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Biomedical subjects

J Vogel

Publications and source records attributed to J Vogel.

At least 55 records · Page 3Linked to original sources
PubMed1999

Targetability of novel immunoliposomes prepared by a new antibody conjugation technique.

In order to develop long-circulating immunoliposomes (IL), which combine sterical stabilization with a superior targetability, we have introduced a new methodology for attaching monoclonal antibodies directly onto the distal ends of liposome-grafted polyethylene glycol (PEG) chains. Therefore, we have synthesized a new PEG-PE derivative, which had been endgroup-functionalized with cyanuric chloride. Antibodies can simply be coupled to this membrane anchor in mild basic conditions (pH 8.8) without the need for previous antibody derivatizations. The coupling results have been determined with consideration to various liposome parameters and have been compared to several established antibody coupling procedures, where antibodies had been linked directly to the liposome surface in the presence of PEG (conventional IL). To investigate the targetability of the resulting new IL, anti E-selectin mAb have been coupled and the degree of binding selectin-containing cells has been analyzed. The terminal coupled antibodies show a 1.8-fold higher degree of in vitro cell binding compared to conventional IL, which has been attributed to the antibody position being more easy accessible at the PEG termini. Furthermore, we have illustrated the liposome surface topology and the coupled antibodies by atomic force microscopy, which for such fluid IL has been used first. These images have finely corresponded to the cell binding results, and have been discussed in terms of antibody position and flexibility at the liposome surface.

Animals↗
PubMed1999

Non-operative management of arterial liver hemorrhages.

A retrospective evaluation of embolotherapy in patients with arterial liver hemorrhages was carried out. Twenty-six patients, ranging in age from 10 days to 77 years with active arterial liver hemorrhages, underwent non-surgical embolotherapy. Bleeding was attributed to trauma (n = 21), tumor (n = 3), pancreatitis (n = 1), or unknown cause (n = 1). Twenty-nine embolizations were performed via a transfemoral (n = 26) or biliary (n = 2) approach. One bare Wallstent was placed into the common hepatic artery via to an axillary route to cover a false aneurysm due to pancreatitis. Treatment was controlled in 4 patients by cholangioscopy (n = 2) or by intravascular ultrasound (n = 2). Prior surgery had failed in 3 patients. Intervention controlled the hemorrhage in 24 of 26 (92%) patients within 24 h. Embolotherapy failed in 1 patient with pancreatic carcinoma and occlusion of the portal vein. In 1 patient with an aneurysm of the hepatic artery treated by Wallstent insertion, total occlusion was not achieved in the following days, as demonstrated by CT and angiography. However, colour Doppler flow examination showed no flow in the aneurysm 6 months later. Complications were one liver abscess, treated successfully by percutaneous drainage for 10 days, and one gallbladder necrosis after superselective embolization of the cystic artery. Embolization is a effective tool with a low complication rate in the treatment of liver artery hemorrhage, even in patients in whom surgery has failed.

Adolescent↗
PubMed1999

NF-kappaB is activated and promotes cell death in focal cerebral ischemia.

The transcription factor NF-kappaB is a regulator of cell death or survival. To investigate the role of NF-kappaB in neuronal cell death, we studied its activation in a rodent model of stroke. In the ischemic hemisphere, NF-kappaB was activated, as determined by increased expression of an NF-kappaB-driven reporter transgene, nuclear translocation of NF-kappaB in neurons and enhanced DNA binding of NF-kappaB subunits RelA and p50. In p50 knockout mice, ischemic damage was significantly reduced. This indicates a cell death-promoting role of NF-kappaB in focal ischemia. NF-kappaB may provide a new pharmacological target in neurologic disease.

Animals↗
PubMed1999

Evolution of microcirculatory disturbances after permanent middle cerebral artery occlusion in rats.

