Our work centers on understanding how the extracellular matrix molecule tenascin-C regulates neuronal growth. We have found that the region of tenascin-C containing only alternately spliced fibronectin type-III repeat D, called fnD, when used by itself, dramatically increases neurite outgrowth in culture. We used overlapping synthetic peptides to localize the neurite outgrowth-promoting site within fnD to a 15 amino acid sequence, called D5. An antibody against D5 blocked promotion of neurite outgrowth by fnD as well as tenascin-C, indicating that this peptide sequence is functional in the context of the native molecule. Further testing of shorter synthetic peptides restricted the neurite outgrowth-promoting site to eight amino acids, VFDNFVLK. Of these,"FD" and "FV" are conserved in tenascin-C sequences derived from all the species available in the GenBank. To investigate the hypothesis that FD and FV are critical for the interaction with neurons, we tested a recombinant fnD protein and synthetic peptides with alterations in FD and/or FV. These molecules did not facilitate process extension, suggesting that the conserved amino acids are required for formation of the active site in fnD. We next investigated whether VFDNFVLK could be used as a reagent to overcome the neurite outgrowth inhibitory properties of chondroitin sulfate proteoglycans, the major inhibitory molecules in the glial scar. The peptide significantly enhanced outgrowth on proteoglycans and was more effective than laminin-1, L1-Fc, or intact tenascin-C, thus demonstrating the potential applicability of tenascin-C regions as therapeutic reagents.

Reactive gliosis, observed in numerous pathological states, leads to the formation of a glial scar that is believed to impede axonal regeneration. Astrocyte reactivity can be initiated both in vitro and in vivo by various cytokines. Thus, the aim of this study was to investigate if suramin, a polysulfonated napthylurea that has been shown to inhibit the binding of many different cytokines to their cell surface receptors, could attenuate the glial response after brain injury. A single dose of suramin (5 microl, 75 microM) or saline vehicle was injected intracerebrally through the same needle used to make the stab wound at the time of lesioning. Suramin-treated animals showed an obvious reduction in several parameters of CNS inflammation: cellular proliferation, GFAP levels, and tenascin-C immunoreactivity were reduced in suramin-treated as compared to control animals at early time points. GFAP immunoreactivity was strikingly reduced at 3 days after injury, as confirmed by Western blot analysis. This reduction was transient, however, in that the difference in GFAP expression between suramin-treated and control animals was less apparent at 7 days and had disappeared by 30 days after injury. Likewise, fewer BrdU-positive cells were noted in treated versus control tissue at 1 and 3 days, but this difference was not significant by 7 days. Moreover, tenascin immunoreactivity was significantly diminished at 24 h as confirmed by Western blot analysis in suramin-treated lesion areas, which is analogous to our observations that suramin can antagonize tenascin expression by cultured astrocytes treated with bFGF. In addition, examination of the corpus callosum of saline-treated animals 30 days post-trauma revealed a disruption of the fiber tract within the lesion site, while suramin-treated animals displayed numerous fibers spanning the lesion. These results demonstrate that a single injection of suramin transiently inhibits the gliotic response, which may be sufficient to ameliorate subsequent tissue damage.

Hepatocyte growth factor (HGF) has unique morphogenic activity for several cell types. Besides its major effect upon liver regeneration, its motogenic activity to enhance motility has not been verified for smooth muscles. Therefore we evaluated the impact of HGF in an in-vitro model of human gallbladder motility. Twelve stone-diseased and eight stone-free muscle strips were preincubated with HGF (100 ng/ml, 200 ng/ml). For the analysis of motility, cholecystokinin (CCK) was added (0.1 nM, 0.5 nM, 2 nM, 10 nM, and 100 nM). Twelve stone-diseased and eight stone-free strips without HGF incubation served as the control group. The tone of healthy (tone/100 nM CCK: control group, 12.4 +/- 3.6 mN; HGF group, 19.5 +/- 4.5 mN) and stone-diseased (tone/100 nM CCK: control group, 10.8 +/- 3.8 mN; HGF group, 17.3 +/- 4.8 mN) muscle strips, preincubated with HGF, was increased, with a higher sensitivity to CCK. Our results suggest that there is a clear motogenic response of stone-diseased human gallbladders to HGF.

