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Biomedical subjects

P Seifert

Publications and source records attributed to P Seifert.

At least 19 recordsLinked to original sources

Histopathologic evaluation of aneurysms treated with Guglielmi detachable coils or matrix detachable microcoils.

BACKGROUND AND PURPOSE: The purpose of this study was to evaluate the degree of organization and fibrocellular tissue development in aneurysms treated with bare platinum or biologically active microcoils. METHODS: Twelve aneurysms were removed at autopsy between 1-18 days and another 2 between 2-3 months posttreatment. Four aneurysms were surgically removed between 6 months and 3 years following treatment. One aneurysm removed at 8 days and another at 6 months were treated with bioactive (Matrix) coils; the other 16 with bare platinum (Guglielmi detachable coils; GDCs). All specimens were embedded in plastic, stained with hematoxilin-eosin and elastin and examined by light microscopy. RESULTS: All specimens removed within 3 weeks demonstrated intra-aneurysmal thrombus, without signs of organization or fibrotic tissue formation over the neck regardless of the type of coils used. In the GDC-treated aneurysms, evidence of early thrombus organization was observed within 2-3 months, and completed yet imperfect fibrocellular reaction together with residual thrombus at 2-3 years. In the Matrix-treated specimens, the aneurysm cavity was completely filled with granulation tissue corresponding to still ongoing fibrocellular reaction at 6 months, including newly formed blood vessels, smooth muscle cells, and collagen deposition without signs of residual thrombus. CONCLUSIONS: Our results indicate that in aneurysms treated with bare platinum coils thrombus organization does not occur until late after treatment and may remain imperfect for years. In one aneurysm studied 8 days following treatment with Matrix coils, no difference was noted compared to aneurysms treated with bare platinum coils. In another aneurysm examined 6 months following packing with Matrix coils, the histologic changes support the hypothesis that the biologically active polymer may accelerate aneurysm healing.

Adolescent↗

An implanted hamster greene melanoma expressing multiple host-tissue differentiation.

After Greene melanoma tissue had been implanted subcutaneously in a Syrian Golden Hamster and allowed to grow there for 10 days, its tissue components were examined by transmission electron microscopy. Amelanotic and melanotic tumor cells, blood vessels and nerve fibers were present. In the amelanotic and melanotic tumor cells the rough endoplasmic reticulum was colonized by spherical endogenous retroviruses, whereas no virus particles could be detected in endothelial or other cells. The cell-type-specific viruses thus mark separate cell lineages and indicate that blood vessels in Greene melanoma are not formed by tumor cells, so that the 'vasculogenic mimicry' currently under controversial discussion for other melanomas is not involved here. The tumor also comprised smooth muscle cells, skeletal muscle fibers and collagen fibers. This remarkable finding implies that tumor growth involves not only the stimulation of angiogenesis and neurogenesis by various growth factors, but also the activation of other cells/tissues in the adjacent host tissues. This influence of the surrounding host tissue produces the observed epiphenomena and can also cause the genotypic character of a tumor to be phenotypically masked (mimicry).

Animals↗

Gold-coated NIR stents in porcine coronary arteries.

BACKGROUND: As endovascular stents are altered to add functionality, eg, by adding radiopaque coatings, biocompatibility may suffer. METHODS AND RESULTS: We examined the vascular response in porcine coronary arteries to stainless steel gold-coated NIR stents (7-cell, Medinol, Inc). Stents, 9 and 16 mm in length, were left bare or coated with a 7-microm layer of gold. Physical and material effects were examined in four different gold-coated stent types, two at each length that either had the coating applied to the standard strut, ie, gold coated thicker than controls, or had the coating applied to thinned struts, ie, gold coated of the same thickness as control struts. Simple gold coating exacerbated intimal hyperplastic and inflammatory reactions over 28 days, but postplating thermal processing smoothed the coating surface and negated the adverse tissue response to gold. The relative amounts of base steel and gold coating and their resistances to expansion and collapse determined the extent of stent recoil. CONCLUSIONS: Gold coatings enhance the radiopacity of steel stents, but not without effects on vascular repair. Material effects predominate and can be abrogated by heating coated stents to alter surface finish and material purity. Clinical results may suffer unless consideration is given to material and physical effects of gold.

