Transverse testicular ectopia with persistent Muellerian duct syndrome.

A one-year-old boy was admitted with complaints of swelling in the left inguinoscrotal region and an empty right scrotum since birth. The inguinoscrotum contained two ovoid solid swellings one above the other. The swellings were testes like in feel, size and shape. The diagnosis was transverse testicular ectopia. The case was managed by surgery. Tissue from gonads, the tubular structures and the fallopian tubes were sent for histopathological examination for confirmation. This was a case of an otherwise normal male with transverse testicular ectopia with persistent Muellerian duct syndrome in the left inguinoscrotal hernial sac.

Blood flow decrease induces apoptosis of endothelial cells in previously dilated arteries resulting from chronic high blood flow.

We investigated apoptosis of endothelial cells during the arterial narrowing process in response to reduction in flow. The decrease in flow was created in the carotid artery by closure of an arteriovenous fistula (AVF), which had been established for 28 days in rabbits. The endothelial cell apoptosis in the carotid artery was studied at 1, 3, 7, and 21 days of flow reduction after closure of the AVF by use of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) with laser scanning confocal microscopy and transmission and scanning electron microscopy. After AVF closure, arterial lumen diameter was reduced by 36%, and compared with endothelial cells before the closure, the number of endothelial cells was decreased by 45% at 21 days. Endothelial cell apoptosis was observed at 1 day, peaked at 3 days (381.3+/-87.1 cells per square millimeter), and decreased at 7 days. These cells had irregular protrusions under scanning electron microscopy and were characterized by fragmented nuclei under transmission electron microscopy. Apoptotic cells were mainly beneath the endothelium and were occasionally within smooth muscle cells and endothelial cells. The results suggest that apoptosis of endothelial cells may play a role in the arterial remodeling in response to a reduction in flow.

Molecular mechanisms of aortic wall remodeling in response to hypertension.

OBJECTIVE: The molecular basis of vascular response to hypertension is largely unknown. Both cellular and extracellular components are critical. In the current study we tested the hypothesis that there is a balance between vascular cell proliferation and cell death during vessel remodeling in response to hypertension. METHODS: A midthoracic aortic coarctation was created in rats to induce an elevation of blood pressure proximal to the coarctation. The time course was 1 and 3 days and 1, 2, and 4 weeks for the study of the proximal aorta. Ribonuclease protection assay and Western blot analysis were used to evaluate gene expression of growth and apoptosis-related cytokines with two sets of multiple probes, rCK-3 and rAPO-1. Cell proliferation was determined with BrdU (5-bromo-2'-deoxyuridine) incorporation. Apoptosis was examined with TUNEL (transferase-mediated dUTP nick end-labeling). Morphometry was performed on histologic sections. RESULTS: Coarctation produced hypertension in the proximal aorta, 118 +/- 9 mm Hg versus 94 +/- 6 mm Hg in controls (P <.002). Both messenger RNA and protein levels of transforming growth factor (TGF)-beta1 and TGF-beta3 were increased (P <.005 vs controls). Messenger RNA and protein of Bcl-xS and Fas ligand, known as proapoptotic factors, were both reduced after coarctation (P <.005 vs controls). There was increased BrdU incorporation at 3 days and 1 and 2 weeks (P <.001 vs controls). There were no remarkable changes in the apoptosis rate until 4 weeks later. CONCLUSION: Cell proliferation was stimulated at 3 days, and apoptosis was halted until 4 weeks. These changes were associated with upregulation of TGF-beta and downregulation of Bcl-xS and Fas ligand gene expression. These findings suggest that a coordinated regulation of cell proliferation and cell death contributes to arterial remodeling in response to acute sustained elevation of blood pressure. Cell proliferation precedes apoptosis by 2 weeks in this procedure.

Adaptive remodeling of internal elastic lamina and endothelial lining during flow-induced arterial enlargement.