Nonischemic brain capillaries show a continuous and heterogeneous plasma perfusion. In the current study, plasma perfusion was investigated in rats during 2 to 168 hours of permanent middle cerebral artery occlusion. Perfused capillaries were detected in brain cryosections by fluorescein isothiocyanate (FITC) dextran after 10 minutes of circulation time. Heterogeneity of capillary perfusion was identified by Evans blue (EB), which circulated for 3 seconds. In this setting, the heterogeneity of intracapillary EB concentrations reflects heterogeneities in capillary flow velocities. The CBF was quantified by simultaneous iodo[14C]antipyrine autoradiography. When moving from normal flow to low-flow areas in the ischemic hemisphere, three states of capillary filling could be distinguished: state 1--fast perfusion, filling by FITC dextran and EB (CBF 0.33 mL x g(-1) x min(-1)); state 2--delayed perfusion, only FITC dextran filling (CBF 0.104 mL x g(-1) x min(-1)); state 3--minimal perfusion, no dye filling (CBF 0.056 mL x g(-1) x min(-1)). In tissue of state 1 at the borderline to ischemic tissue, a higher heterogeneity of intracapillary EB concentration (85.7%) was found than in the contralateral nonischemic hemisphere (76.4%) (P < 0.05), indicating a compromised microcirculation. The adjacent ischemic areas were filled by FITC dextran (state 2) 2 to 4 hours after middle cerebral artery occlusion, indicating a maintained, although slow, perfusion at this time. Later, minimal perfused areas (state 3) progressively replaced the delayed perfused areas (state 2). This study shows, for the first time, the evolution of microvascular disturbances in relation to CBF. In the low-flow areas, an early residual plasma perfusion is later followed by a lack of perfusion or minimal perfusion. In areas of higher, although reduced flow at the border between normal and ischemic tissue, an extreme capillary perfusion heterogeneity indicates permanent microcirculatory abnormalities.

Animals↗
PubMed1999

Cytomegalovirus-induced transendothelial cell migration. a closer look at intercellular communication mechanisms.

A variety of cells such as leukocytes and tumor cells may adhere to endothelial cells and subsequently transmigrate into the solid tissue by involving specific intercellular molecular pathways. One important prerequisite for transendothelial migration is the loosening of endothelial cell-to-cell contact sites, which can be triggered by extravasating cells. Cytomegalovirus (CMV) has obviously evolved the ability not only to influence host cells floating in the blood stream to adhere to endothelial cells, but also to induce the formation of intercellular gaps within the endothelium, resulting in transendothelial migration. These features allow the virus to disseminate and evade the immune system. In coculture experiments with human endothelial monolayers and human CMV (HCMV)-infected neuroblastoma cells or leukocytes, changes in the integrity of the monolayer were observed and further analyzed on the molecular level. For example, HCMV may activate the integrin beta1alpha5 (VLA-5) that triggers adhesion to endothelial cells with subsequent focal disruption of endothelial cell-to-cell connections. It is hypothesized that a Ca(2+)-independent pathway following VLA-5 binding disconnects the cadherin-catenin-actin complex within the endothelial cells. The loss of cadherin function causes the loss of contact to the neighboring endothelial cells and thus could represent an important mechanism in HCMV-induced cellular transendothelial migration and disruption of the endothelial integrity.

Cell Adhesion↗
PubMed1999

Early delineation of ischemic tissue in rat brain cryosections by high-contrast staining.