Animals↗

Topological-histological investigation of the pterygium.

BACKGROUND: It is a wide-spread assumption, never proven scientifically, that the pterygium is a duplication of the conjunctiva, with an intervening gap at the limbus. We therefore conducted a histological reinvestigation, primarily to clarify the topological relation between normal bulbar tissue and lesion. METHODS: Excised pterygia were prepared for light microscopy and embedding in paraffin. Two pterygia were serially sectioned, and samples of 49 others were removed, sectioned and stained (stains: H&E, alcian blue, toluidine blue, PAS; antibody: cytokeratin 18). RESULTS: The pterygium is an epithelium-covered protuberance of connective tissue, projecting over the normal surface of the eyeball; it consists of a base, which extends in the direction of growth, and lateral lobes. It is therefore impossible to insert a probe all the way under a pterygium. The bulbar tissue adjacent to the pterygial protuberance can likewise be histologically altered. That is, a narrow layer of connective tissue, well supplied with capillaries, may be present in the corneal section of the pterygium between Bowman's layer and the epithelium. CONCLUSION: Histological alterations of the stroma predominate quantitatively over those of the epithelium. The topographical anatomy of the lesion and the fact that bulbar tissue is modified along with the pterygium should be taken into account when excision is undertaken. We predict that if all the altered tissue is removed, the rate of recurrence will be reduced.

Aged↗

Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells.

BACKGROUND: Ultraviolet radiation is thought to play a causative role in various ocular diseases such as macular degeneration, cataract, and possibly melanomas. Since most of the energy is absorbed by pigmented cells, the aim of this study was to examine and compare the reactions of different ocular melanocytic cells to ultraviolet light in vitro. MATERIALS AND METHODS: Bovine iris melanocytes, choroidal melanocytes, iris pigment epithelial cells, and retinal pigment epithelial cells were isolated and cultured. Semiconfluent cultures were exposed to ultraviolet radiation (280-380 nm). Cell number and melanin content were measured 10 days after radiation. Selected samples were examined by transmission electron microscopy. RESULTS: Following irradiation with ultraviolet light for 30 s, 60 s, and 120 s, the number of cells in culture decreased markedly. In contrast, total melanin content in the cultures of iris melanocytes, choroidal melanocytes, and iris pigment epithelial cells did not decrease despite the reduced number of cells. This finding suggested an increase in melanin per cell. However, the increase in average melanin content observed was not due to melanogenesis, because treatment with the melanogenesis inhibitor alpha-methyl-p-tyrosine did not reduce the melanin content of the cultures and electron-microscopic examination showed no morphological evidence of increased melanogenesis. CONCLUSION: In vitro, there was no convincing evidence of ultraviolet radiation-induced melanogenesis in ocular pigmented cells. Thus, it seems that ultraviolet radiation is a selection factor: more densely pigmented cells survive the treatment better than less pigmented cells.

Animals↗

Tumours may be innervated.

It is generally assumed that tumours are not innervated. However, following an accidental observation of a nerve fibre within an adenoma of the ciliary body epithelium of the eye, we have further examined two such tumours. One pigmented and one non-pigmented adenoma of the ciliary body epithelium (APCE and ANCE, respectively) that had been surgically removed from two human eyes were processed for ultrastructural evaluation and systematically screened and analysed for the occurrence of nerve tissue under a transmission electron microscope. The adenomas were composed of epithelial tumour cell strands and interposed vascularised connective tissue. Both tumours contained a small number of fine unmyelinated nerve fibres containing clear and dense core vesicles. In both adenomas, the nerve fibres were located in the tumour periphery close to blood vessels and tumour cells. In the APCE, they were also seen in more central areas. Since nerves always have a function, this finding, if confirmed in other neoplasms, may influence our understanding of such innervated tumours.

Adenoma↗

Glucagon-like peptide 2: a new treatment for chemotherapy-induced enteritis.