Constriction of the aorta just above the origin of both main renal arteries invariably resulted in elevation of the carotid systolic and carotid mean pressure. The hypertension was not immediate, but developed in about the same time as after constriction of the main renal arteries (3). Constriction of the aorta just below the origin of both main renal arteries had no significant effect on the carotid systolic or carotid mean pressure. Since these results were first reported (1), Rytand (88, 89) has shown by an indirect method, namely, the demonstration of the development of cardiac hypertrophy, that hypertension in the upper part of the body can be produced in the rat by constriction of the aorta just above the origin of both main renal arteries. The immediate effect of constriction of the aorta either below or above the main renal arteries is a fall of blood pressure (femoral mean pressure) below the site of the clamp, the extent of the fall being directly dependent upon the degree of constriction of the aorta. Of particular interest is the eventual elevation of the femoral mean pressure above the normal in some animals with the aorta constricted or even occluded above the origin of the main renal arteries. This was most pronounced and persistent in those animals in which, in addition, the aorta below the origin of the renal arteries, and, in some animals, the main renal arteries, also were constricted. The most important factors which determined this elevation of blood pressure in the lower part of the body were probably increased flow of blood into the vascular bed below the clamp and peripheral vasoconstriction of renal and humoral origin, as in the case of the hypertension produced by constriction of the main renal arteries alone (2-86). Although elevation of the carotid systolic or carotid mean pressure occurred invariably within 24 to 48 hours after the constriction of the aorta above the site of origin of both main renal arteries, yet there was a tendency, after a variable period, for the elevated blood pressure to become lower or even to drop to the original level. Increased constriction, and finally occlusion of the aorta, above the origin of the main renal arteries, and even constriction or occlusion of the aorta below the renal arteries, in addition, failed to induce hypertension that persisted for a long time at a high level. In order to produce this effect, it was necessary to constrict the main renal arteries as well. The possible explanation of the failure of the hypertension to persist for a long time after constriction of the aorta alone, is that the initial ischemia of the kidneys disappeared due to the improvement of the blood flow through the kidneys as a result of (a) the increase of the natural accessory circulation to the kidneys;(b) the increased blood pressure above the site of the clamp and consequent increased flow of blood into the part of the aorta below the clamp;(c) increased pressure below the site of the clamp due, in great part, to peripheral vasoconstriction, and in part to the increased inflow of blood into the lower part of the body through the aorta and collateral channels. For the dog, this method is not necessary for the production of persistent hypertension. Constriction of the main renal arteries is easily performed and is effective for the production of generalized hypertension (2-11). However, constriction of the aorta in addition to constriction of the renal arteries results in greatly elevated persistent hypertension. Constriction of the aorta alone above the origin of the main renal arteries would be useful in the dog only for the production of relatively short periods of hypertension in the upper part of the body. For small animals it may be a more effective and useful method. In the dog, the only technical difficulty encountered was the erosion of the wall of the aorta by the clamp. This may not occur in small animals. In previous studies (2-11) that have dealt with the constriction of the main renal arteries, this accident rarely occurred. When the constriction of the aorta above the origin of the main renal arteries was of moderate degree, or was gradually made very great, the resultant hypertension was not accompanied by impairment of renal excretory function, as determined by urea clearance or by the quantity of urea, creatinine or non-protein nitrogen in the blood, the benign phase of hypertension (3). When the constriction of the aorta was suddenly made very great, impairment of the renal excretory function usually followed, and the animal developed fatal convulsive uremia and characteristic vascular lesions, the malignant phase of hypertension (9). These facts, are all indicative of the renal origin of the hypertension which results from the constriction of the aorta just above the origin of both main renal arteries. Hypertension did not persist for a sufficiently long time to permit any conclusive comparison between the effect of the high and low pressures on the structure of the vascular system, above and below the site of the clamp, respectively. During the period of survival of these animals, no significant differences were observed between the appearance of the vascular system of the upper part of the body and that of the lower part of the body, and significant cardiac hypertrophy did not develop. In the aorta and large arteries, intimal arteriosclerosis was not observed. In the aorta of one old animal several small plaques of calcification were found in the media, but these were present in the portion of the aorta below, as well as above the clamp, and they were no larger or more abundant than were observed in some old dogs with normal blood pressure. Dogs 3-50 and 3-83, that are still alive, with very high blood pressure above the site of the aortic clamps, and relatively low pressure (though greater than normal) below the site of the aortic clamps, will be valuable for the determination of possible differences between the effects of the two levels of blood pressure in the large and small blood vessels. In these dogs also, it will be possible to determine the effect of the persistently high blood pressure on the myocardium. The possible application of the results of this study to the problem of the pathogenesis of human eclampsia is mentioned here for consideration. Since this condition occurs in pregnancy only at a time when the uterus is greatly enlarged, it is at least possible that the mass may press on the aorta or both main renal arteries sufficiently to produce renal ischemia. The suddenness with which the uremic convulsive phase of eclampsia develops is in keeping with this idea. In the dog, an aggravating effect of pregnancy on an already established hypertension has not been noted. As a matter of fact, most of the hypertensive dogs that have become pregnant, have shown a slight or moderate fall, rather than an increased rise of pressure. Since the dog stands with the body in a horizontal position, and does not lie on its back, pressure of the pregnant uterus on the aorta and blood vessels is less than in human beings who stand erect and frequently lie on their backs. The soundness of this suggestion could be tested by placing pregnant women, in the early stage of eclampsia, in a position which could relieve possible pressure on the aorta and main renal arteries. A possible explanation of the fall of pressure in the pregnant hypertensive dogs is the compensatory effect of the normal kidneys of the pups, as in the case of an animal with one main renal artery constricted and the other kidney normal. As has been shown (3, 31, 72), the presence of one normal kidney in an animal hypertensive due to constriction of the other main renal artery, results, after a variable period, in a return of the blood pressure to normal. How the normal kidney acts to produce this effect is not known.