Gaps in the internal elastic lamina (IEL) have been observed in arteries exposed to high blood flow. To characterize the nature and consequences of this change, blood flow was increased in the carotid arteries of 56 adult, male, Japanese white rabbits by creating an arteriovenous fistula between the common carotid artery and the external jugular vein. The common carotid artery proximal to the arteriovenous fistula was studied at intervals from 1 hour to 8 weeks after exposure to high flow. In the controls, the IEL showed only the usual, small, physiological holes, 2 to 10 microm in diameter. At 3 days, some of the holes in the IEL had become enlarged, but they could not be detected by scanning electron microscopy, despite manifest endothelial cell proliferation. At 4 days, gaps in the IEL appeared as small, luminal surface depressions, 15 to 50 microm wide. At 7 days, the gaps in the IEL had enlarged and formed circumferential, luminal depressions occupying 15+/-5% of the lumen surface. Endothelial cell proliferation persisted in the gaps while proliferative activity decreased where the IEL remained intact. At 4 weeks, as the artery became elongated and dilated, the gaps in the IEL widened as intercommunicating circumferential and longitudinal luminal depressions occupying 64+/-5% of the lumen surface. At 8 weeks, the rate of elongation and dilatation of the artery slowed and the widening of the gaps in the IEL diminished. Endothelial cells covered the gaps throughout. We conclude that flow-induced arterial dilatation is accompanied by an adaptive remodeling of the intima. The gaps in the IEL permit an increase in lumen surface area while endothelial cell proliferation assures a continuous cell lining throughout.

Sequential increases and decreases in blood flow stimulates progressive intimal thickening.

OBJECTIVES: To assess the effect of chronic, repetitive increases and decreases in blood flow on an artery. MATERIALS AND METHODS: Arteriovenous fistulae were created in Japanese male rabbits between the left common carotid artery and the corresponding external jugular vein. Animals were placed into either control groups or one of six cycle groups consisting of flow variations (0.5 cycles, 1.0 cycle, 1.5 cycles, 2.0 cycles, 2.5 cycles and 3.0 cycles). Each complete cycle consisted of 4 weeks of increased flow followed by 6 weeks of normalised flow by fistula ligation. RESULTS: Arteries exposed to increased flow for 4 weeks (0.5 cycles) had a significant increase in lumen diameter without intimal thickening. After 6 weeks of normalised flow (1.0 cycle), shear stress became subnormal (0.42 +/- 0.17 N/m2), intimal thickening developed. In subsequent cycles, intimal thickening continued to develop with each point of flow normalisation and reduction in shear stress. Histologic and ultrastructural analysis revealed endothelial cells preservation at all time points, with individual strata of smooth muscle cell proliferation in the intima corresponding to the cycle numbers. CONCLUSION: Progressive intimal thickening occurred in the previously flow-induced remodelled artery when shear stress was reduced to subnormal levels with preserved endothelium, but was inhibited by high flow periods.

Basic fibroblast growth factor expression precedes flow-induced arterial enlargement.

Arteries enlarge in response to increased blood flow, but the molecular signals controlling this enlargement are not well understood. Basic fibroblast growth factor (bFGF) is a potent mitogen for endothelial cells (EC) and smooth muscle cells (SMC) and promotes cellular proliferation and differentiation. In order to determine whether bFGF is expressed in response to increased blood flow in vivo, carotid-jugular arteriovenous fistulas (AVF) were created in Japanese white rabbits. The carotid artery proximal to the fistula was harvested after 1, 3, or 7 days and compared to nonoperated, control carotid arteries. Arterial blood flow increased five- to eightfold in all AVF animals and resulted in a significant increase in wall shear stress. The proximal carotid artery arterial diameter was no different from control after 1 and 3 days (2.3 +/- 0. 1 mm) but enlarged to 2.9 +/- 0.1 mm (P < 0.05) after 7 days. RT-PCR revealed early transcription of bFGF mRNA at 1 and 3 days with increased densitometric band ratio (bFGF/beta-actin) at 7 days. Immunohistochemical analysis revealed bFGF protein localization in EC of control arteries as well as AVF arteries at all time points. SMC and adventitia expression of bFGF was absent in controls, minimal at 1 day, and increased after 3 and 7 days in the experimental groups. Western blotting confirmed the presence of bFGF in samples and transmission immunoelectron microscopy confirmed its nuclear localization. Endothelial cells in vivo express bFGF under both normal and elevated flow conditions. Smooth muscle cells, however, do not express bFGF under normal flow conditions but begin to express bFGF after 1 day of high flow with increased expression after 3 and 7 days. Flow-induced arterial enlargement begins after SMC expression of bFGF. Therefore, bFGF may play a role in arterial enlargement and adaptive remodeling in response to increased flow.