BACKGROUND AND PURPOSE: After short periods of ischemia, commonly used staining methods yield only moderate differences in optical contrast between normal and damaged brain tissue when gray-scale images are used for computer-assisted image analysis. We describe a high-contrast silver infarct staining (SIS) method that allows an early delineation of ischemic tissue as soon as 2 hours after middle cerebral artery occlusion (MCAO) in rat brain cryosections. METHODS: Rats were subjected to permanent MCAO for 2, 4, 6, and 48 hours. The optical densities were quantified in nonischemic white and gray matter and in damaged tissue from gray-scale images of serial sections with the use of a video camera-based image analyzing system. SIS, hematoxylin-eosin, Nissl, and nitroblue tetrazolium stainings were performed in cryosections, and 2,3, 5-triphenyltetrazolium hydrochloride (TTC) staining was performed in unfrozen vibratome sections. In addition, the range of reduced cerebral blood flow (CBF) in areas demarcated by SIS was determined in iodo[14C]antipyrine autoradiograms of adjacent cryosections. RESULTS: At all times after MCAO, only SIS showed significantly (P<0.01) lower optical densities in damaged than in normal brain tissue for both white and gray matter. TTC staining was as effective as SIS 6 and 48 hours after MCAO. The tightest correlation between areas of reduced SIS and of reduced CBF was found at a mean ischemic CBF of 22.3 mL/100 g per minute. This corresponds to a CBF range of 0 to 44 mL/100 g per minute in areas of reduced SIS. CONCLUSIONS: In contrast to other staining methods, SIS allows a reliable delineation of ischemic brain tissue (core plus penumbra) from nonischemic white and gray matter of rat brain cryosections as soon as 2 hours after MCAO.

Animals↗
PubMed1999

Detection of liver metastases from pancreatic cancer using FDG PET.

UNLABELLED: We evaluated the potential of the glucose analog [18F]fluorodeoxyglucose (FDG) as a PET tracer for the hepatic staging in 168 patients designated for resective pancreatic surgery. METHODS: Metastatic liver disease was confirmed or excluded during surgery or with CT follow-up for at least 6 mo. Proven metastases were then retrospectively identified on preoperative CT (gold standard). Hepatic PET scans of all patients were interpreted blindly. Any focal FDG uptake was considered malignant. Both proven hepatic metastases and suspicious hepatic PET lesions were then compared, lesion by lesion, with CT. Standardized uptake values (SUV) and tumor-to-liver ratios (T/L) were determined for the most intense lesion of each patient. RESULTS: Sensitivity of FDG PET was 68% (15 of 22 patients). The lesion detection rate was 97% (28 of 29 metastases) for lesions >1 cm and 43% (16 of 37 metastases) for lesions < or = 1 cm. Specificity was 95% (138 of 146 patients). Six of eight patients with false-positive results had marked intrahepatic cholestasis (versus 3 of 15 patients with true-positive lesions), one had an infrahepatic abscess and one had a right basal pulmonary metastasis. The SUV and T/L were 4.6+/-1.4 and 2.3+/-1.1, respectively, for malignant lesions and 4.1+/-1.5 and 1.9+/-0.3, respectively, for false-positive lesions and therefore are of limited value. CONCLUSION: FDG PET provides reliable hepatic staging for lesions >1 cm. False-positive results are associated with the presence of marked intrahepatic cholestasis. For lesions < or = 1 cm, FDG PET can define malignancy in 43% of suspicious CT lesions in the absence of dilated bile ducts.

Aged↗
PubMed1999

Fibrinolysis and aspiration of experimental intracerebral hematoma reduces the volume of ischemic brain in rats.

The hypothesis was tested in rats that brain ischemia by an intracerebral hematoma can be ameliorated by fibrinolysis and aspiration of the hematoma. Intraparenchymal blood clots were generated by the injection of 50 microliters of autologous blood into the right caudate nucleus in two portions seven minutes apart. Thirty or 120 min later 12 microliters recombinant tissue plasminogen activator (rtPA) or 0.9% NaCl were injected and after 30 min the resolved hematoma was aspirated. Six hours later cerebral blood flow (CBF) was determined by 14C-iodoantipyrine autoradiography. Tissue volumes of CBF < 10 ml 100 g-1 min-1 and CBF < 30 ml g-1 min-1 were determined. Clot and lesion volume were quantified histologically from serial sections stained for succinate-dehydrogenase (SDH) activity. In rtPA-treated rats the major part of the hematoma could be evacuated 30 min as well as 120 min after production of the clot. The volume of ischemic brain (CBF < 10) was significantly reduced (p < 0.05) in the rtPA group compared to saline-treated and control groups irrespective of the time of treatment. In contrast, no difference was found between the control group and the experimental groups when the volumes of brain tissue surrounding the lesion were compared which had values of CBF < 30 ml 100 g-1 min-1. In a rat model of intracerebral hemorrhage, treatment by local fibrinolysis followed by aspiration of the hematoma is effective in reducing the volume of ischemic brain tissue and of the remaining clot volume.