When faradic stimulation was undertaken of vessels irrigated with Ringer's solution, which alternately contained and was free from carbon dioxide, it was observed that the reaction was far less when the solution contained carbon dioxide. A reversal of the effect could be obtained many times. It appears, therefore, that when Ringer's solution contained carbon dioxide in the concentration described, the irritability of the vessels to electrical stimuli decreased, although carbon dioxide by itself and in the absence of the application of the stimuli, appeared to be void of effect upon the vessels. The rare, divergent results were traced to technical errors. We attempted to discover whether the observed decrease in irritability of the vessels might not be due to the absence of oxygen. For this purpose we irrigated the vessels with Ringer's solution alternately containing nitrogen and oxygen. When nitrogen caused any change this was due to an influence on the rate of the heart and not on the irritability or reactivity of the arteries. In whatever way we tried we were unable to bring about a change in reactivity of the arteries by creating a condition of oxygen lack independently of a change in the rate of the heart beat. We attempted to study also the effect of other acids beside carbon dioxide on the changed reactivity of the arteries. Irrigation with various concentrations of lactic acid was without result. We also employed solutions buffered with potassium and sodium phosphate. When the irrigation was undertaken with these solutions having a pH range varying from 7.7 to 5.9 we observed neither a direct action nor one which modified the preparation in such a way as to change its susceptibility to faradic stimulation. Important investigations have been published recently by Atzler and Lehmann (2) on the direct influence of the hydrogen ion concentration on the behavior of blood vessels. Hammett and Zoll believed that, as the result of their experiments in which they attempted to bring about stimulation with solutions of concentrated carbon dioxide, they were able to exclude the possibility of action due to acid alone and therefore ascribed to carbon dioxide a specific effect. In our own experiments the method of irrigation does not permit an inference whether, or how far, an acid effect plays a rôle in the carbon dioxide experiments. For beside the question of hydrogen ion concentration and of buffering, the question of the penetration of substances from the surface to the contractile elements of the wall of the vessels requires to be considered. Carbon dioxide has an ability, beyond that of all other substances, to penetrate through tissues (3). It may be owing to this property that we could influence the reactivity of the blood vessels with it and it alone. This possibility must be further investigated. In these experiments, however, it was our object to show only that it was possible to influence the irritability of blood vessels experimentally. The conclusion is justified by our experiments that carbon dioxide in small concentrations reduces the threshold of irritability for electrical stimuli of the blood vessels of the embryonic membrane.