BACKGROUND: Glucagon-like peptide 2 (GLP-2) is a recently identified intestinal epithelium-specific growth factor that has been shown to reduce the severity of inflammatory disorders of the intestine in rodent models. We hypothesized that GLP-2 administration would be beneficial in chemotherapy-induced enteritis either by preventing injury or by promoting recovery. MATERIAL AND METHODS: Rats received no drug (control), chemotherapy alone [5-fluorouracil (5-FU), 190 mg/kg, ip] (Chemo), 5-FU followed by 3 days of GLP-2 analog (ALX-0600, 0.1 microg, sc twice daily) (CH-G), or GLP-2 analog for 6 days prior to 5-FU and for 3 days afterward (G-CH-G). Animals were pair fed. Rats received 5-bromo-2-deoxyuridine (Br-dU, 50 mg/kg, 2.5 h prior to sacrifice on Day 3 postchemotherapy) for immunohistochemical assessment of cellular proliferation. RESULTS: Chemotherapy induced significant reductions in body weight, villus height, and crypt depth compared with controls. Intestinal wet weight, villus height, and crypt depth were significantly higher for the CH-G group compared with the Chemo group. The CH-G group also showed a significant improvement in villus height compared with the G-CH-G group. Crypt depth, but not jejunal wet weight or villus height, was significantly improved in the G-CH-G group compared with the Chemo group. The percentage of Br-dU-labeled cells in the intestinal crypts did not differ among the groups. CONCLUSIONS: These results suggest, for the first time, that GLP-2 treatment initiated after chemotherapy administration enhances intestinal recovery. In contrast, GLP-2 treatment initiated prior to chemotherapy administration to prevent injury has less beneficial effect. GLP-2 administration may be beneficial to patients suffering from chemotherapy-induced enteritis.

Animals↗

Stent and artery geometry determine intimal thickening independent of arterial injury.

BACKGROUND: Clinical trials show that larger immediate postdeployment stent diameters provide greater ultimate luminal size, whereas animal data show that arterial injury and stent design determine late neointimal thickening. At deployment, a stent stretches a vessel, imposing a cross-sectional polygonal luminal shape that depends on the stent design, with each strut serving as a vertex. We asked whether this design-dependent postdeployment luminal geometry affects late neointimal thickening independently of the extent of strut-induced injury. METHODS AND RESULTS: Stainless steel stents of 3 different configurations were implanted in rabbit iliac arteries for 3 or 28 days. Stents designed with 12 struts per cross section had 50% to 60% less mural thrombus and 2-fold less neointimal area than identical stents with only 8 struts per cross section. Sequential histological sectioning of individual stents showed that immediate postdeployment luminal geometry and subsequent neointimal area varied along the course of each stent subunit. Mathematical modeling of the shape imposed by the stent on the artery predicted late neointimal area, based on the re-creation of a circular vessel lumen within the confines of the initial stent-imposed polygonal luminal shape. CONCLUSIONS: Immediate postdeployment luminal geometry, dictated by stent design, determines neointimal thickness independently of arterial injury and may be useful for predicting patterns of intimal growth for novel stent designs.

Animals↗

Light and electron microscopic examination of endothelial cells from bovine retinal vessels in long-term cultures.

The aim of the present work was to examine and compare the ultrastructure of bovine retinal endothelial cells (BRECs) in vitro during several passages in a medium selective for endothelial cells. The identity of the endothelial cells was confirmed immunohistochemically, up to the tenth passage. Changes in their ultrastructure in comparison to endothelial cells in vivo occurred at the onset of culturing and not progressively with repeated passages. The cultured BRECs show high metabolic activity in all passages. While retaining their identity as endothelial cells, they modify their lipid metabolism, so that lipids are stored. This change in lipid metabolism was induced by the medium.

Animals↗

Neointimal thickening after stent delivery of paclitaxel: change in composition and arrest of growth over six months.