Animals↗
PubMed1998

Capillary patency after transient middle cerebral artery occlusion of 2 h duration.

Reperfusion after transient focal ischemia of 2 h duration is followed by secondary bioenergetic failure after 4 h of reperfusion. The objective of the present study was to explore whether or not this secondary deterioration is due to secondary microcirculatory compromise. Normal fasted rats were subjected to 2 h of MCA occlusion and allowed reperfusion for 2, 4, 6 and 8 h. At predetermined reperfusion times, rats were injected with Evans blue and decapitated. Capillary patency was determined using a fluorescent double-staining technique. No capillary perfusion deficits were detected in the ischemic neocortical penumbra, neocortical focus or striatal focus. We concluded that the secondary deterioration of bioenergetic state is not due to microcirculatory compromise. Since hyperglycemic animals show pan-necrotic lesions, a hyperglycemic group was added at 8 h of reperfusion to test if the adverse effect of hyperglycemia on ischemic damage is related to capillary compromise. The results showed that, in hyperglycemic rats, capillary perfusion in the striatal focus was compromised after 8 h of recirculation following 2 h of MCA occlusion. It is concluded that when normoglycemic rats are subjected to 2 h of MCA occlusion, capillary patency is not affected during the first 4-6 h of reflow. At 8 h of reflow, though, particularly in hyperglycemic rats, microcirculation is compromised in the caudoputamenal focus, probably reflecting infarction.

Animals↗
PubMed1998

Hyperglycemia-exaggerated ischemic brain damage following 30 min of middle cerebral artery occlusion is not due to capillary obstruction.

Transient focal ischemia of brief duration (15-30 min) gives rise to brain damage. In normoglycemic animals this damage usually consists of selective neuronal necrosis (SNN), and is largely confined to the lateral caudoputamen. In hyperglycemic subjects damage occurs more rapidly, involves also neocortical areas, and is often of the pan-necrotic type ('infarction'). Since experiments on forebrain ischemia of 30 min duration suggest that microcirculatory compromise develops during recirculation, we studied whether focal ischemia of the same duration, followed by reperfusion for 1, 2 or 4 h, leads to microcirculatory dysfunction. To test this possibility, we fixed the tissue by perfusion and counted the number of formed elements (leukocytes, macrophages and erythrocytes) in capillaries and postcapillary venules. Furthermore, capillary patency was evaluated following in vivo injection of Evan's blue. Histopathological examination of tissue fixed by perfusion after 1, 2 and 4 h of recirculation showed an increasing density of SNN in the caudoputamen of normoglycemic animals. Hyperglycemic, but not normoglycemic, animals showed pan-necrotic lesions ('infarction') after 4 h of recirculation. As a result, the total volume of tissue damage (SNN plus infarction) was larger in hyper- than in normoglycemic animals at 2 and 4 h of recirculation. In addition, hyperglycemic animals showed involvement of neocortex which increased with the time of reperfusion. In the ischemic hemisphere, between 5 and 10% of counted capillaries contained formed elements. However, since hyperglycemic animals contained an equal (or smaller) amount of cells the results did not suggest that capillary 'plugging' could explain the aggravated damage. Moreover, both normo- and hyperglycemic animals showed close to 100% capillary patency. The results thus fail to support the notion that the aggravation of focal ischemic damage by hyperglycemia is due to obstruction of microvessel by swelling or leukocyte adherence.

Animals↗
PubMed1998

The role of glycolipids in mediating cell adhesion: a flow chamber study.