On the basis of the anatomical studies presented the following inferences or conclusions are drawn. 1. In the course of development there appear in the vascular membranes of chick embryos arterial vessels of all calibers, namely, capillaries, small arteries with 2 or 3 cell layers, and large arteries formed of endothelium, longitudinal and circular layers of muscle and adventitia. 2. In none of the stages are elastic fibers developed. Only in the most central portion of the umbilical artery, in that portion namely which is to be regarded as belonging to the embryo, are elastic fibers discoverable. 3. The structure of capillaries is histologically the same at all stages. The small arteries of embryos 10 days old resemble histologically those of 18. At no stage of development are appearances of degeneration nor of fat to be found in arteries. When the physiological results of our investigations are compared with the anatomical ones the following comments may be made. In respect to Paragraph 1 of the anatomical results we may remark that when we study the different forms of the wall of arterial vessels the most delicate vessels consisting of single cells exhibit the greatest irritability. Those which are built of 3 to 4 muscle layers are less irritable. Stouter vessels appearing for the first time at 10 days of incubation require stronger stimuli to bring about the same reaction. In respect to Paragraph 2 of the anatomical results we may make this comment. The absence of elastic fibers in all arteries of the embryonic membranes throughout the period of their development is important in defining a physiological property of the larger vessels. The medium and larger vessels, beginning with the 4th day of incubation, contract differently from normal adult human arteries. In the contracted state they appear in cross section not as small replicas of larger circular structures, but take on a new form. During the course of contraction they become flat and appear band-like as would a garden hose when it is compressed by a weight. In examining a vessel so contracted one sees on rotating the vessel either a broad side or a narrow one. It is for this reason that such arteries appear alternately narrow as a line or broad as a band. It is not until the narrow artery is elevated with a hook that its uniform band-like nature becomes evident. The absence of elastic tissue, the presence of which in all probability is mainly responsible for the usual shape of arteries on cross section, permits one to see how the phenomenon which has been described may come about. Concerning Paragraph 3 of the anatomical conclusions we have this to say. According to the histological investigation a stage of degeneration is wanting in the blood vessels of the embryonic membrane in a sense in which one is accustomed to see such changes in other blood vessel systems during the course of life. On the day of hatching the constituent cells and fibers of the arteries of all calibers are anatomically the same as in their early development. These vessels do not die as the result of aging: The nutrient fluid ceases to flow because of contraction of the umbilical vessels. The blood vessels die in complete possession of their physiological irritability and anatomical integrity. The unaltered irritability of blood vessels of the same caliber at all ages is consonant with their unaltered anatomical structure.

1. The allergic irritability of the guinea pig (capacity of the animal to react to antigenic substances) is increased by infection with Bacillus abortus and a streptococcus, by the dead tubercle bacillus, and by intensive treatment with trypan blue, respectively. The effect of these influences, while definite, is less pronounced than that previously found for infection with the tubercle bacillus. The production of anti-sheep hemolytic amboceptor was used as the test reaction in these cases. 2. The allergic irritability of the guinea pig with reference to anti-typhoid agglutinin is increased by infection with the tubercle bacillus. 3. The allergic irritability of the rabbit with reference to anti-sheep hemolytic amboceptor is increased by an infection of suitable severity with the tubercle bacillus. 4. In the guinea pig the curve of antibody production is complex. Its peculiarities are developed during the production of antityphoid agglutinins as well as that of anti-sheep hemolytic amboceptor. In the latter case injections of antigen subsequent to the first give rise to a curve of production unchanged in form but somewhat affected in the time relations. 5. The effects of infection with Bacillus tuberculosis on allergic irritability with reference to anti-sheep hemolytic amboceptor are operative throughout a course of immunizing treatments. The successive increases due to the cumulative effect of repeated doses of the antigen are developed on a higher level. The end-result is that the animal with increased irritability furnishes more antibody not only in response to the initial injection of antigen as previously described, but an absolute increase over the amount attainable by a comparable number of treatments in series. That portion of the final result contributed by the increase in allergic irritability appears to be no less, and may even in instances be somewhat more than that due to the earlier doses of the specific antigen.

OBJECTIVE: To investigate the embolization effect of Pingyangmycin-albumin microspheres (PYM-AMS) on small arteries and its process of degradation in vivo. METHODS: Twenty four Japanese white rabbits were randomly divided into four groups, 6 in each group. PYM hydrochloride + 0.9% NaCl, PYM + soybean oil, and PYM-AMS + soybean oil were injected into the central auricular arteries of the rabbits in the three experimental groups, respectively, for about 30 seconds (0.26 mL/per ear, which contained PYM 5 mg/mL). The vessel samples were taken and examined at 2, 7, 14, and 21 days. RESULTS: The PYM + 0.9% NaCl group had no significant vessel changes. In the PYM+ soybean oil group, some endothelial cells dropped off at the 7th day after injection. At the 21st day, mild proliferation of endothelial cells and walls of central auricular arteries were observed, especially on the intima. But the lumen was still obvious and the blood flow was not blocked. In the PYM-AMS group, the central auricular arteries were narrowed at the 7th day after injection. At the 21st day, the vessels had sclerostenosis, and the blood flow was blocked. At the 14th day, significant proliferation of endothelial cells and walls of central auricular arteries were observed. The surface of PYM-AMS was absorbed. At the 21st day, the walls of central auricular arteries and some small veins proliferated obviously, and the arteries were sclerostenosed. Many smooth muscle cells, fibroblasts, and myofibroblasts in the original blood vessel lumen appeared. There were thrombi besides the PYM-AMS. CONCLUSION: PYM-AMS may become an option for the treatment of large venous or arteriovenous malformations and for the local chemotherapy of malignant tumors.