OBJECTIVES: The purpose of this study was to determine long-term effects of stent-based paclitaxel delivery on amount, rate and composition of neointimal thickening after stent implantation. BACKGROUND: Paclitaxel prevents vascular smooth muscle cell proliferation and migration in vitro and in vivo. These actions, coupled with low solubility, make it a viable candidate for modulating vascular responses to injury and prolonged effects after local delivery. We asked whether local delivery of paclitaxel for a period of weeks from a stent coated with a bioerodible polymer could produce a sustained reduction in neointimal hyperplasia for up to six months after stenting. METHODS: Stainless steel stents were implanted in the iliac arteries of rabbits after endothelial denudation. Stents were uncoated or coated with a thin layer of poly(lactide-co-sigma-caprolactone) copolymer alone or containing paclitaxel, 200 microg. RESULTS: Paclitaxel release in vitro followed first-order kinetics for two months. Tissue responses were examined 7, 28, 56 or 180 days after implantation. Paclitaxel reduced intimal and medial cell proliferation three-fold seven days after stenting and virtually eliminated later intimal thickening. Six months after stenting, long after drug release and polymer degradation were likely complete, neointimal area was two-fold lower in paclitaxel-releasing stents. Tissue responses in paclitaxel-treated vessels included incomplete healing, few smooth muscle cells, late persistence of macrophages and dense fibrin with little collagen. CONCLUSIONS: Poly(lactide-co-sigma-caprolactone) copolymer-coated stents permit sustained paclitaxel delivery in a manner that virtually abolishes neointimal hyperplasia for months after stent implantation, long after likely completion of drug delivery and polymer degradation.

Angiogenesis Inhibitors↗

Adenoma of the pigmented ciliary epithelium: ultrastructural and immunohistochemical findings.

We report the clinical and histological findings in a tumor of the pigmented ciliary epithelium. The tumor was detected because it had caused a unilateral cataract, and it was removed by local resection because a malignant melanoma could not be excluded. The diagnosis was established by light microscopy, and additional immunohistochemical and detailed ultrastructural studies were performed. The so-called foam cells which are considered typical of these adenomas appear to be mostly light microscopic artifacts and had no ultrastructural equivalent in the sections examined from our tumor. Our results also strongly support the hypothesis that no true glandular elements are formed and that, apart from the neoplastic tissue architecture, most of the pathological findings are related to melanosomes.

Adenoma↗

Decreased neointimal formation in Mac-1(-/-) mice reveals a role for inflammation in vascular repair after angioplasty.

Inflammation plays an essential role in the initiation and progression of atherosclerosis, but its role in vascular repair after mechanical arterial injury (i.e., percutaneous transluminal coronary angioplasty, PTCA) is unknown. In animal models of vascular injury, leukocytes are recruited as a precursor to intimal thickening. Furthermore, markers of leukocyte activation - in particular, increased expression of the beta2-integrin Mac-1 (alphaMbeta2, or CD11b/CD18), which is responsible for firm leukocyte adhesion to platelets and fibrinogen on denuded vessels - predict restenosis after PTCA. To determine whether Mac-1-mediated leukocyte recruitment is causally related to neointimal formation, we subjected mice lacking Mac-1 to a novel form of mechanical carotid artery dilation and complete endothelial denudation. We now report that the selective absence of Mac-1 impairs transplatelet leukocyte migration into the vessel wall, reducing leukocyte accumulation over time. Diminished medial leukocyte accumulation was accompanied by markedly reduced neointimal thickening after vascular injury. These data establish a role for inflammation in neointimal thickening and suggest that leukocyte recruitment to mechanically injured arteries may prevent restenosis.

Angioplasty, Balloon↗

Vasoactive intestinal polypeptide (VIP) innervation of the human eyelid glands.

This study was conducted to obtain morphological proof of innervating nerve fibres in the glands of the human eyelid (accessory lacrimal glands of Wolfring, meibomian glands, goblet cells, glands of Zeis, glands of Moll, sweat glands, glands of lanugo hair follicles) and identification of the secretomotorically active neuropeptide vasoactive intestinal polypeptide (VIP) as a common transmitter. Epoxy-embedded ultrathin sections of tissue samples from human eyelids were studied using electron microscopy. Paraffin sections fixed in Bouin-Hollande solution were immunostained with rabbit antiserum against VIP. With the electron microscope we were able to identify nerves in the glandular stroma of all the glands examined with the exception of goblet cells. Intraepithelial single axons were only seen in the parenchyma of Wolfring glands. The morphological findings corresponded with the immunological finding of VIP-positive, nerve-like structures in the same locations, with the exception of lanugo hair follicle glands, and goblet cells. Our findings indicate that the glands of the eyelids and main lacrimal gland represent a functional unit with VIP as a possible common stimulating factor.

Exocrine Glands↗

Acute corneal necrosis after excimer laser keratectomy for hyperopia.