The effect of hypercholesterolemia on early atherosclerotic lesions initiated by fibrinopeptide B.

Hypercholesterolemia and thrombosis have been implicated as factors in the development of atherosclerosis. Fibrinopeptide B (FPB) is a short chain peptide cleaved from fibrinogen during the production of fibrin. FPB is a known chemoattractant and has been shown to produce experimental atherosclerotic lesions in association with hypercholesterolemia. The present study was designed to examine the role of hypercholesterolemia in this process and to study the time course of the development of these lesions. Twelve New Zealand White rabbits were placed on an atherogenic diet and had suture carrying either FPB, fibrinopeptide A (FPA), or saline (controls) implanted in the adventitia of the femoral arteries and were sacrificed at 14 days. An equal number of animals were left on a standard diet and underwent similar treatment. Eleven animals were treated as the hypercholesterolemic group but were sacrificed at 2, 4, and 7 days. The thickness of the intima was measured adjacent to the suture in the animals sacrificed at 14 days, and the hypercholesterolemic FPB sites were thicker (12.23 mu +/- 6.60) than either hypercholesterolemic FPA (6.06 mu +/- 3.72), saline (4.94 mu +/- 1.42), or the normocholesterolemic FPB (5.99 mu +/- 4.61), FPA (3.89 mu +/- 2.20), or saline (3.97 mu +/- 1.83) (P less than 0.05 for all groups). Transmission electron microscopy of the hypercholesterolemic FPB group showed evidence of macrophages, actively secreting smooth muscle cells with newly deposited elastin, and foam cells by 7 days. We conclude that FPB attracts or stimulates macrophages and smooth muscle cells and that the resultant cellular and extracellular proliferation favors early atherosclerotic lesion formation in the presence of hypercholesterolemia.

Role of fibrinopeptide B in early atherosclerotic lesion formation.

The development of atherosclerotic lesions involves many cell types, including macrophages. Fibrinopeptide B (FPB) has been shown to be a potent chemotactic agent for macrophages, which are abundant as intimal foam cells in atherosclerotic lesions, especially in cholesterol-fed rabbits. We hypothesize that intimal low-density lipoproteins also cause fibrinogen in the intima to release FPB and that FPB attracts macrophages in response to the high lipid levels associated with lesion development. To test our hypothesis, we used an atherosclerotic model. Silk sutures containing either FPB, fibrinopeptide A (FPA), lipopolysaccharide (LPS), or saline control were prepared. One suture of each type was placed in the adventitia of the femoral artery of a rabbit. Animals were killed at 1 or 2 weeks. Only vessels exposed to either FPB or LPS showed significant intimal thickening in the region adjacent to the suture site. Semi-thin electron microscopic sections indicated that the intimal wall was highly cellular and that many cells contained lipid vacuoles after 2 weeks. These sections also showed that the endothelium remained intact and that no injury to the media of the artery had occurred. Electron microscopy of the tissue samples showed the proliferation of smooth muscle cells and deposition of extracellular matrix in the 2-week animals, whereas foam cells were present in the 1-week animals. We conclude that FPB does indeed attract macrophages to the intima and that these macrophages may become foam cells. The model we have developed can be used to study possible mechanisms for the entry of macrophages into the intima during early lesion development and to further understand the complex interactions of FPB, fibrinogen, and lipids in atherosclerotic lesion development.