Treatment of angiospasm as one of the most dangerous complications of subarachnoidal hemorrhage in the cerebral arterial aneurysm rupture constitutes an actual problem. Possibility of the cerebral vessels visualization have permitted to use intraarterial infusion of vasoactive preparations (IAIVAP) for treatment of angiospasm. Therapeutic efficacy for angiospasm, cerebral ischemia and edema depends on many factors and even in application of IAIVAP not always terminates successfully. The disease predictors studying makes possible to plan the intensive therapy tactics correctly, to prognosticate the disease course and the result of treatment. While analyzing the material there were used statistical methods of the data processing, permitting to reveal the prognostically meaningful criterions, influencing the treatment efficacy for angiospasm and its consequences. There was proved the influence of some predictors on the survival and mortality indexes, as well as on disablement and securing of ability to work in patients after application of IAIVAP. It was established, that conduction of IAIVAP had promoted the trustworthy raising of the survival indexes, as well as the lowering of mortality and the disablement rate in patients suffering angiospasm, cerebral ischemia and edema.

Atherosclerosis in a major leg artery leads to impaired blood supply, which normally progresses to critical limb ischemia. Atherosclerosis produces substantial alterations of structure and endothelial function in the large conduit arteries. Pressure unloading and ischemia in the distal vasculature bring about alterations in microvascular function. Resistance arteries undergo significant wall thinning and changes in their contractile regulation. Optimization of large artery dimensions by the small arteries through flow-mediated vasodilation is impaired. Angiogenesis is stimulated, which can result in the formation of major collateral feeder vessels in addition to small nutritive blood vessels. However, angiogenesis can also contribute to instability of atherosclerotic plaques, which ultimately leads to further deterioration in blood supply. Surgical bypass grafting to restore blood supply to the distal leg generates a sudden increase of pressure in the weakened resistance vasculature, leading to uncontrolled changes in capillary hydrostatic pressure, extravasation of fluid, and tissue edema. This review aims to highlight the importance of the resistance vasculature in critical limb ischemia and the interdependence of pathophysiological changes in the large conduit and small resistance arteries. The major unresolved question is why the physiological mechanisms that regulate vascular structure and function ultimately break down, leading to circulatory failure within the distal limb.

To elucidate compositional changes of the rami of the internal iliac artery with aging, the authors investigated age-related changes of the calcium content in the uterine, internal pudendal, umbilical, and obturator arteries by inductively coupled plasma-atomic emission spectrometry. After an ordinary dissection was finished, the uterine, internal pudendal, umbilical, and obturator arteries were resected from 10 female subjects, and the internal pudendal, umbilical, and obturator arteries were resected from 10 male subjects. The female subjects ranged in age from 52 to 96 yr, and the male subjects ranged in age from 63 to 88 yr. The calcium content in the uterine artery began to increase in the seventies and increased markedly in the nineties. In the internal pudendal artery, the calcium content hardly increased up to the eighties and increased significantly in the nineties. In contrast, the calcium content did not change in both the umbilical and obturator arteries with advancing age. It was found that the average content of calcium was the highest in the uterine artery and decreased in the order internal pudendal, umbilical, and obturator arteries. The average content of calcium in the uterine arteries corresponded to 46-fold the amount of the women's obturator arteries, in which it was the lowest. In the cases of men, the average content of calcium was higher in the order of the internal pudendal, umbilical, and obturator arteries. Regarding the average content of calcium, the order internal pudendal, umbilical, and obturator arteries of the men was consistent with that of the women.

BACKGROUND/AIMS: The main aim of the study was to evaluate whether superior hypogastric plexus and hypogastric nerve can be preserved without increasing local recurrence while performing surgical treatment of rectal carcinoma. METHODOLOGY: This was a retrospective study of 129 patients with rectal carcinoma who underwent curative resection with two types of autonomic nerve-sparing operation. The superior hypogastric plexus and bilateral hypogastric nerves were resected in 61 patients and spared in 68 patients. The pelvic plexus was preserved in all the patients. Local recurrence and survival were compared between two operations. RESULTS: After three years, local recurrence cumulative rates were 13.1% after hypogastric nerve removing operation and 10.3% after hypogastric nerve preserving operation. Distant metastasis and corrected 5-year survival rates were 23.0 and 61.6%, respectively after hypogastric nerve-removing operation, while after hypogastric nerve-preserving operation those were 16.2 and 77.4%, respectively. There were no statistically significant differences in local recurrence, distant metastasis and survival between the two groups. CONCLUSIONS: Hypogastric nerve-preserving operation does not appear to carry an increased risk of local recurrence compared with hypogastric nerve-removing operation after an equivalent follow-up period.