenic responsiveness in both strains. In contrast to preglomerular microvessels, efferent arterioles from all four groups of rats failed to exhibit pressure-induced vasoconstriction. Our data suggest that diminished myogenic responsiveness of AAs and ILAs in DS rats contributes to impaired renal autoregulation in this strain.     

Ultrastructure of primary lymphosarcoma of testis.

A case of primary lymphosarcoma of testis was studied by light and electron microscopy. The fine structure of the lymphosarcoma cells was simple and monotonous. The cytoplasm was scanty and contained normal-looking organelles. Free ribosomes were abundant and the Golgi apparatus was well developed. The nuclei were large and irregular in shape with prominent nucleoli. "Nuclear pockets" enclosing material similar to the cytoplasmic ground substance were found infrequently. Remnants of disintegrating cells were also encountered.     

Adenosine 5'-triphosphate induced dilation of human coronary microvessels in vivo.

OBJECT: This study was performed to compare the coronary microvascular response to adenosine 5'-triphosphate (ATP) with the response to adenosine in humans. METHODS: Coronary blood flow velocity was determined using a Doppler flow wire. After intracoronary nitroglycerin infusion, intracoronary bolus injections of adenosine (20 microg) and ATP (20 microg) were performed to induce reactive hyperemia. PATIENTS: Twenty-nine patients (23 men and 6 women, mean age: 63+/-9 years) with coronary artery disease and risk factors for coronary atherosclerosis were studied. RESULTS: Coronary flow reserve in response to ATP was similar to that for adenosine (2.7+/-0.7 vs. 2.7+/-0.7). However, the duration of ATP-induced vasodilation was longer than that of adenosine-induced dilation (39+/-25 seconds vs. 26+/-12 seconds, p<0.0001). The coronary flow reserve obtained with either ATP or adenosine was significantly reduced in the interventioned arteries compared with non-stenosed arteries. The coronary flow reserve obtained with ATP was similar to that obtained with adenosine in both artery groups. The duration of the vasodilator effect of ATP was significantly greater than that of adenosine in both artery groups. CONCLUSION: These results suggest that ATP induces maximal dilation of coronary microvessels, most likely through an endothelium-independent mechanism. The degradation of ATP to adenosine 5'-monophosphate (AMP) and adenosine, as well as the direct action of ATP on A2-adenosine receptors may be responsible for the dilation. We conclude that coronary flow reserve can be determined safely with intracoronary ATP administration.     

Sequence of morphological and hemodynamic changes of gastric microvessels in portal hypertension.

Changes in gastric microvasculature and blood flow at different phases of portal hypertension were studied in rats 1, 2, 3, 4, and 15 days after induction of portal hypertension or sham operation. Vessel lumen and vessel wall thickness were expressed as a ratio referred to the vessel size. On day 2 after constriction of the portal vein, gastric blood flow was decreased (0.57 +/- 0.06 vs. 0.99 +/- 0.20 mL.min-1.100 g-1; P less than 0.05), and gastric vessels had a distended lumen (0.42 +/- 0.02 vs. 0.28 +/- 0.03; P less than 0.01) and a thin wall (2.11 +/- 0.2 vs. 3.82 +/- 0.4; P less than 0.01). On day 4, the gastric blood flow of portal hypertensive animals was increased (1.15 +/- 0.14 vs. 0.71 +/- 0.07 mL.min-1.100 g-1; P less than 0.05), whereas gastric vessels had a reduced lumen (0.27 +/- 0.02 vs. 0.33 +/- 0.02; P less than 0.01) and a thick wall (4.19 +/- 0.52 vs. 3.16 +/- 0.30; P less than 0.05). By day 15, vessels with the largest lumens (0.45 +/- 0.01 vs. 0.29 +/- 0.01; P less than 0.01) and the thinnest walls (1.78 +/- 0.26 vs. 3.58 +/- 0.62; P less than 0.01) were observed in portal hypertensive animals. In conclusion, the gastric vessels of the 15-day portal vein-ligated rat resemble the structural abnormalities described in human portal hypertensive gastropathy.     

A method for measuring the rate of oxygen release from single microvessels.

A system determining the rate of oxygen release from erythrocytes flowing in single microvessels was constructed with an inverted microscope by connecting 1) a scanning/grating spectrophotometer equipped with a photon-counting detector through a thin light guide, to obtain the visible absorption spectrum of a spot (5 microns in diameter) focused on a microvessel, 2) two photomultipliers (connected to a microcomputer via an analog-to-digital converter) through two light guides, to determine the flow velocity of erythrocytes by calculating the cross correlation between the light-intensity changes of two spots (3 microns in diameter, 5 microns apart from each other) focused on the microvessel, and 3) an image processor through a video camera, to estimate the diameter of microvessel from the digitized video images. The rate of oxygen release from single microvessels 7-25 microns in diameter in rat mesentery was measured under the superfusion of deoxygenated solution: 1) The maximal rate was obtained in capillaries, and the rate in arterial microvessels was larger than that in venous microvessels, when similar diameters were compared. 2) The rate was maximum at pH 7.0-7.2, and it decreased in more acidic and alkaline pH values. 3) The rate decreased with a decrease in temperature. The reliability of the measurement using the present apparatus was tested in detail.