Selectins constitute a family of proteins that mediate leukocyte tethering and rolling along the vascular endothelium by recognizing various carbohydrate ligands in response to inflammation. To test the hypothesis that multivalent binding of selectins to their ligands is the molecular basis for achieving sufficient binding forces, we have performed this flow chamber study. Selectin-containing Chinese hamster ovarial cells (CHO-E) bind and roll along a support-fixed phospholipid membrane containing a defined concentration of a synthetic Sialyl Lewisx (sLex) glycolipid ligand. Ligands are either homogeneously distributed, or arranged in defined lateral clusters, as illustrated here for the first time. The lateral glycolipid clusters which appear as recognition motifs are essential for mediating cell rolling. Furthermore, the transition from firm cell adhesion to cell rolling depends on the site density of ligands. Rolling velocity shows little dependence on shear forces within a broad range. As we found out that cells do not roll along the model membranes with homogeneous ligand distribution, our results therefore support the hypothesis of multivalent binding events. Since these investigations suggest that lipid-anchored sLex, functionally embedded in a lipid matrix, can mediate cell rolling, this study demonstrates the relationship between dynamic glycolipid binding to selectins with the hypothesis of multivalency of binding for the first time.

Animals↗
PubMed1998

Preischemic hyperglycemia leads to rapidly developing brain damage with no change in capillary patency.

The present experiments were undertaken to explore whether exaggeration of ischemic brain damage by preischemic hyperglycemia is due to lack of capillary patency in the postischemic period. Normo- and hyperglycemic rats were exposed to 10 min of forebrain ischemia. Histopathological changes were evaluated after 6 and 16-18 h of recovery by light microscopy, and capillary patency was assessed at the same time points by a double-staining technique, depicting perfused and morphologically identifiable capillaries. The results demonstrate that some neuronal damage was detectable after 6 h of recirculation which was aggravated after 16-18 h of recirculation in hyperglycemic rats. In contrast, the degree of capillary patency was similar in normo- and hyperglycemic rats. In both groups the perfusion marker, Evans blue, perfused about 95% of all capillaries when injected 10 s before decapitation. Since preischemic hyperglycemia exaggerates brain damage without cessation of capillary perfusion the primary targets of hyperglycemic brain damage may not be capillaries but neurons or glial cells.

Animals↗
PubMed1998

Selectins as new targets for immunoliposome-mediated drug delivery. A potential way of anti-inflammatory therapy.

Endothelial cell adhesion molecules, which are expressed in response to inflammatory signals to mediate recruitment of leukocytes to sites of inflammation, appear to be excellent targets for drug delivery systems to open new perspectives of antiinflammatory therapies. In this study we describe the preparation and characterization of antibody-coupled liposomes (immunoliposomes) as directed against endothelial (E)-selectins. We have examined the factors affecting the covalent coupling of antibodies to the membrane anchor N-glutaryl-phosphatidylethanolamine via amide bound and have compared them to other coupling procedures. The target sensitivity has been demonstrated in a cell-containing in-vitro model, where liposome binding to selectins under either static, or simulated blood flow conditions was illustrated by using fluorescence microscopical means. It could be shown that even under shear force conditions liposomes selectively accumulate at selectin-containing cells when a specific lipid composition and a certain balance in the lipid/antibody ratio was maintained. Furthermore, the need for polyethylene glycol-derived lipids to sterically stabilize the liposomes for preventing unspecific liposome attachment to cells has been demonstrated.

Animals↗
PubMed1998

Measurements of chain length distributions in calcium phosphate glasses using 2D 31P double quantum NMR.

31P double quantum (DQ) NMR is used for improved studies of the structure of phosphate glasses. The common Qn notation of the network forming tetrahedral phosphate units with n bridging oxygen atoms is extended to Qn,jkl indicating the Q type of the bonded adjacent units by the additional superscripts j,k and l. It will be shown that: (i) Q1 units have different isotropic chemical shifts depending on whether they form diphosphate anions in the amorphous state (Q1,1) or end groups of chains (Q1,2) and (ii) that even Q2 units possess different isotropic chemical shifts such that in the DQ dimension Q2,11 can be distinguished from Q2,12 and middle groups of longer chains or rings (Q2,22). Based on this result the opportunity for measuring chain length distributions in the amorphous state is discussed and first results for binary calcium phosphate glasses are presented. The advantages and also the limitations of this novel approach will be discussed.