OBJECTIVE: To describe a new, rare clinical complication after routine excimer laser photorefractive keratectomy to correct hyperopia. DESIGN: Case report with clinicopathologic correlation. MAIN OUTCOME MEASURES: Four weeks after treatment with excimer laser, a perforating keratoplasty was performed for persistent corneal opacities. The corneal button was examined using light and electron microscopy. Special immunohistochemical stains were used to detect apoptosis. RESULTS: The patient developed corneal opacities, endothelial precipitates, and a fibrinous exudate in the anterior chamber after the laser treatment. The changes did not respond to therapy directed against bacteria, fungi, and Acanthamoeba. All examinations and special stains were negative for micro-organisms. By light microscopy, an anterior zone of corneal necrosis was present with a moderate amount of acute inflammatory cells. At the interface between necrotic and viable corneal stroma, keratocytes with typical features of apoptosis were detected by immunohistochemistry and electron microscopy. CONCLUSION: This is the first full histopathologic report of a case of acute corneal necrosis with signs of apoptosis after excimer laser therapy of the cornea. Surgeons should be aware of this rare but potentially severe complication.

Acute Disease↗

Long-term ultrastructural changes in human corneas after tattooing with non-metallic substances.

AIM: To investigate the ultrastructural appearance and the deposition pattern of dye particles in long term non-metallic corneal tattooing. METHODS: Two tattooed human corneas were obtained by keratoplasty. One corneal button was fixed in Karnovsky's solution and the other in Trumps' solution. Both corneas were divided and processed for conventional light (LM) and transmission electron microscopy (TEM). Five additional formalin fixed corneas with tattoos were retrieved from paraffin for TEM. The time between tattoo and removal of the corneal button/enucleation ranged from 7 to 61 years. All seven corneas were examined using a Jeol JCXA733 microprobe for wave length dispersive analysis in order to exclude any presence of metallic salts in the tattooed area. RESULTS: Histologically, clumps of brown-blackish granules were present mainly in the mid stroma, but also in anterior and partially in the posterior half of the stroma. On TEM, numerous round and oval electron dense particles were seen in the cytoplasm of keratocytes arranged as clusters or large islands. The larger particles appeared black, while the smaller particles were grey. In well fixed tissue a unit membrane was observed around these clusters. No granules were detected in the extracellular matrix. CONCLUSIONS: Keratocytes can actively ingest and retain tattooing particles of non-metallic dyes within their cell membrane for very long periods of time.

Adolescent↗

Hydroxyapatite orbital implant. A light- and electron-microscopic and immunohistochemical study.

OBJECTIVE: This study aimed to characterize the constituents of the ingrown host tissue in explanted orbital coral spheres by means of light electron microscopy and immunohistochemistry and to compare findings at different intervals after implantation. DESIGN AND METHODS: A microscopic retrospective laboratory study of three explanted coral spheres from three different patients was undertaken. The first specimen was an infected hydroxyapatite orbital implant, which was removed after 18.5 months and processed for standard light microscopy and transmission electron microscopy. Immunohistochemistry was performed using vimentin, actin, CD-68, and CD-34 marker. Two other explanted spheres (after 3 weeks and 2.5 months) were examined by light microscopy only, because no fibrovascular ingrowth could be detected. RESULTS: On light microscopy, the pores of the first coral were filled entirely with the fibrovascular tissue. A basophilic line reminiscent of calcium deposition was seen along hydroxyapatite spicules, where it often was associated with newly formed bony lamellae. Similar lines were seen partially in the 2.5-month specimen but not in the 3-week specimen. Immunohistochemistry of the first specimen enhanced visualization of the capillary network when the CD-34 marker was used. The fibrovascular tissue was uniformly vimentin-positive and partially actin-positive. On transmission electron microscopy, typical fibroblasts and myofibroblasts were observed. The latter were separated by thin sheets of banded collagen and formed pseudoepithelial structures. Proliferating vessels often showed high endothelial cells. CONCLUSIONS: The fibrovascular ingrowth contains not only vimentin-positive regular fibroblasts but also actin-positive myofibroblasts. Interaction between the invading tissue and hydroxyapatite structures leads to long-term ossification.

Actins↗