Calcium Phosphates↗
PubMed1998

[Measurement of migration of acetabular components in cementless hip replacement].

PURPOSE: Migration measurements of acetabular components using a special computer aided method (EBRA = abbreviation for the German term "Ein-Bild-Röntgenanalyse") were performed to evaluate early results of the implants and predict aseptic loosening. METHODS: Standard ap-radiographs of the pelvis were marked, specific points were digitised. Simulating the spatial situation the programme computes longitudinal and vertical migration of the cup. 74 acetabular components in 71 patients could be studied by migration measurements. RESULTS: 14 patients showed migration of more than 1 mm, which is the confidence limit of this method. Each of these patients showed diverse reasons for the migration, i.e. osteoporosis of the acetabular bone stock or problems concerning the surgical technique which means malposition of the cup or insufficient reaming of the bone. There were some patients with severe congenital dysplasia of the hip and in some cases the inclination angle of the cup was too great. CONCLUSION: The technique applied for measuring migration of acetabular components can be useful for evaluating early instability of the implant and can be helpful in detecting problems concerning the surgical technique.

Acetabulum↗
PubMed1998

Local fibrinolysis and aspiration of intracerebral hematomas in rats. An experimental study using MR monitoring.

Serial magnetic resonance (MR) imaging has not yet been validated in the therapy of experimental intracerebral hematomas in a rat model. It is possible to test the effect of local fibrinolysis and aspiration on the clot volume using serial magnetic resonance imaging and different MR-sequences. Experiments were carried out in 22 male Sprague-Dawley rats. Intracerebral hematoma was produced by injection of fresh autologous blood into the caudate nucleus using a double injection technique. Thirty minutes later 10 rats were treated by injecting 12 microliters of recombinant tissue plasminogen activator. MR-imaging was performed immediately after generation of the hematoma and after clot lysis. The clot volume measured in the magnetic resonance images was compared with that obtained in stained histological serial sections at the end of the experiment. Serial MR scanning demonstrated a significant reduction (p < 0.01) of hematoma volume after fibrinolysis followed by aspiration of the blood clot. The best correlation between MR- and histological volumetry was found on RF-spoiled FLASH 2D-images. This study documents the efficacy of MRI in detecting and delineating the size of acute intracerebral hematomas and its time course. Local fibrinolysis and aspiration can be simulated in an experimental rat model.

Animals↗
PubMed1997

Splicing and intron-internal RNA editing of trnK-matK transcripts in barley plastids: support for MatK as an essential splice factor.

Group II introns frequently require assistance by specific factors, maturases, for folding and effective splicing in vivo. The only putative maturase of higher plant chloroplasts is encoded by matK, located in the intron of trnK. We show that in barley matK transcripts are modified at a first codon base by C-to-U RNA editing. The resulting H --> Y substitution restores a sequence motif that is present in maturases of yeast and plant mitochondria and of Lactococcus ltrA and that is positioned within the X domain. Processing of trnK-matK transcripts was further investigated in plastids lacking functional ribosomes due to a mutation. Absence of the intron-encoded matK gene product in these plastids is correlated with the accumulation of precursor transcripts for tRNALys(UUU)-matK, processed to different degrees, and by the lack of mature and spliced tRNA molecules. These results suggest an essential role of MatK for splicing of its own transcript in vivo. Processing of the 5' end of trnK exon 1 was found to proceed efficiently also in the mutant plastids although the two tRNA exons were separated by the 2481 nt intron. Consequently, presence of the intron does not interfere with the formation of mature 5' termini.

Amino Acid Sequence↗